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+{"text": "Extrapleural Solitary Fibrous tumors (SFTs) have been increasingly reported. The retroperitoneum, deep soft tissues of proximal extremities, abdominal cavity, trunk, head and neck are the most common extraserosal locations reported. Microscopically they show a wide range of morphological features, and so the differential diagnosis is extensive. Immunohistochemically, they commonly express CD34, vimentin, bcl-2 and CD99. Epithelial membrane antigen (EMA) and smooth muscle actin (SMA) may occasionally be expressed. Epithelioid morphology in extrapleural SFT has only very occasionally been described (five cases reported), some of them with biphasic pattern and others with malignant characteristics.A SFT of the thigh with epithelioid areas in a 63 year old woman is reported. Microscopically the tumor showed areas hypo and hipercellular. At the periphery of the hipercellular areas there were nodules composed of epithelioid cells. Immunohistochemically both the spindle and epithelioid cells were positive for CD34, vimentin, bcl-2 and CD99. Epithelial, neural and muscular markers were negative. Molecular study was done and ruled out a synovial sarcoma.Ten cases of SFT of the thigh have been reported but to our knowledge this is the first case with epithelioid morphology affecting the extremities. Identification of this pattern of SFT is of importance, to avoid misdiagnosis with other more aggressive conditions in soft tissue. Extrathoracic solitary fibrous tumors have been described at almost every anatomic location -3, but rA 63 year old woman presented with a three year history of a painless growing mass in the groin and increasing pain in her left leg during the last year. Magnetic resonance imaging (MRI) showed a lesion measuring 11 \u00d7 7 \u00d7 7 cm. near the left hip affecting the quadriceps. The lesion was suggestive of a soft tissue sarcoma. Total body scan revealed no distant metastasis. Excisional biopsy was done, resulting in a low-grade fusocellular tumor with hemangiopericytic pattern.Surgical treatment was carried out and the whole tumor was submitted for histopathological study. The patient was treated with radiotherapy (63 Gy). Control MRI, six months after radiotherapy, showed no lesions.Grossly the tumor appeared as an encapsulated, tan-grey mass measuring 10 \u00d7 4 \u00d7 3 cm. At cut section the tumor was pseudolobulated, with small hemorrhagic foci, and yellow necrosis. No differences in color and consistence were found between lobules.Microscopically, the tumor was composed of a proliferation of fusocelular cells with haphazardly distribution and varying degrees of stromal collagenization. The cellularity greatly varied in different areas with a predominance of hypercellular areas. Medium-sized thin-walled blood vessels in a hemangiopericytic growth pattern were observed, being more evident in hypercellular areas at the periphery of the lobules. Highly cellular spindle cell areas resembled fibrosarcoma and very occasionally multinucleated cells were seen. Mitosis were spare (\u2264 2 mitosis in 10 HPF) but foci of coagulative necrosis existed.At the periphery of the tumor and close to hipercellular areas we found three isolated and fairly well demarcated nodules each one measuring 12\u201315 mm, where cells adopted an epithelioid morphology with round, vesicular nuclei with micro nucleoli and abundant eosinophilic cytoplasm. These cells were mainly arranged in solid sheets but nests, pseudoglandular, or cleft patterns were also present. Focally artifactual shrinkage produced pseudovascular spaces. In these areas pleomorphism was moderate. No fusocellular cells were seen. Mitosis score was more than 10 in 10 HPF, some of them abnormal, and foci of necrosis were observed. A thin band of collagen isolated these epithelioid nodules from the fusocelular rich areas except in one of the nodules which both epithelioid and spindle areas merged in indistinct transition.Cells from fusocelular and epithelioid areas showed the same immunophenotype expression: vimentin +, CD34 +, CD99 +, and bcl2 +, being completely negative for epithelial, neural and muscular markers. Ki67 immunolabeling was low in fusocellular areas (< 5%) and rather high (> 40%) in epithelioid ones. (All antibodies from Dako)Polymerase chain reaction (PCR) for presence of a SYT-SSX1 or SYT-SSX2 fussion transcript  proved nExtrapleural solitary fibrous tumor, especially those at the extremities, still represent a rare entity of soft tissue neoplasms . In a cuHistologically SFT are well circumscribed, and consist of bland spindle cells showing a wide spectrum of histological features ranging from hypercellular to myxoid or hialinized pattern-less hypocellular areas. Hemangioperycitomatous pattern is also evident mainly in hipercellular areas of tumors. Mitoses are infrequent and necrosis is not common in SFT. Some histological variants have been described as giant cell SFT fibroma and fat forming SFT ,16. ImmuAlthough clusters of polygonal  cells have been described in SFT of pleura and mediastinum  the termMicroscopically, three of the cases reported showed a biphasic morphology coexisting areas of typical fusocellular SFT and areas of epithelioid appearance ,7, one oPathological criteria of malignancy include large tumor size (more than 50 mm), infiltrative margins, high cellularity, nuclear pleomorphism, areas of tissue necrosis and increased mitotic index (more than 4 mitosis in 10 HPF) ,3,19. InThe behavior of SFT is unpredictable . The relImmunohistochemical results on epithelioid SFTs are quite contradictory. These epithelioid cells usually retain an immunophenotype of SFT such as CD34 +, vimentin +, CD99 + and bcl-2 + -8, excepThe fusocellular component was positive for CKAE1-AE3 and MNF116 in one of the cases reported . The expIn our case, immunohistochemical stains demonstrated positivity of epithelioid cells with vimentin, CD34, bcl2 and CD99 and negativity for epithelial, neural and muscular markers.The differential diagnosis of SFTs of soft tissues is extensive. Hemangiopericytoma is one of the lesions that bears a close resemblance with SFT of soft tissues . The hemagioperycitomatous pattern is present in a wide variety of tumors; in fact, in absence of this pericytic differentiation , most heThe epithelioid growth pattern contributes to further difficulties in the differential diagnosis of these tumors with other tumors having epithelioid features, such as epithelioid sarcoma, schwannoma, leiomyosarcoma, MPNST, hemangiopericytoma and synovial sarcoma among others.The differential diagnosis with synovial sarcoma may prove very difficult, mainly if SFT express cytokeratin and EMA. Synovial sarcoma rarely expresses CD34. No molecular studies seeking a SYT-SSX1 or SYT-SSX2 fusion transcript have been performed in CK+ epithelioid SFT cases reported. In one case , the FIS"}
+{"text": "Short interfering RNAs (siRNAs) can be used to knockdown gene expression in functional genomics. For a target gene of interest, many siRNA molecules may be designed, whereas their efficiency of expression inhibition often varies.To facilitate gene functional studies, we have developed a new machine learning method to predict siRNA potency based on random forests and support vector machines. Since there were many potential sequence features, random forests were used to select the most relevant features affecting gene expression inhibition. Support vector machine classifiers were then constructed using the selected sequence features for predicting siRNA potency. Interestingly, gene expression inhibition is significantly affected by nucleotide dimer and trimer compositions of siRNA sequence.The findings in this study should help design potent siRNAs for functional genomics, and might also provide further insights into the molecular mechanism of RNA interference. RNA interference (RNAi) is a post-transcriptional gene regulatory mechanism by which a double-stranded RNA (dsRNA) induces sequence-specific gene silencing . The RNASince not all siRNAs are equally effective, siRNA design is one of the critical steps in gene silencing studies. Earlier experimental data have suggested several sets of empirical rules for designing potent siRNAs. For example, Ui-Tei et al.  proposedWith the accumulation of siRNA data, machine learning methods have been developed for both classification and regression analysis of siRNA potency. Strom  reportedThe above-mentioned previous studies suggest that many siRNA features of sequence composition, thermodynamic stability and secondary structure are related to the effectiveness of gene silencing. However, few advanced methods have been used to select the most relevant features for predicting siRNA potency. Most of the previous studies selected some siRNA features based on empirical knowledge or simple statistical analysis .More recently, Klingelhoefer et al.  used a sIn this study, random forests (RFs) were constructed to select important sequence features for predicting siRNA potency. RF-based variable importance measures were previously used in microarray expression data analyses to select a relatively small set of informative genes for disease/sample classification ,15. HoweAs described in the previous study , a non-rOf many potential features, only a few may be relevant for siRNA classification. This study examined 120 sequence features belonging to six groups  are randomly selected, and the tree node is split using the selected m variables. The random selection of features at each node decreases the correlation between the trees in the forest. Thus, the RF algorithm can handle many redundant features and avoid model overfitting. It has been shown that RFs outperform AdaBoost ensembles on noisy datasets, and can perform well on data with many weak input variables [In this study, important sequence features for siRNA classification were identified by using the random forest (RF) algorithm implemented in the software package available at ortance . Each deriables .x, its values in the oob instances associated with each tree in the forest are permuted randomly. The permuted oob instances as well as the original oob instances are then classified using the tree. The number of correct classifications on the original oob instances is subtracted by the number of predictions for the correct class on the permuted oob instances to calculate a raw score based on the tree. The importance score of variable x is defined as the average of raw scores over all the trees in the forest. For a fixed number of trees in the forest, the larger importance score a variable has, the more important it is for classification. In addition, a z-score can be obtained by dividing the variable importance score by its standard error, and a statistical significance level may be assigned to the z-score assuming normality [To evaluate the importance of variable rmality .SVMlight software package (available at http://svmlight.joachims.org/) was used to construct SVM classifiers. The SVM learning algorithm has been applied to a variety of biological problems for pattern classification, and may have superior generalization power with the ability to avoid overfitting [Support vector machines (SVMs) were trained with the selected sequence features to predict siRNA potency. The fitting . For a gfitting . The opt\u03b3 value makes the decision boundary smoother. Another parameter for SVM training is the regularization factor C, which controls the trade-off between low training error and large margin [\u03b3 and C parameters have been tested in this study to optimize the classifier performance.where  margin . DiffereWe used a fivefold cross-validation approach to evaluate the performance of SVM classifiers.Positive and negative instances were distributed randomly into five folds. In each of the five iterative steps, four of the five folds were used to train a classifier, and