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Engineers designing electronic systems likely have experience working with field-programmable gate arrays (FPGAs), programmable hardware chips that can be overlaid with logic to create electronic components—and even entire systems. These chips provide the ability to quickly configure and customize electronic devices after manufacturing, in the field.
In many cases, FPGAs and similar logic chips are also used to prototype complete electronic systems that are used in system-on-a-chip designs that deliver full processing and input-output functionality for applications like mobile phones and automotive electronics. These prototypes are then cast into application-specific integrated circuits (ASICs) for performance and manufacturability, and used in complex hardware-software systems.
But engineers can have a difficult time working with FPGAs, and deeply embedded systems in general. FPGAs and other logic devices have hundreds of thousands, or even as many as tens of millions of logic gates, and programming them using traditional hardware definition languages can be a challenge. Most designers prefer working with pre-defined logic components, referred to as intellectual property, or IP. These are hardware description language (HDL) programs that define a particular function, such as a processor, I/O interface like USB, or even graphics.
Further, these systems are almost always customized with software. An automotive braking system, for example, uses a processor, I/O for data inputs and instructions, and storage. But it is also coded with explicit instructions on how to interpret and respond to data inputs, and how to handle data and processing errors.
This embedded software is difficult to develop while the hardware design and implementation are still in flux. Interfaces, timing and even performance aren t usually fully defined until the hardware system is designed and the components selected. That puts software development behind the hardware, typically at the very end of the project. Putting software last often has undesirable consequences, such as poor performance, bad design, incomplete or missing features, and poor testing.
Building Virtual Prototypes in Logic Devices
In prototyping, electronic designers partition the FPGA logic chips and position the IP logic onto the chip, then design interfaces so that they work together in providing the needed functionality. While this may not be the final design, work on the software can begin as soon as there is an initial prototype of the hardware on the FPGA. The resulting circuit is then tested for functionality in the larger system, or within a testing harness.
Once hardware is finalized, it can be placed on the FPGA and the software can be completed. The device can be used as-is, if the end product is custom or for a small manufacturing run. For large runs, the FPGA can be produced as an ASIC or a system on a chip (SOC), and the costs spread over a larger product base.
A typical design flow for embedded software optimization of electronic hardware-software systems, using TLM Central system models and the Virtual Development Kit.
This prototyping process sounds straightforward, but it often isn t. IP blocks can be difficult to work with, especially if multiple blocks are being used on a single chip. Often, the position, layout and interfaces of these blocks can influence both the capacity and performance of the resulting circuit. Sloppy design can easily result in not meeting product specs or performance. As embedded electronic systems become more complex, the costs of bad design and placement become more significant.
Vendors such as Xilinx and Altera make the FPGAs and other programmable logic devices that are used in prototyping these embedded software systems. The hardware description languages are standard across the industry. The competitive advantage in creating prototypes comes in the placement and mapping software that determines the optimal places for IP blocks on a chip. The best approaches to prototyping can save significant time and chip expenses over manual methods.
Hardware-Software Co-development
An initiative by the programmable logic industry is making it possible to do more embedded software development as a part of the hardware prototyping process. This initiative, called TLM Central, consists of a portal, a set of high-level descriptions and models of standard hardware components in the SystemC language, and comprehensive information on how to create and manage virtual prototypes.
These virtual prototyping systems are integrated with an intelligent prototyping software environment that enables faster partitioning, and automates the creation and debugging of prototypes for a range of designs—from individual IP blocks and processor sub-systems to complete SOC, easing the path from concept to operational prototype. Engineers can use the on-chip virtual prototype as a platform for loading an operating system, coding control and management software unique to the product.
The important part of the process is that the FPGA-based prototype is used for software development while the hardware is still being defined. The TLM Central solution includes a set of freely available software development components integrated into the engineer s software development environment.
According to Tom De Schutter, senior product marketing manager, System Level Solutions at founding member Synopsys, the more engineers that use TLM Central, the more valuable it becomes: “With over 1,000 hardware component models, engineers can build virtual simulations of a wide variety of hardware-based embedded systems in many different industries.”
The accompanying software environment, called a Virtual Development Kit, incorporates popular software debuggers, system control mechanisms, and debugging information. To work with a virtual prototype, engineers install and start using—there s no need to wait for a hardware prototype to develop software.
Possibly the biggest advantage in this approach is a reduction of project risk. Any major issues with the software can be worked out more easily, either through redesigning interfaces or substituting other hardware components.
Engineers engaged in complex hardware-software system design in mobile communication, automotive, aerospace and similar industries are often faced with a rush to complete software development and integration once the hardware is in place. Using a virtual prototype of an FPGA and TLM Central-based system models, designers can begin work on the software systems before the hardware components are fully defined. Once final silicon is ready, either on an FPGA or ASIC, product designers can finalize the software much more quickly. Most important, there is no need to wait until final silicon to begin software development.
It s not a panacea, of course. Any software written before final silicon will have to be tested—and likely undergo some changes once it is able to run on actual hardware. And while the two can be developed more or less in parallel, the software will always take longer. But using a virtual prototype can reduce risk at the same time it accelerates time to market for an optimized design. DE
Contributing Editor Peter Varhol covers the HPC and IT beat for DE. His expertise is software development, math systems, and systems management. You can reach him at [email protected].
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The film adaptation of Markus Zusak’s 2005 bestselling novel was directed by Brian Percival and released in November 2013. The Book Thief is the story of a young Liesel Meminger growing up in Nazi Germany. We are introduced to her by Death, who is unusually intrigued by Liesel’s quiet perseverance. He first encounters Liesel when her younger brother Werner dies in their mother’s arms. Liesel steals her first book at Werner’s funeral before being dropped off at the home of her new foster parents, Hans and Rosa Hubermann. With Hans’ help, Liesel learns to read, thus developing a voracious appetite for words and pilfering books at every opportunity. When the Hubermanns shelter a Jewish man named Max, Liesel learns the dire consequences of war and the power of storytelling.
After watching the film, I decided that while I wasn’t entirely satisfied with the film as an adaptation, it was a perfectly enjoyable movie, mostly due to the fantastic casting and beautiful cinematography.
Thirteen-year-old Sophie Nélisse is a luminous Liesel. The scenes between Liesel and her best friend Rudy, played by an impish Nico Liersch, are full of joyful innocence and show how children can be loyal, perceptive, and brave. Nélisse is wonderfully supported by Geoffrey Rush as Hans and Emily Watson as Rosa. I found Watson’s portrayal of Rosa to be particularly endearing. Her Rosa, while just as gruff and harsh as she is in the book, shows convincing glimpses of levity, maternal love, and compassion. Liesel and the audience are immediately drawn to Hans’ warmth and humor, but it was Watson’s Rosa who gave me a newfound appreciation for her prickly character.
It should be noted that faithfully adapting a 550-page novel into a movie can be a difficult undertaking. There is no way every memorable scene from the novel can fit into the allotted timeframe. In 131 minutes, the film attempts to cover most of the key storylines from the novel but doesn’t quite manage to give them all sufficient development.
Liesel, Hans, Rosa, and Max come together as a makeshift family bonded by compassion and hope. Death’s voice-overs are used perfunctorily to move the story along and neatly wrap up the film. One noticeable change from the novel is that some of the interactions that made the relationship between Max (Ben Schetzer) and Liesel compelling are missing or glossed over. In the film, the audience is given very little background information about Max. The audience learns that his father saved Hans’ life during World War I and that he was forced to leave his mother behind when fleeing his home during a Nazi invasion. There is no mention of his history as a fist fighter, or how he passes the time hiding in the Hubermann’s basement by fantasizing about fighting Hitler in a boxing ring. The most glaring omission was the handwritten book Max presents to Liesel after he wakes up from a life-threatening fever. In the novel, the thirteen pages depicting his life story demonstrate the deep friendship and remarkable bond Liesel and Max share over their love for words and creative storytelling. Personally, it was one of the most poignant passages in the book. While there is an obvious connection between the two in the film, it was disappointing to see so little development of an important character.
Also, for a movie set in a perilous time in history, the dangers are often implied, rather than shown. It was perhaps assumed that the audience is already fully aware of Hitler and the Nazi party’s historical significance. The scenes of children singing Nazi propaganda and Liesel desperately searching for Max in a crowd of Jewish prisoners are dramatic but restrained enough to be suitable for a young audience.
Overall, the film is heartwarming and enjoyable. Liesel’s discovery of the immense pleasure one can receive from reading and creating stories, even during times of fear and loss, will hopefully inspire many young readers to pick up or revisit the original source material.
If you enjoyed the film adaptation of The Book Thief, be sure to vote in our 2014 Literary Fauxscars. Sophie Nélisse is nominated for “Best Character Portrayal by an Actress,” and the film is up for our “Best ‘Young Adult’ Adaptation” award.
For more on The Book Thief, read a review of the novel from our archives. | https://www.literarytraveler.com/books/the-book-thief-and-the-power-of-storytelling/ |
Sunday, March 8, 2009
It is so nice hearing from you in this spirit. I would further this discussion through writing even more. But before I begin, I would like first of all to establish certain preliminaries having nothing to do in person. I would reply to these questions in the same spirit even if you are a complete stranger. However, since I have known you only for some time, it would make more sense to me and you both that we understand what as individuals, leaving aside our personal or social network in terms of friendship or relationship by birth or marriage, ought to get from our lives. It may necessarily vary from person to person for their personal location on time line and in space, but one thing is very much sure every soul in this world undergoes these experiences and tries to find the best for their happiness, which is obviously a relative and subjective term, though, of course, it is a state of mind.
The multi-dollar question for every one is: what helps one attain this state of mind? And why does not every being (particularly human beings) attain this state of mind, if it is that relative and subjective? What forces an individual to stop attain this state of mind? And there are other related queries and questions to be answered. Now that the larger questions are clear to me, I might address them properly from my point of view and also reflect on how come I gathered courage to understand happiness, if at all, I have come across ETERNAL BLISS in this physical world or the mental world. Also who am I to question an individual's happiness (such that of yours) and in what spirit, if at all I do so, I would do it. Now listen carefully:
Primarily, please do not take this issue as personal first and then, don't take it to the matter of personal ego. Secondly, you must trust this unknown soul in its entirety, inasmuch as I am not doing it for any personal reasons but for a common cause from this point onwards. If it were so personal and friendly alone, I would not have bothered talking this much because this is quite sensitive issue and I have no right to talk in this spirit about any person or group in any capacity so far he or she is not a matter of public debate.
In sum and substance, I do not have any right to negatively critique any person (or group) about his or her personal take of the life and its varied aspects. Finally, I am doing so as to lift you up from where you are and the kind of pain you are either scared of or you are undergoing. I am not talking because I am a friendly or otherwise but I am talking because I am not a single soul to undergo such a pain if there is any, which I observed in you (once again I apologize for saying so, otherwise I do not have any right to say so). But now that I made things clear, I would start speaking in the light of questions posed earlier. Before I come back to the questions raised by me in the earlier paragraph, I would like to answer your questions:How can you [Vishwajeet, I would refer to me as him henceforth] know [An INDIVIDUAL] so much in such a short time? First of all, what is knowing a person i.e. when he says. "I know you"; what it is that he knows about a person.
So what is this knowledge that he claims to have about An INDIVIDUAL and how much reliable his knowledge is there about him/her. In other words, what makes a human being an individual - distinct from not only from other animals; but unique in all senses from all other members of the same species and of the other member of a community in this world of 21st century. It also involves questions about personal beliefs, non-beliefs, fear/s, his or her location on the time line and in the space. What makes these things in a person? Does a social set up i.e. socialization or education or exposure to / interaction with the wider variety of communities thriving in all quarters of the world or individual's short or long term goals or all of them make one an individual? He is sure that An INDIVIDUAL would agree with him that it is every ounce of time and interaction with this world, may be, with a person or object or an event contributes significantly in making intellect of a man or a woman.
And finally, an individual (confined to human world only at this stage) does not have only sense of 'individual being' but also a 'social being'. This also suggests that knowing of a person does not involve knowing him or her in person within but also knowing of his or outer layering i.e. outer world in which he or she operates, beyond them all, he or she develops various skills to fight the odds of life, past, present and future and all. Thus, knowing a person would involve knowing him or her at all these levels. These things aside, he or she does not only become knowable but develops a keen sense of knowing others i.e. if occasion arises, he or she would be able to apply his or her knowledge to know others as and when need arises and as he or she interacts with. This happens so fast and instantly and spontaneously in a trained mind that he or she does not, at times, need to read or observe others for longer times. It is something like reading an article. Suppose An INDIVIDUAL has read fifty articles based on Second Language Acquisition. If s/he took 15 minutes per page in reading beginning articles, s/he will obviously not take more than 4-5 minutes to read the last few articles in the lot of 50. Her comprehension skills has built up so strong in the long run that s/he can read and understand it without any efforts.
Same thing happens in reading people too. Meeting people is nothing but reading a mind, though in various forms, since things are not printed in black and white, it is really little difficult but through constant practice and consistency, it is possible to read them in one go and dig into someone's mind as deep as to overwhelm people. Though it has never been his intention, it happens naturally with these people. This is how a common man or woman rises from the ground to in various terms and impress people in very many ways. For those, who have become mass-movers or group (as larger as world) leaders, only because of their being quick in grasping things with its spirits and applying them in the same spirit for future operations, it is so simple.
He claims to know An INDIVIDUAL in such a great detail that he might tell things in present and future. Though s/he might not believe it or feel reluctant to do so, not because s/he is not what s/he is, but because this mystery of an individual (what he or she is what makes him or her, what he or she is) is what gives him or her an upper hand and makes him or her distinct, thus, impressing others. S/he would be reluctant to accept only because s/he would not like this mystery to be unfolded. Because once this mystery of individuality is unfolded, he or she becomes a follower of the person who unfolds it directly or indirectly; intentionally or unintentionally. And human mind at large is so reluctant to accept this, because there lays individuality. He appreciates this approach for encouraging uniqueness and moving and living life. Otherwise, settling with other's ideas would always bring nothing but stillness and stillness is nothing but death. It is not any good time to settle with it. So, whether An INDIVIDUAL believes it and follows him or not, but for sure he understands him/her; and since he does so, he wants to draw his/her attention towards her/himself; and the weaknesses that he sees in him/her through his/her such personal interactions. He/she is voicing out his/her soulful concerns, perhaps, he/she overlooked it personally for lack of time or attention or training at one or the other level.
Therefore, with this knowledge being a stranger having nothing to do with An INDIVIDUAL, he would say that the INDIVIDUAL is a wonderful person happy with his/her life and times. But as soon as he becomes your voice, inasmuch as he understands her or say, knows her, he would say that the INDIVIDUAL is little unhappy. But first of all, he and the INDIVIDUAL must agree on the definition of HAPPINESS and its DERIVATION. Let me reveal what he understands by the much reflected term HAPPINESS. It is nothing but a state of mind of an individual putting his or her whole being (physically, mentally, psychologically, biologically, etc) into such a mode that every aspect of that being is free of worries, weariness, tension, trauma, dullness, tiredness etc that he or she radiates with energy to extend this happiness (state of mind) to others through cheerful face, active body, looking for interaction and intercourse. It is involving so much so that he or she forgets every thing rest in this world and enjoys the pleasure of current moment. He or she lives into it. This HAPPINESS is generally derived through a balancing act between his or her personal INTELLIGENT, EMOTIVE, BODILY, SPIRITUAL BEING, SEXUAL BEING, COMMUNICATIVE BEING giving rise to a healthy body, mind and soul. And the level of these factors into one's being ( i.e. IQ, EQ, BQ, SQ, XQ, CQ etc) is different in different beings, but in a happy being has it all at a balance. Any lack of balance in it brings unhappiness in the being.
Thus, he or she is observed to be unhappy or happy. A satiation of these different beings in the same individual of his or her brings him or her close to the state questioning the purpose of his or her being and the experience of ENLIGHTENMENT. There he or she acts into the direction. And once he or she knows how to strike this balance and practices it into the same direction, he or she attains ETERNAL BLISS. When he or she dies, he or she liberates from the CYCLE OF BIRTH AND DEATH i.e. LIFE. The being with a unique set of beings and balance would never be replicated and be born again in this world. But attainment of this state of being is really very difficult and not every one gets even close to it. Those who do so get IMMORTALITY.
He wishes every being in this world attain this state of being. For sure, this state of mind or being is the outcome of interaction within and balancing act within. It does never come from outside. As soon as an individual tries to get it from outside, he or she develops a tendency to depend on the outside world in one or the other sense on one or the other object or being in this world. And this is a matter of striking harmony between the two. It is next to impossible, because every being or object is unique in this world and tries to retain his or her individual nature and (say, mystery) and there acts the forces against such an act of deriving happiness from outside and not from within.
He has come across similar experiences through interacting with individualsL. The Individuals in common tend to derive happiness not from within but from outside - relaxing and depending, when there is chance to act on their own; and then, extend happiness from within. (He can be quite objective and detailed in this aspect as well, if it is required). Still he is so positive and optimistic about the individual's growth that he looks forward in an INDIVIDUAL of a tremendous change from within and derive HAPPINESS from within first and extend it when occasion demands.
It is not for his sake or benefit or for his/her interaction with one individual but all except none. There is every potential in every being of this world to do so, what is required is of little interrogation and action into that direction coming out of the confinements of fear and suspicion, feelings of solitude and fertitude. "And An INDIVIDUAL has all the potential to do so," he believes so. A short comment, people generally make, "perhaps you have overpowered me". It is human nature to overpower others in one or the other form. One should know how to resist it and act to overpower others for zest of life. An INDIVIDUAL must take his/her time to overpower others. If s/he fails, it is not other's fault who empower or overpowers him/her but of a belief system. And this system must be corrected!
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CROSS REFERENCE TO RELATED APPLICATION
FIELD OF THE INVENTION
BACKGROUND OF THE INVENTION
SUMMARY OF THE INVENTION
BENEFIT OF THE INVENTION
EMBODIMENTS OF THE INVENTION
This application claims the benefit of Chinese Patent Application No. 201810088062.0 filed on Jan. 30, 2018, incorporated herein by reference as if fully set forth herein.
The invention relates to the measurement, verification, and calibration of capacitance, including the standards of capacitance.
Capacitor standards are special-made capacitors for measurement, verification, and calibration to ensure repeatable values. In RF impedance measurement, they are the standard instruments for capacitance transferring in testing and tuning of electronic circuits, can also be used as electronic sensors, and improve the accuracy of the measurement and sensor.
The normal standard capacitors are decade capacitor including physical capacitors, rotary switches, and output ports. The existing decade capacitors have a switch with 10 output connections for 10 capacitors. It needs 20 wires to connect them together. The connection of switch and capacitors has complicated structure and long wires, which introduces large parasitic into the system. Taking an example of a decade capacitor with 10 of 100 pF capacitor, the output of capacitance are listed as input connection of 10 capacitors in Table 1.
The existing capacitor standard has none of compensation available for open and short circuits. In the pre-measurement-calibration of existing RLC bridge meter, the calibration can remove only the influence of input terminal and testing cables, but not the effect of internal connection wires and rotary switches, which will result in certain error. Especially in the case of large capacitance (low impedance), the uncertainty becomes significant. The operation procedure is complicated.
This invention provides a novel design and configuration to minimize the uncertainty and improve the accuracy by utilizing the combination of 4-value capacitors and a new decade changeover switch including short and open position to realize the decade capacitor and offer compensation for both open and short circuits inside the decade capacitor standard.
The technical proposal of the invention is as follows. In order to realize the high precision decade capacitor, the innovation design and configuration includes six special changeover switches and six sets of capacitors. Each set of capacitors has 4 physical capacitors in which one capacitor has to be single unit capacitance.
The special changeover switch can encode the selection of decade capacitance values. The changeover switch has input and output terminals. The input terminals are connected to the physical capacitors. The output terminals are connected to the output ports of the capacitor standard box. The selected capacitance is connected to the output ports of the capacitor standard box. The output capacitance can be selected in 1 unit increment from 0 to 10 times of single unit capacitance for each decade changeover switch.
For each of six sets of decade capacitors, 4 physical capacitors in special designed values are employed to encode decade capacitance values. For example, if 4 capacitor values are designed in a series of 1, 2, 2, 5 of unit capacitance (e.g. 100 nF, 10 nF, 1 nF, 100 pF, 10 pF, and 1 pF), six sets of decade capacitors will have unit capacitance values respectively in 100 nF, 10 nF, 1 nF, 100 pF, 10 pF, and 1 pF; or if 4 capacitor values are designed in a series of 1, 2, 2, 5 of unit capacitance (e.g. 100 μF, 10 μF, 1 μF, 100 nF, 10 nF, and 1 nF), six sets of decade capacitors will have unit capacitance values respectively in 100 μF, 10 μF, 1 μF, 100 nF, 10 nF, and 1 nF.
The decade capacitor box has BNC connectors in 4-terminal or 3-terminal outputs. The physical capacitor includes a standard capacitor and a metal shielding case in which the standard capacitor is installed.
Two terminations of the standard capacitors are respectively connected to input H-terminal and L-terminals of changeover switch through hermetical feedthrough, and the outer shell G is soldered to the metal case as common ground.
The changeover switch is installed in metal shielding case. Inside the case a metal shielding wall is installed to divide the case and switch into two compartments for H-terminal and L-terminal compartments respectively. In H-terminal compartment, there are multiple ceramics discs, and each disc has multiple contact positions, one disc is connected to output H-terminal and other 4 discs are connected to 4 input H-terminals respectively. In L-terminal compartment, there are multiple ceramics discs, and each disc has multiple contact positions, one ceramics disc is connected to output L-terminal and other 4 discs are connected to 4 input L-terminals respectively.
The decade capacitor standard has 6 changeover switches to select capacitance values. The changeover switch can rotate 360 degree.
FIG. 6
The changeover switch has open and short positions on one disc in both H-terminal compartment and L-terminal compartment for the compensation of short and open circuits. As shown in , one disc has OH and SH terminals in H-terminal compartment, and one disc has OL and SL terminals in L-terminal compartment. SH and SL are connected together for short circuit compensation, and OH and OL are unconnected for open circuit compensation.
1. The decade encoding of capacitances employs 4 capacitors and a changeover switch to realize decade capacitance output. The combination selected from 1 to 4 capacitors can output decade capacitance values. There are only 8 wires for connecting 4 capacitors to changeover switch. It significantly simplified the switch structure, and reduces the number of capacitors and connection wires comparing to existing decade capacitor standard box that have 10 capacitors. Therefore the changeover switch and decade encoding of 4 capacitors are optimized for compact size and better shielding to improve the accuracy and repeatability.
2. The changeover switch includes short and open positions within the switch structure, which enable to compensate the influence of internal connection wires and external test cables of RLC bridge meter simultaneously. This can remove all parasitic in the test system. It is easy for operation and can improve measurement accuracy and repeatability.
3. The decade encoding of all the combinations of 4 capacitors can reduce the number of required capacitors, the volume and weight of capacitor standard box, which is convenient for transport and field application.
Note:
FIG. 2
2
3
4
5
In : —changeover switch body, —Output ports, —Input terminals of changeover switch, —Output terminals of changeover switch
FIG. 3
1
2
3
In : —capacitor; —Changeover switch, —Outputs of decade capacitor standard box
FIG. 4
6
In : —Shielding metal box of capacitors, G—Ground
FIG. 6
21
22
23
4
5
In : —Ceramics disc switch; —Shielding metal box; —Shielding metal wall; —Input terminals of changeover switch; —Output terminals of changeover switch.
The technical proposal of the invention is described in details with drawings and embodiments in this section. The application of the invention is not limited to the disclosed here. Any new application derived from this invention belongs to the invention.
1
2
3
1
1
1
The invention is a high precision decade capacitor standard box. It comprises 6 sets of physical capacitors , six changeover switches , and output ports . Each set of physical capacitors includes four physical capacitors , at least one of the physical capacitor is a single-unit capacitance.
2
2
4
5
3
The changeover switch implements the decade encoding of all the combinations of 4 capacitors. The changeover switch has input terminals (H and L) that connect to two electrodes of physical capacitors respectively, and output terminals that connect to the output ports of capacitor standard box to provide the selected capacitance. The capacitance values can be selected in any integer from 0 to 10 times of single-unit capacitance.
As one example of all possible 4-capacitor sets, 4 capacitors can have a set of values as (1, 2, 2, 5) times of single-unit capacitance. This set of 4 capacitors can be encoded to output decade capacitances by changeover switch as shown in Table 2. 4 capacitors need only 8 wires for connecting them to a changeover switch. The changeover switch with simple structure can reduce the parasitic. There are other of possible 4-capacitor sets such as (1, 2, 3, 4), or (1, 1, 3, 6), etc. of single-unit capacitance that can be employed for decade capacitance output, which are protected by the invention.
1
The six sets of decade capacitors have unit capacitances of 100 nF, 10 nF, 1 nF, 100 pF, 10 pF, and 1 pF respectively. It is easy understood that their unit capacitances can be changed according to the requirement. For example, the six sets of decade capacitors can be chosen as 100 μF, 10 μF, 1 μF, 100 nF, 10 nF, and 1 nF.
3
H
H
L
L
The output port of decade capacitor standard employs 4-port BNC and 3-port BNC outputs, which includes H-port and L-port. The changeover switch has H-terminal and L-terminal outputs. The H-terminals and L-terminals of six changeover switches are connected respectively in parallel to the H-port and L-port outputs of capacitor standard box. Furthermore H-port output is connected to V-port and I-port outputs, L-port output is connected to V-port and I-port outputs.
6
6
The decade capacitor standard includes metal shielding case , and the standard capacitors are installed inside the metal shielding case .
6
6
Two electrodes of each capacitor are connected respectively to the H-terminal and L-terminal inputs of the changeover switch via feedthroughs, and the metal case G of each capacitor is soldered to metal shielding case by using a metal strip.
2
22
21
23
22
The changeover switch comprises a metal shielding case , multiple-ceramics-disc switch . A metal wall separates the metal shielding case into two compartments, and divides multiple-ceramics-disc switch into two equal parts, which are used for H-terminal and L-terminal inputs respectively.
21
5
4
The multiple-ceramics-disc switch has multiple discs in the two compartments. There is one disc to connect to the output terminal in each compartment. The other 4 discs connect to the input terminals respectively in each compartment. Although preferred embodiment is illustrated herein as a specific example that each disc switch is used for one capacitor input connection, other embodiments and examples, such as one disc switch being used for multiple capacitor connections may perform similar functions and/or achieve like results.
The high precision decade capacitor standard box has 6 dials that control 6 changeover switches to select the output capacitance.
The changeover switches can rotate 360° to select output capacitance values and On/Off in a step of 30°. The rotation step can be adjusted to other than 30°.
The changeover switch has a compensation function for open and short circuits. The 6 dials have two positions corresponding to the open and short status.
21
21
One ceramics disc in H-terminal compartment of the changeover switch has OH and SH contact points. Another ceramics disc in L-terminal compartment of the changeover switch has OL and SL contact points. SH and SL are connected together for short circuit compensation, OH and OL are unconnected for open circuit compensation. These two compensation processes can remove the parasitic from the input ports of RLC Bridge, testing cables, and internal wires and connectors of the capacitor standard. Comparing to existing compensation method, the novel design is convenient for operation, and improves the accuracy and repeatability in high precision measurement and calibration.
TABLE 1
Output × 100 pF
0
1
2
3
4
5
6
7
8
9
10
Input
100 pF
0
1
1
1
1
1
1
1
1
1
1
100 pF
0
0
1
1
1
1
1
1
1
1
1
100 pF
0
0
0
1
1
1
1
1
1
1
1
100 pF
0
0
0
0
1
1
1
1
1
1
1
100 pF
0
0
0
0
0
1
1
1
1
1
1
100 pF
0
0
0
0
0
0
1
1
1
1
1
100 pF
0
0
0
0
0
0
0
1
1
1
1
100 pF
0
0
0
0
0
0
0
0
1
1
1
100 pF
0
0
0
0
0
0
0
0
0
1
1
100 pF
0
0
0
0
0
0
0
0
0
0
1
Note:
0-not connected,
1-connected
TABLE 2
Output × 100 pF
0
1
2
3
4
5
6
7
8
9
10
Input
100 pF
0
1
0
1
0
0
1
0
1
0
1
200 pF
0
0
1
1
1
0
0
1
1
1
1
200 pF
0
0
0
0
1
0
0
0
0
1
1
500 pF
0
0
0
0
0
1
1
1
1
1
1
Note:
0-not connected,
1-connected
DESCRIPTION OF DRAWINGS
FIG. 1
1. is the front panel of the high precision decade capacitor standard box.
FIG. 2
2. is the circuit diagram of the high precision decade capacitor standard box.
FIG. 3
3. is the structure block diagram of the high precision decade capacitor standard box
FIG. 4
4. is the capacitor outline of the high precision decade capacitor standard box
FIG. 5
5. is the circuit diagram of changeover switch of the high precision decade capacitor standard box
FIG. 6
6. is the structure of changeover switch of the high precision decade capacitor standard box | |
Natural Ways to Lower Blood Pressure
The risk factors for developing high blood pressure are age, obesity, less active lifestyle, poor diet, and excess consumption of alcohol and tobacco products. Some of the effective ways to lower blood pressure are maintaining weight, regular exercise, stress management, healthy diet, low sodium intake, limiting alcohol consumption, and avoiding tobacco.
Ningthoujam Sandhyarani
The blood circulatory system is responsible for purifying oxygen depleted blood and supplying oxygen rich blood to the body organs. In the process, blood is continuously pumped in and out of the heart, thus exerting pressure in the arterial and venous walls. Blood pressure can be defined as the force exerted by the circulating blood against the walls of the blood vessels. It is considered as one of the most vital signs for determining the overall health and well-being of an individual.
Blood Pressure: An Overview
The amount of pumped blood and resistance to arterial blood flow are factors that determine the blood pressure. While assessing the blood pressure, there are two forces that are taken into consideration, namely, systolic blood pressure and diastolic blood pressure. The former corresponds to the pressure exerted when the heart contracts or pumps blood to the body cells and tissues, while diastolic blood pressure is the force exerted between two beats or when the heart is at rest.
Blood pressure is measured by using a device called sphygmomanometer, with its value expressed in millimeters of mercury (mmHg). It is represented as a fraction, with the systolic blood pressure at the numerator and diastolic at the denominator. A blood pressure reading lower than 120/80 mmHg is considered normal, while a lower or higher figure generally implies hypotension or hypertension respectively.
Lowering Blood Pressure Naturally
In the present scenario, high blood pressure, also known as hypertension is a major health problem worldwide. Though the exact causes of high blood pressure are not known, medical research reveals that lifestyle or the way of living contributes in developing hypertension to a great extent. Some of the risk factors for hypertension include age, poor diet plan, excessive alcohol intake, lack of physical activities, and obesity.
The following are certain natural ways to lower blood pressure that can be undertaken in order to avoid the complications associated with high blood pressure.
Maintain Weight
High blood pressure is related to excess weight or obesity. A majority of obese people suffer from hypertension. Losing weight and/or maintaining weight to normal recommended levels will effectively lower blood pressure.
Perform Physical Activity
A sedentary or less active lifestyle is a major risk factor for causing high blood pressure. Regular exercise, at least 30 minutes per day is recommended to bring down the blood pressure to normal, safer levels.
Manage Stress
Scientific studies claim that stress also contributes in developing high blood pressure. Hence, managing stress by practicing healthy coping methods, like, yoga and meditation help in combating hypertension.
Consume Healthy Diet
Changing unhealthy dietary and eating habits is effective in lowering blood pressure. Consume healthy foods which include fresh fruits, vegetables, whole grains, and foods containing low-fat and low cholesterol.
Reduce Sodium Intake
Excess sodium content in the diet can elevate blood pressure levels. Hence, cutting down on sodium intake as much as possible helps in reducing the blood pressure up to 2 - 8 mmHg. The more the amount of sodium reduced, the lower will the blood pressure be.
Limit Alcohol Consumption
Alcohol, if taken in excess amount, increases the blood pressure and reduces effectiveness of hypertension medications. Considering this, it is always advisable to limit alcohol consumption in order to lower the blood pressure levels.
Avoid Tobacco
Another factor contributing to high blood pressure is the nicotine present in tobacco products. Tobacco narrows the blood vessels, resulting in hypertension. Hence, cessation of smoking and other tobacco products will greatly help in lowering blood pressure.
In addition to these methods, other natural ways to lower blood pressure is getting adequate sleep, consuming fiber-rich foods, and reducing caffeine intake. It is also advisable to regularly monitor the blood pressure as doing this will help in proper planning of the diet and exercise in order to control the blood pressure to normal, recommended levels.
Disclaimer: The information provided in this article is solely for educating the reader. It is not intended to be a substitute for the advice of a medical expert. | https://holisticzine.com/natural-ways-to-lower-blood-pressure |
In the future, this page will serve as a forum for discussion, including an interactive blog feature.
For the present, I will use this space to present a few arguments showing why I think a “young earth” viewpoint is not the only Biblical explanation for the origin of the universe. These arguments were too detailed to include in Journey Through a Jewel.
In my opinion, proponents of a young earth, although sincere and zealous, may have done a great deal to harm the credibility of Christianity in the scientific world. By clinging to a narrow interpretation of Genesis Chapter 1 and, subsequently, appearing to force the geologic record into a mold supporting that interpretation, they may have created serious problems to combining good religion and good science. Unfortunately, young earth arguments are often the only ones presented as an alternative to a completely naturalistic theory.
On the other hand, scientists, such as John Clayton, Hugh Ross, William Dembski, Michael Behe, Jonathan Wells, and many others have presented compelling scientific evidence supporting “Intelligent Design.” However, many scientists mistakenly view the Intelligent Design argument as nothing more than Young Earth Creationism in new clothing. While it is true that evidence of fine-tuning in the universe and numerous examples of "irreducible complexity" found in many life forms has led some to accept the Intelligent Design theory, it does not necessarily lead to acceptance of the Bible or to religion in general. However, for those who do recognize the amazing compatibility between modern science and an old earth interpretation of Genesis Chapter 1, this evidence is faith strengthening and presents a viable alternative to Young Earth Creationism. For instance, on pages 233 to 239 of The Source by John Clayton and Nils Jansma (Howard Publishing, 2001) many fallacious young earth arguments for the formation of the geologic record and the Grand Canyon are discussed. I have written a Workbook for the Source which you can review for yourself. | http://sljansma.com/FrameElements/LowerFrames/Discussion.htm |
FanDuel, Inc. v. Interactive Games, LLC, No. 19-1393 (Fed. Cir. 2020)Annotate this Case
Interactive’s patent describes a gaming system wherein a gaming service provider—such as a casino—wirelessly communicates with users’ mobile devices, allowing them to gamble remotely. The system stores rules to determine the “game configuration” based on the location of a user’s “mobile gaming device” and associates different gaming configurations with different locations, using a “lookup table.”. FanDuel petitioned for inter partes review (IPR) of the patent on several grounds of obviousness. The Patent Trial and Appeal Board found unpatentable all challenged claims except claim 6, finding that FanDuel failed to prove that claim 6 was obvious in view of asserted prior art.
The Federal Circuit affirmed, rejecting a claim that the Board violated the Administrative Procedure Act by basing its finding on obviousness issues that Interactive did not raise in its responses. The Board’s purported new theory was merely an assessment of the arguments and evidence FanDuel put forth in its petition. The APA does not require the Board to alert a petitioner that it may find the asserted theory of obviousness lacking in evidence before it actually does so, nor is a petitioner entitled to a pre-decision opportunity to disagree with the Board’s assessment. The obviousness findings are supported by substantial evidence. | https://law.justia.com/cases/federal/appellate-courts/cafc/19-1393/19-1393-2020-07-29.html |
The role is challenging, varied and evolving. As a member of the senior business management team you will work in collaboration with the MD and senior management team to ensure business decision making. You will be directly accountable for functional performance and fully involved in all senior management/board discussions & decision making associated with the business.
The successful incumbent will shape and develops supply chain strategy and contributes to the development of business strategies.
Review and delivers supply chain improvements to meet income, margin and cost objectives and targets.
Provide leadership, direction, guidance, deployment of capability, governance and control of the supply chain function such that products and services are fit for purpose, safe and meet business, legal, regulatory and customer requirements.
Provide leadership of the functional management team and ensure the function has the appropriate resource and capability necessary to meet the needs of the business.
Ensure budgets and forecasts are in place and monitor expense (and income) such that financial performance is maximised and obligations to group are achieved.
Build strong relationships within the supplier base and the market in general, negotiate and agree alliances and partnerships to the longer-term benefit of the business.
Ensure processes are in place to carry out external benchmarking and establish external best practice and where appropriate implement revised operating methods and approaches to deliver continuous improvement to Supply Chain performance Compile, present and disseminate business information to Directors and the Board, based on effective supply chain services to inform performance issues associated with the supply chain.
The role carries immediate accountability for the Project Procurement & Supply Chain (P&SC) elements, spanning some £800m+ of planned expenditure and subsequent acquisition over the next 6 years. Key objectives include
* Ensuring commercial risk is appropriately shared with suppliers for the entire project and support services, with robust contracts, pricing and defined deliverables.
* Management of all supplier deliverables both paper and physical
* Addressing Strategy changes in Make-Buy, contract flow downs and performance management, resource deployment as the program progresses
* Supplier Relationship Management to ensure sustainable performance and develop the UK Prosperity Agenda
* Correct and disciplined use of a developing suite of management systems for purchase orders and material control
* Continuous Value Improvement and Supply Chain Risk Management
* Leading a team of high calibre professionals, some of whom are relatively inexperienced in construction programs.
A solid background in supply chain, you will have demonstrated progression throughout your career to date and have had sector experience in Engineering, Defence, Manufacturing,Oil & Gas, Construction. Candidates currently out of this sector at this stage, will not pass the selection process
Reference: 214703739
Set up alerts to get notified of new vacancies. | https://jobs.careeraddict.com/post/37195826 |
The next day, on January 19, HHS released a proposed rule that would provide protections for health care workers who refuse to participate in services that run counter to their religious beliefs or moral convictions. Finding support in the First Amendment and federal anti-discrimination statutes, the proposed rulemaking is intended to protect doctors and nurses with moral or religious objections to participating in certain procedures such as abortions, assisted suicide, sterilization, vaccination, and other matters of conscience. Construed broadly, the rule may also cover conscience objections to treating transgender patients or issues of gender identity.
The proposed rule applies to entities that receive funds through programs funded or administered in whole or in part through HHS, such as hospitals, nursing homes, clinics, physician offices, universities, and more. Public comments are now being accepted on the proposed rule through March 27, 2018.
The rule likely will lead to increased scrutiny from OCR and the newly formed Conscience and Religious Freedom Division. The Washington Post has quoted OCR Director Roger Severino about the more than three-fold increase in the number of worker complaints related to moral and religious beliefs since President Trump’s election. Severino stated that a career senior executive will be appointed to investigate such issues. Under the rule, OCR would have the authority to initiate compliance reviews, conduct investigations, coordinate compliance, and use enforcement tools comparable to those available under other civil rights laws.
Once finalized, the rule will have an immediate impact on how health care providers assign employees, discipline for failures or refusals to perform certain services, and accommodate religious and moral beliefs. The requirements may give rise to issues that will need to be addressed in collective bargaining agreements. While we expect there to be legal challenges to the rule, employers should be prepared to both address employee concerns about providing services to which they have conscience objections, and to create alternative plans for treatment when such concerns are raised. | https://www.healthemploymentandlabor.com/2018/01/26/hhs-moves-to-strengthen-conscience-protections-for-health-care-workers/ |
The Geatest Common Divisor of two positive integers is the largest integer that divides the two integers with no remainder. The GCD is useful for reducing fractions to their lowest terms.
The file gcd.cc contains the outline for a program that reads in a
pair of integers from the user, passes the entered integers to the the
undefined method
GCD, and prints the result. The
GCD method should use a
brute force algorithm to find the greatest common divisor of the two specified
integers. The method can assume that the two parameters are positive integers,
but should have a method comment that specifies this assumption. Be sure to
test your code on multiple examples and on the cs server.
Tar your code in a file that contains your name and submit it on the course Inquire site. | http://cs.roanoke.edu/Fall2011/CPSC250A/assignments/assn0/assn0.html |
The comprehension and production of language, common and effortless in everyday life, require the coordination of multiple linguistic and cognitive processes with millisecond accuracy. Some of these language components include speech perception, echoic verbal memory, lexical-storage, lexical retrieval, associating words with related words and concepts, applying grammatical rules, finding phonemic representations for words, and planning and executing articulator movements. New technologies support the investigation of the precise neural structures that support these components and the time course of their interactions with ever-greater precision. The goal of this project is to investigate the brain areas involved in three components of language for which our previous work has isolated brain regions that appear to contribute to these functions: articulation, lexical semantics, and executive control of language processing. In the proposed study, we will investigate the specific functional roles of these brain areas, the time course with which they are recruited into the language system, the ways in which these areas interact with each other, and how they interact with other areas that support the cognitive skills necessary for language (e.g., memory and executive functioning). Our proposal uses three technologies available to us in our group, lesion analysis, functional NMI, and event related potentials. Lesion analysis will be used to correlate components of language behavior of aphasic patients who exhibit deficits with the specific brain regions affected. Because lesion analysis isolates the area that disrupts a particular function, but does not specify its role in real-time processing, functional MRI experiments will parallel the tasks used with aphasic patients but will investigate the functional roles of these areas in language use by normal subjects. ERP studies will be used to examine the time course of lexical selection and executive control processes to determine the points at which they enter into language use and when they interact with other cognitive functions. Together, this rare combination of techniques will teach us more about the brain areas involved in speech and language, their functional contributions, and the time course of their involvement.
| |
Our experienced estimators complete their take offs and scope clarifications thouroughly on the front end considering transitions that minimize punch list items and warranty call backs, product application combined with quality setting materials to deliver a job that will last and not fail over time. Experienced estimation minimizes waste and drives points to the bottom line of our builders.
Prewalk Process
Our production team walks each job after subfloor and framing is finalized to determine jobsite rediness two-weeks prior to flooring to ensure enough time to discuss corrections needed prior to flooring to ensure a quality install per manufacture specifications per product selection.
Lessons Learned
Our production team meets weekly to discuss “lesson learned”. A review of each production case from the week prior is reviewed and discussed as a team. The result is feedback internally with a colloborative improvement strategy to avoid repeated issues and costs associated. This drives effeciently resulting in improved quality of work combined with improved pricing. | https://preferredfloorandtile.com/home-builders/ |
The Northern Institute team, of nationally and internationally recognised researchers, are leaders in social and public policy research.
Teams work to develop a series of activities in partnership with stakeholders and funding bodies on areas of key interest for the region, engage nationally and internationally recognised to lead that work and dissemination activities.
The teams are supported to target partnership activities with internal and external agencies that promote social and public policy research and public debate based on sound research.
Each undertake a cohesive set of work that utilizes national and international partnerships and funding opportunities to build capacity and opportunities of high quality public policy research.
Principal scientists in key research areas grow the capacity, improve partnerships and networks and invest in a legacy through their work.
Mid and early career researchers are linked to principal researchers to develop their research skills and capacity to participate in high level research and publication.
It is the responsibility of these research teams to promote the work of TNI’s researchers, connect to regional and national priorities and explore new methodologies of research and dissemination.
Research outcomes build on the strengths related to living in remote, regional and urban regions in the northern region.
Research support
Increasing support for research demonstrates the impact of that collaboration, as agencies have been able to identify the beneficial outcomes of research collaboration and CDU and its partners have improved their ability to negotiate research with a strong local impact.
The Northern Territory Government and Charles Darwin University have developed a Partnership Agreement that provides a framework to manage and improve collaboration for research outcomes.
This partnership has enabled investment in research capacity building approaches to undertaking research, supporting internal research partnerships across CDU to flourish and identifying local talented researchers of the future.
CDU has developed MOUs with regional universities and industry partners. The three Cooperative Research Centres with which CDU is involved provide essential opportunities to collaborate with a range of university and industry partners, this is evident in the demography, health and education policy spheres.
Collaboration for the future
The collaboration with Aboriginal communities over decades is evident in the research with people in the Tiwi Islands and East Arnhem land. Similarly the long term partnerships with people in East Timor over time and change is evident in the body of work undertaken in the region with local people who are developing as early career researchers while also being senior knowledge holders in their communities.
The Northern Institute's key alliances and partnerships include:
- Innovative research universities
CDU is one of seven members in the Innovative Research Universities network, through which the members direct their expert knowledge, capabilities and resources to enhance the outcomes of higher education. CDU sits alongside Flinders, Griffith, James Cook, La Trobe, Murdoch and Newcastle universities in the network.
- Key university partners
Australian National University, Flinders University and James Cook University enjoy an important organisational partnership with CDU, which enables The Northern Institute to enter into specific research relationships, joint projects and the joint supervision of students
- Northern Territory Government
The Territory relies on a robust and responsive university to help drive its economic and social development. The Northern Territory Government and CDU Partnership is a formal agreement that is unique for an Australian university and supports education, research, policy and development and program delivery.
- Non-government sector
Across Northern Australia, the non-government sector plays a critical economic and social development role in the governance and service delivery to rural and remote communities. The Northern Institute works collaboratively to deliver research, strategic advice and training to this important sector.
- Industry sector
As the communities of Northern Australia prepare for rapid growth in minerals, oil and gas exploration, construction and demographic change. TNI is well placed to deliver high quality advice, research and development, as well as short courses training to the sector.
- Marine science
The Australian National University, the Australian Institute of Marine Science, the NT Government and CDU are members of innovative partnership that delivers cooperative research, academic and training activities, and expanded research infrastructure, to ensure sustainable and community development of tropical marine resources. | https://www.cdu.edu.au/northern-institute/our-partners |
Doha: The Ministry of Municipality and the Ministry of Environment and Climate Change launched the English version of 'Eco Tourism in Qatar' at the 31st Doha International Book Fair yesterday.
The book written by Ahmed Hussein Al Mutawa, the environmental consultant at the Ministry of Environment and Climate Change, was published in Arabic in 2019.
The event was presented by Engineer Farhoud Hadi Al-Hajri, Director of Public Relations at the Ministry of Environment and Climate Change.
Addressing the event, Al Hajri highlighted the importance of the book and its role in raising environmental awareness and informing society of the efforts made by Qatar in the field of preserving the environment and ensuring its sustainability in line with Qatar National Vision 2030.
He said that the participation of the two ministries in this year's book fair is part of their efforts to spread awareness and community communication through the presentation of various books, literature and publications carried out by the Ministry of Municipality and the Ministry of Environment and Climate Change, in addition to introducing their achievements and the outcome of their work through the presentation of documentary films and publications.
For his part, Ahmed Hussein Al Mutawa spoke about his book 'Eco Tourism in Qatar', highlighting the great role that this type of tourism enjoys at all levels in the country.
The writer explained that the tourism industry is witnessing a tangible and rapid development, as the State has developed a comprehensive tourism plan to keep pace with the economic renaissance, and to emphasise building Qatar as a global destination.
The discussion session, which was attended by a number of environmental officials and activists, witnessed a rich and fruitful discussion, which touched upon a number of issues and problems related to the environment and tourism and their relationship to economic development and the urban development witnessed by Qatar, and discussed ways to achieve balance and reach sustainable development while preserving natural resources for future generations.
MENAFN22012022000063011010ID1103577104
Legal Disclaimer: MENAFN provides the information “as is” without warranty of any kind. We do not accept any responsibility or liability for the accuracy, content, images, videos, licenses, completeness, legality, or reliability of the information contained in this article. If you have any complaints or copyright issues related to this article, kindly contact the provider above. | https://menafn.com/1103577104/English-version-of-Eco-Tourism-in-Qatar-launched-at-DIBF&source=30 |
Working at Malta means being a part of a community that values growth, mutual trust and respect.
We want to continuously learn and improve, so we maintain a feedback-oriented mindset. And most importantly, we care – about creating sustainable solutions, about how our actions affect the world, and about each individual employee’s experience. We offer a safe, inclusive and productive environment for all team members, and we’re always open to feedback.
Malta's Core Values
These values reflect what’s important to the entire company. They guide our efforts, define our culture, shape the way we approach our solutions, and change alongside us as we aim for more and more ambitious goals.
Exceed expectations
Never settle for "good enough" when you know you can and should do better.
Take ownership and question the status quo in a constructive manner
Trust your gut and your knowledge, even if you need to question your team leader or CEO.
Be brave, curious and experiment – learn from all successes and failures
Take risks and make mistakes - that’s how we learn, and how we innovate.
Act in a way that makes all of us proud
We are all in this together and everything you say and do, whether internally or publicly, reflects on us.
Build an inclusive, transparent and socially responsible culture
Culture doesn’t happen by chance – we need to be proactive and work on it every day.
Recognize excellence and engagement
We know the value of well-earned respect and constructive feedback.
Current Openings
There are currently no openings. Please check back soon! | https://maltainc.com/company/join-our-team/ |
Basic & Applied Science
The School with all its facilities promises an overall development of student community in terms of both intellectual and personality development.
Overview:
The School of Basic and Applied Sciences (SBAS) of Dayananda Sagar University (DSU) stands up to the task of producing world class graduates with capability and confidence to handle real-life challenges. The School with state-of-the-art facility imparts quality education and conducts cutting edge interdisciplinary research in science through its various disciplines. Basic Sciences help a student in comprehending the fundamentals of science and technology; it creates interest and helps to nurture the young mind in preparation for the challenges ahead. Application oriented subjects on the other hand, facilitates the development of intellectual prowess, impeccable skill sets and create the ability to apply oneself efficiently to tackle day to day intricate problems.
The curriculum at SBAS focuses both on developing the intellectual appetite of the students as well as helping them to become skilled at novel problem-solving approaches. The School allows the flexible Credit-based Choice system, which maximizes the potential of the student and facilitates teacher-student interactions. This system allows the mobility of the products of the school both within and across the geographical jurisdiction.
The School of Basic and Applied Sciences has pooled an excellent group of highly qualified, experienced and research-driven teachers that provide the platform needed for the novice student to blossom into an educated, cultured and highly trained professional suitable for industry and scientific organization. The school has been provided with state-of-the-art laboratories and well-equipped libraries that cater to the needs of the students. Emphasis is laid on continuous evaluation of students.
The School of Basic and Applied Sciences regularly conducts workshop for students to gain hands-on experience on advanced techniques, quiz, seminars, organizes guest lecturers, international and national conferences, apart from inter-collegiate and inter-class competitions. It also encourages the students to take part in national and international conferences to present their research papers.
Program Educational Objectives:
- Unity in diversity: to foster students from a diverse culture and bring them together for problem solving.
- Problem solving: to engage in research activities leading to creation of tools and technologies to address the problems of the society.
- Enhance the University's scientific, social and cultural impact through engaging in research, conferences, workshops and publications in reputed journals.
- To create an environment that nurtures individual as well as group work, but more importantly that allows scope for creative sparks. | https://www.dsu.edu.in/basic-applied-sciences/life-sciences |
We cannot alter the genes we are born with — but that definitely doesn’t mean we are irrevocably set on a path we can’t change when it comes to whether or not we go on to develop cancer.
As a consultant oncologist, I have studied the causes and consequences of cancer for most of my professional life — and I am now convinced that the way we choose to live our lives, day in, day out, can make a much bigger difference to our health and wellbeing than the genes we inherited.
The more research I undertake and the more patients I see whose health has been immeasurably improved by changing their diet and lifestyle, the stronger my conviction becomes.
We cannot alter the genes we are born with — but that definitely doesn’t mean we are irrevocably set on a path we can’t change when it comes to whether or not we go on to develop cancer
That’s why I wrote my new book, How To Live, in the hope of sharing the experiences and advice based on my lifetime’s work.
And it’s not just one individual factor that counts, as I explained in Saturday’s paper, but the sum of all the little choices that we make every day over the years. The food we eat (and how we prepare it) plays a very significant part in our health.
More and more studies are now linking poor gut bacteria with the risk of several cancers — both in and outside the gut — which is why, as we continue our exclusive series this week, today I’ve chosen to focus on our digestive systems.
What’s more, poor gut health doesn’t just cause problems in your digestive organs — it’s the cornerstone for many other serious diseases in the rest of the body, including diabetes, obesity, Parkinson’s disease and arthritis.
(And that’s not to mention bloating and indigestion, problems with bowel movements, low mood, sleep disorder and even depression, which can also be rooted in poor gut health)
Based on the latest research, I’ll explain just why your gut health matters and show you practical ways to build a thriving community of good gut bacteria that can help you reduce your risk of cancer and other serious diseases.
Don’t forget to brush your teeth
The process of digestion starts in the mouth, home to hundreds of species of bacteria.
The bacteria in and on our bodies — and particularly in our digestive systems (or microbiome) — can roughly be divided into ‘bad’ (those that cause infections such as food poisoning or cholera and typhoid) and ‘good’ (bacteria which prevent the growth and spread of disease, improve overall immunity and help to reduce chronic inflammation, which occurs when our bodies mistakenly react as if they are permanently under threat from disease).
Good bacteria are generally referred to as probiotic bacteria and include the lactobacillus and Bacteroidetes groups, linked with numerous health benefits. (I’ll look later at ways you can boost your good bacteria.)
However, poor dental hygiene — caused by not brushing your teeth or not flossing correctly — can upset the balance between the two, leading to a build-up of harmful bacteria, causing gum disease and tooth decay.
Chronic inflammation of the gums (or gingivitis) is linked to a higher risk of chronic diseases elsewhere in the body, particularly dementia, diabetes, heart disease and emphysema.
In terms of cancer, we saw on Saturday how two studies analysing more than 100 samples of healthy and cancerous bowel tissue found that DNA from bacteria commonly found in dental cavities was also present in bowel cancer tissue — but not in normal, healthy cells.
This led researchers to conclude that bacterial DNA from the mouth had travelled through the body, interacting with and being absorbed into gut cells, causing them to become cancerous and leading to bowel cancer.
Boost your ‘good’ bacteria
These days we read a lot about why we need strong, healthy colonies of good bacteria.
And rightly so — I cannot stress enough how important it is to cherish and feed your good bacteria because they have a vital role to play in keeping your immune system in mint condition.
This is key to helping our bodies detect and destroy early cancer cells more efficiently as well as any carcinogens in our food and environment, as I explained in Saturday’s paper.
It’s also well established that poor gut health contributes to numerous digestive problems including bloating, irritable bowel syndrome (IBS), diarrhoea, food allergies and intolerances.
Research now shows that a depleted colony of good bacteria leads to damage or thinning in your gut walls, weakening the barriers that normally prevent toxins (including carcinogens) from getting into the blood stream.
As a result, these toxins and bacteria can cause inflammation all over your body. This is commonly known as ‘leaky gut syndrome’. But it doesn’t end there.
An inflamed gut wall also makes the body less efficient at absorbing the vital nutrients it needs from food — so over the long term, this can cause deficiencies in vitamins A, D and zinc, all of which have important roles to play in our immune system’s defence against disease and cell damage.
Dining out? Then don’t overeat
Eating out and staying away from home can play havoc with your microbiome, as you are more likely to consume more alcohol and dine out on rich or unaccustomed foods.
To look after your gut when dining out or going away, try not to overeat and try to have alcohol-free days when you are on holiday.
Eating lots of fruit and vegetables and drinking plenty of water should keep your bowels regular, but if you are constipated you could try taking some extra flaxseed (also known as linseed).
In addition, many people, including me, swear by the protective benefits of a good-quality probiotic supplement, starting a couple of days before travelling.
This is why a wide range of serious diseases in other parts of the body — including dementia diabetes, arthritis and obesity — can either be caused or aggravated by poor gut health and chronic inflammation.
There is also emerging data to suggest it is a ‘trigger’ for type 1 diabetes — because the body gets confused by toxins in the blood stream and starts inadvertently attacking its own pancreas.
This chronic inflammation also helps to explain why the tumours associated with poor gut health are not just restricted to several types of bowel cancer.
In addition, scientists now know that healthy colonies of good gut bacteria use phytochemicals to produce a fatty acid called butyrate that helps protect the cells lining the colon from genetic damage.
Not only this, but butyrate also kills established colon cancer cells before they get a foothold — yet another reason why eating a diet high in phytochemical-rich vegetables, fruit, herbs and spices is a key way to look after your gut health and cut your cancer risk.
The cancer-fighting results of feeding your good gut bacteria with phytochemical-rich foods was recently illustrated by an exciting academic study that I was involved with at the Primrose Oncology Research Unit at Bedford Hospital.
Known as the Pomi-T study (after the extracts we tested: pomegranate, broccoli, turmeric and green tea), it is to date the world’s largest and probably most respected trial evaluating the impact of phytochemical-rich foods.
It was clear from our trial, which involved 203 men with diagnosed prostate cancer, that taking a gut-friendly probiotic supplement rich in phytochemicals not only reduced inflammation in the gut but also slowed the progression of the prostate cancer in those who took it, compared with those in the control group who did not.
This is one of the many reasons I am so excited at how many ways there are, based on the latest science, to take control of our own health — starting with looking after our guts. It really does help to look at your diet again and think about ways to eat for your gut.
I’ll look in more detail at which supplements are helpful to take in tomorrow’s pullout.
Going organic really can be better for you
Pesticides and other chemicals make it into our food chain without us realising it — and some can be very damaging.
They are used to control insects, weeds, fungi and bacteria and help to ensure we can produce enough food to meet demand.
But inevitably they also make it into the food chain — and some (organochlorines and organosulfates) can cause cells to mutate, while DDT, chlordane and lindane are tumour promoters.
The pesticide MXC was developed after the ban of DDT but tests have also shown that this stimulates the proliferation of breast cancer cells. Its use is falling, but others with potential risks are replacing it.
Pesticides and other chemicals make it into our food chain without us realising it — and some can be very damaging
Some insecticides also contain arsenic compounds still in common use which have been classified as carcinogens by the International Agency for Research on Cancer (IARC).
Meanwhile, chlorothalonil, a fungicide used on trees, vegetables and agricultural crops, has been classified as ‘likely’ to be a human carcinogen.
Non-Hodgkin’s lymphoma, a cancer of the lymphatic system, has previously been associated with exposure to glysophate, a herbicide commonly used as weedkiller.
Although the evidence of risk is small outside farm workers, it’s certainly worth trying to reduce your intake of pesticides over time by buying organic, growing your own fruit and veg and making sure you wash any shop-bought produce thoroughly before you eat it.
Organic food cannot be completely free of synthetic chemical residues, due to product and environmental pollution, but organic agriculture avoids synthetic fertilisers and pesticides, and instead uses holistic methods of weed and pest control such as long crop rotations, natural predator insects and insect traps.
Organic meat also has lower levels of other potentially harmful contaminants, as livestock is not given growth hormones or antibiotics, so it is worth buying organic whenever you can.
8 ways to improve digestive health
Protecting your ‘good’ bacteria should be top of your to-do list when it comes to focusing on how you can cut your cancer risk by improving your gut health.
You can either do this by ‘topping them up’ with other living microbes called probiotics or by ‘feeding’ them with compounds that strengthen them.
Here are my eight top ways to protect your core asset — and reduce your risk of a number of cancers.
Say yes to chocolate
A square or two of dark chocolate may seem an indulgence but could also help towards a gut-friendly diet rich in phytochemicals.
Chocolate contains many polyphenols, as well as theobromine — a vasodilator which increases heart rate, lowers blood pressure and boosts brain levels of the mood-enhancing chemical seratonin.
One study found those who regularly eat dark chocolate tended to have lower blood pressure.
What about milk and dairy?
Many people wonder if dairy is good for them. This is partly because cow’s milk is a common cause of allergies and digestive problems, though these can usually be diagnosed with tests or elimination.
But there are some reports that milk and dairy products are carcinogenic.
I think it’s likely that the risks of moderate milk consumption have been overemphasised.
It is often quoted that Asian cultures have a low cancer rate as they don’t drink much milk, but other factors could account for this, including being more active and eating more fruit and vegetables.
There are no robust randomised trials linking milk with an increased cancer risk. In fact, higher milk intake has actually been found to reduce bowel cancer risk.
Analysis of 12 large studies showed no significant increase in ovarian cancer risk in those who consumed a moderate amount (one or two glasses). However, more than two glasses of milk a day did increase the risk in relapsing after treatment for breast and ovarian cancer.
Similarly, a large study recently linked high calcium intake (from milk and supplements) with a raised risk of prostate cancer, but concluded one to two glasses was not associated with increased risk.
As we’ve seen, kefir and live yoghurt are among the healthiest ways to consume dairy products (so long as you are not lactose intolerant) because they contain probiotics as well as being rich in protein, vitamins, omega fats and energy.
A cup of char can work like a charm
Do you feel as though you can’t face the day without a cup of coffee in your hand? If so then you can rest easy — at least as far as cancer is concerned.
Research shows that moderate consumption of coffee is actually good for you in terms of cancer and can reduce your risk of prostate, liver, womb cancer and some cancers of the throat and mouth.
This is despite the fact that the coffee beans are roasted — a process that produces moderate levels of acrylamides.
Baking, grilling or roasting foods at high temperatures produce these toxic compounds, linked to an increased risk of cancers.
Do you feel as though you can’t face the day without a cup of coffee in your hand? If so then you can rest easy — at least as far as cancer is concerned [File photo]
But the evidence of recent studies implies that this is substantially outweighed by the benefits from all the cancer-protective phytochemicals in coffee.
As for tea, the great British cuppa is not just a national treasure, it can also help to lower your risk of cancer.
Both green tea and the black stuff we’ve been drinking in the UK for hundreds of years contain high quantities of numerous polyphenols and it may surprise you to learn they both come from the same plant.
When the leaves of the Camellia sinensis plant are dried, they are fermented and oxidised to form the black tea we are familiar with. Green tea is unfermented and merely steamed.
Tea polyphenols are important because they have been shown to block an enzyme which tells cancer cells to proliferate faster and bypass a process called apoptosis, which is a pre-programmed self-destruct mechanism by which our body gets harmful cells to destroy themselves.
They also have excellent anti-inflammatory properties.
Despite these positive effects on biological pathways, not all human studies have linked a daily cuppa or two with a lower risk of cancer.
In fact, the large Japanese Ohsaki National Health Insurance Cohort Study reported that tea consumption was not associated with lower cancer levels, and a large Cochrane review concluded that there was insufficient evidence for a benefit or risk, although quality of life seems to be better in tea drinkers.
However, I am convinced of the anti-cancer benefits of tea drinking, partly from studying other data and partly because of a big research project that I was involved in.
Our research team at the Primrose Research Unit at Bedford Hospital undertook a 155,000-person analysis, which performed the world’s largest specific analysis of tea, following participants for 12 years. We were able to prove that drinking two to three cups of tea a day reduced the prostate cancer risk.
Interestingly, there wasn’t a benefit for those tea drinkers who added sugar to their cups, however.
Population studies have shown that people who drink tea regularly have lower risks of arthritis as well as Parkinson’s and Alzheimer’s, which is thought to be due to a reduction in the build-up of harmful amyloid in brain tissue.
The protection from heart disease and stroke by green tea (although not black tea) is thought to come from its ability to lower LDL (bad) cholesterol. Finally, regular consumption of tea has even been shown to improve bone health and reduce the risk of developing arthritis, as well as improving exercise performance.
So this is why, as a doctor, I wholeheartedly prescribe you to put the kettle on and treat yourself to a cup of your favourite brew!
Changing diet helped me cope with chemo
Julie Woodgate, 46, is a business analyst who lives near Windermere in the Lake District with her husband Martin Scovell, 60, a marketing executive.
Julie, who has a grown-up stepdaughter, changed her diet and lifestyle to help her to deal with chemotherapy and surgery after being diagnosed with stage 2 breast cancer eight years ago. Julie (below) says:
When I found out I had breast cancer I was determined to give myself the best chance I could of recovering from my treatment and living to enjoy as many more years as possible.
So I was keen to try any diet and lifestyle changes that might help me to cope with the side-effects of eight rounds of chemotherapy after being diagnosed with stage 2 breast cancer when I was only 38 in 2012.
On Professor Thomas’s recommendation, I ate much less red and processed meat — at first this was quite hard because I ate meat most days. Giving up my favourite, sausages, was the biggest sacrifice. I also used to love sweets — like a bag of pick and mix — but I stopped eating those too and cut right back on alcohol.
Instead, I drank lots of water, at least two and a half litres a day, and this helped to flush out the toxins in my chemotherapy drugs and reduce the severity of the nausea, hot flushes, constipation and terrible fatigue.
I also made a massive effort to eat lots of green, leafy vegetables. I took some phytochemical supplements and added turmeric to the recipes I prepared to maximise the amount of extra benefits I could get from all the polyphenols and other healthy plant chemicals.
Vegetables have now become such a part of my life that I’ve got my own veg patch where I grow purple-sprouting broccoli, peas, cauliflower, chard, beans and sprouts, which have the added benefit of being free from pesticides.
Another massive change was taking up exercise. Before my diagnosis, I didn’t do much but now I do three sessions a week with a personal trainer.
It turned out that, like the actress Angelina Jolie, I had inherited the BRCA gene mutation which made me likelier to develop breast cancer and at an earlier age. After studying my options, I chose to undergo a double mastectomy to minimise the chances of my localised cancer spreading to both breasts.
My cancer has been in remission since 2013 but in the hope of giving myself the best chance, I chose last year to also have my ovaries removed as a preventative measure. This has the effect of reducing the amount of oestrogen your body produces, which in turn lowers the risk of breast cancer returning.
Now I really live in the moment and I enjoy my healthy lifestyle. My stepdaughter’s expecting a baby soon — and I’m looking forward to being a granny.
Extracted by Judith Keeling from How To Live by Professor Robert Thomas, published by Short Books at £14.99. © 2020 Professor Robert Thomas.
Order a copy for £12.74 at mailshop.co.uk/books or call 020 3308 9193.
Free UK delivery on orders over £15. Promotional price valid until October 10, 2020. | https://wcrynews.com/world-news/why-keeping-healthy-is-all-about-guy-instinct/ |
Ten working groups organized under the Shaping Our Appalachian Region, or SOAR, initiative have presented their top findings. And despite the different working group topics, several common themes emerged among the recommendations:
• The need for cross-agency collaboration, especially in economic development
• Desire for training and investment in entrepreneurship
• Strong interest in a robust public relations effort to both market the region to the United States and beyond and to reinforce the citizens’ understanding and support of the uniquely Eastern Kentucky story
The recommendations – some familiar, some novel – are the synthesis of a summer spent talking with Eastern Kentucky residents in more than 100 meetings and listening sessions across the region. In all, more than 2,500 citizens participated in discussions about topics ranging from education to tourism to economic development.
The SOAR Futures Forum committee, chaired by former Gov. Paul Patton, met for the first time on Tuesday. The committee will focus on long-term strategies to improve the region. Members of this group will work with the executive committee and work groups to translate immediate and ongoing efforts into future strategies. The Futures Forum committee will also develop a SOAR 2025 Vision to be presented to the region next spring.
As a result of the SOAR Health Impact Series in August featuring Dr. Thomas Frieden, director of the Centers for Disease Control and Prevention, Kentucky Congressman Hal Rogers announced that Dr. Frieden has committed a full-time CDC senior staff member and additional support for the SOAR initiative for one year beginning in October.
The CDC representative, who will report to the SOAR executive director, will bring additional knowledge, expertise and experience in planning and managing public health activities. The staffer will be housed in the SOAR office.
Two organizations also announced financial support for SOAR. The Appalachian Regional Commission is pledging up to $750,000 over the next four years to underwrite half of the start-up expenses for SOAR administration, including the salary for the executive director. The Kentucky Association of Counties donated $5,000 toward SOAR operations.
Working Group Recommendations
Each group submitted a report with goals that could be reached in three separate time frames: within the next year, within one to three years, and within 10 years. Some highlights of the shortest-term recommendations follow.
Agriculture/Community/Regional Foods:
· Support local food system development through local education efforts, a national tourism campaign called Bon Appétit Appalachia, and allowing WIC and senior vouchers to be used at local farm stands.
· Compile information about agriculture, food, and natural resource asset mapping efforts.
· Create part-time position for agriculture liaison in SOAR administrative structure.
· Create low-interest loans for small and beginning farmers in Appalachian region.
· Consider tax incentives to lease reclaimed mine lands for agricultural purposes.
Broadband:
· Fiber infrastructure should be an open access system to support government services, education, health care and business development.
· Fiber infrastructure deployment should be a priority project for the SOAR region, followed by other regions in the state.
Business Recruitment:
· Identify regional growth zones that have emerged around growth communities within the region, based upon statistical metrics.
· Identify emerging economic clusters throughout the region for focused development.
· Identify existing companies that are growing, and focus programs of the eastern Kentucky Technical Assistance Providers Network on this group, via the development of regional business service teams.
· Develop and promote a web portal clearinghouse to better market resources currently available to potential entrepreneurs and existing small business owners.
· Begin to craft a multifaceted campaign to tell the story of innovative entrepreneurship within the region, especially to our youth.
Business Incubation:
· Begin formal studies with a consultant on several issues to include existing resources, properties inventory, workforce inventory and target industries analysis.
· Create new relationships among new and existing regional economic development agencies and engines, and provide funding for collaboration.
· Develop specific incentive programs for Eastern Kentucky.
· Improve critical infrastructure, including transportation, high-speed Internet and industrial park properties.
· Establish permanent economic development funding tied to coal severance funds.
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Education and Retraining:
· Establish an employment and training program focused on low-wage workers and unemployed individuals who qualify for Supplemental Nutrition Assistance Program (SNAP) benefits.
· Develop an entrepreneurial training program at the community colleges that supports the region’s rich artisan culture and small business owners.
· Work with partners in healthcare, energy, telecommunications, and digital technology to guide creation of short- and long-term training programs to establish direct paths to employment in those sectors.
· Connect education to training for the workplace. Increase access to education through additional career counselors, uniform career portfolios and partnerships with local businesses.
· Ensure educational and regional leadership through promoting leadership academies and access to professional development.
Health:
· Endorse and promote the passage of a state-wide smoke-free legislation.
· Start a “Healthy 5 for the 5th” campaign to promote wellness in the region.
· Explore Coordinated School Health programs for our entire region.
· Ramp up oral health efforts to encourage school-based oral health services.
Infrastructure:
· Complete a SOAR Transportation Planning Study for the region.
· Require participation in a regional planning process as a condition for funding in order to identify the most important and cost efficient solutions or projects.
· Promote utilization of MACED’s How$mart Program and other existing programs.
· Reform the financing, permitting, and policing of water and sewer systems within the SOAR region to improve service.
· Create a SOAR economic development organization.
Leadership Development and Youth Engagement:
· Create county coalitions focused on the empowerment of eastern Kentucky young workers, which will support training and professional development and create regional networking and social opportunities.
· Promote entrepreneurship, SOAR and specific change strategies including angel investment and coding effort; engagement with schools; and early entrepreneurial education.
· Create a SOAR student voucher program for area cultural events for low income children and a parent or guardian to attend one event or attraction each semester.
· Create a positive awareness campaign for youth using area magazines, newspapers, radio, and television opportunities to help share positive eastern Kentucky stories.
· Sponsor local college and career fairs through area high schools.
Regional Collaboration and Identity:
· Use Area Development Districts as supporting entities for development and administration of projects in ARC counties.
· Establish platform to insure that communication continues among work groups, local governments, private business and interested citizens.
· Encourage existing economic development professionals and organizations to maximize their strength by collaborating on marketing and recruitment of relevant industry.
· Utilize existing resources to create the framework for regional foundations that mirror regions as defined by area development districts.
Tourism, including Natural Resources, Arts & Heritage:
· Increase funding for aggressive advertising and media outreach, to include strategic marketing, public relations, and social media to effectively brand Kentucky and Appalachia and market the area nationally and internationally.
· Promote heritage and the arts of the region (visual, performing, and literary) through more artisan centers; restoration of “dying” traditions in heritage and the arts; offering business training for artists; and supporting more heritage tourism.
· Promote entrepreneurship through incubator support and mentoring programs.
· Promote SOAR region through smartphone apps, online promotion and social media.
· Chart existing festivals and cross-promote; provide festival training on how to maximize return on investment. | https://www.kyforward.com/new-ideas-new-partnerships-emerge-as-soar-groups-share-top-ideas-for-eastern-kentucky/ |
Self-monitoring has been proven to be effective in increasing performance, on task behavior, and homework completion in students with learning disabilities and other academic difficulties. Parent supervision and involvement in homework also has been shown to improve performance. However, the effectiveness of parent vs. self-monitoring of homework completion and test preparation has never been examined in adolescents with Attention-Deficit/Hyperactivity Disorder (ADHD). The current study compared the effectiveness of self- and parent-monitoring of homework and study skills completion in middle school students with ADHD. Students were trained in the SQ4R study strategy and homework completion skills. In one group students monitored their own behavior and in the other group parents completed the monitoring. Homework and classroom problems were evaluated using the Homework Problem Checklist (HPC) and the Classroom Performance Survey (CPS). Results indicated that both interventions improved HPC and CPS scores, as well as percent of completed homework.
Date
2005
Document Availability at the Time of Submission
Release the entire work immediately for access worldwide.
Recommended Citation
Meyer, Kara J., "Improving homework in adolescents with ADHD: comparing training in self- vs. parent-monitoring of homework and study skills completion" (2005). LSU Master's Theses. 2940. | https://digitalcommons.lsu.edu/gradschool_theses/2940/ |
Opt-in vs. Opt-out = opt-in-opt-out?
On the activation of the ICC’s jurisdiction over the crime of aggression
In December 2017, the States Parties to the International Criminal Court (ICC) had the chance to realize in New York what Robert H. Jackson called for in his opening statement before the International Military Tribunal in Nuremberg. While international criminal law was only applied against German aggressors at that time, the U.S. Chief Prosecutor emphasized that the condemnation of aggressive war should be the benchmark for any other nation in the future. When states laid down the foundations of the ICC in 1998, they listed the crime of aggression as one of the crimes under the Court’s jurisdiction. Although States Parties agreed on a definition of the crime of aggression and the conditions for the Court’s exercise of jurisdiction at the 2010 Kampala Conference, it took another seven years before they activated the Court’s jurisdiction. This article sheds light on the most controversial issue prior to activation and explains the content as well as the effects of the resolution as adopted by the Assembly of States Parties (ASP).
The remaining controversy with regard to non-ratifying States Parties
States were divided on the following, very technical question: In case of state referrals or proprio motu investigations, does the ICC have jurisdiction over nationals of States Parties that have not ratified or accepted the aggression amendments and that did not make an opt-out declaration? Intuitively, international lawyers would say “no”. Treaty amendments cannot bind a state without its consent. But this does not answer the question whether active consent (by accepting/ratifying the amendments) or passive consent (by not making an opt-out declaration) is required. Several states, the so-called opt-in camp, argued that active consent by the aggressor and the victim state was necessary. Why? According to the plain meaning of the second sentence of Article 121(5) ICC Statute, the ICC shall not exercise jurisdiction regarding a crime covered by an amendment unless the state of nationality and the state of territoriality have opted in by accepting/ratifying the amendments. This sentence seems to establish an opt-in system in case a new crime is added to the Court’s jurisdiction. The opt-out camp tried to rebut these allegations. Whereas any new crime (e.g. terrorism) needs to fulfill all requirements of Article 121(5), this cannot hold true for the crime of aggression which is already listed in Article 5(1) as a crime under the Court’s jurisdiction. In addition, the application of Article 121(5) second sentence would conflict with the genesis of the Kampala compromise: To bridge the gap between those that did not require the consent of the aggressor state (Article 12(2)(a)) and others that emphasized its importance, delegates established an opt-out system (Article 15bis(4)). Accordingly, the Court can exercise jurisdiction over a crime of aggression committed by nationals of a State Party, unless the aggressor state has previously revoked its consent by opting out. Thus, States agreed on a consent-based regime, but they decided that passive consent sufficiently respects the interests of aggressor states. The opt-out regime would lose its character as a middle ground if it was combined with the opt-in clause of Article 121(5). States could shield their aggressive leaders twice, by not ratifying the amendments and by opting out after ratification.
Establishment of an opt-in-opt-out system but reaffirmation of judicial independence
In the end, it was not the power of legal arguments but the risk of an indefinite postponement of the activation decision that led to the adoption of the resolution, which is mainly based on the opt-in regime. Paragraph 2 reflects almost verbatim Article 121(5) second sentence by stating that the Court shall not exercise its jurisdiction regarding a crime of aggression when committed by a national or on the territory of a State Party that has not ratified or accepted these amendments. If one perceives the resolution as subsequent agreement or practice (Article 31(3) Vienna Convention on the Law of Treaties), the combined reading with Article 15bis(4) leads to an opt-in-opt-out system. Good news for all states that change their mind after ratification. There would be an emergency exit in case they decide to attack other countries after ratification. Bad news for the 35 states that expected judicial protection after ratification. Except for Security Council referrals, their aggression case against non-ratifying States Parties could not be brought before the ICC. End of story? Well, the resolution still leaves some leeway by reaffirming judicial independence (paragraph 3). In case the resolution remains ambiguous as to the scope of jurisdiction, it is up to the Court to decide, see Article 119(1). Indeed, it is quite contradictory to establish an opt-in regime while “noting with appreciation the Report on the facilitation” which summarizes the views of States Parties, including those supporting the opt-out camp. Nonetheless, it will be hard to justify that the pure opt-out system of Article 15bis(4) still applies despite the clear wording of paragraph 2 of the resolution. On the other hand, there is a slim chance that subsequent state conduct might reveal a persisting controversy that needs to be settled by the Court. The subtle objections States Parties expressed after adoption might serve as a first indication. States could further strengthen the impression of disagreement by declaring a temporary opt-out until they ratify the amendments. This seemingly schizophrenic conduct would help to reaffirm the pure opt-out regime as the prevailing one despite the formal adoption of the opt-in clause. Their opt-out declaration would suggest that the default rule is acceptance (Article 15(4)). If states fail to engage in common conduct, paragraph 2 of the resolution cannot be seen as establishing subsequent state practice. Be that as it may, time will tell whether the paragraph on judicial independence will ever become a powerful tool for the Court. At the ASP, however, its inclusion was rather a means to placate the opt-out camp and to reach consensus.
The potential impact of the resolution
In case paragraph 2 is perceived as a subsequent agreement, the ICC’s jurisdiction will be further restricted in case of state referrals and proprio motu investigations, namely, both the aggressor and the victim state need to ratify before the ICC can exercise jurisdiction. If the number of ratifications does not increase, these two trigger mechanisms will only play a minor role in combatting aggression. Instead, referrals by the Security Council will gain in importance. Contrary to States Parties and the Prosecutor, the Security Council can trigger the Court’s jurisdiction even if Non-States Parties or non-ratifying States Parties are involved (Article 15ter). The adopted opt-in clause does not only reduce the number of potential aggression cases from conflicts between 122 States Parties (Kenya opted out) to 35 ratifying States Parties. Indirectly, it also reduces the Court’s autonomy from the Security Council as its exercise of jurisdiction over non-ratifying States Parties would fully depend on a Security Council referral. Ironically, the permanent members already called for the Council’s exclusivity to trigger the Court’s jurisdiction in Kampala. Seven years later, States Parties did not establish exclusivity, but they adopted a resolution that potentially produces the same effect if the number of ratifications does not increase. The condemnation of aggressive war will be a benchmark for any other non-ratifying nation, but only if the Security Council so decides.
Annegret L. Hartig works as a research assistant at the University of Hamburg and writes her PhD thesis on the domestic implementations of the crime of aggression. At the 16th session of the Assembly of States Parties, she had the chance to follow the negotiations on the activation of the International Criminal Court’s jurisdiction over the crime of aggression.
Cite as: Annegret L. Hartig, “Opt-in vs. Opt-out = opt-in-opt-out?”, Völkerrechtsblog, 7 February 2018, doi: 10.17176/20180207-093321. | https://voelkerrechtsblog.org/opt-in-vs-opt-out-opt-in-opt-out/ |
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https://julkaisut.valtioneuvosto.fi/bitstream/handle/10024/162005/TEM_oppaat_10_2019_Tutkimusartikkeleita_kotoutumisesta_20012020.pdf?sequence=1&isAllowed=y
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We examine the link between voter turnout and institutional features of electoral systems such as the threshold of exclusion and proportionality, and other empirical factors such as competition and the effective number of parties, i.e. factors that previous literature has suggested will probably be closely linked to district magnitude. As in Blais...
Using survey data on Finnish local politicians (n = 364) we examine the extent to which political orientation affects attitudes towards and perceptions of competitive tendering in social and health service provision. Expectation of cost benefits turns out to be the most important factor accounting for willingness to increase the usage of competitiv...
In contrast to many other countries, the Finnish open-list proportional representation (PR) system with its mandatory preferential voting provides an opportunity to study gender-based voting empirically. Using the 2007 Finnish national election study, the article presents an analysis of the grounds for same-gender voting, including motivations rela...
Until recently, voters’ views on the representational roles of MPs have been a largely unexplored field in the studies of political behaviour. With the exception of the work by Carman, and Mendez‐Lago & Martínez, the few existing studies have mainly been conducted in the US and are fairly dated. In this study, we are partly filling this gap by exam...
Deriving from the general theories of representation, as well as from the economic interest thesis on the issue of immigration, the paper examines the opinion congruence on the work-related immigration issue between voters, non-voters, elected MP candidates and non-elected MP candidates in the Finnish parliamentary elections of 2003. The study is b...
The relationship between age and turnout has been curve-linear as electoral participation first increases with age, remains relatively stable throughout middle-age and then gradually declines as certain physical infirmities set in (see e.g. Milbrath 1965). Alongside this life-cycle effect in voting, recent pooled cross-sectional analyses (see e.g....
The study examines simultaneously the effects of age, generation and period in the Finnish parliamentary elections of 1975–2003 based on pooled data from Finnish Voter Barometers (N=8634). The probability of voting rises with age among three younger generations eligible to vote, and both during the period of increasing and stable turnout (1975–1983...
This article examines the extent to which political socialization accounts for generational differences in electoral participation found in recent studies. Political socialization is defined as the learning process in which an individual adopts various political attitudes, values and patterns of actions from his or her environment. The analysis is...
We examine the association of four socioeconomic factors with turnout in Finland in three age groups. The analyses are based on individual-level register data from electoral wards from the parliamentary elections of 1999 linked to population registration data on personal characteristics covering the whole 25 to 69 year-old Finnish electorate. The...
Based on individual-level register data from Denmark and Finland, we estimate the causal effect of past eligibility on turnout in two previously unexplored institutional contexts. Our dataset includes the official turnout for about 2.3 million adult Danes and about 81,000 Finns between the ages of 18 and 30. Applying the approach suggested by Mered...
As representation is based on a reciprocal relationship between a representative and his/her constituency (Castiglione and Warren 2006, 11), a correspondence between citizens’ expectations and MPs’ own role perceptions is particularly relevant. In this study we contribute to a growing field of research by comparing MPs’, non-elected candidate’s and...
Projects
Projects (6)
Research project 2019-2021 at the University of Helsinki, funded by the Finnish Cultural Foundation FACE examines Finnish emigrants' political participation, especially voting. A survey conducted with a sample of 10.000 Finnish citizens permanently living abroad constitutes the main part of the project. The survey is currently being prepared and the invitations to participate will be mailed to the prospective participants at the end of April.
BIBU explores how globalization changes citizens’ political capacities, interests, and emotions, and how the political system responds to these changes. My role in the project is in the "Democracy incubator", where we develop democratic innovations. In particular, we are evaluating participatory budgeting processes at the City of Helsinki and some other municipalities. | https://www.researchgate.net/profile/Hanna-Wass |
Tech giants to face MPs’ questions over coronavirus disinformation
Tech giants Facebook, Google and Twitter are to face questions from MPs over the spread of coronavirus disinformation.
Senior executives from the three companies are to appear before the Digital, Culture, Media and Sport (DCMS) Committee next Thursday.
Social media and internet platforms have come under scrutiny over the ability of disinformation and false claims relating to Covid-19 to spread online.
The committee said it will ask the companies about the measures put in place to stop the spread of false narratives, including conspiracy theories around 5G being linked to the outbreak, and false Twitter accounts posing as NHS hospitals and staff.
The companies will be asked how they plan to slow the “infodemic” of false claims appearing online.
Earlier this week, Culture Secretary Oliver Dowden told the committee that the Cabinet Office was rebutting around 70 false narratives about the pandemic every week, as part of a cross-Government disinformation unit.
Mr Dowden praised social media companies for “how they have stepped up to the plate”, but said the DCMS had spoken to them about how they could “beef up” their systems, as well as addressing the speed of action and working out of hours.
“The biggest thing that struck me is just the need for speed of response,” he said.
“If misinformation is out there, you know it’s a bit of a cliche, but it can be round the world in moments, so the need to not sort of say, ‘we’ll look into this and come back to it’ … we need fast action to nip this kind of stuff in the bud.”
Social media and internet firms have taken some steps to try and curb the spread of disinformation – Facebook, Google and Twitter all direct users to official health guidance on Covid-19, while on Wednesday Twitter said it would start removing more posts on “unverified claims” around the pandemic.
The new action would include any posts which encouraged people to take action against 5G masts, with Twitter confirming it would be focusing on accounts inciting the public to engage in harmful activity which “could lead to the destruction or damage of critical 5G infrastructure, or could lead to widespread panic, social unrest, or large-scale disorder”.
It comes amid a spate of mast vandalism attacks across the country, driven by unproven conspiracy theories linking 5G to coronavirus shared on social media.
Facebook-owned WhatsApp also recently announced a limiting of its message forwarding feature in an effort to make it harder for disinformation to spread on the platform. | https://www.aol.co.uk/news/2020/04/24/tech-giants-to-face-mpsa-questions-over-coronavirus-disinforma/ |
D.A.P.S. was asked to investigate a personal residence in Townsend, Delaware that was built around 1924-30. This house is nearly 100 years old and had been in the same family since it was built. It is still owned by the same family, however it has not been occupied for several years. One of the reasons the house sits vacant is the report of paranormal activity at the location.
D.A.P.S. was called in by the current owners to investigate strange noises and activity they have experienced. We were given a tour of the home, and told about the activity they have experienced there. The owner and family members claimed experiencing objects coming off walls, and apparitions moving about, as well as hearing voices. D.A.P.S. was told of bangs and objects being manipulated in the various rooms particularly the attic. D.A.P.S. was also told of hearing walking, shuffling and movement of objects in the attic.
This case has been one of the more difficult cases for D.A.P.S. We have many stories and claims by family members of paranormal activity. This we take into consideration seriously when we do our investigation. During the first investigation of this location, D.A.P.S. spent nearly 4 hours investigating the claims put forth by the homeowners and family members. After the collected material was reviewed however, the result was inconclusive. In all of the material reviewed we got one photo with a possible anomaly in it, out of nearly 150 photos. We found no evidence of apparitions, or object movement in any of the videos. We found no voice EVP recordings, and we had no exceptional personal experiences. This was leading the team to conclude that if there were a haunting at this location that it was not residual. A residual haunting is one where an event occurs in a predictable pattern such as doors opening or footsteps at the same time and location each day. This type of evidence was not found.
We also found no evidence to support the idea of an intelligent haunting. This is where spirits will attempt to interact with investigators. Voices will be captured, intelligent responses of some sort will be captured to questions, and on occasion object movement will occur. We captured no such evidence. We also did not experience or capture any evidence indicating a Demonic or evil entity was haunting the location in spite of several reports by one family member to this possibility. Demonic hauntings will show evidence of object movement, linear placement of objects such as bottles lined up in a row in a hallway, or all kitchen cabinets being opened. D.A.P.S. captured no such evidence. A cross was placed in the back room and it was never moved outside of it being done by human hand. Invoking the name of God and praying were also tried to stir up a reaction from a Demonic entity and none were forthcoming.
This lead us to the possibility of the existence of a negative entity. These are not as malicious as a Demonic entity, but are mischievous and will torment the occupants of a dwelling. With the recording of as yet unexplained noises in the attic and some seemingly valid responses to questions in two instances, it was decided that we would ask the home owner if we could return and do a session during the witching hours as suggested by one family member who noted that most activity always took place late at night. Permission was graciously granted to do a late night investigation to finally get the answers we seek regarding what may be happening at this location. The evidence we captured will indeed produce answers.
With this information, the D.A.P.S. team set up our equipment and paired off into teams of two. D.A.P.S. investigated the home for approximately 7 hours during two different investigations using all available equipment and techniques. At some points during the investigations, family members who were the victims of alleged spirit activity were welcomed to join the team as they investigated. This is commonly known as presenting a trigger object. This would be a person or thing which seems to attract paranormal activity. One of the important tasks for a paranormal group to do is to capture solid evidence of paranormal activity, and to capture evidence that backs up what the owner claims is happening at the location. It's also important to capture data that will back up experiences had by the paranormal investigators during their investigation. However, its also important to capture evidence that explains an activity as not being paranormal, but of earthly origins. Upon review of the evidence, we did find what we think is solid proof and answers as to what may be happening at the residence.
We will cover Video evidence first, followed by Photo and EVP clips. One note regarding Orbs. Many paranormal professionals dismiss Orbs outright. Far too many groups try to present paranormal Orb "evidence" that allegedly show Orbs when in fact they are bugs or more likely dust. D.A.P.S. does not completely dismiss Orbs as a legitimate paranormal event, however, D.A.P.S. is very cautious about presenting what we believe may be legitimate Orbs unless there is compelling evidence as to the Orb being a legitimate paranormal event. In the following Video, D.A.P.S. has captured what we believe are possible legitimate Orbs. Our reason for believing so is the fact that D.A.P.S. captured no other Orb's in any of its photos of other locations in the home, and what appears to be their interference with D.A.P.S. equipment. Note that this is not conclusive evidence of spirit activity, and the camera interference may be a total coincidence.
Our findings are as follows.
Video Evidence
One of the claims by the family was hearing noises and shuffling coming from the attic. The DAPS team caught what we believe is two possible paranormal ORB’s in the attic as shown in the video on the next page. A few minutes after we started recording during the first investigation, Cam 1 which was in the attic experienced what looks like a slight power drain making the focus blur. Moments later two identical ORB’s pass over the camera and in tandem make their way in a swooshing pattern up into the ceiling. Note the extreme brightness of the Orb’s and their perfect round shape. We believe that this is possible legitimate paranormal Orb activity.
Note these Orb's are the only bright circles of light seen, and move in tandem.
Photographic Evidence
One area where activity was reported was the back room where the owners reported seeing shadow people, or black apparitions going up or down the walls. While investigators David and Kim were doing a sweep during the first investigation, they snapped the photo below showing a possible white mist in the area the home owner reported seeing activity. We witnessed this same phenomena in photos taken late during the second investigation and found that if the camera was cold and the investigator held the camera in a certain way that their finger fogged the lens. A photo taken during the second investigation showed the very edge of finger tip in the shot backing up this theory.
Audio Evidence
Besides the video and digital photo evidence, the D.A.P.S. teams were able to perform a lot of EVP sessions trying to make contact. On review of the evidence, we found a few interesting EVP's. During the second investigation however we were able to debunk some of what we captured during the first investigation. These will be explained in the individual sections.
Note: For the best listening experience, use headphones and raise the WAV volume on your PC.
One of the first EVP's to be captured during the first investigation was with the digital recorder that was placed with CAM 1 in the attic. These recorders are started when the cameras are placed and left on during the entire investigation. At the time this evidence was captured, we confirmed by reviewing video that no one was in the attic or back room of the house. It was emanating from the attic as it was not picked up by the recorder in the room below indicating that the sounds did not generate from that location. What you will hear is quite clearly objects in the attic being manipulated. This happens for about 45 seconds in time. Nothing was captured on video moving so we are not sure exactly what was being manipulated. That experience is caught on this clip.
The above EVP was debunked during the second investigation. The same sounds were recorded in the attic while at the same time a mouse was captured on video moving about. Mice may not seem like big creatures, but in the middle of the night when everything is silent, and if your already suspecting something is happening in the house, a mouse can sound as big as a squirrel or cat. A family member who used to live in the house reported hearing scratching and compared it to a dog trying to dig its way down through the ceiling. This was just after they had sprinkled salt up in the attic. Salt is a barrier. It should be used to keep spirits out, not in. The salt on the hard wood floor in the attic only served to enhance the sounds of the mice scratching away up there. Investigators found clear evidence of mouse droppings in the attic as well. See below for the second EVP recording and the video of the mouse.
In the video clip provided below we show the noise maker in the attic, a mouse. We only caught this one little guy, but there are bound to be more.
Our third EVP was also caught by David and Kim. They were conducting an EVP session in the attic. David was told by family members that banging on the dry wall irritated the spirits, so he attempted to do a little provoking. David bangs 4 times and asks a question. Then he bangs again 4 times and immediately gets two bangs in response. The recording here only has his second banging and the 2 knocks response comes very fast along with one investigators footstep. It can be heard in this clip. This is later debunked.
(NOTE: This clip is a little loud as it is from the recorder David was holding so it was close to him. Lower your volume to half or less for the best audio. )
The above EVP was tentatively debunked. D.A.P.S. took the audio clip and cleaned it up and removed some background noise. It is still difficult to hear due to the volume of the recording, but we have come to the conclusion that what seems to be two knocks in response are another investigators footsteps, the later of which are a bit clearer in the clip.
Personal Experiences
D.A.P.S. teams had a few personal experiences during both investigations. Personal experiences are paranormal events that take place that affected the investigators during their investigation, but were not captured by the equipment. We will list them here as they belong with the evidence we did capture.
One personal experience was with David and Kim while doing an EVP session in the attic during the first investigation. Kim felt a scratch on her lower back. David checked her skin and saw no marks at the time. None the less its a personal experience of note.
D.A.P.S. team members also experienced several temperature drops, most commonly in the back room where a noticeable 2 to 3 degree temperature change was noted multiple times, and verified with the digital thermometer. This room always seemed to be cold and later in the investigations the temperature outside got very cold. This room also had a slight draft when the laundry room door was open as it was a non-insulated room.
Another personal experience was with Kris and Chris where they experienced a series of high K-2 spikes in the attic while they were investigating with Rob during the first investigation. Unfortunately the video Cam was too far away to capture the spike at the time. There will be many more K2 hits during the second investigation, and they will be explained later.
Another personal experience was while Kim and David were doing an EVP session in the attic. Kim was provoking by banging on the ceiling and the K2 provided 2 very low spikes. These were not captured by a camera and were weak, but its important to note that the house has no power what so ever, so it begs the question of where did these energy spikes come from? (We now have this answer!!)
The Mysterious K2 Hits Explained
The K2 meter is a digital device used to measure magnetic fields. It is commonly believed that when spirits attempt to manifest they draw energy from surroundings and this causes an energy field which can be detected. The K2 meter is a very popular device, and is used by many paranormal groups. It is however, well known that the K2 is susceptible to interference by Radio Frequencies. These are typically created by Walki-Talkie Radios. D.A.P.S. does not use their Walki Talkie Radios unless the location is unusually large. We did not bring them with us for use during either part of this investigation. What we did not know at the time was that the newer 4G cell phones also cause a Radio Frequency burst that interferes with the K2. During the first investigation we experienced a few hits but nothing major. This still had us wondering what could be causing the spikes. I had left my Cell in my truck, and none of the other team members had a 4G phone at the time, although family members were outside using various cell devices. It was during the second investigation that I had my 4G phone with me having forgotten to leave it in my truck. Again, family members and homeowners were using cell devices outside the home during the investigation. We experienced many high K2 spikes during the second investigation, that had several team members quite excited. However, when the audio files were reviewed, the Radio Frequency interference was clear as day thus debunking the K2 spikes as anything paranormal. The following EVP recordings are evidence to this fact.
Other Evidence Debunked
The second part of this investigation was quite active as far as what team members thought were personal experiences. Aside from the K2 spikes, David experienced what he thought was being touched while investigating in the attic with Brian. He was quite excited as the attic was devoid of cob webs where he was standing. However, on reviewing the video footage at that time, it appears that David had gotten close enough to the piece of plywood leaning against the wall to his left, and its course surface grabbed the hairs on his arm.
Another incident that took place was while Jenn and Val were investigating in the attic. Val claims that she was touched in the ear by something that was buzzing. A family member also made the claim that the sound of bat or bird like wing flapping could be heard in the attic. On review of the video from the time when Val experienced this event, we found that there is a large moth or similar bug in the attic. These large moths are generally very quiet when they fly around. They are however attracted to light. If the floodlight from the house across the street came on or a bright moon was out late some night, the moth will go to the source and hit the window and flap its wings and flutter trying to get to the light source. This activity during the dead of night can be quite loud and seem like a bat or bird is in the room flapping its wings. In the video clip on the next page it can clearly be seen buzzing around before it appears again and hits Val in the head.
In the video clip below a large insect believed to be a moth can be seen flying around then reappears and fly’s into the head of the investigator. At first it can be seen in back of Val’s neck then later it seems to fly from Jenn to Val and hits Val in the head and she reacts, then the insect fly’s off to the right.
Evidence Not Found
D.A.P.S. had a total of 28 hours of Video footage that was set to record in the front rooms, kitchen into bath/bedroom area, back room, and attic in an attempt to capture any object movement, shadow people, or apparitions. On review of the footage, none were detected, and during the night no evidence of the apparition or shadow person was found. We also did not capture any objects moving.
D.A.P.S. also had 29+ hours of audio that was recorded in the above locations as well as a recorder carried by the investigating team during both investigations. We heard a lot of ticks and thumps or other such noises that were caused by normal house settlement such as a floor creaking after people had been walking on it, or people moving about on other floors. Nothing that was paranormal or out of the ordinary.
D.A.P.S. also reviewed approximately 250 digital photos for these investigations, only finding one showing possible activity.
The lack of captured evidence however does not imply that the experiences explained to us are not valid. It simply means that the spirits, if present, did not choose to interact with us and our equipment during the investigation, or that those experiences can be explained as normal earthy events.
Conclusion
In conclusion, after reviewing all of the sound, photo, and video footage captured, and in consideration of the small amount of possible paranormal evidence that was found at the location, D.A.P.S. believes that there is not paranormal activity happening at the residence. The absence of any voices on any of the EVP’s and lack of interaction with investigators indicates to us that any possible haunting was not residual or intelligent in nature. Nothing in the evidence supports that. An additional investigation after the midnight hour in an attempt to capture the more blatant activity that was reported by the owners and family yielded nothing outside of normal answers to questions from the first investigation. They stated that most activity occurred after midnight, so we did the second investigation after midnight. We were told that people had to be alone, and we sent people into different rooms alone, and told them to be silent for 15 minutes, and nothing was captured. The absence of any of the indicators of a Demonic presence also excludes any activity of that nature. If the home owners would feel better about doing something more, we will supply instructions on how to perform a house cleansing and sealing. Otherwise we feel the house is devoid of spirits and spirit activity, and safe to restore and occupy.
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This version first released with ICP Printing and Graphic Arts Training Package Version 1.0.
This unit describes the skills and knowledge required to evaluate and use digital camera technology and alter and upload digital images.
It applies to individuals who may work in a team environment with some responsibility for producing quality outcomes based on requirements.
No licensing, legislative or certification requirements apply to this unit at the time of publication.
Digital media technologies
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ELEMENT
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PERFORMANCE CRITERIA
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Elements describe the essential outcomes.
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Performance criteria describe the performance needed to demonstrate achievement of the element.
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1 Assess digital camera qualities
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1.1 Camera software compatibility with hardware system is assessed and appropriate software is selected for job
1.2 Pixel resolution of camera is matched to required quality and resolution of outcome
1.3 RAM capacity of camera is checked to be sufficient for number of images required to be captured
1.4 Shutter speed, focal lengths and camera feature modes are assessed as suitable for quality and use of photographic images required
1.5 Lithium batteries are handled and stored according to work health and safety (WHS) requirements
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2 Set up for image capture
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2.1 Camera is set up for image composition according to job specifications
2.2 Lighting is arranged according to job specifications
2.3 Light intensity is set for correct exposure
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3 Preview image
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3.1 Tone curves are adjusted according to job specifications
3.2 Neutral balance of image is arranged and adjusted
3.3 Adjustments to image composition and exposure are made
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4 Photograph and upload digital image
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4.1 Digital camera is loaded and operated according to manufacturer’s and job specifications appropriate to quality of image to be photographed
4.2 File is uploaded on to relevant computer and image saved on hard disk
4.3 Photographic image files are created and stored on computer
4.4 Photographic images are enhanced, cropped and altered electronically to deliver required image
4.5 Photographic images are checked to ensure they meet job brief
4.6 Photographic images are delivered using required delivery mode
This section describes language, literacy, numeracy and employment skills incorporated in the performance criteria that are required for competent performance.
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Skill
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Performance Criteria
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Description
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Reading
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1.5, 2.1, 2.2, 3.1, 4.1, 4.5
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Writing
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4.3
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Numeracy
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1.2-1.4, 2.3
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Navigate the world of work
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1.5
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Get the work done
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1.1-1.4, 2.1-2.3, 3.1-3.3, 4.1-4.6
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Code and title
current version
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Code and title
previous version
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Comments
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Equivalence status
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ICPDMT321 Capture a digital image
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ICPMM321C Capture a digital image
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Updated to meet Standards for Training Packages
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Equivalent unit
Companion Volume implementation guides are found in VETNet - https://vetnet.gov.au/Pages/TrainingDocs.aspx?q=a74b7a0f-a253-47e3-8be0-5d426e24131d
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Release
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Comments
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Release 1
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This version first released with ICP Printing and Graphic Arts Training Package Version 1.0.
Evidence of the ability to:
Note: If a specific volume or frequency is not stated, then evidence must be provided at least once.
To complete the unit requirements safely and effectively, the individual must:
Gather evidence to demonstrate consistent performance in conditions that are safe and replicate the workplace. Noise levels, production flow, interruptions and time variances must be typical of those experienced in the digital media technologies field of work and include access to:
Assessors must satisfy NVR/AQTF assessor requirements. | https://training.gov.au/Training/Details/ICPDMT321 |
Frailty is well recognized as a geriatric syndrome that has significant association with outcomes after surgical intervention.1 Despite the recognition of frailty, there is no consensus on how to best measure or intervene on the modifiable risks that compose the frailty syndrome.2 In addition, further work is needed on how best to incorporate the finding of frailty into the decision-making process for older adults considering surgical intervention.
Identify all potential conflicts of interest that might be relevant to your comment.
Conflicts of interest comprise financial interests, activities, and relationships within the past 3 years including but not limited to employment, affiliation, grants or funding, consultancies, honoraria or payment, speaker's bureaus, stock ownership or options, expert testimony, royalties, donation of medical equipment, or patents planned, pending, or issued.
Err on the side of full disclosure.
If you have no conflicts of interest, check "No potential conflicts of interest" in the box below. The information will be posted with your response.
Not all submitted comments are published. Please see our commenting policy for details.
Shellito A, Russell MM, Rosenthal RA. Looking Beyond Mortality Among Older Adults Who Are Frail and Considering Surgical Intervention. JAMA Surg. 2020;155(1):e194638. doi:10.1001/jamasurg.2019.4638
© 2021
Customize your JAMA Network experience by selecting one or more topics from the list below.
Create a personal account or sign in to: | https://jamanetwork.com/journals/jamasurgery/article-abstract/2755268 |
Superlight: Rethinking How Our Homes Impact the Earth
One of the most powerful design philosophies of recent years has been architect Glenn Murcutt's dictum that buildings should "touch the earth lightly." Ever since the Industrial Revolution, architects have sought to liberate our houses from their solid foundations, but now climate change, new materials and restricted land use have given fresh impetus to finding lightweight solutions for our dwellings. The projects here combine two strands of thinking: that buildings can weigh less and have minimal impact on their environments, and that this lightness--visual, material, ecological--can create beautiful, ethereal houses that offer new, natural modes of habitation and greater communion with our surroundings. Each of the 40 houses selected by Phyllis Richardson--author of the widely successful XS series and Nano House--is presented through photographs, plans and lucid explanations. Residences that float on air or water, ingenious constructions using local materials, innovative structures, inflatable spaces, high-tech hyper-intelligent houses--"superlight" takes many forms. From the desert landscape of Arizona to the urban jungle of Tokyo, from rural China to mountainous Chile, this book brings new solutions for architects and designers everywhere.
DETAILS: | https://store.aia.org/products/superlight-rethinking-how-our-homes-impact-the-earth |
NEW!
Becoming Virginia Woolf: Her Early Diaries and the Diaries She Read
by Barbara Lounsberry
“They fascinate us as they fascinate her: those writers who encouraged, warned, comforted, and trained a developing genius.”—Nancy Price, author of Sleeping with the Enemy
“Lounsberry’s deeply researched and gracefully written book shows not only Woolf’s development into a great diarist but also her evolvement into the fiction and nonfiction writer revered today.”—Gay Talese, author of A Writer’s Life
“A must-read for devotees of Virginia Woolf.”—Panthea Reid, author of Art and Affection: A Life of Virginia Woolf
Starting with fourteen-year-old Virginia Woolf’s first palm-sized leather diary, this book traces Woolf’s development as a writer through the fascinating experimental stage of her first twelve diaries. Follow Woolf’s growth as she is influenced by the diaries of legendary writers like Sir Walter Scott and Fanny Burney on her way to becoming one of literature’s most renowned modernists. | https://floridapress.blog/2014/08/08/becoming-virginia-woolf/ |
The population growth of a region and its adjacent areas will create new needs. These needs are created by the advancement of culture and the increasing complexity of population activity. The emerging needs give rise to the consequent increase in infrastructure provision. The availability of adequate municipal infrastructure is believed to be one of the factors driving the city’s development as reflected in the hierarchy of the city. Central Java province has 35 districts adjacent and interact with each other. Each of these districts has a population that changes with time. City residents are considered as a society that has high economic activity that will spur revenue growth region. The development of urban infrastructure is likely to result in the development of the urban population which ultimately increases the hierarchy of the city. This study intends to see whether infrastructure developments in districts in Central Java will result in an increase in urban population in the region which further impacts on changes in the hierarchy of cities. The results show that population growth has resulted in increased population activity, transportation and technology systems along with the times. In the long run, this leads to a change of urban hierarchy. This is also because the adjacent districts will be connected and influenced. Changes in the hierarchy of cities that occurred in the years 1990-2010 in the district in Central Java province showed that the highest hierarchy position occupied by Semarang City. This is driven by the rapid development of infrastructure and technology in Semarang City. The rapid development of infrastructure and technology of Semarang City become a big enough attraction for residents of the surrounding area to move to Semarang City. | http://repository.unika.ac.id/15862/ |
Q:
A problem with truth tables
I want to know how to justify the following fact:
For the truth value of $(p \veebar q)\rightarrow p $, isn't possible to deternine $V(q)$, that's to say, the truth value of $q$. I tried to justify it by using truth tables, but I don't know how to justify this properly.
A:
The general technique for demonstrating that information can't be retrieved is to provide two examples in which the provided data is the same, but the information in question is different. For example, $q$ can't be retrieved from $p \vee q$, because we have the two scenarios $V(p) = T$, $V(q) = T$ and $V(p) = T$, $V(q) = F$. In each scenario, $p \vee q$ is true; but $q$ has a different truth value in each case. You can use a similar approach for $(p \vee q) \to p$.
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The government of Uzbekistan has finalised a public-private partnership agreement for a new thermal power plant in the Syrdarya region, which will help modernise the country’s power sector, reduce costs, and cut carbon emissions.
Uzbekistan’s JSC National Electric Grid and Ministry of Investments and Foreign Trade signed the 25-year project agreement with a consortium comprising Électricité de France (EDF), Nebras Power Sojitz Corporation and Kyuden International.
The consortium will be responsible for building and operating the 1,600-megawatt combined cycle gas-turbine power plant, which is expected to be one of the most efficient thermal power plants in the country.
Uzbekistan’s electricity generation is currently dominated by gas, with ageing and inefficient thermal plants near the end of their operating life.
The new plant is expected to be almost twice as efficient as older models, helping to decrease gas consumption, reduce the country’s carbon footprint, and support the integration of renewable energy.
“The project will help resolve key issues in Uzbekistan’s energy generation sector,” Deputy prime minister Sardor Umurzakov said.
“Firstly, the project will increase the stability of energy generation and supply,” he added. “Secondly, modern technology will ensure the efficient use of natural gas and water.”
He said successful implementation of the project would provide additional confidence for potential investors to invest in Uzbekistan’s energy sector.”
Separately, the European Bank for Reconstruction and Development said it would provide a new permanent $210 million trade finance facility to four Uzbek banks – Asakabank, Ipoteka Bank, NBU and UzPSB – to support private exporters and importers.
Private domestic businesses, including small and medium-sized enterprises (SMEs), in Uzbekistan will be better able to import necessary goods, services and equipment and export their products following the deal.
“The importance of trade has grown immensely in these turbulent times,” EBRD president Odile Renaud-Basso said.
“We are supporting Uzbek importers and importers by opening new permanent limits to our partner-banks, which will reach out to businesses across the country,” she added.
- George Russell
READ MORE: | https://www.asiafinancial.com/new-uzbekistan-power-plant-to-reduce-emissions |
Substrate-borne vibrational communication is a common mode of information transfer in many invertebrate groups, with vibration serving as both primary and secondary signal channels in Orthopterans. The Cook Strait giant weta, Deinacrida rugosa (Orthoptera: Anostostomatidae), is an endangered New Zealand insect whose communication system has not been previously described. After field observations of intraspecific interactions in D. rugosa provided preliminary evidence for substrate-borne vibrational communication in the species, we sought to identify the following: vibrational signal structure, the biomechanics of signal production, whether signal production is a sexually dimorphic trait, whether substrate-borne signals encode information regarding sender size, the primary social context in which vibration is utilized and finally, the function of vibrational signaling in the species. We used laser Doppler vibrometry to show that D. rugosa males produce low frequency (DF = 37.00 ± 1.63 Hz) substrate-borne vibrations through dorso-ventral tremulation. Rarely produced by females, male signals appear to target rivals while both are in the direct physical presence of a female. Tremulatory responses to playbacks were only produced by males in male-male-female trial contexts, and neither sex exhibited walking vibrotaxis to playback signals, indicating that substrate-borne vibrational signals are not likely a component of the courtship repertoire. While we found that vibrational signal structure was not closely related to signaler size, males that initiated duetting bouts held a significant advantage in contests. | https://findscholars.unh.edu/display/publication420660 |
I recently had an experience which triggered some trauma from earlier in my life. It resulted in a rather spectacular and intense display of memories connecting together and some emotional distress, which lasted for a couple of days. As a result of my years of practice, while experiencing the coming and going of thoughts, emotions and sensations, there was a part of me that was in the experience, but there was also a part of me that was able to more objectively observe the experience. The observing part knew that it would pass and that there was an opportunity here to learn more about myself and about my habitual patterns of mind.
During the couple of days when I was experiencing the strong emotions, I looked after myself. I took some long walks in nature and took plenty of rest and listened to my favourite music. I set some clear boundaries about what I needed to feel safe. Afterwards, I took some holiday time and connected with trusted family and friends.
As things began to calm down again, I was curious about what could be learned from the experience and so I practiced some mindful self-enquiry. This is a new practice to me, which emerged quite naturally in the development of a continuing professional development weekend (CPD) for a group of mindfulness teachers based in Italy and trained by our Italian franchisee, InnerSight. It is one of the key practices (along with resting and reflection) that Rob Nairn included in the insight training, but which so far we haven’t really developed.
The practice involves the kind of enquiry process that we lead for others when teaching mindfulness, but we do it as a reflection practice for ourselves, either at the end of a practice or after then end of a challenging situation. It involves questions such as:
- What thoughts did I notice?
- What emotions did I experience?
- What physical sensations did I notice?
- How are my thoughts, emotions and sensations connected?
- How did I feel about the experience?
- Do I notice any habitual patterns?
- Do I notice any resistance or lack of acceptance? If so, how does it manifest?
- Where does my mind wander to – past, future, present?
- Do I notice I am trying to figure things out during my practice? If so, how does this manifest? What would it be like if I let go of control?
- Do I notice striving? Do I notice I am trying to make something happen? If so, how does this manifest? What would it be like to allow my experience to be as it is?
There is no need to reflect on all the questions, just ones that feel relevant, although the first four questions are important for getting in touch with our direct experience.
When we reflect on these questions, we don’t actively think about them. Instead we drop them into the mind and allow them to mull, curious about any response that might emerge. Then we journal whatever emerges. The mindfulness teachers on the CPD weekend found it a very useful process to support the development of their enquiry practice when they are teaching. It gave them a felt sense of the space required in enquiry and of the power of simple open questions.
When I reflected on these questions after my experience, and journaled, it enabled to me to see quite clearly my habitual way of reacting in the face of conflict. It also showed me the habitual patterns that kick into place that I use to help me to feel safe. It clarified how damaging these habitual patterns can be for myself and also for others who are involved in the situation. I was able to get in touch with the wounded part of myself deep in the centre of these habitual patterns. It enabled me to forgive myself, and all involved, and to let go of blame. I was able to see a broader perspective of us each conditioned by myriad causes and conditions beyond our control.
I found the practice of self-enquiry very helpful and so I share it with you, in case you want to have a go and see what can be learned. I feel that it is an excellent tool for making our obstacles a part of the path. While each obstacle can be painful, it also presents a wonderful opportunity for learning.
I felt quite tender and bruised for a few weeks after and as I write this I feel the tenderness, a soreness, in my heart, but all is well and I feel my usual stable and happy self once more. This is what it is to be a courageous open hearted human being. I am determined to continue to walk my talk with courage and integrity so as to move towards wisdom and understanding what is the truth. This requires us to feel the authentic pain in life, but in my experience it is well worth the effort.
Anyway, if you would like to join me to practice some mindful self-enquiry, just after Christmas, then I am leading a practice day on 27th December. Perhaps we will have experienced some habitual patterns unfolding over the festive season that we are keen to enquire into? We will take it gently and mindfully, open to seeing what can be learned as we practice together, with kindness and curiosity. | https://www.mindfulnessassociation.net/heathers-musings/making-the-obstacles-the-path-2/ |
Archive for November, 2010
Stephen and I have just spent 3 and a half weeks in Ibiza, it flew by and now its time to move on.
We were very happy with our time in Ibiza, of course both of us winning the race at the beginning of our stay was perfect. Then staying on and training for ironman Cozumel has been faultless. The island of Ibiza had so much to offer us.
We stayed in an apartment on the edge of Ibiza town, (this is the biggest town on the island). Although it was the offseason for the tourists everything we needed was open. The clubs and pubs (which Ibiza is mainly known for were all closed, but Stephen and I did’nt mind too much about that as our training plan had not got much clubbing scheduled!!)
In Ibiza town there was a choice of 3 swimming pools and I think we rode on every road on the island. About 90% of the island is really quiet, beautiful and has perfect smooth roads for riding on.
Ibiza continued its great consistancy of brilliant races for me yesterday as Stephen and I both won the Ibiza ITU long course world series race!
Winning is not easy and for us both to win on the same day is very difficult! However we have done it before! But not for about 2 years. As much as we can “dream” about both winning on the same day, actually doing it is very difficult.
At 5am we woke up to a tropical storm. Well, actually I knew it was doing this as it kept me awake most the night as it battered the hotel. I took a look outside and over transition, bikes were blown about, trees blowing sideways and rain washing everything out. …I quickly stepped back inside!
This continued all morning but the rain eased up about half way on the bike.
Its funny, the day before the race we were talking to the race directer and he told us it never rains in Ibiza!! …well yesterday made up for that!
The usual calm sea was rough. I was’nt really looking forward to getting in it! But Stephen said I would be fine! so I just got on with it.
During the first lap I thought they would’nt send us in for another lap, ..but they did, during the 2nd lap I thought they would’nt send us in for a 3rd lap, but they did! I did’nt envy the weaker swimmers yesterday.
I came out the water with a bit of a slow time, and 3 and a half minutes down on the first girl. The bike was windy and wet. The first corner we turned was into a strong headwind and up hill, but no problem, I just got on with it. My favouride part of my day was seeing Stephen leading the bike AND looking strong and looking like he was attacking the race!
The bike course was out and back and every time I saw him he looked better and better. THAT is the Stephen I want see. I know he can race like that. So that was my favorite part of my day, seeing Stephen on every lap in the lead!
After 10 km on the bike there was 20km of roller coaster roads. It was very slippy because of the wet weather, people were struggling to stay on their bikes. Unfortunatly I saw too many crashes yesterday. I wish every one who came off a speedy recovery.
I rode well and actually started the run with a good 30 minute plus gap over the next girl. For me that was nice since I am not running like I used too! I ran the best I could but it was’nt so fast especially towards the end. My legs are not springy like they were in years gone by! But I still came accross the finish line in first place. A double Bayliss win! Yeahhhh! That is what we love to do! If feels very special and we will enjoy our win here.
A huge thankYou to Emma for supporting us ALL day, and every day! You are the best.
| |
Suzette Kroll, RD, is a well-regarded expert in the nutrition field. She is a registered dietitian, certified intrinsic coach, and specialist in adult weight management. She has been inspiring good nutrition habits in others since 1991. Suzette’s advice has been seen in Seventeen Magazine, The American College of Sports Medicine Fitness Journal, Weight Watchers, Mindful Mama Magazine, and on various cable and television stations, including The Food Network. Leading by example, Suzette is both motivational and inspirational. She is best known for her personable style, quick wit, and ability to translate the latest scientific findings into advice about what to eat for dinner. Her hobbies include yoga, weight training, flamenco dancing, harmonium playing, writing, and chocolate chip cookies. Not necessarily in that order. | https://www.saguarosurgical.com/physicians/suzette-kroll/ |
Are stock prices inflated?
While stock prices may be inflated above what current business fundamentals can support, 15% growth is much less than the degree of price inflation typical of a full-fledged bubble. By comparison, the Nasdaq from 1995 to 2000 jumped by about 400%. In certain sectors, however, a current bubble does seem apparent.
Is the market going to crash in 2021?
Many experts were convinced that stocks would crash late last year or during the first half of 2021, mostly due to the fact that the market has been largely overvalued for a really long time. But that didn’t happen. Here’s what we do know, though. The stock market is apt to tumble eventually.
Is the stock market overvalued?
Based on the latest S&P 500 monthly data, the market is overvalued somewhere in the range of 121% to 272%, depending on the indicator, down slightly from 120% to 272% the previous month.
Will the stock market crash in 2020?
The crash caused a short-lived bear market, and in April 2020 global stock markets re-entered a bull market, though U.S. market indices did not return to January 2020 levels until November 2020. The crash signaled the beginning of the COVID-19 recession.
What will trigger a stock market crash?
A stock market crash occurs when stock prices fall suddenly and unexpectedly. A major economic downturn, a catastrophic event, or the bursting of a long-term speculative bubble can all trigger a stock market meltdown.
What happens when the stock market bubble bursts?
If the bubble was large enough, the entire stock market can be pushed into a bear market when it bursts. In fact, when the dot-com bubble popped, it caused the market crash of 2001, where share prices tumbled regardless of sector or, in many cases, asset class.
Where should I put my money before the market crashes?
If you are a short-term investor, bank CDs and Treasury securities are a good bet. If you are investing for a longer time period, fixed or indexed annuities or even indexed universal life insurance products can provide better returns than Treasury bonds.
Will the houses go down in 2021?
The California median home price is forecasted to edge up 8.0 percent in 2021, following an 11.3 percent increase in 2020. … The average 2021 rate for a 30-year fixed-rate mortgage will be 3.0%, down from 3.1% in 2020. Housing Affordability Index is projected to be 27%, down from last year when it was 32%.
What will the Dow be in 2021?
In February 2020 – just prior to the global coronavirus (COVID-19) pandemic, the DJIA index stood at a little over 29,000 points.
…
Weekly development of the Dow Jones Industrial Average index from January 2020 to July 2021.
|Month/day/year||Index value|
|7/7/2021||34,888.79|
|6/30/2021||34,577.37|
What is Warren Buffett indicator?
The Buffett indicator (or the Buffett metric, or the Market capitalization-to-GDP ratio), is a valuation multiple used to assess how expensive or cheap the aggregate stock market is at a given point in time.
How do you know if a stock is overvalued?
A stock is thought to be overvalued when its current price doesn’t line up with its P/E ratio or earnings forecast. If a stock’s price is 50 times earnings, for instance, it’s likely to be overvalued compared to one that’s trading for 10 times earnings. Some people think the stock market is efficient.
What was the worst day in the stock market?
The worst day in the history of the index was October 19 1987, when the index value decreased by 22.61 percent. The largest single day loss in points was on May 2, 2018.
Why did the stock market crash March 2020?
March 2020 stock market crash triggered by COVID-19. Natural gas, food, healthcare, and software stocks earn high positive returns. Petroleum, real estate, entertainment, and hospitality stocks fall dramatically. Loser stocks exhibit extreme asymmetric volatility.
When was the last time the stock market crashes?
The most recent stock market crash began on March 9, 2020. Other famous stock market crashes were in 1929, 1987, 1997, 2000, 2008, 2015, and 2018. | https://smash-events.net/world-stock-markets/is-the-stock-market-inflated.html |
How many years is long-term recovery?
How many years is long-term recovery?
A person is considered in long-term recovery or remission when they have stopped or moderated their substance use and improved their quality of life for at least five years. The early stages of crisis stabilization are past, the damage drug use has inflicted is undergoing repair, and a “normal” life is being built.
How long does it take a person to overcome their addiction?
A TIME article gives scientific evidence that it takes approximately 90 days for “the brain to reset itself and shake off the immediate influence of a drug.” Researchers from Yale University found a gradual re-engaging of proper decision making and analytical functions in the brain’s prefrontal cortex after an addict …
What are the 5 phases of recovery?
The five stages of addiction recovery are precontemplation, contemplation, preparation, action and maintenance….Read on to find out more about the various stages.
- Precontemplation Stage.
- Contemplation Stage.
- Preparation Stage.
- Action Stage.
- Maintenance Stage.
Is addiction life long?
The myth of the addictive personality It depends on which model of addiction and recovery you subscribe to. If you are a traditionalist who believes that addictions last a lifetime, that people readily substitute addictions, and that people have ingrained “addictive personalities,” the answer is: absolutely not.
How do I get rid of my drug addiction?
10 Ways ANYONE Can Stop Addiction Now
- Admit There Is A Problem. The hardest part to recovery is admitting you have an addiction.
- Reflect On Your Addiction.
- Seek Professional Support.
- Appreciate The Benefits of Sobriety.
- Identify Your Triggers.
- Change Your Environment.
- Exercise.
- Accept The Past.
How long does it take to break an addiction to social media?
But, it’s best to commit to your routine and try not to break it (even if it’s just “no social media after 9 p.m.”) for at least three to four weeks, according to Jones. “From a behavioral point of view, doing something for three weeks or at least 21 days will allow you to form a new habit.
Do you ever recover from addiction?
First off, this study found that 22.3 million Americans have overcome an alcohol or other drug problem — that’s 9% of U.S. adults at the time we did the survey! That nearly 1 in 10 U.S. adults have overcome a substance use problem is testament to the fact that not only is addiction recovery possible, it’s common.
What addiction does to the brain?
After repeated drug use, the brain starts to adjust to the surges of dopamine. Neurons may begin to reduce the number of dopamine receptors or simply make less dopamine. The result is less dopamine signaling in the brain—like turning down the volume on the dopamine signal. | https://stockingisthenewplanking.com/how-many-years-is-long-term-recovery/ |
During their weekly Tuesday meeting, Jonathan and Sherri started to prepare the marketing materials to be used for the upcoming showing in March. They also discussed the idea of having up to 3 guest artists join Second Story for the event, dancing pieces choreographed by Jonathan and Lorena.
Today’s rehearsal
Prior to Mandy’s arrival, Lorena and Jonathan begin their warm up by stretching at the barre. Before long, Sebastian grabs mom’s hand to join him in raiding Steffi Nossen’s toy stash. The newly acquired cars and trucks capture Sebastian’s attention for a few minutes, allowing Lorena to return to stretching.
Mandy changes quickly after she arrives and joins her colleagues for a warm up with -- drum roll please – all 3 dancers! It’s been a while since they have been able to warm up together.
As soon as Lorena begins the portion of the warm up on the floor, Sebastian recognizes his opportunity – and takes a running leap to land on her back. Unwilling to give her up just yet, Sebastian drags Lorena to one side of the room, ostensibly to eat. But he rejects all the food Lorena has brought to the studio, becoming fussy and irritable. Although he finally settles on eating pasta, Lorena acknowledges that what Sebastian really wants is mom’s attention. She notes that the last couple of days have been difficult for Sebastian, so he doesn’t want to separate from her for any length of time.
Once again, Lorena finds a way to integrate caring for her son with working – or, in this case, stretching. She figures out how to feed pasta to Sebastian and do floor stretches – simultaneously!
Meanwhile, Jonathan and Mandy move into a warm up segment that each one alternately leads. Once Sebastian seems more engaged in independent play, Lorena is able to join them. They all decide to do a quick across-the-floor segment, led by Jonathan. It is wonderful to see their big smiles since it feels so good -- and is so much fun – to move this way!
Alas, there is only 20 minutes left before Jonathan has to leave, so the dancers agree to begin rehearsal with a run-through of Jonathan’s piece. To prevent Sebastian from interrupting Lorena and Mandy during this period, Jonathan becomes the consummate entertainer for Sebastian, eliciting squeals of delight. Each time Sebastian runs toward his mother, Jonathan finds a way to entice him back to their play. Other than a couple of spots in which they are uncertain of the exact movements, Lorena and Mandy enjoy a successful run-through. Before moving on, they clarify the elements that are still unclear.
Lorena and Mandy begin to teach their parts to each other for the segment in which they switch roles. First, Mandy teaches Lorena an early gesture phrase. Next, Lorena shows Mandy her choreography while Jonathan stands nearby, offering additional coaching details. He notes the phrases that still need some clarification. It is rather challenging for each dancer to learn her partner’s part, danced to the same music she has already come to associate with her own role.
Jonathan notes that the changing of roles enables him to impart an “a-b-a’ ” structure to his dance; the ending mirrors the beginning, but includes some variation. He explains that this approach gives the piece a “retrograde feel.”
Mandy and Lorena continue teaching one another their respective roles, even after Jonathan leaves to pick up his kids at preschool. Fortunately, Sebastian is now mesmerized by a video, so the ladies can work with little interruption. Next week they hope to complete the remaining segment in which they change roles.
Mandy and Lorena move on to Lorena’s piece, marking it through from the beginning of the duet. Lorena offers a variety of notes: clarification of one change of position…intention to make the repeating section of alternating positions less “bumpy”…clarification of the timing of the tricky phrase following this repeating section…altering the timing at the end of the duet.
Lorena and Mandy experiment with the final phrase as Lorena would like to simplify the choreography. Once they are reasonably satisfied with their choices, the ladies run through the duet again, incorporating the changes and modifications they just rehearsed.
Mandy then walks through the solo that follows the duet in Lorena’s piece. Although she has not practiced this segment in a long time, the choreography comes back to her easily. She is only unclear about the steps constituting the final phrase of the solo. The ladies will check a video next week to clarify those movements. Lorena advises Mandy to execute the solo so it has a “light, fresh, soft, and easy” feeling. In fact, Lorena believes that Mandy does not need to dance it with much more effort than she just exerted.
It is now time to work on Mandy’s piece as she would like to include it in our upcoming showing. First, she and Lorena clarify the steps and timing of the existing choreography. Since she has not yet selected music to accompany her piece, Mandy puts on music from her iPhone so they don’t have to dance in silence. After stepping back to observe Lorena dance the piece, Mandy offers notes about how she would like Lorena to approach her work: think more linear…make shapes in space… go for bigger, wider movements and deeper pliés.
Sherri notes that Mandy seems pleased that she has the opportunity to create a dance that reflects her idiosyncratic style. Mandy makes a distinction between her style and that of Jonathan and Lorena: she feels that her approach is less about emotional expression and telling stories, and more about making shapes in space and playing with movement. Conceived as separate from the music, her choreography tends to be more linear and abstract. In fact, Lorena and Mandy come up with labels that seem to capture what is central to each dancer’s choreographic approach: Mandy=Motion; Jonathan=Emotion; Lorena=Music.
Since Mandy is not yet clear whether her piece will be a solo or duet and has not yet decided upon an approach to music, she and Lorena brainstorm about possible choices. Perhaps she’ll decide to use atonal, avant-garde music…or solo cello or sax. Lorena points out that choosing to create a duet means that you are constructing space as well as shape within that space -- and figuring out floor patterns. Mandy is inclined to have 2 dancers performing different choreography in different areas of the stage, at times coming in and out of a shared space.
No matter what decisions Mandy ultimately makes, she is enthusiastically assuming the new role of choreographer!