then the classifier was evaluated using the holdout fold (test data). The predictions made for the test instances in all the five iterations were combined and used to compute the following performance measures:TP is the number of true positives; TN is the number of true negatives; FP is the number of false positives; and FN is the number of false negatives. In addition to overall accuracy, sensitivity and specificity, Matthews correlation coefficient (MCC) is also commonly used as a measure of the quality of binary classifications [where cations . MCC mea against the false positive rate, which equals to (1 \u2013 specificity). In this work, the ROC curve has been generated by varying the output threshold of a classifier and plotting the true positive rate against false positive rate for each threshold value. The area under the ROC curve (AUC) can be used as a reliable measure of classifier performance [The Receiver Operating Characteristic (ROC) curve is probably the most robust approach for classifier evaluation and comparison . The ROCormance . Since tm parameter (the number of variables randomly selected to split each node in a tree). As suggested by the RF software [m . In this study, 10 RFs were constructed by varying the m parameter setting from 2 to 20 . Each RF with 1000 trees selected the top 20 features based on the z-score of variable importance. Some of the common features selected by the RFs were then identified for siRNA classification. The use of multiple RFs might increase the reliability for identifying relevant features.There are many potential features for siRNA classification. To select the important features, siRNA sequences were coded with the 120 features shown in Table oftware , the ceip = 0). Moreover, the frequency of UCC was found to be positively correlated with siRNA efficacy (Pearson's correlation coefficient r = 0.294). The frequency of UC was also selected as an important feature by the RFs . The other composition features positively correlated with siRNA efficacy include CG%, AAG%, AUC%, GCG%, AAC%, UUU%, ACA%, UUC%, and CAA% , whereas U% shows a positive correlation with gene expression inhibition (r = 0.127). Both the global G/C content and the G/C content in the first 5 bases (antisense strand) are negatively correlated with siRNA efficacy . In addition, the nucleotide identity at the third position  of the antisense strand may be an important feature for siRNA classification. A nucleotide C at the third position has a negative effect on siRNA efficacy (r = -0.199). However, the nucleotide identities at the other positions as well as siRNA secondary structure stability were not selected by most of the RFs.Several features of base composition and G/C content were also selected by the RFs. As shown in Table Some of the important features in Table \u03b3 = 1.2 and C = 1.2. As shown in Table To validate the important features selected by the RFs, support vector machines (SVMs) were trained with these features for siRNA classification. The siRNA instances were labeled with either 'potent' (positive instances showing \u226550% reduction in gene expression) or 'non-potent' (negative instances showing <50% reduction in gene expression). Classifier performance was evaluated using a fivefold cross-validation approach. Different settings of SVM training parameters were tested, and the best classifier was constructed with For performance comparison, SVM classifiers were also constructed using all the 120 features or only the 19 siRNA sequence features Table , suggestr = 0.4715). The result further suggests that the classifier RF_Features has learned some important siRNA patterns related to the efficacy of gene silencing.To further evaluate the classifier RF_Features, we examined the SVM output used to predict siRNA potency. The classifier was constructed using siRNA instances with binary labels (potent or non-potent). The actual magnitude of gene expression inhibition was not included in the training data. In Figure The classifier RF_Features compared favorably with several existing models. In the previous study by S trom , a similWe have developed a new machine learning approach for predicting siRNA potency based on random forests and support vector machines. Since there were many potential features for siRNA classification, random forests were used for feature selection based on variable importance scores. Interestingly, most of the selected features were nucleotide dimer and trimer compositions of siRNA sequence. Some nucleotide motifs  showed positive correlation with siRNA efficacy, whereas other motifs  might have a negative effect on gene silencing. These important features were used to train support vector machines for predicting siRNA potency with relatively high accuracy. In the future, we will apply our approach to a large, integrated dataset of siRNAs, and develop a software system for rational siRNA design in functional geneomic studies.The authors declare that they have no competing interests.LW initiated and designed the study. LW and CH conducted the data analysis. LW drafted the manuscript. JYY participated in experiment design, result interpretation and manuscript preparation."}
+{"text": "Intensive treatment for newborns with trisomy 13 is controversial because of their lethal prognosis. We report the better life prognosis of patients with trisomy 13 who received intensive treatment. At our hospital, we provided an intensive management to such patients including resuscitation and surgical procedures as required. Herein, we present the results of a retrospective study (1989\u20132010) of 16 trisomy 13 cases who received an intensive treatment. None was diagnosed to have trisomy 13 before birth; 9 were delivered by C-section and oxygen was administered to all patients during postpartum resuscitation. Mechanical ventilation was used in 9 patients after tracheal intubation and tracheotomy was performed in 2 patients when withdrawing of extubation was difficult. Regarding prognosis, 9 patients died, 3 were referred to another hospital, and 4 were discharged from the hospital. Four and 7 patients died within 7 and 30\u00a0days after birth, respectively. Nine patients survived for >1\u00a0month, 7 for >180\u00a0days, and 5 for >3\u00a0years. Median survival for 16 patients was 733\u00a0days. The patients who received intensive treatments survived longer compared to the previous data. This study provides useful information concerning genetic counseling, especially from an ethical point of view, before providing intensive management to newborns with trisomy 13. The . , 5. The . . Jones dThe approach to the management policy of a third-trimester fetus and infant with trisomy 13 and 18 is quite complicated, and the existing literature is controversial. The principal reason for the complexity surrounding decision making in the care of infants with trisomy 13 relates to high neonatal and infant mortality in both conditions , 8. AvoiIn Japan, the policies for the treatment of chromosomal anomalies including trisomy 13 and 18 with poor long-term prognoses are determined based on either of two controversial concepts: (i) the provision of thorough love and care while avoiding excessive intensive treatment; or (ii) the provision of active intensive treatment including resuscitation and surgery according to the clinical conditions of an infant and in accordance with the wishes of the infant\u2019s parents , 12. TheThe NICU at the Dokkyo Medical University Hospital provides the intensive treatments including resuscitation of intratracheal intubation, respiratory support, and some kind of surgery for trisomy 13 and 18, if informed consent is obtained from the family after counseling. At NICU, the five tentative treatment policies for trisomy 13 and 18 with poor long-term prognoses of survival are as follows: (1) The treatment policy is determined without relation with trisomy 13 and 18, and is determined after the pathology, treatment options and risks involved have been explained to the parents; (2) The actual initial treatment with resuscitation and medical therapy after delivery are provided to neonates who have ordinary cases, and surgical treatment is also not limited; (3) With respect to surgical treatment and resuscitation, even in case of long-term survival; (4) Psychological care for family is regularly provided by clinical psychotherapists; and (5) Management at home is actively discussed if the family wishes for it.In this study, in order to determine the survival period of trisomy 13 patients who received intensive treatment, we retrospectively analyzed the detailed clinical data of 16 patients with trisomy 13, admitted to NICU at the Dokkyo Medical University Hospital from 1989 to 2010.This study comprised 6,230 newborn infants who were hospitalized at the neonatal care unit of Dokkyo University Hospital, Japan during the period of 22\u00a0years from 1989 to 2010. Newborn infants presenting with external malformations or organ malformations that were observed during the clinical examinations underwent chromosomal analysis, and 183 patients (2.94\u00a0%) were diagnosed with chromosomal anomalies. Of these, 138 patients (2.22\u00a0%) were diagnosed with numerical autosomal aberration. A breakdown of these patients of numerical autosomal aberration shows trisomy 21 in 83 patients (1.33\u00a0%), trisomy 18 in 39 patients (0.63\u00a0%), and trisomy 13 in 16 patients (0.26\u00a0%). Other chromosomal abnormalities were 45 patients (0.72\u00a0%) including 2 patients with trisomy 8 mosaic and 2 patients with 4p-syndrome and others. In this study, we retrospectively investigated the NICU medical records at our hospital regarding the clinical details of 16 patients with trisomy 13 who received intensive treatment.None of the 16 patients with trisomy 13 had chromosomal abnormalities in their family histories. The age of the mothers ranged from 20 to 42\u00a0years (average: 32.9\u00a0years) including mothers who were over 35-years-old for 7 patients and over 40-years-old for 2 patients. The age of the fathers ranged from 22 to 47\u00a0years (average: 33.8\u00a0years) including fathers who were over 35-years-old for 7 patients and over 40-years-old for 3 patients. Regarding the history of previous delivery, 5 patients had parous mothers. In terms of gender, there were 10 males and 6 females. A definitive prenatal diagnosis of amniotic fluid test for trisomy 13 was not carried out for any of the 16 patients. However, in 15 patients diagnosed with trisomy 13, some type of abnormality was indicated in the ultrasonography test during the course of pregnancy, such as intrauterine growth retardation in 11 patients and brain malformations in 5 patients. The term of delivery ranged from 33 to 40 gestational weeks . Regarding the method of delivery, 9 patients were delivered via a cesarean section and 7 patients were delivered via a spontaneous cephalic delivery. The delivery term for the performance of cesarean section ranged from 33 to 40 gestational weeks .The average of Apgar Score at the time of birth was 4.25 points for 1\u00a0min scores and 6.5 points for 5\u00a0min scores. The birth weight ranged from 1,582 to 3,378 g, with an average weight of 2,243\u00a0g. The breakdown includes 7 patients weighing 1,500 to 2,000\u00a0g, 5 patients weighing 2,000 to 2,500\u00a0g, 2 patients weighing 2,500 to 3,000\u00a0g, and 2 patients weighting at least 3,000\u00a0g.The chromosomal karyotypes of all 16 patients were diagnosed using a G-band method. The r karyotypes included full trisomy 13 in 14 patients, mosaic type of trisomy 13 in 1 patient, and Robertsonian type of translocation in 1 patient.The major external craniofacial malformations included scalp defects in 5 patients for the head and cleft lip and palate in 10 patients for the face; malformation of abdomen included umbilical hernia in 8 patients. Polydactyly was noted in 6 patients for the extremities. Among the major