Leave a Reply. | https://www.processinrealtime.com/second-story-dance-project/second-story-rehearsal-55 |
Miguel Alberto Gomez argues that even though states are increasingly employing cyber operations as coercive instruments of foreign policy, such operations have had limited success. So what accounts for the failure of such operations? To help provide an answer, our author here explores how the structure of cyberspace affects coercive behavior and the attributes required for successful coercion, which include 1) the clear communication of a threat; 2) suitable cost-benefit calculations; 3) the credibility of the coercer; and 4) reassurances from a coercer that a threat will be rescinded upon compliance.
Theme
Cyberspace is a new domain for coercive operations in support of foreign policy and security with advantages for offensive actions and hindrances to its success.
Summary
This ARI provides an overview of factors crucial in our understanding of coercive cyber operations as the exercise of power through cyberspace in order to coerce an adversary into a particular course of action. It its focused on the compellent actions of the state actors though they, and non-state actors, may carry out deterrent actions as well. The first section presents the fundamentals of coercion. The second frames coercion in the context of cyberspace and surfaces the characteristics of the domain that enables it. Finally, the third establishes the causes behind coercive failure and, inversely, success. | https://swissintell.ch/coercion-cyberspace/ |
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Interaction Between Heavy Vehicles and Roads
Technical Paper
930001
ISSN: 0148-7191, e-ISSN: 2688-3627
Published March 01, 1993 by SAE International in United States
Annotation ability available
Sector:
Language: English
Abstract
This paper discusses road damage caused by heavy commercial vehicles.
Chapter 1 presents some important terminology and a brief historical review of road construction and vehicle-road interaction, from ancient times to the present day.
The main types of vehicle-generated road damage, and the methods that are used by pavement engineers to analyze them are discussed in Chapter 2. Attention is also given to the main features of the response of road surfaces to vehicle loads and mathematical models that have been developed to predict road response.
Chapter 3 reviews the effects on road damage of vehicle features which can be studied without consideration of vehicle dynamics. These include gross vehicle weight, axle and tire configurations, tire contact conditions and static load sharing in axle group suspensions.
The dynamic tire forces generated by heavy vehicles are examined in Chapter 4. The discussion includes their simulation and measurement, their principal characteristics, the effects of tires and suspension design on dynamic forces, and the potential benefits of using advanced suspensions for minimizing dynamic tire forces.
Chapter 5 discusses methods for estimating the effects of dynamic tire forces on road damage. The two main approaches are either to examine the statistics of the forces themselves; or to calculate the response of a pavement model to the forces, and to calculate the resulting wear using a material damage model.
The issues involved in assessing vehicles for ‘road friendliness’ are discussed in Chapter 6. Possible assessment methods include measuring strains in an instrumented pavement traversed by the vehicle, measuring dynamic tire forces, or measuring vehicle parameters such as the ‘natural frequency’ and ̟damping ratio’. Each of these measurements involves different assumptions and analysis methods for converting the results into some measure of road damage.
Chapter 7 includes a summary of the main conclusions of the paper and recommendations for tire and suspension design, road design and construction, and for vehicle regulations.
Recommended Content
Authors
Topic
CitationCebon, D., "Interaction Between Heavy Vehicles and Roads," SAE Technical Paper 930001, 1993, https://doi.org/10.4271/930001. | https://saemobilus.sae.org/content/930001/ |
Publisher: New York: The Viking Press, 1978.
ISBN: 0670698016
The Theater Essays Of Arthur Miller - 1st Edition/1st Printing
A most beautiful first edition/first printing in Fine condition in alike dust jacket, SIGNED by author and Pulitzer prize-winner Arthur Miller directly on the title page.
From the dustjacket: "Written for a variety of publications, with a variety of purposes, the twenty-three essays and three interviews collected here show with remarkable consistency the evolution of Arthur Miller's view on the theater. This book will be of value equally to the scholar, the critic, and the enlightened theatergoer"
Price: $750.00
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Commission proposal for the square at Caroli, Malmö, Sweden.
Done by Zunino Ignestam Studio
In connection with the restoring of the old area of Caroli in Malmö, Zunino Ignestam Studio were invited and participated in the competition for creating a public artwork with the specific material; light. The competition proposal was entitled “Between an old and a new town”.
The concept for “Between an old and a new town” consists of a large glass sculpture in the shape of a luminous jewel that has occurred out of the ground.
On the outside of the glass sculpture the surface is glossy and reflects the city surroundings and the light of day. On the inside the glass is frosted and the sculpture appear semi-transparent. The sculpture is built in a light stainless steel construction that stand out as a three-dimensional drawing in the city space.
Inside the sculpture the light pulsates like a slow “breath” in the evening and at night.
Very slowly a colour of light appears in the bottom, it increases in strength and intensity as it in a slow motion moves up through the jewel. As it reaches the top the light changes into a new colour and slowly fades away. After that a new breath of light begins at the bottom, a new colour begins the movement through the jewel.
A wooden bench is constructed on the ground so that it follows the shape of the jewel. The bench works as a meeting point and resting place for the inhabitants and guests that visit the square by Caroli. | http://zuninoignestam.com/proposals/between-and-old-and-new-town-the-jewel-rock-2012/ |
JHU Math Department ArXive Collaboration Analysis
JHU Math Department ArXive Collaboration Analysis
To familiarize myself with the software, I decided to create a data set and load it into Centrifuge.
I needed to find a data set that made the best use of Centrifuge’s strengths — visualizing big data to discover patterns and relationships.
I decided to use the arXiv. Instead of traditional but expensive academic journals, researchers can submit papers to the arXiv. Some, but not all, of the papers are preprints of articles due to appear in academic journals
Before I share my results, I should explain what a relationship graph is. Simply put, a relationship graph is a a way to visualize data that highlights the interconnections between data elements.
A simple relationship graph
In the relationship graph above, each circle represents an author. If there is a line between the circles, the authors collaborated on a paper. For example, the relationship graph above shows that W Wilson collaborated with Nitu Kitchloo. In turn, Nitu Kitchloo collaborated with Jack Morava.
If I was only interested in a pair of authors, it would be easy to use a SQL database to determine if they had collaborated. However, if I needed to identify interconnected groups of authors, a SQL database would be more difficult to use. I would need to complex nested joins to identify groups of collaborators. Even if I did manage to extract the data, I would still be unable to visualize the relationships.
The JHU Math Department
The above graph shows the entire math department, plus any coauthors up to 2 degrees of separation away. Authors currently on the JHU faculty are shown in yellow. Those who are not currently on the JHU faculty are shown in blue. The circles for the JHU authors are all the same size. For the non-JHU authors, the size of circle represents the number of articles. Observe that the graph has two large connected components. That is, there are two large groups of authors who are interconnected through collaboration relationships.
The largest component.
The second largest component.
The remaining author.
I should caveat that there were a few minor issues with getting the article metadata from the arXiv. Not every author participated in the arXiv’s authority controlsystem. As a result, some authors may have been confused with others with similar names. In addition, it’s important to remember that the arXiv isn’t the only venue for publication. Any collaboration outside of the arXiv isn’t shown in the graphs above.
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Kocaman, A. S.
Abad, C.
Troy, T. J.
Huh, W. T.
Modi, V.
Date2016
Journal Title
Renewable and Sustainable Energy Reviews
ISSN
1364-0321 (print)1879-0690 (online)
Publisher
Elsevier
Volume
54
Pages
688 - 703
Language
English
Type
Article
MetadataShow full item record
Please cite this item using this persistent URLhttp://hdl.handle.net/11693/21036
Abstract
The current supply for electricity generation mostly relies on fossil fuels, which are finite and pose a great threat to the environment. Therefore, energy models that involve clean and renewable energy sources are necessary to ease the concerns about the electricity generation needed to meet the projected demand. Here, we mathematically model a hybrid energy generation and allocation system where the intermittent solar generation is supported by conventional hydropower stations and diesel generation and time variability of the sources are balanced using the water stored in the reservoirs. We develop a two-stage stochastic model to capture the effect of streamflows which present significant inter-annual variability and uncertainty. Using sample case studies from India, we determine the required hydropower generation capacity and storage along with the minimal diesel usage to support 1 GWpeak solar power generation. We compare isolated systems with the connected systems (through inter-regional transmission) to see the effects of geographic diversity on the infrastructure sizing and quantify the benefits of resource-sharing. We develop the optimal sizing relationship between solar and hydropower generation capacities given realistic cost parameters and real data and examine how this relationship would differ as the contribution of diesel is reduced. We also show that if the output of the solar power stations can be controlled (i.e. spill is allowed in our setting), operating them below their maximum energy generation levels may reduce the unit cost of the system. | http://repository.bilkent.edu.tr/handle/11693/21036 |
The utility model relates to a chemical fertilizer ratio auxiliary equipment's technical field especially relates to a proportioning device is used in organic fertilizer production, and its work box height can be adjusted according to staff's height, improves its adaptability, reduces it and uses the limitation, and the organic fertilizer ratio in -process of two sets of differences, can mix comparatively fully, improve its work efficiency, and the convenient organic fertilizer capacity to leading -in two sets of differences of staff surveys, improves its reliability, including work box and four group's supports, work box left side top intercommunication has a feed arrangement, four group's supports all include piston rod, piston and pressure pipe, and the top lateral wall all is providedwith sealed uide bushing in four group's pressure chambers, all is provided with the needle valve switch on four group's piston rods, all is provided with the through -hole on four group's pistons, still include motor, transmission shaft and stirring leaf, two sets of observation holes department all is provided with transparent baffle, all is provided with the scale on two sets of transparent baffles. | |
In This Section
The Library has a dynamic collection of books, talking books, eItems, magazines, CD’s and films. Selection of these is based on community interest and on-going needs. Items are required to have an aesthetic, literary, entertainment and/or social value. When making a decision a range of factors are considered such as format, durability and language. Items are selected by a panel of librarians who are managed by the Coordinator Acquisitions & Collections and the Collections Team Leader. The ultimate responsibility for selection rests with the Libraries Manager.
When purchasing items we use suppliers who offer the most favourable economic terms and conditions and whose service is effective, efficient and timely.
Is there an item you would like us to include in the collection? We welcome your requests and are happy to purchase items of general interest that fit our selection criteria. If we are unable to purchase your request, items can often be borrowed through our inter-library loan service.
The Library’s Collection Development Guidelines respond to the Library and Council’s strategic plans and links to other library policies and guidelines. Frankston Library Service also supports the Australian Library and Information Association (ALIA) statement on free access to information and ideas in the context of a free and open society. Most collection suppliers are part of the Procurement Australia contract – Library Collections, Furniture, Equipment & Associated Requirements
Customer comment with regard to the collection is welcomed. There are a number of ways to register your feedback, such as customer feedback forms, email and letters. The Collections Team Leader will reply via phone, email or letter. | https://library.frankston.vic.gov.au/Members/The-Fine-Print/Guidelines-and-Strategies/Collection-Development-Guidelines |
Leveraging the modalities of feedback
The efficiency of Brain-Computer Interfaces (BCIs) in general and of BCI based post-stroke motor rehabilitation therapy, in particular, depends on the feedback that is provided [Grosse-Wentrup et al., 2011 ; Lotte et al., 2013]. Both visual and tactile feedback have proven efficient for healthy participants to control a BCI [Jeunet et al., 2015 ; Ramos-Murguialday et al., 2012] and to promote functional recovery post-stroke [Biasiucci et al., 2018 ; Mottaz et al., 2018].
Literature indicates that the type of feedback might have an impact on BCI efficiency in terms of performances. For example, a tactile or somatosensory feedback might be more effective than a visual feedback for post-stroke rehabilitation and for neurotypical persons in a multitasking context [Jeunet et al., 2015 ; Ono et al., 2014].
However, there are no information in the literature regarding the influence that feedback might have over long-term learning. There is also a gap in the literature regarding the potential impact of the profile of the participants, e.g., kinesthetic abilities, on the type of feedback to provide.
In this work our aim is to explore the impact of modality of feedback, as well as users’ neurophysiological and psychological characteristics on functional motor improvement post-stroke, BCI performances, learning, user experience and neurophysiological markers. To do so, we analyzed the literature and implemented accordingly a MI-BCI with realistic visual and tactile feedback. We also designed protocols for which we plan to include forty neurotypical and post-stroke patients. Each of them will take part in 10 MI-BCI training sessions during about an 1 hour each. Training sessions will differ depending on the type of feedback which is provided, i.e., either a realistic visual feedback alone or the same visual feedback and a tactile feedback in addition. | http://lea-pillette.ovh/FBmodalities.php |
This series of large-scale paintings features endangered shark species and create an environment that provokes a sense of power and helplessness. The paintings have transparent qualities and are hung from the ceiling to create an installation. This gives the art a sculptural quality, allowing viewers to see all sides of the paintings.
Artist’s Statement
I present endangered shark species as symbols of power but also as fragile and helpless creatures. Through the life size representation of the animals, the paintings emote thoughts of power and confrontation. This is contrasted by the murkiness and gruesome forms that surround the figures. My work creates a balance of expressive, nonfigurative forms with the sleek, slender bodies of the figures of the fish. This installation presents work that has functions of both sculpture and painting by encouraging the viewer to walk around the painting and view it from all angles. The paint is watered down and applied sporadically, letting the force of gravity naturally spread the water in uncontrolled directions, forming organic and fluid shapes. This alludes to flowing and swishing motions that seem to be the result of thrashing bodies and struggle.
Artist’s Biography
Andrea Siles is an illustrator and painter from Fairfax, Virginia. She is currently working to receive a Bachelor in Fine Arts degree with a concentration in Painting and Drawing from Shepherd University, WV. She expects to graduate in 2017. She has had her work featured in Shepherd University’s Sans Merci Literary Magazine. She has also exhibited in Shepherdstown and Hagerstown, including the Town Run Brewery, Phaze II Gallery, and Engine Room Art Space. She also worked for private clients, creating illustrations for books and painting large scale murals. After graduation, Andrea plans to pursue a masters degree in illustration. | http://www.sparksjournal.org/counter-currents/ |
Failure to Comply is Fatal: Recent BC Supreme Court Decision on Document Production
When it comes to demanding documents in a civil lawsuit, failure to comply with the rules is fatal. Document production in BC Supreme Court is governed by Supreme Court Civil Rule 7-1(10), 7-1(11), and 7-1(12) and by Supreme Court Family Rule 9-1(7), 9-1(8) and 9-1(9). In order to demand documents, you are firstly required to send a letter indicating which documents you are seeking and the grounds of relevance on which they are sought. The party receiving the demand has three options. Within 35 days, they must either list the newly requested documents on their list of documents, indicate why the requested documents are unavailable, or indicate why they are not relevant. Relevance is defined by the pleadings.
It is only after these steps are taken that a party may head to court on an application to force their production and the courts are not inclined to readily forgive a failure to comply with the rules. In Lit v. Hare, 2012 BCSC 1918, Mr. Justice Fitch emphasized the importance of complying with the rules:
“ Compliance with R. 9–1(7) and (8) of the SCFR (and its equivalent in the SCCR, R. 7–1(10) and (11)) is not optional and failure to observe its requirements will not readily be forgiven. I do not read Przybysz as suggesting otherwise, nor do I think that it particularly assists the applicant in this case. In fact, the case sounds a clear cautionary note about the failure to observe the requirements of this Rule. “
Recently, Madam Justice Baker overturned a master’s order for production of documents where the party requesting the documents failed to indicate the grounds of relevance on which the documents are sought. In laymen’s terms, you must indicate WHY the documents are being sought with reference to the Notice of Civil Claim or Notice of Family Claim and the corresponding responses.
The defendant requested further documents from RES and Mr. Kalafchi, pursuant to Rule 7-1(11) of the BC Supreme Court Civil Rules, B.C. Reg. 56/2019. Rule 7-1(11) requires parties seeking further documents to indicate the reason why requested documents should be disclosed. The reason given by the defendant for the request for credit and debit card statements was that it was unusual for a small company to have such a large volume of credit options available, and that the defendant was entitled to see who actually made the purchases claimed. Other than making the bald statement that the other requested documents are relevant, no actual reason for the relevancy of the other documents sought was provided by the defendant.
In Red Avacado Sales Inc. v. Yao, 2019 BCSC 996, Madam Justice Baker commented on the deficient request and the mandatory nature of compliance with the rule:
The purpose of Rule 7-1(11) is explained in Przybysz v. Crowe, 2011 BCSC 731, and the mandatory nature of the Rule is confirmed in Lindgren (Guardian ad litem of) v. Parks Canada Agency,2017 BCSC 721. In my view, the defendant did not satisfy the burden upon her to identify a reason for the disclosure of the bulk of the requested documents. Only with respect to the credit and debit card statements did the defendant articulate any reason for their production.
Similarly, there is no allegation putting into issue the use of the funds allegedly loaned by the defendant. The allegation is simply that the loans were made.
Additionally, Madam Justice Baker emphasized that the grounds of relevance must be obvious from the pleadings as filed and counsel is not permitted to “fill in the blanks” at a hearing:
While counsel are required to explain how documents are relevant, and such explanations often involve more detail that the pleadings themselves, the court must always be able to link the explanations to the pleaded allegations. It is not correct for a court to accept submissions of counsel which effectively create whole new unpleaded allegations. | https://www.richtertriallaw.com/failure-to-comply-is-fatal-recent-bc-supreme-court-decision-on-document-production/ |
A PLANNING committee has been advised to avoid refusing planning applications for “undefendable” reasons.
In 2018 Bradford Council's Regulatory and Appeals Committee went against planning officers' advice and refused plans for 102 houses and a 90 bed care home on land between Fell Lane and Braithwaite, Keighley.
Applicants Coshore Holdings appealed the decision, and earlier this year a planning inspector overturned the Council's decision.
The authority was also ordered to pay £8,294 in costs to the developer.
The inspector had said the committee was wrong to side with the many objectors to the application over their own planning experts.
Council ordered to pay over £8,000 to developer after 'unreasonable' decision to refuse housing plan
At a meeting of the committee held remotely on Thursday, member discussed the appeal decision.
John Eyles, major development manager told members that the inspector had disagreed with three of the five reasons the committee originally gave for refusal.
This included the loss of a community asset, the lack of affordable housing and highways concerns.
He said: "They felt that three reasons for refusal were undefendable. He acknowledged that planning officers found it extremely difficult to argue our case.
"The message is it is not a numbers game. One really good reason for refusal can be defended, but a decision can be weakened by an undefendable reason for refusal."
He pointed out that the cost awarded the the developer could have been much higher than the £8,294 awarded.
Members were told that there was no specific budget to deal with the costs of planning appeals, and that the money would have to come from other Council budgets. | https://www.thetelegraphandargus.co.uk/news/18544624.councillors-advised-refusing-plans-undefendable-reasons-developer-wins-appeal/ |
About ViolinThe violin, sometimes known as a fiddle, is a wooden chordophone (string instrument) in the violin family. Most violins have a hollow wooden body. It is the smallest and thus highest-pitched instrument (soprano) in the family in regular use. The violin typically has four strings, usually tuned in perfect fifths with notes G3, D4, A4, E5, and is most commonly played by drawing a bow across its strings. It can also be played by plucking the strings with the fingers (pizzicato) and, in specialized cases, by striking the strings with the wooden side of the bow (col legno). | http://sheethub.co/instrument/violin |
Storytelling and puppetry are almost magical for children. There is a tangible focus of attention when a puppet appears or when the page of a book is turned.
Children are in a unique phase of life where they can enter the realm of imagination with ease. Paint a picture, describe a scene, and suddenly they are right there with you. Puppets allow imaginative and logical thought to exist simultaneously and I believe this has enormous benefits on children’s creative development.
What is it about puppets that bring the magic of storytelling to life? When my bear puppet is moving and responding, (with each swish of fur as she nods in agreement) it is easy for children to believe she is alive. However at the same time, they know she is not. Often I am asked the question, ‘Is she real?’ I usually respond with,
‘She is a puppet, but sometimes she thinks she is real.’
Children can see she is ‘real’ in the sense that the puppet actually exists, however it is the power of their imaginations that allow them to believe she has a personality, emotions and a life force of her own. This is key to creativity. Children experience the power of their imaginations making something inanimate, come to life. I believe this can have a hugely positive flow-on effect throughout their life. If children are allowed to believe that their visions can manifest, they will feel empowered to create, whether it be putting a story down on paper, imagining a new scientific theory, or designing a new kitchen stool. As storytellers, we are fueling and empowering imaginations.
When I was little, I was forever being told that my huge creative plans were too difficult, not possible, too grand. However what my mum did not realise, was that her actions were painting the opposite picture. At every point she would be creating, designing and making, whether it be the dress she wanted for a function, the garden she designed, or the 40-meter bunting for the regatta. I constantly experienced the power of imagination and creativity becoming tangible.
Reverence is also crucial to the experience of storytelling with puppets. I treat my puppets with a similar care and respect that I would show to a real animal, stroking their foreheads softly and placing them gently on the ground. Why is this important? Reverence creates the atmosphere of respect. I am showing the children that creative time is valuable. I experience the puppets as if they were ‘real,’ and this gives the children permission to enter their imaginative world right by my side. In a world where practically and logic hold such a dominating presence, I believe there is great value in adults allowing children to fully experience their natural propensity for imaginative play. Furthermore, adults and children alike, are often swept away by the pure joy of the experience.
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5 thoughts on “Why I use puppetry in my storytelling.”
Wonderful post, Emma. Your big bear puppet is delightful. I can see why the children love it. You have given great reasons for parents and teachers to use puppetry when telling stories. As you say, it is great for the imagination and also encourages respect.
Wow Emma…some great thoughts expressed here based on your performance experiences. You are learning a lot. Your journey is one that is taking you from apprentice to master. Thank you for sharing your insights along the way. 😊
Thank you, Debbie. You are certainly right about learning. Every time I visit a school or kindergarten there is something new to learn. This is yet another reason why I love sharing my stories with children.
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The below article is Part Three of a series that began in the Winter 2015/2016 edition of the FARA Advocate.
"Drug Development Series: Drug Approvals and What Happens Next"
Once a drug has completed a successful Phase III trial, unfortunately the drug is not automatically on the shelves of the local drug store the next day. There are still several steps that need to be worked through before a doctor can prescribe the drug to patients.
First, the drug sponsor has to pull together all the data that they have and submit it to the FDA (or equivalent authorities outside of the US) and ask for approval of the drug. This is through a request for a “New Drug Application” (NDA) for a drug or a “Biologics License Application” (BLA) for a biological therapy (e.g. gene or protein therapy). These applications are typically about 100,000 pages long and contain all of the information about the drug from preclinical data, through the data from all of the clinical trials and also information about the process for making the drug, delivering the drug, quality control etc. It also includes information on what will be on the drug’s label - which is critical as it will determine who the drug will be approved for (e.g. all FA patients or perhaps a subset of FA patients like ones who show signs of cardiomyopathy, or those who are still ambulatory, depending on what population of patients the drug has been tested in). A company will often meet with the FDA several times while developing the NDA to make sure that it includes all of the information that the FDA needs in the application. Putting this together takes some time – for example, Santhera completed a successful Phase III study for idebenone in Duchenne Muscular Dystrophy in mid 2014. In June of this year they announced that they are in discussions with the FDA about an NDA, but the NDA has yet to be filed.
Once the NDA or BLA has been filed, the FDA has 60 days to complete a preliminary review to determine if they think there is enough information to continue into a full review phase. This seems slow, but they are reviewing 100,000 pages of information on something they may know little about! If they refuse the application, they will explain to the company why, and it may re-apply. Assuming the application is accepted, the FDA then has 10 months to review the information for most drugs, but 6 months in the case of priority review (a new FA drug would likely have priority review due to FA being an orphan drug for a disease with a high medical need). So, if Santhera’s DMD application was submitted today, an NDA would not be approved until November of 2016.
Once the NDA is approved, technically the company can start selling the drug immediately. However, the FDA will require additional studies – it requires surveillance of all new drugs for safety issues over time. In cases where drugs are approved on smaller trials, as would be the case in FA, they may request formal “Phase IV” studies where there is more formal investigation of side effects over a longer time, and/or in additional populations of patents that were not studied in the original trials (e.g. older patients, patients with more advanced or less advanced disease).
Even once a drug is legally able to be marketed, the company still has work to do. It needs to figure out how to get the drug to the patients – this may be easy if it is a pill that can be stored anywhere, or very complicated if the new treatment requires complex medical procedures to implement (e.g. something that needs to be infused into the brain). So, it needs to develop a supply chain to make sure that the drug can be delivered to patients. It also needs to negotiate with insurance plans (including Medicare and Medicaid) to ensure that the drug is covered by insurance, or it will likely be too expensive for most patients.
Finally, once it has figured all of this out, it needs to educate the clinicians that are treating patients about the new drug – who should be using it, at what doses, what side effects to look for etc. Not every doctor and every patient will instantly know about a new drug – which is why patients can help by following what is going on in the drug development world, so they can ask their clinicians about new developments, and whether that drug would be appropriate for them. Clinicians who treat a lot of FA patients, such as those in the Cooperative Clinical Research Network for FA are likely to be among the first to understand the details of a new drug, as they will likely have been involved in the trials, but patients being seen in smaller clinics may need to bring a new drug to their doctor’s attention. FARA will be providing information as needed when we get to this stage.
Stay tuned for next month's installment of the Drug Development Series.
← Read Part Two: "Accelerating Drug Development and Clinical Trials" | http://curefa.org/ambassadors/drug-approvals-and-what-happens-next |
The clouds will bring rain all morning in Whiteball. Rain will be present during the second half of the day in Whiteball.
Tuesday 25
8°C / 15°C
Wed. 26
8°C / 15°C
Thursday 27
7°C / 14°C
Friday 28
8°C / 15°C
Saturday 29
7°C / 15°C
Sunday 30
9°C / 15°C
Monday 31
9°C / 15°C
Show 15 day weather forecast »
A windy and showery weekend across the UK. There will be occasional sunny intervals at times but the...
A windy and rainy weekend is expected for the UK. The winds will be strong across northern parts of ...
Unsettled conditions are expected over the UK this week. There will be some showers accompanied by s... | http://www.weather-wherever.co.uk/united-kingdom/whiteball_v30276/ |
Q:
SQL sectioning/averaging based on different timetag/timestamps and user-chosen input (T-SQL)
I've got the following problem that I would like to cope with - I have a SQL dataset that I would like to section (e.g. like this one):
OldTimetag OldValue
2012-05-03 12:47:00 5
2012-05-03 13:00:00 1.3
2012-05-03 13:21:00 7
2012-05-03 14:56:00 5
2012-05-03 14:57:00 0.3
.... ....
Now, I want to section (and/or average) the data based on a user-chosen interval - into new timetags, e.g. every 15 minutes with the first timetag as starting point, i.e.:
NewTimetag NewValue
2012-05-03 12:47:00 4.507
2012-05-03 13:02:00 1.3
.... ....
The main constraint is that the value next to the timetag is always valid, until the next timetag appears. So the value of 5 at timetag 2012-05-03 12:47:00 is valid for the next 13 minutes until 13:00:00. The value for the first 15 minutes from 12:47:00 would be (13*5+2*1.3)/15 = 4.507. In the next 15 minutes, at 13:02:00 the value is simply equal to 1.3... (and so on)
I've come so long, that it is a good idea to make an "artificial table" first, to later join it with the old table. I'm generating that table by:
DECLARE @intStart datetime, @intEnd datetime
SELECT @intStart =min(OldTimetag), @intEnd = MAX(OldTimetag)
FROM OldTable
where OldTimetag between '2012-05-03 12:47:00' and '2012-05-03 14:57:00'
Declare @ArtificalTable table (NewTimeTag datetime, NewValue Float)
Declare @MinuteSlicer Int
Set @MinuteSlicer = 15
Insert @Hallo Select @intStart, null
While ( @intStart < @intEnd ) BEGIN
Insert @ArtificalTable
Select DATEADD(mi,@MinuteSlicer, @intStart), Null
Set @intStart = DATEADD(mi,@MinuteSlicer,@intStart)
If @intEnd <= DATEADD(mi,@MinuteSlicer,@intStart)
Break
End
This gives me an output like:
NewTimetag NewValue
2012-05-03 12:47:00 Null
2012-05-03 13:02:00 Null
.... ....
However, I'm having problems with the next step, how to join the tables correctly - can anyone give me a hint?
A:
One way is to determine the intervals (an interval is generated if it contains at least one timestamp), augment the time table with the next timestamp and then calculate the averages for each such interval by intersecting the intervals with the time table.
IF OBJECT_ID('tempdb..#values') IS NOT NULL DROP TABLE #values
CREATE TABLE #values (pk int identity, time datetime, value numeric(10,4))
INSERT INTO #values VALUES ('2012-05-03 12:47:00', 5)
INSERT INTO #values VALUES ('2012-05-03 13:00:00', 1.3)
INSERT INTO #values VALUES ('2012-05-03 13:21:00', 7)
INSERT INTO #values VALUES ('2012-05-03 14:56:00', 5)
INSERT INTO #values VALUES ('2012-05-03 14:57:00', 0.3)
DECLARE @timeSpanMinutes int SET @timeSpanMinutes=15
DECLARE @startTime datetime, @endTtime datetime
SELECT @startTime=MIN(time) FROM #values
SELECT @endTtime =DATEADD(MINUTE,(DATEDIFF(MINUTE,@startTime,MAX(time))
/@timeSpanMinutes+1)*@timeSpanMinutes, @startTime) FROM #values -- MAX(time) multiple
SELECT intervals.start
, SUM(value*(DATEDIFF(MINUTE -- minutes in intersection of [start,end] and [time,next]
, CASE WHEN time<start THEN start ELSE time END -- Maximum(time,start)
, CASE WHEN next<DATEADD(MINUTE,@timeSpanMinutes,intervals.start) THEN next
ELSE DATEADD(MINUTE,@timeSpanMinutes,intervals.start) END -- Minimum(next,end)
)*1.0/@timeSpanMinutes)) as average
FROM
(SELECT DISTINCT DATEADD(MINUTE, (DATEDIFF(MINUTE,@startTime,time)
/@timeSpanMinutes)*@timeSpanMinutes, @startTime) AS start
FROM #values -- round start to multiple of @timeSpanMinutes
UNION SELECT DISTINCT DATEADD(MINUTE,@timeSpanMinutes+(DATEDIFF(MINUTE,@startTime,time)
/@timeSpanMinutes)*@timeSpanMinutes, @startTime)
FROM #values -- union distinct with same as above but shifted with @timeSpanMinutes
) intervals -- intervals start time (end is calculated as start + @timeSpanMinutes)
INNER JOIN
(SELECT v.*,ISNULL((SELECT MIN(time) FROM #values WHERE time>v.time),@endTtime) as next
FROM #values v -- add next column to #values
) vals
ON vals.next>=intervals.start and vals.time<=DATEADD(MINUTE,@timeSpanMinutes,start)
WHERE intervals.start<>@endTtime
GROUP BY intervals.start
ORDER BY intervals.start
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The Home Health Consumer Assessment of Healthcare Providers and Systems (HHCAHPS) patient engagement survey impacts the bottom line of home health agencies substantially, by attracting referral sources and value-based reimbursement. It is imperative for organizations to understand the HHCAHPS survey, and work on improving each measure to maximize long-term growth and sustainability.
Assessing Medications – Question 5
Question 5 is one of seven questions included in the Home Health CAHPS survey Specific Care Issues domain, designed to assess how effectively healthcare providers discuss medicines, pain, and home safety with patients.
Specific Care Issues
This question evaluates the patient’s perception of whether home health providers requested to see the patient’s medications at the Start of Care. Optimizing performance on this question relies on an organization’s ability to establish and implement effective medication reconciliation processes when initiating care and during transitional points throughout care.
Suggestions for Improving Specific Care Issues Related to Medications
- Ensure clinicians request to see all the patient’s medications at Start of Care, Recertification, and Transfers (M2016), including over-the-counter and herbal medications.
- Reassess and discuss medications on each visit, and address any changes as needed. Homebound patients often take multiple medications and may need to be encouraged to provide additional medication they may be taking but forgot or neglected to reveal on prior visits.
- Implement teaching tools in staff training, and throughout medication reconciliation processes for structure and consistency.
In home healthcare, requesting to see the patient’s medications is a significant step in reconciling medications, and HHCAHPS performance will reflect successful implementation of visual assessment. Several examples of tools and resources to ensure constructive medication assessment and reconciliation are outlined below:
MATCH: Medication Reconciliation Toolkit
Through support from the Agency for Healthcare Quality and Research (AHRQ), and collaboration between Northwestern Memorial Hospital, Northwestern University Feinberg School of Medicine and The Joint Commission, the Medications at Transitions and Clinical Handoffs (MATCH) Toolkit for Medication Reconciliation was developed to improve patient safety and decrease instances of patient harm related to medication discrepancies. The MATCH toolkit provides a collection of resources to guide healthcare organizations through developing and optimizing medication reconciliation processes.
Medication reconciliation is defined in the toolkit using several clinical steps, highlighted below:
- On admission, obtain a complete list of medications the patient is taking. This list should be obtained from the patient (through visual assessment of medications) whenever possible. Other resources (e.g., family/caregiver/past medical records) should be consulted only when necessary.
- Review this list with the patient to verify accuracy and completeness.
- Once admission orders are written, compare admission orders with the medication history list to check for any discrepancies. Confirm that any variations are intentional changes. If any unintended discrepancies are discovered, consult the physician and document resulting modifications.
- If the patient is discharged or transferred to another care setting, the discharge orders should be reviewed and compared to the patient’s current medications list and reconciled to create the patient’s discharge medication list. The patient should be educated on any changes, and the patient’s discharge medication list must be communicated to the next provider of service to ensure continuity across the healthcare continuum.
Recommended resources from the MATCH toolkit:
- Talking Points: Justifying a Medication Reconciliation Project and Gaining Organizational Support (e.g., medication reconciliation as a patient safety issue, resource justification, and linking medication reconciliation with other initiatives, including CAHPS performance)
- Tips for Conducting a Patient Medication Interview
- Sample Letter to Leadership on Meetings Regarding Training and Implementation Strategy for Medication Reconciliation
- Sample Letter to Staff on Medication Reconciliation Educational Training Sessions
- Sample Staff Flier to Announce Rollout/Implementation of Medication Reconciliation Process
Check our blogs for the series on improving your agency’s HHCAHPS scores by systematically reviewing each of the HHCAHPS questions and sharing opportunities for improvement. | https://www.axxess.com/blog/cahps/improving-home-health-cahps-results-assessing-medications/ |
- Rescana Engineering
Enterprise Security - it's a Marathon, Not a Sprint.
Tacitus, one of the greatest historians and scholars of Ancient Rome, claimed that “the desire for safety stands against every great enterprise”. And while that is not necessarily true — maintaining a healthy dose of both security and free progress has always been a careful balancing act.
This is particularly true in the contemporary world, where a rising number of cybersecurity threats makes running even a small business increasingly difficult. The days when only large businesses with complex systems had to think about enterprise security are long gone.
Considering that — how should organizations approach enterprise security in a digital age?
The Need for Enterprise Security
Today, cybercrime is undeniably on the rise. Organizations and individuals alike are forced to contend with increasingly active and inventive cyber-attacks. And cybercriminals are focusing on a wide variety of targets — proving to be more adaptable than ever before. While small businesses have been their main targets for the past couple of years; the onset of the COVID-19 pandemic and the astronomic rise of remote work has shifted their priorities.
Companies of all sizes, as well as government organizations, have been targeted by cybercriminals. The need for quick deployment of a remote work infrastructure has left many companies with cyber security breaches — with cybercriminals waiting to exploit them around the next proverbial corner.
The answer to this issue is an all-encompassing approach to enterprise security. And this isn’t a one-time service that a business can obtain from an entity in the InfoSec industry; instead, we’re talking about the need for constant vigilance and a continuously alert informational security policy. But what does this entail in practice?
What is Enterprise Security?
The “enterprise architecture” has long been considered a set of “best practices” for the organization of businesses; more specifically, the roles and processes by which businesses use and create data.
And within that architecture, enterprise security is, as the name suggests, the modern security solution; one that integrates technology, data, and business practices to ensure the highest possible level of cyber security within an organization.
In practice, this means that enterprise security is not one thing; rather, it’s one of the ongoing processes within every successful business. And this particular process guarantees the safety of the business’ information assets, such as:
● Applications
● Networking
● Storage
● Workstations
● Servers
● Data
A successful enterprise security policy means that none of these company assets are vulnerable from a security standpoint — with an uncompromised availability, integrity, and confidentiality.
Enterprise security is a multi-faceted industry and discipline, which uses many tools and services to achieve this — from cutting-edge technology to detailed company policies. And the latter also serves as guidance on who is supposed to implement security policies within a company — while also spelling out who has what kind of access to which data assets.
The Shifting Tech Frontier of Cyber Crime
When it comes to the tech solutions required for a successful enterprise security process, the list is practically endless — or rather, it’s ever-evolving. At the end of the day, cyber security is a response to cybercrime. And unfortunately, cybercrime is constantly shifting and evolving as well; forcing organizations to seek equally innovative tech solutions to maintain their enterprise security.
Third-party risk management is one of the aspects of enterprise security that has gone over the most rapid changes in recent years — with OSINT-powered vulnerability discovery engines like Rescana turning the tables in the technological arms race against cybercriminals.
Tools like this are essential for the establishment of a trustworthy risk management program. And this is more important than ever, considering the amount of data that even traditionally analog companies are processing these days.
Mitigating risks is only possible through the swift identification of risky assets, the threats posed to those assets, and any measures that can be implemented to resolve them.
We work, live, and exist in a rapidly changing business ecosystem. Enterprises are changing, cyber-attacks are changing, and security paradigms are changing — which is why an effective enterprise security effort must be ready to adapt to new challenges as well.
From all of this, it appears that Tacitus’ views do not hold up today — security is the bedrock of a successful enterprise and not its hindrance. But what kind of continued approach is required to ensure long-term enterprise security?
A Team Effort
Considering all of this, it’s logical to ask — is cybersecurity a technological issue, a human issue, or a legal issue? The answer to that is simply: yes.
In practice, enterprise security stands at the crossroads between law, technology, and the human factor. With that in mind, the proper organizational and individual approach to enterprise security is just as paramount as using the most contemporary security technology.
Any successful company must have long-term and short-term business goals — and its enterprise security policy should be in line with those goals. Both line staff and business managers must have routine input in the security processes and technologies used to protect company data assets.
Every single staff member should be aware of the challenges, duties, and goals of the organization as a whole, and their peers — especially when it comes to security goals. That way, no one will be pulling in the wrong direction and thus waste company time and resources.
In other words, a high-quality enterprise security program is not exclusively top-down or bottom-up; it’s a hybrid, bi-directional model — one where every security stakeholder is involved and able to provide input. Higher executives can focus on strategy, security risks, and their effects on company growth — while middle-management and the lower staff echelons can discuss specific tools, tactics, and ground-level issues.
More Than Words
When stakeholders are not truly invested in enterprise security, it’s easy for a company’s security program to remain nothing more than a filled checkbox on the page of an auditor. However, while enterprise security is not strictly a top-down system — the approach from top brass still trickles down to the tactical and operational levels in the company.
In other words, if top executives invest serious energy and forethought into cybersecurity, other company stakeholders will follow suit. Conversely, if there’s only the minimum needed investment in cybersecurity training, tools, procedures, and policies — that kind of attitude will resonate with others as well, and they will perceive enterprise security as a secondary objective.
A truly effective security program requires a more substantial approach from the entire organizational structure. Cybersecurity has to be a clear overarching goal for the whole company, and a business differentiator on a strategic level. The road for achieving a sufficient level of security needs to be mapped out in the long term, and then clearly communicated to all stakeholders.
Quality Incident Response
While cybersecurity needs to permanently permeate the company culture, having a sufficient level of readiness for incident responses is important as well. Many companies make the mistake of having incident response teams purely consisting of technical staff.
Instead, the incident response should involve senior management, DevOps, R&D, PR, legal, and any other relevant departments.
Everyone involved in the security chain must be taught vigilance, without becoming overzealous — the incident alarm shouldn’t be triggered with false flags frequently. With that in mind, attack and threat simulations are an important part of any incident response initiatives. That way, there won’t be any panicked wrong moves.
Furthermore, any manager trying to improve the security status within an organization will need allies. In most cases, these will be security enthusiasts that are already present in the company; promoting them will go a long way to improving the security culture within the company.
A Positive Outlook
The current growth of the cybersecurity industry is reactionary — mostly triggered by the rise of high-profile cybercrime in the previous decade. However, looking at enterprise security as a mere tool of mitigating risk is not only needlessly negative — it’s also factually wrong.
In an age of rampant cybercrime, resorting to fear-mongering to raise enterprise security is the line of least resistance. However, any long-term security effort needs to be supported by clear data — both from your own company and similar industry examples from competitors.
The vigilance of a strong enterprise security system has a bright side as well — one that doesn’t necessarily have anything to do with risk alleviation. Remember those smart third-party risk management (TPRM) solutions we’ve mentioned, for example?
TPRM platforms like Rescana can help with company growth as well. Their smart solutions asset analyses and threat prediction can significantly reduce vendor onboarding times. Plus, a strong enterprise security structure can be a boon when it comes to customer acquisition as well. At the end of the day, no one wants to be associated with an insecure company in the digital age — and being perceived as secure is an attractive trait to have
Security Equals Competitiveness
Also, a methodical approach to enterprise security allows companies to empower mobile employees as well; something that has proven to be more important than ever in the age of COVID-19 and the meteoric rise of remote work.
This is something that’s not going to change back to the way things were after the end of the pandemic; according to most industry analyses, plenty of newly remote jobs are staying remote. And that means companies will need to use new techniques to vet mobile workers properly, develop and deploy new digital certificates, and stay up to date with the relevant technologies; all to remain competitive in a fast-changing world.
Easier Compliance
The rapid pace of technological development has left the appropriate legislative solutions lagging. However, “smart law” is slowly catching up — with data privacy regulations from different jurisdictions becoming more and more difficult to comply with simultaneously. And the future of data regulation is set to become even more complex; which is going to be a major issue for many companies going forward. On the bright side, however — that’s just one of the many reasons why a long-term enterprise security policy is an asset, rather than a burden.
Having a staff that’s used to dealing with cybersecurity concerns and internally trained experts means being better prepared for the changing landscape of compliance issues tomorrow. Data protection encryptions will evolve, and the companies that are already used to an evolving security landscape will be more competitive than those who become hopelessly outdated.
It Really Is a Marathon
In the end, it all comes down to one overarching theme — enterprise security is a long-term commitment. It’s not a process with a beginning and an end; especially in a digital world. Just like the need for a security guard in an office building — it’s always necessary if you want a stable and thriving company in the age of the Internet, there’s no finish line or final destination for enterprise security.
Even now, security is a firmly embedded part of our business culture. Considering that, it’s not a question of choosing between using enterprise security or not; rather, it’s whether you’ll have good security or bad security. And that’s no choice at all. | https://www.rescana.com/post/enterprise-security-it-s-a-marathon-not-a-sprint |
Community engagement at underground storage tank (UST) sites varies and is dependent on many factors including site location; severity of the release; impact to the drinking water supply; or impact to the local community or environment. Many UST releases involve relatively minor or no groundwater contamination and communities may choose not to be involved in cleanup decisions; however, some releases may have significant impacts on communities. Also, if the site is to be redeveloped or reused, communities may participate in planning and determining reuse options.
Except for UST systems located on Indian lands, states and territories are the primary implementers of the UST program within their boundaries. Federal UST regulations require that states and territories provide notice about corrective action plans (CAP), make site release information and decisions on the CAP available upon request, and hold public meetings if necessary. Additional public notice is required if cleanup levels are not achieved or termination of the CAP is considered.
Many states and territories engage with communities above what is required in the federal regulations.
Background
As of September 2021, there are approximately 542,000 underground storage tanks nationwide that store petroleum or hazardous substances. The greatest potential threat from a leaking UST is contamination of groundwater, the source of drinking water for approximately half of all Americans. EPA, states, territories, and tribes work together to protect human health and the environment from potential UST releases.
Community Engagement Resources
The EPA and state underground storage tank (UST) resources below may be useful when you conduct community engagement at UST sites. EPA welcomes your input and suggestions. We will periodically update these resources when we have more information.
Federal Requirements for Public Participation
- Regulations Pertaining to Underground Storage Tanks (40 CFR Part 280)
- Public Participation provisions 280.67 Subpart F - 280.60-280.67 - "Release Response and Corrective Action for UST Systems Containing Petroleum or Hazardous Substances"
- LUST Trust Fund Cooperative Agreement Guidelines
Guidelines for Tailoring Community Engagement Activities to Circumstances
- Indiana: Citizens Guide to Underground Storage Tanks (PDF) (3 pp, 135 K, About PDF)
- Maryland: Public Participation Guide Citizen Handbook (PDF) (37 pp, 1.6 MB, About PDF)
- Massachusetts:
- Fact Sheet: Vapor Intrusion & Indoor Air Contamination from Waste Sites
- Contingency Plan Fact Sheet: Public Involvement In Site Cleanup (PDF) (4 pp, 40 K, About PDF)
- Cleanup Program Fact Sheet: Opportunities for Public Involvement in Preliminary Response Actions
- Michigan: DNRE Public Involvement Handbook (PDF) (28 pp, 463 K, About PDF)
- New Jersey: | https://www.epa.gov/ust/community-engagement-and-underground-storage-tank-program |
With the arrival of summer, stepfamilies may be gearing up for visits from stepchildren. Often children living with one parent during the school year will spend time with their other parent for extended periods of time throughout the summer months. Extra complications may arise within the stepfamily due to changes in routines and schedules, but by incorporating a few strategies, summer transitions can run smoothly for all family members (Bonnell & Papernow, 2019).
1. Provide structure and routine for children
During the school year, children have routines and scheduled activities, so by making a summer schedule, including planned summer camps and events, children will have some consistency between both households. The schedule should also include pick-up and drop-off times and locations, so all parents are prepared. Scheduled pick-up and drop-off times also provide clarity for children about where they will be and when (Papernow, 2013). Communicating the summer plan with the other parent helps to keep them in the loop and provides important information about their children. Finally, it is helpful to give the children a copy of the calendar, whether it’s a hard copy or electronic copy. This will provide consistency and predictability for children as they transition between households.
2. Be consistent about rules and expectations at both houses
Parenting expectations and family rules are common areas of disagreements that can cause stress and tension among parents. Although children do not need both households to have the exact same set of rules, agreements on basics such as bedtimes, screen time, and curfews create consistency for children (Bonnell & Papernow, 2019). Keeping expectations similar at both houses not only instills good habits in children, but it also helps with the transition between households.
3. Create a living space for the children
Every child needs a space that they can call their own, especially if they are spending an extended amount of time in a different home. Creating a personal space for children, including a bed, closet, dresser, etc., helps them to deal with the transitions between households.
4. Create memories with your children
Co-parenting during the summer months can be stressful, but it provides opportunities for children to strengthen their relationship and create memories with their non-residential parent. Spending quality time with both parents provides reassurance for children and helps to strengthen and reinforce family customs and traditions (Ahrons, 2004).
The summer months often provide opportunities for children to visit their non-residential parent, and the transitions between households can be complicated. By focusing on co-parenting strategies, the summer experiences can also serve as an opportunity to make life-long memories for all family members.
References
Bonnell, K. S., & Papernow, P. L. (2019). The stepfamily handbook: From dating, to getting serious, to forming a “blended family.” CMC Publishers.
Ahrons, C. (2004). We’re still family. Harper Collins.
Papernow, P. L. (2013). Surviving and thriving in stepfamily relationships: What works and what doesn’t. Routledge. | https://livewellutah.org/2022/06/22/co-parenting-during-the-summer-tips-for-a-successful-summer/ |
Premiere: Cosimo Damiano – Consciousness
Sleep had become her enemy; when night began to fall the panic set in, a feeling she knew all too well now. But the source remained unknown, she couldn’t trace the unexplainable throes of her dreams — dreams that bordered on nightmares. Staying awake was not an option, though one she’d previously tried out of desperation, her only hope was to regain control of these unconscious moments, before she slipped too far beneath.
Rome-based producer Cosimo Damiano is making his return to Dj Inner Lakes’ Survive imprint for the third time with an 8-track LP titled ‘Hypnos’. In parallel to the Milanese imprint’s mission, Damiano seeks to connect styles and technologies from across different eras; an approach that manifests across his new album, so to does his previous work with classical and contemporary musicians. Inspired by the Greek god of sleep, he crafts his own language that examines the space between the conscious and subconscious, drawing from sonic references like acid techno producer Curley Schoop and Coil, as well as literature and cinematography.
Buy HERE. | https://www.theransomnote.com/music/premieres/premiere-cosimo-damiano-consciousness/ |
Tycho Brahe was a Danish astronomer who made the most accurate observations possible without the aid of a telescope. His detailed astronomical findings led many to question the Aristotelian notion of a perfect and unchanging universe and laid the groundwork for future scientific breakthroughs.
Brahe was born the son of a nobleman on December 14, 1546, the surviving member of a pair of twin boys. He was raised, however, by a wealthy, childless uncle and became his heir. His rank afforded him an excellent education and Brahe studied at the universities of Copenhagen and Leipzig, among others. Due to his uncle's desires Brahe entered school with the intention to prepare for a career in politics, but his own predilections led him to the field of astronomy. A discrepancy between the predicted and observed time of a partial eclipse of the sun perturbed Brahe, and he dedicated his life to making astronomy a more precise science in order to alleviate such inconsistencies. The death of his uncle in 1565 enabled him to spend more time traveling and exploring his interests. Brahe accumulated scientific instruments, began composing his own star charts, and dabbled in alchemy.
Brahe returned to Denmark in 1570, where he lived with another uncle and set up his own observatory. From the homemade post, Brahe made a discovery that would make a profound impact on his life and on astronomy. On November 11, 1572 he observed what seemed to be a bright new star near Cassiopeia and studied it for the next 18 months. Brahe was surprised to find that the star seemed to be further away than the moon and that it intensified in brightness before eventually slowly fading out of view. The event was extremely significant because it would not have been possible if the Aristotelian conception of a harmonious and unchanging universe were correct.
After publishing a brief tract regarding his discovery in 1573, Brahe became an important figure in scientific circles. In 1574, he lectured on astronomy at the University of Copenhagen before embarking on a tour of Germany, on which he interacted with a number of other prominent astronomers. Fearing that Brahe would permanently relocate to Germany, in 1576, King Frederick II of Denmark offered to provide him with the money to develop a state of the art observatory on the island of Hven, which was located near Copenhagen. Brahe accepted the proposition and construction soon began on the observatory that would be known as Uraniborg, or Castle of the Heavens. The facility, the first of its kind, attracted leading scholars from around the world.
In 1577, Brahe observed a bright comet, another phenomenon that did not correspond to the Aristotelian discipline. His measurements revealed that it, too, was more distant from the Earth than the moon, and could not, therefore, be an atmospheric occurrence as was suggested by Aristotle. Moreover, Brahe determined that the comet's path was elongate, rather than circular, which would mean that it would have to travel through the impenetrable spheres that were believed to carry the planets through the sky, a concept of Ptolemy. Brahe published his troubling findings that same year and also proposed a new theory regarding the system of the planets.
Unwilling to accept Copernicus's heliocentric theory, Brahe attempted to modify the Ptolemaic theory to coincide with his observations. In the Tychonic system, the Earth remained immobile, but the sun served as a secondary center. Mercury and Venus orbited around the sun, forming a small system that revolved around the Earth. Mars, Jupiter, and Saturn, Brahe suggested, orbited both the sun and the Earth, while the stars remained in a fixed sphere that made a full revolution each day. Although it was an interesting attempt at a compromise between two completely different viewpoints, the Tychonic system never garnered much support.
Upon the death of Frederick II, Brahe lost his financial backing and decided to relocate to Prague in 1597. There he again set up his instruments, many of which had been salvaged from his observatory at Hven, and obtained a new assistant, Johannes Kepler. When Brahe died following a brief illness in 1601, Kepler inherited all of Brahe's data and equipment. This act of bequeathal was yet another important scientific contribution made by Brahe, since Kepler would use the information to develop his system of planetary motion. | https://micro.magnet.fsu.edu/optics/timeline/people/brahe.html |
Social-Emotional and Behavioral Red Flags for Toddlers and Preschoolers
It might be hard to imagine what mental health concerns may look like for your toddler or preschooler. However, it is important to realize that children experience the same emotions as adults do. They experience happiness, sadness, anger, fear, loneliness and embarrassment, however, they do not always know how to express these feelings in appropriate ways, so it’s important to look for red flags. When their feelings get too big, children do not always have the words to use to express themselves, resulting in using challenging or unsafe behaviors to express these big feelings. These behaviors make learning, play and relationships at home, and in the classroom difficult and can be very distressing and frustrating for everyone involved.
Here is a list of common red flags that can help you to determine if your child needs support:
- Separation Anxiety:
- Extreme distress (crying, tantruming and clinging to you) when separating from you or knowing that they will be away from you.
- The symptoms last for several months versus several days
- The symptoms are excessive enough that it is impacting normal activities (school, friendships, and family relationships).
- The continuation or re-occurrence of intense anxiety upon separation after the age of 4 and through the elementary school years.
- Social Concerns:
- Little interest in playing with other children.
- Poor body awareness that impacts relationships with peers
- Failure to initiate or to participate in activities
- Difficulty making eye contact with others
- Behavioral Problems:
- Defiance: Failure to follow rules or listen to directions and is often argumentative with adults.
- Overly Aggressive Behavior:
- Temper tantrums that last more than 5 to 10 minutes.
- Excessive anger through threats, hitting, biting, and scratching others, pulling hair, slamming/throwing objects, damaging property, and hurting others.
- Difficulty with Transitions:
- Difficulty focusing and listening during transitions
- Extremely upset when having to transition from one activity to another. Before or during each transition, your child may cry excessively or have temper tantrums that last more than 5 to 10 minutes.
- Excessive Clinginess or Attention Seeking with Adults
- Excessive anxiety related to being around new and/or familiar people/situations.
- Child freezes or moves towards you by approaching you backwards, sideways or hiding behind you. Your child behaves this way in most situations and no matter how you support them, they continue to avoid interacting with others.
- Attention concerns:
- Difficulty completing tasks and following directives on a daily basis.
- Easily distracted and has difficulty concentrating or focusing on activities.
- Daily Functioning Concerns:
- Toileting: Difficulty potty training and refuses to use the toilet.
- Eating issues: Refusing to eat, avoids different textures, or has power struggles over food
- Sleeping problems: Difficulty falling asleep, refuses to go to sleep, has nightmares or wakes several times a night.
Children can exhibit concerns in the above areas off and on throughout their childhood. It is when these behaviors begin to impact peer and family relationships, cause isolation, interfere with learning and cause disruptions at home and in school that it is time to reach out for support.
Who can help?
- Licensed Clinical Social workers (LCSW),
- Licensed Clinical Professional Counselors (LCPC),
- Marriage and Family Therapists (MFT)
- Psychologists
Therapists will work with your child to help them to learn how to handle their big feelings and behavioral challenges. Therapists will use a variety of modalities during sessions including play, art, calming and self-regulation strategies, behavioral therapy, parent-child therapy, and parent education and support. They can also provide parent support and coaching to assist in diminishing the challenging behaviors at home. Often these professionals will collaborate with your child’s school and can provide additional support for your child within the school setting.
NSPT offers services in Bucktown, Evanston, Highland Park, Lincolnwood, Glenview, Lake Bluff, Des Plaines, Hinsdale and Milwaukee. If you have questions or concerns about your child, we would love to help! Give us a call at (877) 486-4140 and speak to one of our Family Child Advocates today! | https://www.nspt4kids.com/category/birth-3/page/2/ |
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There are two official online communities for Leukaemia Care. The first of these is hosted by Health Unlocked and is moderated by both staff and volunteers. The secondly is hosted on Facebook and admins in the group are staff at Leukaemia Care.
These are external websites to Leukaemia Care. Anybody posting to these platforms must be aware that information is then publicly accessible and could be viewed online by third parties. We cannot be held responsible for use of information from third parties.
As an issue of safety, we would always recommend that posters avoid over sharing personal information which could identify you offline.
Please note, all the messages placed on the forums or on social media are the opinion of the contributor, not of Leukaemia Care.
We would advise that only people over the age of 16 should participate in our online groups.
The aim of our online communities is to offer a safe space where people can share information and show mutual support of others. The nature of cancer is such that we ask all users to be mindful of content that is posted as we would not seek to cause alarm or distress.
Content is moderated and all users can report content which they deem as unsuitable.
Please do not use language that is suggestive, oppressive, intimidating or threatening.
The use of profanity is unacceptable.
Postings that give diagnoses or medical and treatment advice are not acceptable. Information posted by other users should never be considered a substitute for advice that can only be provided by a health professional. While a nurse may be present in our online forums, their role is one of fact checking. The online nurse is not a replacement for your own medical team.
Please do not promote the use of alternative treatments.
Messages that are critical of individuals or health professionals, specific treatments, services or treatment units will be removed. This is to ensure that others are not discouraged from getting the help they need.
Leukaemia Care admins have the right and responsibility of banning users who do not follow the rules of the online platforms.
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Leukaemia Care cannot guarantee that you will be protected from malicious internet activity and recommends that you install internet protection software which includes antivirus, privacy protection and intruder prevention and detection.
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Anyone requesting a live chat will be responded to by a member of our helpline.
Our helpline staff are trained to provide information and support to anyone affected by leukaemia– patients, family, partners and friends.
Our helpline staff are not medically qualified and a chat with us should not be intended as a substitute for professional medical guidance. We would always encourage anyone with a health concern to speak with a member of their medical team.
Occasionally our live chat service is used by individuals who are worried that they may have a leukaemia or other blood cancer although they have not been diagnosed. We always encourage anyone with concerns about their health to speak with their GP.
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All chats are confidential – please see the section on ‘Helpline Confidentiality Policy’ for further information.
This confidentiality agreement exists between all users of our helpline, email and live chat services and the helpline service as a whole.
We take confidentiality very seriously and aim to create a trusting relationship between service users and helpline staff.
Personal details are not shared with third parties unless consent has been given, and only on a ‘need to know’ basis.
Exceptions to this are detailed below and are made only where there is a potential risk to the individual or others and/or where required by law.
to ensure that helpline staff understand and adhere to the policy.
Caller information is recorded for the purpose of returning calls, sending out requested literature and for reviewing the levels of caller satisfaction. This information is kept on a secure electronic database. Any handwritten information is stored in locked drawers.
Callers to the helpline are free to speak to staff anonymously.
Our telephone system reveals the caller’s telephone number. If helpline staff need to return a call the helpline telephone number will be withheld. Please note that if an individual has chosen to bar anonymous or withheld numbers then we will not be able to return calls.
We do not pass on caller details unless this has been agreed with the caller. Where calls relate to other areas of the charity, we will pass on contact details. Voicemail messages or emails for specific members of staff or departments will be forwarded to the relevant person. Clear consent is not sought in these cases but is automatically presumed due to the nature of the enquiry.
Email correspondence is kept securely and electronically and will be forwarded only where necessary. For example, a fundraising query will be forwarded to the fundraising team.
All info stored with LiveChat but can be exported onto Leukaemia Care computers for training and monitoring purposes. LiveChat data is stored within the LiveChat system for 12 months later. Data exported by Leukaemia Care staff may include: Approximate geographic location, IP address, length of live chat and transcript of text. Downloaded data is only used for anonymised data to inform Leukaemia Care of the number of people using the service per month. Information on location, length of time on chat or IP address is deleted from this data.
Statistical information relating to calls is collected for managerial purposes and may be shared.
An individual is perceived as a serious and immediate risk to themselves by helpline staff. This may include being actively suicidal or self-harming.
An individual is perceived as presenting a serious and immediate risk to others.
A call, email or live chat which seems to indicate abuse of any kind to children.
A call, email of live chat which indicates possible terrorist action.
When an individual threatens helpline staff or tries to prevent the helpline being used effectively to support others.
Where there is concern for the health and safety of members of the helpline team.
Where staff believe that confidentiality needs to be broken, a decision will be taken by the information and support manager in line with helpline policy and operational procedures.
The information and support manager will ensure that all staff are sufficiently trained to fully understand the scope of this policy and how to implement it in practice.
People who use our services must take responsibility for the information they divulge.
People who use our services should ensure that they are happy with the content of this policy and raise any questions they may have with the information and support manager. | https://www.leukaemiacare.org.uk/about-us/privacy-policy/ |
Lärarstudenten som subjekt och objekt : kritiskt tänkande och disciplinering i lärarutbildning
Sammanfattning: The dissertation poses the problem of control in teacher training. Control is defined as a process in which someone wants someone else to adapt to a specific model of knowledge, to thought patterns, social relationships, and obedience. The main question is:How is control performed in teacher education and what is the relationship between control and exploration?The purpose is to expose problems in teacher training that are related to forming and controlling student teachers, vs encouraging exploration and experimentation and studying how students critical thinking is developed. Data in this dissertation were gathered from interviews with 13 students participating in teacher education. The theorization based on the empirical study is made after the presentation of the material. A hermeneutical approach was used to interpret the interviews.Results are reported in eight chapters in which the interviewees' statements are categorized according to themes. The categories are based on the authors' understanding of the statements in the material. Among other things, students report that they rarely have opportunities to discuss two or more opposing viewpoints that are presented at the same time and with equal importance. Differences between teachers are neglected and sometimes hidden, which will make critical examination more difficult. Interviewees also report that there are small possibilities to work with own experiences from their practice in schools. The education is often perceived as fragmented. Moreover, modeling as a working principle in teacher education is discussed. It can encourage critical discussions if the model is used as something to examine but if it is presented as the only one it can lead to imitative learning. However, the interviewees perceive the education as an uplifting life experience which has forced them to exceed their previous capacities. It has also played an important role in the development of their thinking.The theoretical part is focused upon seeking answers on mainly existential issues, in order to highlight the theoretical possibilities of free and critical thinking. The questions of human freedom, tradition and possibilities to break traditions are discussed. Another question is the disadvantages and advantages of taking into consideration learners previous experiences. In an interpretation of Deweys' educational philosophy, the fundamental character of reflection and its consequences for critical thinking in education is examined.The concluding discussion emphasizes the necessity for education to be open to allow and encourage students' testing of arguments. Only on the basis of what is explicit and in a variety of alternating models it is possible to make a personal choice and develop critical thinking.
Denna avhandling är EVENTUELLT nedladdningsbar som PDF. Kolla denna länk för att se om den går att ladda ner. | https://www.avhandlingar.se/avhandling/51e5a4f915/ |
This summer, the Alzheimer’s Association International Conference (AAIC) took place in Chicago, Illinois, being the largest AAIC in history! This week-long conference was an intense and inspiring experience for me, a research associate working under the mentorship of Dr. Tara Madhyastha, PhD, at the UW Integrated Brain Imaging Center. It seems the field is both growing and focusing on many emerging topics, such as the immune system, as well as classic Alzheimer’s research topics like cognition and amyloid pathology. I presented my own project on the use of neuroimaging for early Alzheimer’s disease detection (Read: Our Alzheimer's Study and the AAIC). It's truly an exciting time to be a part of this effort and while the Alzheimer’s disease research field has had many challenges and failures, the collective efforts of all these dedicated researchers makes me optimistic for the future.
Increasing Opportunity
Alzheimer’s disease is currently the 6th leading cause of death in America and this number is expected to increase with our aging population. For this reason, research to understand and treat the condition has only increased as well. To support these efforts, the National Institute of Aging (NIA) at the National Institute of Health (NIH) plans to increase funding to Alzheimer’s disease research and related projects three-fold, funding up to 25% of new/early-stage projects for the next fiscal year. According to Dr. Richard Hodes, director of the NIA, the field of Alzheimer’s research is moving toward a more collaborative and “open science” environment. As a major force in promoting this movement, the NIA plans to continue its support of a more open-source/open-data research community so that with the collection and analysis of vast amounts of data, research progress will continue to accelerate.
Early Effects of Amyloid
Amyloid, a hallmark protein of Alzheimer’s disease, can build up in the brain for 10 to 20 years before reaching disease-state levels. A major question in the Alzheimer’s field surrounds the point at which individuals begin to manifest clinical symptoms. This issue is the focus of the Dallas Lifespan Brain Study, led by Dr. Denise Park at the University of Texas. Started in 2008, the longitudinal study follows 500 healthy and cognitively normal adults ages 20 to 89, who did not show significant amyloid build up at the start of the study, indicated by normal (negative) PET scan results. This database is one of the only studies following middle-aged healthy adults as they age, and scientists plan to use this resource to track the relationship between of amyloid deposition and cognitive decline before the onset of dementia.