organ malformations, 3 patients had alobar and 1 patient had semi-lobar type of holoprosencephaly. Dandy-Walker malformation was observed 1 patient and olfactory aplasia and fenestration of the septum pellucidum in 1 patient for the brain. As for the heart, some type of congenital cardiac disorder was observed in all patients. The breakdown includes atrial septal defect in 7 patients, ventricular septal defect in 6 patients, patent ductus arteriosus in 7 patients, coarctation of the aorta in 3 patients, endocardial cushion defect in 2 patients, tetralogy of Fallot (TOF) in 3 patients, and double-outlet right ventricle (DORV) in 3 patients. Combined respiratory malformations included congenital hypoplastic lung in 2 cases and congenital tracheal stenosis in 2 cases.Signs of acute respiratory failure were observed in 13 patients after birth. To resuscitate, oxygen was administered to all 16 patients. Mechanical ventilators were used for 9 patients to control acute respiratory failure after tracheal intubation. The tracheal intubation was unsuccessful in 1 patient due to severe tracheal stenosis.The various types of congenital malformations were observed in all 16 patients with trisomy 13; consultations with specialized surgeons were conducted to determine whether surgical treatment would be possible. Due to the difficulty of tube withdrawal after prolonged intubation, tracheotomy was performed for 2 patients. After tracheotomy, respiratory conditions in both patients were stabilized and they were able to survive for long-time period over 7\u00a0years. Abdominal operation for umbilical hernia was performed in 1 patient and for inguinal hernia in 1 patient. Plastic operation of the cleft lip and palate was performed in 1 patient in order to improve the cosmetic problem at 210\u00a0days of birth before discharge. Cardiac surgery was performed in 2 patients with trisomy 13. Ligation for patent ductus arteriosus and subclavian methods for coarctation of the aorta were carried out for 1 patient but she was died at 592\u00a0days of birth because of respiratory failure. Cardiac operation using Blalock\u2013Taussig method was performed for TOF in 1 patient at 330\u00a0days of birth and she has survived for more than 3\u00a0years and 3\u00a0months now.In-hospital death at NICU occurred in case of 9 patients, whereas 7 patients were discharged with the condition of homecare. Among them, 3 patients were referred by our hospital to another specialized institution close to the patients\u2019 homes for training purpose of homecare nursing at days 349, 251, and 63 after birth. The ages of 4 patients discharged from our hospital to their homes while they were alive were 336, 331, 256, and 204\u00a0days after birth. The average of median hospital stay regarding 7 patients who could be discharged including stay in another specialized institute was 256\u00a0days after birth. The survival period of the16 patients with trisomy 13 ranged from death at day 1 (within 24\u00a0h of birth) to 10\u00a0years and 1\u00a0month (this patient lives until now).The breakdown includes death at 1st day of birth, within 24\u00a0h, for 1 patient (6.25\u00a0%), death within 7\u00a0days of birth for 4 patients (25.00\u00a0%), and death within 1\u00a0month of birth for 7 patients (43.75\u00a0%). On the other hand, 9 patients survived for more than 1\u00a0month (56.25\u00a0%), 6 patients survived for more than 1\u00a0year (37.50\u00a0%), and 3 patients survived for more than 5\u00a0years (18.75\u00a0%). Therefore, the survival rates at ages of 1\u00a0day, 1\u00a0week, 1\u00a0month, 6\u00a0months, 1\u00a0year, 3\u00a0years and 5\u00a0years were 93.75, 75.00, 56.25, 43.75, 37.50, 31.25, and 18.75\u00a0%, respectively. Median survival time for all 16 patients, both males and females, were 733, 887, and 534\u00a0days, respectively.We also reviewed the data in detail to analyze the main cause of death. Some characteristic trend was recognized as follows: death occurred in 4 patients within 1\u00a0week of birth due to a major organ anomaly; especially, 3 patients had DORV and in 2 of 3, intubation with ventilation management was performed. On the other hand, 4 of 6 patients from long-survival group  had some kind of surgery that included tracheotomy in 2 patients and cardiac surgery in other 2 patients. Moreover, 5 of 6 patients from long-survival group (over 365\u00a0days) had a history of ventilation therapy.The longest survival time was set at 365\u00a0days Fig.\u00a0 and 1,82Among autosomal chromosomal abnormalities, birth is generally believed to be possible in patients of trisomies 21, 18, and 13. Of these, trisomies 18 and 13 are also referred to as fatal chromosomal abnormalities that lead to the development of many severe malformations . Most paAs for the prognoses of trisomy 13, there is one known report published in 2003 by the American Research Group regarding a large-scale survey of 5,515 cases. According to this report, the average age at the time of death was 10\u00a0days, but 5.6\u00a0% of the cases survived for more than 1\u00a0year . BesidesOf note, two reports have compared between active intensive treatment and non-active treatment with regard to prognosis of trisomy 18 , 14 but In conclusion, although our clinical data represent a small number of trisomy 13 patients, we did observe significantly better survival prognosis in the patients who received intensive treatment management. Our clinical results underscore the need to consider obtaining informed consent and counseling from patients\u2019 families for the future determinations of treatment policy for trisomy 13 patients."}
+{"text": "WeighsB mRNAs ; these gsB mRNAs . We haves Erk1/2 . HoweverWe have now defined the events leading from shear stress activation of NO/cGMP/PKG II to the activation of Src ; we showOur findings reveal crosstalk between NO/cGMP/PKG and integrins, establish a new mechanism of Src activation, and fill a gap in our understanding of how mechanical forces acting on cell-matrix adhesions are translated into cellular responses. Since Src controls Erk, a key regulator of osteoblast growth and survival, our results support using PKG-activating drugs as mechano-mimetics for treating osteoporosis."}
+{"text": "Despite the high prevalence of hearing impairment in the elderly, the rate of hearing aid use is still low. The objectives of this study were to report the nation-wide prevalence of hearing aid use in the Korean population and to determine the associated factors with hearing aid use utilizing a nationally representative data set.We obtained data from the 2010 to 2012 Korea National Health and Nutrition Examination Surveys, which were cross-sectional surveys of the civilian, noninstitutionalized population of the Republic of Korea at age \u226540 years . A field survey team performed interviews as well as physical examinations. Hearing aid use was assessed using an interviewer-administered questionnaire and pure-tone audiometry was administered for all participants in a sound-attenuating booth. Prevalence of hearing aid acquisition and regular use were calculated in participants who reported perceived hearing loss and who have bilateral hearing thresholds exceeding the 40\u200adB hearing level. Multivariable analyses were used to examine the associated factors with hearing aid use.The prevalence of hearing aid acquisition and regular use was 17.4% and 12.6%, respectively, in South Korea. Increased hearing threshold , severe perceived hearing loss , annoying tinnitus , balance problems , and myopia  were associated factors of regular use of hearing aids.The prevalence of hearing aid use in Korea is relatively low. Finding relevant factors of hearing aid use could provide further insight in setting up hearing-rehabilitation strategy for the elderly with significant hearing loss. Although hearing loss is highly prevalent in elderly adults and most can be successfully rehabilitated by using appropriate hearing aids, the uptake and use of hearing aids is still poor.4 Several studies have reported that a large proportion of people who could benefit from hearing aids do not wear them.6 The reported prevalence of hearing aid use among those with a hearing loss is low, ranging from 14.2% to 33.1%  was started in 1998 to examine the general health and nutrition status of population in South Korea. From 2010 to 2012, 10,000 to 12,000 individuals were selected annually and interviewed concerning their general health and nutrition, as well as undergoing a basic health assessment. In the otolaryngologic interview and examination, histories of tinnitus, hearing loss, and noise exposure were surveyed using structured questionnaires and endoscopic evaluations of the tympanic membrane as well as objective evaluation of hearing and balance were conducted in participants of appropriate ages.The present study was undertaken to determine the national prevalence of hearing aid use among participants >40 years of age with perceived hearing loss in South Korea based on survey data obtained from the 2010 to 2012 KNHANES and to investigate factors associated with hearing aid use.KNHANES is a national cross-sectional survey of the civilian noninstitutionalized population of South Korea. Korean Society of Otorhinolaryngology\u2013Head and Neck Surgery and Korean Ophthalmological Society support the survey. A field survey team that included an otolaryngologist, an ophthalmologist, and nurse examiners for health assessment moved with a mobile examination unit and performed interviews, physical examinations, and laboratory testing, including blood and urine collection, a pulmonary function test, a dental examination, an ophthalmologic examination, and an otolaryngologic examination. Every year, 7000 to 10,000 individuals in \u223c3500 households are selected from a panel to represent the population by using a multistage clustered and a stratified random sampling method that is based on the National Census Data. The 500 survey areas were drawn from a census of population and housing by considering the proportion of each subgroup. The participation rate of selected households in the past several cycles of KNHANES has been high, \u223c80%. A total of 23,621 individuals, representing the 47,761,045 individuals in South Korea, participated in the survey from 2010 to 2012, and 12,709 individuals >40 years of age were initially included in this study. Written informed consent was obtained from all participants before the survey, and approval for this research was obtained from the Institutional Review Board of the Samsung Medical Center (IRB No. 2013-02-031).10 To evaluate the experience of balance problem, participants were asked whether they had experienced dizziness or imbalance, positional dizziness, and falls in the absence of external forces in the past 12 months.Participants were first asked about their perceived hearing loss. In detail, the participants were asked to rate their difficulty in hearing (\u201cWhich among the below best describes your [unaided] hearing?\u201d) by identifying their hearing as having no difficulty, mild difficulty, significant difficulty, or as no hearing. The respondents who professed any degree of hearing difficulty were then asked about their use of hearing aids. Responses to the question \u201cDo you use hearing aid(s)?\u201d were \u201cyes\u201d, \u201cyes, but rarely use it\u201d, or \u201cno\u201d. The participants were considered to regularly use hearing aids only if they answered \u201cyes\u201d. To determine the prevalence of tympanic membrane perforation and cholesteatomatous conditions, including retraction pocket and otitis media with effusion, an ear examination was conducted with a 4-mm 0\u00b0-angled rigid endoscope attached to a CCD camera for all participants. The pure tone air-conduction threshold was measured in a sound-proof booth using a model GSI SA-203 automatic audiometer . Moderate-to-profound hearing loss was defined as > 40\u200adB hearing level with average air-conduction hearing thresholds measured at 0.5, 1, 2, and 3\u200akHz according to the recommendation from the American Academy of Otolaryngology\u2014Head and Neck Surgery.P\u200a<\u200a0.20) in the univariable analysis were entered in the multivariable analysis model. To reflect national population estimates, sample weights were applied in all analyses. P values and 95% CI were corrected by Bonferroni's method in the case of multiple testing. In all tests, P values were 2 sided, and P\u200a<\u200a0.05 was considered to be statistically significant. Statistical analysis was performed using SAS 9.3 .The prevalence and 95% confidence interval (CI) for hearing aid use were calculated. To examine the association of various covariates on the use of hearing aids, univariable analysis and multivariable analysis were performed using the logistic regression model. Biologically and medically plausible covariates, as well as the variables found to be possibly associated . Among these, those who did not respond to the questionnaire queries of hearing aid use, tinnitus, noise exposure, and perceived hearing loss, or those who did not underwent otolaryngologic or ophthalmologic examinations were excluded. Additionally, those who had no perceived hearing loss (who responded to have \u201cno difficulty\u201d in subjective hearing) were also excluded. Finally, 530 participants, representing 732,618 individuals with objective bilateral moderate-to-profound hearing loss (\u2265 40\u200adB) and perceived they had hearing loss, were included in this study. Among the 530 participants, 94  answered that they had hearing aids. However, 25 participants  rarely used hearing aids even though they had them. Thus, the remaining 69 participants  who regularly used hearing aids in daily life were considered as hearing-aid user; the prevalence of hearing aid use was 12.6% (95% CI 9.0%\u201316.2%).Hearing aid use in participants >40 years of age with bilateral moderate-to-profound hearing loss and associated factors were investigated using univariable and multivariable analyses. Table P\u200a<\u200a0.20) in the univariable analysis were entered in the multivariable analysis model. Multivariable logistic regression analysis revealed that high hearing threshold  and number of house members  were significant associated factors of regular hearing aid use among hearing aid owners.Among 94 participants who had hearing aids, only 69 participants regularly used their hearing aids and the remaining 25 participants were nonusers. Thus, in order to identify factors associated with regular hearing aid use in all hearing aid owners, additional univariable and multivariable analyses were performed. Table 11 Besides its high incidence and prevalence, age-related hearing impairment affects quality of life enormously by reducing communicative relationships, as well as reducing social and emotional interactions.12 Thus, appropriate tools for hearing rehabilitation are needed, and hearing aids are recommended for majority of people with age-related hearing impairment. Previous studies have reported that hearing aids can improve the ability to communicate, improve quality of life, prevent social isolation, and positively affect hearing-related depression.16 In addition, by modeling different degrees of hearing loss with a multistate model, hearing aid use was proven to be a cost-effective strategy to rehabilitate the hearing-impaired elderly.17 However, many people with impaired hearing do not wear them. Over the last 20 years, hearing aid adoption has remained at \u223c 20% with an admitted hearing loss.8These are the first national estimates of hearing aid prevalence in the Korean population based on audiometric data and a large, government-centered, well-selected representative sample. Age-related hearing impairment is a hallmark of aging; therefore, the prevalence and severity increase with age.20 In European countries, such as the United Kingdom, France, Denmark, and the Netherlands, expenses of hearing aids are covered by public health insurance. In the United States, the use of hearing aids has continually increased and is now \u223c23%, with most of this growth attributed to free hearing aids obtained through the Department of Veterans Affairs or low-cost Internet sales.8 However, in South Korea, the expense of hearing aids is not covered by the government health insurance system, and the average price of hearing aids is relatively high when considering the national income per capita. In addition to cost, cultural attitudes that undervalue the role of hearing on life as well as stigma associated with hearing aid use partly account for the low rate of hearing aid use in South Korea.21In this study, national prevalence of hearing aid use in South Korea was 12.6% among people with moderate-to-profound hearing loss, which is relatively low as compared with those reported in other countries  than those who declined to use the aids .23At present, both objective hearing thresholds and subjective assessment of participant's own hearing loss were associated with hearing aid use. People who use hearing aids have more severe hearing losses than nonusers. Considering the odds ratio of both objective and subjective hearing loss (1.05 vs 10.73), self-reported auditory difficulty is more important in using hearing aids than objective hearing sensitivity. Perceived hearing handicap assessed by the Hearing Handicap Inventory for the Elderly questionnaire is associated with the hearing aid use, with an adjusted odds ratio for a 1-unit increase in HHIE scores of 1.08.24 The use of hearing aids in tinnitus makes the patient less aware of the tinnitus and reduces the annoying sensation that sounds and voices are masked by the tinnitus, by amplifying external sounds and subsequent activation of the auditory nervous system.26 Survey and case control studies have shown that tinnitus patients gain relief from the symptoms by the consistent use of hearing aids.28 In addition, a broader range of patients with milder degrees of hearing loss can become successful hearing aid users due to the increased use of open-fit hearing aid. This can benefit tinnitus sufferers who previously were not interested in amplification.Patients with bothersome tinnitus had an increased tendency to use hearing aids in this study. Hearing aids are often recommended as a treatment option for patients who suffer from bothersome tinnitus and who have moderate to severe hearing loss. This practice is supported by recently published clinical practice guideline of tinnitus, recommending a hearing aid evaluation for patients who have persistent, bothersome tinnitus associated with documented hearing loss.29 Recently, it was shown that hearing aids are a novel treatment modality for imbalance in older adults with hearing loss.30 That is partly attributed to importance of auditory input in maintaining balance, because audition may represent as an independent sensory input providing spatial sound cues. However, as reported in this study, the rate of hearing aid use among people with balance problem is low, probably because of difficulties in purchasing and managing hearing aids due to restricted activity or inadequate awareness about the beneficial effect of hearing aids on balance. Balance deficits are a well-known risk factor for falls in the elderly, and both balance deficits and subsequent fear of falling are associated with restricted physical activity.31 Therefore, raising public awareness about the benefits of hearing aid use and appropriate rehabilitation programs to increase physical activity would be helpful in this specific population who both has hearing impairment and balance problems.Presbycusis is associated with an increased risk of falls.32 A loss of visual ability can make it difficult for the elderly to insert a hearing aid into the ear canal, replace a battery, or provide hearing aid maintenance34 because poor vision can affect manual dexterity in aged population. Furthermore, many participants with both hearing and vision impairments express concerns about managing hearing aids due to their low vision,35 which may explain the low rate of hearing aid use in individuals with myopia. In addition, people with low vision often depend on spatial hearing and echolocation. However, if hearing loss is combined, then safe mobility may be compromised by loss of accuracy of sound localization and reduced perception of cues.36 Thus, active provision of hearing-rehabilitation services is required for this dual sensory impairment.Having myopia was associated with decreased use rate of hearing aids in this study. Vision and hearing impairments are common among the elderly and can occur separately or in combination.37 It is also important to identify comprehensive list of risk-factors for compliance with hearing aid use. The most important factors for regular hearing aid use (good compliance) in this study were the number of house member and hearing threshold. This result suggests that needs for communication with house (family) members can positively affect motivation of hearing-impaired listeners to use hearing aids regularly. It also indicates that hearing-impaired listeners with small family can be at higher risk for nonregular hearing aid use, and clinicians should pay attention to this specific population during the counseling and fitting of their aids.In this study, among 94 participants who purchased hearing aids, 25 participants (26.6%) rarely used hearing aids. This result is in line with prior research from UK, indicating that a total of 29% of hearing aid owners do not use their hearing aids on a regular basis.A possible limitation of this study is the lack of comprehensive inclusion of possible associated factors that include source of motivation, attitudes toward hearing aid, expectations of hearing aids, and individual experiences of hearing-aid counseling. However, the key attributes of this study lie in the national cross-sectional survey with representative population based on the National Census Data, providing detailed and standardized audiometric data, as well as in the comprehensive range of demographic and clinical variables accurately measured by an otolaryngologist, ophthalmologist, and nurse examiners. These data also enable a thorough evaluation of associated factors with hearing aid use among older (\u226540 years of age) adults in South Korea. The results could be used in devising strategies to counsel the hearing-impaired people and to increase hearing aid adoption rates.The prevalence of hearing aid use is estimated as 12.6% in South Korea. Increased hearing threshold, severe perceived hearing loss, accompanying annoying tinnitus, balance problems, and myopia are associated with hearing aid use. Among these factors, increased hearing threshold, severe perceived hearing loss, and accompanying annoying tinnitus may be easily expectable, but balance problems and myopia were newly found to be associated with hearing aid use. Therefore, concerns about balance disorder and myopia may need to be raised during hearing aid counseling. Additionally, hearing-impaired listeners with higher hearing thresholds and small family are at risk for nonregular hearing aid use.This information should be incorporated into daily clinical practice, and interventions to reduce barriers of hearing aid use seem to be necessary in satisfying an unmet need, and to reduce negative consequences from hearing impairment in the elderly. Further researches are needed to evaluate the effect of active/professional counseling and long-term follow-up program on regular hearing aid use."}