At 4 years into this study on cognitively normal adults, Dr. Park and her team have found that the amount of amyloid in the brain correlates with rate of cognitive decline in older adults with a positive amyloid scan (a high burden of amyloid) over a 4-year time span. Strikingly, this relationship was detectable even in the middle-age cohort - however, only in scores on vocabulary tests. Early intervention will likely be key component to treating Alzheimer’s disease, and this work provides a potential window for early detection and intervention by showing that early increases in brain amyloid can indeed be correlated with subtle cognitive changes.
Network Changes in the Alzheimer’s Brain
In our brains, information is communicated through electrical signals between neurons. When we perform a particular mental task, certain groups of neurons become active in synchrony. We call this a “functional network.” AAIC plenary speaker Dr. Lennart Mucke of the Gladstone Institute of Neurological Disease discussed his work on abnormal network activity in Alzheimer’s disease. He has found that electrical activity becomes imbalanced even before clinical symptoms appear. Dr. Mucke and his group observed that this imbalanced electrical activity results in remodeling in the brain’s hippocampus and overall changes in brain network activity. Because the hippocampus is a major site involved in learning and memory function, he believes these network changes may contribute to the cognitive abnormalities in Alzheimer’s disease. In the future, Dr. Mucke says we will need to better differentiate the network dysfunction relative to the normal function in order to understand and potentially use network measures to aid in diagnosis.
Blood-Brain Barrier and the Immune System
As a highly specialized and critical organ in the body, the brain is protected from the blood by a protective structure, called the blood-brain barrier. The blood-brain barrier is highly selective in what it allows to the brain, exchanging mostly small nutrients and gas molecules and excluding large immune cells. Research over the last decade however has demonstrated immune system involvement in many neurological conditions including Alzheimer’s disease.
Dr. Michal Schwartz at the Weizmann Institute of Science, Rehovot, Israel, a world pioneer in neuroimmunology, focused much of her career on this topic; what is the immune system doing and how is it entering the brain despite blockage by the blood brain barrier. From her work, Dr. Schwartz and her lab found that immune cells enter the central nervous system through a structure in the brain called the choroid plexus, an area that functions like a gateway between cerebral spinal fluid and blood. In addition, they discovered that in the healthy brain, an immune signaling factor, IFN-g, acts on this gateway site to prepare it for important exchanges, but in the Alzheimer’s brain, this activation may be reduced. Dr. Schwartz was a major player in initially promoting the notion that the immune system may play a protective role in the brain and moving forward, she and her team are focusing on ways to harness the immune system to fight against Alzheimer’s disease. | http://depts.washington.edu/mbwc/news/article/highlights-from-the-2018-aaic |
The volume units different approaches to perceptual metaphor analysis.
The paper examines the properties of heavy as a perceptual concept, based on evidence from 11 languages. We demonstrate that the semantics of this concept is heterogeneous; lexemes of this field can be used in situations of at least three types: Lifting, Shifting and Weighing. These situations are either lexicalised as separate words or they converge in a single lexeme in various combinations following certain strategies. We also argue that different metaphorical extensions correspond to different situation types; this allows us to use analysis of metaphoric shifts as an additional instrument to establish the semantic structure of direct meanings.
The article concerns R. Wollheim’s theory of twofoldness, the one of the most acclaimed analytical theories of pictorial representation. Wohhleim has pointed to the structural distinction of seeing pictures from seeing objects and situations face-to-face, thus proposing the theory of seeing-in and twofoldness. This conception presupposes the split, simultaneous seeing of the picture’s object and material surface. Considering the basic influences, features and shortcomings of Wollheim’s theory, author specifies the question of to what extent the scope of this theory is limited to artistic pictures. Alongside with this theory’s reconstruction and critique, the article provides its interpretation based on spatially-oriented definitions of twofoldness (B. Nanay, J. Kulvicki) coupled with the actualization of the distinction between representational content of a picture and its figurative content. The first one includes all spatial relations that can be seen in it, the second one – all that can be seen in it and described by non-abstract concepts. Wollheim reframed to apply this distinction to his notion on twofoldness. But it gives the possibility to bring out the convincing definition of twofoldness as the simultaneous visual experience of pictorial surface (i.e., some material object in viewer’s egocentric space) and of representational content (i.e., semantically active spatial relations which one sees in the picture). This interpretation may be considered a useful instrument for non-reductionist description of pictorial experience and pictures’ ontological duality, hence it is devoid of Wollheim’s psychologism while maintaining his focus on visuality. Moreover, it provides some opportunities for applying Wollheim’s theory beyond the contexts of analytical philosophy of pictures, thus connecting it to the problems of phenomenological and hermeneutic theories of image.
This paper deals with professional / non-professional degree of academic discourse by analyzing cognitive metaphor models in the English written texts produced by students with different competence in economics. It distinguishes a set of features that characterize academic and professional discourse. It also focuses on the problem of developing professional competence in economics.
Recent events in Ukraine and Russia and the subsequent incorporation of Crimea into the Russian state, with the support of some circles of inhabitants of the peninsula, have shown that the desire of people to belong to the Western part of Europe should not automatically be assumed. Discussing different perceptions of the Ukrainian-Russian war in neighbouring countries, this book offers an analysis of the conflicts and issues connected with the shifting of the border regions of Russia and Ukraine to show how ’material’ and ’psychological’ borders are never completely stable ideas. The contributors – historians, sociologists, anthropologists and political scientists from across Europe – use an interdisciplinary and comparative approach to explore the different national and transnational perceptions of a possible future role for Russia.
Plato’s interest in vision and the visual is multifaceted, and complex. Visual words and images are frequent in the dialogues along with many direct and indirect discussions of physiological, intellectual, and social vision. The increased emphasis in recent scholarship on the importance of visuality in Plato is a part of a ‘scopic turn,’ the effect of which was to ground interpretations of the history of western European philosophy and metaphysics in its entirety in certain optical premises.
This article touches upon some problems in building up a lexicon for the part of universal ontology which accounts for force interactions. We have chosen certain semantic features in the lexical description as dominant ones and conducted a small survey among native speakers of Russian to prove the results.
The paper is focused on the study of reaction of italian literature critics on the publication of the Boris Pasternak's novel "Doctor Jivago". The analysys of the book ""Doctor Jivago", Pasternak, 1958, Italy" (published in Russian language in "Reka vremen", 2012, in Moscow) is given. The papers of italian writers, critics and historians of literature, who reacted immediately upon the publication of the novel (A. Moravia, I. Calvino, F.Fortini, C. Cassola, C. Salinari ecc.) are studied and analised.
In the article the patterns of the realization of emotional utterances in dialogic and monologic speech are described. The author pays special attention to the characteristic features of the speech of a speaker feeling psychic tension and to the compositional-pragmatic peculiarities of dialogic and monologic text. | https://publications.hse.ru/en/books/210703595 |
According to the CDC, one of the top-five contributing factors of foodborne illnesses in the U.S. has been associated with the use of unsanitary equipment and utensils during food handling and preparation. Tools for cleaning a food processing facility or machinery, if not adequately managed, can become vectors of key contaminants such as micro-organisms, allergens, and extraneous matter, which may eventually jeopardize food safety and quality.
To support the argument, a 1990 UK government-funded study revealed that 47% of the cleaning equipment and tools swab-sampled at a food production site had tested positive for Listeria monocytogenes. Therefore, in order to prevent or significantly minimize cross-contamination incidents within a food plant, proper selection, storage, cleaning, care, and maintenance of cleaning tools becomes absolutely critical.
Choosing the right tool for the cleaning task
Whether a site conducts dry-cleaning, wet-cleaning, or both, tool selection is a key starting point, because cleaning by itself may enhance the spread of contaminants during removal of such soils from a surface. As illustrated in the graphic on the next page, each of those cleaning actions can pose a certain level of risk.
For instance, it wouldn’t be effective to use a soft-bristled brush to remove tough grease from equipment parts. Instead, use a stiff-or medium-bristled scrubbing brush on the part submerged in a detergent water bath to avoid aerosolizing contaminants.
Some other factors that may influence the choice of cleaning equipment are as follows:
- Don’t use stiff-bristled brushes on, say, food preparation tables constructed of sensitive stainless-steel material, as these may scratch surfaces and make them more unhygienic over time.
- Special application tools could be needed to clean hard-to-reach areas, such as the use of condensation squeegees with telescopic handles to access high walls and ceilings; angled brooms with ergonomic handles to remove debris from corners and low-lying junctions; detail-brushes to clean narrow gaps, nooks, and crannies in an equipment piece, etc.
- Drain brushes should be made of durable construction as they should be able to withstand harsh chemicals, heat and mechanical action that may be necessary during deep cleaning of uneven concrete surfaces.
Defining cleaning and storage protocols
We recommend adequately cleaning a new tool before use (but after removing any stickers), and thereafter, clean, at minimum, after every use. Just like equipment and facility areas, cleaning tools are also environmental surfaces. In general, if a brush or a scraper is used on food-contact surfaces like trays or equipment surfaces, the tools must be adequately cleaned and sanitized before and after use. If cleaning equipment such as squeegees and brooms are used on non-food contact surface like floors and walls, they should be regularly cleaned, periodically disinfected, and rinsed properly with clean water.
Tool cleaning considerations would typically involve, at the least, the following important elements:
(i) Assessing TACTER parameters required for soil removal – These are: Time required for chemicals to work on a tool surface; Action by way of mechanical agitation or scrubbing; Chemical concentration and pH; Temperature of the cleaning solution; Employees trained to do the job; and Resources such as other cleaning tools and sanitation aids.
(ii) Creating a risk-based cleaning schedule – even for the cleaning tools themselves, along with providing key details on how to clean, frequency of cleaning, verification and monitoring, and allocating the employee(s) responsible for the tasks.
(iii) Ensuring that cleaning steps are understood by employees - For a wet-cleaning operation, this may include: disassembly of equipment and dry-cleaning to remove debris; pre-rinsing surfaces with potable water from top-to-bottom; applying detergent or foam from bottom-to-top; post-rinsing with potable water; post-sanitation QC inspection; and sanitizing, assembling, and drying the equipment, followed by a pre-operational inspection.
Additionally, in order to minimize the risks of cross-contamination, the cleaned tools should be stored properly on racks or shadow boards:
- With heads down that are off-floor, and distant from other tool handles;
- In a single row so that condensate from tool above does not drip on the tool below;
- That are regularly cleaned and sanitized, as appropriate.
It is vital to regularly inspect and replace damaged tools. Worn-out bristles, a badly chipped plastic block, a broken or taped-up handle, and deep cracks are some of the tell-tale signs that it’s high time you should replace a tool.
Evaluating the priorities during COVID-19
The ongoing COVID-19 pandemic has heightened the need for enhanced hygiene and sanitation within food plants. This has been principally to prevent any possibilities of SARS-CoV-2 virus transmission between employees through touch contact surfaces, such as hand tools. However, it’s also important to note that unlike the noroviruses, this coronavirus does not lead to a foodborne illness. Nevertheless, in the interest of occupational health and safety, some of the following precautions need to be taken:
- Clean and disinfect tools before and after use, and when transferred from one employee to another. Follow manufacturer’s instructions and use disinfectant that is EPA-approved to work against the SARS-Cov-2 virus.
- If a manufacturing facility is to be re-opened after a shut-down, tools for conducting a thorough deep-clean are required.
- In addition, follow WHO/CDC guidelines of washing hands, maintaining necessary respiratory etiquette, social distancing norms, and quarantining if sick.
Manual cleaning tools still remain a necessity
Automated systems may not be able to clean parts that aren’t CIP-able. Besides, CIP systems and components (e.g. spray-balls) also may need a periodic tear-down, deep-cleaning, and verification. In such situations, manual cleaning with hand tools may be the sole practical option. Hence, the role cleaning tools play in maintaining a sanitary environment within a food processing facility cannot be underestimated. | https://www.foodengineeringmag.com/articles/99211-use-care-and-maintenance-of-cleaning-tools-during-the-pandemic-and-beyond |
How often do chickens lay eggs?
And whenever they lay eggs, have their eggs been fertilized?
2 Answers
- οικοςLv 71 month agoFavorite Answer
That depends on the breed and the season. Generally, it takes 25 hours for the ovum to pass through the oviduct and become an egg. However, there was one hen, an Australorp, that laid more frequently. However, that breed is only a moderately good egg-layer. The best layers, in general, are the white Leghorns. You also have to factor in that, when the days get shorter, hens stop laying. You can keep them going through the winter with artificial light in the mornings. Some also stop when they go broody (Silkies and Austrolorps have the reputation of being good brooders) or when they molt. Figure that you have a good egg-layer if you get 180 to 220 eggs per year. If you get more, that's the one to breed from.
The eggs will be fertilized if you have a rooster in with them. BTW, a cockerel is an immature male and cannot fertilize the eggs.
- Login to reply the answers
- .Lv 71 month ago
A laying hen will lay everyday, sometime twice. If there is no cockerel (caged birds there isn't) all her eggs will be unfertilized Anon. | https://answers.yahoo.com/question/index?qid=20200602144518AAHQ8zO |
Where shall I lay my eggs today?
The usual nesting choices. Top left: The new nest in the kindling pile (preferred by Honey, and the older hens, i.e., Lottie, Lulu or Leila); Top right: Pompom’s choice is under the edge of the barbecue cover; Bottom left: Francesca, Pearl (in situ) and Hannah use the nesting boxes in the hen house. Bottom Right: Fatima always lays in a nest box in the barn.
As much as I love our hens dearly, sometimes they can be very annoying. This particular tale concerns a couple of our ‘saved’ hens, Honey and Perky.
In mid-September, Honey went missing for almost a week. Then on two mornings in a row, Ben spotted her eating pellets in the barn with the other hens – as if she’d never been away. Then she’d disappear again.
After some sleuth work (which involved spying and following), we found her in a grassy hollow in the back paddock, sitting on a mountain of warm eggs. She’d been sneaking back to eat at intervals, then returning to the (impossible) task of waiting for the eggs to hatch.
Honey has stayed around since then, and has built a new nest in amongst the pile of dry kindling in the barn. And for a time, Perky, and our older hens started laying there as well. So there were generally 3 or 4 eggs in that particular nest when I’d check them each day.
Hens don’t usually lay an egg on every day of the week, so when the number of eggs in that nest dropped down to 2 or 3 on most days, I didn’t think too much of it.
Missing eggs + Perky behaving suspiciously in the back paddock = one conclusion.
Last night, Ben shut the gate to the hen enclosure and let the girls out early this morning so that he could see if any of them ran off somewhere.
Believe it or not, there are 19 eggs in this pile!
Sure enough, Perky headed out (the long way) to the back paddock and settled herself down amongst the long grass. Ben found one egg all by itself nearby and left her there to finish laying. When he went back an hour or so later, he found a nest with an additional 19 eggs! Not again! So he brought all the eggs inside and left a fake egg in their place.
At least we know to look there now, and at least Perky hasn’t shown any signs of broodiness. It seems she’s been content to lay an egg on that huge pile, then join her sisters for the rest of the day.
I’m going to check the 20 eggs for freshness, and I may end up discarding a few of them, just to be on the safe side. And if I do… well, that will be the annoying part. The waste of all those beautiful big eggs.
Molly is generally oblivious to the goings-on of the hens.
broody henbrown eggschickenscluckyegg mountainsEggsgardenhensHoneylivestocknesting boxesnestsPerkySouth Headwhere shall I lay my eggs today? | https://heni-irihapeti.com/2015/11/04/egg-mountains/ |
Presented in Partnership with Greene Naftali Gallery
Please join us to celebrate the publication of Experimentations: John Cage in Music, Art, and Architecture with a conversation between the author, Branden W. Joseph and Benjamin Piekut.
Experimentations charts the development of experimental composer John Cage’s aesthetic production from 1940 to 1972. Paying particular attention to Cage’s inter- and cross-disciplinary engagements with the visual arts and architecture during this period, the book sheds new light on some of Cage’s most influential innovations including the use of noise, silence, chance techniques, indeterminacy, electronic technologies, and computerization, as well as investigating lesser-known but important ideas and strategies such as transparency, multiplicity, virtuality, and actualization. Ultimately, it traces the development of Cage’s avant-garde aesthetic and political project as it transformed from an emulation of historical avant-garde precedents such as futurism and the Bauhaus to the development of important precedents for the post-World War II movements of happenings and Fluxus, to its ultimate abandonment in the aftermath of problems encountered in the vast, multimedia composition HPSCHD.
Branden W. Joseph is the Frank Gallipoli Professor of Modern and Contemporary Art at Columbia University, whose work focuses on the relations between the visual arts, music, and film. In addition to Experimentations, he is author of Beyond the Dream Syndicate: Tony Conrad and the Arts after Cage (2008), The Roh and the Cooked: Tony Conrad and Beverly Grant in Europe (2012), Anthony McCall: The Solid Light Films and Related Works (ed. Chris Eamon, 2005), and Random Order: Robert Rauschenberg and the Neo-Avant-Garde (2003), as well as the editor of Kim Gordon, Is It My Body? Selected Texts (2014).
Benjamin Piekut is Associate Professor of Musicology at Cornell University. Student of a wide range of experimental musics in the United States and Europe, he is the author of Experimentalism Otherwise: The New York Avant-Garde and its Limits (2011), editor of Tomorrow is the Question: New Directions in Experimental Music Studies (2014), and co-editor (with George E. Lewis) of The Oxford Handbook of Critical Improvisation Studies (2016). | https://blankforms.org/events/experimentations-john-cage-in-music-art-and-architecture/ |
Hotel Olympic is conveniently located in the popular Diano Marina area. The hotel has everything you need for a comfortable stay. All the necessary facilities, including free Wi-Fi in all rooms, 24-hour front desk, luggage storage, Wi-Fi in public areas, car park, are at hand. Guestrooms are designed to provide an optimal level of comfort with welcoming decor and some offering convenient amenities like air conditioning, heating, balcony/terrace, telephone, satellite/cable TV. The hotel offers various recreational opportunities. For reliable service and professional staff, Hotel Olympic caters to your needs. | https://hotels.intermiles.com/property/hotel-olympic_diano-marina-italy-550647 |
Country’s largest passenger car manufacturer Maruti Suzuki India (MSIL) has reported a net profit of ₹1,166 crore for the fourth quarter ended March 31, lower by around 10 per cent against ₹1,292 crore in the corresponding period last year. The company said the lower income was owing to mark-to-market loss on invested surplus.
However, net sales of the company grew by 33.6 per cent year-on-year (YoY) ₹22,958 crore during the quarter as against ₹17,185 crore in January-March quarter last year. The operating profit for the quarter was at ₹1,250 crore, a growth of 73 per cent over the same period previous year on account of higher sales volume and cost reduction efforts despite steep commodity price increase, the company said in a statement.
The company also said in line with the financial performance of the year and considering uncertain business environment, the Board of Directors recommended a dividend of ₹45 per share (face value of ₹5 per share) for fiscal year (FY) 2020-21.
The Company sold a total of 4,92,235 vehicles during the quarter, higher by 28 per cent compared to the same period previous year. Sales in the domestic market stood at 4,56,707 units, growing by 27 per cent. Exports were at 35,528 units, higher by 44.4 per cent, MSIL said.
“It may be recalled that in the fourth quarter (FY 2019-20) of the previous year there was a significant decline in the sales volume largely owing to Covid-19 lockdown,” it said.
For the full year, net profit for the period stood at ₹4,229 crore, decreasing by 25 per cent compared to that in the previous year on account of lower sales volume, increase in commodity prices, adverse foreign exchange movement, and lower non-operating income partially offset by lower operating expenses, and cost reduction efforts.
During the period, the company registered net sales of ₹66,562 crore, lower by 7.2 per cent compared to that in the previous year.
“The company’s performance for the full year 2020-21 is to be seen in the context of Covid-19 related disruptions,” it said.
MSIL sold a total of 14,57,861 vehicles during the period, lower by 6.7 per cent compared to the FY and lower by 21.7 per cent compared to FY 2018-19. In FY 2020-21, the sales in the domestic market stood at 13,61,722 units, lower by 6.8 per cent and exports were at 96,139 units, lower by 6 per cent compared to the previous year.
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And it has every reason to smile. Covid-19 has triggered a consumer shift towards branded products as ... | https://www.thehindubusinessline.com/companies/maruti-suzuki-q4-net-profit-down-10-to-1166-crore/article34421836.ece |
Are You Lacking A Work/Life Balance?
Many employees, in general, often face problems achieving a work/ life balance.
Many employees, in general, often face problems achieving a work/ life balance.
When new managers start on the job, they usually have a hard task of assembling a team. Whether this requires utilising existing employees, or adding new members, this process is not always as smooth as one would like it to be. Existing employees can be resistant to a change in leadership, and not respect or[…..]
Conflict has a negative image associated with it. Most of us try to stay away from conflict, choosing to separate ourselves from the stress that comes with yelling matches and disagreements.
More and more supervisors are beginning to understand that in order to have loyal and productive staff, they need to take the time to develop relationships with the employees.
When it comes to training a multi-generational workforce, a great debate exists regarding the use of discussions in the training classroom.
One of the greatest challenges in the realm of training is the generational differences found amongst adult learners. While Boomers generally prefer a more traditional classroom setting based on a clear, unidirectional transfer of information from the instructor to the learner, later generations are starting to buck this model and take charge of their learning[…..]
If you are supervising a department or an entire company, as a manager, one of your duties must be to help the business grow.
It is hard for some managers to step back and share control with their staff. Whether the employees have made mistakes in the past, or you simply can’t relinquish control, not having confidence in your team members can lead to negative consequences at the office.
Since the new year started, many organisations have been reviewing their finances from the previous year. When doing so, some may notice that they are not doing as well financially as they would like to. This realisation comes with a hard choice, as one of the easiest options to save money quickly is to downsize.[…..]
Many helpful articles focus on what it takes to be a good leader. However, just as important as it is to strive to develop good qualities, it is just as vital to assess your current style and make sure you have no characteristics of a bad leader. | https://www.mtdtraining.com/blog/2015/03 |
As a result of applied research and our collaboration with industry, we have developed several prototype applications demonstrating the usability and power of technologies, approaches and algorithms we have developed in the Laboratory for Data Technologies. Some of the applications are shortly presented below. If you are interested for more information, please send us an email.
|Supervizor+ is a web application that helps to identify and visualize patterns of relationships among business and physical entities in Slovenia. The application is based on automatic information retrieval from various data sources publicly available on the web, such as AJPES, UJP, ZZS, Supervizor, Wikipedia, Facebook, Google+, LinkedIn, Finance, Dnevnik, Delo, etc. For the implementation of the Supervizor+ we employed Jena framework, a powerful Java framework for building Semantic Web applications, and WebNet component from company Optilab d.o.o. for graph visualization.|
|Occapi is an open intelligent communication platform that has been developed within the OPCOMM competence centre. The purpose of the platform is twofold: (a) to support fusion of large amounts of data, irrespective of their source or structure (e.g. sensors, intelligent devices, metering systems, cameras, web sources, etc.) and (b) to provide users or devices with semantically analyzed and enriched data according to their needs and context. Using the platform, users are able to access semantically enriched data and receive warnings about specific circumstances or events that are recognized by the system. The platform can serve as a foundation for the realization of advanced event and/or data-oriented services and application including well-known paradigms, such as Internet of things (IoT), Smart City, eHealth, eTolling, etc. Based on its modularity, the platform can be extended to support analysis of large amounts of data of any kind and purpose.
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OCCAPI is being developed in conjunction with the Laboratory for Telecommunications. | http://lpt.fri.uni-lj.si/applications |
INTRODUCTION {#sec1-1}
============
Chronic peritoneal dialysis (CPD) is an effective treatment modality in the management of children with end-stage renal failure (ERDS).\[[@ref1][@ref2]\] There are three main techniques for peritoneal dialysis (PD) catheter insertion; open surgical technique, percutanous Seldinger technique, and laparoscopic technique.\[[@ref3][@ref4][@ref5]\] The best technique for inserting PD catheter is still controversial.\[[@ref6]\] Recently, laparoscopic technique has become more popular owing to the shorter hospital stay and more accurate placement of the catheter.\[[@ref7][@ref8]\] Peritonitis and catheter-related infections are still common complications and they are major causes of morbidity.\[[@ref9]\] Peritonitis, pericatheter leakage, exit site infection, failure of fluid drainage due to mechanical dysfunction, and incorrect positioning are the main complications.\[[@ref10]\]
We have developed a single port modified technique of laparoscopic placement of peritoneal dialysis catheter. The purpose of this study was to evaluate the clinical outcomes of this modified laparoscopic technique and open placements of peritoneal catheters.
MATERIALS AND METHODS {#sec1-2}
=====================
Medical records of 49 patients (18 male, 31 female) on PD were reviewed between January 2002 and February 2014. The median age of patients was 15 (range: 3-18 year) in females and 12 (range: 6-17 year) in males. Age, sex, cause of ESRD, catheter insertion method, kt/V rate, peritonitis or other complications, and catheter survival rate were reviewed. The etiology of ESRD was urologic origin in nine patients, nephrologic origin in 20 and etiology could not be determined in 20 patients \[[Table 1](#T1){ref-type="table"}\].
######
Etiologies of end stage renal disease (ESRD).
Kt/V (K; dialyzer clearance of urea, t; dialysis time, V; volume of distribution of urea in the body) is a parameter which is used to quantify the dialysis treatment adequacy.
All surgical procedures were performed under general anesthesia and all patients received a double-cuff curled PD catheter (Kendall, Tyco® peritoneal catheter, Hampshire, UK).
Surgical techniques {#sec2-1}
-------------------
*Open surgical method*; A midline vertical incision was made under the umbilicus and the catheter was inserted into Douglas pouch. Intramuscular tunnel was created up to supra-umbilical area in a semilunar shape. The catheter was passed through the tunnel and exit to the skin. Only the patients, whose omentum is large and mobile enough to come to the incision, underwent omentectomy.
Laparoscopic method {#sec2-2}
-------------------
After diagnostic laparoscopy, the site of the catheter on the right abdominal wall was marked according to the supposed places of the cuffs of the catheter, keeping some distance to the umbilicus. An incision was made on the cranial mark. Afterwards, a subcutaneous tunnel directing caudally was created. The guidebar of the catheter-set was placed through the tunnel into the abdominal cavity. The entrance point of the guidebar into the abdominal cavity at the peritoneum represented the place of caudal cuff of the catheter. Thereafter, a dissector and the catheter itself were placed through the same tunnel. The catheter was placed to the Douglas pouch. An optic forceps made for bronchoscopy replaced the conventional laparoscopy telescope and was inserted through the umbilical port. The catheter was placed its final position. In all cases, the omentum was grasped by the optic forceps. The umbilical trocar is removed, while optic forceps grasping the omentum was kept in situ. The optic forceps with omentum were taken out through the trocar-site and the omentum was excised ex-corpo.
Peritonitis was defined as clinical features (abdominal pain, fever, and cloudy dialysate) and leukocytosis in dialysis solution (white blood cell count \>1000 µl with \>50% neutrophils).\[[@ref11]\]
In the case of peritonitis, the initial management is always conservative. Withdrawal of catheter is only indicated in patients with failed conservative treatment of systemic antibiotics or recurrent peritonitis attacks.
The application of dialysis before 2 weeks subsequent to catheter placement was defined as "early usage" and after 2 weeks as "late usage."
Statistical analysis {#sec2-3}
--------------------
Descriptic statistics were performed. For comparison of groups, ANOVA or Kruskal-Wallis variance analysis, Chi-square test or Fishers' exact test, and Mann-Whitney U test were used after normality analysis.
RESULTS {#sec1-3}
=======
Thirty-three of the catheters were placed by open method (21 female and 12 male) whereas 16 were placed by laparoscopy (10 female and 6 male). The mean age of the patients was 11.95 ± 0.61 (12.12 ± 0.75 in open group; 11.62 ± 1.05 in the laparoscopic group). There was no difference between open and laparoscopic methods, as well as between males and females.
Total of 29 patients had peritonitis (59.1%) (16 females and 13 males) and only four of them developed after laparoscopic placement (25% in laparoscopic 75% in open group). The frequency of the peritonitis was significantly less in patients with laparoscopic access when compared to those with open method (*P* = 0.001). Patients with peritonitis were younger (10.95 ± 0.8) than those without peritonitis (13.4±0.85) (*P* = 0.049) \[[Table 2](#T2){ref-type="table"}\]. There was no significant difference in frequency of peritonitis between patients with underlying urologic or nephrological problems (*P* = 0.167) \[[Table 3](#T3){ref-type="table"}\]. Initially, all patients with peritonitis were treated conservatively, but in nine patients with peritonitis, the conservative management failed and we had to remove these patients' catheters.
######
Mean age and length of PD measurement according to PD technique, presence of peritonitis
######
Number of peritonitis and other complications according to PD technique and etiology of ESRD
In nine patients, the catheter was used early after placement. Three of them had peritonitis. Dialysate leakage was not observed in patients with early usage. Forty patients were in the "late use" group and 23 of them developed peritonitis. There was no significant difference between early and late usage of PD according to development of peritonitis (*P* = 0.455).
Other complications were in total of 12 patients (24.5 %) including catheter leakage (*n* = 5), catheter obstruction leading to inadequate inflow and outflow (*n* = 4), exit-site or tunnel infection (*n* = 3). No significant difference was found between the open (nine complications) and laparoscopic (three complications) groups in terms of complication development (*P* = 0.541). Complication frequency was not different between patients with urological and nephrological problems either (*P* = 0.406) \[[Table 3](#T3){ref-type="table"}\]. In the laparoscopic group, two patients had catheter obstruction and one patient had a leakage. Patients with leakage were treated conservatively (Decreasing filling volume, switch to automated peritoneal dialysis temporarily). From the patients with catheter obstruction, three catheters were revised and the remaining one recovered without surgical intervention. One patient had catheter migration; his catheter was removed. Patients with exit-site or tunnel infection were treated conservatively.
Five patients were converted to hemodialysis due to recurrent peritonitis. None of them belonged to the laparoscopic group. Renal transplantation was performed in 12 patients. Inguinal hernia was developed in four male patients subsequently and one of them was bilateral.
Catheter survival rate for the first and second years was 95% in both groups and catheter survival rate for the 5 years were 87.5% in the open group. Since inadequate number of patients in the laparoscopic group had surveyed longer than 5 years, we are not able to measure catheter survival rate for 5 years in those patients.
The mean kt/V which indicates the effectiveness of peritoneal dialysis was 2.26 ± 0.08 in all patients. kt/V values were not different between laparoscopic and open approach groups (2.20 ± 0.17 vs 2.28 ± 0.1) (*P* = 0.416).
Three patients died due to septic complications and peritonitis. All of them were in the open group. There was no significant difference of survival between open and laparoscopic groups.
Microbiologic analysis of the blood and/or peritoneal fluid of those with peritonitis is shown in [Table 4](#T4){ref-type="table"}.
######
Number of positive cultures in patients with peritonitis
DISCUSSION {#sec1-4}
==========
PD is an effective and alternative method for renal replacement therapy in children with ESRD to enhance patient\'s survival and quality of life. The major advantage of the PD is its easy use as home setting by caregivers.\[[@ref12][@ref13]\] There are three main methods available for PD placement including open, laparoscopic approaches, and Seldinger method. In all types of placements, complications may occur such as catheter obstruction and leakage, tunnel infection, or peritonitis. These complications may decline the quality of life of the patients and duration of catheter.\[[@ref14][@ref15][@ref16]\]
Laparoscopic catheter placement for PD provides direct vision, minimal invasion, and accurate positioning of catheter into pelvis\[[@ref17][@ref18][@ref19]\] as well as a more proper omentectomy. Many investigators suggested that laparoscopic catheter placement is superior to open method in terms of the complication rate (especially peritonitis) and catheter survival.\[[@ref8][@ref20]\] In this study, we did not detect such advantages except peritonitis which was very low in laparoscopic group (25 in open group vs 4 in laparoscopic group). The cause of dialysis associated peritonitis in children is not clear. Contamination, training, prophylactic antibiotics, patient age were primary factors that affect the incidence of peritonitis. The incidence of peritonitis was less in laparoscopic catheter insertion group. Accurate placement with direct vision provides more appropriate positioning of the catheter. Omentectomy was performed in all of our patients with laparoscopic access by direct vision. Laparoscopic insertion favors in terms of omentectomy because of better vision of omentum. We think omentectomy decreases complication rate particularly related to obstruction and peritonitis,\[[@ref15][@ref21]\] though survival rates were similar in both groups in the current study. The reason might be shorter duration of follow up for patients with laparoscopic catheter placement.
Many studies suggested that laparoscopic catheter placement has no obvious advantages over open placement according to complication rate, cost-effectiveness, and length of procedure.\[[@ref17][@ref22][@ref23]\] However, we suggest laparoscopic placement has lower peritonitis rate and less incidence of catheter migration due to the ability of direct vision of peritoneal cavity. Similarly, some authors\[[@ref24][@ref25]\] claim that placement modality catheter survival had no superiority on catheter survival, but some others\[[@ref2][@ref26]\] suggest that laparoscopic placement had higher catheter survival.
Many technical modifications have been described for the laparoscopic placement.\[[@ref27]\] Our modified method provides an easier placement. The usage of optic forceps for the laparoscopic insertion of peritoneal dialysis catheter facilitates the dissection. It reduces the need for additional trocar and provides appropriate position of the catheter. The omentectomy is much simpler than other laparoscopic placement methods.
In conclusion, catheter survival rate was excellent both in open and single port laparoscopic surgery groups. None of the method is complication free and rates of complications are similar in both groups except the peritonitis which is significantly lower in the laparoscopic group. The single port laparoscopic method for the CPD seems to be safe, effective, and cosmetically superior. Laparoscopic method for CPD may lead to less peritonitis episodes.
**Source of Support:** Nil
**Conflicts of Interest:** None declared.
| |
FIELD OF THE INVENTION
BACKGROUND
SUMMARY OF INVENTION
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS
Definitions
EXAMPLES
Example 1
Materials and Methods
Tamoxifen Injection and Tissue Collection
Cardiomyocyte Isolation
Histological and Immunofluorescence Microscopy
Western Analysis for LMNA, SUN1, Ha-Tag and GFP.
Active Force Measurement of Cardiac Papillary Muscle.
AAV9-N-Sun1 and AAV9-GFP Virus
Generation of Mutant Mice Using CRISPR/Cas9
DNA Extraction for Genotyping of CRISPR/Cas9 Mice
Genotyping of CRISPR/Cas9 Mice
Derivation of Myoblasts, Fibroblasts and Cell Culture for CRISPR/Cas9 Study
Immunoblotting for CRISPR/Cas9 Study
Immunofluorescence for CRISPR/Cas9 Study
Mouse Genetics
Human Guide RNA Sequences
Statistical Analysis
Example 2
Cardiomyocyte Specific Loss of Lmna Results in the Rapid Onset of Heart Failure
Example 3
Deletion of Sun1 Amellorates Cardiac Pathology Induced by Lmna Loss
Example 4
Loss of SUN1 Extends Longevity of Lmna Missense Mutants
Example 5
Example 6
Disruption of the LINC Complex in Mice Using CRISPR/Cas9
Characterization of Mutant Mice
Disruption of Nesprin-1 KASH Domain Ameliorates Lmna Pathologies
Example 7
DISCUSSION
BIBLIOGRAPHY
The present invention relates to use of expression vectors and other compounds in methods to disrupt the Linker of Nucleoskeleton and Cytoskeleton (LINC) complex, uncoupling the nucleus from its linkage to the cytoskeleton, resulting in amelioration of diseases caused by one or more Lmna mutations, so-called laminopathies. More particularly, the invention relates to the expression of dominant negative or mutated SUN domain protein and/or dominant negative or mutated KASH domain protein to disrupt the LINC complex in, for example, cardiomyocytes for suppressing disease progression in dilated cardiomyopathy (DCM).
Lancet
Nat Rev Cardiol
N Engl J Med
Genome Med
Genome Med
Dilated Cardiomyopathy (DCM) is the most common disease affecting heart muscle, accounting for approximately 60% of all cardiomyopathies. It is characterized by reduced systolic (contractile) function due to enlargement and thinning of the left ventricular wall or in some cases both ventricles. DCM is associated with sudden heart failure and cardiac death, resulting in high rates of hospital admission, the need for heart transplantation and consequently a high cost burden [J. L. Jefferies and J. A. Towbin, 375:752-762 (2010); R. E. Hershberger, et al., 10: 531-547 (2013)]. The causes of DCM are varied, but include a variety of extrinsic factors, (viral, autoimmune infiltration, alcohol, and drugs). However 30-40% of all cases have a monogenic basis, with mutations in some 40 genes being linked to DCM. The most frequently mutated gene in DCM is TTN, that encodes the giant sarcomeric protein titin, with truncating variants in TTN accounting for almost 15-25% of all congenital forms of DCM [D. S. Herman et al., 366: 619-628 (2012); U. Tayal, S. et al., 9: 20 (2017)]. The second most frequently mutated gene is Lamin A (LMNA) accounts for as many as 6-8% of congenital DCM patients [U. Tayal, S. et al., 9: 20 (2017)].
Hum Mol Genet
Gene Reviews
R
Heart
LMNA Induced DCM Is characterized by cardiac conduction disease manifested by electrophysiological (ECG) abnormalities, including atrioventricular block, ventricular arrhythmias and fibrillation. The risk of sudden cardiac death is greater in patients with LMNA-cardiomyopathy than patients with other forms of DCM [J. H. Van Berlo et al., 14: 2839-2849 (2005)]. Some 450 different mutations have been Identified in the LMNA gene, most being missense, resulting in the majority of DCM cases being inherited as autosomal dominants, with this diversity of the mutations complicating genetic approaches to treating LMNA induced DCM. To a limited extent LMNA induced DCM can be treated by fitting a pacemaker. Ultimately, however, effective treatment at present is accomplished by heart transplantation (R. E. Hershberger and A. Morales, in (()), M. P. Adam et al., Eds. (Seattle (Wash.), 1993); G. Captur et al., 104: 488-479 (2018)].
J Cell Biol
Hum Mol Genet
J Clin Invest
Differentiation; research in biological diversity
J Clin Invest
Hum Mol Genet
Mouse lines carrying Lmna mutations usually die within a few weeks after birth [T. Sullivan et al., 147: 913-920 (1999); A. T. Bertrand et al., 21: 1037-1048 (2012); V. Nikolova et al., 113: 357-369 (2004); A. S. Wang, et al., , (2015)]. The cause of early death in mice lacking Lmna is uncertain due to multiple tissues being affected. Cardiac myopathy is thought to be a major contributing cause, as Lmna mutant mice develop DCM with conduction abnormalities and focal myocyte degeneration [V. Nikolova et al., 113: 357-369 (2004); L. C. Mounkes, et al., 14: 2167-2180 (2005)], although effects on other, as yet undefined, skeletal muscles may contribute to early postnatal death.
Nat Rev Mod Cell Biol
J Cell Biol
Cell
Cell
Cold Sprng Harbor perspectives in biology
Exp Cell Res
J Cell Biol
Cell
Cell
Hum Mol Genet
J Biol Chem
Cell
Cell
Cell
Cell
−/−
−/−
−/−
−/−
−/−
−/−
−/−
−/−
−/−
−/−
−/−
The lamins are nuclear intermediate filament proteins and are the principal constituents of the nuclear lamina, the proteinaceous matrix underlying the inner nuclear membrane (INM). The lamina consists of the A-type lamins, consisting of 2 predominant forms, lamins A and lamin C, derived by alternate splicing of LMNA, whereas the two B type lamins (LMNB 1 and 2) are each encoded by two genes: LMNB1 and LMNB2 [B. Burke and C. L. Stewart, 14: 13-24 (2013)]. The lamina provides structural and mechanical integrity to the nucleus, maintains nuclear shape and position within the cell, as well as being determinants of chromatin organization [T. Sullivan et al., 147: 913-920 (1999); I. Solovei et al., 152: 584-598 (2013)]. The lamins interact with numerous INM proteins, inducing Emerin, the Lamina-Associated Polypeptides (LAPs) and the SUN domain proteins [B. van Steensel and A. S. Belmont, 169: 780-791 (2017)], many of which when either mutated or present as a variant are linked to heart disease [H. J. Worman, et al., 2: a000760 (2010); C. L. Stewart, et al., 313:2144-2156 (2007)]. Together these proteins comprise an integrated protein network, centered on the lamina, where loss or mutation of the lamins can result in either the mislocalization or a change in expression levels of many lamina associated proteins, (emerin, SUN1, LBR and Lap2α) [T. Sullivan et al., 147: 913-920 (1999); I. Solovel et al., 152: 584-598 (2013); C. Y. Chen et al., 149: 565-577 (2012); T. V. Cohen et al., 22: 2852-2869 (2013); F. Haque et al., 285: 3487-3498 (2010)]. Among these proteins, where expression is affected by the loss of Lmna or mutation, are SUN1 and Lap2, both of whose levels are Increased. In the case of SUN1 the Increased level is due to reduced turnover, rather than increased expression, resulting in high levels accumulating in the Golgi which appeared to be cytotoxic at least in the Lmna and LmnaΔ9 mouse disease models [C. Y. Chen et al., 149: 565-577 (2012); C. Stewart and B. Burke, WO/2013/158046]. However, when SUN1 levels are genetically ablated in mice with Lmna mutations, this increases the longevity 3-fold and ameliorates much of the pathology [C. Y. Chen et al., 149: 585-577 (2012); C. Stewart and B. Burke, WO/2013/158048]. The median survival of wild-type or Sun1 is >210 days in a 7 month follows up; Lmna mice had median survival of 41 days; Lmna Sun1 mice had a median survival of 54 days; Lmna Sun1 mice had a median survival of 104 days (p<0.01 comparing Lmna and Lmna Sun1). Likewise, whereas all LmnaΔ9 mice expired by 30 days after birth, their LmnaΔ9Sun1 littermates thrived past this date, and most achieved life spans more than twice this duration [C. Y. Chen et al., 149: 565-577 (2012)]. At the cellular level, human fibroblasts harbouring a LMNA mutation resulting in Hutchison-Gilford Progeria Syndrome also exhibited increased Sun1 levels. Depleting Sun1 in these cells alleviated nuclear morphology defects, again suggesting that excess Sun1 resulting from LMNA mutation is cytotoxic [C. Y. Chen et al., 149: 565-577 (2012); C. Stewart and B. Burke, WO/2013/158046]
J Cell Biol
Curr Opin Cell Biol
Current topics in developmental biology
Scientific reports
Proc Natl Acad Sci USA
The SUN (Sad1p, UNC-84) domain proteins share a conserved C-terminal SUN domain and localize to the INM [C. J. Malone, et al., Development 126:3171-3181 (1999)]. In mammals, SUN1 and SUN2 are the 2 principal SUN proteins that are widely expressed in virtually all tissues. In the perinuclear space, between the INM and outer nuclear membrane (ONM), the C-termini of SUN1 and/or 2 bind to the C-termini (KASH domains) of the different Nesprins/SYNE/KASH proteins that traverse the ONM. Together these 2 families of proteins comprise the LINC complexes that physically couple the interphase nuclei to the cytoskeleton [M. Crisp et al., 172: 41-53 (2006); E. C. Tapley and D. A. Starr, 25: 57-62 (2013)]. The N-termini of the SUN domain proteins protrude into the nudeoplasm and with SUN1, this region interacts with pre-laminA and nuclear pore complexes. Whether the N-terminus of SUN2 interacts with any nucleoplasmic/NE protein is unclear. In contrast, the bulk of the Nesprins/KASH domain proteins extend into the cytoplasm adjacent to the ONM. There, depending on the particular Nesprin/KASH protein, they interact directly or indirectly with all 3 cytoskeletal protein networks (microtubules, actin microfilaments and Intermediate filaments) [H. F. Horn, 109: 287-321 (2014)]. Together, the SUN and KASH/Nesprin proteins of the LINC complex establish a direct physical connection between the cytoplasmic cytoskeletal networks (and their connections e.g. cell adhesion complexes at the cell membrane) and the interphase nuclear interior or nucleoplasm. The LINC complex Is thought to mediate force transmission between the nucleus and cytoskeleton and consequently regulate changes in gene expression/chromatin organization in response to mechanical/physical stimuli [S. G. Alam et al., 6: 38003 (2016)]. Although loss of either SUN1 or SUN2 alone has no overt effect on postnatal growth and viability, SUN1 null mice are infertile and deaf. Simultaneous loss of Sun1 and Sun2 results in perinatal lethality, indicating a degree of redundancy during embryogenesis [K. Lei et al., 106: 10207-10212 (2009)].
There is a need to develop alternative methods to ameliorate the negative effects over-accumulation of Sun1 has on cells carrying Lmna mutations. The present disclosure aims at providing such a method.
Surprisingly, the inventors have found that disruption of the LINC complex rather than removal of accumulated Sun1 protein can ameliorate diseases caused by one or more Lmna mutations. One way of achieving the disruption is via an expression construct/vector comprising an operably linked transgene, the expression of which generates dominant negative SUN domain protein or mutated endogenous SUN domain protein and/or dominant negative KASH domain protein or mutated endogenous KASH domain protein. The exogenous dominant negative SUN domain and KASH domain proteins act as LINC complex binding competitors, thereby uncoupling the nucleus from its linkage to the cytoskeleton. The mutated SUN domain and KASH domain proteins are endogenous Sun and Nesprin proteins that have been mutated in the SUN or KASH domain, respectively, and cannot form a UNC complex because they cannot bind to their cognate LINC complex partner. These strategies may be used to disrupt the LINC complex to treat, for example, laminopathies. The result was achieved without actively reducing the endogenous SUN1 protein levels. Results shown herein support these claims.
According to a first aspect of the invention, there is provided an isolated nucleic acid molecule, wherein the nucleic acid molecule comprises an expression vector and a transgene, whereby the transgene is operably linked to the expression vector, wherein expression of the transgene in a transfected cell results in disruption of a LINC complex in the transfected cell.
In some embodiments, the expression vector is a cardiac- or cardiomyocyte-specific expression vector.
In some embodiments, the expression vector comprises a cardiac- or cardiomyocyte-specific promoter selected from the group comprising a cardiac troponin T promoter (cTnT), a α-myosin heavy chain (α-MHC) promoter and a myosin light chain (MLC2v) promoter. Preferably the promoter is cardiac troponin T promoter (cTnT).
In some embodiments, the cardiomyocyte-specific promoter is chicken cardiac troponin T promoter (cTnT).
In some embodiments, the expression vector has cardiac tropism/is cardiotropic.
In some embodiments, the expression vector is a virus expression vector.
In some embodiments, the virus expression vector Is selected from the group comprising Lentivirus, Adenovirus and Adeno-associated virus (AAV). Preferably the virus expression vector is adeno-associated virus (AAV).
In some embodiments, the AAV vector is selected from the group consisting of AAV9 (serotype 9), AAV1 (serotype 1), AAV6 (serotype 6), AAV8 (serotype 8), AAV218 and AAV9.45.
In some embodiments, the AAV vector is AAV9 (serotype 9).
In some embodiments, the transgene comprises nucleic acid sequences for expressing a lumenal domain of a SUN domain-containing protein, an N-terminal signal sequence, a signal peptidase cleavage site, and a C-terminal targeting peptide sequence.
In some embodiments the lumenal domain of the SUN domain-containing protein comprises a coiled coil domain and a SUN domain.
In a preferred embodiment the coiled coil domain is upstream of the SUN domain.
In some embodiments, the transgene further comprises nucleic acid sequences for expressing an N-terminal signal sequence, a signal peptidase cleavage site, and a C-terminal targeting peptide sequence.
In some embodiments, the transgene comprises nucleic acid sequences for expressing an N-terminal signal sequence, a signal peptidase cleavage site, and a C-terminal targeting peptide sequence, and either the luminal domain of the SUN domain-containing protein or the SUN domain.
Preferably, the SUN domain protein is SUN1 or SUN2.
FIG. 7
In some embodiments the luminal domain of Sun1 comprises amino acids 458-913 of full-length mouse Sun1 (Uniprot Q9D666) or Its human equivalent comprising the coiled coil domain and the SUN domain and lacking the transmembrane domain. A schematic of the structure of a dominant negative form of Sun is shown in .
Cell
For SUN domain constructs it is expected that the SUN domain alone (crystal structure solved by the Kutay and Schwartz labs [Sosa et al., 149(5):1035-47 (2012)], Instead of the entire luminal domain (coiled coil domain and SUN domain) is sufficient to disrupt the SUN-KASH interaction as it is capable of binding to the KASH domain. The human Sun1 SUN domain nucleic acid sequence is set forth in SEQ ID NO: 80. However, the presence of the signal sequence and the KDEL sequence are important for targeting the construct to the perinuclear space.
In some embodiments, the N-terminal signal sequence is derived from a secretory protein or a Type I transmembrane protein.
Preferably, the secretory protein or Type I transmembrane protein is selected from the group consisting of human serum albumin, proinsulin, transferrin receptor, EGF receptor, pre-pro-opiomelanocortin, pancreatic digestive enzymes (for example, proteases, amylases and lipases), endoplasmic reticulum luminal proteins, for example protein disulphide isomerases, GRP94 and combinations thereof. More preferably, the N-terminal signal sequence is derived from human serum albumin.
In some embodiments, the N-terminal signal sequence is not preceded at its N-terminus by any other tags.
In some embodiments, the signal peptidase cleavage site is a signal peptidase cleavage site derived from or is one of the group consisting of human serum albumin, proinsulin, transferrin receptor, EGF receptor, pre-pro-opiomelanocortin, pancreatic digestive enzymes (for example, proteases, amylases and lipases), endoplasmic reticulum lumenal proteins, such as protein disulphide isomerases, GRP94 and combinations thereof. Preferably, the signal peptidase cleavage site is a signal peptidase cleavage site derived from human serum albumin.
In some embodiments, the C-terminal targeting peptide sequence prevents secretion of a peptide expressed from the transgene according to any aspect of the invention.
FIGS. 11 and 12
In some embodiments, the C-terminal targeting peptide sequence is a KDEL tetrapeptide Golgi retrieval sequence. Examples of such structures are shown in .
In some embodiments the transgene comprises a humanized Sun1DN nucleic acid sequence or a humanized Sun2DN nucleic acid sequence. In a preferred embodiment, the transgene comprises a signal sequence, a humanized Sun1DN nucleic acid sequence and a KDEL sequence as set forth in SEQ ID NO: 4; or the transgene comprises a signal sequence, a humanized Sun2DN nucleic acid sequence and a KDEL sequence as set forth in SEQ ID NO: 5.
In some embodiments, the transgene further comprises an epitope tag. Preferably the epitope tag is N-terminal, or located anywhere in the nucleic acid molecule except downstream of (after) the C-terminal targeting peptide sequence [for example KDEL], or located anywhere in the nucleic acid molecule except upstream of (before) the N-terminal signal sequence.
In some embodiments, the epitope tag is selected from the group consisting of cellulose binding domain (CBD), chloramphenicol acetyl transferase (CAT), dihydrofolate reductase (DHFR), one or more FLAG tags, glutathione S-transferase (GST), green fluorescent protein (GFP), haemagglutinin A (HA), histidine (His), Herpes simplex virus (HSV), luciferase, maltose-binding protein (MBP), c-Myc, Protein A, Protein G, streptavidin, T7, thioredoxin, V5, vesicular stomatitis virus glycoprotein (VSV-G), and combinations thereof. Preferably, the epitope tag is haemagglutinin A (HA).
In some embodiments, the nucleic acid molecule of the invention comprises an adeno-associated virus vector (AAV) comprising a chicken cardiac troponin T promoter (cTnT), a transgene according to any aspect of the invention comprising the luminal domain of the SUN domain-containing protein derived from SUN, an N-terminal signal sequence and a signal peptidase cleavage site which are each derived from human serum albumin, a C-terminal targeting peptide sequence which is a KDEL sequence, and wherein the transgene optionally further comprises haemagglutinin (HA) as an N-terminal epitope tag.
FIG. 10
According to an embodiment an example of such a vector is shown in and comprises the nucleic acid sequence set forth in SEQ ID NO: 3.
In some embodiments, the nucleic acid molecule of the invention comprises an adeno-associated virus vector (AAV) comprising a chicken cardiac troponin T promoter (cTnT), a transgene according to any aspect of the invention comprising the luminal domain of the SUN domain-containing protein derived from SUN2, an N-terminal signal sequence and a signal peptidase cleavage site which are each derived from human serum albumin, a C-terminal targeting peptide sequence which is a KDEL sequence, and wherein the transgene optionally further comprises hemagglutinin (HA) as the N-terminal epitope tag.
FIG. 10
According to an embodiment an example nucleic acid molecule would comprise the vector structure shown in and the transgene nucleic acid sequence set forth in SEQ ID NO: 5.
Rather than expressing components of a lumenal domain of a SUN domain-containing protein, a KASH domain may be expressed to disrupt a LINC complex by competing with endogenous Nesprins (which comprise a KASH domain) for binding to SUN1 and SUN2 domains.
Accordingly, in some embodiments of the nucleic acid molecule of the Invention, the transgene comprises nucleic acid sequences for expressing a KASH domain, and an N-terminal stabiliser polypeptide sequence.
Preferably, the KASH domain comprises a transmembrane domain and a SUN-interacting peptide.
Preferably the transgene comprises nucleic acid sequences for expressing a KASH domain that traverses the outer nuclear membrane, a SUN-interacting KASH peptide that extends into the perinuclear space at the C-terminus, and an N-terminal stabiliser polypeptide sequence in the cytoplasm.
It would be understood that KASH domain constructs with extensions after the last C-terminal amino acid of the naturally occurring KASH domain are not expected to work. i.e. C-terminal tags, or even an additional carboxy-terminal single amino acid, will disrupt KASH interaction with SUN. In addition, a signal sequence on the N-terminus of SUN domain constructs cannot be preceded by any tags.
In some embodiments, the KASH domain is selected from the group consisting of KASH1 (derived from Nesprin-1 (SYNE1 gene)), KASH2 (derived from Nesprin-2 (SYNE2 gene)), KASH3 (derived from Nesprin-3 (SYNE3 gene)), KASH4 (derived from Nesprin-4 (SYNE4 gene)) and KASH5 (derived from KASH5/CCDC155 (KASH5 gene)).
FIG. 14
In preferred embodiments the KASH 1 domain comprises the human amino acid sequence set forth in SEQ ID NO: 7; the KASH 2 domain comprises the human amino acid sequence set forth in SEQ ID NO: 9; the KASH 3 domain comprises the human amino acid sequence set forth in SEQ ID NO: 11; the KASH 4 domain comprises the human amino acid sequence set forth in SEQ ID NO: 13; and the KASH 5 domain comprises the human amino acid sequence set forth in SEQ ID NO: 15. An alignment of the five KASH amino acid sequences is shown in .
In some embodiments the KASH domain nucleic acid sequence has at least 80%, at least 85%, at least 90%, at least 95% sequence identity or 100% sequence identity to the nucleic acid sequence of the KASH1 domain set forth in SEQ ID NO: 6; the nucleic acid sequence of the KASH2 domain set forth in SEQ ID NO: 8; the nucleic acid sequence of the KASH3 domain set forth in SEQ ID NO: 10; the nucleic acid sequence of the KASH4 domain set forth in SEQ ID NO: 12; or the nucleic acid sequence of the KASH5 domain set forth in SEQ ID NO: 14.
More preferably, for the purpose of clinical use, the KASH domain is the human KASH1 domain of SYNE1 having at least 80%, at least 85%, at least 90%, at least 95% sequence identity or 100% sequence identity to the nucleic acid sequence of the human KASH1 domain set forth in SEQ ID NO: 6.
It would be understood that due to the redundancy in the genetic code, a nucleic acid sequence may have less than 100% identity and still encode the same amino acid sequence.
In some embodiments, the KASH domain does not comprise any extensions after the last C-terminal amino acid compared to a naturally occurring KASH domain.
In some embodiments, the N-terminal stabiliser polypeptide sequence Is selected from the group consisting of green fluorescent protein (GFP), cellulose binding domain (CBD), chloramphenicol acetyl transferase (CAT), dihydrofolate reductase (DHFR), glutathione S-transferase (GST), luciferase, maltose-binding protein (MBP), Protein A, Protein G, streptavidin, thioredoxin, DHFR, including multiples and combinations thereof.
In some embodiments, the N-terminal stabilizer polypeptide sequence forms a discretely folded domain.
In some embodiments, the vector is the adeno-associated virus vector (AAV) comprising a cardiac troponin T promoter (cTnT), and the transgene comprises nucleic acid sequences for expressing a KASH domain, and an N-terminal stabiliser polypeptide sequence, wherein the KASH domain is selected from the group comprising KASH1, KASH2, KASH3, KASH4 and KASH5.
In some embodiments, the N-terminal stabiliser polypeptide sequence is green fluorescent protein (GFP).
Rather than expressing components of a lumenal domain of a SUN domain-containing protein or a KASH domain to disrupt a LINC complex by competing for binding with endogenous Nesprins (which comprise a KASH domain) or Sun1 and Sun2 (which comprise a SUN domain), another approach for disrupting the LINC complex is to modify the endogenous SUN domain or KASH domain so that it fails to bind to, or has reduced binding capacity for, its cognate LINC complex binding partner.
As both the SUN domain and the KASH domain are located at the C-termini of their respective proteins, one way of producing a modified SUN or KASH domain is to use a CRISPR/Cas system to modify the genes encoding SUN or KASH domain proteins to generate a premature stop codon at the 3′ end of the respective protein sequences following CRISPR-induced non-homologous end joining. This would result in a truncated protein with its C-terminal SUN or KASH domain mutated. The truncated protein would be expressed and membrane-localized, but unable to interact with its cognate LINC complex partners.
Accordingly, in some embodiments of the nucleic acid molecule of the invention, the transgene comprises nucleic acid sequences for expressing a CRISPR-Cas or other synthetic nuclease system to modify nucleic acid that encodes the SUN domain or KASH domain of endogenous Sun or Nesprin protein, respectively.
Data shown herein (Example 6) suggests that modification of the SUN2 domain or KASH2 domain does not ameliorate Lmna pathology.
In some embodiments the CRISPR-Cas modifies the endogenous SUN domain or KASH domain of Sun1 or Nesprin-1 protein, respectively, to disrupt a LINC complex. The respective nucleic acids are Sun1 and Syne1.
In some embodiments, the transgene comprises nucleic acid sequences for expressing a CRISPR-Cas with a gRNA nucleic acid sequence comprising 5′-GCACAATAGCCTCGGATGTCG-3′ (SEQ ID NO: 66) to modify the SUN domain of mouse Sun1.
In some embodiments, the transgene comprises nucleic acid sequences for expressing a CRISPR-Cas with a gRNA nucleic acid sequence targeting the human SUN1 domain set forth in SEQ ID NO: 80. Preferably, the gRNA nucleic acid sequence targets the end of axon 20 comprising a nucleic acid sequence set forth in SEQ ID NO: 81. More preferably, the gRNA nucleic acid sequence targets a SUN1 nucleic acid sequence selected from the group comprising SEQ ID NO: 55; SEQ ID NO: 56; SEQ ID NO: 57; SEQ ID NO: 58; SEQ ID NO: 59; SEQ ID NO: 60; SEQ ID NO: 61; SEQ ID NO: 62; SEQ ID NO: 63; SEQ ID NO: 64 and SEQ ID NO: 65 set forth in Table 3.
In some embodiments, the transgene comprises nucleic acid sequences for expressing a CRISPR-Cas with a gRNA nucleic acid sequence comprising 5′-CCGTTGGTATATCTGAGCAT-3′ (SEQ ID NO: 34) to modify the KASH domain of mouse Syne-1.
In some embodiments, the transgene comprises nucleic acid sequences for expressing a CRISPR-Cas with a gRNA nucleic acid sequence targeting the human KASH domain set forth in SEQ ID NO: 6. Preferably, the gRNA nucleic acid sequence comprises a nucleic sequence selected from the group comprising SEQ ID NO: 44; SEQ ID NO: 45; SEQ ID NO: 46; SEQ ID NO: 47; SEQ ID NO: 48; SEQ ID NO: 49; SEQ ID NO: 50; SEQ ID NO: 51; SEQ ID NO: 52; SEQ ID NO: 53 and SEQ ID NO: 54 set forth in Table 3.
In some embodiments, the transgene comprises nucleic acid sequences for expressing a CRISPR-Cas9 or variant thereof.
In preferred embodiments, the transgene is a dominant negative construct.
In some embodiments, the transgene is a humanised transgene.
−/−
−/−
−/−
In some embodiments, expression of the transgene results in the disruption of the protein-protein Interaction between SUN and KASH domains of the LINC complex. Preferably, the disruption of the protein-protein interaction between SUN and KASH of the LINC complex occurs between the protein interactions selected from the group consisting of Sun1 Nesprin-1, Sun2+Nesprin-1, Sun1 Nesprin-2, Sun1 Nesprin-3, Sun2+Nesprin-2, and Sun2+Nesprin-3. More preferably, the disruption of the protein-protein interaction between SUN and KASH of the LINC complex occurs between the protein interactions of Sun1 and Nesprin-1.
In some embodiments, the AAV vector is formulated for delivery into the myocardium of a subject.
According to a second aspect of the invention there is provided a nucleic acid molecule of any embodiment of the invention for use in treating a disease caused by one or more Lmna mutations in a subject.
In some embodiments of the second aspect, the disease is selected from the group consisting of restrictive dermopathy, familial partial lipodystrophy (for example, Dunnigan type), mandibuloacral dysplasia with type A lipodystrophy, metabolic syndrome, Charcot-Marie-Tooth disease type 2, Charcot-Marie-Tooth disease type 2B1 and diseases presented in normal font in Table 1.
TABLE 1
Database
cDNA Variant
Protein Variant
Disease
Omim
ID
cDNA Variant
Types
Protein Variant
Types
Domain
Abbreviation
Disease Name
Omim ID
Symbol
17811
c.1771T > A
Substitution
p.Cys591Ser
Substitution
Tail
—
Acrogeria, Gottron Type
201200
#
18255
c.418_438dupCTGCTG
Duplication
p.Leu140_Ala146dup
Duplication
1B
—
Arrhythmogenic
—
—
AACTCCAAGGAGGCC
cardiomyopathy
17439
c.1039G > A
Substitution
p.Glu347Lys
Substitution
2B
ARVD7
Arrhythmogenic right
609160
%
ventricular cardiomyopathy
18492
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
ARVD7
Arrhythmogenic right
609160
%
ventricular cardiomyopathy
9157
c.1718C > T
Substitution
p.Ser573Leu
Substitution
Tail
—
Arthropathy syndrome,
—
—
autosomal recessive
18299
c.1494G > A
Substitution
p.Trp498X
Substitution
Tail
AF
Atrial fibrillation
—
—
18301
c.175C > G
Substitution
p.Leu59Val
Substitution
1A
APS
Atypical progeroid syndrome
—
—
8885
c.169G > C
Substitution
p.Ala57Pro
Substitution
1A
WRN
Atypical Werner syndrome
277700
#
8886
c.398G > T
Substitution
p.Arg133Leu
Substitution
1B
WRN
Atypical Werner syndrome
277700
#
11462
c.398G > T
Substitution
p.Arg133Leu
Substitution
1B
WRN
Atypical Werner syndrome
277700
#
9232
c.398G > T
Substitution
p.Arg133Leu
Substitution
1B
WRN
Atypical Werner syndrome
277700
#
11509
c.398G > T
Substitution
p.Arg133Leu
Substitution
1B
WRN
Atypical Werner syndrome
277700
#
11670
c.398G > T
Substitution
p.Arg133Leu
Substitution
1B
WRN
Atypical Werner syndrome
277700
#
8888
c.419T > G
Substitution
p.Leu140Arg
Substitution
1B
WRN
Atypical Werner syndrome
277700
#
13457
c.506delT
Deletion
p.Val169GlyfsX7
Frame shift
1B
WRN
Atypical Werner syndrome
277700
#
13350
c.898G > A
Substitution
p.Asp300Asn
Substitution
2B
WRN
Atypical Werner syndrome
277700
#
17875
c.898G > A
Substitution
p.Asp300Asn
Substitution
2B
WRN
Atypical Werner syndrome
277700
#
11767
c.1130G > T
Substitution
p.Arg377Leu
Substitution
2B
AD-SMA
Autosomal dominant spinal
182980
#
muscular dystrophy
11766
c.1477C > T
Substitution
p.Gln493X
Substitution
Tail
AD-SMA
Autosomal dominant spinal
182980
#
muscular dystrophy
9205
c.?
Unknown
p.Glu33Asp
Substitution
1A
—
Axonal neuropathy,
182980
—
muscular dystrophy, cardiac
disease
8994
c.99G > T
Substitution
p.Glu33Asp
Substitution
1A
—
Axonal neuropathy,
—
—
muscular dystrophy, cardiac
disease, leuconychia
12416
c.1621C > T
Substitution
p.Arg541Cys
Substitution
Tail
—
Cardiac arrhythmia
—
—
12399
c.673C > T
Substitution
p.Arg225X
Substitution
L12
CCD
Cardiac conduction defect
115080
#
17808
c.695G > T
Substitution
p.Gly232Val
Substitution
L12
CCD
Cardiac conduction defect
115080
#
13449
c.799T > C
Substitution
p.Tyr267His
Substitution
2B
CCD
Cardiac conduction defect
115080
#
14260
c.178C > G
Substitution
p.Arg60Gly
Substitution
1A
—
Cardiomyopathy with
—
—
advanced AV block and
arrhythmia
14256
c.184C > G
Substitution
p.Arg62Gly
Substitution
1A
—
Cardiomyopathy with
—
—
advanced AV block and
arrhythmia
14018
c.497G > C
Substitution
p.Arg166Pro
Substitution
1B
—
Cardiomyopathy with
—
—
advanced AV block and
arrhythmia
14258
c.575A > T
Substitution
p.Asp192Val
Substitution
1B
—
Cardiomyopathy with
—
—
advanced AV block and
arrhythmia
14007
c.673C > T
Substitution
p.Arg225X
Substitution
L12
—
Cardiomyopathy with
—
—
advanced AV block and
arrhythmia
14015
c.775T > C
Substitution
p.Tyr259His
Substitution
L2
—
Cardiomyopathy with
—
—
advanced AV block and
arrhythmia
14016
c.775T > C
Substitution
p.Tyr259His
Substitution
L2
—
Cardiomyopathy with
—
—
advanced AV block and
arrhythmia
14011
c.815_818delACAAins
Indel
p.Asp272AlafsX208
Frame shift
2B
—
Cardiomyopathy with
—
—
CCAGAC
advanced AV block and
arrhythmia
14012
c.815_818delACAAins
Indel
p.Asp272AlafsX208
Frame shift
2B
—
Cardiomyopathy with
CCAGAC
advanced AV block and
—
—
arrhythmia
11652
c.-
Deletion
p.Met1_Pro4del
Deletion
Head
CMT2
Charcot-Marie-Tooth disease
118210
#
3_12delGCCATGGAGA
type 2
CCCCG
13374
c.1496_1496delC
Deletion
p.Ala499ValfsX141
Frame shift
Tail
CMT2
Charcot-Marie-Tooth disease
118210
#
type 2
17199
c.1910T > C
Substitution
p.Phe637Ser
Substitution
Tail
CMT2
Charcot-Marie-Tooth disease
118210
#
type 2
8840
c.8920 > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
11415
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
11416
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
8997
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
11840
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
11839
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
11838
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
11837
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
12087
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13414
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13415
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13416
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13417
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13418
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13419
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13420
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13421
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13422
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13423
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13424
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13425
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13426
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13427
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13428
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13429
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13430
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13431
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13432
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13433
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13434
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13435
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13436
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
13437
c.892C > T
Substitution
p.Arg298Cys
Substitution
2B
CMT2B1
Charcot-Marie-Tooth disease
605588
#
type 2B1
17886
c.80C > T
Substitution
p.Thr27Ile
Substitution
Head
CFTDM
Congenital fiber type
255310
#
disproportion
18472
c.907T > C
Substitution
p.Ser303Pro
Substitution
2B
CFTDM
Congenital fiber type
255310
#
disproportion
18473
c.907T > C
Substitution
p.Ser303Pro
Substitution
2B
CFTDM
Congenital fiber type
255310
#
disproportion
18474
c.907T > C
Substitution
p.Ser303Pro
Substitution
2B
CFTDM
Congenital fiber type
255310
#
disproportion
18475
c.907T > C
Substitution
p.Ser303Pro
Substitution
2B
CFTDM
Congenital fiber type
255310
#
disproportion
18144
c.91G > A
Substitution
p.Glu31Lys
Substitution
Head
CMD
Congenital muscular
—
—
dystrophy
18148
c.91_93delGAG
Deletion
p.Glu31X
Substitution
Head
CMD
Congenital muscular
—
—
dystrophy
17813
c.93G > C
Substitution
p.Glu31Asp
Substitution
Head
CMD
Congenital muscular
—
—
dystrophy
18146
c.94_96delAAG
Deletion
p.Lys32X
Substitution
1A
CMD
Congenital muscular
—
—
dystrophy
16402
c.104T > C
Substitution
p.Leu35Pro
Substitution
1A
CMD
Congenital muscular
—
—
dystrophy
16287
c.115A > T
Substitution
p.Asn39Tyr
Substitution
1A
CMD
Congenital muscular
—
—
dystrophy
18150
c.117T > G
Substitution
p.Asn39Lys
Substitution
1A
CMD
Congenital muscular
—
—
dystrophy
18153
c.143G > C
Substitution
p.Arg48Pro
Substitution
1A
CMD
Congenital muscular
—
—
dystrophy
18156
c.422T > C
Substitution
p.Leu141Pro
Substitution
1B
CMD
Congenital muscular
—
—
dystrophy
13462
c.745C > T
Substitution
p.Arg249Trp
Substitution
2A
CMD
Congenital muscular
—
—
dystrophy
16404
c.745C > T
Substitution
p.Arg249Trp
Substitution
2A
CMD
Congenital muscular
—
—
dystrophy
17758
c.745C > T
Substitution
p.Arg249Trp
Substitution
2A
CMD
Congenital muscular
—
—
dystrophy
18158
c.745C > T
Substitution
p.Arg249Trp
Substitution
2A
CMD
Congenital muscular
—
—
dystrophy
11816
c.1072G > A
Substitution
p.Glu358Lys
Substitution
2B
CMD
Congenital muscular
—
—
dystrophy
18161
c.1117A > G
Substitution
p.Ile373Val
Substitution
2B
CMD
Congenital muscular
—
—
dystrophy
18166
c.1118-T > A
Substitution
p.Ile373Asn
Substitution
2B
CMD
Congenital muscular
—
—
dystrophy
13460
c.1139-T > C
Substitution
p.Leu380Ser
Substitution
2B
CMD
Congenital muscular
—
—
dystrophy
18169
c.1147G > A
Substitution
p.Glu383Lys
Substitution
2B
CMD
Congenital muscular
—
—
dystrophy
18171
c.1147G > A
Substitution
p.Glu383Lys
Substitution
2B
CMD
Congenital muscular
—
—
dystrophy
18163
c.1151A > G
Substitution
p.Glu384Gly
Substitution
2B
CMD
Congenital muscular
—
—
dystrophy
11817
c.1162C > T
Substitution
p.Arg388Cys
Substitution
Tail
CMD
Congenital muscular
—
—
dystrophy
17473
c.1330_1338dupGAGG
Duplication
p.Glu444_Asp446dup
Duplication
Tail
CMD
Congenital muscular
—
—
TGGAT
dystrophy
11818
c.1368_1370delCAA
Deletion
p.Asn456del
Deletion
Tail
CMD
Congenital muscular
—
—
dystrophy
18238
c.1489-14 1489-
Deletion
p.?
Unknown
Unknown
CMD
Congenital muscular
—
—
7delTTTCTCCT
dystrophy
16951
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
T2D
DIABETES MELLITUS,
125853
#
NONINSULIN-DEPENDENT,
NIDDM
13171
c.?
Unknown
p.Lys260Asn
Substitution
L2
CMD1A
Dilated cardiomyopathy 1A
115200
#
8832
c.16C > T
Substitution
p.Gln6X
Substitution
Head
CMD1A
Dilated cardiomyopathy 1A
115200
#
9397
c.28_29insA
Insertion
p.Thr10AsnfsX31
Frame shift
Head
CMD1A
Dilated cardiomyopathy 1A
115200
#
11627
c.31delC
Deletion
p.Arg11AlafsX85
Frame shift
Head
CMD1A
Dilated cardiomyopathy 1A
115200
#
14254
c.73C > G
Substitution
p.Arg25Gly
Substitution
Head
CMD1A
Dilated cardiomyopathy 1A
115200
#
13155
c.82C > T
Substitution
p.Arg28Trp
Substitution
Head
CMD1A
Dilated cardiomyopathy 1A
115200
#
13563
c.155T > C
Substitution
p.Leu52Pro
Substitution
1A
CMD1A
Dilated cardiomyopathy 1A
115200
#
13379
c.176T > G
Substitution
p.Leu59Arg
Substitution
1A
CMD1A
Dilated cardiomyopathy 1A
115200
#
13378
c.176T > G
Substitution
p.Leu59Arg
Substitution
1A
CMD1A
Dilated cardiomyopathy 1A
115200
#
8748
c.178C > G
Substitution
p.Arg60Gly
Substitution
1A
CMD1A
Dilated cardiomyopathy 1A
115200
#
13192
c.203_208delAGGTGG
Deletion
p.Glu68_Val69del
Deletion
1A
CMD1A
Dilated cardiomyopathy 1A
115200
#
17742
c.232G > A
Substitution
p.Lys78Glu
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
9163
c.244G > A
Substitution
p.Glu82Lys
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
16139
c.244G > A
Substitution
p.Glu82Lys
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
8749
c.254T > G
Substitution
p.Leu85Arg
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
8865
c.266G > T
Substitution
p.Arg89Leu
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
11618
c.266G > T
Substitution
p.Arg89Leu
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13080
c.266G > T
Substitution
p.Arg89Leu
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13157
c.266G > T
Substitution
p.Arg89Leu
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
14030
c.266G > T
Substitution
p.Arg89Leu
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
16966
c.266G > T
Substitution
p.Arg89Leu
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17777
c.266G > T
Substitution
p.Arg89Leu
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13613
c.274C > T
Substitution
p.Leu92Phe
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
8844
c.289A > G
Substitution
p.Lys97Glu
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
11620
c.289A > G
Substitution
p.Lys97Glu
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13159
c.289A > G
Substitution
p.Lys97Glu
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13082
c.302G > C
Substitution
p.Arg101Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
16954
c.302G > C
Substitution
p.Arg101Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
8843
c.331G > T
Substitution
p.Glu111X
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
11619
c.331G > T
Substitution
p.Glu111X
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13199
c.331G > T
Substitution
p.Glu111X
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
16935
c.348_349insG
Insertion
p.Lys117GlufsX10
Frame shift
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13089
c.357-1G > T
Substitution
p.?
Unknown
Unknown
CMD1A
Dilated cardiomyopathy 1A
115200
#
9148
c.394G > C
Substitution
p.Ala132Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17029
c.394G > C
Substitution
p.Ala132Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17028
c.394G > C
Substitution
p.Ala132Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
9297
c.425_426insGGCACT
Insertion
p.Leu141_Asn142insL
Insertion
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
GGAGGCTCTGCTGAA
ysAspLeuAspAlaLeu
Leu
9010
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
11473
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
11474
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
11475
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy lA
115200
#
11476
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
11477
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
9150
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
16162
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
16163
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
16164
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
16165
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17038
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17037
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17036
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17024
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17022
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17021
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17020
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17019
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17018
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17023
c.427T > C
Substitution
p.Ser143Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17661
c.448A > T
Substitution
p.Thr150Ser
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
8879
c.481G > A
Substitution
p.Glu161Lys
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13161
c.481G > A
Substitution
p.Glu161Lys
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13162
c.481G > A
Substitution
p.Glu161Lys
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13163
c.481G > A
Substitution
p.Glu161Lys
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13328
c.481G > A
Substitution
p.Glu161Lys
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13617
c.481G > A
Substitution
p.Glu161Lys
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13091
c.497G > C
Substitution
p.Arg166Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
16968
c.497G > C
Substitution
p.Arg166Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
16969
c.497G > C
Substitution
p.Arg166Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13165
c.548T > C
Substitution
p.Leu183Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
18267
c.563T > G
Substitution
p.Leu188Arg
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
16160
c.565C > T
Substitution
p.Arg189Trp
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
8845
c.568C > T
Substitution
p.Arg190Trp
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
9011
c.568C > T
Substitution
p.Arg190Trp
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
11507
c.568C > T
Substitution
p.Arg190Trp
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
9149
c.568C > T
Substitution
p.Arg190Trp
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
11621
c.568C > T
Substitution
p.Arg190Trp
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
11782
c.568C > T
Substitution
p.Arg190Trp
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
11790
c.568C > T
Substitution
p.Arg190Trp
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13167
c.568C > T
Substitution
p.Arg190Trp
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
16167
c.568C > T
Substitution
p.Arg190Trp
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17034
c.568C > T
Substitution
p.Arg190Trp
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17033
c.568C > T
Substitution
p.Arg190Trp
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17032
c.568C > T
Substitution
p.Arg190Trp
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17031
c.568C > T
Substitution
p.Arg190Trp
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13093
c.569G > A
Substitution
p.Arg190Gln
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13330
c.569G > A
Substitution
p.Arg190Gln
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
16976
c.569G > A
Substitution
p.Arg190Gln
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
9165
c.575A > G
Substitution
p.Asp192Gly
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
14200
c.575A > G
Substitution
p.Asp192Gly
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
8750
c.585C > G
Substitution
p.Asn195Lys
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
12393
c.585C > A
Substitution
p.Asn195Lys
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
11668
c.607G > A
Substitution
p.Glu203Lys
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
16978
c.607G > A
Substitution
p.Glu203Lys
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
8751
c.6084 > G
Substitution
p.Glu203Gly
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13332
c.608A > T
Substitution
p.Glu203Val
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13095
c.629T > G
Substitution
p.Ile210Ser
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
16971
c.629T > G
Substitution
p.Ile210Ser
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
8901
c.640 − 10A > G
Substitution
p.?
Unknown
Unknown
CMD1A
Dilated cardiomyopathy lA
115200
#
8864
c.644T > C
Substitution
p.Leu215Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
16980
c.644T > C
Substitution
p.Leu215Pro
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13169
c.656A > C
Substitution
p.Lys219Thr
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13335
c.656A > C
Substitution
p.Lys219Thr
Substitution
1B
CMD1A
Dilated cardiomyopathy 1A
115200
#
11669
c.673C > T
Substitution
p.Arg225X
Substitution
L12
CMD1A
Dilated cardiomyopathy 1A
115200
#
17607
c.673C > T
Substitution
p.Arg225X
Substitution
L12
CMD1A
Dilated cardiomyopathy 1A
115200
#
17609
c.673C > T
Substitution
p.Arg225X
Substitution
L12
CMD1A
Dilated cardiomyopathy 1A
115200
#
13097
c.700C > T
Substitution
p.Gln234X
Substitution
L12
CMD1A
Dilated cardiomyopathy lA
115200
#
13201
c.736C > T
Substitution
p.Gln246X
Substitution
2A
CMD1A
Dilated cardiomyopathy 1A
115200
#
17786
c.736C > T
Substitution
p.Gln246X
Substitution
2A
CMD1A
Dilated cardiomyopathy 1A
115200
#
17779
c.767T > G
Substitution
p.Val256Gly
Substitution
2A
CMD1A
Dilated cardiomyopathy 1A
115200
#
11809
c.8004 > G
Substitution
p.Tyr267Cys
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13173
c.8004 > G
Substitution
p.Tyr267Cys
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
11810
c.855delG
Deletion
p.Ala287LeufsX191
Frame shift
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
11613
c.908_909delCT
Deletion
p.Ser303CysfsX26
Frame shift
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
12384
c.908_909delCT
Deletion
p.Ser303CysfsX26
Frame shift
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13206
c.936 + 1G > T
Substitution
p.?
Unknown
Unknown
CMD1A
Dilated cardiomyopathy 1A
115200
#
16805
c.937 − 11C > G
Substitution
p.Leu313GlylsX31
Frame shift
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
8846
c.949G > A
Substitution
p.Glu317Lys
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
11617
c.949G > A
Substitution
p.Glu317Lys
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13175
c.949G > A
Substitution
p.Glu317Lys
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13176
c.949G > A
Substitution
p.Glu317Lys
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13621
c.949G > A
Substitution
p.Glu317Lys
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13100
c.952G > A
Substitution
p.Ala318Thr
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
16956
c.952G > A
Substitution
p.Ala318Thr
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
11700
c.959delT
Deletion
p.Leu320fsX160
Frame shift
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
9409
c.959delT
Deletion
p.Leu320fsX160
Frame shift
2B
CMD1A
Dilated cardiomyopathy lA
115200
#
9391
c.959delT
Deletion
p.Leu320fsX160
Frame shift
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
14032
c.959delT
Deletion
p.Leu320fsX160
Frame shift
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
12389
c.961C > T
Substitution
p.Arg321X
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
14077
c.961C > T
Substitution
p.Arg321X
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
14093
c.961C > T
Substitution
p.Arg321X
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17737
c.961C > T
Substitution
p.Arg321X
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17738
c.961C > T
Substitution
p.Arg321X
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
9155
c.976T > A
Substitution
p.Ser326Thr
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
11760
c.[992G > A; =] +
Substitution
p.[Arg331Glu; =] +
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
[=; 1039G > A]
[=; Glu347Lys]
11811
c.992G > C
Substitution
p.Arg331Pro
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
14075
c.992G > A
Substitution
p.Arg331Gln
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17477
c.1003C > T
Substitution
p.Arg335Trp
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
18486
c.1039G > A
Substitution
p.Glu347Lys
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
9012
c.1046G > T
Substitution
p.Arg349Leu
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13561
c.1048G > C
Substitution
p.Ala350Pro
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
14204
c.1057C > A
Substitution
p.Gln353Lys
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
14072
c.1063C > T
Substitution
p.Gln355X
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17475
c.1070A > C
Substitution
p.Asp357Ala
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13116
c.1072G > T
Substitution
p.Glu358X
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
9152
c.1085_1085delT
Deletion
p.Leu363TrpfsX117
Frame shift
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17026
c.1085_1085delT
Deletion
p.Leu362TrpfsX117
Frame shift
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
16157
c.1102_1130dupGCCC
Duplication
p.Lys378ProfsX112
Frame shift
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
TGGACATGGAGATCC
ACGCCTACCG
13102
c.1114delG
Deletion
p.Glu372ArgfsX107
Frame shift
2B
CMD1A
Dilated cardiomyopathy lA
115200
#
8866
c.1130G > A
Substitution
p.Arg377His
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
8869
c.1130G > A
Substitution
p.Arg377His
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
8880
c.1130G > A
Substitution
p.Arg377His
Substitution
2B
CMD1A
Dilated cardiomyopathy lA
115200
#
9160
c.1130G > A
Substitution
p.Arg377His
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
9162
c.1130G > A
Substitution
p.Arg377His
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
12330
c.1130G > A
Substitution
p.Arg377His
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13178
c.1130G > T
Substitution
p.Arg377Leu
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13623
c.1130G > A
Substitution
p.Arg377His
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
14034
c.1130G > A
Substitution
p.Arg377His
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17664
c.1150G > T
Substitution
p.Glu384X
Substitution
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
17482
C.1157 + 1G > T
Substitution
p.Arg386SerfsX21
Frame shift
2B
CMD1A
Dilated cardiomyopathy 1A
115200
#
13104
c.1163G > A
Substitution
p.Arg388His
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
16958
c.1163G > A
Substitution
p.Arg388His
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
13106
c.1195C > T
Substitution
p.Arg399Cys
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
16960
c.1195C > T
Substitution
p.Arg399Cys
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
17782
c.1197_1240delTGGCC
Deletion
p.Gly400ArgfsX11
Frame shift
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
GTGCTTCCTCTCACTC
ATCCCAGACACAGGG
TGGGGGCA
17789
c.1292C > G
Substitution
p.Ser431X
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
14082
c.1294C > T
Substitution
p.Gln432X
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
14084
c.1294C > T
Substitution
p.Gln432X
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
14085
c.1294C > T
Substitution
p.Gln432X
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
14086
c.1294C > T
Substitution
p.Gln432X
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
13108
c.1307_1308insGCAC
Insertion
p.Ser437HisfsX1
Frame shift
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
16962
c.1307_1308insGCAC
Insertion
p.Ser437HisfsX1
Frame shift
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
14088
c.1318G > A
Substitution
p.Val440Met
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
14090
c.1318G > A
Substitution
p.Val440Met
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
14091
c.1318G > A
Substitution
p.Val440Met
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
11812
c.1370delA
Deletion
p.Lys457SerfsX21
Frame shift
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
12397
c.1380 + 1G > A
Substitution
p.?
Unknown
Unknown
CMD1A
Dilated cardiomyopathy 1A
115200
#
9413
c.1397_1397delA
Deletion
p.Asn466IlefsX14
Frame shift
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
9161
c.1397_1397delA
Deletion
p.Asn466IlefsX14
Frame shift
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
13110
c.1412G > A
Substitution
p.Arg471His
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
16973
c.1412G > A
Substitution
p.Arg471His
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
13112
c.1424_1425insAGA
Insertion
p.Gly474_Asp475ins
Insertion
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
Glu
11508
c.1443C > G
Substitution
p.Tyr481X
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
17791
c.1443C > G
Substitution
p.Tyr481X
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
17480
c.1489 − 1G > T
Substitution
p.Ile497_Glu536del
Deletion
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
13180
c.1492T > A
Substitution
p.Trp498Arg
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
9153
c.1493_1493delG
Deletion
p.Ala499LeufsX47
Frame shift
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
12395
c.1512_1513insAG
Insertion
p.Thr505ArgfsX44
Frame shift
Tail
CMD1A
Dilated cardiomyopathy lA
115200
#
17784
c.1526_1527insC
Insertion
p.Thr510TyrfsX42
Frame shift
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
17797
c.1526_1527insA
Insertion
p.Thr510TyrfsX42
Frame shift
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
17486
c.1549C > T
Substitution
p.Gln517X
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
17484
c.1560G > A
Substitution
p.Trp520X
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
13625
c.1567G > A
Substitution
p.Gly523Arg
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
11624
c.1579_1580insCTGC
Insertion
p.Arg527ProlsX26
Frame shift
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
17558
c.1583C > T
Substitution
p.Thr528Met
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
9307
c.1621C > T
Substitution
p.Arg541Cys
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
9283
c.1621C > A
Substitution
p.Arg541Ser
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
9151
c.1621C > A
Substitution
p.Arg541Ser
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
13592
c.1621C > T
Substitution
p.Arg541Cys
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
14202
c.1621C > A
Substitution
p.Arg541Ser
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
16472
c.1621C > G
Substitution
p.Arg541Gly
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
17774
c.1621C > T
Substitution
p.Arg541Cys
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
17775
c.1621C > T
Substitution
p.Arg541Cys
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
13559
c.1622G > A
Substitution
p.Arg541His
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
9392
c.1718C > T
Substitution
p.Ser573Leu
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
13182
c.1718C > T
Substitution
p.Ser573Leu
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
13183
c.1718C > T
Substitution
p.Ser573Leu
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
14038
c.1718C > T
Substitution
p.Ser573Leu
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
17667
c.1879C > T
Substitution
p.Arg624Cys
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
13565
c.1904G > A
Substitution
p.Gly635Asp
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
8833
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
9018
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
11500
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
11501
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
11502
c.1930C > A
Substitution
p.Arg644His
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
13185
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
13337
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
14080
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
13114
c.1960C > T
Substitution
p.Arg654X
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
16964
c.1960C > T
Substitution
p.Arg654X
Substitution
Tail
CMD1A
Dilated cardiomyopathy 1A
115200
#
11788
c.?
Unknown
p.Tyr481X
Substitution
Tail
DCM-CD
Dilated cardiomyopathy with
—
—
conduction system defects
17886
c.80C > T
Substitution
p.Thr27Ile
Substitution
Head
DCM-CD
Dilated cardiomyopathy with
—
—
conduction system defects
13486
c.106C > T
Substitution
p.Gln36X
Substitution
1A
DCM-CD
Dilated cardiomyopathy with
—
—
conduction system defects
11792
c.158A > T
Substitution
p.Glu53Val
Substitution
1A
DCM-CD
Dilated cardiomyopathy with
—
—
conduction system defects
18301
c.175C > G
Substitution
p.Leu59Val
Substitution
1A
DCM-CD
Dilated cardiomyopathy with
—
—
conduction system defects
11791
c.481G > A
Substitution
p.Glu161Lys
Substitution
1B
DCM-CD
Dilated cardiomyopathy with
—
—
conduction system defects
13204
c.514 − 1G > A
Substitution
p.?
Unknown
Unknown
DCM-CD
Dilated cardiomyopathy with
—
—
conduction system defects
11793
c.556G > A
Substitution
p.Glu186Lys
Substitution
1B
DCM-CD
Dilated cardiomyopathy with
—
—
conduction system defects
11789
c.568C > T
Substitution
p.Arg190Trp
Substitution
1B
DCM-CD
Dilated cardiomyopathy with
—
—
conduction system defects
11787
c.575A > G
Substitution
p.Asp192Gly
Substitution
1B
DCM-CD
Dilated cardiomyopathy with
—
—
conduction system defects
18470
c.683A > T
Substitution
p.Glu228Val
Substitution
L12
DCM-CD
Dilated cardiomyopathy with
—
—
conduction system defects
17868
c.871G > A
Substitution
p.Glu291Lys
Substitution
2B
DCM-CD
Dilated cardiomyopathy with
—
—
conduction system defects
17870
c.949G > A
Substitution
p.Glu317Lys
Substitution
2B
DCM-CD
Dilated cardiomyopathy with
—
—
conduction system defects
13078
c.1069G > C
Substitution
p.Asp357His
Substitution
2B
DCM-CD
Dilated cardiomyopathy with
—
—
conduction system defects
13208
C.1157 + 1G > A
Substitution
p.?
Unknown
Unknown
DCM-CD
Dilated cardiomyopathy with
—
—
conduction system defects
18490
c.1412G > A
Substitution
p.Arg471His
Substitution
Tail
DCM-CD
Dilated cardiomyopathy with
—
—
conduction system defects
11814
c.1526_1527insC
Insertion
p.Thr510TyrfsX42
Frame shift
Tail
DCM-CD
Dilated cardiomyopathy with
—
—
conduction system defects
9008
c.1621C > T
Substitution
p.Arg541Cys
Substitution
Tail
DCM-CD
Dilated cardiomyopathy with
—
—
conduction system defects
17578
c.1711C > A
Substitution
p.=
Silent
Not
DCM-CD
Dilated cardiomyopathy with
—
—
affected
conduction system defects
17880
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
DCM-CD
Dilated cardiomyopathy with
—
—
conduction system defects
18305
c.1774G > A
Substitution
p.Gly592Arg
Substitution
Tail
DAPJ
Distal acroosteolysis,
—
—
poikiloderma and joint stiffness
13581
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
—
Distal motor neuropathy
—
—
17813
c.93G > C
Substitution
p.Glu31Asp
Substitution
Head
—
Dropped head syndrome
—
—
9020
c.94_96delAAG
Deletion
p.Lys32del
Deletion
1A
—
Dropped head syndrome
—
—
13476
c.116A > G
Substitution
p.Asn39Ser
Substitution
1A
—
Dropped head syndrome
—
—
13466
c.149G > C
Substitution
p.Arg50Pro
Substitution
1A
—
Dropped head syndrome
—
—
13464
c.745C > T
Substitution
p.Arg249Trp
Substitution
2A
—
Dropped head syndrome
—
—
13470
c.905T > C
Substitution
pleu302Pro
Substitution
2B
—
Dropped head syndrome
—
—
13468
c.1072G > A
Substitution
p.Glu358Lys
Substitution
2B
—
Dropped head syndrome
—
—
13482
c.1072G > A
Substitution
p.Glu358Lys
Substitution
2B
—
Dropped head syndrome
—
—
13484
c.1072G > A
Substitution
p.Glu358Lys
Substitution
2B
—
Dropped head syndrome
—
—
13474
c.1358G > C
Substitution
p.Arg453Pro
Substitution
Tail
—
Dropped head syndrome
—
—
13472
c.1364G > C
Substitution
p.Arg455Pro
Substitution
Tail
—
Dropped head syndrome
—
—
13478
c.1366A > G
Substitution
p.Asn456Asp
Substitution
Tail
—
Dropped head syndrome
—
—
13480
c.1381-2A > G
Substitution
p.?
Unknown
Unknown
—
Dropped head syndrome
—
—
11652
c.-
Deletion
p.Met1_Pro4del
Deletion
Head
EDMD2
Emery-Dreifuss muscular
181350
#
3_12delGCCATGGAGA
dystrophy, autosomal
CCCCG
dominant
8743
c.16C > T
Substitution
p.Gln6X
Substitution
Head
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8796
c.16C > T
Substitution
p.Gln6X
Substitution
Head
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11746
c.31delC
Deletion
p.Arg11AlafsX85
Frame shift
Head
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9262
c.73C > G
Substitution
p.Arg25Gly
Substitution
Head
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9376
c.74G > C
Substitution
p.Arg25Pro
Substitution
Head
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9177
c.94_96delAAG
Deletion
p.Lys32del
Deletion
1A
EDMD2
Emery-Dreifuss muscular
181350
#
EDMD2 dystrophy, autosomal
dominant
8998
c.94_96delAAG
Deletion
p.Lys32del
Deletion
1A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9014
c.94_96delAAG
Deletion
p.Lys32del
Deletion
1A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13594
c.98A > G
Substitution
p.Glu33Gly
Substitution
1A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11747
c.99G > C
Substitution
p.Glu33Asp
Substitution
1A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8999
c.103C > G
Substitution
p.Leu35Val
Substitution
1A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11748
c.116A > G
Substitution
p.Asn39Ser
Substitution
1A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16329
c.116A > G
Substitution
p.Asn39Ser
Substitution
1A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
18174
c.116A > G
Substitution
p.Asn39Ser
Substitution
1A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9377
c.127G > A
Substitution
p.Ala43Thr
Substitution
1A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8797
c.134A > G
Substitution
p.Tyr45Cys
Substitution
1A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16331
c.134A > G
Substitution
p.Tyr45Cys
Substitution
1A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13605
c.136A > G
Substitution
p.Ile46Val
Substitution
1A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13545
c.139G > C
Substitution
p.Asp47His
Substitution
1A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9378
c.148C > A
Substitution
p.Arg50Ser
Substitution
1A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8798
c.149G > C
Substitution
p.Arg50Pro
Substitution
1A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8799
c.188T > G
Substitution
p.Ile63Ser
Substitution
1A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9175
c.188T > A
Substitution
p.Ile63Asn
Substitution
1A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11633
c.188T > A
Substitution
p.Ile63Asn
Substitution
1A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13192
c.203_208delAGGTGG
Deletion
p.Glu68_Val69del
Deletion
1A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
12477
c.265C > T
Substitution
p.Arg89Cys
Substitution
1B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13596
c.265C > T
Substitution
p.Arg89Cys
Substitution
1B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
17193
c.266G > T
Substitution
p.Arg89Leu
Substitution
1B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9410
c.334_336delGAG
Deletion
p.Glu112del
Deletion
1B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9000
c.334_336delGAG
Deletion
p.Glu112del
Deletion
1B
EDMD2
Emery-Dreifuss muscular
181350
#
EDMD2 dystrophy, autosomal
dominant
16291
c.357C > T
Substitution
p.=
Silent
Not
EDMD2
Emery-Dreifuss muscular
181350
#
affected
dystrophy, autosomal
dominant
17182
c.367_369delAAG
Deletion
p.Lys123del
Deletion
1B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9379
c.398G > C
Substitution
p.Arg133Pro
Substitution
1B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11632
c.419T > C
Substitution
p.Leu140Pro
Substitution
1B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9015
c.428C > T
Substitution
p.Ser143Phe
Substitution
1B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
12609
c.428C > T
Substitution
p.Ser143Phe
Substitution
1B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8795
c.448A > C
Substitution
p.Thr150Pro
Substitution
1B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16334
c.448A > C
Substitution
p.Thr150Pro
Substitution
1B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13453
c.485T > C
Substitution
p.Leu162Pro
Substitution
1B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16396
c.566_567delGGinsCC
Indel
p.Arg189Pro
Substitution
1B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16299
c.568_570dupCGG
Duplication
p.Arg190dup
Duplication
1B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9380
c.588_596delGCTGCA
Deletion
p.Arg196_Thr199delin
Indel
1B
EDMD2
Emery-Dreifuss muscular
181350
#
GAC
sSer
dystrophy, autosomal
dominant
16301
c.618C > G
Substitution
p.Phe206Leu
Substitution
1B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
12382
c.625delA
Deletion
p.Asn209ThrfsX271
Frame shift
1B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8800
c.665A > C
Substitution
p.His222Pro
Substitution
L12
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13547
c.694G > C
Substitution
p.Gly232Arg
Substitution
L12
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8801
c.695G > A
Substitution
p.Gly232Glu
Substitution
L12
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
12485
c.695G > A
Substitution
p.Gly232Glu
Substitution
L12
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9001
c.743T > C
Substitution
p.Leu248Pro
Substitution
2A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16336
c.745C > T
Substitution
p.Arg249Trp
Substitution
2A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16337
c.745C > T
Substitution
p.Arg249Trp
Substitution
2A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16338
c.745C > T
Substitution
p.Arg249Trp
Substitution
2A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16342
c.[745C > T; 1930C > T]
Substitution
p.[Arg249Trp;
Substitution
2A, Tail
EDMD2
Emery-Dreifuss muscular
181350
#
Arg644Cys]
dystrophy, autosomal
dominant
8783
c.746G > A
Substitution
p.Arg249Gln
Substitution
2A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11380
c.746G > A
Substitution
p.Arg249Gln
Substitution
2A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11381
c.746G > A
Substitution
p.Arg249Gln
Substitution
2A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8802
c.746G > A
Substitution
p.Arg249Gln
Substitution
2A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9002
c.746G > A
Substitution
p.Arg249Gln
Substitution
2A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9023
c.746G > A
Substitution
p.Arg249Gln
Substitution
2A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11631
c.746G > A
Substitution
p.Arg249Gln
Substitution
2A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11749
c.746G > A
Substitution
p.Arg249Gln
Substitution
2A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11750
c.746G > A
Substitution
p.Arg249Gln
Substitution
2A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13552
c.746G > A
Substitution
p.Arg249Gln
Substitution
2A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16344
c.746G > A
Substitution
p.Arg249Gln
Substitution
2A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16345
c.746G > A
Substitution
p.Arg249Gln
Substitution
2A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
18176
c.746G > A
Substitution
p.Arg249Gln
Substitution
2A
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11751
c.775T > G
Substitution
p.Tyr259Asp
Substitution
L2
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9640
c.781_783delAAG
Deletion
p.Lys261del
Deletion
L2
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9411
c.781_783delAAG
Deletion
plys261del
Deletion
L2
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9381
c.781_783delAAG
Deletion
plys261del
Deletion
L2
EDMD2
Emery-Dreifuss muscular
181350
#
EDMD2 dystrophy, autosomal
dominant
13584
c.781_783delAAGinsG
Indel
p. Lys261delinsValGlu
Indel
L2
EDMD2
Emery-Dreifuss muscular
181350
#
TGGAGCAGTATAAGA
GlnTyrLysLys
dystrophy, autosomal
AA
dominant
16149
c.788T > C
Substitution
p.Leu263Pro
Substitution
L2
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
17403
c.788T > C
Substitution
p.Leu263Pro
Substitution
L2
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
17751
c.788T > C
Substitution
p.Leu263Pro
Substitution
L2
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
17405
c.799T > C
Substitution
p.Tyr267His
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9003
c.800A > G
Substitution
p.Tyr267Cys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11809
c.800A > G
Substitution
p.Tyr267Cys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13173
c.800A > G
Substitution
p.Tyr267Cys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16303
c.802T > C
Substitution
p.Ser268Pro
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16305
c.810G > A
Substitution
p.=
Silent
Not
EDMD2
Emery-Dreifuss muscular
181350
#
affected
dystrophy, autosomal
dominant
16307
c.810G > A
Substitution
p.=
Silent
Not
EDMD2
Emery-Dreifuss muscular
181350
#
affected
dystrophy, autosomal
dominant
16309
c.810 + 1G > A
Substitution
p.?
Unknown
Unknown
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16347
c.812T > C
Substitution
p.Leu271Pro
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
18186
c.832G > C
Substitution
p.Ala278Pro
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8803
c.881A > C
Substitution
p.Gln294Pro
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16349
c.881A > C
Substitution
p.Gln294Pro
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16311
c.883T > C
Substitution
p.Ser295Pro
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8863
c.907T > C
Substitution
p.Ser303Pro
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16351
c.907T > C
Substitution
p.Ser303Pro
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8786
c.1007G > A
Substitution
p.Arg336Gln
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16314
c.1064_1066delAGC
Deletion
p.Gln355del
Deletion
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8804
c.1072G > A
Substitution
p.Glu358Lys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11388
c.1072G > A
Substitution
p.Glu358Lys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11816
c.1072G > A
Substitution
p.Glu358Lys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9167
c.1072G > A
Substitution
p.Glu358Lys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11478
c.1072G > A
Substitution
p.Glu358Lys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11479
c.1072G > A
Substitution
p.Glu358Lys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11480
c.1072G > A
Substitution
p.Glu358Lys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11752
c.1072G > A
Substitution
p.Glu358Lys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
12479
c.1072G > A
Substitution
p.Glu358Lys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
12483
c.1072G > A
Substitution
p.Glu358Lys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13598
c.1072G > A
Substitution
p.Glu358Lys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13603
c.1072G > A
Substitution
p.Glu358Lys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16353
c.1072G > A
Substitution
p.Glu358Lys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16354
c.1072G > A
Substitution
p.Glu358Lys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16355
c.1072G > A
Substitution
p.Glu358Lys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16316
c.1081G > A
Substitution
p.Glu361Lys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8806
c.1112T > A
Substitution
p.Met371Lys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
18184
c.1124C > G
Substitution
p.Ala375Gly
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11634
c.1130G > T
Substitution
p.Arg377Leu
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11693
c.1130G > A
Substitution
p.Arg377His
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
12405
c.1130G > A
Substitution
p.Arg377His
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
18223
c.1130G > A
Substitution
p.Arg377His
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
12474
c.1142A > C
Substitution
p.Glu381Ala
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11629
c.1157G > A
Substitution
p.Arg386Lys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13653
c.1157G > T
Substitution
p.Arg386Met
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16357
c.1157G > A
Substitution
p.Arg386Lys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16358
c.1157G > A
Substitution
p.Arg386Lys
Substitution
2B
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16319
c.1158-2A?G
Substitution
p.?
Unknown
Unknown
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11817
c.1162C > T
Substitution
p.Arg388Cys
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9281
c.1187A > G
Substitution
p.Gln396Arg
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8862
c.1201C > T
Substitution
p.Arg401Cys
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9176
c.1201C > T
Substitution
p.Arg401Cys
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9302
c.1201C > T
Substitution
p.Arg401Cys
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9004
c.1337A > T
Substitution
p.Asp446Val
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16321
c.1346G > A
Substitution
p.Gly449Asp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
18192
c.1346G?T
Substitution
p.Gly449Val
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8744
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8787
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11382
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11383
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11384
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8807
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11389
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11390
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8836
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9005
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9304
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11614
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11753
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
12481
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13590
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13607
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13609
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13611
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16363
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16364
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16365
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16366
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16367
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16368
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16323
c.1361T > C
Substitution
p.Leu454Pro
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9513
c.1367A > T
Substitution
p.Asn456Ile
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8810
c.1368C > A
Substitution
p.Asn456Lys
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11818
c.1368_1370delCAA
Deletion
p.Asn456del
Deletion
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
17446
c.1368_1370delCAA
Deletion
p.Asn456del
Deletion
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
17447
c.1368_1370delCAA
Deletion
p.Asn456del
Deletion
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16370
c.1381-2A > G
Substitution
p.?
Unknown
Unknown
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16372
c.[1381-1G > T;
Substitution
p.?; Asp461Tyr
Unknown,
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
1381G > T]
Substitution
dystrophy, autosomal
dominant
16325
c.1399T > C
Substitution
p.Trp467Arg
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8791
c.1406T > C
Substitution
p.Ile469Thr
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
18194
c.1466T > C
Substitution
p.Leu489Pro
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16327
c.1488 + 1G > A
Substitution
p.?
Unknown
Unknown
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13180
c.1492T > A
Substitution
p.Trp498Arg
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16375
c.1526dupC
Duplication
p.Thr510TyrfsX42
Frame shift
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
18188
c.1540T > A
Substitution
p.Trp514Arg
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
18190
c.1540T > A
Substitution
p.Trp514Arg
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
12434
c.1558T > G
Substitution
p.Trp520Gly
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
18179
c.1558T > C
Substitution
p.Trp520Arg
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8811
c.1559G > C
Substitution
p.Trp520Ser
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8745
c.1580G > C
Substitution
p.Arg527Pro
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11344
c.1580G > C
Substitution
p.Arg527Pro
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8792
c.1580G > C
Substitution
p.Arg527Pro
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8812
c.1580G > C
Substitution
p.Arg527Pro
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11391
c.1580G > C
Substitution
p.Arg527Pro
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8838
c.1580G > C
Substitution
p.Arg527Pro
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8859
c.1580G > C
Substitution
p.Arg527Pro
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11423
c.1580G > C
Substitution
p.Arg527Pro
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11630
c.1580G > C
Substitution
p.Arg527Pro
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11754
c.1580G > C
Substitution
p.Arg527Pro
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16377
c.1580G > C
Substitution
p.Arg527Pro
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
18182
c.1580G > C
Substitution
p.Arg527Pro
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8793
c.1583C > A
Substitution
p.Thr528Lys
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8814
c.1583C > A
Substitution
p.Thr528Lys
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
11392
c.1583C > A
Substitution
p.Thr528Lys
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9006
c.1583C > G
Substitution
p.Thr528Arg
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16379
c.1583C > A
Substitution
p.Thr528Lys
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16381
c.1583C > G
Substitution
p.Thr528Arg
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16382
c.1583C > G
Substitution
p.Thr528Arg
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16407
c.1583C > G
Substitution
p.Thr528Arg
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
18196
c.1583C > G
Substitution
p.Thr528Arg
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
18297
c.1588C > T
Substitution
p.Leu530Phe
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
8747
c.1589T > C
Substitution
p.Leu530Pro
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16384
c.1621C > A
Substitution
p.Arg541Ser
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9007
c.1622G > A
Substitution
p.Arg541His
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16387
c.1622G > C
Substitution
p.Arg541Pro
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13588
c.1633C > T
Substitution
p.Arg545Cys
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16389
c.1804G > A
Substitution
p.Arg541Pro
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9383
c.1871G > A
Substitution
p.Arg624His
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9154
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13570
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13571
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13572
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
13573
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
16392
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
EDMD2
Emery-Dreifuss muscular
181350
#
dystrophy, autosomal
dominant
9523
c.664C > T
Substitution
p.His222Tyr
Substitution
L12
EDMD3
Emery-Dreifuss muscular
604929
#
dystrophy, autosomal
recessive
17407
c.674G > A
Substitution
p.Arg225Gln
Substitution
L12
EDMD3
Emery-Dreifuss muscular
604929
#
dystrophy, autosomal
recessive
17573
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
EDMD3
Emery-Dreifuss muscular
604929
#
dystrophy, autosomal
recessive
13319
c.1580G > C
Substitution
p.Arg527Pro
Substitution
Tail
EDMD3
Emery-Dreifuss muscular
604929
#
dystrophy, autosomal
recessive
18345
c.?
Unknown
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
8847
c.82C > T
Substitution
p.Arg28Trp
Substitution
Head
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
18291
c.139G > A
Substitution
p.Asp47Asn
Substitution
1A
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
8861
c.178C > G
Substitution
p.Arg60Gly
Substitution
1A
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
14260
c.178C > G
Substitution
p.Arg60Gly
Substitution
1A
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
8848
c.184C > G
Substitution
p.Arg62Gly
Substitution
1A
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
14256
c.184C > G
Substitution
p.Arg62Gly
Substitution
1A
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
12100
c.398G > T
Substitution
p.Arg133Leu
Substitution
1B
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
14258
c.575A > T
Substitution
p.Asp192Val
Substitution
1B
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
17624
c.667G > A
Substitution
p.Glu223Lys
Substitution
L12
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11775
c.688G > A
Substitution
p.Asp230Asn
Substitution
L12
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11776
c.1195C > T
Substitution
p.Arg399Cys
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
17066
c.1232G > A
Substitution
p.Gly411Asp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
12096
c.1315C > T
Substitution
pArg439Cys
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
17208
c.1315C > T
Substitution
p.Arg439Cys
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11542
c.[1318G > A; =] +
Substitution
p.[Val440Met; =] +
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
[=; 1445G > A]
[=; Arg482Gln]
(Dunnigan type)
8781
c.1394G > A
Substitution
p.Gly465Asp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
12601
c.14110 > G
Substitution
p.Arg471Gly
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
8754
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11358
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11359
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11360
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11361
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11362
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11363
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11364
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11365
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
8773
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11374
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11375
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11376
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11377
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11378
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11379
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
8816
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11393
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11394
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11395
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11396
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11397
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
8834
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
9213
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
9156
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11543
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11544
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11662
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11692
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11699
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
12101
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
12102
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
12103
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
12104
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
12105
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
14066
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
16433
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
16434
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
16682
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
17866
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
17195
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
17464
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
17465
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
17466
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
17467
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
17733
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
17735
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
17744
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
17936
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
18484
c.1444C > T
Substitution
p.Arg482Trp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
8753
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11354
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11355
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11356
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11357
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
8763
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
8764
c.1445G > T
Substitution
p.Arg482Leu
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
8768
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11370
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11371
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11372
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11373
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
8822
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
8868
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
9305
c.1445G > T
Substitution
p.Arg482Leu
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11663
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11667
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
12098
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
12387
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Unknown
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
12418
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Unknown
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
12423
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
13150
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
13367
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
16285
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
16436
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
16437
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
16438
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
16439
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
16440
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
18138
c.1445G > A
Substitution
p.Arg482Gln
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
8765
c.1458G > C
Substitution
p.Lys486Asn
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11366
c.1458G > C
Substitution
p.Lys486Asn
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
8767
c.1458G > T
Substitution
p.Lys486Asn
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
9319
c.1488 + 5G > C
Substitution
p.Ile497_Met664delins
Indel
Tail
FPLD2
Familial partial lipodystrophy
151660
#
ValThrGlyArgAlaLeuGl
(Dunnigan type)
yThrLeuGlyArgProTrpV
alAlaMetGlyAlaLeuGly
X
17455
c.1683G > C
Substitution
p.=
Silent
Not
FPLD2
Familial partial lipodystrophy
151660
#
affected
(Dunnigan type)
11777
c.1718C > T
Substitution
p.Ser573Leu
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
8780
c.1745G > A
Substitution
p.Arg582His
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
16429
c.1745G > A
Substitution
p.Arg582His
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
16430
c.1745G > A
Substitution
p.Arg582His
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
16431
c.1745G > A
Substitution
p.Arg582His
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
9452
c.1751G > A
Substitution
p.Arg584His
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
11545
c.1751G > A
Substitution
p.Arg584His
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
12429
c.1772G > T
Substitution
p.Arg156Cys
Substitution
1B
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
17068
c.1892G > A
Substitution
p.Gly631Asp
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
13575
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
13576
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
13579
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
17461
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
FPLD2
Familial partial lipodystrophy
151660
#
(Dunnigan type)
12601
c.1411C > G
Substitution
p.Arg471Gly
Substitution
Tail
FPLD1
Familial partial lipodystrophy
608600
%
(Köbberling)
11482
c.[1583C > T; =] +
Substitution
p.[Thr528Met; =] +
Substitution
Tail
FPLD1
Familial partial lipodystrophy
608600
%
[=; 1748C > T]
[=; Ser583Leu
(Köbberling)
9183
c.1748C > T
Substitution
p.Ser583Leu
Substitution
Tail
FPLD1
Familial partial lipodystrophy
608600
%
(Köbberling)
11481
c.1748C > T
Substitution
p.Ser583Leu
Substitution
Tail
FPLD1
Familial partial lipodystrophy
608600
%
(Köbberling)
13490
c.29C > T
Substitution
p.Thr10Ile
Substitution
Head
—
Generalized lipoatrophy
—
—
syndrome
8867
c.398G > T
Substitution
p.Arg133Leu
Substitution
1B
—
Generalized lipoatrophy
—
—
syndrome
17762
c.1609-12T > G
Substitution
p.Glu536fsX14
Frame shift
Tail
HSS
Hallermann-Streiff syndrome
234100
%
17260
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
HSS
Hallermann-Streiff syndrome
234100
%
13153
c.1609-12T > G
Substitution
p.Glu536fsX14
Frame shift
Tail
—
Heart-hand syndrome,
610140
%
Slovenian Type
14097
c.11C > G
Substitution
p.Pro4Arg
Substitution
Head
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
14112
c.11C > G
Substitution
p.Pro4Arg
Substitution
Head
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
9172
c.29C > T
Substitution
p.Thr10Ile
Substitution
Head
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
14108
c.29C > T
Substitution
p.Thr10Ile
Substitution
Head
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
14110
c.29C > T
Substitution
p.Thr10Ile
Substitution
Head
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
14101
c.331G > T
Substitution
p.Glu111Lys
Substitution
1B
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
14095
c.406G > C
Substitution
p.Asp136His
Substitution
1B
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
17070
c.412G > A
Substitution
p.Glu138Lys
Substitution
1B
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
9016
c.428C > T
Substitution
p.Ser143Phe
Substitution
1B
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
12609
c.428C > T
Substitution
p.Ser143Phe
Substitution
1B
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
8871
c.433G > A
Substitution
p.Glu145Lys
Substitution
1B
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
14106
c.475G > A
Substitution
p.Glu159Lys
Substitution
1B
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
17626
c.899A > G
Substitution
p.Asp300Gly
Substitution
2B
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
17903
c.917T > G
Substitution
pi
Substitution
2B
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
14114
c.1303C > T
Substitution
p.Arg435Cys
Substitution
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
9393
c.1411C > T
Substitution
p.Arg471Cys
Substitution
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
12615
c.1411C > T
Substitution
p.Arg471Cys
Substitution
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
9394
c.1579C?T
Substitution
p.Arg527Cys
Substitution
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
13119
c.1579C
Substitution
p.Arg527Cys
Substitution
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
13651
c.1579C > T
Substitution
p.Arg527Cys
Substitution
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
11671
c.[1583C > T; =] +
Substitution
p.[Thr528Met; =] +
Substitution
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
[=; 1619T > C]
[=; Met540Thr]
syndrome
9013
c.1626G > C
Substitution
p.Lys542Asn
Substitution
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
9173
c.1733A > T
Substitution
p.Glu578Val
Substitution
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
14099
c.1762-T > C
Substitution
p.Cys588Arg
Substitution
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
14104
c.1762-T > C
Substitution
p.Cys588Arg
Substitution
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
11785
c.1821G > A
Substitution
p.=
Silent
Not
HGPS
Hutchinson-Gilford progeria
176670
#
affected
syndrome
9527
c.1822G > A
Substitution
p.Gly608Ser
Substitution
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
8876
c.1822G > A
Substitution
p.Gly608Ser
Substitution
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
9396
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
9398
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
11428
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
11429
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
11430
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
11431
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
11432
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
11433
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
11434
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
11435
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
11436
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
11437
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
11438
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
11439
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
11440
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
11441
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
11442
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
11443
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
11444
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
9395
c.1824C > T
Substitution
p.?
Unknown
Unknown
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
11449
c.1824C > T
Substitution
p.?
Unknown
Unknown
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
11450
c.1824C > T
Substitution
p.?
Unknown
Unknown
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
11451
c.1824C > T
Substitution
p.?
Unknown
Unknown
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
11452
c.1824C > T
Substitution
p.?
Unknown
Unknown
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
9171
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
9017
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
9019
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
12074
c.1824C > T
Substitution
p.[=,
Silent,
Not
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
affected
syndrome
12075
c.1824C > T
Substitution
p.[=,
Silent,
Not
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
affected
syndrome
12076
c.1824C > T
Substitution
p.[=,
Silent,
Not
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
affected
syndrome
12085
c.1824C > T
Substitution
p.[=,
Silent,
Not
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
affected
syndrome
12354
c.1824C > T
Substitution
p.=
Silent
Not
HGPS
Hutchinson-Gilford progeria
176670
#
affected
syndrome
13533
c.1824C > T
Substitution
p.[=,
Silent,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val607_Gln656del]
Deletion
syndrome
17393
c.1824C > T
Substitution
p.=
Silent
Not
HGPS
Hutchinson-Gilford progeria
176670
#
affected
syndrome
17395
c.1824C > T
Substitution
p.=
Silent
Not
HGPS
Hutchinson-Gilford progeria
176670
#
affected
syndrome
17564
c.1824C > T
Substitution
p.=
Silent
Not
HGPS
Hutchinson-Gilford progeria
176670
#
affected
syndrome
17729
c.1824C > T
Substitution
p.=
Silent
Not
HGPS
Hutchinson-Gilford progeria
176670
#
affected
syndrome
17818
c.1824C > T
Substitution
p.=
Silent
Not
HGPS
Hutchinson-Gilford progeria
176670
#
affected
syndrome
9009
c.1868C > G
Substitution
p.[Thr623Ser,
Deletion,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val622_Gln656del]
Substitution
syndrome
12457
c.1868C > G
Substitution
p.[Thr623Ser,
Deletion,
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
Val622_Gln656del]
Substitution
syndrome
9174
c.1930C > T
Substitution
p.Arg644Cys
Substitution
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
11615
c.1960C > T
Substitution
p.Arg654X
Substitution
Tail
HGPS
Hutchinson-Gilford progeria
176670
%
syndrome
17159
c.1968G > A
Substitution
p.=
Silent
Not
HGPS
Hutchinson-Gilford progeria
176670
#
affected
syndrome
11784
c.1968 + 1G > A
Substitution
p.Val607_Gln656del
Deletion
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
17161
c.1968 + 5G > A
Substitution
p.Val607_Gln656del
Deletion
Tail
HGPS
Hutchinson-Gilford progeria
176670
#
syndrome
12615
c.1411C > T
Substitution
p.Arg471Cys
Substitution
Tail
—
Lamin-related rigid spine
—
—
muscular dystrophy
17426
c.31delC
Deletion
p.Arg11AlafsX85
Frame shift
Head
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
14254
c.73C > G
Substitution
p.Arg25Gly
Substitution
Head
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
17886
c.80C > T
Substitution
p.Thr27Ile
Substitution
Head
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
18488
c.80C?T
Substitution
p.Thr27Ile
Substitution
Head
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
17428
c.99G > C
Substitution
p.Glu33Asp
Substitution
1A
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
17429
c.99G > C
Substitution
p.Glu33Asp
Substitution
1A
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
17430
c.99G > C
Substitution
p.Glu33Asp
Substitution
1A
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
17434
c.194A > G
Substitution
p.Glu65Gly
Substitution
1A
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
17435
c.194A > G
Substitution
p.Glu65Gly
Substitution
1A
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
16283
c.302G > C
Substitution
p.Arg101Pro
Substitution
1B
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
17399
c.388G > T
Substitution
p.Ala130Ser
Substitution
1B
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
17755
c.388G > T
Substitution
p.Ala130Ser
Substitution
1B
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
17432
c.471G > A
Substitution
p.=
Silent
Not
LGMD1B
Limb-girdle muscular
159001
#
affected
dystrophy type 1B
9216
c.513G > A
Substitution
p.=
Silent
Not
LGMD1B
Limb-girdle muscular
159001
#
affected
dystrophy type 1B
17575
c.513 + 1G > A
Substitution
p.?
Unknown
Unknown
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
17437
c.565C > T
Substitution
p.Arg189Trp
Substitution
1B
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
9639
c.622_624delAAG
Deletion
p.Lys208del
Deletion
1B
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
12403
c.624_626delGAA
Deletion
p.Lys208del
Deletion
1B
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
16145
c.673C > T
Substitution
p.Arg225X
Substitution
L12
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
17401
c.673C > T
Substitution
p.Arg225X
Substitution
L12
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
17753
c.673C > T
Substitution
p.Arg225X
Substitution
L12
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
13555
c.746G > A
Substitution
p.Arg249Gln
Substitution
2A
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
9164
c.777T > A
Substitution
p.Tyr259X
Substitution
L2
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
12401
c.777T > A
Substitution
p.Tyr259X
Substitution
L2
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
17760
c.777T > A
Substitution
p.Tyr259X
Substitution
L2
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
11810
c.855delG
Deletion
p.Ala287LeufsX191
Frame shift
2B
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
9021
c.864_867delCCAC
Deletion
p.His289ArgfsX190
Frame shift
2B
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
11759
c.864_867delCCAC
Deletion
p.His289ArgfsX190
Frame shift
2B
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
12384
c.908_909delCT
Deletion
p.Ser303CysfsX26
Frame shift
2B
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
11811
c.992G > C
Substitution
p.Arg331Pro
Substitution
2B
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
18309
c.1001_1003delGCC
Deletion
p.Ser334del
Deletion
2B
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
8794
c.1130G > A
Substitution
p.Arg377His
Substitution
2B
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
8850
c.1130G > T
Substitution
p.Arg377Leu
Substitution
2B
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
9301
c.1130G > A
Substitution
p.Arg377His
Substitution
2B
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
9024
c.1130G?T
Substitution
p.Arg377Leu
Substitution
2B
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
11762
c.1130G > A
Substitution
p.Arg377His
Substitution
2B
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
13557
c.1130G > A
Substitution
p.Arg377His
Substitution
2B
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
11755
c.1146C > T
Substitution
p.=
Silent
Not
LGMD1B
Limb-girdle muscular
159001
#
affected
dystrophy type 1B
11756
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
17443
c.1357C > T
Substitution
p.Arg453Trp
Substitution
Tail
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
11812
c.1370delA
Deletion
p.Lys457SerfsX21
Frame shift
Tail
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
17441
c.1380 + 1G > A
Substitution
KO
Unknown
Unknown
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
8839
c.1441T > C
Substitution
p.Tyr481His
Substitution
Tail
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
13376
c.1488 + 5G > A
Substitution
p.?
Unknown
Unknown
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
9181
c.1494G > T
Substitution
p.Trp498Cys
Substitution
Tail
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
9158
c.1494G > T
Substitution
p.Trp498Cys
Substitution
Tail
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
16442
c.1494G > T
Substitution
p.Trp498Cys
Substitution
Tail
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
9159
c.1535T > C
Substitution
p.Leu512Pro
Substitution
Tail
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
11763
c.1535T > C
Substitution
p.Leu512Pro
Substitution
Tail
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
17449
c.1535-T > C
Substitution
p.Leu512Pro
Substitution
Tail
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
17452
c.1535T > C
Substitution
p.Leu512Pro
Substitution
Tail
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
11758
c.1583C > A
Substitution
p.Thr528Lys
Substitution
Tail
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
13446
c.1608 + 1G > A
Substitution
p.?
Unknown
Unknown
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
9412
c.1608 + 5G > C
Substitution
p.Glu537ValfsX36
Frame shift
Tail
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
13151
c.1609-3C?G
Substitution
p.?
Unknown
Unknown
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
11764
c.1718C > T
Substitution
p.Ser573Leu
Substitution
Tail
LGMD1B
Limb-girdle muscular
159001
#
dystrophy type 1B
13505
c.373G > A
Substitution
p.Gly125Ser
Substitution
1B
LAF
Lone atrial fibrillation
—
—
13509
c.[373G > A; =] +
Substitution
p.[Gly125Ser; =] +
Substitution
1B, Tail
LAF
Lone atrial fibrillation
—
—
[=; 1243G > A]
[=; Val415Ile]
13492
c.1310 + 63C > A
Substitution
p.?
Unknown
Unknown
LAF
Lone atrial fibrillation
—
—
13495
c.937 − 46A > G
Substitution
p.?
Unknown
Unknown
LAF
Lone atrial fibrillation
—
—
13501
c.1149G > A
Substitution
p.=
Silent
Not
LAF
Lone atrial fibrillation
—
—
affected
13497
c.1158 − 44C > T
Substitution
p.?
Unknown
Unknown
LAF
Lone atrial fibrillation
—
—
13499
c.1158 − 44C > T
Substitution
p.?
Unknown
Unknown
LAF
Lone atrial fibrillation
—
—
13507
c.1243G > A
Substitution
p.Val415Ile
Substitution
Tail
LAF
Lone atrial fibrillation
—
—
13511
c.1462A > C
Substitution
p.Thr488Pro
Substitution
Tail
LAF
Lone atrial fibrillation
—
—
13503
c.1803C > T
Substitution
p.=
Silent
Not
LAF
Lone atrial fibrillation
—
—
affected
11783
c.176T > G
Substitution
p.Leu59Arg
Substitution
1A
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
18470
c.683A > T
Substitution
p.Glu228Val
Substitution
L12
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
12380
c.[1318G > A; =] +
Substitution
p.[Val440Met; =] +
Substitution
Tail
MADA
Mandibuloacral dysplasia with
248370
#
[=; 1580G > A]
[=; Arg527His]
type A lipodystrophy
12615
c.1411C > T
Substitution
p.Arg471Cys
Substitution
Tail
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
13119
c.1579C > T
Substitution
p.Arg527Cys
Substitution
Tail
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
17878
c.1579C > T
Substitution
p.Arg527Cys
Substitution
Tail
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
8851
c.1580G > A
Substitution
p.Arg527His
Substitution
Tail
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
11419
c.1580G > A
Substitution
p.Arg527His
Substitution
Tail
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
11420
c.1580G > A
Substitution
p.Arg527His
Substitution
Tail
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
11421
c.1580G > A
Substitution
p.Arg527His
Substitution
Tail
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
11422
c.1580G > A
Substitution
p.Arg527His
Substitution
Tail
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
8877
c.1580G > A
Substitution
p.Arg527His
Substitution
Tail
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
11453
c.1580G > A
Substitution
p.Arg527His
Substitution
Tail
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
8995
c.1580G > A
Substitution
p.Arg527His
Substitution
Tail
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
9224
c.1580G > A
Substitution
p.Ala527His
Substitution
Tail
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
17602
c.1580G > T
Substitution
p.Arg527Leu
Substitution
Tail
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
17603
c.1580G > T
Substitution
p.Arg527Leu
Substitution
Tail
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
17747
c.1580G > T
Substitution
p.Arg527Leu
Substitution
Tail
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
17748
c.1580G > T
Substitution
p.Arg527Leu
Substitution
Tail
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
12598
c.1585G > A
Substitution
p.Ala529Thr
Substitution
Tail
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
9317
c.1586C > T
Substitution
p.Ala529Val
Substitution
Tail
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
18478
c.1620G > A
Substitution
p
Substitution
Tail
MADA
Mandibuloacral dysplasia with
248370
#
type A lipodystrophy
13054
c.82C > T
Substitution
p.Arg28Trp
Substitution
Head
—
Metabolic syndrome
—
—
13056
c.274C > T
Substitution
p.Leu92Phe
Substitution
1B
—
Metabolic syndrome
—
—
13059
c.11590 > G
Substitution
p.Leu387Val
Substitution
2B
—
Metabolic syndrome
—
—
13061
c.1184C > T
Substitution
p.Ser395Leu
Substitution
Not
—
Metabolic syndrome
—
—
affected
13063
c.1196G > A
Substitution
p.Arg399His
Substitution
Tail
—
Metabolic syndrome
—
—
13065
c.1262T > C
Substitution
p.Leu421Pro
Substitution
Tail
—
Metabolic syndrome
—
—
13067
c.1315C > T
Substitution
p.Arg439Cys
Substitution
Tail
—
Metabolic syndrome
—
—
13069
c.1516C > G
Substitution
p.His506Asp
Substitution
Tail
—
Metabolic syndrome
—
—
13047
c.1698C > T
Substitution
p.=
Silent
Not
—
Metabolic syndrome
—
—
affected
13071
c.1961_1962insG
Insertion
p.Thr655AsnfsX49
Frame shift
Tail
—
Metabolic syndrome
—
—
12408
c.73C > T
Substitution
p.Arg25Cys
Substitution
Head
—
Muscular dystrophy
—
—
12410
c.1130G > T
Substitution
p.Arg377Leu
Substitution
2B
—
Muscular dystrophy
—
—
12412
c.1622G > C
Substitution
p.Arg541Pro
Substitution
Tail
—
Muscular dystrophy
—
—
12414
c.1045C > T
Substitution
p.Arg349Trp
Substitution
2B
—
Muscular dystrophy and
—
—
lipodystrophy
17411
c.1821G > A
Substitution
p.=
Silent
Not
WRS
Progeroid syndrome,
264090
%
affected
neonatal
18482
c.1940T > G
Substitution
p.Leu647Arg
Substitution
Tail
WRS
Progeroid syndrome,
264090
%
neonatal
17009
c.1303C > T
Substitution
p.Arg435Cys
Substitution
Tail
RD
Restrictive dermopathy
275210
#
17731
c.1303C > T
Substitution
p.Arg435Cys
Substitution
Tail
RD
Restrictive dermopathy
275210
#
17801
c.1303C > T
Substitution
p.Arg435Cys
Substitution
Tail
RD
Restrictive dermopathy
275210
#
9166
c.1824C > T
Substitution
p.[=,
Silent, Deletion
Tail
RD
Restrictive dermopathy
275210
#
Val607_Gln656del]
9208
c.1968 + 1G > A
Substitution
p.Gly567_Gln656del
Deletion
Tail
RD
Restrictive dermopathy
275210
#
17980
c.1057C > T
Substitution
p.Gln353X
Substitution
2B
—
Spinal muscular atrophy with
—
—
cardiac involvment
18424
c.868G > A
Substitution
p.Glu290Lys
Substitution
2B
SCD
Sudden cardiac death
115080
#
17189
c.908_909delCT
Deletion
p.Ser303CysfsX26
Frame shift
2B
SCD
Sudden cardiac death
115080
#
17901
c.1334T > A
Substitution
p.Val445Glu
Substitution
Tail
SCD
Sudden cardiac death
115080
#
9022
c.1804G > A
Substitution
p.Gly602Ser
Substitution
Tail
—
Type A insulin resistance
—
—
syndrome
In some embodiments, the nucleic acid molecule according to any aspect of the invention is for use in treating cardiovascular disease in a subject.