+{"text": "Hearing loss is associated with poor cognitive performance and incident dementia and may contribute to cognitive decline. Treating hearing loss with hearing aids may ameliorate cognitive decline. The purpose of this study was to test whether use of hearing aids was associated with better cognitive performance, and if this relationship was mediated via social isolation and/or depression. Structural equation modelling of associations between hearing loss, cognitive performance, social isolation, depression and hearing aid use was carried out with a subsample of the UK Biobank data set  of UK adults aged 40 to 69 years who completed a hearing test. Age, sex, general health and socioeconomic status were controlled for as potential confounders. Hearing aid use was associated with better cognition, independently of social isolation and depression. This finding was consistent with the hypothesis that hearing aids may improve cognitive performance, although if hearing aids do have a positive effect on cognition it is not likely to be via reduction of the adverse effects of hearing loss on social isolation or depression. We suggest that any positive effects of hearing aid use on cognition may be via improvement in audibility or associated increases in self-efficacy. Alternatively, positive associations between hearing aid use and cognition may be accounted for by more cognitively able people seeking and using hearing aids. Further research is required to determine the direction of association, if there is any direct causal relationship between hearing aid use and better cognition, and whether hearing aid use results in reduction in rates of cognitive decline measured longitudinally. The prevalence of dementia in those aged over 60 years is between 5\u20137%, with the numbers of those affected globally forecast to double every 20 years between 2010 and 2050 ,4, and tHearing loss is common in older adults  and is aThere is evidence that intervention in the form of hearing aids may improve quality of life and increase social engagement  and incoThe aim of this study was to model statistical associations between hearing impairment and cognitive performance in a large and inclusive data set. A positive association between hearing ability and cognitive performance could be consistent with both the cascade and common cause hypotheses. However, if auditory deprivation contributes to cognitive decline, as suggested by the cascade hypothesis, use of hearing aids should be associated with better cognitive performance. The mediating role of social isolation and/or depression was also investigated.http://www.ukbiobank.ac.uk/). All data were anonymized and de-identified prior to analysis. As UK Biobank data collection proceeded, additional measures were included for a subset of participants. Participants in the present study were a subset of 164,770 who were asked to complete a hearing test (the Digit Triplet Test).UK Biobank was established for prospective investigations of the genetic, environment and lifestyle causes of diseases of middle and older age . EthicalSex, ethnicity data (based on 2001 UK Census categories) and Townsend deprivation score (based on the area of residence) were collected for each participant. Townsend deprivation scores are widely used in health studies as a proxy for socioeconomic status . Lower Tr = 0.77) with audiometric thresholds [http://www.actiononhearingloss.org.uk/your-hearing/look-after-your-hearing/check-your-hearing/take-the-check.aspx). The DTT procedure is described elsewhere . Briefly, the signal to noise ratio (SNR), reported in decibels, for the 50% correct speech recognition threshold was estimated for each ear. The level of hearing loss was based on better ear performance. Hearing aid users performed the DTT without hearing aids.The Digit Triplet Test (DTT) is a speech-in-noise test originally developed in Dutch for reliable large scale hearing screening, and which correlates highly . Hearing loss would not be expected to contribute to performance on these visually presented tests. The background and rationale for the cognitive tests is reported by UK Biobank elsewhere (http://www.ukbiobank.ac.uk/wp-content/uploads/2011/11/UK-Biobank-Protocol.pdf?phpMyAdmin=trmKQlYdjjnQIgJ%2CfAzikMhEnx6).Cognitive tests were completed via a computerised touch screen interface. Further information is reported elsewhere  was based on multiple choice responses to 13 questions such as \u201c\"Bud is to Flower as Child is to?\" Participants had 2 minutes to complete as many questions as possible. Questions that were not completed within the time limit were scored as zero. The outcome measure was the sum of the number of correct answers.Hearing aid use was assessed via response to the question \u201cDo you use a hearing aid most of the time?\u201d Social isolation was assessed via response to the question \u201cDo you often feel lonely?\u201d Participants had the response options Yes/No/Do not know/Prefer not to answer. Depression was measured via response to the screening question; \u201cOver the past two weeks, how often have you felt down, depressed or hopeless?\u201d . ParticiStructural equation modelling  was usedwww.statmodel.com/). Fit statistics and standardised coefficients were reported for each model. The Mplus estimates for paths from predictors to an observed categorical dependent variable  are probit regression coefficients. A positive sign means that the probability of the categorical dependent variable (e.g. the category 1 for a 0/1 variable) is increased when the predictor value increases. A larger magnitude means that this probability is higher. For the standardised latent cognition variable a higher score implies worse cognition due to the direction of the factor loadings. The depression variable with four response levels was treated as a continuous variable. Estimates for paths from predictors to these dependent variables can be interpreted as in a standard linear regression.Cognition was measured by a standardised latent factor  in the structural equation model which was derived from a measurement model with observed indicators of the reaction time, pairs matching and fluid IQ tests. The covariates age, sex, general health  and socioeconomic status (Townsend index) were included as predictors for each outcome variable in the overall structural equation model. Modelling was carried out using robust weighted least squares (WLSMV) in the Mplus program version 7.11 (In Model 1, after controlling for age, sex, SES, and general health, poorer hearing remained significantly associated with poorer cognition . HoweverIn cross-sectional modelling in a large sample of UK adults, hearing aid use was associated with better cognition. This is consistent with the \u2018cascade hypothesis\u2019, where long-term auditory deprivation or degraded auditory input may result in increased cognitive decline ,12,13. TIf hearing aids do have a positive impact on cognitive performance not due to a reduction in depression or social isolation, how might hearing aid use impact on cognition? According to the cascade hypothesis, untreated hearing loss may result in long-term auditory deprivation or degraded auditory input, resulting in increased cognitive decline. However, the mechanism for this is not known and requires elucidation ,12,13. OOne unexpected result was a lack of association between hearing aid use and depression, and increased social isolation associated with hearing aid use. It may be that hearing aids do discourage participation in social events by amplifying aversive background noise that is typical at social venues such as clubs, cafes and restaurants. However, hearing aids have been previously suggested not only to reduce hearing handicap, but to reduce concomitant social isolation and depression . EvidencThe assumption in the present study was that better cognition in hearing aid users observed in cross-sectional analysis may reflect the long-term impact of hearing aid use in reducing cognitive decline. However, longitudinal data are required to confirm whether hearing aid use is associated with any alteration in the rate of cognitive decline over time. The data in the present study are correlational, and no strong conclusions about causality are possible. Alternative interpretations of the patterns of association reported in the present study are possible. For example, rather than hearing aids \u2018causing\u2019 better cognition, cognitively more able people might tend to obtain and use hearing aids. Cognitively more able people may be more likely to access specialist health services, including audiology, or may more likely recognise hearing disability and seek treatment. Establishing a causal association between hearing aid use and cognitive performance requires controlled studies with cognitive outcomes measured in the short term as well as after several years hearing aid use. The study was restricted to adults aged 40 to 69 years, so it is uncertain whether the associations identified in the present study are generalizable to older adults, in whom sensory impairment, hearing aid use and cognitive impairments are more common.Hearing aid use was associated with better cognition in a large cross-sectional study of UK adults. The association was independent of social isolation and depression. Further research is required to determine the direction of association, if there is any direct causal relationship between hearing aid use and better cognition, and whether hearing aid use results in reduction in rates of cognitive decline measured longitudinally. Treating hearing loss may make a significant contribution to reducing the burden associated with cognitive decline and reduced quality of life."}
+{"text": "Background. Previous research has found that hearing loss is associated with poorer cognitive function. The question is that when a hearing impairment is being compensated for by appropriately fitted monaural hearing aids, special precautions are still needed when screening cognitive function in older adults. Objective. This research examined cognitive function in elderly hearing aid users who used monaural hearing aids and whether the presence of a hearing impairment should be accounted for when screening cognitive function in these individuals. Methods. Auditory thresholds, sentence reception thresholds, and self-reported outcomes with hearing aids were measured in 34 older hearing aid users to ensure hearing aids were appropriately fitted. Mini-Mental State Examination (MMSE) results obtained in these participants were then compared to normative data obtained in a general older population exhibiting similar demographic characteristics. Stepwise multiple regression analyses were used to examine the effects of demographic and auditory variables on MMSE scores. Conclusions. Results showed that, even with appropriately fitted hearing aids, cognitive decline was significant. Besides the factors commonly measured in the literature, we believed that auditory deprivation was not being fully compensated for by hearing aids. Most importantly, screening of cognitive function should take into account the effects of hearing impairment, even when hearing devices have been appropriately fitted. Besides hearing impairment, decline in cognitive functions is also commonly observed in the aging population. Recent studies showed that reduced auditory input due to a hearing impairment is also associated with greater declines in cognitive function in older adults than those without hearing loss.Lin and colleagues  showed tVarious hypotheses have been proposed to explain this decline associated with hearing impairment. While there could be a common neuropathologic origin that underlies both hearing and cognitive decline, the hearing loss could possibly lead to a cycle of multimorbidity in different areas or may interact with other risk factors to accelerate cognitive declines . The hean = 13) with an average hearing loss of about 50\u2009dB HL. Using the MMSE, Acar et al. /4 . StepwisOverall, results from audiological assessment and the CHINT suggest that hearing aids brought significant benefits in terms of improving sensitivity to sounds and speech reception. Results from the IOI-HA also revealed that participants were using their hearing aids consistently and reported very positive outcomes with the hearing aids. In fact, patient records showed that their hearing aids were adjusted to the satisfaction of the users and further adjustment was not needed. Thus, we were ensured by these results that the hearing aids had been appropriately fitted.Results obtained on the MMSE showed slight but significant decline in overall cognitive function (with moderate to large effect size) among the subject population. Interestingly, out of the eight domains measured on the MMSE, auditory factors  predicted the scores on five MMSE domains that required understanding of the verbal instructions. Auditory or demographic variables did not predict scores measured on Attention and calculations, Recall, or Language. Nonetheless, these findings suggested that hearing and cognition are intricate aspects of the aging process.While the current study could not delineate which of the hypotheses mentioned above made greater contribution to the results, we would elaborate on three issues here. First, there was deprivation of auditory inputs in the participants. As mentioned above, the users waited an average of 11 years before they obtained hearing aids. The long-term deprivation in auditory inputs prior to hearing aid fitting might not be fully reversible even with the use of hearing aids. Similarly, monaural hearing aid use might have resulted in the unaided ear being deprived of auditory inputs. Although it would be difficult to control the duration of the wait to get intervention, future research could compare cognitive outcomes between monaural and binaural users.Second, one could argue that more gain could be provided to further optimize the hearing. Although the participants felt that their hearing aids were fitted appropriately, aided hearing thresholds obtained in the soundfield were improved to 42\u2009dB\u2009HL, which meant that some of the weaker signals were not audible. When the hearing disability is not being fully compensated for, the efforts spent on understanding conversations may result in fewer resources being left for storing information . ListeniFinally, we could not rule out the possibility that some participants might have difficulties understanding the verbal instructions. While there were several measures to ensure optimal understanding of the instructions during cognitive testing, it would be difficult to rule out the possibility that some instructions were not heard clearly. As discussed above, auditory factors seemed to influence scores on the MMSE domains that required understanding of the verbal instructions, although the effect sizes are only small to medium. Schneider and Pichora-Fuller  also fouThe present and previous researches have shown that hearing aid users tend to wait a long time before they take up hearing devices . As hearEmerging evidence is showing that hearing aid users with poorer cognitive function are less able to take advantage of more advanced signal processing algorithms that are supposed to aid speech understanding. These may include noise reduction and compression with short time constants and directional microphones \u201326. TherIn Hong Kong, it is uncommon for individuals with hearing impairment to receive aural rehabilitation, other than the fitting of amplification devices. Via a meta-analysis, Chisolm and Arnold  have shoWe reported findings from a preliminary study and they are somewhat limited by the small sample size. However, the results will help us plan follow-up studies that address the imminent issues. A larger scale study is being carried out in our laboratory to examine the application of several cognitive tests in the subject population. The current study took a cross-sectional view of cognitive function in a general hearing aid user population; a carefully planned longitudinal study would hopefully help establish a causal relationship between the long-term use of hearing aids that are appropriately fitted and cognitive function. Learning about whether hearing aid use could reverse or arrest the progression of cognitive decline is essential for clinicians to make evidenced based recommendation on early hearing aid use. It will also be interesting to find out whether other invention options, such as the use of binaural hearing aids and perceptual and cognitive training, could improve cognitive function to a level commensurate with that of the general older population.Overall, the present study showed that while appropriately fitted monaural hearing aids could partially make up for the hearing disability and improve speech understanding, the use of hearing aids may not fully compensate for the decline in cognitive function associated with hearing loss. Therefore, when screening cognitive function, the presence of a hearing impairment should be accounted for. In particular, ensuring audibility of signals and perhaps the use of cognitive function tests that employ visual presentation of stimuli should be used. We have also identified a few research areas where greater understanding on the relationship between cognition and hearing impairment would improve the clinical management of older patients."}
+{"text": "In many countries including the United Kingdom, hearing aids are a first line of audiologic intervention for many people with tinnitus and aidable hearing loss. Nevertheless, there is a lack of high quality evidence to support that they are of benefit for tinnitus, and wide variability in their use in clinical practice especially for people with mild hearing loss. The aim of this study was to identify a consensus among a sample of UK clinicians on the criteria for hearing aid candidature and clinical practice in fitting hearing aids specifically for mild hearing loss with and without tinnitus. This will allow professionals to establish clinical benchmarks and to gauge their practice with that used elsewhere.The Delphi technique, a systematic methodology that seeks consensus amongst experts through consultation using a series of iterative questionnaires, was used. A three-round Delphi survey explored clinical consensus among a panel of 29 UK hearing professionals. The authors measured panel agreement on 115 statements covering: (i) general factors affecting the decision to fit hearing aids, (ii) protocol-driven factors affecting the decision to fit hearing aids, (iii) general practice, and (iv) clinical observations. Consensus was defined as a priori \u226570% agreement across the panel.Consensus was reached for 58 of the 115 statements. The broad areas of consensus were around factors important to consider when fitting hearing aids; hearing aid technology/features offered; and important clinical assessment to verify hearing aid fit (agreement of 70% or more). For patients with mild hearing loss, the greatest priority was given by clinicians to patient-centered criteria for fitting hearing aids: hearing difficulties, motivation to wear hearing aids, and impact of hearing loss on quality of life (chosen as top five by at least 64% of panelists). Objective measures were given a lower priority: degree of hearing loss and shape of the audiogram . Areas where consensus was not reached were related to the use of questionnaires to predict and verify hearing aid benefit for both hearing and tinnitus; audiometric criteria for fitting hearing aids; and safety of using loud sounds when verifying hearing aid fitting when the patient has tinnitus (agreement of <70%).The authors identified practices that are considered important when recommending or fitting hearing aid for a patient with tinnitus. More importantly perhaps, they identified practical issues where there are divided opinions. Their findings inform the design of clinical trials and open up debate on the potential impact of practice differences on patient outcomes. This study identified areas of consensus among 29 United Kingdom hearing professionals on the criteria for hearing aid candidature and clinical practice in fitting devices for mild hearing loss with and without tinnitus. Consensus was strong for factors important to consider when fitting hearing aids, hearing aid technology/features offered, and important clinical assessment to verify hearing aid fit. The results will inform the design of clinical trials and open up debate on the potential impact of practice differences on patient outcomes. Tinnitus is a major problem affecting 10 to 15% of adults depending on the study , with abIn the United Kingdom, audiology is the main provider of care for people with tinnitus  and the UK  guidelinThere have been a number of tinnitus-specific management guidelines. Few of them provide specific recommendations of best practice  among the panel. The primary aim of our study was to identify clinical consensus on the criteria for hearing aid candidature and practices for fitting hearing aids for mild hearing loss with and without tinnitus to inform the design of clinical trials. We did not wish to draw any conclusions about the effectiveness of current practice or formulate any clinical recommendations or guidelines at this stage. Therefore, a consensus of \u226570% agreement was considered appropriate to interpret the data. In addition, Kendall\u2019s coefficient of concordance W was calculated to assess the degree of agreement between panelists in the final round of the questionnaire (round 3). Statistical analysis was performed in R 3.0.2 . MultiplThe study design followed a standard Delphi survey methodology  where thOur Delphi protocol consisted of five stages.Our long-term goal is to design and conduct a gold standard RCT to evaluate the efficacy of hearing aid interventions for people with tinnitus. A key step in this process is to document and better understand current practices for fitting hearing aids to ascertain the feasibility of conducting such a study. The immediate goal of the current study was to identify consensus to establish clinical benchmarks and to enable clinical professionals to gauge their practice with that elsewhere.An invitation letter was sent to 65 UK-based audiology and hearing therapy staff who had responded to a previous survey of tinnitus management . The letForty-two clinicians from the National Health Service (NHS) and independent audiology centers agreed to participate in the study. Twenty-nine of the 42 clinicians responded to round 1 (69% response rate), and they formed our expert panel We asked panelists to list up to five audiologic and nonaudiologic factors that they consider important when fitting hearing aids for patient with mild hearing loss and to identify differences in practice when fitting hearing aid(s) to a patients who have bothersome tinnitus as a main complaint.ii.Case studies (seven questions)http://links.lww.com/EANDH/A175). We asked \u201cAssuming that the patient is motivated to follow your recommendation, under what circumstances and why would you fit a hearing aid to someone with these hearing profiles?