In some embodiments, the disease or the cardiovascular disease Is characterised by the presence of at least one Lmna mutation.
Preferably, the cardiovascular disease is selected from the group consisting of laminopethy, cardiomyopathy, such as dilated cardiomyopathy (DCM), dilated cardiomyopathy 1A, dilated cardiomyopathy with conduction system defects, cardiomyopathy with advanced AV block and arrhythmia, lone atrial fibrillation; muscular dystrophy (often associated with cardiomyopathy), such as cardiomyopathy associated with Emery-Dreifuss muscular dystrophy (autosomal dominant), cardiomyopathy associated with Emery-Dreifuss muscular dystrophy (autosomal recessive), cardiomyopathy associated with Limb-girdle muscular dystrophy type 1B, cardiomyopathy associated with congenital muscular dystrophy; premature aging syndromes (thought to be primarily vascular, but may have cardiac involvement) such as cardiomyopathy associated with Atypical Werner syndrome, cardiomyopathy associated with Hutchinson-Gilford progeria syndrome and the like, as well as diseases presented in bold font in Table 1.
According to a third aspect of the invention there is provided an adeno-associated virus vector (AAV) comprising a cardiac troponin T promoter (cTnT), and the transgene according to any aspect of the invention.
According to a fourth aspect of the invention there is provided a pharmaceutical composition comprising the nucleic acid molecule according to any embodiment of the invention for treating a disease.
In some embodiments the disease is a laminopathy.
In some embodiments the pharmaceutical composition comprising the nucleic acid molecule according to the invention is for use in treating a cardiovascular disease in a subject.
According to a fifth aspect of the invention there is provided a method of treating a disease in a subject, the method comprising administration of a pharmaceutically effective amount of the nucleic acid molecule according to any embodiment of the invention, or the pharmaceutical composition of the invention.
In some embodiments of the method of treating a disease in a subject, the disease is characterised by the presence of at least one Lmna mutation.
In some embodiments of the method of treating a disease in a subject, the method comprises:
(i) testing a sample obtained from a subject suspected of having a disease for the presence or absence of at least one Lmna mutation;
wherein the presence of at least one Lmna mutation indicates that the subject is to be administered the pharmaceutical composition of the invention or the nucleic acid molecule of the invention.
In some embodiments of the method of treating a disease in a subject, the Lmna mutation(s) affect(s) lamin A isoform, or lamin C isoform of the Lmna gene, or both lamin A/C isoforms.
In some embodiments of the method of treating a disease in a subject, the disease is selected from the group consisting of restrictive dermopathy, familial partial lipodystrophy (for example, Dunnigan type), mandibuloacral dysplasia with type A lipodystrophy, metabolic syndrome, Charcot-Marie-Tooth disease type 2, Charcot-Marie-Tooth disease type 2B1 and diseases presented in normal font in Table 1.
In some embodiments of the method of treating a disease in a subject, the disease is a cardiovascular disease, wherein the cardiovascular disease is selected from the group consisting of laminopathy, cardiomyopathy, such as dilated cardiomyopathy (DCM), dilated cardiomyopathy 1A, dilated cardiomyopathy with conduction system defects, cardiomyopathy with advanced AV block and arrhythmia, lone atrial fibrillation; muscular dystrophy (often associated with cardiomyopathy), such as cardiomyopathy associated with Emery-Dreifuss muscular dystrophy (autosomal dominant), cardiomyopathy associated with Emery-Dreifuss muscular dystrophy (autosomal recessive), cardiomyopathy associated with Limb-girdle muscular dystrophy type 1B, cardiomyopathy associated with congenital muscular dystrophy; premature aging syndromes (thought to be primarily vascular, but may have cardiac involvement) such as cardiomyopathy associated with Atypical Werner syndrome, cardiomyopathy associated with Hutchinson-Gilford progeria syndrome; and diseases presented in bold font in Table 1.
In some embodiments of the method of treating a disease in a subject, the subject is a non-human mammal, such as a mouse, or a human.
In some embodiments of the method, the mouse is an N195K mouse (Lmna N195K/N195K), or a Lmna conditional knockout (Lmnaflox/flox).
According to a sixth aspect of the invention there is provided use of the pharmaceutical composition according to the invention or the nucleic acid molecule according to the invention in the manufacture of a medicament for treating a disease caused by one or more Lmna mutations.
In some embodiments the disease is Lmna mutation-related cardiovascular disease.
In some embodiments the disease is selected from the group consisting of restrictive dermopathy, familial partial lipodystrophy (for example, Dunnigan type), mandibuloacral dysplasia with type A lipodystrophy, metabolic syndrome, Charcot-Marie-Tooth disease type 2, Charcot-Marie-Tooth disease type 2B1 and diseases presented in normal font in Table 1.
In some embodiments the cardiovascular disease is selected from the group consisting of laminopathy, cardiomyopathy, such as dilated cardiomyopathy (DCM), dilated cardiomyopathy 1A, dilated cardiomyopathy with conduction system defects, cardiomyopathy with advanced AV block and arrhythmia, lone atrial fibrillation; muscular dystrophy (often associated with cardiomyopathy), such as cardiomyopathy associated with Emery-Dreifuss muscular dystrophy (autosomal dominant), cardiomyopathy associated with Emery-Dreifuss muscular dystrophy (autosomal recessive), cardiomyopathy associated with Limb-girdle muscular dystrophy type 1B, cardiomyopathy associated with congenital muscular dystrophy; premature aging syndromes (thought to be primarily vascular, but may have cardiac involvement) such as cardiomyopathy associated with Atypical Werner syndrome, cardiomyopathy associated with Hutchinson-Gilford progeria syndrome and the like, as well as diseases presented in bold font in Table 1.
According to a seventh aspect of the invention there is provided a method for screening for drug candidates capable of inhibiting or disrupting the LINC complex in a cell.
Accordingly, in some embodiments there is provided a method for screening for drug candidates capable of inhibiting the interaction of the proteins of a LINC complex in a cell, which comprises:
(a) combining the proteins of said LINC complex in the presence of a drug to form a first complex;
(b) combining the proteins in the absence of said drug to form a second complex;
(c) measuring the amount of said first complex and said second complex; and
(d) comparing the amount of said first complex with the amount of said second complex, wherein if the amount of said first complex is less than the amount of said second complex, then the drug is a drug candidate for inhibiting the interaction of the proteins of said UNC complex in a cell.
In some embodiments the drug candidate disrupts the protein-protein interaction between SUN and KASH of the LINC complex. Preferably the drug candidate disrupts the interaction between Sun1 and Nesprin-1 proteins.
In some embodiments said screening is an in vitro screening.
In some embodiments said complex is measured by an ELISA method.
In some embodiments recombinant SUN domain is immobilized on a solid surface and recombinant KASH domain is labelled with an enzyme that can generate a colorimetric or chemiluminescent readout. Compounds that fail to inhibit the SUN-KASH interaction will result in a well in the plate where the recombinant SUN would bind to the enzyme-linked KASH domain. Following wash steps and incubation with colorimetric or chemiluminescent enzyme substrates, the presence of the SUN-KASH interaction can be detected in standard plate readers. If the compound can inhibit SUN-KASH interaction, then following the wash step, the KASH domain would be removed, and there would be reduced or no enzymatic reaction in the well.
In some embodiments if the amount of said first complex is less than the amount of said second complex, then said drug is a drug candidate for inhibiting the interaction of said proteins.
In some embodiments said complex is measured by a fluorescence anisotropy method.
In some embodiments the fluorescence anisotropy method employs recombinant SUN and KASH domains. In some embodiments the KASH domain is fluorescently labelled with a fluorescein moiety and fluorescence anisotropy of the KASH domain interacting with SUN domain may be measured using standard equipment such as a plate reader incorporating a fluorescence spectrometer function.
In some embodiments if the amount of said first complex is less than the amount of said second complex there will be a difference in the fluorescence anisotropy of the fluorescent KASH and said drug is a drug candidate for inhibiting the interaction of said proteins.
Certain terms employed in the specification, examples and appended claims are collected here for convenience.
The terms “amino acid” or “amino acid sequence,” as used herein, refer to an oligopeptide, peptide, polypeptide, or protein sequence, or a fragment of any of these, and to naturally occurring or synthetic molecules. Where “amino acid sequence” is recited herein to refer to an amino acid sequence of a naturally occurring protein molecule, “amino acid sequence” and like terms are not meant to limit the amino acid sequence to the complete native amino acid sequence associated with the recited protein molecule.
As used herein, the term “comprising” or “including” is to be interpreted as specifying the presence of the stated features, integers, steps or components as referred to, but does not preclude the presence or addition of one or more features, integers, steps or components, or groups thereof. However, in context with the present disclosure, the term “comprising” or “including” also includes “consisting of”. The variations of the word “comprising”, such as “comprise” and “comprises”, and “including”, such as “include” and “includes”, have correspondingly varied meanings.
Streptococcus pyogenes
Nat. Protoc
Cell
As used herein, the terms “CRISPR-Cas” and “CRISPR” system are used somewhat interchangeably to refer to a microbial adaptive immune system that uses RNA-guided nucleases to cleave foreign genetic elements. It comprises clustered regularly interspeced short palindromic repeats (CRISPRs), a CRISPR-associated (Cas) endonuclease and a synthetic guide RNA that can be programmed to identify and introduce a double strand break at a specific site within a targeted gene sequence. The palindromic repeats are interspaced by short variable sequences derived from exogenous DNA targets known as protospacers, and together they constitute the CRISPR RNA (crRNA) array. Within the DNA target, each protospacer is always associated with a protospacer adjacent motif (PAM), which can vary depending on the specific CRISPR system. CRISPR-Cas9 is a specific version of the system referring to use of RNA-guided Cas9 nuclease, originally derived from , whereby the target DNA must immediately precede a 5′-NGG PAM. Variations of the CRISPR-Cas9 system are known [Ran F A, at al., 8, 2281-2308 (2013); Ran F A, et al., 154, 1380-1389 (2013)], including CRISPR-Cpf1, and although CRISPR-Cas9 has been used herein in the Examples, it is not intended that the present invention be limited to a particular CRISPR-Cas system.
As used herein, the term “dominant negative” refers to a mutation whose gene product adversely affects the normal, wild-type gene product within the same cell. This usually occurs if the product can still interact with the same elements as the wild-type product, but block some aspect of its function. In one example, the transgene is expressed as a protein, and said protein that is functional as a dimer. A mutation that removes the functional domain, but retains the dimerization domain would cause a dominant negative phenotype, because some fraction of protein dimers would be missing one of the functional domains.
As used herein, the term “normal font” in reference to diseases listed in Table 1 refers to those diseases that are in plain text and not in bold text. The term “bold text” has its ordinary meaning.
As used herein, the term “stabiliser polypeptide” or “stabiliser protein” refers to an inert polypeptide which folds into a discrete domain, thereby ensuring that the remainder of the peptide maintains, for example, the proper topology. In one example, the stabiliser protein ensures that the KASH protein maintains proper topology on the endoplasmic reticulum membrane and the outer nuclear membrane. In another example, the stabiliser polypeptide prevents an attached polypeptide from translocating into, for example, the perinuclear space.
As used herein, the term “operably linked” means that the components to which the term is applied are in a relationship that allows them to carry out their inherent functions under suitable conditions. For example, a control sequence which is “operably linked” to a protein coding sequence is ligated thereto, so that expression of the protein coding sequence is achieved under conditions compatible with the transcriptional activity of the control sequences. By way of an example, a first nucleic acid sequence is operably linked with a second nucleic acid sequence when the first nucleic acid sequence is placed in a functional relationship with the second nucleic acid sequence. For instance, a promoter is operably linked to a coding sequence if the promoter affects the transcription or expression of the coding sequence. Generally, operably linked DNA sequences are contiguous and, where necessary to join two protein-coding regions, in the same reading frame.
As used herein, there term “extension” refers to one or more amino acids that can be found attached to the N- or the C-terminus of a desired peptide.
As used herein, the terms “polypeptide”, “peptide” or “protein” refer to one or more chains of amino acids, wherein each chain comprises amino acids covalently linked by peptide bonds, and wherein said polypeptide or peptide can comprise a plurality of chains non-covalently and/or covalently linked together by peptide bonds, having the sequence of native proteins, that is, proteins produced by naturally-occurring and specifically non-recombinant cells, or genetically-engineered or recombinant cells, and comprise molecules having the amino acid sequence of the native protein, or molecules having deletions from, additions to, and/or substitutions of one or more amino acids of the native sequence. A “polypeptide”, “peptide” or “protein” can comprise one (termed “a monomer”) or a plurality (termed “a multimer”) of amino acid chains.
The term “subject” is herein defined as vertebrate, particularly mammal, more particularly human. For purposes of research, the subject may particularly be at least one animal model, e.g., a mouse, rat and the like. In particular, for treatment or prophylaxis of a laminopthy, such as DCM, the subject may be a human.
The term “treatment”, as used in the context of the invention refers to ameliorating, therapeutic or curative treatment.
Without being bound by theory, the inventors submit that the whole basis of the therapy is that disrupting LINC complex function suppresses Lmna mutation. It is further noted that the target of the claimed methods is the SUN-KASH interaction in the LINC complex. The endogenous protein levels should not be affected.
It is also further noted that the transgene (for example, the dominant negative transgene) will not work if the full length SUN domain protein is inserted between the signal sequence and the KDEL, as it will invert the membrane topology of the protein such that the SUN domain is no longer in the perinuclear space/ER lumen. Only the regions following the transmembrane domain can be used, i.e. the luminal domain.
A person skilled in the art will appreciate that the present invention may be practiced without undue experimentation according to the methods given herein. The methods, techniques and chemicals are as described in the references given or from protocols in standard biotechnology and molecular biology text books.
Flx/Flx
Δ/Δ
Flx/Flx
Flx/Flx/NlMhc
Flx/Flx
Flx/Flx
−/−
N195K/N195K
Δ/Δ
−/−
Flx/
−/−
+/−
−/−
Differentation; research in biological diversity
Cell
Genesis
Genesis
Development
Hum Mol Genet
FIG. 33
FIG. 35
Mice were maintained at the A*STAR Biological Resource Centre facility and the NUS Animal Facility in accordance with the guidelines of the Institutional Animal Care and Use Committee for each facility. The Lmnamice were generated and characterized as previously described [A. S. Wang, at al., , (2015); I. Solovei et al., 152: 584-598 (2013)] (). To derive mice with a global deletion Lmna (Lmna), we crossed the floxed allele (Lmna) to mice in which Cre recombinase is driven by the regulatory sequences of the mouse zona pellucida 3 gene (Zp3;Tg(Zp3-cre)93Knw, JAX stock 003651) [W. N. de Vries et al., 28:110-112 (2000)]. To obtain cardiomyocyte-specific deletion of Lmna (Lmna), we first crossed the Lmnamice to mice in which Cre expression was driven by the cardiac-specific murine alpha myosin-heavy chain (Myh6, myosin, heavy polypeptide 6, cardiac muscle, alpha) promoter (MyHC;Tg(Myhca-cre)2182Mds, JAX stock 011038). To obtain a tamoxifen inducible cardiomyocyte-specific deletion of Lmna (LmnFlx/Flx:mcm), we crossed the Lmnawith mice in which Cre expression was driven by the mouse cardiac-specific alpha-myosin heavy chain promoter (αMHC or alpha-MHC;Myh6) that expressed a tamoxifen-inducible Cre recombinase (MerCreMer) specifically in juvenile and adult cardiac myocytes (mcm;Tg(Myh6-cre/Esr1*)1Jmk, JAX stock 005657). The specificity of mcm Cre expression to cardiomyocytes was confirmed by crossing Cre lines to the mT/mG reporter mice [M. D. Muzumdar, et al., 45: 593-605 (2007)] (). Generation of the Sun1mice was previously described [Y. H. Chi et al., 136: 965-973 (2009)] as were the Lmnamice [L. C. Mounkes, et al., 14: 2167-2180 (2005)]. The Lmna:Sun1 and LmnaFlxmcm:Sun1 mice were obtained by crossing the respective Lamin-Cre mice strains with Sun1 mice as Sun1 mice are infertile.
To test for the insertion of loxP sites and conditional deleted allele, genotyping was performed with a duplex PCR protocol with the following primers were used:
SEQ ID NO: 16
FDK/FLX-F1: 5′-CCAGCTTACAGAGCACCGAGCT-3′,
SEQ ID NO: 17
FLX/FLX-F2: 5′-TCCTTGCAGTCCCTCTTGCATC-3′,
SEQ ID NO: 18
FLX/FLX-R1: 5′-AGGCACCATTGTCACAGGGTC-3′.
To test for Sun1 deletion, the following primers were used:
SEQ ID NO: 19
Sun1-F:
5′-GGC AAG TGG ATC TCT TGT GAA TTC TTG AC-3′
SEQ ID NO: 20
Sun1-R:
5′-GTA GCA CCC ACC TTG GTG AGC TGG TAC-3′
SEQ ID NO: 21
Sun1-E8:
5′-AGC CAC ATA ACC ACC TGG AG-3′
To test for the MyHC transgene, the following primers were used:
SEQ ID NO: 22
MyHC-tF: 5′-ATG ACA GAC AGA TCC CTC CTA TCT CC-3′
SEQ ID NO: 23
MyHC-tR: 5′-CTC ATC ACT CGT TGC ATC ATC GAC-3′
SEQ ID NO: 24
MyHC-F: 5′-CAA ATG TTG CTT GTC TGG TG-3′
SEQ ID NO: 25
MyHC-R: 5′-GTC AGT CGA GTG CAC AGT TT-3′
To test for the presence of mcm transgene, the following primers were used:
SEQ ID NO: 26
mcm-3798t: 5′-AGG TGG ACC TGA TCA TGG AG-3′
SEQ ID NO: 27
mcm-8346t: 5′-ATA CCG GAG ATC ATG CAA GC-3′
SEQ ID NO: 28
mcm-7338: 5′-CTA GGC CAC AGA ATT GAA AGA TCT-3′
SEQ ID NO: 29
mcm-7339: 5′-GTA GGT GGA AAT TCT AGC ATC ATC C-3′
2
2
2
Young mice (14 days old) and adult mice (3-5 months old) were injected once with 40 mg/kg of Tamoxifen (Sigma) dissolved in Corn Oil (Sigma). Mice were sacrificed by COeuthanasia or anesthetised with a gaseous mixture of 1.5% Isoflurane (BioMac) and 1.5LOat various time points after tamoxifen injection. Cardiac arrest was induced by injection of 15% KCl, followed by flushing with PBS to remove blood. Hearts for paraffin embedding were additionally flushed with 4% paraformaidehyde (PFA), left in 4% paraformaldehyde (PFA) overnight, dehydrated in 70% ethanol for at least 24 hr and embedded in paraffin. Hearts for cryosection were embedded in tragacanth gum (Sigma), frozen in isopentane (BDH-AnalaR) cooled in liquid N, cut 9 μm sections by cryostat (Leica CM3050), collected onto charged slides and stored at −20° C. for histological and immunofluorescence staining. Hearts for protein and RNA extraction were snap frozen in liquid N2 and stored for further processing.
Circulation Research
2
2
Cardiomyocyte isolation was carried out as per standard protocol [M. Ackers-Johnson et al., 119: 909 (2016)]. Briefly, mice were anaesthetised with isoflurane (100% Oat 0.5 L/min, isoflurane atomiser dial at 4%). Mice hearts were stopped with 15% KCl, descending aorta was cut and hearts were flushed with 7 mL of EDTA buffer into the right ventricle. Ascending aorta was clamped using Reynolds forceps, the entire heart removed and placed in a 60 mm dish containing fresh EDTA buffer. Hearts were digested by sequential injections of 10 mL EDTA buffer, 3 mL Perfusion buffer and 30-50 mL Collagenase buffer into the left ventricle. Forceps were used to gently pull the digested heart into smaller pieces ˜1 mm and gentle trituration. Enzymatic activity was inhibited by addition of 5 ml of Stop buffer. Cell suspension was passed through a 100 um filter, and four sequential rounds of gravity settling to enrich for myocytes, ultimately obtaining a highly pure myocyte fraction. The myocyte pellet was snap frozen in liquid Nand stored a −80° C. until further processing.
For histological studies, sections (9 μm) were stained with standard Hematoxylin and Eosin for cell morphology, Masson's trichrome stain to detect collagen and TUNEL assay to detect apoptotic nuclei. Images were obtained on a Zeiss Axio Imager Microscope. For immunofluorescence on frozen heart sections, sections were warmed to room temperature, rehydrated with PBS, blocked with M.O.M block (Vector Shields) and donkey serum (Sigma-Aldrich), incubated with primary antibodies overnight at 4° C. The slides were then washed in PBS and incubated with secondary antibodies and Hoechst dye (Sigma-Aldrich) for 60 mins, washed with PBS and mounted in Prolong-Gold Anti-fade reagent (Invitrogen). Primary antibodies: LMNA/C N-18 (goat, 1:50, Santa Cruz), Sun1 monoclonal (mouse, neat, from B. Burke), PCM-1 (rabbit, 1:200, Sigma) and sarcomere-α-actinin (mouse, 1:100, abcam); Secondary antibodies were: Alexa Fluor 488, 568 and 847 (1:250, Invitrogen). For isolated cardiomyocyte immunofluorescence, myocytes were stained in suspension and spun down gently for each solution change then plated on glass slides for imaging with a Zeiss LSM510 inverted confocal microscope.
Whole Hearts and Quadriceps muscles were homogenized in RIPA lysis buffer and spun at 13,200 g, 10 min, 4° C. Total cell lysates were electrophoresed and transferred to PVDF membrane and blocked with Odyssey Blocking Buffer (Li-Cor Biosciences). The membrane was incubated with primary antibodies for 2 h at room temperature. After which, membrane was washed in TBST washing solution and incubated in Odyssey IRDye secondary antibodies for 1 h before visualization on the Odyssey Infrared Imaging System (Li-Cor Biosciences). The primary antibodies used for detection of LMNA/C (Rabbit, Cell Signalling) that is specific to an epitope in the first 50 amino acids in LMNA, Sun1 monoclonal (mouse, 1:500, Burke) and control beta-tubulin (rabbit, 1:1000, Abcam).
J Physiol
Mouse papillary muscle from mouse left ventricle was prepared according to the methods described before [C. N. Toepfer, et al., 594: 5237-5254 (2016)]. Briefly, explanted mouse heart was immediately rinsed with oxygenated ice-cold Krebs-Henseleit solution with 12 unit/mL heparin sodium (EDQM) and 30 mM 2,3-Butanedione monoxime (BDM, Sigma) and excess blood was removed. After that, the heart was transferred to ice-cold Krebs-Henseleit solution in a glass petri-dish under a dissection microscope with a cooling stage. Cylindrical papillary (200-300 μm in diameter and 1.5-2 mm in length) were excised from the left ventricle. T-shaped aluminium clips with a hole were crimped onto the ends of a papillary preparation and the prepared papillary chunks were fixed using pins onto a glass petri-dish with a layer of PDMS sylgard 184 (Dow Corning). Papillary preparations were immersed in a 2% Triton X-100 solution at 4° C. overnight.
J Biol Chem
Force measurement was performed as previously described [C. Toepfer et al., 288: 13448-13454 (2013)]. The T-shaped aluminium clips at the ends of the papillary preparations were attached to the hooks of a force transducer (AE801, HJK Sensoren+Systeme) and servo-motor in the experimental rig and were glued with shellac in ethanol (Sigma) to minimize the movement during the experiment. Papillary contraction force was measured at 20° C. The max contraction force was measured in activing solution (100 mM TES, 6.5 Mm MgCl2, 25 mM Ca-EGTA, 5.7 mM Na2ATP, 20 mM Glutathione, 21.5 mM sodium creatine phosphate, pH=7.1, Ionic strength is 150 mmol/L) with 32 μmol/L free Ca2+. The data were collected and processed from the force transducer and DAQ data acquisition device (National Instrument) using a customized software programmed by LabVIEW 2013 (National instrument). At least 5 fibres were tested in each mouse, and at least 3 mice were tested for each experimental group.
J Cell Biol.
2
2
The DN-Sun1 (SS-HA-Sun1L-KDEL) and GFP (SS-GFP-KDEL) vectors were as described [M. Crisp et al., 172: 41-53 (200)]. Briefly, almost the entire lumenal domain of Sun1 was tagged at its NHterminus with HA (HA-Sun1L). To introduce the HA-Sun1L as a soluble form into the lumen of the ER and PNS, signal sequence and signal peptidase cleavage site of human serum albumin was fused onto the NHterminus of HA-Sun1L to yield SS-HA-Sun1L To prevent its secretion, a KDEL tetrapeptide was fused to the COOH terminus of SS-HA-Sun1L to form the final SS-HA-Sun1L-KDEL. The HA-Sun1L region was replaced with GFP sequence to generate the SS-GFP-KDEL.
FIG. 10
The DN-Sun1 and GFP fragments were amplified with the primers listed below (same forward primer was used for both fragments) and ligated into pENN-AAV-cTnT-Pl-eGFP plasmid (kind gift from Dr J. Jian), digested with NcoI and KpnI, to produce Penn-AAV-cTnT-Sun1DN (; SEQ ID NO: 3).
SEQ ID NO: 30
aav Sun1 F
5′-CgagaattcacgcgggccgccATGAAGTGGGTAACCTTTATTTC-3′
SEQ ID NO: 31
aav Sun1 R
5′-CgggtcgactctagaggtaccttaCTACAACTCATCTTTCTGGATG-
3′
SEQ ID NO: 32
aav GFP Sun R
5′-CgggtcgactctagaggtacttaCTACAACTCATCTTIGGATCC-3′
All restriction enzymes were purchased from NEB. PCR reactions were conducted using Q54 Hot Start High-Fidelity 2× Master Mix (NEB, M0494L). Ligations were conducted using isothermal assembly with NEBuilder® HiFi DNA Assembly Master Mix (NEB, E2621L). Primers used for constructing the plasmids were ordered from IDT.
Current Protocols in Molecular Biology
AAV Viruses were produced as per standard protocol [H. Wakimoto, et al., in . (John Wiley & Sons, Inc., 2001)]. Materials supplied by R. Foo: pAAV2/9- the trans-plasmid encoding AAV replicase and capsid gene (SEQ ID NO: 2; available from University of Pennsylvania Penn Vector Core); pAdDeltaF6—the adenoviral helper plasmid (SEQ ID NO: 1) (available from University of Pennsylvania Penn Vector Core); QIAGEN Plasmid Maxi Kit; HEK293T cells (ATCC); Transfection reagent (polyethylenimine e.g., Polysciences). AAV-DJ capsid was obtained from Cell Biolabs, Inc. The pAAV2/9, AAV-DJ, pAdDeltaF6, DN-Sun1 and GFP plasmids were purified using a QIAGEN Plasmid Maxi Kit HEK293T cells were transfected with the virus combination of pAAV2/9, pAdDeltaF6 and either DN-Sun1 or GFP plasmids. Cells were collected and virus purified by lodixnol gradient ultracentrifugation.
2
The following timeline was used for infection of the mouse hearts. Mice were genotyped at 10 days postnatally. They were then subjected to 1 IP injection of Tmx (40 mg/kg of mouse weight) at 14 days postnatally, followed by a concentration of 5×10∧10 vg/g AAV9-DN-Sun1 or AAV9-GFP virus injected into the thoracic cavity at 15 days postnatally. Adult mice (3-5 months old) were injected IP with a single dose of Tmx (40 mg/kg of mouse weight), followed by injection of AAV at a concentration of 5×10∧10 vg/g AAV9-DN-Sun1 or AAV9-GFP virus into the thoracic cavity. Young and adult mice were anesthetised with a gaseous mixture of 1.5% Isoflurane (BioMac) and 1.5 L Obefore virus injections.
Plasmid Construction and Generation of Cas9 mRNA and sgRNAs
pX330 was obtained from Addgene (#42230, Cambridge, Mass., USA). The 20 nt Sun1 and Syne1 single guide RNA (sgRNA) sequences were designed with the help of CRISPR Design Tool (crispr.genome-engineerng.org). A region of the gene of interest was submitted to the tool to identify suitable target sites. Since off-target mutations are possible in CRISPR/Cas9-mediated targeted mutagenesis in the mouse, the CRISPR Design Tool is able to experimentally assess off-target genomic modifications for each gRNA target sites and provide computationally predicted off-target sites for each intended target, ranking the target sequence according to quantitative specificity analysis on the effects of base-pairing mismatch identity, position and distribution. Complimentary oligonucleotides containing the gRNA target sequences were annealed and cloned into the BbsI site of pX330. Guide RNA sequences were as follows:
(SEQ ID NO: 33)
5′-GCACAATAGCCTCGGATGTCG-3′ for Sun1ΔSUN,
(SEQ ID NO: 34)
5′-CCGTTGGTATATCTGAGCAT-3′ for Syne1-stop,
(SEQ ID NO: 35)
5′-GGTTATGGCCGATAGGTGCAT-3′ for Tyrosinase4a
These plasmids (pSun1ΔSUN, pSyne1-stop and pTyrosinase4a) were then sequenced to verify correct insertion of the target sequences. For in vitro transcription, PCR was performed to generate the appropriate transcription templates using a common reverse primer (AAAAGCACCGACTCGGTGCC-3′; SEQ ID NO: 36) and gRNA-specific forward primers that encoded the T7 promoter sequence as follows:
(SEQ ID NO: 37)
Sun1ΔSUN:
5′-TTAATACGACTCACTATAGCACAATAGCCTCGGATGTCG-3′;
(SEQ ID NO: 38)
Syne1-stop:
5′-TTAATACGACTCACTATAGCCGTTGGTATATCTGAGCAT-3′;
(SEQ ID NO: 39)
Tyrosinase4a:
5′-TTAATACGACTCACTATAGGTTATGGCCGATAGGTGCAT-3′
The gRNA PCR products were then subjected to agarose gel electrophoresis (1.5% agarose) to confirm successful PCR, gel purified and used as templates for in vitro transcription using the MEGAshortscript T7 kit (Life Technologies). The gRNAs were purified using MEGAclear kit (Life Technologies) and eluted in RNase-free water. A sample of purified gRNAs were then subjected to agarose gel electrophoresis for quality checks before injecting into zygotes.
2
2
3 to 4 weeks old C57BL/6N females were superovulated with Pregnant Mare Serum gonadotropin (Calbiochem, 38722, 5 IU/ml). 48 hours later, the females were injected with human chorionic gonadotropin (Sigma, CG10, 5 IU/ml) and were mated with C57BL6 males. The following day, fertilized 0.5 dpc embryos were collected from the oviducts. Cas9 mRNA (Sigma, CAS9MRNA, 100 ng/ul), Tyrosinase4a gRNA (50 ng/ul) and gene-specific gRNA (50 ng/ul) were co-injected into the cytoplasm of the embryos in M2 medium (EmbryoMax® Sigma) using a microinjection system (Nikon). Syne1-stop sgRNA were used to derive Syne1 C′T mutant mice and Sun1ΔSUN sgRNA were used to derive Sun1ΔSUN mutant mice. The injected zygotes were cultured in KSOM with amino acids (EmbryoMax® Sigma) in an incubator maintained at 37° C. with 5% COand 5% Ofor 2 hours before implanting into 0.5 dpc pseudopregnant C3H-ICR females.
Mouse tails were clipped and each placed in a 1.5 ml Eppendorf tube. 80 μl of lysis buffer (25 mM NaOH, 0.2 mM EDTA, pH 12) was dispensed into the tube and heated at 95° C. for 60 minutes. After heating, the buffer was neutralized with an equal volume of 40 mM Tris-HCl, pH 5. For certain applications, DNA was extracted and purified from mouse tails using DNeasy Blood and Tissue Kit (QIAGEN).
CRISPR modified mutant mice were genotyped by PCR followed by gel electrophoresis using a high resolution agarose (2% MetaPhor agarose, Lonza).
Primers for Syne1CT′Δ8 Mice were:
SEQ ID NO: 40
Forward: 5′-TGCTCCTGCTGCTGCTTATT-3′
and
SEQ ID NO: 41
Reverse: 5′- ACATGGTGGAGCATTTGTCTCC -3′
Primers for Sun1 CRISPR Mice were:
SEQ ID NO: 42
Forward: 5′-TGACCTTGAGCTGAAACTGC-3′
and
SEQ ID NO: 43
Reverse: 5′-TCAGAACACTGGCACACACA-3′
Lmna mutant mice were genotyped as described in Example 1. To determine sequence of CRISPR-Induced mutations, PCR products from mouse tall DNA were subjected to TOPO cloning (Zero Blunt™ TOPO™ PCR Cloning Kit, 450245, Thermo Fisher Scientific). Plasmid DNA from at least 10 bacterial colonies were isolated using a mini-prep kit (QIAGEN, QIAprepSpin, Miniprep Kit) and sent for Sanger sequencing.
To isolate myoblasts, limbs were obtained from euthanized mice and muscles were dissected from bone. Tissue digestion was performed by incubating the muscle tissues in enzyme solution consisting of equal volumes of dispase II (Roche, cat. 04942078001) at a concentration of 2.4 U/ml and 1% collagenase II (GIBCO® Invitrogen, cat 17101-015) in a 37° C. water bath for 30 minutes, with occasional mixing at 10 minutes interval. After 30 minutes, enzyme solution was neutralized in D10 media (Dulbeco's Modified Eagle Medium (DMEM) with 10% fetal bovine serum). Mixture is then filtered through 70 μm sterile filter (BD Falcon™, cat 352350) and 40 μm sterile filter (BD Falcon™, cat 352340). The suspension was then centrifuged, supernatant removed and subsequently resuspended in F10 media (GIBCO® Invitrogen, cat. 11550043) supplemented with 10 μg/ml bFGF (GIBCO®, cat PHG0264) and plated in 100 mm plates. Mouse adult fibroblasts were allowed to settle for 2 to 3 hours before collecting the supernatant (with floating myoblasts) and replated into 60 mm plates coated with 0.15% Gelatin (Sigma, cat G1393). D10 media was added to the 100 mm plates with MAFs. To terminally differentiate myoblasts to myotubes, the media was changed to DMEM supplemented with 2% horse serum (Thermo Fisher Scientific GIBCO®, cat 16050122).
Whole cell lysates were generated using the Lysis-M kit solution (cOmplete; Roche). Cells were washed in ice-cold PBS and lysed with Roche Lysis M buffer, and centrifuged at 14,000 g for 10 minutes to remove cell debris. To extract protein from tissue sample, small slices of tissue were rapidly placed into Lysing Matrix D tubes (MP Biomedicals), and snap frozen in liquid nitrogen. After snap freezing, the tubes were either stored at −80° C. or used directly for protein analysis. Protein extraction buffer (50 mM Tris (pH7.4), 500 mM NaCl, 0.4% SDS, 5 mM EDTA (pH7.4), 1× Protease inhibitor (cOmplete™ EDTA-free Protease Inhibitor cocktail, Cat no. 04693159001, Roche), 2% Triton, 1 mM Dithiothreitol, in distilled water) was added to tissues, which were then homogenized using the FastPrep™-24 Instrument (MP Biomedicals). Samples were then centrifuged at 14,000 g for 10 minutes to remove cell debris. Protein concentration was quantified using bicinchoninic acid (BCA) protein kit (Bio-Rad) before loading protein samples onto a polyacrylamide gel to ensure equal amounts were being analyzed. All protein samples were resolved by SDS-PAGE gel analysis and transferred onto polyvinylidene fluoride (PVDF) membrane (Millipore) by wet transfer for 48 hours at 20V at 4° C. Membranes were blocked in TBS containing 0.1% Tween 20 (TBST) supplemented 5% milk powder (Anlene) for 1 hour at room temperature. Western Blot analysis was performed using primary antibodies diluted in 5% milk powder (diluted in TBST). Membranes were incubated for 2 hours at room temperature or overnight at 4° C. For secondary antibodies, horseradish-peroxidase (HRP) (Invitrogen) conjugated antibodies were used for chemiluminescent imaging. The membranes were incubated for 1 hour at room temperature with the secondary antibodies. For immunoblots visualized by chemiluminescence, membranes were incubated in ECL substrate (Pierce) for 1 minute before being exposed to a chemiluminescence sensitive film (Thermo Scientific) and subsequently processed.
Cells were grown in 8-well slides (Ibidi) and fixed in ice-cold methanol for 15 minutes at −20° C. They were then rinsed in PBS twice and permeabilized and blocked with 0.1% Triton X, 3% BSA in PBS for 15 minutes at room temperature. The fixed and permeabilized cells were then rinsed in PBS three times. Samples were then incubated with primary antibodies (Table 2) for 2 hours at room temperature or overnight at 4° C. Samples were then washed with PBS three times and subsequently incubated with secondary antibodies (Life Technologies) and DAPI (Life Technologies) for 1 hour at room temperature. After three washes in PBS, cells were mounted in Anti-fade (1% DABCO, 90% Glycerol, 10% PBS) and inspected using a Zeiss 510 Meta Confocal microscope or Axiovert 200 inverted epifluorescence microscope (Zeiss). Images were recorded and analysed using Zeiss ZEN, Metamorph or Image J (NIH) software.
TABLE 2
Antibodies used for immunofluorescence study.
Antibody
Type and Source
Concentration
Akap450, HPA-026109
Polyclonal, Sigma
1:500
MF20
Monoclonal, DSHB
1:25
Nesp1 (MANNES1A)
Monoclonal, Glenn Morris
1:1000 (Western)
1:50 (IF)
Nesp1-C′T
Monoclonal, Brian Burke
Undiluted
supernatant
LaminA, ab8984
Monoclonal, Abcam
1:200
LaminA, SSD
Monoclonal, Brian Burke
1:200
Pcm-1, HPA-023374
Polyclonal, Sigma
1:100
Pcnt, ab4448
Polyclonal, Abcam
1:100
Sun1-9F10
Monoclonal, Brian Burke
1:200
Sun2-3.1E
Monoclonal, Brian Burke
1:500
Nesprin-2
Polyclonal, MyBiosource.com
1:500
Δ/Δ
C′TΔ8/C′TΔ8
Flx/Flxmcm
C′TΔ8/C′TΔ8
Δ/+
Flx/Flx:mcm
C′TΔ8/C′TΔ8
C′TΔ8/+
C′TΔ8/C′TΔ8
+/−
−/−
Lmna mice and tamoxifen injection were described in Example 1. To obtain Lmna:Syne1and Lmna:Syne1double mutant mice, Lmna or Lmnamice were intercrossed with Syne1mice. In the Syne2 mouse model, a IRES-β-gal neomycin selectable cassette (PgkNeo) flanked by IoxP sites was inserted into the Syne2 gene, resulting in deletion of part of axon 102 and all of exons 103-104. The neomycin cassette was subsequently removed by crossing with Cre recombinase mice. Syne1 or Syne1mice were crossed with Syne2 or Syne2 mice to obtain mice with mutant Syne1 and Syne2 alleles, which were intercrossed to obtain double mutant mice. Kaplan-Meier method was used to draw the survival curves.
Potential guide RNA sequences to disrupt human SYNE1 KASH domain or SUN1 SUN domain were determined using CRISPR tool in Benchling software (Benchling Inc. USA) and are shown in Table 3.
TABLE 3
Potential guide RNA sequences to target final exons in human SYNE1 (Nesprin-1) or SUN1 genes
SEQ
ID
Gene Name
ENSEMBL gene ID
Chromosome
Position
Strand
Sequence
PAM
CRISPR enzyme
NO:
SYNE1
ENSG00000131018
6
515330
−
TCGTGTATCTGAGCATGGGG
TGGAAT
saCas9
44
SYNE1
ENSG00000131018
6
515335
−
GCCATTCGTGTATCTGAGCA
TGGGGT
saCas9
45
SYNE1
ENSG00000131018
6
515340
+
TCCACCCCATGCTCAGATAC
ACGAAT
saCas9
46
SYNE1
ENSG00000131018
6
515320
−
GAGCATGGGGTGGAATGACC
GGG
spCas9
47
SYNE1
ENSG00000131018
6
515321
−
TGAGCATGGGGTGGAATGAC
CGG
spCas9
48
SYNE1
ENSG00000131018
6
515330
−
TCGTGTATCTGAGCATGGGG
TGG
spCas9
49
SYNE1
ENSG00000131018
6
515333
−
CATTCGTGTATCTGAGCATG
GGG
spCas9
50
SYNE1
ENSG00000131018
6
515334
−
CCATTCGTGTATCTGAGCAT
GGG
spCas9
51
SYNE1
ENSG00000131018
6
515335
−
GCCATTCGTGTATCTGAGCA
TGG
spCas9
52
SYNE1
ENSG00000131018
6
515345
+
CCCATGCTCAGATACACGAA
TGG
spCas9
53
SYNE1
ENSG00000131018
6
515333
+
CCCGGTCATTCCACCCCATG
TTTG
Cpf1
54
SUN1
ENSG00000164828
7
873276
+
TTTTTCTAACTGGGGCCATC
CTGAGT
saCas9
55
SUN1
ENSG00000164828
7
873285
−
CCGATACAGACAGGTATACT
CAGGAT
saCas9
56
SUN1
ENSG00000164828
7
873266
+
AACTTCGGATTTTTTCTAAC
TGG
spCas9
57
SUN1
ENSG00000164828
7
873267
+
ACTTCGGATTTTTTCTAACT
GGG
spCas9
58
SUN1
ENSG00000164828
7
873268
+
CTTCGGATTTTTTCTAACTG
GGG
spCas9
59
SUN1
ENSG00000164828
7
873280
−
ACAGACAGGTATACTCAGGA
TGG
spCas9
60
SUN1
EN5G00000164828
7
873296
+
CTGAGTATACCTGTCTGTAT
CGG
spCas9
61
SUN1
ENSG00000164828
7
873281
+
TTCTAACTGGGGCCATCCTG
TTTT
Cpf1
62
SUN1
ENSG00000164828
7
873282
+
TCTAACTGGGGCCATCCTGA
TTTT
Cpf1
63
SUN1
ENSG00000164828
7
873283
+
CTAACTGGGGCCATCCTGAG
TTTT
Cpf1
64
SUN1
ENSG00000164828
7
873284
+
TAACTGGGGCCATCCTGAGT
TTTC
Cpf1
65
PAM, protospacer adjacent motif; saCas9, <i>Staphylococcus aureus</i> Cas9; spCas9, <i>Streptococcus pyogenes</i> Cas9; Cpf1, CRISPR from <i>Prevotella</i> and <i>Francisella</i> 1.
All statistical analysis was performed using Graphpad Prism software.
Flx/Flx
Flx/Flx
Flx/Flx
Δ/Δ
−/−
Δ/Δ
Δ/Δ
Flx/Flx
Δ/Δ
−/−
Flx/Flx
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
+/+/mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
FIG. 33
FIG. 26A
FIG. 28A
FIG. 36
FIG. 26C
FIG. 26C
FIG. 27A
FIG. 27B
FIG. 27C
FIG. 27D
FIG. 27E
FIG. 28A
FIG. 288
FIG. 28B
FIG. 28C
FIG. 28C
FIG. 28D
FIG. 28D
FIG. 28D
Differentiation; research in biological diversity
Cell
Genesis
J Clin Invest
Circ Res
Nature communications
Nucleus
To further define the interaction between Sun1 and Lmna in postnatal pathology in mice, we specifically ablated the Lmna gene in different tissues by using a conditional Lmnaline of mice (), that when recombined by Cre activation, results in the complete loss of LaminA/C protein [A. S. Wang, et al., , (2015); I. Solovei et al., 152: 584-598 (2013)]. When Lmnawas constitutively deleted in all tissues by crossing the Lmnamice with Zp3-Cre mice [W. N. de Vries et al., 26: 110-112 (2000)], the mean postnatal lifespan was 17.5 days (). When the same deletion was induced in the absence of Sun1, the Lmna:Sun1 mice lived to a mean of 32.5 days, almost a doubling in longevity (). Performing the same Lmna deletion on a Sun2 null background did not extend the longevity of Lmnamice, revealing the longevity extension is specific to the loss of Sun1 (). Since the A-type lamins are widely expressed in almost all adult tissues, we then determined to what extent, Lmna deletion, specifically in cardiomyocytes, contributes to the early postnatal death of Lmn mice. Furthermore, the inventors wished to ascertain whether loss of Sun1 would increase longevity in these mice harbouring Lmna deficient cardiomyocytes. We first crossed the Lmnawith a constitutive myh6 Cre [R. Agah et al., 100: 169-179 (1997)], in which Cre expression, though constitutive, is restricted to cardiomyocytes but commences during embryogenesis. These mice survived slightly longer than the Lmna to an average of 26.5 days postnatally (). When the same cardiomyocyte specific deletion was performed on a Sun1 background, this resulted in a significant increase in longevity to at least 6 months and beyond after birth (). To further define the loss of Lmna and its effect in postnatal/adult cardiomyocytes we derived mice homozygous for the Lmnallele carrying the inducible cardiomyocyte specific Cre Tg(Myh6-cre/Esr1), (here abbreviated to mcm) in which Cre is induced by a single injection of tamoxifen (Tmx) [D. S. Sohal et al., 89: 20-25 (2001)]. From this cross, the average lifespan of 3-5 month old Lmnamice, following Cre induction was 27 days (). Controls were unaffected by Tmx injection. PCR and immunofluorescence analysis confirmed the Lmna deletion was specific to the Lmnacardiomyocytes, with no detectable recombination occurring in the brain, diaphragm, lung, liver and skeletal muscle, or in wild-type control animals (). By 21 days post injection, Lmnamice showed laboured breathing, a dishevelled, ungroomed appearance, increased lethargy and kyphosis (). Immunofluorescence analysis of isolated cardiomyocytes (CM) and sections of Lmnahearts showed reduced levels of LaminA protein and cardiomyocyte nuclei without any LaminA expression (). LaminA protein levels were decreased 3.5 fold in Lmnahearts after Cre induction compared to uninduced Lmnaand Lmnahearts (). By sampling Lmnamice at specific time points after Tmx injection, it was estimated that it takes 7-14 days after Cre induction for LMNA protein levels to fall by 50% (data not shown), a rate consistent with a study using siRNA LMNA knockdown in human fibroblasts by 1.3-fold after 48 hrs and a further 4-fold reduction after 10.5 days [A. Buchwalter and M. W. Hetzer, 8: 328 (2017); T. Sieprath et al., 6: 236-248 (2015)]. Echocardiograms (ECGs) performed at 21 days after Cre induction revealed poor cardiac contractility in Lmnamice compared to Lmnacontrols (). There was a significant reduction in the Ejection Fraction (EF %) and Fractional shortening (FS %) (P<0.0001) (). The left systolic and diastolic ventricular internal diameters (LVID) were enlarged, compared to Lmnacontrols (). Significantly fewer viable (brick-like) cardiomyocytes were isolated from Lmna+Tmx hearts compared to Lmnacontrols (). Visual analysis revealed the isolated cardiomyocytes from Lmna+Tmx hearts contained large intracellular vacuoles (). Histological analysis of Lmna+Tmx hearts, revealed infiltration of nucleated cells and increased intercellular spaces between cardiomyocytes compared to Lmnacontrol hearts (). The left ventricular lumen in Lmna+Tmx hearts was significantly enlarged, together with significantly increased levels (P=0.0098) of fibrosis were noted in Lmna+Tmx hearts compared to in Lmnacontrols (). Increased numbers of apoptotic cells were also identified in Lmna+Tmx hearts compared to control hearts (). However there was no evidence of extensive DNA damage detectable in the cardiomyocytes, as assessed by Rad51, MRE11, H2AX phosphor-Ser and 53BP1 immunostaining (Data not shown).
Cell
Flx/Flx:mcm
Flx/Flx:mcm
−/−
Flx/Flx:mcm
+/+
Flx/Flx:mcm
+/+
Flx/Flx:mcm
−/−
Flx/Flx:mcm
−/−
Flx/Flx:mcm
+/+
Flx/Flx:mcm
+/+
Flx/Flx:mcm
−/−
Flx/Flx:mcm
+/+
Flx/Flx:mcm
−/−
FIG. 26C
FIG. 26C
FIG. 29A
FIG. 29A
FIG. 29A
FIGS. 29A
FIG. 29C
FIG. 29C
Mice with Lmna mutations show a significant increase in longevity and health in the absence of Sun1 [C. Y. Chen et al., 149: 565-577 (2012)]. As described, induced deletion of Lmna in cardiomyocytes (Lmna+Tmx) results in death within 1 month post Cre induction (). Strikingly, when the same deletion was induced on a Sun1 null background the mice survived for more than 1 year after Cre induction (). Hearts from LmnaSun1+Tmx mice, 3 weeks after induction, were compared to those from LmnaSun1+Tmx to determine the extent to which SUN1 loss ameliorated the pathological changes induced by Lmna loss in cardiomyocytes. Immunofluorescent imaging for Lamin A/C identified many elongated and distorted nuclei. In some of these, residual Lamin A/C was displaced to one pole of the nucleus ( panel 1 and insert) in the LmnaSun1+Tmx hearts. In contrast in the LmnaSun1+Tmx hearts, while there were many elongated nuclei, these showed few if any distortions, even when there was no Lamin A/C staining ( panel 3 yellow arrow heads). Western analysis of whole hearts revealed a significant reduction in Lamin A/C in the LmnaSun1+Tmx hearts (P=0.0359) lysates compared to LmnaSun1 controls ( lower panels). The LmnaSun1+Tmx cardiomyocyte nuclei exhibited increased longitudinal length, together with a segmented appearance, with the segments connected by narrow bridges ( and C, panel 1 arrows). However, in the absence of Sun1, LmnaSun1 cardiomyocyte nuclei exhibited no abnormalities or segmentation ( panels 3 and 4). In total, 70% of cardiomyocytes in LmnaSun1 mice had ruptured or misshapen nuclei compared to fewer than 1% of the cardiomyocytes from the LmnaSun1 ( panel 5).
Flx/Flx:mcm
+/+
Flx/Flx:mcm
−/−
Flx/Flx:mcm
+/+
Flx/Flx:mcm
−/−
FIG. 29B
FIG. 29B
Clear enlargement of the left ventricle (LV) was evident in the LmnaSun1 mice but not in the LVs of the LmnaSun1+Tmx hearts (, panels 1 and 2). The LmnaSun1 hearts exhibited significantly increased levels of fibrosis (P<0.0001) compared to controls, whereas there was no significant fibrosis in the LmnaSun1 hearts (, panels 3-5).
Flx/Flx:mcm
+/+
Flx/Flx:mcm
+/+
Flx/Flx:mcm
−/−
FIG. 29B
As a model for ventricular muscle mechanics we measured the active force in cardiac papillary muscle. The active force was significantly reduced by 66% in Lmna:Sun1+Tmx papillary muscle (P=0.0028) compared to LmnaSun1+CTL. In the absence of SUN1, LmnaSun1+Tmx cardiac papillary active force was maintained at levels not significantly different from those of controls ( panel 6).
Flx/Flx:mcm
+/+
Flx/Flx:mcm
−/−
Flx/Flx:mcm
−/−
Flx/Flx:mcm
+/+
FIG. 29D
Echocardiograms performed before and after Cre induction revealed progressive worsening of cardiac contractility in the LmnaSun1+Tmx mice compared to LmnaSun1+Tmx mice (). Loss of SUN1 preserved both EF, FS and Global Longitudinal Strain (GLS) (GLS is a separate parameter used to assess myocardial contractility, and Is a better predictor of heart failure) In LmnaSun1+Tmx mice compared to LmnaSun1+Tmx mice.
Flx/Flx:mcm
−/−
Flx/Flx:mcm
−/−
Flx/Flx:mcm
−/−
Flx/Flx:mcm
+/+
Flx/Flxmcm
−/−
Flx/Flx
FIG. 37C
FIG. 37D
FIGS. 37A
FIG. 37E
FIG. 26C
PCR analysis of the aged LmnaSun1 Tmx hearts 12-14 months after Tmx injection confirmed the sustained deletion of Lmna gene (), while protein quantification revealed a significant reduction of LMNA levels in LmnaSun1+Tmx hearts 12-14 months after Tmx (). Histological analysis of the 12-14 month LmnaSun1+Tmx hearts revealed no significant increase in fibrosis compared to controls ( and B). However, echocardiograms on these aged mice showed reduced EF and FS in both LmnaSun1+CTL and LmnaSun1+Tmx mice (), although the average lifespan of Lmnamice is 13-14 months ( and so the reduced contractile function may have been due to ageing. Together these findings demonstrate that loss of Lmna, in adult (2-3 month old) cardiomyocytes is sufficient to result in cardiac failure within 3-4 weeks after Cre activation, but the pathology is strikingly reduced by deleting Sun1, with this reduction being sustained for a year.
Hum Mol Genet
N Engl J Med
Am Heart J
FIG. 26D
FIG. 30A
FIG. 30A
N195K/Flx
+/+
N195K/Flx:mcm
N195K/−
N195K/N195K
N195K/flx:mcm
As most cases of LMNA induced DCM result from missense mutations, we determined what effect loss of SUN1 had on the longevity and cardiac function of a previously described Lmna mutant mouse line carrying the N195K missense mutation that dies from DCM [L. C. Mounkes, et al., 14: 2167-2180 (2005)], with this mutation having been identified in 2 unrelated patients diagnosed with AD-EDMD [D. Fatkin et al., 341: 1715-1724 (1999); J. P. van Tintelen et al., 154: 1130-1139 (2007)]. Here too, we found that the absence of SUN significantly extended the lifespan of this mutant mouse line with improved cardiac function (). We extended these findings by deriving mice heterozygous for the N195K mutation, with the WT-Lmna allele being floxed i.e. Lmna×Sun1. Inducing the Tmx inducible cardiomyocyte Cre allele in these mice (Lmna+Tmx) resulted in the deletion of the WT floxed Lmna allele making the cardiomyocytes hemizygous for the Lmna mutation. These mice had a mean lifespan of less than 50 days, a longevity half that of the original Lmnahomozygotes (). When the Lmna+Tmx mutation was induced on a Sun1 null background longevity was significantly extended from <50 days to >200 days (), revealing that loss of Sun1 is also effective at preventing DCM caused by Lmna missense mutations specifically in cardiomyocytes.
N195K/Flx:mcm
+/+
N195K/Flx:mcm
−/−
N195K/−:mcm
−/−
N195K/−:mcm
+/+
FIG. 30B
FIG. 308
Echocardiograms performed before and after Cre induction revealed progressive worsening of cardiac contractility in the LmnaSun1 mice compared to LmnaSun1 mice (). Loss of SUN1 preserved both EF, FS and Global Longitudinal Strain (GLS) in LmnaSun1 mice compared to LmnaSun1 mice ().
Flx/Flx:mcm
AAV9 Mediated Transduction and Expression of a DNSun1 Prolongs the Lifespan of the Lmna+Tmx Mice.
J Cell Biol
Gene Ther
FIG. 34
FIG. 15
FIG. 31B
The above results demonstrated that genetically ablating SUN1's functions or reducing SUN levels could be of therapeutic value in treating DCM. We then tested whether this was due to the complete ablation of SUN1's functions to overcome its toxic over-abundance versus leaving its levels untouched and specifically disrupting its LINC complex-associated role in tethering KASH-domain proteins in the ONM, thereby tethering the nucleus to components of the cytoskeleton. To distinguish between these 2 possibilities, Adenovirus Associated Virus (AAV) was utilized to transduce and express, specifically in cardiomyocytes, a dominant negative SUN minigene whose protein product would compete with both SUN1- and SUN2-KASH binding in the cardiomyocyte perinuclear space [M. Crisp et al., 172: 41-53 (2006)]. A region corresponding to the entire luminal domain of the Sun1 gene was tagged at its N terminus with an HA (HA-Sun1L) epitope. To localize the resulting protein product to the endoplasmic reticulum (ER) and perinuclear space (between the INM and ONM-PNS), the signal sequence and signal peptidase cleavage site of human serum albumin was fused to the N terminus of HA-Sun1L to yield SS-HA-Sun1L. To prevent the miniprotein's secretion, a KDEL tetrapeptide was linked to the C-terminus of SS-HA-Sun1L, forming SS-HA-Sun1L-KDEL (). The signal sequence would ensure the HA-Sun1KDEL accumulates intracellularly within the contiguous peripheral ER and PNS lumen. The cDNA sequence encoding the minigene was fused to the chicken cardiotroponin promoter (cTnT) to ensure the minigene is only transcribed in cardiomyocytes [K. M. Prasad, et al., 18: 43-52 (2011)]. A diagram of how SS-HA-Sun1L (DN-Sun1) displaces the KASH domain proteins from the LINC complex in the PNS to the ER is presented in (third panel) and .
J Virol.
FIG. 31D
To verify that the DN-Sun1 functioned in cardiomyocytes (CM) we initially transduced human CMs derived from PS stem cells using the AAV-DJ system [D. Grimm, et al., 82(12):5887-911 (2008)] that provides for a higher infectivity rate in cultured cells than the AAV9 serotype used to transduce the DN-Sun1, under transcriptional control of the cTnT promoter, in the mouse hearts. The DN-Sun1 was effective at displacing Nesprin-1 from the nuclear envelopes in the CMs that were expressing the DN-Sun1 as shown in . Cells expressing high levels and low levels of DN-Sun1 are indicated by grey and white arrowheads respectively. High levels of DN-Sun1 expression resulted in the displacement of Nesprin-1 from the nuclear envelope. This confirmed that the DN-Sun1 was effective at disrupting the LINC complex in CMs.
FIG. 31A
FIG. 31C
FIG. 31C
FIG. 38
FIG. 39A
FIG. 39B
FIG. 39C
We used AAV (serotype 9) to transduce and express the DN-Sun1 minigene in the hearts of postnatal mice by intrathoracic injection. The procedure is summarized in and all mice were sacrificed at 100 days after Tmx injection for analysis. Detection by PCR of the Lmna deletion in the hearts confirmed Cre induction by Tmx injection (). To determine the localization and expression levels of the DN-Sun1 minigene, total protein was extracted from half the heart. Western analysis revealed robust expression of both AAV9-DNSun1 and AAV9-GFP control protein (Dose injected: 5×10∧10 vg/g of mouse) 99 days after AAV injection () with the expression levels of both proteins being dependent on the dose of viral particles injected (). The expression of either AAV9-DNSun1 or AAV9-GFP proteins did not affect LMNA protein levels (). Immunofluorescence analysis revealed that a larger percentage of cardiomyocytes were expressing GFP with 5×10∧10 vg/g of AAV9-GFP compared to the levels resulting from a 10-fold lower dose of viral particles (5×10∧9 AAV9-GFP) ( and ).
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flxx/Flx:mcm
Flx/Flx:mcm
Flxx/Flx:mcm
FIG. 20
FIG. 31E
FIG. 31G
FIG. 31G
FIG. 31F
FIG. 31F
The Lmna+Tmx mice, injected with AAV9-GFP control, lived an average of 34.5 days after Tmx, whereas Lmna+Tmx mice injected with AA9-DNSun1 (5×10∧10 vg/g of mouse) lived significantly longer with the majority surviving at least 100 days after Tmx, before their termination for analysis (P=0.0002) ( shows early time period results with male and female mice; shows results at 100 days with separate graphs for male and female mice and mice with different virus injection titre removed). Echo analysis confirmed Lmna+Tmx+AAV9-DNSun1 hearts were functioning better than Lmna+Tmx+AAV9-GFP hearts at 35 days post Tmx (). Although the Lmna+Tmx+AAV9-DNSun1 mice were alive at 100 days after induction both EF % and FS % were significantly lower compared to control Lmna+Tmx mice (). At 35 days post Tmx, increased fibrosis was detected in both the Lmna+Tmx+AAV9-DNSun1 and Lmna+Tmx+AAV9-GFP hearts (), although fibrosis in the Lmna+Tmx+AAV9-DNSun1 hearts was significantly lower than in Lmna+Tmx+AAV9-GFP hearts ( lower panels).
Cell
Cell
Proc Nat Acad Sci USA
PLOS Genet
Mice harboring a variety of Lmna mutations, both global and cardiac-specific, show a significant increase in longevity and health in the absence of Sun1 [(Chen et al., 149: 565-577 (2012) and Examples 2-4]. Prior to the findings described in Examples 2-5, the mechanism of this rescue was unclear, but was speculated to be due to the toxic effects of excess Sun1 in Lmna mutants [Chen et al., 149: 55-577 (2012)]. AAV-mediated expression of a dominant negative LINC-complex-disrupting transgene ameliorates the pathology associated with Lmna mutation [Example 5]. The findings in Examples 2-5 are consistent with the idea that LINC complex function, rather than excess Sun1, is the molecular driver of Lmna pathology. This was surprising, as genetic disruption of the LINC complex via loss of Sun1 and Sun2 [K. Lei at al., 106: 10207-10212 (2009)], or cardiac-specific disruption of Nesprin-1 and Nesprin-2 [Banerjee at al., 10(2): e1004114 (2014)], in mice, resulted in various pathologies.
Current topics in developmental biology
Biol. Chem.
To develop alternative means of disrupting the LINC complex in vivo, the possibility of using CRISPR/Cas9 genome editing to disable the SUN and KASH domains of the proteins constituting the LINC complex was examined. As both the SUN domain and the KASH domain are located at the C-termini of their respective proteins, we hypothesized that a CRISPR guide RNA targeted to the 3′ end of the genes encoding SUN or KASH domain proteins would result in a premature stop codon following CRISPR-induced non-homologous end joining. This would result in a truncated protein with its C-terminal SUN or KASH domain mutated. The truncated protein would be expressed and membrane-localized, but unable to interact with its cognate LINC complex partners. In Example 2, we found that loss of Sun2 did not ameliorate Lmna-associated pathologies. Thus we chose to target the Sun1 SUN domain using CRISPR as Sun1 appears to be the dominant SUN domain protein mediating Lmna pathology. Of the KASH domain proteins, only Nesprin-1, Nesprin-2 and Nesprin-3 are broadly expressed [H. F. Horn, 109: 287-321 (2014)). Nesprin-1 and Nesprin-2 are close paralogues that are functionally redundant. They interact with the actin and microtubule cytoskeleton, whereas Nesprin-3 appears to interact specifically with intermediate filaments [Kim et al., 396: 295-310 (2015)]. As we already had Nesprin-2 and Nesprin-3 mutant mouse strains derived by conventional gene targeting available in the laboratory, we chose to target the KASH domain of Nesprin-1 using CRISPR to test the possibility of using CRISPR/Cas9 in vivo for treatment of laminopathies.