\u201d Panelists were asked to identify up to three reasons they would fit a hearing aid to patients with such an audiogram. They were asked to consider patients with and without bothersome tinnitus separately.We presented panelists with seven audiograms, which varied in shape and the degree of hearing loss Panelists were asked to identify hearing aid technology (devices/products) offered, prescription formulae, and assessments they performed to verify hearing aid(s) fitting for patients with mild hearing loss with and without tinnitus. We also asked about any departmental limitations or protocols, and what panelists would do differently if there were no constraints on what they could offer patients.iv.CommentsPanelists were asked for any additional comments relating to this round of the survey.To identify themes and to formulate closed questions for round 2, responses to open questions from round 1 were subjected to thematic analysis . ThematiThe round 2 questionnaire consisted of 115 statements grouped into four themes derived from the thematic analysis process: (i) general factors affecting the decision to fit hearing aid(s); (ii) protocol-driven factors affecting the decision to fit hearing aid(s); (iii) general practice; and (iv) clinical observations. These themes, based on information provided by the panel, rather than predefined categories created by researchers in round 1, are used later to describe the results.Each statement in the questionnaire was attached to a 5-point Likert rating scale  for panelists to quantify their level of agreement with each statement. For question 1, panelists were also asked to choose the five most important factors from the list of 20 that affect the decision to fit hearing aids and rank them in the order of importance (1 = first most important to 5 = fifth most important).The round 3 questionnaire consisted of the same list of closed questions used in round 2. For each item, the summary results from round 2 were reported back to the panelists. Results were anonymized, and the mean level of agreement for each of the closed questions was given. Considering this information, panelists were given the opportunity to revise their personal rating of any statement. A comment box was introduced for 20 statements in which low agreement had been observed in round 2. The comment box invited panelists to explain their round 3 rating. Results from the round 3 questionnaire were taken as the final level of agreement.The three-round Delphi survey was conducted between October 2012 and June 2013 (Table Consensus (\u226570% agreement) was reached for 58 of 115 statements, with an average agreement of 89.9% (SD = 9.9%) for these statements Table . KendallThe number of statements for which the consensus was reached increased from 50 to 58 between rounds 2 and 3. The percentage agreement between rounds 2 and 3 increased for 57 of 115 statements, decreased for 14, and stayed the same for 44 statements. The increase in percentage agreement for 57 statements varied from 1 to 17% , and the decrease for 14 statements varied from 0.5 to 11% . In most cases, the decreases in agreement concerned statements that already had a low level of agreement (<67%) after round 2.Round 3 identified where there was a panel agreement for the 20 factors that clinicians reported in round 1 to be relevant to the decision about fitting hearing aids for the patient with mild hearing loss but without tinnitus. Greater priority was given by clinicians to patient-centered criteria for fitting hearing aids than the more objective assessment measures.Consensus was reached for 11 of 20 factors. Thematic analysis of the panelists\u2019 answers pointed to the importance of two groups of factors when deciding hearing aids candidature for mild hearing loss: patient-centered criteria and objective measures. Patient-centered criteria included patient-reported hearing difficulties, motivation to wear hearing aid(s), realistic expectations, patient self-reported impact of hearing loss on quality of life, general ability to hear in quiet settings and in background noise, and speech discrimination and comprehension in quiet and background noise. Clinicians agreed that among the objective measures, shape of the audiogram, degree of hearing loss, health of the ears, manual dexterity and manipulation difficulties, and hyperacusis/reduced loud sound tolerance all play an important role when deciding whether to fit hearing aids for a mild hearing loss Table .Patient-centered criteria were given higher importance than objective measures by our panelists Table . Twenty-Factors for which consensus was not reached are listed in Table In summary, the thematic analysis of round 1 responses identified 14 statements concerning protocol-driven factors affecting the decision to fit hearing aid(s), which formed this part of the survey. While consensus was reached on the opinion that hearing aids are the main intervention for patients with mild hearing loss with tinnitus, there was no consensus on the criteria for determining the candidature both objective (audiometry) and self-reported (questionnaires).Consensus was reached for 7 of 14 statements. Panelists agreed that hearing aids should be offered routinely for patients with mild hearing loss and bothersome tinnitus (89%) and that hearing aids are the primary treatment for mild hearing loss with bothersome tinnitus (72%).On fitting, panel members reached consensus that unilateral hearing aid fitting in patients with mild hearing loss and bothersome tinnitus is not appropriate even if the patient has unilateral tinnitus or an asymmetric hearing loss  only if patient had a four-frequency pure-tone average worse than 20 dB at least in one ear (75% agreement). They disagreed with the statement that hearing aids should be offered only if the patient showed \u226535 dB hearing loss at 2\u2009kHz . These included high-frequency hearing loss (>6\u2009kHz), hearing loss >20 dB for at least two octave frequencies, and affected frequencies being within speech range.Consensus was not reached for statements on the use of questionnaire scores as a criterion for fitting hearing aid(s)  for patients with mild hearing loss with and without tinnitus included in this section. Panelists reached consensus for a wide range of statements about candidature for hearing aids, recommended technology, and verification of hearing aids\u2019 fit.Consensus was reached for 23 of 48 statements, which we categorized as (i) candidature for hearing aid(s), (ii) hearing aid technology/features, and (iii) verification of hearing aid(s) fit.i. Candidature for hearing aid(s)All panelists agreed (100%) that they would always provide information about the potential benefit of hearing aids for tinnitus management. There was consensus that hearing aid(s) should be offered for patients with a mild hearing loss and bothersome tinnitus even if they did not report hearing difficulties (82%). Panelists also agreed that they were more likely to fit hearing aid(s) for mild hearing loss if the patient also had bothersome tinnitus (86%). Ninety-three percent agreed that they would offer hearing aid(s) to a patient with a mild hearing loss and bothersome tinnitus if the patient did not want or could not use a noise generator.ii. Hearing aid technology/featuresConsensus was also reached around hearing aid technology/features offered. Generally for mild hearing loss, clinicians would use open fit (100%), behind the ear or on the ear devices (100%) with multiple programs (93%), directional microphone (96%), and slim/thin tubing (93%). For patients who also have bothersome tinnitus, panelists would especially focus on providing as open fit as possible (100%). For someone with tinnitus, they would also aim to reduce feedback (93%), use slim/thin tubing (93%), use separate programs for optimizing speech and audibility (71%), and use frequency compression for high-frequency hearing loss, that is defined as mild loss according to the 4-point  pure-tone average (71%).iii. Verification of hearing aid(s) fitObjective  and subjective measures  were identified as important to verify the fit Table .Consensus was not reached on which formulae to use when fitting hearing aids for patients with and without tinnitus , reducing stress caused by hearing difficulties, and providing distraction from tinnitus (see Table Seventy-nine percent of panelists agreed that hearing aid(s) reduce tinnitus for patients with mild hearing loss, even if there is no benefit for hearing. However, 75% also believed that hearing aids for low-frequency hearing loss are effective for tinnitus only if the tinnitus is also low frequency and vice versa. All panel members agreed that open-fit hearing aid avoids occlusion, making use of natural hearing, and 75% of members agreed that unilateral aiding can shift tinnitus perception to the unaided ear.Consensus was not reached on whether patients with mild hearing loss are more likely to stop using hearing aid(s) than patients with more significant hearing losses (<50% agreement) and whether patients with a mild hearing loss and bothersome tinnitus are more likely to use their hearing aid(s) compared with patients with a mild hearing losses but without tinnitus (64%). Sixty-one percent of clinicians disagreed with the statement that there was a trade-off between what benefit hearing aid(s) can provide for hearing loss and for tinnitus if patients have both complaints. There was no consensus as to whether questionnaires , which indicates some strongly held opinions among hearing professionals. The relatively low number of statements (58 of 115) for which consensus (\u226570% agreement) was reached and only moderate agreement between experts, as shown by Kendall\u2019s coefficient of concordance W, reflects the lack of a link between outcomes of the diagnostic assessment and choice of treatment in current practice guidelines . CliniciThe lack of high quality research evidence demonstrating effectiveness of certain tinnitus management strategies, as well as lack of linkage between assessment and diagnosis and recommended management strategies, leads to marked differences in clinical practice across different countries. In the United Kingdom and some other regions such as Scandinavia, the Netherlands, and the United States, audiology is an independent profession and the main provider of tinnitus services. However, in other countries such as Germany and Poland, audiology and therefore tinnitus services are a subspecialty of otolaryngology . As a reDepending on the country and treatment pathway, hearing aids may have a different place in the more complex management strategy. Some recent examples in the literature include progressive audiologic tinnitus management PATM,  in the UThe most common audiologic management strategy in the United Kingdom is education and reassurance, combined with sound therapy with hearing aids being the most popular choice for patients with tinnitus and a comorbid hearing loss. Where needed, it is supplemented with an intervention to reduce distress such as relaxation therapy or CBT . This waNone of the practice documents referred to earlier specify any objective audiometric criteria for fitting hearing aids for patients with tinnitus. They typically recommend only that hearing difficulties should be addressed, ignoring issues such as tinnitus pitch that may be of importance . The terOne important step in tinnitus management concerns determining whether the patient needs audiologic intervention for hearing, for tinnitus, or for both  also postulates that care must be patient centered, including creating patient-specific fitting goals. Some of the potential nonauditory predictors of success with amplification listed in the above document are the same as those that our panelists suggested might be important for the decision to fit hearing aids including motivation, realistic patient expectations, manual dexterity, cognition, prior experience with amplification, and visual acuity. However, not all of them achieved our predefined level of consensus.The Department of Health and TRI documents recommend that a diagnostic assessment of a patient with tinnitus should include a case history, audiologic assessment , psychoacoustic tinnitus measures , and assessment of tinnitus severity and psychiatric comorbidity  using standardized questionnaires . While tBoth the Department of Health and TRI also recommend the use of a tinnitus questionnaire to measure tinnitus severity. It seems that despite guidelines regarding diagnostic assessments for patients with tinnitus being relatively consistent among documents , most clStandard fitting formulae such as NAL-NL1 and more recently NAL-NL2 are the preferred formulae in NHS audiology departments (British Society of