The Sun1 gene and the Syne1 gene encoding Nesprin-1 protein were directly targeted in vivo by microinjecting C57/B16 mouse zygotes with Cas9 mRNA and gRNA targeting the SUN1 (5′-GCACAATAGCCTCGGATGTCG-3′; SEQ ID NO: 66) or KASH1 (5′-CCGTTGGTATATCTGAGCAT-3′ SEQ ID NO: 67) domains, followed by implantation into surrogate mothers. Note the SUN gRNA targeted Sun1 upstream of the SUN domain so as to ablate the SUN domain. A gRNA (5′-GGTTATGGCCGATAGGTGCAT-3′; SEQ ID NO: 68) targeting the tyrosinase gene was co-injected—progeny that had undergone CRISPR genome editing would have white or mosaic coat color resulting from tyrosinase disruption. These pups were genotyped to confirm successful gene disruption and used as founder animals to establish Sun1 or Nesprin-1 mutant colonies.
FIG. 40A
FIG. 41A
FIG. 40B
FIG. 40C
Following Sanger sequencing of founder animals and F1 progeny, we focused on characterizing Sun1 mutant alleles () with a 7 bp deletion (Sun1_del7, or Sun1Δ7; SEQ ID NO: 71) and 4 bp insertion (Sun1_plus4; SEQ ID NO: 70), and a Syne1 (Nesprin-1) mutant allele () with a 8 bp deletion (Syne1_CTdel8, or Syne1 C′TΔ8; SEQ ID NO: 78). The Sun1 mutant alleles were predicted to produce mRNA with premature stop codons resulting in a truncated Sun1 protein lacking a SUN domain (). Tail tip fibroblasts were isolated from Sun1 homozygous mutant animals. Immunofluorescence staining revealed loss of Sun1 protein (), suggesting that the indels generated by CRISPR caused nonsense-mediated decay of Sun1 mRNA. It is unclear whether the site of mutation, being outside the SUN domain rather than inside the SUN domain, had an effect on the expression of the mutated gene. As we were unable to obtain Sun1 mutant alleles that produced Sun1 protein lacking the SUN domain, instead obtaining essentially Sun1 null animals, we did not further characterize these mutant lines.
FIG. 41B
FIG. 41C
FIG. 41C
The Syne1 C′TΔ8 allele is predicted to produce a protein where the final 11 amino acids in the wildtype sequence (SEQ ID NO: 77) are mutated and are followed by an additional 50 amino acids encoded by an alternate reading frame (; SEQ ID NO: 78). Immunoblotting performed on Syne1WT and Syne1C′TΔ8 heart and muscle tissues revealed a ˜120 kDa band in WT corresponding to the striated muscle-enriched Nesprin-1α isoform of the Syne1 gene (, D). In the C′TΔ8 heart and muscle tissues, the presumptive Nesprin-1a polypeptide appeared to be less abundant and of lower electrophoretic mobility than in the wildtype (, D). This is consistent with the 8 bp deletion in the Syne1C′TΔ8 allele introducing a novel stop codon downstream, resulting in a protein of higher molecular weight. In addition, a ˜1 MDa band likely corresponding to Nesprin-1Giant was observed in heart tissue from both Syne1WT and Syne1C′TΔ8 mice.
FIG. 42A
FIG. 42B
FIG. 43A-C
FIG. 44A-C
Curr. Biol.
Immunofluorescence analysis of mouse adult fibroblasts (MAFs) derived from 12 week old mice revealed that Nesprin-1 was mis-localized from the nuclear envelope to the cytoplasm in the Syne1C′TΔ8 MAFs (). Similarly, in myotubes, Nesp-1 redistributes to the cytoplasm in the Syne1C′TΔ8 myotubes as compared to Syne1WT (). Other LINC complex and NE proteins such as SUN, SUN2, Emerin and LaminA remained localized to the NE (). Consistent with previous reports [Gimpel at al., 27: 2999-3009.e9. (2017)], disruption of Nesprin-1 in myotubes led to mislocalization of centrosomal proteins PCM1. Pcnt and Akap450 from the myotube nuclear envelope (). Mislocalization of Nesprin-1 from the nuclear envelope is consistent with disruption of the Nesprin-1 KASH domain, preventing Nesprin-1C′TΔ8 mutant protein from interacting with the SUN domains of Sun1 and Sun2, which would normally restrict Nesprin-1 to the nuclear envelope. As the transmembrane region is not disrupted, it is likely that Nesprin-1 is mislocalized to the endoplasmic reticulum (ER) in the C′TΔ8 mutant, as the ER and the perinuclear space form a contiguous membrane system.
Hum Mol Genet
Hum Mol Genet
FIG. 45A-B
FIG. 45C
Similar to one previously reported Nesprin-1 mouse model [Zhang et al., Development 134(5): 901-8 (2007)], and in contrast to two other models [Puckelwartz at al., 18: 607-620 (2009); Zhang et al., 19: 329-341 (2010)], the disrupted KASH domain of Nesprin-1 results in no overt phenotypic differences between the Syne1 wildtype (WT) and Syne1C′TΔ8 mutant (). Both male and female homozygous mutants were fertile with no significant differences in body weight between the Syne1WT and Syne1C′TΔ8 mice (). Syne1C′TΔ8 mice also did not exhibit any growth retardation or obvious muscle dystrophy, nor did they display any difficulty in movement or grooming, which can be indications of muscle deterioration.
FIG. 46A
FIG. 46B
FIG. 46C
Development
−/−
C′TΔ8/C′TΔ8
−/−
In order to probe the role of other KASH domain proteins in Lnma pathology, mice mutant for Syne2, encoding Nesprin-2, were generated by conventional gene targeting (). To characterize the mutation, immunofluorescence microscopy of tail tip fibroblasts was carried out. Syne2+homozygous mutant fibroblasts expressed little to no Nesprin-2 (). Consistent with previous findings [Zhang at al., 134(5): 901-8 (2007)], while Syne2 mice were overtly normal, with no growth retardation or infertility, Nesprin-1/2 double mutant mice (Syne1:Syne2) were perinatal lethals ().
C′TΔ8/C′TΔ8
Δ/Δ
Δ/Δ
C′TΔ8/C′TΔ8
Δ/Δ
Δ/Δ
C′TΔ8/C′TΔ8
C′TΔ8
Δ/Δ
−/−
FIG. 24
FIG. 47
Even though Nesprin-1 was still expressed, Nesprin-1-containing LINC complexes would not be formed in Syne1cells and animals. Since AAV-mediated disruption of the LINC complex using dominant negative Sun1 in vivo rescues Lmna pathologies (Example 5), we reasoned that the “KASH-less” Nesprin-1 mutant allele we generated might also rescue Lmna pathology. To test this hypothesis, mice heterozygous for a Lmna null (Lmna) allele (Example 1) were intercrossed with Syne1C′TΔ8 mice to obtain Lmna:Syne1double mutant mice. While Lmna mice lived for 15-17 days, Lmna:Syne1double mutant mice lived for up to 42 days (). Lmna null mice heterozygous for the Syne1allele did not experience any lifespan extension. Lmna mice on a Syne2 homozygous mutant background also did not experience lifespan extension (), indicating that Lmna pathology is mediated primarily by Nesprin-1/Sun1 LINC complexes.
C′TΔ8/C′TΔ8
Flx/Flx
C′TΔ8/C′TΔ8
FIG. 25
To examine the effect of the Syne1allele in mice with cardiac-specific loss of Lmna, mice homozygous for a conditional Lmnaallele carrying the inducible cardiomyocyte specific Cre Tg(Myh6-cre/Esr1) (here abbreviated to mcm), in which Cre is induced by a single injection of tamoxifen (Tmx), were used as described in Examples 1-2. Cardiac-specific deletion of Lmna results in death within a month, but mice with the same deletion induced on a homozygous Syne1background lived for at least 120 days after Tmx induction (; no change from day 80-120).
Method for Screening Small Molecules that Block SUN-KASH Interactions
Cell
Cell
Cell
Cell
FIG. 5B
FIG. 21
FIG. 21
FIG. 22
Crystallographic studies of human SUN2 reveal that the SUN domain is assembled as a clover-like trimeric structure [Sosa et al., 149(5): 1035-47 (2012)]. Trimerization is mediated by a triple-helical coiled-coil, with an estimated length of 40-45 nm. This is sufficient to bridge the perinuclear space (PNS), allowing SUN and KASH domains to directly interact [Sosa et al., 149(5): 1035-47 (2012)]. The KASH binding site is formed primarily within a groove formed at the interface between adjacent SUN domains (; left panel). This groove accommodates part of the KASH domain, about 18 residues, in an extended conformation. However, it is the C-terminal tetrapeptide of the KASH domain, featuring three proline residues followed by a terminal aliphatic reside, Leu or Thr (for Nesp1 and Nesp2 respectively), that is crucial for the SUN-KASH interaction ( right panel, adapted from FIG. 1 of Sosa et al, 149(5): 1035-47 (2012). The significance of this tetrapeptide is that it is situated in a well-defined pocket formed within a single SUN monomer. Modification of this peptide in any way, including the addition of a single residue (an Ala) at the C-terminus, completely eliminates the SUN-KASH association over the entire SUN-KASH contact region (Sosa et al., 149(5): 1035-47 (2012) and left panel). The conclusion is that while stable binding of the KASH domain requires 18-20 residues, it is the C-terminal tetrapeptide that actually initiates binding. Thus, blocking the tetrapeptide binding-pocket within the SUN monomer will abolish SUN-KASH association. We have described in this disclosure an AAV-based gene therapy strategy to break endogenous SUN-KASH interactions as a treatment for laminopathies, including dilated cardiomyopathy. Alternatively, a small molecule that blocks the SUN-KASH interaction at the SUN binding pocket would disrupt LINC complexes and similarly treat laminopathies. A variety of standard methods exist to screen for small molecule drugs in vitro.
Cell
Cell.
FIG. 22
An in vitro screen can be set up employing recombinant SUN and KASH domains or KASH peptide, for which methods of production have been previously published [Sosa at al., 149(5): 1035-47 (2012)]. One such screen involves an assay technique analogous to an enzyme-linked immunosorbent assay ( right panel, similar to Lepourcelet et al., Cancer 5(1):91-102 (2004)). Recombinant SUN domain is immobilized on a solid surface, typically in 98-well plates, and then complexed with recombinant KASH domain linked to an enzyme that can generate a colorimetric or chemiluminescent readout. One method for enabling this linkage is to synthesize a biotinylated KASH peptide, which can then be linked with commercially available streptavidin-horseradish peroxidase (HRP) conjugate. Candidate compounds are obtained from appropriate suppliers and screened for their ability to inhibit KASH-SUN associations in vitro. Compounds that fail to inhibit the SUN-KASH interaction will result in a well in the plate where the recombinant SUN binds to the enzyme-linked KASH domain. Following wash steps and incubation with colorimetric or chemiluminescent HRP substrates, the presence of the SUN-KASH interaction is detected in standard plate readers. If the compound can inhibit SUN-KASH interaction then, following the wash step, the KASH domain is removed and there would be reduced or no enzymatic reaction in the well.
Expert Opin Drug Discov
Alternatively, fluorescence anisotropy or polarization can be used to screen for small molecule inhibitors of SUN-KASH interactions in vitro [Lea, W. A., and Simeonov, A. 6: 17-32 (2011)]. This assay also employs recombinant SUN and KASH domains. The KASH domain is fluorescently labeled; for example a chemically synthesized KASH peptide could be readily functionalized with a fluorescein moiety. Fluorescence anisotropy of the interacting KASH domain interacting with SUN domain can be measured using standard equipment such as a plate reader. A small molecule inhibitor that disrupts the SUN-KASH interaction can be readily detected as the fluorescence anisotropy of the fluorescent KASH will change if it is not bound to SUN.
FIG. 23
As is typical in drug screening campaigns, the compounds which successfully pass the in vitro primary screen will then be subjected to cell-based secondary screens (). In this case, immunofluorescence microscopy will be employed to identify those compounds that can dissociate LINC complexes. This is manifest as dispersal of the KASH component to the peripheral endoplasmic reticulum while the cognate SUN protein is retained in the inner nuclear membrane. This microscopy-based assay can be performed first on HeLa cells. Active compounds are then evaluated on cultured cells from disease-relevant tissue, such as cardiac cells. An additional secondary screen may include the ability of the identified compound to rescue proliferation defects in Lmna knockout cells. Following hit-to-lead optimization of the identified compound using standard methods, the compound can be tested in mouse models of laminopathies such as those described herein for Lmna dilated cardiomyopathy. Efficacy of the leads can be evaluated using lifespan of the mutant mice and echocardiograms, as described herein, to assess heart function.
J Am Coll Cardiol
J Am Coll Cardiol
DCM caused by LMNA is regarded as being aggressive, and often leads to premature death or cardiac transplantation [M. Pasotti et al., 52: 1250-1260 (2008); M. R. Taylor et al., 41: 771-780 (2003)]. By 60 years, 55% of LMNA mutation carriers die of cardiovascular failure or receive a heart transplant, compared with 11% of patients with idiopathic cardiomyopathy. Attempts to ameliorate DCM by fitting a pacemaker have been at best of transient benefit. Consequently it is necessary develop new therapeutic avenues to treat DCM caused by LMNA mutations.
Nature genetics
Science translational medicine
Science translational medicine
Circulation
The majority of LMNA mutations causing DCM are dominant negative missense. Treatment by conventional gene therapy to repair each mutation would be daunting and removal of the mutated allele, leaving the patient hemizygous for the remaining normal WT allele may also result in heart failure [G. Bonne et al., 21: 285-288 (1999)]. Various other routes downstream of the Lamin gene have been explored for potential therapeutic intervention, and have included mTOR inhibition with rapamycin/rapalogues [J. C. Choi et al., 4: 144ra102 (2012); F. J. Ramos et al., 4: 144ra103 (2012)] and inhibition of the MEK1/2 kinase pathway [W. Wu, et al., 123: 53-61 (2011)]. Both avenues, resulted in improved ventricular function and increased longevity (10-40%) but the extent and long-term efficacy was significantly less than that we observed with the loss of Sun1.
Science translational medicine
Dev Cell
Hum Mol Genet
Hum Mol Genet
Hum Mol Genet
Front Physiol
Hum Mol Genet
Cell
The molecular mechanisms underlying the varied phenotypes of the laminopathies are still not well understood, though two alternative hypotheses have been proposed to explain the tissue-specific pathologies. The first “gene regulation hypothesis” proposes that LMNA mutations/loss disrupt the equilibrium of various molecular pathways due to the mutations altering interactions with NE proteins and chromatin, which in turn alter gene expression. Evidence in support of this hypothesis comes from studies reporting changes in signalling pathways including the AKT-MTOR pathway [J. C. Choi et al., 4: 144ra102 (2012)], WNT/β-catenin pathway [L. Hernandez et al., 19: 413-425 (2010); C. Le Dour et al., 26: 333-343 (2017)], TGF-β/Smad [J. H. Van Berlo et al., 14: 2839-2849 (2005); T. V. Cohen et al., 22: 2852-2869 (2013)], MAP Kinase pathway [A. Brull, et al., 9: 1533 (2018)] and the ERK1/2-CTGF/CCN2 pathway [M. Chatzifrangkeskou et al., 25: 2220-2233 (2016)]. While these changes have been documented, none has clearly established whether these changes are not a secondary compensatory effect of a diseased tissue. Sun1 also fits into this rubric of disrupted expression levels as Sun1 protein, but not mRNA, is upregulated in laminopathies, leading to the proposal that laminopathy phenotypes are caused by toxicity from excess Sun1 [C. Y. Chen et al., 149: 565-577 (2012)].
J Clin Invest
Physiol Rev
J Clin Invest
J Cell Biol
J Biol Chem
J Biol Chem
Proc Natl Acad Sci
Development
PLOS Genet
FIGS. 28 and 29
FIG. 32A-C
The second hypothesis suggested Lmna loss or mutation leads to Increased nuclear fragility. As a result mechanical stress and tension forces transmitted via the LINC complex from the cytoplasm to the NE causes damage to the NE [J. Lammerding et al., 113: 370-378 (2004)]. This hypothesis is similar to that proposed for Duchenne muscular dystrophy (DMD), where loss of dystrophin increases the fragility of the muscle cell membrane and when tension-stress forces are applied during muscle contraction this results muscle cell rupture and death [D. J. Blake, et al., 82: 291-329 (2002)]. Lmna mutant fibroblasts show nuclear deformation, defective mechanotransduction, and reduced viability when subjected to mechanical strain, together with increased nuclear rupture at low and moderate pressures when compared to WT nuclei [J. Lammerding et al., 113: 370-378 (2004); J. Lammerding et al., 170: 781-791 (2005); J. Lammerding et al., 281: 25768-25780 (2006)]. In contracting mouse cardiomyocytes, mechanical stress and tension forces caused by 500-600 contractions per minute are transmitted to the NE via the LINC complex, resulting in nuclear distortion, damage and eventual death/loss as described in . Presumably, such forces would cause significant damage to the fragile NE of Lmna null cardiomyocytes, resulting in CM death. If the tension-stress hypothesis is damaging to the NE, then unlinking the LINC complex, by disrupting SUN1, would reduce the tension-stress on the CM nuclei, and prevent CM cell death in the mutant CMs (). One caveat here is that complete disruption of the LINC complex, as would be the case following overexpression of DN-Sun1, could potentially be deleterious rather than therapeutic. At the cellular level, multiple mechanical phenomena including intracellular force transmission, cell polarization and migration, were impacted following LINC complex disruption by dominant negative SUN and KASH constructs [Lombardi et al., 286(30):26743-53 (2011)]. In animal models, Sun1/Sun2 [Lai et al., 106(25):10207-12 (2009)] and Nesprin-1/Nesprin-2 [Zhang et al., 134(5):901-8 (2007)] double mutant mice experience perinatal lethality and cardiac-specific disruption of the KASH domains of Nesprin-1 and Nesprin-2 using an embryonic cardiac Cre driver (Nkx2.5-Cre) results in early onset cardiomyopathy [Banerjee et al., 10(2):e1004114 (2014)].
FIGS. 32D & 34
FIG. 31C
FIG. 31C
FIG. 31D
FIGS. 31E
Mol Ther
We attempted to distinguish the tension-stress hypothesis from the expression level hypothesis in cardiomyocytes, using a DN-Sun1 construct to compete with endogenous Sun1 and Sun2 proteins for KASH-domain-binding and so unlink the LINC complex without directly altering Sun1 levels (). The AAV9 vector, which has a high affinity for CM, was used to deliver DN-Sun1 under the cTnT promoter to CMs [C. Zincarelli, et al., 16: 1073-1080 (2008)]. Our results showed the successful delivery of GFP to cardiomyocytes (), and robust expression of both the control GFP and DN-Sun1 proteins () with the latter resulting in the dispersal of the KASH domain proteins from the cardiomyocyte nuclei (). Surprisingly, not only did AAV-DN-Sun1 ameliorate the pathology in mice with depleted cardiac Lmna levels, it also had no discernible effect on the cardiac health of wildtype mice, which would be expected to also experience complete LINC complex disruption in their hearts ( & G). This suggests that an intact LINC complex may be required in embryonic development, but not postnatally.
C′TΔ8/C′TΔ8
In addition, using CRISPR/Cas9 in nice, we generated a Syne1 mutant allele (C′TΔ8) that gave rise to a truncated Nesprin-1 protein with a disrupted, non-functional, KASH domain. Mice lacking Lmna globally or in the heart have a shortened lifespan, but the presence of a homozygous Syne1mutation resulted in significant lifespan extension. Loss of Sun1 or AAV-mediated disruption of the LINC complex by dominant negative transgenes in vivo resulted in similar rescue of Lmna pathology (Example 2-5), while Sun2 and Nesprin-2 mutations did not. Taken together, these data suggest that LINC complexes comprised of Sun1 and Nesprin-1 drive the pathology in Lmna mutant cells and animals.
Staphylococcus aureus
Streptococcus pyogenes
Cell
There have been a number of reports on the use of AAV to deliver CRISPR/Cas components in vivo for treating diseases. Our results predict that AAV-mediated CRISPR/Cas, such as CRISPR/Cas9, delivery to target the Nesprin-1 KASH domain in disease-affected tissue can be used to treat laminopathies, including dilated cardiomyopathy. For instance, cardiotropic AAVs (e.g. AAV9) can be used to deliver transgene cassette(s) containing a cardiac-specific promoter (e.g. cTnT) driving Cas endonuclease enzyme expression and an appropriate promoter (e.g. U6) driving gRNA expression to treat LMNA DCM. Since the packaging capacity of AAV is limited to 4.7 kb, a smaller Cas9 derived from (saCas9) rather than the larger, more commonly used, Cas9 may be preferred. Alternatively, other CRISPR enzymes such as Cpf1, which is small enough for AAV packaging and has a more commonly found protospacer adjacent motif (PAM) than saCas9, could be used [Zetsche, B., et al., Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. 163, 759-771 (2015)].
Cell
Δ/Δ
−/−
Δ/Δ
Guide RNAs would target the 3′ region of the Nesprin-1 gene encoding the KASH domain (Table 3). While we have targeted the region adjacent to the stop codon, in principle any gene region encoding the KASH domain could be targeted as indels generated by CRISPR would likely result in frameshift mutations that disable the KASH domain. However, as the final 4 amino acids in the KASH domain are known to be absolutely required for SUN domain interaction and hence LINC complex formation [Sosa et al., 149(5):1035-47 (2012)], it is prudent to select gRNA in the vicinity of the stop codon as even indels that do not result in a frameshift could still mutate the relevant KASH amino acids required for the SUN-KASH interaction. Furthermore, it should be noted that because the Syne1 gene encoding Nesprin-1 is very large and has multiple splice isoforms and alternative start sites, guide RNAs targeted outside the KASH domain, while giving rise to some mutant Nesprin-1 isoforms, may not perturb expression of other isoforms of Nesprin-1 protein, including KASH-containing isoforms. This would result in formation of functional or partially functional Nesprin-1/Sun1 LINC complexes that would still be able to drive pathology in Lmna mutants. This CRISPR/Cas9 strategy likely cannot be extended to the KASH domain of Nesprin-2, since Lmna:Syne2 mice are phenotypically indistinguishable from Lmn mice.
Cell
PLOS Biology
MBoC
Cell
We did not further investigate the Sun1 mutant mice generated in this study as instead of mice with Sun1 lacking the SUN domain, we essentially obtained Sun1 null mice, which have already been well characterized. We suspect that inducing CRISPR mutation in Sun1 resulted in nonsense-mediated decay (NMD) of Sun1 transcript. Occurrence of a premature termination codon (PTC) 50-55 nucleotides upstream of a exon-exon junction is a trigger for NMD [Popp, M. W., and Maquat, L. E. 165: 1319-1322 (2016)]. Also, PTCs occurring in the middle of a transcript are more likely to result in NMD [Eberle et al., 6: e92 (2008); Reber et al., 29: 75-83 (2018)]. In the Sun1_plus4 mutant, the PTC is more than 55 nucleotides upstream of the exon-exon junction and is likely to trigger NMD. For the Sun1Δ7 mutant, the PTC is less than 50 nucleotides from the exon-exon junction. However for both mutants, since we targeted upstream of the sizeable SUN domain, the PTCs are roughly ⅔ of the way along the length of the transcript, and hence also likely to trigger NMD. In order to specifically disrupt the SUN domain in Sun1 without inducing a null mutation, we can adopt a similar strategy as for Nesprin-1—directing the guide RNA at the very 3′ end of the coding region of the transcript (Table 3). Earlier work demonstrated that mutation of a tyrosine residue to phenylalanine at the C-terminus of SUN2 (Y707F) abolished KASH binding [Sosa et al., 149(5):1035-47 (2012)]. This critical tyrosine residue is conserved in SUN1 (Y812 in Uniprot E9PHI4) and present in the final coding exon of the SUN transcript. Selection of a gRNA 5′ proximal to the codon for Y812 would produce Indel mutations that cause a frameshift mutation that would mutate Y812 and disrupt KASH binding. As the gRNA would be in the final coding exon, the likelihood of triggering NMD would be low. One can thus envision a CRISPR/Cas9-based strategy to treat laminopathies by targeting a critical residue required for KASH-binding in the SUN1 SUN domain. AAVs could be used to deliver CRISPR enzyme and gRNA targeting SUN in appropriate disease tissue, such as the heart. Incapacitation of SUN KASH binding would then ameliorate the deleterious effects of Lmna mutations.
From these results we propose that the loss of or mutations within Lmna causes instability in the CM nuclei due to loss or incorrect assembly of the nuclear lamina. This makes the nuclei susceptible to the tension/stress forces exerted via the LINC complex from the contractile sarcomeres of the CMs. In the absence of SUN1, or following mutation of Nesprin-1 KASH domain, the untethered LINC complexes exert less tensional force on the CM nuclei, enabling survival of the lamin deficient cardiomyocyte.
Δ/Δ
−/−
These results provide an opportunity to use the AAV-mediated delivery of DN-Sun, DN-KASH, or direct mutation of endogenous SUN or KASH proteins as potential therapeutics for laminopathy-related DCM in patients. The AAV system, as a therapeutic delivery route in patients is established and has been approved by the FDA for treating some diseases. It is becoming more widely used with multiple on-going clinical trials, including the introduction into patients with heart disease. However, even though tension-stress may be the primary cause for Lmna deficient CM death, disrupting SUN1 may not be effective in preventing LMNA mutation induced cell death in skeletal muscle, as Lmna:Sun die at an earlier age than those mice where Lmna was specifically deleted in the CMs. Which muscle groups (or even other tissues lacking Lmna) result in the early lethality remain to be identified. However in most of the LMNA DCM patients it is heart failure that is the cause of death, and our results show that disrupting the LINC complex in CMs could be effective at preventing heart failure for an extended period.
Circulation Research
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70. Zhang, X., Xu, R., Zhu, B., Yang, X., Ding, X., Duan, S., Xu, T., Zhuang, Y., and Han, M. (2007). Syne-1 and Syne-2 play crucial roles in myonuclear anchorage and motor neuron innervation. 134: 901-908.
Mol Ther
71. C. Zincarelli, S. Soltys, G. Rengo, J. E. Rabinowitz, Analysis of AAV serotypes 1-9 mediated gene expression and tropism in mice after systemic injection. 16: 1073-1080 (2008).
BRIEF DESCRIPTION OF THE FIGURES
FIG. 1
shows a schematic of the mutations in the lamin A/C gene LMNA and the laminopathies resulting from the mutations.
FIG. 2
shows a schematic of the positioning of components of the nuclear envelope membrane and lamina.
FIG. 3
shows a schematic of the connections between the nucleus and the extracellular matrix via the LINC complex and how mutations in lamin A/C might result in DCM. The plasma membrane, cytoskeleton and nucleus form a mechanically and physically linked entity. In Lmna mutants, the nucleus is structurally weak. It is much more susceptible to mechanical stress from cytoskeletal forces. This leads to severe damage to the myocyte nuclei that in turn leads to a cascade of events such as apoptosis and fibrosis that results finally in DCM.
FIG. 4
−/−
−/−
shows the effect of microinjection of dextran into the nucleus of Lmna and Lmna mice under low pressure. In the wildtype cells, the dextran stays in the nucleus, while in the Lmna mutant cells the dextran leaks out of the nucleus into the cytoplasm.
FIGS. 5A-5B
FIG. 5A
FIG. 5B
show schematics of a LINC complex () and Interaction between KASH and SUN ().
FIG. 6
−/−
−/−
−/−
−/−
−/−
−/−
−/−
−/−
−/−
−/−
+/−
−/−
−/−
−/−
−/−
−/−
−/−
+/+
−/−
−/−
+/+
+/−
+/+
−/−
−/−
+/+
−/−
shows defects in body weight and longevity in Lmna and LmnaΔ9 mice are ameliorated in homozygous Sun1 knockout Lmna Sun1 and LmnaΔ9Sun1 animals. (A) Body weights are averages from mice with the indicated genotypes. The number (n) of animals used Is Indicated. (B) Kaplan-Meler graph showing Increased life span of LmnaSun1 compared to Lmna mice. Median survival of wild-type or Sun1 is >210 days in a 7 month follow up; Lmna mice have median survival of 41 days; LmnaSun1 mice have a median survival of 54 days; Lmna Sun1 mice have a median survival of 104 days (p<0.01 comparing Lmna and Lmna Sun1). (C) Body weights of LnmaΔ9 mice that are wild-type, heterozygous, or homozygous for Sun1 deficiency. Wild-type and Sun1 cohorts are graphed for comparison. Values are averages t SEM from animals in each cohort. Number (n) of animals is indicated (p<0.0001 comparing LmnaΔ9Sun1 and LmnaΔ9Sun1). (D) Kaplan-Meier graph showing increased life span of LmnaΔ9Sun1 compared to LmnaΔ9Sun1 mice. LmnaΔ9Sun1 mice are also graphed. (p<0.0001 comparing LmnaΔ9Sun1 and LmnaΔ9Sun1). (E) Cell proliferation of the indicated MEFs. Curves are averages±SD, representative of >3 independent isolates from embryos of the indicated genotypes. (F) Proliferation curves of MAFs (mouse adult fibroblasts) from WT. Sun1, LmnaΔ9Sun1 and LmnaΔ9Sun1 mice. MAFs were seeded at a density of 1000 cells per well. Growth was measured, and normalized cell indexes (averages±SD) are presented.
FIG. 7
shows a schematic of the features of the Sun1 protein and the components used to generate a dominant negative Sun1 protein, including a signal sequence, oiled-coil sequence, SUN domain sequence and KDEL sequence.
FIG. 8
shows a schematic of a plasmid (SEQ ID NO: 1) used for AAV production.
FIG. 9
shows a schematic of a plasmid (SEQ ID NO: 2) comprising sequences from AAV2 and AAV9 for AAV production.
FIG. 10
shows a schematic of an AAV expression construct (SEQ ID NO: 3) comprising cardiac-specific promoter and Sun1 dominant negative sequence.
FIG. 11
shows a schematic of the features of the dominant negative Sun1 protein, Including a signal sequence, coiled-coil sequence, SUN domain sequence and KDEL sequence (SEQ ID NO: 4).
FIG. 12
shows a schematic of the features of a dominant negative Sun2 protein, including a signal sequence, lumenal domain sequence and KDEL sequence (SEQ ID NO: 5).
FIG. 13
shows a schematic of the region of Sun1 protein used in dominant negative constructs.
FIG. 14
shows an alignment of KASH1-KASH5 domain amino acid sequences with conserved residues (SEQ ID Nos: 7, 9, 11, 13 and 15, respectively).
FIG. 15
shows a schematic of the LINC complex in wildtype mice, Sun1 KO mice, AAV dominant negative SUN mice and mice with altered KASH domain. The schematic for wildtype mice is obtained from Brian Burke, 2012. The schematic for Sun1 KO mice represents the results from Chen et al., 2012. The AAV dominant negative SUN and the altered KASH domain schematics represent Inventor proposals at the priority date on methods for LINC complex disruption to ameliorate laminopathies, based on data obtained at that time.
FIG. 16
shows a Kaplan Meier curve of Lmna KO mice surviving for an average of 28 days, Sun1 KO mice living beyond 300 days and cardiac Lmna KO/Sun1 KO mice living beyond 300 days.
FIG. 17
shows H&E stained sections of hearts from Sun1 KO mice, cardiac Lmna KO mice and cardiac Lmna KO/Sun1 KO mice, with LmnaKO/Sun1WT hearts showing enlargement of the left ventricle (DCM) compared to WT and LmnaKo/Sun1KO hearts.
FIG. 18
shows a schematic of disruption of a LINC complex in a Nesprin-1 ΔKASH mouse. LmnaKO Nesprin-WT mice have a lifespan of about 20 days. LmnaKO Nesprin-1-ΔKASH survive about 40 days, which is similar to LmnaKOSun1KO mice.
FIGS. 19A-19B
FIG. 19A
FIG. 19B
show a schematic of anticipated AAV-cTNT-DN-SUN expression and competition between exogenous DN-SUN and native Sun1 for binding to the KASH domain () with the DN-SUN shown in 19B (upper panel) and the effect of transfected DN-SUN on native Nesprin2G positioning in cells where the 2 nuclei in the middle panel express the DN-SUN and in the merge panel both show loss of Nesprin2 from the nuclear membranes ().
FIG. 20
is a Kaplan Meier curve showing disruption of SUN-KASH interaction in vivo, using AAV9-cTNT-dominant negative Sun1 (DNSun1), extends the longevity of the heart-specific Lmna KO in male and female mice.
FIG. 21
shows C-terminal amino acids of the KASH domain of Nesprin-2 (KASH2). The 14 or 18 amino acid sequence from KASH2 C-terminus are able to physically interact with the SUN domain of SUN2. Loss of the last 4 amino acids from KASH2 or addition of a single alanine amino acid at the C-terminus of KASH2 is sufficient to disrupt interaction of the KASH2 domain with the SUN domain.
FIG. 22
shows a schematic of a screening method for detecting agents that disrupt the LINC complex.
FIG. 23
shows a flowchart showing a more detailed screening method for identifying a small molecule to disrupt the LINC complex.
FIG. 24
Δ/Δ
+/+
+/C′TΔ8
Δ/Δ
C′TΔ8/C′TΔ8
is a Kaplan Meier curve showing that wild type (C57/Bl6) mice with or without a Nesprin-1 KASH-disrupting (C′TΔ8) mutation have a normal lifespan. Mice with a Lmna null/KO mutation (LA-ZP3cre) and wildtype (Nesp1) or heterozygous (Nesp1) for Nesp1-C′TΔ8 have a median lifespan of 15 or 18 days, which is increased to 38 days in Lmna KO/homozygous Nesp1 mutant (LA-ZP3cre; Nesp1) mice.
FIG. 25
CTΔ8/CTΔ8
+/+
+/−
CTΔ8/CTΔ8
f/f
+/+
WT/WT
f/f
+/+
WT/WT
f/f
+/−
CTΔ8/CTΔ8
f/f
+/−
is a Kaplan Meier curve showing that mice with wildtype Lmna (N1LAMCre), or floxed alleles of Lmna but lacking a cardiac-specific Cre driver (N1LAMCre and N1LAMCre), live for length of the experiment (˜80 days at priority filing, which extended to 120 days unchanged). Mice with a cardiomyocyte-specific deletion of Lmna (N1LAMCre) have a lifespan of 22-24 days following induction of the Cre/loxP-mediated deletion by tamoxifen (TMX) delivery, which is increased to the length of the experiment in mice with a cardiomyocyte-specific deletion of Lmna induced by TMX and also homozygous mutant for Nesprin-1 (N1LAMCre).
FIGS. 26A-26D
FIG. 26A
FIG. 26B
FIG. 26C
FIG. 26D
Flx/Flx:Zp3
−/−
Flx/Flx
αMyHC
Flx/Flx:αMyHC
−/−
Flx/Flx
−/−
N195K/N195K
N195K/N195K
−/−
show Kaplan Meier curves of Sun1 loss extending the longevity of Lmna mutant mice. () Wild type (C57/Bl6) mice with or without Sun1 have a normal lifespan, whereas the average postnatal lifespan of the Lmnamice in which LaminA is deleted in all tissues was 17.5 days (***P=<0.0001; Log-rank test). On a Sun1 background longevity is increased to 32.5 days. () When Lmnawas deleted specifically and constitutively in hearts by crossing the mice with the Creline, the Lmnamice lived on average 26.5 days. On a Sun1 background these mice lived for longer than 6 months. () 3-5 month old Lmnawere crossed with the Tmx inducible cardiomyocyte specific Cre Tg(Myh6-cre/Esr1), (abbreviated to mcm), after a single injection of Tmx the mice die within 3-4 weeks. On a Sun1 background these mice lived for more than 1 year. () Lmnamice lived for an average of 78 days compared to LmnaSun1 mice which had an average lifespan of 111 days. (***P=<0.0001, **P=0.0073 Log-rank test).
FIGS. 27A-27E
FIG. 27A
FIG. 27B
FIG. 27C
FIG. 27D
FIG. 27E
FIG. 27F
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx
Wt/Wt
show the lifespan and phenotype of Lmna+Tmx mice. () The average lifespan of the Lmnamice was 27 days after a single Tmx injection (***P=<0.0001; Log-rank test). () PCR detected the floxed (deleted) Lmna gene (arrow head) only in heart tissue after Tmx injection and not in other tissues or when Tmx was not injected () Lmna+Tmx mice developed kyphosis (arrow head) by 21 days after injection. () LaminA/C protein, detected by immunofluorescence, were present in control (i, iii), but reduced/absent in cardiomyocyte (CM) nuclei in both isolated CMs (ii second panel) and heart sections (iv) (white arrowheads) with CM nuclei being detected by PCM-1 staining, 21 days after Tmx. () LaminA/C levels were quantified by Western analysis of whole heart lysates 21 days after injection. A significant reduction (***P=<0.0001; T-test) in A-type Lamin protein was detected, although Lamin C levels were not reduced as much in the Lmna+Tmx mice compared to Lmna+CTL. () Quantitative analysis was performed at 21 days post Tmx. The presence of the LoxP sites in the WT-Lmna gene (Lmna) results in a reduction in Lmna transcript levels compared to Lmnalevels, although this had no overt effect on longevity or growth/viability.
FIGS. 28A-28D
FIG. 28A
FIG. 28B
FIG. 28C
FIG. 28D
Flx/Flxmcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flxmcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
28
show echocardiograms, heart function and histology of Lmna+Tmx mice. () Lmna+Tmx mice show reduced cardiac contractile function. () Lmnahearts show reduced EF % and FS %, and increased LVID (***P=<0.0001, **P=0.0010 Two way ANOVA). () Histological analysis of the hearts revealed increased infiltration of nucleated cells and intercellular spaces in Lmnahearts (i and ii). Significantly fewer viable (brick-like) CMs were isolated from Lmnahearts compared to Lmnacontrols (iii). With higher magnification, the isolated cardiomyocytes from Lmnahearts contained large intracellular vacuoles (arrow head, iv). () The left ventricular lumen in Lmnahearts was enlarged (i) together with increased fibrosis (ii) (*P=0.0007 visible as lighter grey areas in the D ii, middle panels and iv left panel) and apoptotic nuclei revealed by TUNEL staining (*P=0.0220; One way ANOVA) (iii and iv right panel). All samples and analyses were performed on hearts 21 days post Tmx injection.
FIGS. 29A-29D
FIG. 29A
FIG. 29B
FIG. 29C
FIG. 29C
FIG. 29D
Flx/Flx:mcm
Flx/Flx:mcm
+/+
Flx/Flx:mcm
−/−
Flx/Flxmcm
+/+
Flx/Flx:mcm
+/+
Flx/Flx:mcm
−/−
Flx/Flx:mcm
+/+
Flx/Flxmcm
−/−
Flx/Flx:mcm
+/+
Flx/Flx:mcm
+/+
Flx/Flx:mcm
+/+
Flx/Flx:mcm
−/−
Flx/Flx:mcm
−/−
Flx/Flx:mcm
+/+
Flx/Flx:mcm
−/−
Flx/Flx:mcm
+/+
Flx/Flx:mcm
−/−
Flx/Flx:mcm
−/−
Flx/Flx:mcm
+/+
shows changes in nuclear morphologies and heart structure with and without Sun1 in the Lmnaafter Tmx injection. () CM nuclei with reduced or absent Lamin A/C expression are indicated by white arrow heads (1, 3). CM nuclei (2, 4) with normal Lamin A/C levels are indicated by grey arrowheads. LMNA protein levels, measured by both fluorescence intensity (5) and Western blot (6), were significantly reduced in LmnaSun1+Tmx (***P=0.0009; T-test) and LmnaSun1+Tmx (*P=0.0359; T-test) compared to LmnaSun1 controls (lower graph, 6) (). Left ventricular (LV) enlargement was apparent in the LmnaSun1+Tmx hearts (panel 1) but not in the LV of the LmnaSun1+Tmx hearts (panel 2). The LmnaSun1+Tmx mice had significantly increased fibrosis (panel 3, fibrosis in grey) compared to controls, but there was no significant increase in fibrosis in the LmnaSun1+Tmx hearts (panel 3) compared to controls (panel 4, quantified in panel 5, **P=0.0001; One way ANOVA). Cardiac papillary muscle active force measurements were significantly reduced from the LmnaSun1+Tmx mice compared to LmnaSun 1 controls (**P=0.0047; T-test) and LmnaSun1+Tmx (*P=0.0113; T-test) (panel 6). () CM nuclear morphologies were significantly altered in LmnaSun1+Tmx mice (Panel 1, solid arrow heads). In the absence of TMX, control heart sections (CTL, panel 2) display few nuclear abnormalities. In the absence of Sun1, LmnaSun1+Tmx cardiomyocytes showed no nuclear abnormalities (Panels 3 and 4). In summary panel 5 reveals that, 70% of CM in LmnaSun1+Tmx mice had NE ruptures/distortions or misshapen nuclei compared to less than 1% of CM nuclei in LmnaSun1+Tmx mice. () Echo analyses on TMX-treated and control mice were performed following Tmx induction. Echocardiograms (ECGs) performed at 28 days after Tmx injection on 3-5 month old mice (panel 1). ECGs performed before and after Cre induction revealed a progressive worsening of cardiac contractility in LmnaSun1+Tmx mice (solid black line) compared to LmnaSun1+Tmx mice (panels 2-4). The loss of SUN1 preserved EF (panel 2), FS (panel 3) and Global Longitudinal Strain (GLS, panel 4) in LmnaSun1+Tmx mice compared to LmnaSun1+Tmx mice.
FIGS. 30A-30B
FIG. 30A
FIG. 30B
N195K/Flx:mcm
−/−
N195K/Flx:mcm
+/+
N195K/−:mcm
+/+
N195K/−:mcm
+/+
N195K/−:mcm
−/−
N195K/Flx:mcm
−/−
N195K/Flx:mcm
+/+
show Kaplan Meier graph and heart function effects of deletion of SUN1 on cardiac pathology induced by a missense mutation in the Lmna gene (N195K). () The absence of Sun1 significantly increases the lifespan of LmnaSun1+Tmx mice compared to LmnaSun1+Tmx mice (*P=0.0101; Log-rank test). Mice with only one copy of the N195K mutation (LmnaSun1+Tmx) had an average lifespan of 47 days, approximately half the lifespan of mice homozygous i.e. with two copies of the N195K allele. () Echocardiograms (ECGs) performed before and after Cre induction revealed progressive worsening of cardiac contractility in LmnaSun1+Tmx mice compared to LmnaSun1+Tmx mice over time. ECGs images were recorded at 28 days after Tmx injection (left-hand side panels). The loss of SUN1 preserved EF, FS and GLS in LmnaSun1+Tmx mice compared to LmnaSun1+Tmx mice (right-hand side bottom 3 panels).
FIGS. 31A-31G
FIG. 31A
FIG. 31B
FIG. 31C
FIG. 31D
FIG. 31E
FIG. 20
FIG. 31F
FIG. 31G
Flxx/Flx:mcm
Flxx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
Flx/Flx:mcm
show Lmna+Tmx mice expressing an AAV transduced DNSun1 exhibit improved cardiac function and increased longevity. () Protocol for AAV-mediated transduction of the DN-Sun1 miniprotein into Lmna+Tmx mice. A single Tmx (IP) injection is given at D14 postnatally to induce Lmna deletion. AAV9-DNSun1 or AAV9-GFP viral particles are then injected into the chest cavity on D15 postnatally. The experimental endpoint was set at 100 days after Tmx. () The DNSun1 miniprotein competes with endogenous Sun1 for binding to the KASH domain of the Nesprins (in CMs this is Nesprin1). The miniprotein competes with endogenous SUN1 in binding to the KASH domain of the Nesprins. As the DNSun1 miniprotein is not anchored in the INM this effectively disconnects the endogenous SUN proteins from binding to the KASH domains so breaking the LINC. () The presence of the recombined Lmna gene following Tmx injection was confirmed by PCR of the heart tissues (upper panel). Robust expression of both AAV9-DNSun1 and AAV9-GFP protein (Dosage: 5×10∧10 vg/g of mouse) was detected in extracts from whole hearts 99 days post AAV injection (lower panel). () CMs derived from human iPS stem cells were transduced with the DNSun1 using AW-DJ as the vector. In CMs expressing high levels of DNSun1, indicated by grey arrows, Nesprin1 localization to the NE Is reduced or absent. Nesprin localization to the NE is maintained in CMs either not expressing the AW-DJ-DNSun1 or when expressed at lower levels (white arrow heads). () The Lmna+Tmx+AAV9-GFP mice lived for an average of 34.5 d after Tmx induction, whereas Lmna+Tmx mice injected with AA9-DNSun1 (5×10∧10 vg/g/mouse) lived significantly longer (**P=0.0038; Log-rank test) to at least 100 D post Tmx, after which the mice were sacrificed for analysis. This set of data was derived from that shown in , adjusted by removing mice that were female and those with a different dose of virus. Fig. E(i) represents male mice and Fig. E(ii) represents female mice. () At 35 d after Tmx, extensive fibrosis (blue in original image, grey here) and ventricular enlargement was detected in Lmna+Tmx+AAV9-GFP hearts compared to Lmna+Tmx+AAV9-DNSun1 hearts. () ECG analysis confirmed Lmna+Tmx+AAV9-DNSun1 hearts had better cardiac function compared to the Lmna+Tmx+AAV9-GFP hearts at 35 d days after Tmx injection.
FIGS. 32A-32D
FIG. 32A
FIG. 32B
FIGS. 32C
shows models of how breaking the LINC by disrupting Sun1 protects cardiomyocytes from contraction induced stress () Cardiomyocyte nuclei expressing LmnaA/C, are able to withstand mechanical stress and tension forces transmitted via the LINC complex from the cytoplasm to the NE. () The loss of or introduction of a mutation within the Lmna gene results in loss/or incorrect assembly of the nuclear lamina, which weakens the Lamina/NE. The weakened nuclei are damaged due to the tension/stress forces exerted via the LINC complex from the contractile sarcomeres of the cardiomyocytes. (, D). In the absence of SUN1 or by disrupting its binding to the KASH domains by expression of DNSun1, the untethered LINC complexes exert less tensional force on the cardiomyocyte nuclei, enabling survival of the Lmna mutant cardiomyocytes.
FIG. 33
Flx/Flx
Flx
Differentiation
: shows the structure of the Lmnaconditional allele. Primer locations for genotyping the Lmna gene both before and after Cre recombination are indicated for the Lmnaallele (Flox), the Lmna deleted allele (A) and the wildtype alele [A. S. Wang, et al., 89: 11-21 (2015)].
FIG. 34
J Cell Biol.
shows a diagram of the recombinant AAV9-DNSun1 and AAV9-GFP miniproteins. The DN-Sun1 includes the Sun domain, an HA tag, a Signal Sequence (SS, for targeting the protein to the ER), and the KDEL (ER retention signal) [M. Crisp et al., 172: 41-53 (2006)]. The AAV9-GFP includes the SS and KDEL sequences. GFP was used as a control in place of the Sun1L-KDEL.
FIG. 35
Flx/Flx:mcm
Flx/Flx:mcm
shows photomicrographs of cardiomyocyte specific expression of Cre recombinase after Tmx injection. The Lmnamice were crossed with the mT/mG (JAX: Gt(ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo/J) reporter mice. In the absence of Cre, RFP is expressed. When Cre is induced, GFP is expressed. Only CMs of Lmnamice express GFP upon TMX injection. Heart tissues were analyzed 7 days after Tmx injection.
FIG. 36
Δ/Δ
−/−
shows that loss of Sun2 does not rescue loss of Lmna. Loss of Sun2 does not extend the lifespan of Lmna Sun2 mice.
FIGS. 37A-37E
FIG. 37A
FIG. 37B
FIG. 37C
FIG. 37D
FIG. 37E
Flx/Flxmcm
+/+
Flx/Flxmcm
−/−
Flx/Flx:mcm
+/+
Flx/Flx:mcm
−/−
Flx/Flx:mcm
+/+
Flx/Flx:mcm
−/−
show the phenotypes of LmnaSun1 and LmnaSun1 hearts at 12-14 months after Tmx injection. () Histological analysis of the aged LmnaSun1 hearts, 12-14 months after the Tmx injection, revealing no significant morphological changes e.g. LV enlargement or () in fibrosis compared to the controls. () PCR analysis confirmed the sustained deletion of Lmna gene. () Protein quantification revealed a significant reduction of LMNA levels in LmnaSun1+Tmx hearts at 14 months after TMX. () Echocardiograms (left-hand side panel) from the aged mice showed reduced EF and FS (right-hand side panels) in both LmnaSun1+CTL and LmnaSun1+Tmx aged mice.
FIG. 38
Flx/Flx:mcm
shows the rescue by AAV9-DNSun1 depends on the dosage of viral particles injected. Lifespan of Lmna+TMX mice depends of the dosage of AAV9-DNSun1 with, with a lower concentrations resulting in shorter lifespans. Each dot represents a mouse, horizontal lines indicate mean.
FIGS. 39A-39C
FIG. 39A
FIG. 39B
FIG. 39C
show levels of LamiA/C following Tmx induction and expression of AAV-expressed proteins. () LaminA/C levels were significantly reduced following Tmx induction, and the presence of either AAV9-DNSun1 or AAV9-GFP protein did not alter LMNA protein levels (Quantification of LaminA/C immunofluorescence intensity). The amount of LaminA/C, DNSun1 and GFP protein in whole hearts were also quantified by Western analysis (lower 3 graphs). (Analysis performed 35 days after Tmx). () The expression of both DNSun1 and GFP proteins were dependent on the concentration of viral particles injected. () Immunofluorescence revealed the majority of CMs were successfully infected and expressed GFP with 5×10∧10 vg/g of AAV9-GFP (left image) compared to infection with a 10-fold lower (5×10∧9 AAV9-GFP, right image) concentration of viral particles.
FIGS. 40A-40C
FIG. 40A
FIG. 40B
FIG. 40C
show CRISPR targeting of Sun1 SUN domain results in loss of Sun1 protein. (A, B) Clustal alignment of Sun1 DNA () and amino acid () sequence (SEQ ID Nos: 69 and 72, respectively) adjacent to CRISPR-induced mutation in wildtype Sun1, Sun1 with 4 bp insertion (Sun1_plus4; SEQ ID NOs: 70 and 73, respectively) and Sun1 with 7 bp deletion (Sun1_del7; SEQ ID NOs: 71 and 74, respectively). Numbering is of Sun1 coding sequence (A) and protein sequence (B). Bold letters in (B) indicate SUN domain. () Immunofluorescence staining of mouse adult fibroblasts derived from wildtype and Sun1 mutant mice. Sun1 expression is lost in mutant mice, but Sun2 and Nesprin-1 expression is similar in all 3 genotypes. Scale bar=10 μm.
FIG. 41A-D
shows CRISPR targeting of Syne1 C-terminus results in expression of a mutant Nesprin-1 protein. (A, B) Clustal alignment of wildtype Nesprin-1 DNA (SEQ ID NO; 75) and Nesprin-1C′TΔ8 (Nesprn1_CTdel8) (SEQ ID NO: 76) (A) and amino acid sequence adjacent to CRISPR-induced mutation in wildtype Nesprin-1 (SEQ ID NO: 77) and Nesprin-1C′TΔ8 (Nesprin1_CTdel8) (SEQ ID NO: 78) (B). TGA in bold indicates stop codon of Syne1/Nesprin-1 gene. (C, D) Immunoblots of Nesprin-1 from Syne1/Nesprin-1 wildtype and Syne1/Nesprin-1C′TΔ8 mutant heart and muscle tissue.
FIG. 42A-B
are photomicrographs showing CRISPR-Induced Syne1 mutation results in mislocalized, “KASH-less” Nesprin-1 protein. Immunofluorescence staining of mouse adult fibroblasts (A) and primary myotubes (B) derived from widtype (WT) and Syne1C′TΔ8 mutant mice. Nesprin-1 is mislocalized from the nuclear envelope in the mutant samples. Merged images shows Nesprin-1 and DNA staining. Scale bar=10 μm.
FIG. 43A-C
are photomicrographs showing Syne1 mutation does not disrupt localization of certain nuclear envelope proteins. (A-C) Immunofluorescence staining of mouse primary myotubes derived from wildtype (WT) and Syne1C′TΔ8 mutant mice. Sun1 (A), Sun2 and emerin (B) and lamin A/C (C) localize normally to the nuclear envelope. Merged images show protein and DNA staining. Arrows indicate examples of normally localized nuclear envelope proteins. Scale bar=10 μm.
FIG. 44A-C
are photomicrographs showing Syne1 mutation disrupts localization of nuclear-envelope-localized centrosomal proteins. (A-C) Immunofluorescence staining of mouse primary myotubes derived from wildtype (WT) and Syne1C′TΔ8 mutant mice. Pcm1, Pericentrin (Pcnt), and Akap450, which normally localize to the nuclear envelope in myotubes, are displaced from the nuclear envelope in Syne1C′TΔ8 mutant myotubes. MF20 is an antibody for myosin heavy chain, a myotube marker. Merged images show protein and DNA staining. Arrows indicate typical nuclear envelope staining for these centrosomal proteins. Scale bar=10 μm.
FIG. 45A-C
shows Syne1 mutation does not affect mouse phenotype. (A-B) Representative images of 12-week-old male (A) and female (B) mice. (C) Bodyweight of male and female, wildtype (WT) and Syne1C′TΔ8 mutant, mice over 6 weeks.
FIGS. 46A-46C
FIG. 46A
FIG. 46B
FIG. 46C
C′TΔ8/C′TΔ8
−/−
shows Syne2 constructs and Syne1/Syne2 double mutant mice experience perinatal lethality. () Design of IRES-βgal PGK-Neo targeting construct for generating Syne2 mutation. () Immunofluorescence staining of mouse adult fibroblasts derived from wildtype (WT) and Syne2 mutant mice showing loss of Nesprin-2. () Images of newborn pups. Top row are of mice with at least 1 wildtype Syne1 or Syne2 allele that appear a healthy pink. Bottom row shows cyanotic double mutant Syne1:Syne2 pups which appear blue and die at birth.
FIG. 47
Δ/Δ
is a Kaplan Meier graph showing a Syne2 mutation does not ameliorate Lmna pathology. Kaplan-Meier survival curve showing that regardless of their Syne2 mutation status (wildtype, heterozygous or mutant), Lmna mice die within 3 weeks of birth. | |
Home News Jam Master Jay: List Of Events Surrounding The Murder And The Key Players..
Jam Master Jay: List Of Events Surrounding The Murder And The Key Players..
Five years after the shocking murder of legendary DJ Jam Master Jay of Run-DMC, theories and emotions continue to swirl. They’ve heated up in recent months and culminate this week, as MTV News presents Randy Allen, one of the witnesses to the murder, speaking with Marvin Thompson, Jay’s brother, about the tragedy for the first time in five years.
As the first installment of a three-part series rolling out this week culminating in the first conversation between Jay’s brother and Randy Allen, the DJ’s former business partner, since the murder, MTV News has compiled a timeline of the events surrounding the murder.
October 30, 2002, 6:00 p.m.: Jay arrives at his 24/7 Studio in Queens, New York, to work on some material for Rusty Waters, one of the acts signed to his label. Waiting for things to get started, he sits on a couch in the studio’s lounge and plays a few games of “Madden 2002″ with his friend Uriel “Tony” Rincon, 25. According to Rincon, Jay brings out a pistol and places it on the couch next to them, which makes the studio’s receptionist and Jay’s assistant, Lydia High, nervous. She asks him to put it away when she comes in a short time later to discuss Jay’s schedule. Randy Allen, a member of Rusty Waters and a longtime friend of Jay’s, arrives at the studio soon after, along with a friend known as Mike B., and the pair go into another room to listen to demo tapes of Hot Ta Def, an aspiring artist on Jay’s label.
October 30, 2002, 7:30 p.m.: According to Rincon, his cell phone rings around this time, and as he reaches down to answer it, he hears footsteps. It is unknown how many people are involved, but the assailant or assailants walk by High’s desk, telling her to get on the ground, and then greet Jay. The DJ reportedly embraces and/or slaps hands with his shooter, who appears to have been buzzed into the studio by High, indicating that the DJ knows his assailant or assailants. According to Rincon, the shooter opens fire, striking him in the leg and Jay, 37, fatally in the back of the head. Though Allen, Mike B., Rincon and High are all at the studio that night, reportedly none of them witness the actual shooting or see who pulled the trigger. Someone from the control room where Allen and Mike B. are working reportedly follows the gunmen as they leave and fires a weapon at them.
November 2002: Theories begin to emerge about the possible motives for the murder, including a debt owed to an old neighborhood acquaintance, revenge by disgruntled rappers with whom Jay had worked and who may have killed him over a disputed music publishing advance, or even a possible connection to 50 Cent, who had been a protege of Jay’s.
November 10, 2002: The New York Post reports that Ronald “Tinard” Washington, an associate of Jay’s, may have served as a lookout while the DJ was shot. Washington is believed to have tipped off the killers when Jay arrived at his 24/7 Studio and then waited outside while the perpetrators committed the crime. The man believed to be an accomplice to Jay’s murder is thought to be hiding out in Washington, D.C. He was only identified as “a convicted drug dealer.” The paper also says that Jam Master Jay may have been killed for a dispute over money, and names Curtis Scoon who had allegedly been feuding with Jay as the prime suspect in the investigation and the man wanted by police for questioning. Among the theories is that Jay owed a debt to Scoon and was killed following an argument over the money.
December 12, 2002: Jay’s longtime friend, Randy Allen, who was present in the studio the night of the murder, denies reports that he was involved in the slaying and had set up Jay in an attempt to cash in on a $500,000 life insurance policy. “You can’t believe everything you read. That’s a cruel thing to do to Jam Master Jay,” Allen later told MTV News.
June 19, 2003: An investigator tells MTV News that police are again looking into reports that Allen may have been involved in the murder. The news comes days after Jay’s mother, Connie Mizell, told New York’s Daily News that she was upset she had not heard from Allen since the DJ’s slaying and about reports he wasn’t cooperating with police. “That hurts me more than words will ever say,” she told the paper. “All of these years, [Jay] and Randy have been friends. We were all as close as close can be, and I haven’t seen Randy since my Jason was killed. You’re his friend for 20 years and you don’t want to talk to the police about what happened? You don’t come to my house after he died? You want to say you don’t know anything?” Police dismissed, however, another report that same week that the murder may have resulted from a deadly love triangle between Jay, his wife, Terri Corley-Mizell, and Kenneth “Supreme” McGriff, an alleged neighborhood drug kingpin. | http://www.hiphoplead.com/news/jam-master-jay-list-of-events-surrounding-the-murder-and-the-key-players/ |
The Arizona Supreme Court recently ruled on the common law precedent of the original tortfeasor rule, which allows a person who is civilly liable to another for some injury may also be civilly liable for the negligence, mistake or lack of skill on the part of a doctor or surgeon who treats the injured person for that injury.
What is often considered is whether aggravation of the original injury or subsequent additional injury due to poor medical or surgical treatment was a natural and probable consequence of the original injury or a direct result of the original tortfeasor’s actions.
In Florida, this issue was weighed by the Florida Supreme Court in Stuart v. Hertz Corp., back in 1977. In that case, our highest court ruled that a rental car company whose vehicle injured a woman in an accident could be liable for injuries she sustained from medical negligence while she was being treated for her original injuries. The court further ruled the rental car company couldn’t introduce the doctor into the case and make him pay for his portion of the damages. (The federal Graves Amendment has since barred individuals from taking action against rental car companies for the negligence of their driving customers, though that is beside the point here).
Per the comparative fault regime, the court concluded the original tortfeasor rule does not stop the defendant from alleging or proving that a non-party physician who treated the plaintiff for injuries sustained as a result of defendant’s negligence should be held to account. However, the original defendant can by liable for injuries sustained in the course of treatment that was reasonable for the original injuries, even if those injuries were negligently caused by the doctor, so long as they were foreseeable.
Essentially, as our West Palm Beach car accident attorneys can explain, it’s not automatic that the original defendant will be held liable for the negligent actions of a doctor, but it will depend on the court’s assessment of allocation of fault between parties and non-parties.
In the Cramer case, one woman rear-ended another in which plaintiff was a passenger. Plaintiff suffered numerous headaches and lower back injuries, for which she sought chiropractic treatment. It was later revealed that her persistent pain was attributable to several disc protrusions of her lumbar spine.
Eight months later, her doctor recommended surgery. However, that procedure didn’t cure her condition and in fact may have made it worse.
After the unsuccessful surgery, plaintiff filed injury action against defendant driver. At request of defendant, plaintiff underwent an independent medical exam and concluded plaintiff’s injuries were not the result of the car accident, but rather due to the unsuccessful surgery.
Based on that, defendant filed notice of naming the surgeon as a non-party at-fault. Plaintiff sought a motion to strike, arguing defendant, as the original tortfeasor, was liable for the foreseeable risks of her tort. Trial court agreed and granted motion to strike.
The trial court reversed. Although it is possible, the court ruled, for the original defendant to be held liable in some instances, for medical negligence stemming from treatment of those original injuries, the original tortfeasor rule doesn’t preclude a defendant from asserting – or a court from finding – that a non-party physician is also liable. | https://www.south-florida-injury-lawyer-blog.com/cramer-v-hon-starr-original-tortfeasor-rule-tested/ |
With so many organizations today embracing all the latest technologies out there, having a solid IT infrastructure matters more than ever. Not only is downtime an inconvenience to organizations, but it can easily drive them out of business altogether. According to a study by Gartner, the average cost of IT downtime is $5,600 per minute. This is why organizations need system administrators to keep all their business applications running and up to date.
System Administrator - Overview
A system administrator is responsible for maintaining the entire IT infrastructure of an organization. Their work ranges from maintaining servers to updating software and everything else in between. They are the first point-of-contact for any technical problems that employees face. When no one in an organization faces any issue while doing their job, this means that systems administrators have done their job well.
System Administrator - Roles and Responsibilities
The roles and responsibilities of a system administrator can vary widely from one organization to another. Here are the four types of system administrators based on their roles and responsibilities:
Network Administrators
Network administrators manage the entire network infrastructure of an organization. They design and install computer systems, routers, switches, local area networks (LAN), wide area networks (WAN), and intranet systems. They also monitor the systems, provide maintenance and troubleshoot any problems when they arise.
Database Administrators
Database administrators (DBA) set up and maintain databases used in an organization. They may also be required to integrate data from an old database into a new one or even create a database from scratch. In large organizations, there are specialized DBAs who are only responsible for managing databases. In smaller organizations, the roles of DBAs and server administrators can overlap.
Server/Web Administrators
Server or web administrators specialize in maintaining servers, web services and operating systems of the servers. They monitor the speed of the internet to make sure that everything runs smoothly. They also analyze a website’s traffic patterns and implement changes based on user feedback.
Security Systems Administrators
Security systems administrators monitor and maintain the security systems of an organization. They develop organizational security procedures and also run regular data checkups - setting up, deleting and maintaining user accounts.
In large organizations, all these roles may all be separate positions within one department. In smaller organizations, they may be shared by a few system administrators, or even one single person.
System Administrator - Skills
Since maintaining an entire IT infrastructure is not an easy feat, system administrators need to have a mixture of hard and soft skills. Here are a few skills that companies look for in system administrators.
- Knowledge of different operating systems (Microsoft, Unix, Linux, etc.)
- Hardware knowledge
- Attention to detail
- Problem solving mindset
- Critical thinking
The most important skill for a system administrator is the ability to solve problems under pressure. Since every minute is critical for the success of a business, a system administrator should be able to quickly diagnose what is wrong and how best to fix it.
System Administrator - Educational Qualification
The basic requirement for a system administrator is a bachelor’s degree in Computer Science, IT, IT systems management, or any other related field. However, you can also opt for certification programs with a core emphasis on hardware, computer networks, and system administration. An updated professional certification is the quickest tool that employers can use to check your skills.
Here are some popular certifications that will help you gain new skills, earn more money, and stay ahead of the competition:
- Microsoft Certified Solutions Expert (MCSE)
- VMware Certified Professional 6-Data Center Virtualization
- Oracle Linux System Administrator (Oracle)
- ITIL® Certification
- Red Hat Certified Engineer (RHCE)
System Administrator - Job Description
The career outlook for system administrators continues to grow along with the changes in technology. Hence, they are required to stay aware of all the latest trends in the tech space. These days, most companies require certification or recertification to ensure that their employees are not falling behind. According to the US Bureau of Labor Statistics, systems administrator roles are projected to grow 4 percent from 2019 to 2029.
Here are some common requirements that you would find in a system administrator job description:
- Install and configure software, hardware and networks
- Ensure security and efficiency of IT infrastructure
- Monitor system performance and troubleshoot issues
- Identify system requirements and install upgrades
- Maintains, secure and upgrade a web system
- Create a backup and safeguard the data
- Perform account setup for new and old employees
- Track emerging technologies and implement them in the organization
System administrator - Salary Trends
A system administrator’s salary can depend on the location, experience and the type of systems they manage. Here is a list of system administrators’ salaries around the world.
- Annual average system administrator salary in India - ₹692,255
- Annual average system administrator salary in US - $82,925
- Annual average system administrator salary in UK - £25,634
- Annual average system administrator salary in Canada - CA$59,961
- Annual average system administrator salary in Australia - AU$68,685
- Annual average system administrator salary in Singapore - S$44641
- Annual average system administrator salary in UAE - AED 84,014
- Annual average system administrator salary in Saudi Arabia - SAR 92,37
Are you looking forward to becoming an ITIL expert? Check out the ITIL Foundation Certification Course and get certified.
Here’s Your Next Step to Success
Systems administration is a career path where experience significantly outweighs a college degree. They need to have proper training and experience before taking over an IT infrastructure completely. ITIL certified professionals can earn up to 40-percent more than their non-certified peers. If you want to understand the concept of ITIL frameworks and enhance the quality of IT Service Management, Simplilearn’s ITIL® 4 Foundation Certification Training Course is the perfect place to get started. This ITIL V4 certification course will help you understand the concepts, languages, best practices, and processes utilized in the ITIL 4 lifecycle. Whether you are just getting started or are a seasoned IT professional, you need to update, renew, and even reactivate your certifications to ensure that you are well equipped with the latest skills in information technology. Get trained and certified today with Simplilearn to help you get ahead in your career. | https://www.simplilearn.com/systems-administrator-article?source=frs_author_page |
Is minilaparoscopic inguinal hernia repair feasible? A preliminary experience.
Laparoscopy has rapidly emerged as the preferred surgical approach to a number of different diseases because it allows for a correct diagnosis and proper treatment. It seems to be moving toward the use of mini-instruments (5 mm or less in diameter). The aim of this paper is to illustrate retrospectively the results of an initial experience of minilaparoscopic transabdominal preperitoneal (miniTAPP) repair of groin hernia defects performed at two institutions. Between February 2000 and December 2003, a total of 303 patients (mean age, 45 years) underwent a miniTAPP procedure: 213 patients (70.2%) were operated on bilaterally and 90 (28.7%) for a unilateral defect, with a total of 516 hernia defects repaired. The primary endpoint was the feasibility rate for miniTAPP. The secondary endpoint was the incidence of mini-TAPP-related complications. No conversions to laparoscopy or an anterior open approach were required. There were no major complications, while minor complications ranged as high as 0.3%. While limited by its retrospective design, the present study indicates that the minilaparoscopic approach to groin hernia repair is safe and effective, making miniTAPP a challenging alternative to laparoscopy in the approach to groin hernia repair.
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Warblers are what they eat: Distinctive morphologies influence dietary variation among coexisting species
Thinking about dietary specialization in birds may elicit images of raptors carrying animals in their sharp, curved beaks or shorebirds using their long, slender bills to probe for invertebrates. However, dietary adaptations can be subtler. The saying “you are what you eat” has long been assumed to apply to many animal species, and birds in particular, in the sense that form, or body shape, is associated with function, or the ability to capture and eat particular foods. We delved deeper into this pattern, finding that not only are the two closely linked in New World wood warblers (Parulidae), but also the chief external morphological variations in these birds are directly linked to diet in multiple ways. Specifically, the very morphological differences that distinguish coexisting warbler species and resulted from these birds’ adaptive radiation correlate with diet.
New World warblers generally have small bodies and short beaks, and many species are primarily insectivorous and sport brightly colored plumage. Aside from their often bright and distinctive plumages, the numerous parulid warblers appear surprisingly similar at first glance, but their subtle differences in external morphology are more important than previously recognized. For example, American Redstarts have flat beaks, long and pointed wings, and long rictal bristles that equip them for aerial flycatching. Meanwhile, Black-and-white Warblers have deep, curved claws that facilitate tree-creeping movements to search for insects in bark.
Our study used diverse measurements of beak, tarsi, tail, and wings based on museum study skins to reveal little morphological overlap among warbler species coexisting in both breeding and wintering communities. Our results also supported the well-established relationship between form and function related to feeding, with larger warblers eating larger beetles and orthopterans. A more novel result was that the subtle morphological differences among species were associated with key differences in diet: Warbler species with aerial foraging morphologies consumed proportionately more winged insects.
Diet data are scarce in ornithological studies, which is likely in part because it is easy to use foraging behavior or morphology as surrogates for diet. However, we obtained detailed, quantitative diet data from warbler gut contents by identifying arthropods and arthropod fragments, including heads or partial heads, elytra, mandibles, legs, and wings to either order or family. Our methods bridged ornithological and entomological knowledge to elucidate patterns relevant to interspecific competition and adaptive radiation, i.e., to the ecology and evolution of these birds.
Our results indicate that these migratory warblers’ diverse diets are likely important to their coexistence in both breeding and wintering communities. Moreover, our findings suggest that natural selection relevant to foraging efficiency has likely contributed to the morphological variation that characterizes warblers’ adaptive radiation, which is one of the most dramatic in North American birds. A key takeaway is that the primary morphological differences in these warblers directly influence their diets, or more simply, they are what they eat.
This research was recently published in the Journal of Field Ornithology:
Rosamond, K.M., C.M. Kent, S.C. Hunter, and T.W. Sherry. 2020. Morphological traits influence prey selection by coexisting species of New World warblers (Parulidae). Journal of Field Ornithology https://doi.org/10.1111/jofo.12352.
Guest post by the authors of the new paper in JFO. | https://afonet.org/warblers-are-what-they-eat-distinctive-morphologies-influence-dietary-variation-among-coexisting-species/ |
#reddirect density dependence
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Indian Govt Plans to Stop Fake News on Social Media Sites
India’s Information Technology Act (IT Act), 2000 was enacted with a view to give a fillip to electronic transactions, to provide legal recognition for e-commerce and e-transactions, to facilitate e-governance, to prevent computer based crimes, and ensure security practices and procedures. The Act came into force on 17th October, 2000.
Section 79 of the IT Act elaborates on the exemption from liabilities of intermediaries in certain cases. Section 79(2)(c) mentions that intermediaries must observe due diligence while discharging their duties, and also observe such other guidelines as prescribed by the Central Government. Accordingly, the Information Technology (Intermediaries Guidelines) Rules, 2011 were notified in April 2011.
[ Indian Govt May Regulate Social Media Before 2019 Lok Sabha Election ]
The Indian Government said Monday it is committed to freedom of speech and expression and privacy of its citizens as enshrined in the Constitution of India, adding that the Government does not regulate content appearing on social networking platforms.
These social networking platforms, though, are required to follow due diligence as provided in section 79 of the Information Technology Act 2000 and the Rules notified therein, subject to the import of Article 19(2) of the Constitution and ensure that these platforms are not used to commit and provoke terrorism, extremism, violence, and crime.
Government says the instances of misuse of social media by criminals and anti-national elements have brought new challenges to the law enforcement agencies. These include inducement for recruitment of terrorists, circulation of obscene content, spread of disharmony, incitement of violence, public order, fake news, etc.
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According to the Government, a number of lynching incidents were reported in 2018 mostly alleged to be because of fake news / rumours being circulated through Whatsapp and other social media sites.
A calling attention motion on “Misuse of Socal Media Platforms and Spreading of Fake News” was admitted in the Parliament (Rajya Sabha) in 2018 (Monsoon Session).
India’s Minister for Electronics and IT, responding to the calling attention motion on 26/07/2018, made a detailed statement where he inter alia conveyed to the House the resolve of the Government to strengthen the legal framework and make the social media platforms accountable under the law.
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Subsequently, the Ministry of Electronics and Information Technology (MeitY) prepared the draft Information Technology (Intermediary Guidelines) Rules 2018 to replace the rules notified in 2011.
Presently consultation process is underway. The consultation process was initiated with Inter-Ministerial consultations and thereafter with other stakeholders,including major social media companies / messaging services platforms like Facebook, Google, Twitter, Yahoo, WhatsApp, and other associations.
The Government says it intends to consult all stakeholders in the process. Accordingly, MeitY has commenced public consultation on the draft rules with the timeline of submission of comments by 15th January, 2019.
Support RMN News Service for Independent Fearless Journalism
In today’s media world controlled by corporates and politicians, it is extremely difficult for independent editorial voices to survive. Raman Media Network (RMN) News Service has been maintaining editorial freedom and offering objective content for the past more than 12 years despite enormous pressures and extreme threats. In order to serve you fearlessly in this cut-throat world, RMN News Service urges you to support us financially with your donations. You may please click here and choose the amount that you want to donate. Thank You. Rakesh Raman, Editor, RMN News Service. | http://www.ramanmedianetwork.com/indian-govt-plans-to-stop-fake-news-on-social-media-sites/ |
The Constitutional controversy initiated by the Health Care Bill has brought the Commerce Clause in the Constitution back into public view. It simply states:
The Congress shall have Power … To regulate Commerce with foreign Nations, and. among the several States, and with the Indian Tribes;”- U.S. Constitution, Art. I, § 8, cl. 3
While those 16 words seems simple enough, the people we have elected to represent us seem to have a great deal of trouble understanding them. As far as our founders were concerned, this was a very practical clause that gave the central government the ability to keep trade flowing throughout the new nation. They wanted to guard against unrealistic regulations imposed by any state that could impair the flow of trade throughout the nation.
Alexander Hamilton explains the Commerce Clause
To explain the proposed Constitution in particular to the ratifying convention in New York, whose governor opposed it, Alexander Hamilton, John Jay and James Madison wrote a series of 85 articles that were published in New York City newspapers under the pseudonym of Publius. These essays became known as the Federalist Papers.
Federalist No. 11 by Alexander Hamilton explains the Commerce Clause in the Constitution. It is entitled: “The Utility of the Union in Respect to Commercial Relations and a Navy.” In this essay he stated:
An unrestrained intercourse between the States themselves will advance the trade of each by an interchange of their respective productions, not only for the supply of reciprocal wants at home, buy for the exportation to foreign markets…Commercial enterprise will have much greater scope from the diversity in the productions of different States. When the staple of one fails from a bad harvest or unproductive crop, it can call to its aid the staple of another.
Hamilton went on to explain that without a union between the States that trade would be less successful; would be fettered and interrupted and narrowed by many causes.
The 16 words of the clause and the intentions of the founders would hardly seem like an appropriate subject for a present-day article. The words in the clause are clear and the essay by Hamilton makes the intention of the clause abundantly clear. Unfortunately, it is an appropriate subject matter when you have a group of nine elite judges deciding whether or not the Commerce Clause can be used to justify Obamacare.
The Commerce Clause could fix Obamacare
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Here’s the question: Can the Commerce Clause force people to buy something?
Here’s the irony: The Commerce Clause can correct one of the problems with healthcare – the problem of not being able to buy something across state lines. More irony – that is not one of the objectives within Obamacare. Here in New Jersey where healthcare premiums soar to unprecedented heights, buying across state lines would provide competition between carriers and allow citizens to buy or not to buy the product of their choice. That choice doesn’t exist; healthcare as written does not address the issue as it could with proper reliance on the Commerce Clause; and the clause is being perverted to accommodate the man now occupying the White House’s personal agenda.
- Question: What will the United States Supreme Court decide?
- Answer: Whatever their ideology dictates.
Once again, monumental decisions that will affect the lives and prosperity of many of our citizens will not be rooted in the Rule of Law prescribed in our founding documents, nor will those decisions be based on the clear intentions of our founders. They will be based on political ideologies that have morphed this nation into something that would be unrecognizable to our founders. As a wise old friend often says,
The law means whatever the judges say it means when they say it.
Silly me; I always thought the law was based on our Constitution.
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CATEGORY:Constitution
Let’s see, all Congress had to do, then, was impose an additional tax that would be enough to pay for minimal health insurance for every citizen, and there would have been no Constitutional problem since Congress is empowered to levy taxes.
Except, the ACA passed by Congress requires that people buy insurance from private companies in an open, free market, or pay a penalty in the form of a tax surcharge. How anti-capitalistic can you get?
If no one has to carry insurance, then they can cross a state line, get injured, and get free emergency room care at a hospital in that state. They’re then free to return home, and while the bill collectors may hound them, the cost of care is now the problem of another state that has no local jurisdiction over the person who incurred the expense.
Since healthcare is something virtually guaranteed to be used by every citizen at some point in life, and since citizens are free to travel across state lines at any time in their lives, and since there is no ability to control when emergency care will be needed or the amount a given incident will cost, this seems entirely in line with the kind of activity within the scope of the Commerce Clause.
One could also reverse the entire argument and state that the ACA uses a tax-penalty mechanism to encourage an activity, and leaving the freedom to choose the activity or the tax surcharge up to each individual. Citizens are free to decide whether it makes more sense in each individual case to pay for free-market health insurance or a tax surcharge as they see fit. Congress has the Constitutional power to tax, so how is this illegal?
inwhile leaving the achoicerather than forcing the activity under some other form of pe
Sorry – the last line in my prior reply is stray text I forgot to delete.
Looks like that was the basic logic CJ Roberts used – Congress is within it’s Constitutional rights to define law using taxes as an incentive or disincentive for action.
Haven’t read through the ruling documents yet, but the comments from him that I’ve seen recapped also make an obvious point – the role of the Supreme Court is to interpret the laws passed, not to create law, and if you want to change the law, vote for members of Congress who’ll do that.
download identity 2003 Great text. I like your web design also. continue your good work.
Great text. I like your web design also. continue your good work.
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- This facility can be used by PAN applicants having a valid Digital Signature Certificate (DSC) issued to them by authorized Certifying Authority (CA) in India.
- PAN applicants should have scanned image of photograph, signatures and supporting documents i.e. Proof of Identity (POI) /Proof of Address (POA) /Proof of Date of Birth (PODB). The photo/signature and supporting documents should be scanned as per following specifications:
-
Sr.No Parameters Photograph Signature Supporting documents 1. Resolution (in DPI) 200 DPI 200 DPI 200 DPI 2. Type Color Color Black & White 3. File type JPEG JPEG PDF/A or JPEG 4. Size Max. 20 KB Max. 10 KB Max. 300KB/per page 5. Dimension 3.5X2.5 cms. 2X4.5 cms
Reduce Signature Size Online
What should be the size of the scanned photograph and the signature?
The candidate should scan his/her signature which has been put on white paper with black ink pen. Each of the scanned images of the photograph/signature should not exceed 40 KB in size (for photograph, the pixel size is 140 pixels height x110 pixels width and for sign it should be 110 pixels height x140 pixels width) and also to be ensured that the uploaded photograph/signature are proper.
If the Photograph is not of the proper pixel size given, then how to convert it?
Use any free image editing software such as MSPaint or Irfanview. To delete the unnecessary print area, use the CROP option after selecting the Image. To resize to proper pixel size use the resize option. Help for cropping / resizing can be seen here>>
. If the Photograph is not in the desired format, then how to convert it?If the image is in other formats such as .tiff,.bmp etc, then open the photo in MsPaint or Irfanview and click save as option to save it in the desired format (.jpg or .png).
How do I load my signatures/photograph?
The scanned photograph and signature in the format and the specification which has already been explained above is to be uploaded.
My photograph and signature as I loaded on the site are not appearing right. What should I do?
After uploaded the photograph and the signature a fresh page shows the preview of the uploaded images. If, the applicant is satisfied with the uploaded images he may click on Confirm Uploadbutton. In case the applicant is not satisfied with the uploaded images, he may click on the Reload Photograph or Reload Signature button. The reloaded photograph/ signature can again be viewed by clicking on the Image refresh button. The process can be repeated till satisfaction. The Candidates may please note that if the quality of photograph/signature is poor, the application is liable to be rejected After having clicked on the Confirm Upload button, the system will not allow any changes in the photograph/signature. However, if an applicant is not satisfied with the format/size/quality of the photograph or signature, the applicant may submit a fresh online application with the revised photograph and signature along with prescribed fee to be paid again. | https://customercarehotline.com/pan-card-photo-signature-resizer/ |
New Student Guide
Welcome Class of 2026!
We are excited to have you join our community, and this guide highlights key information and university requirements. Just follow the directions below to ensure your smooth transition to Georgetown University in Qatar (GU-Q). In the summer, new students will receive an invitation to enroll in Hoyas 101, a special orientation program to learn more about life, services and support at GU-Q. We look forward to seeing you in July!
Start Here!
Congratulations on your acceptance! To process your enrollment, you need to submit a few documents and applications. You will also need to activate your NetID and Georgetown email account.
Health & Wellness
Learn about immunization compliance, health insurance, and emergency contact information.
NSO & Convocation
Find out about the mandatory New Student Orientation (NSO) and the Convocation ceremony which officially makes you a GU-Q student.
International Students
Information about immigration, airport transportation, orientation, housing, and what to pack.
Program Information
View the calendar and available courses, and understand requirements including the core curriculum, pre-registration, and foreign language proficiency.
Academic Resources
Find out about academic accommodations and academic support services offered.
Useful Information
View other useful information regarding IT services, facilities in the GU-Q building, finances, and student employment.
Student Housing
Learn how to apply for housing and the housing dates!
Start Here!
Submit your enrollment deposit by May 1, 2022 to reserve your place in the incoming Fall 2022 Class.
For questions, please contact The Office of Admissions.
After you have submitted a deposit to secure your place in the incoming class, you will receive a letter from the Office of Admissions with information about your NetID, along with a temporary password. This step is very important to access all services.
Password management is critical for the security of your work and personal accounts. Check out the updated NetID password standard and videos and other information about how to navigate password security and management!: https://security.georgetown.edu/best-practices/password-management
Please change your temporary password by going to http://password.georgetown.edu/ within the next 48 hours. This site will enable you to securely enroll in Password Station to set responses to security questions, DUO two factor authentication and create a permanent password.
If you have any questions regarding your NetID password, please contact the GU-Q IT User Support at +974 4457 8130 and if you need assistance in resetting the password then please visit https://admissions.qatar.georgetown.edu/register/NewStudentPasswordReset2022.
Your university e-mail address is «NetID»@georgetown.edu. After setting a permanent NetID password, you can log into your Georgetown email at https://apps.georgetown.edu/.
Be sure to check your Georgetown University email account for all official Georgetown correspondence. Official University messages are sent only to your Georgetown email address. For information about using Google Apps, please go to https://uis.georgetown.edu/google-apps/.
Have Your Final Transcript Sent to GU-Q
Final transcripts must be sent to Georgetown University in Qatar as soon as they become available. Transcripts submitted by students are considered unofficial and will not be accepted. Official transcripts should be received directly from your school/counselor via email or online from the examination board (IB, Cambridge, EdExcel, College Board… etc).
If you have any questions or concerns regarding your high school transcripts, please contact the Office of Admissions.
Advanced Credit for Credit at GU-Q
If you have completed an advanced credit program, such as Advanced Placement, International Baccalaureate, or British A/AS-Levels, you might be eligible for university credits that will impact your fall class schedule. Report your scores to GU-Q Academics as soon as they are available. Unofficial scores can be reported as long as you follow up with the official scores when they are available.
Transfer Students
If you are a transfer student, you must provide an official copy of your final academic transcripts from previous academic institutions, as well as course syllabi, in order to evaluate the courses for transfer. It is important to receive transcripts early, as these can affect fall class registrations. Transcripts and syllabi can be submitted by email to GU-Q Advising.
You are required to submit a digital passport size photo for your student ID card. Your ID card will allow you to have access to the Georgetown building, as well as, access a variety of functions such as the student lounge, library and transportation services. Failure to provide a photo will delay the issuance of your card and may prevent you from accessing the Georgetown building and other university services.
Your photo needs to be in color, on a solid background and facing the camera. Photos with hats, sunglasses, as well as partial photos will be rejected.
► Please upload your digital photo via your Application Status Page.
Please follow this guide for your digital photo:
If you are a resident and have a Qatar ID, a copy must be submitted via your Application Status Page. If you are applying from outside Qatar and already have a Qatar ID, or have an old ID, please submit a copy.
Health and Wellness
All students are required, at the time of registration, to provide documentation of vaccination or immunity from Polio, Diphtheria, Tetanus, Hepatitis B, Measles, Mumps, Rubella, Varicella, and Meningitis. In addition, all students are required to have documentation of a Tuberculosis (TB) skin test in the last year and submit the Immunization History and Tuberculosis Screening form upon admission.
If you choose to waive the Meningitis vaccination, you should read the Meningitis Fact Sheet and complete and sign the Meningitis Vaccine Waiver.
The Immunization History Form and Tuberculosis Test Form must be completed by a health care physician. This process can take several weeks to complete. You will need to plan ahead to meet the deadline and avoid registration delays.
► Please upload your completed immunization documents via your Application Status Page. PLEASE DO NOT EMAIL YOUR COMPLETED FORM OR IMMUNIZATION DOCUMENTS TO US.
Deadline to submit your required immunizations is JUNE 15, 2022.
For more information on required immunizations, click here.
For inquiries, contact the Student Wellness & Counseling Center.
All enrolled international and local expatriate students residing in Qatar are required to carry a supplementary private health insurance; therefore, GU-Q has negotiated a comprehensive medical and travel policy designed specifically for GU-Q students.
All international and local expatriate students will be automatically enrolled and charged a premium fee for this insurance on their student billing accounts, unless they decline the coverage and provide documentation of adequate insurance coverage under another private policy.
If you have coverage by another private insurance that remains in effect throughout the academic year, and you wish to decline the student health insurance arranged by GU-Q, you must complete the Insurance Waiver Form and provide a copy of the front and back of your current private health insurance card. ► If you are eligible to opt out of enrollment in GU-Q insurance plan, please upload your completed Insurance Waiver Form along with a copy of the front and back of your current private health insurance card via your Application Status Page, should you choose to waive.
Students are responsible for understanding their insurance coverage and benefits and must carefully read the enrollment and benefit details, including the inclusions, exclusions, and limitations of their coverage.
Deadline to waive out of student insurance is JUNE 1, 2022.
For more information on Student Health Insurance and Medical and Travel Insurance Policy, click here.
For inquiries, contact the Student Wellness & Counseling Center.
You need to provide the following two pieces of information by logging into https://myaccess.georgetown.edu/ before you arrive. You will need your Net ID and password.
- Emergency contact information (Next of Kin tab). Please provide a name, relationship, address and mobile number for the person you want us to call in case of an emergency. You may add more than one person.
- Your mobile number (Addresses and Phone tab). We use your mobile number to reach you by SMS only when needed. For those who are coming from abroad, you can change the mobile/cell phone number later, once you get a Qatar SIM card. Use the format 974######## when entering your number.
Student Housing
This is a two-step process. First, Georgetown needs to verify that you are a student. Once this is done, students will shortly receive an email from StarRez. You should respond to the instructions sent by the StarRez system.
Information for students who apply for housing will automatically be provided to QF Housing and Residence Life (HRL). HRL will then send information to your Georgetown email with instructions for completing your housing application through their StarRez system.
Students who already live in Qatar are also eligible to apply for housing.
Steps to Apply
- Step 1: Please click on this Link in order to start the application process for QF Residence Halls.
- Step 2: Look for an email from Qatar Foundation / StarRez and follow the instructions.
Expected Assignment Timeline
By the end of June, Qatar Foundation will email applicants a link to the StarRez system. You must follow the instructions emailed to you in order to reserve a space. Note that you will not hear back from QF Housing Residence Life (HRL) about your room assignment until the beginning of July.
Students can check into QF Student Housing as early as July 15, 2022
For more information on Housing and Resident Life, click here.
For any questions related to QF Student Housing, please contact them at [email protected].
New Student Orientation (NSO) and Convocation
Your New Student Orientation (NSO) begins on July 17 and ends on July 21 with a formal convocation ceremony. It is mandatory for all incoming students to attend their NSO as you will meet all the people that play a role in your academic journey at Georgetown. Why is this important? Well, university life is very different from high school and you will be expected to perform at a faster pace from your first day on campus. We have a lot of resources to help students manage the transition to university life and it will save you lots of time in the future if you get to know who they are, where they are located and how they can help you shine!
New Student Convocation will be on Thursday, July 21, 2022 and is mandatory for all new students. The dress code is business or traditional attire. Please be respectful of cultural norms. We encourage parents and family members to gather and celebrate the beginning of your Georgetown career together.
International Students
Process and Requirements to Get a Visa for Qatar
If you do not have a Qatari ID or residency permit, GU-Q will process a student entry visa for you and email it to you. You cannot enter Qatar on a visitor visa in order to study, and the only required document for GU-Q to begin the process is a valid passport. The visa will be emailed to you by GU-Q around late June.
Traveling to Qatar
- Once you have recieved your visa and entry permit, you can book your ticket. Please do not book your travel until we have notified you that your visas and entry permits have arrived.
- After you have booked your ticket, upload your flight itinerary via your Application Status Page in order to issue an OK to board on your flight.
- Print out your visa and take it with you to the airport (an electronic copy will not suffice). It may be necessary to present a copy at passport control.
Getting your Residency Permit (RP)
After you arrive in Qatar, we will begin the process of obtaining a residency permit (RP). The RP allows you to stay in Qatar for more than 30 days and facilitates other processes, such as getting a national health card, a driver’s license, etc. The process includes:
- Upload Documents: Upload a passport size photo and blood type document via your Application Status Page. Not having these items will delay the process.
- Passport collection: At New Student Orientation (NSO), GU-Q will collect your passport temporarily to submit it to Qatar’s immigration department to get your RP.
- Processing: The process of getting a residency permit takes approximately 4-6 weeks. You will not be able to leave Qatar until your RP process is complete, unless there is an emergency. Please do not plan to travel for at least one month after your arrival.
- Getting Medical Clearance: To study and reside in Qatar, immigration laws require that individuals seeking residency be screened and test negative for HIV, Hepatitis B, Hepatitis C, and Tuberculosis. The issuance of the residency permit is contingent on passing the medical clearance process. We strongly recommend having these tests done in your home country prior to your arrival to Qatar. Positive findings may result in immediate repatriation from the country.
For more useful information for international students, click here.
For questions please contact The Office of Student Life.
It is easy to buy a SIM card with a Qatar number in Doha. You can add time by buying ‘top-up’ cards or buy a monthly plan. The two main vendors are Ooredoo and Vodafone. Please review their Qatar websites before your arrival so you can get connected quickly. Both Ooredoo and Vodafone have dedicated kiosks in the Arrivals Hall at Hamad International Airport and in major shopping malls.
GU-Q will assist international students with transportation from the airport if you provide your arrival information. For this reason, you must keep all contact information up-to-date and inform us of any changes to your flight.
► All international students must fill out the Transportation Request Form and upload your flight itinerary to your Application Status Page.
Note: If you request assistance, instructions will be emailed to you at a later time.
Please note the following:
- GU-Q assists with transportation for students only. If you are traveling with parents/siblings/companions, they will have to arrange their own transportation.
- We will only be providing transportation from the airport to QF Student Housing. If you plan to stay anywhere else besides QF Student Housing, you will have to arrange your own transportation.
Deadline to request transportation is July 1, 2022.
For questions, please contact the Office of Student Life.
GU-Q hosts mandatory orientation sessions for international students during the orientation week from July 17- July 20. During the orientation we will host interactive information sessions and social activities on academic life, immigration regulations, health care and cultural awareness of living in Qatar.
We require you to be in Qatar to attend this orientation.
Academic Related Information
Bachelor of Science in Foreign Service (BSFS) Degree Requirements
Your advising dean will explain the degree requirements when you arrive.
For more information on the BSFS degree requirements, click here
The Core Curriculum (“the Core”) offers students a deep foundation on which their major and elective courses are built while instilling values of citizenship and service. GU-Q students take the SFS Core (Proseminar, Government, History, Economics, Science and Map of the Modern World) in addition to the University Core requirements (Theology, Philosophy, Writing, HALC and Engaging Diversity). Together these courses give students the knowledge they need to understand and solve problems while maintaining the broad nature of a liberal arts education. For more information about the Core-Curriculum – First Two Years, click here
New students at GU-Q must submit fall course requests over the summer by filling in this form.
For more information on how to pre-register for courses, click here
In order to earn the BSFS degree, every student in the School of Foreign Service must demonstrate that he or she has the minimum skills necessary to complete academic or professional work in a modern language other than English. For more information about the Foreign Language Proficiency, please click here
To view the current Academic Calendar, please click here
To view the Undergraduate Bulletin, please click here
Academic Resources
To learn more about academic accommodations, please click here
To learn more about academic support services, please click here
Useful Information
To learn more about IT services, please click here
To learn more about facilities, please click here
To learn more about finances, please click here
To learn more about student employment, please click here
Want to know more? Click here for more about GU-Q academics, activities, and general student life. | https://www.qatar.georgetown.edu/admissions/new-students-information/ |
The image above is an actual image captured in our clinic by our equipment and rendered in office.
The orange "stuff" in the image above is the retina which is the photoactive tissue lining the inside of our eyeballs. It is extremely important to eyesight, and is very fragile. Because humans only have one small hole allowing light into the eye (the pupil) it is very difficult to visualize the retina in most circumstances. The most basic retinal screening would be when you utilize flash photography and the subject of your photo has "red eye". The red is actually a reflection of the retina bouncing out of the pupil, and although you can't tell much from this, at least you can say that your subject has a retina. To see the retina in any real detail can be challenging as the pupil is naturally small and gets even smaller when light is shone into it. This is why when there is any concern with the retina, an eye must be dilated (put drops in to make the pupil larger), allowing a better view of the tissue inside. A dilated retinal exam is still the gold standard for viewing the retina, but the problem is that it's time consuming, can cause certain types of glaucoma in at-risk people, and patients don't particularly enjoy being light sensitive and unable to focus up close for 4-6 hours while the drops are wearing off. This is why dilation is not routinely done on screening eye exams. Therefore it's important to have different ways to views the retina when the eye is undilated to help screen for cases where dilation is warranted.
The patient's eyeball in Figure 1 (above) has been routinely imaged in our clinic since 2004, and as you will see below, the way we examined and recorded the results (when undilated) has change drastically in the last 15 years, a time frame which we can divide it four major eras of imaging technology. At the start of each era, our clinic was consistently one of the country's very earliest adopters of each new tech.
analog era
pre - 2005
digital era
2005 - 2011
3D era
2011 - present
ultrawide era
2018 - present
THe Analog era (Pre - 2005)
A handheld light called an ophthalmoscope is shone into the eye. The device creates a small spotlight on the retina, and the doctor needs to move the spot around to view a larger swath of tissue. Due to the device lacking any imaging capabilities, any pertinent findings are usually recorded via a crude sketch. When your family doctor looks into your eye, this is what they are doing. Due to being cost effective and portable, the hand held opthalmoscope is still very popular in clinics worldwide. The drawing on the right represents the results of undilated opthalmoscopy on our patient from Figure 1, and as you can see, the results of which show little resemblance to each other (though not technically inaccurate).
Area of retina seen: 20° (after numerous sweeps of the light)
The Digital era (2005 - 2011)
The acquisition of a colour retinal camera was a significant step forward for us, not only in diagnostic capabilities but also in accuracy of record keeping. Having one colour image of some very important parts of the retina to be able to exam as close as needed, and to refer back to at a later date, was at the time truly amazing.
The retinal tissue captured in this image is starting to resemble the retina in Figure 1, and a trained clinician would be able to see similar blood vessels and structures between the two. However the field of view captured in this image is still fairly small as edge-to-edge in this image only represents about a 30° area of the retina. All the black surrounding the image is more retina that is unable to be seen by this technology. For reference, the yellow circle in the centre of this image is the optic nerve, and when you cross-reference the optic nerve to Figure 1, you can see how zoomed in this photo is.
Area of retina seen: 30° in a colour image.
The 3D Era (2011 - present)
Currently, we are still in the Topcon 3D OCT-2000 era and while this technology doesn't expand our field of view any further past the 30° mark, it does have the amazing ability to gather sub-surface, depth, and elevation data in this zone. In addition, it is able to measure and track the size of certain structures automatically, while cross referencing these value to a normative database to flag any concerns.
Area of retina seen: 30° in a colour image that includes sub-surface, depth, and elevation data.
The ultra-wide field era (2018 - present)
With the 3D era being so revolutionary in terms of diagnosing and tracking any changes in a specific area of the retina, the ultra-wide field era has been created to supplement this data by allowing us to venture, for the first time, to parts of the retina located outside of the historical 30° zone. In the image above, the same area of the retina is represented before and after the installation of the Zeiss Clarus 500 ultra-wide field camera. It's quite amazing how much more of the retina this camera can capture than it's predecessors, all while maintaining true colour, which is important in being able to diagnose concerning freckle-like lesions. In addition this camera also has the ability to show areas of imminent cell death by using a feature called 'fundus auto-fluorescence' which is remarkable.
The image used in Figure 1 is this ultra-wide montage photo collected by the Zeiss Clarus 500, combined with data from the Topcon 3D OCT, and rendered in-office. | https://calgaryvisioncentre.com/evolution |
A medium-range numerical weather prediction model operated by the United Kingdom METeorological Agency.
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Ultraviolet radiation
Electromagnetic radiation with wavelengths shorter than visible light, but longer than x-rays. Exposure to too much UV radiation can cause skin cancer.
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Unsettled
Changeable or variable weather.
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Unstable air
Air in which static instability exists. This condition is determined by the vertical gradients of air temperature and humidity.
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Updraft
A small-scale current of rising air. If the air is sufficiently moist, then the moisture condenses to become a cumulus cloud or an individual tower of a towering cumulus or Cb.
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Updraft base
Alternate term for a rain free base.
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Upper level
The portion of the atmosphere above the lower troposphere. It is generally applied to levels above 850 hPa.
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Upper level system
A general term for any large-scale or mesoscale disturbance capable of producing upward motion (lift) in the middle or upper parts of the atmosphere. This term is sometimes used interchangeably with impulse or shortwave.
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Upslope flow
Air that flows toward higher terrain, and hence is forced to rise. The added lift often results in widespread low cloudiness and stratiform precipitation if the air is stable, or an increased chance of thunderstorm development if the air is unstable.
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Upslope fog
Fog that forms when warm moist air is forced up a slope by wind.
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Upstream
Toward the source of the flow, or located in the area from which the flow is coming.
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Upwelling
The rise of colder, deep ocean waters along a coast due to the movement of surface water away from the coast.
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UTC
Abbreviation for Universal Time Coordinate. See also Zulu, GMT.
Weather News
Flood-ravaged NSW holiday park residents struggle to rebuild a month later
21:07 EST
A month after severe flooding, residents of a northern New South Wales holiday park are struggling to rebuild and some desperately need help.
Far North QLD soaked by Melbourne's annual rainfall in three days
15:56 EST
Parts of Far North Queensland have received more than 800mm of rain during the last four days, with some places receiving Melbourne's entire annual average rainfall in just 72 hours. | https://www.weatherzone.com.au/help/glossary.jsp?l=u |
Milwaukee Chamber Orchestra will present a free community tribute concert on Saturday, January 14, 2012 at 3:00 pm at the Holy Redeemer Institutional Church of God in Christ, in Milwaukee. Music Director Richard Hynson will conduct, and church founder Bishop Sedgwick Daniels will narrate the performance.
The Dream Lives On will feature works for string orchestra including Dmitri Shostakovich’s heart-wrenching Chamber Symphony, as well as selections from Robert Ray’s Gospel Mass performed by singers from the Bel Canto Chorus of Milwaukee. The tribute seeks to underscore the solemnity of the occasion and Dr. Martin Luther King, Jr.’s non-violent transcendence of the brutal divisiveness of his times.
Reflecting on the significance of this event, Hynson said, “Dr. King’s message of justice, freedom, and equality speaks to us today as vividly as it did years ago. His clarion call for unity and mutual acceptance urges us to reach across the boundaries of fear and mistrust to join together in building an America free from prejudice. What better way to symbolize that joining together than for us to come to this house of prayer to honor Dr. King and his legacy through music and words?”
Concertgoers are encouraged to reserve free seats in advance by visiting www.milwaukeechamberorchestra.org and clicking on “Reserve Your Tickets Now.” Seating will also be available on the day of the event. Doors will open at 2:15 pm.
The Milwaukee Chamber Orchestra is Milwaukee’s only professional chamber orchestra and one of only 65 in the nation. Since its founding in 1973, the MCO has earned a reputation as one of southeastern Wisconsin’s finest professional performing arts groups. Critical review, audience response and other feedback reveal that the MCO performs with the highest of artistic integrity and fills a unique niche in satisfying a desire for audiences to hear, and for professional artists to perform, chamber orchestra music within the greater Milwaukee metropolitan area.
Bel Canto Chorus is Milwaukee’s premier independent choral organization. Founded in 1931, Bel Canto is committed to enriching the lives of its audiences and singing members through the outstanding live presentation of the finest choral music. The chorus performs four unique concerts each season in Greater Milwaukee’s most beautiful venues.
Richard Hynson has served as music director of the Milwaukee Chamber Orchestra since 2006. Additionally, he has served as music director for Bel Canto Chorus since 1987. In demand as a guest conductor, Hynson’s past engagements include performances with the Milwaukee Symphony, the Skylight Opera Theatre, and the Racine, Sheboygan, and Waukesha Symphony Orchestras. Hynson has conducted at Carnegie Hall in New York City, where he led a large national festival chorus and orchestra in Ralph Vaughan Williams’s Dona nobis pacem. In the summer of 2008, Hynson conducted the string orchestra and chorus for the Prelude Music Academy summer camp in Madison. In 2009, he guest-conducted the Wisconsin Chamber Orchestra in one of the Concerts on the Square in Madison. He also served as the Music Director for Gathering on the Green, the popular outdoor music festival in Mequon, WI, in 2009 and 2010. In addition to his work as a conductor and educator, Hynson is a composer. He has written a substantial body of published choral, vocal, and ensemble works. Music critic Elaine Schmidt praised Hynson’s most recent performance with the Milwaukee Chamber Orchestra, in their collaborative production of Astor Piazzolla’s Maria de Buenos Aires: “Hynson and the ensemble supported the singers gracefully and created a get-under-your-skin sonic backdrop that did more to set the story in Buenos Aires than detailed scenery could have managed” (Schmidt, Milwaukee Journal Sentinel, November 11, 2011). | http://milwaukeecourieronline.com/index.php/2012/01/14/milwaukee-chamber-orchestra-presents-the-dream-lives-on-a-tribute-to-dr-martin-luther-king-jr/ |
Easy Chickpea and Cauliflower bowl with Moroccan spices and a mint cilantro herb sauce. This bowl is delicious, springy, zesty and comes together within minutes.
Prep the cauliflower and put it in the oven. Spice up the chickpeas. Get the greens and veggies ready. Make the mint cilantro herb sauce. Layer up the bowl as you wish. Drizzle herb sauce. Add some non dairy yogurt to balance out the spices if you wish. and serve.
Use all mint or all cilantro if you have a problem with either of the 2 herbs. Add any roasted veggies to the bowl for variation. The time is mainly to collect the ingredients and spices, chop them up and get going. Active time after that is 15 minutes. You can make the herb sauce and keep it ready to throw over roasted veggies or chickpeas.
This bowl will definitely help finish up all those chickpeas you have been saving after the chickpea brine train 😉
More Chickpea recipes from the blog
- Sweet Potato Chickpea Peanut Burgers.
- Teriyaki chickpea Bao – Steamed buns
- Chickpea Stuffed Poblano Peppers with Smoky Tomato Sauce.
- Buffalo Chickpea Pizza with celery Ranch
- Potato Chickpea Cauliflower Salad with Sour cream onion dressing
Bake the cauliflower, spice up the chickpeas. Layer up with greens, tomatoes and other juicy veggies.
Drizzle generously with herb sauce and some non dairy yogurt (optional)
Moroccan Chickpea Cauliflower Bowl with Herb sauce.
Ingredients
For the cauliflower:
- 1 tsp cumin seeds
- 1 head of cauliflower
- 2 tsp oil
- 1/2 tsp (0.5 tsp) salt
- 1/2 tsp (0.5 tsp) cayenne
- 1/4 to 1/2 tsp garlic granules
- 2 tablespoons breadcrumbs optional
For the chickpeas:
- 15 oz (425.24 g) can chickpeas or 1.5 cups cooked
- 1 tsp oil
- 1/2 (0.5 ) or more salt depending on if the chickpeas were salted
- 3/4 tsp (0.75 tsp) cumin powder
- 1/4 to 1/2 tsp cayenne
- 1/2 tsp (0.5 tsp) paprika
- 1/4 tsp (0.25 tsp) garlic granules
- 1/4 tsp (0.25 tsp) cinnamon
- 1/2 tsp (0.5 tsp) lime juice
Mint Cilantro Herb sauce:
- 1/2 cup (22.5 g) mint
- 1/4 cup (11.25 g) cilantro or use more mint
- 1 tsp olive oil
- 1 to 2 tsp lime juice
- 2 to 4 tablespoons water
- 1/4 tsp (0.25 tsp) cumin powder
- 1 clove of garlic
- 1/4 tsp (0.25 tsp) salt
- 1/4 tsp (0.25 tsp) sugar or maple
For the bowl:
- Spinach greens, tomatoes, 2 tbsp non dairy yogurt (optional) etc
- slivered almonds and pepper flakes for garnish
Instructions
- Toast the cumin seeds on a skillet over medium heat until fragrant. about 2 minutes. Grind into a coarse powder. Toss the cauliflower florets in oil or spray oil to coat all the florets. Rub a little with your hands to coat. Add salt, cumin and the spices and mix well to coat. Add breadcrumbs if using and toss. Place on baking sheet and bake at 425 degrees F / 220ºc for 25 to 30 minutes.
- Add chickpeas and all the ingredients except lime juice in a skillet. Cook over medium heat for 3 to 4 minutes. Add lime juice and toss.
- Make the herb sauce: Blend everything until well combined. taste and adjust salt and sweet. (Use all mint or all cilantro if you have an issue with one of them).
- Arrange the roasted cauliflower, spiced chickpeas, greens and veggies. Drizzle the herb sauce. Drizzle non dairy yogurt if using. Add red pepper flakes, slivered almonds. Serve. | https://www.veganricha.com/2015/03/moroccan-chickpea-cauliflower-bowl-with-herb-sauce.html |
Years of steadily accumulating dust and grime had taken a toll on King Tut’s tomb, but a recently completed restoration project has revitalised the historic chamber, while making much-needed infrastructure improvements to prevent ongoing decay.
The 3,350-year-old tomb is located in the Valley of Kings near the Egyptian city of Luxor. Rock-cut steps lead down some 40 feet to four chambers, of which only Tut’s burial chamber is adorned with painted walls. Most of the chamber’s relics were removed after its discovery in 1922, but some artefacts remain, including the quartzite sarcophagus and its granite lid, the gilded wooden coffin exterior, and the man himself, whose mummy is on display in an oxygen-free case.
Intense interest in the tomb has caused problems, however. Dust delivered to the tomb by tourists cast a grey veil over the walls. Concerns emerged that increased levels of carbon dioxide and humidity were stimulating microbial growth, including some worrisome brown splotches on the paintings. Some areas of the chamber exhibited scratch and scuff marks caused by tourists and film crews. The microclimate inside the chamber was also unpleasant for the visiting tourists. The tomb was cramped and crowded, poorly lit, and devoid of helpful signage. | https://www.gizmodo.co.uk/2019/01/see-inside-the-newly-renovated-king-tuts-tomb |
Valley of the Queens, also known by the names Biban el-Sultanat, Biban el- Harim and Wadi el-Malikat is located on the west bank of the river Nile in Egypt. The necropolis has more than seventy tombs, most of which are very lavishly and beautifully decorated. The tomb where Queen Nefertari, who lived from 1290-1224 BCE, is a true example of distinct beauty built in the 19th dynasty.
Who was Nefertari?
Nefertari Merytmut, which means Beautiful Companion or Beloved of Mut, lived in the 19th dynasty and was the royal wife of Ramesses I. She is the best known of Egyptian queens, just after Cleopatra. The queen was adorned and honored with many titles during her life some of which were Sweet of Love, Great of Praises, Great King’s wife, Lady of the Two Lands and many others. Nefertari and Ramesses are said to have at least six children, four sons and two daughters.
Tomb of Nefertari
Referred as QV66, Nefertari’s tomb is among the most spectacular, lavishly and beautifully decorated tombs located in the Valley of the Queens. The tomb was first discovered in 1904 by Ernesto Schiaparelli. The tomb has a long staircase beginning from the entrance and ending into a hall that is 17x17.5 feet. The hall contains rock cut bench and cavetto cornice to keep funerary items.
The tomb QV 66 resembles a house and has intimacy which is rare to be found in any ancient tomb of Egypt. There are pictures from the queen’s life and her journey hereafter. Also, the walls of the burial chamber have poetry which was written by Ramesses II for his beloved wife. Since Nefertari was not a Pharaoh, her tomb QV 66 does not contain any images from her daily life. Also, the text found on the walls was very restricted. The text that was found on the walls was from the Book of the Dead.
Temple of Nefertari in Abu Simbel
The Temple of Nefertari is the smaller of the two temples that were built by Ramesses II in Abu Simbel. Built in the honor of Nefertari and Goddess Hathor, the temple is the second of its kind in Egyptian history which was dedicated to a queen. The temple has a number of statues, images, inscriptions and hieroglyphics showing Ramesses II’s heroic acts and Nefertari paying homage to the gods are present.
Present day situation
The tomb was unfortunately robbed for all its treasures by tomb robbers. The mummy and sarcophagus too of Nefertari were stolen. Some pieces of the mummy which were found at the tomb are on display at Turin in the Egyptian Museum.
The tomb of Nefertari was opened for public in the year 1995, but was again closed in 2003 due to the preservation of the place. People on specialized tours however are still allowed with special permits. Even the limited numbers of tourists have a consequence on the surface of the paintings. Their moist bacteria-laden breath causes mould to grow on the surface; the tomb is after all a closed environment. Thus, to preserve the same no visitors are allowed in. | https://www.ask-aladdin.com/Egypt-Sites/Egypt-tombs/nefertari.html |
Practical review on the use of synchrotron based micro- and nano- X-ray fluorescence mapping and X-ray absorption spectroscopy to investigate the interactions between plants and engineered nanomaterials.
The increased use of engineered nanomaterials (ENMs) in commercial products and the continuous development of novel applications, is leading to increased intentional and unintentional release of ENMs into the environment with potential negative impacts. Particularly, the partition of nanoparticles (NPs) to waste water treatment plant (WWTP) sludge represents a potential threat to agricultural ecosystems where these biosolids are being applied as fertilizers. Moreover, several applications of ENMs in agriculture and soil remediation are suggested. Therefore, detailed risk assessment should be done to evaluate possible secondary negative impacts. The impact of ENMS on plants as central component of ecosystems and worldwide food supply is of primary relevance. Understanding the fate and physical and chemical modifications of NPs in plants and their possible transfer into food chains requires specialized analytical techniques. Due to the importance of both chemical and physical factors to consider for a better understanding of ENMs behavior in complex matrices, these materials can be considered a new type of analyte. An ideal technique should require minimal sample preparation, be non-destructive, and offer the best balance between sensitivity, chemical specificity, and spatial resolution. Synchrotron radiation (SR) techniques are particularly adapted to investigate localization and speciation of ENMs in plants. SR X-ray fluorescence mapping (SR-XFM) offers multi-elemental detection with lateral resolution down to the tens of nm, in combination with spatially resolved X-ray absorption spectroscopy (XAS) speciation. This review will focus on important methodological aspects regarding sample preparation, data acquisition and data analysis of SR-XFM/XAS to investigate interactions between plants and ENMs.
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Excited about Apollo and want to make it better? We’re excited too. Read below to learn about different ways to contribute to the project.
A great place to start if you're looking for ideas on how to contribute, guidance through our contribution workflow, or discussion with other contributors, is the #contributing channel on Apollo Slack. We're always looking for new library maintainers and issue triagers, and this channel is the best place to see where your help is needed.
Every week, Apollo's core contributors and maintainers meet to review project issues and pull requests and discuss work on the latest project milestones. You can review minutes from every meeting here and request an invitation to discuss your PR or project idea in the #contributing channel.
If you encounter a bug, please file an issue on GitHub via the repository of the sub-project you think contains the bug. If an issue you have is already reported, please add additional information or add a 👍 reaction to indicate your agreement.
Intended outcome: What you were trying to accomplish when the bug occurred, and as much code as possible related to the source of the problem.
Actual outcome: A description of what actually happened, including a screenshot or copy-paste of any related error messages, logs, or other output that might be related. Places to look for information include your browser console, server console, and network logs. Please avoid non-specific phrases like “didn’t work” or “broke”.
Reproduction: A minimal reproduction of the issue that we can run. This is ideally in the form of a small application we can clone from GitHub, or a failing test sent in the form of a PR. The reproduction should contain as little code as possible to demonstrate the bug.
Creating a good reproduction really helps contributors investigate and resolve your issue quickly. In many cases, the act of creating a minimal reproduction illuminates that the source of the bug was somewhere outside the library in question, saving time and effort for everyone.
Use case: What are you trying to accomplish, in specific terms? Often, there might already be a good way to do what you need and a new feature is unnecessary, but it’s hard to know without information about the specific use case.
Could this be a plugin? In many cases, a feature might be too niche to be included in the core of a library, and is better implemented as a companion package. If there isn’t a way to extend the library to do what you want, could we add additional plugin APIs? It’s important to make the case for why a feature should be part of the core functionality of the library.
Is there a workaround? Is this a more convenient way to do something that is already possible, or is there some blocker that makes a workaround unfeasible?
Feature requests will be labeled as such, and we encourage using GitHub issues as a place to discuss new features and possible implementation designs. Please refrain from submitting a pull request to implement a proposed feature until there is consensus that it should be included. This way, you can avoid putting in work that can’t be merged in.
Once there is a consensus on the need for a new feature, proceed as listed below under “Big PRs”.
In addition to reporting issues, a great way to contribute to Apollo is to respond to other peoples' issues and try to identify the problem or help them work around it. If you’re interested in taking a more active role in this process, please let us know on the Apollo Slack!
If there is a small change to be made, please feel free to open a PR right away with the fix or improvement. For this category of pull requests, there is no need to write a design first.
Improving the documentation, examples, and other open source content can be the easiest way to contribute to the library. If you see a piece of content that can be better, open a PR with an improvement, no matter how small! If you would like to suggest a big change or major rewrite, we’d love to hear your ideas but please open an issue for discussion before writing the PR.
For a small bug fix change (less than 20 lines of code changed), feel free to open a pull request. We’ll try to merge it as fast as possible and ideally publish a new release on the same day. The only requirement is, make sure you also add a test that verifies the bug you are trying to fix.
For significant changes to a repository, it’s important to settle on a design before starting on the implementation. This way, we can make sure that major improvements get the care and attention they deserve. Since big changes can be risky and might not always get merged, it’s good to reduce the amount of possible wasted effort by agreeing on an implementation design/plan first.
Open an issue. Open an issue about your bug or feature, as described above.
Reach consensus. Some contributors and community members should reach an agreement that this feature or bug is important, and that someone should work on implementing or fixing it.
Agree on intended behavior. On the issue, reach an agreement about the desired behavior. In the case of a bug fix, it should be clear what it means for the bug to be fixed, and in the case of a feature, it should be clear what it will be like for developers to use the new feature.
Agree on implementation plan. Write a plan for how this feature or bug fix should be implemented. What modules need to be added or rewritten? Should this be one pull request or multiple incremental improvements? Who is going to do each part?
Submit PR. In the case where multiple dependent patches need to be made to implement the change, only submit one at a time. Otherwise, the others might get stale while the first is reviewed and merged. Make sure to avoid “while we’re here” type changes - if something isn’t relevant to the improvement at hand, it should be in a separate PR; this especially includes code style changes of unrelated code.
Review. At least one core contributor should sign off on the change before it’s merged. Look at the “code review” section below to learn about factors are important in the code review. If you want to expedite the code being merged, try to review your own code first!
Required CI checks pass. If the tests don’t pass, there is no need to review the PR until they do.
Simplicity. Is this the simplest way to achieve the intended goal? If there are too many files, redundant functions, or complex lines of code, suggest a simpler way to do the same thing. In particular, avoid implementing an overly general solution when a simple, small, and pragmatic fix will do.
Testing. Do the tests ensure this code won’t break when other stuff changes around it? When it does break, will the tests added help us identify which part of the library has the problem? Did we cover an appropriate set of edge cases? Look at the test coverage report if there is one. Are all significant code paths in the new code exercised at least once?
No unnecessary changes. PRs shouldn’t come with random formatting changes, especially in unrelated parts of the code. If there is some refactoring that needs to be done, it should be in a separate PR from a bug fix or feature, if possible.
Code has appropriate comments. Complicated logic should be commented, or written in a clear “self-documenting” way.
Idiomatic use of the language. In TypeScript, make sure the typings are specific and correct. In ES2015, make sure to use imports rather than require and const instead of var, etc. Ideally a linter enforces a lot of this, but use your common sense and follow the style of the surrounding code. | https://www.apollographql.com/docs/community/contributing/ |
Meet potential employers and find out about jobs and internships at graduate recruitment events.
Graduate recruitment events include a range of activities:
- Careers Expos
- Employer Presentations and Information Sessions
- Industry and Networking Events
- On-campus interviews and assessments
Benefits of participating in graduate recruitment
There are many good career development reasons for taking part in graduate recruitment activities, besides finding internships or employment after graduation. These include being able to:
- meet with recruiters in an informal yet professional environment
- learn about the type of work being offered across a range of options
- hear about the experiences of recent recruits
- find out how organisations differ
- discover the skills and attributes valued by employers
- gain exposure to potential employers and establish a personal relationship before making an application
- get familiar with the language of the workplace and graduate recruitment
- develop the confidence required to apply for a range of professional opportunities
How to get involved
Graduate recruitment activities happen from the beginning of and throughout the academic year. Even if you are not in your final year of study, you can: | https://www.wgtn.ac.nz/careers/employment/graduate-recruitment |
Rock, Love Ballads, Blues, hint of Jazz, an eclectic combination with traditional native tunes.
11 MP3 Songs
EASY LISTENING: Adult contemporary, POP: 60''s Pop
Details:
Performing regionally in the Southwest has developed Willow Pony for the release of their long awaited album, Children of the Sun. Jicarilla Apache songwriter, Fred Vigil has become a producer as well. The title song won the 2006 Native American Category, and was performed at the awards banquet, a first in the 19 year history, which included: Adrian Wall-native flute, Mike Swick-Bass, Susan Clark-Keyboards/vocals, and Fred Vigil-guitar/vocals.
The songs provide a glimpse of modern day living in the Southwest, and offers unique perspectives which are global. Black Eagle-Grammy Award Winning group closes the album.
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Changes are on the horizon for the way Americans can safely and securely return their medication for environmentally responsible disposal. The U.S. Drug Enforcement Administration (DEA)'s final rule on the collection of controlled substances, such as Oxycontin, Percocet, and Vicodin-released September 9, 2014-will eventually give consumers much greater flexibility as to when, where, and how they can dispose of their unwanted medication. The rule allows manufacturers, distributors, reverse distributors, narcotic treatment programs, retail pharmacies, and hospitals/clinics with a retail pharmacy to voluntarily apply for authorization to and maintain on-site drug collection receptacles and provide mail-back programs. Law enforcement agencies may also continue to collect controlled substances, and may also now offer mail-back programs. While the rules "go into effect" October 9, 2014, consumers won't be able to start dropping off their medicines right away. State and municipal governments, local law enforcement agencies, state boards of pharmacy, pharmacy retailers, manufacturers, reverse distributors, and a host of other stakeholders will need to take a number of steps to comply with and carry out the rules. This includes everything from modifying individual state pharmacy and drug disposal regulations to securing additional funding to support the expanded drug take-back programs. Featuring a diverse and dynamic panel of thought leaders and pharmaceutical stewardship experts, this webinar will explore the health, environmental, technical, and financial implications of the new DEA rules for consumers, governments, law enforcement, pharmacies, manufacturers, and reverse distributors. In particular, it will examine the unique challenges and potential solutions to financing the expanded drug take-back opportunities that the new rule allows. There will be ample time for Q+A.
Speaker bios:
Pat Perry is the Director of Environmental Management at CVS Health. She leads the Environmental Department, responsible for compliance across the enterprise. Ms. Perry is well-versed in regulatory requirements applicable to retail under the CWA, CAA, EPCRA, and in particular, requirements for management of retail and pharmaceutical waste under RCRA. Prior to CVS Health, she led the Environmental Department of another major retailer for 15 years. Her experience and talent provide a unique perspective on how to drive successful programs in a retail setting.
Ed Gottlieb, Industrial Pretreatment Coordinator at the Ithaca Area Wastewater Treatment Facility, Chairs the Coalition for Safe Medication Disposal in Tompkins County, N.Y. He organized numerous one day collection events and inventories of returned medications. He created a series of how-to videos to help others organize collection events. Ed led the successful effort to establish a county-wide drop box program. He has been an active member of the national pharmwaste listserv, contributing extensively on the topic of the DEA rule making.
Charlotte Smith is a consultant in the area of pharmaceutical waste management and Senior Regulatory Advisor, PharmEcology Services, a business unit of Waste Management Healthcare Solutions, Inc. (WMHS), a Waste Management company. She founded PharmEcology Associates, LLC in 2000 and sold the company to WMHS in 2009. She co-founded Capital Returns, Inc., a nationally known pharmaceutical reverse distributor in 1991 and for 10 years served as president and chief regulatory advisor. Ms. Smith is also a registered pharmacist.
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Object of the Week: Spear Thrower
In his bronze sculpture of a Spear Thrower, Paul Manship depicts an athlete in motion. The sculpture has balance and equilibrium as the figure reaches back and prepares to hurl his spear forward. He’s pictured at the moment just before the energy is transferred, with his full weight on the back leg, where the muscles bunch and bulge with exertion. If he’s hurting from the effort, his face doesn’t show it; his look is one of resolve and otherworldly gracefulness. His spear creates a strong horizontal line that is carried across the sculpture by the figure’s fully extended left arm. His features are ripped and generalized; he is an ideal form and not an individual one. With this sculpture Manship celebrated ideas like human strength and achievement, and the beauty of the athlete’s body.
Important links between works of visual art exist everywhere, and part of what SAM and other art museums can do for us is point out this vast web of interconnectedness. Its intricacy and complexity mean that there is always more to discover. For instance, Paul Manship, an American born in Minnesota, was inspired by Indian art, as well as archaic Greek art and the Italian Renaissance that renewed appreciation for classical Greek ideas and developed them further.
In the form of Spear Thrower Manship made a direct reference to this Bronze statue of Zeus or Poseidon at the National Archaeological Museum in Athens. Done in what’s called the “severe style” around 460 BCE, it is a true landmark in the history of art, and remains one of the best-known examples of Early Classical Greek sculpture. It was to this school of art-making, and to this particular work, that Manship looked when he cast the Spear Thrower in 1921.
Interesting comparisons, if less apparent ones, exist even in SAM’s own collection. Have a look at our Black-Figure Amphora displayed on the fourth floor, amid other works from the ancient Mediterranean. Revelers stride across the scene in dynamic poses that have them twisting and contorting their bodies in displays of balance and gracefulness. Each figure stands on a single foot, supported by a powerful, muscular leg. Clean, sinuous lines mark the contours of the figures. All these traits surface visibly in Manship’s work of 2,300 years later.
We’d be doing Manship a disservice, though, if we understood him as only looking backwards. In SAM’s Spear Thrower, as in Manship’s famous Prometheus fountain at Rockefeller Center, he innovated a combination of classical, idealized bodies and a distinctly Modern, streamlined aesthetic that secured him a prominent place in the web of art history. | https://samblog.seattleartmuseum.org/2017/02/spear-thrower/ |
Treat the numbers or treat the patient?
- Victor M Montori
- Aust Prescr 2011;34:94-5
- 1 August 2011
- DOI: 10.18773/austprescr.2011.054
Current practice guidelines, regardless of healthcare system and country of origin, increasingly carry a similar message: treat to target. These targets are often expressed in terms of laboratory parameters which are presumed to reflect the control of a patient's condition, and by extension their health and prognosis. The assumption is that 'normalising' parameters, such as lipids, blood pressure and blood glucose in patients with type 2 diabetes, will lead to better outcomes. However, these parameters are surrogate outcomes and do not guarantee long-term clinical benefits.
Is the assumption of benefit from intense control of these parameters based on high quality evidence? Some evidence suggests that there is no benefit, but there may be marginal harm associated with intensive control of risk factors in patients with diabetes.1-3 Benefits may still accrue for younger less sick patients, but this remains speculative. Even if true, those benefits would have to offset the downside of treatment, a task made difficult by the relative good health of the patients and the necessarily prolonged course of treatment. In these younger and healthier patients, intense lifestyle modification – smoking cessation, diet, exercise, stress reduction – may be more compelling than intensive drug treatment.
Treat-to-target often requires clinicians to prescribe more drugs at higher doses. This in turn calls for more laboratory testing to determine the efficacy of these interventions on the parameters of interest and to monitor the safety of the drugs on the patient's body. Treat-to-target requires patient self-monitoring and self-management in response to the monitoring results plus more visits to nurses and physicians. Higher doses and combination therapy may also increase the likelihood of adverse effects, which in turn may require increased medical attention and reduce the patient's capacity to do patient work.
The increasing demand for treatments, investigations and visits will test the capacity of the patient and their caregivers to implement these complex programs. By some estimates, the work of being a patient with diabetes requires more than two hours every day.4 Patients are expected to prioritise this 'part-time job' – to understand, plan and enrol others to help with the plan, to implement and adhere to the plan, and to reflect and value the treatment enough to keep going day after day. They have to fit this around the work of being a parent, sibling, child, spouse or relative, the work of being an employee or boss, and the work of being a citizen, a hobbyist, or a sports player.
The extent to which the patient's other 'jobs' are flexible enough to accommodate the ever-increasing work of being a patient and the ability of patients to enrol others to assist with the tasks of 'patienthood' may vary over time. Eventually, the expansion of patient workload may exceed the capacity of the patient or their caregivers to accommodate its demands. This forces the patient to prioritise, compromise and do only part of the expected patient work. They may then appear to be non-adherent to treatment.
The clinician may notice this non-adherence as missed appointments, incomplete self-monitoring data and in test results that reflect poor control. The clinician at this point, under a treat-to-target approach, may feel obliged to intensify the therapy. This carries the unintended consequence of increasing the treatment workload, further overflowing the patient's capacity to execute the plan, with ongoing deterioration not only of disease control but also of the patient–clinician relationship.
Our research group is exploring how best to respond to this form of non-adherence, which reflects the constraints of many competing demands that patients face. What can clinicians do in the meantime? | https://www.nps.org.au/australian-prescriber/articles/treat-the-numbers-or-treat-the-patient |
Composting is a highly sustainable method of waste disposal. This process breaks down natural materials into smaller particles, which can then be used as soil and for other natural purposes.Adding compost to soil helps provide soil with extra nutrients. Compost has nitrogen, phosphorous, potassium, and micronutrients manganese, copper, iron, and zinc. This makes compost superior to commercial fertilizers which are often lacking micronutrients. Compost acts as a slow-release fertilizer since the organic materials all decompose at different times. Soil pH also benefits from compost because it neutral, helping to keep pH at the best level. | https://clockworkremovals.com.au/benefits-of-composting/ |
Georgia State University College of Law has claimed the No. 1 spot on a list of the nation’s best-value law schools.
The National Jurist explains why the college is at the top of its list. The average student debt of Georgia State University law students is $64,384 and its employment rate is 82.3 percent.
The top 10 law schools on the list are:
1) Georgia State University, $16,858 tuition
2) University of Georgia, $19,488 tuition
3) University of Florida, $22,299 tuition
4) University of Wisconsin, $21,450 tuition
5) University of Nebraska, $15,036 tuition
6) University of South Dakota, $14,501 tuition
7) University of Arkansas Fayetteville, $15,704 tuition
8) University of Alabama, $23,055 tuition
9) University of Kentucky, $22,700 tuition
10) University of Oklahoma, $19,973 tuition
The schools are ranked based on the percentage of students who pass the bar exam (weighted 15 percent), employment rate (weighted 35 percent), resident tuition (weighted 25 percent), cost of living (weighted 10 percent), and average indebtedness at graduation (weighted 15 percent). | http://elderlawlibrary.com/georgia-state-university-tops-list-of-best-value-schools/ |
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