Audiology and British Academy of Audiology Guidance 2007). However, there was no consensus on which of the two is better for fitting hearing aids for a mild hearing loss and if one has an advantage over the other when the patient also has bothersome tinnitus. One reason might be that NAL-NL2 is a relatively new formula . CommentRecent work by Consensus among panelists in our study that NAL-NL1 puts emphasis on speech intelligibility and high-frequency gain needed for speech clarity is in line with the evidence provided by the above studies, as was consensus to offer separate programs for tinnitus patients: (1) with settings to optimize speech comprehension and (2) with settings to increase the audibility of background sounds. However, despite the above evidence, panelists would use the DSL formula mainly for children or patients who had previous experience with DSL. There was no consensus on whether the upset caused by tinnitus or hearing difficulty was more important to address when fitting hearing aid(s). Comments indicated that our panelists try to address both issues, but they are aware of a possible trade-off between fitting decisions that are optimized for tinnitus benefit and those optimized for listening satisfaction.Another interesting question is whether a patient with tinnitus should be offered unilateral or bilateral hearing aids. Although panelists agreed that unilateral fitting is not appropriate for patients with tinnitus, this conviction is not well evidence based.Some authors report an improvement in tinnitus handicap with both unilateral and bilateral hearing aids regardless of the laterality of tinnitus . Other aHere, the panelists indicated that for patients who do not have a bothersome tinnitus they would base their decision to offer unilateral or bilateral aid(s) on patient preference. However, in the case of a bothersome tinnitus, the consensus was that unilateral fitting of a hearing aid in patients with a bothersome tinnitus is not appropriate even if the patient has a unilateral tinnitus or an asymmetric hearing loss. Comments indicated that clinical observations suggest that unilateral fitting of hearing aids in patients with tinnitus may \u201cshift\u201d the tinnitus sensation to the unaided ear consistent with the observation by Differences in practice in the management of mild hearing loss with and without tinnitus are also seen in the choice of assessments that clinicians considered important to perform to verify hearing aid(s) fitting. While consensus was reached for most measures, there was no agreement on the safety of performing REMs at the highest loudness level (80 dB SPL), testing ULLs, and loud sound check for patients with tinnitus. Some panelists commented that performing these assessments could exacerbate tinnitus. The BSA-recommended procedure for the determination of ULLs states that they are generally helpful for hearing aid fitting, although most hearing aid fitting software contains normative ULL values that may be used instead . At the The presence of tinnitus did not influence the choice of technology/hearing aid features for a mild hearing loss. In line with the Department of Health guidelines (2007), open ear tip technology that avoids occlusion and makes use of residual hearing would be used for a mild hearing loss with and without tinnitus .This study reports on the consensus for hearing aid candidature and fitting in cases of mild hearing loss with and without bothersome tinnitus. Hearing professionals generally support the use of such technologies especially when the patient reports hearing difficulties and has realistic expectations of the technology and a bothersome tinnitus. Practices such as the use of open-fit technology and bilateral fitting of hearing aids for patients with bothersome tinnitus would seem to constitute \u201cusual care.\u201d Common limitations of guideline documents for diagnosis and management of tinnitus are the lack of a clear link between the outcomes of the diagnostic test battery and the recommended treatment, as well as guidelines being too general and lacking reference to the appropriate assessment tools , which leads to variability in clinical practice.We thank members of the Delphi expert panel for participating in the study and clinicians form Nottingham Audiology Services for their help in developing questionnaires used in this study. We also thank Vicky Owen for providing statistical support."}
+{"text": "Bacillus anthracis Tyrol 4675, isolated from a diseased cow.In 1988, an outbreak of anthrax occurred among cattle in the Austrian state of Tyrol. Since then, Austria has been declared anthrax-free. Here, we report the draft genome sequence of one of these last outbreak strains,  Bacillus anthracis strain Tyrol 4675 was isolated from one of the diseased cows  using Nextera version 3 with 2 \u00d7 300-bp chemistry. For the WGS library, 2 \u00d7 813,272 reads accumulating to a total of 464 Mb were generated. Burrows-Wheeler Aligner\u2019s Smith-Waterman Alignment (BWA-SW) , respectively. The G+C content was calculated using an in-house python script.Whole-genome shotgun (WGS) sequencing of (BWA-SW)  was usedB.\u00a0anthracis Tyrol 4675 was 5,227,565\u00a0bp, with 62.3-fold coverage for the chromosome, 145-fold for pXO1, and 102-fold for pXO2, respectively. The mean G+C content was 35.4%. For initial annotation, assembled contigs were submitted to the RAST Annotation Pipeline RASTk . Within the genome annotation, there are 11 copies of genes for the 16S rRNA, the 5S rRNAs, and the 23S rRNA; 95 tRNA loci were identified.The total length of the genome draft sequence of CP018904 (pXO1), CP018905 (pXO2), and CP018903 (chromosome). The versions described in this paper are the first versions.This whole-genome shotgun project has been deposited at DDBJ/EMBL/GenBank under the GenBank accession numbers"}
+{"text": "Organic-inorganic lead halide based perovskite solar cells have received broad interest due to their merits of low fabrication cost, a low temperature solution process, and high energy conversion efficiencies. Rare-earth (RE) ion doped nanomaterials can be used in perovskite solar cells to expand the range of absorption spectra and improve the stability due to its upconversion and downconversion effect. This article reviews recent progress in using RE-ion-doped nanomaterials in mesoporous electrodes, perovskite active layers, and as an external function layer of perovskite solar cells. Finally, we discuss the challenges facing the effective use of RE-ion-doped nanomaterials in perovskite solar cells and present some prospects for future research. This change boosted efficiency to 10.9%. It also demonstrated that the perovskite not only plays a role in sensitizers, but also can transport the electrons between different cell proportions [2 heterojunction solar cell. They directly evaporated a gold film on the top of perovskite as the electrode and reported efficiency of 7.3%. This result indicated that the perovskite can also assume the role of a hole conductor [With the constant development of human civilization, energy shortage and environmental pollution have become more and more significant challenges in our world, and they are issues that need to be resolved urgently. Solar energy is an ideal alternative energy source as it is renewable, inexhaustible, and clean energy. Gr\u00e4tzel indicated that the supply of energy from the sun to the earth is gigantic, or about 10,000 times more than the global population currently consumes , perovskportions . In the onductor .Perovskite solar cells have been regarded as one of the most ideal alternatives to silicon solar cells because of their advantageous features such as their large light absorption coefficient, high charge carrier mobility, and high conversion efficiency. However, there are some critical challenges for improving photovoltaic performance and stability of PSCs especially regarding conversion efficiency. The perovskite sensitizer is limited by its bandgap (1.55 eV), which results in its absorption spectrum rising to 780 nm . On the 3+-Er3+ co-doped glasses for lasers, Auzel found a visible green emission arising from IR excitation [n configuration, known as the f\u2013f transition. The other is the transfer of a 4f electron into the 5d sub-shell. The photoluminescence caused by an f\u2013f transition has the characteristics of narrow emission peaks, small temperature quenching, some affect by the matrix, rich emission lines, and more. Meanwhile, the f\u2013d transitions have sizeable intensities and their energies depend on the metal-ion surrounding environment [In 1966, while working on Ybcitation . Since tironment . Contribironment ,15,16.Besides the upconversion (UC) property of RE-ion-doped luminescent nanomaterials, which generate high-energy visible light after absorbing low-energy NIR light, downconversion (DC) emission of emitting two or more low-energy photons by taking up one high-energy photon may also be observed. Because of the specific band diagram of RE ions, both of the UC and DC luminescent nanomaterials have emission spectra in the range of 400\u2013700 nm, which coincides with the absorption of PSC. Therefore, the application of rare-earth ion doped luminescent nanomaterials in PSC is beneficial for the efficiency improvement of PSC.2, SnO2, ZnO, and so on. The mesoporous TiO2 film not only serves as an electron accepting and transporting layer (ETL) in PSCs, but also serves as a scaffold layer [2O3 and ZrO2, have also been investigated for creating the mesoporous film [2O3 film only served as a scaffold layer in PSC without the function of charge transportation between perovskite and Al2O3. Perovskite is used as a sensitizer to convert photons into electrons. Besides this, electron transfer and hole transfer can also occur in the perovskite layer. The HTM should be filled with the mesoporous layer to induce a heterojunction. The HTM is served as hole extracting from perovskite and can transfer the hole into the electrode. Because the PSC was approved for working without a mesoporous oxide layer, the planar pin junction structure with n-type thin TiO2, perovskite film, and p-type HTM film was constructed , the PCE was enhanced by about 7.9% with LiYF4:Yb3+, Er3+ single crystal layer on PSC structure under the irradiation of simulated solar illumination (AM 1.5 G) with an intensity of 0.73 W\u00b7cm\u22122 (about seven or eight times that of solar power). Moreover, the researchers found that, when the intensity of simulated solar illumination is more than 0.2\u20130.73 W\u00b7cm\u22122, the utilization of LiYF4:Yb3+, Er3+ crystal in PSC is active for enhancing the efficiency of PSC.The fabrication of an UC layer on a PSC is a straightforward pathway to broaden the absorption of PSCs. In 2016, LiYFn et al. . The phole range . As show2) of PSC is the most popular way for using UC material in PSCs. The advantage of using UC material in PSCs is not only to broaden the absorption spectrum of PSC, which mainly depends on the UC process of RE-doped material, but also to improve the optical absorption of PSCs contributed by the scattering effects of the UC nanoparticles.The introduction of RE-doped upconversion material into the commonly applied mesoporous electron transport layer (ETL) (such as m-TiO4:Yb/Er upconversion nanoparticles (UCNPs) to replace the m-TiO2 as the mesoporous layer in PSCs for the first time [4:Yb/Er nanoparticles, the thickness of UCNPs mesoporous layer exceeding 150 nm led to the reduction in Jsc, Voc, and PCE. (Jsc is the short circuit current density and Voc means the open circuit voltage of the solar cells).In 2016, M. He et al. reported the utilization of monodisperse NaYFrst time . By usinrst time , which i4:Yb3+, Er3+ hexagonal nanoprisms (NYF) mixed with m-TiO2 as an upconverting mesoporous layer, which can absorb the incident NIR light and convert it into visible light that can be used in PSCs [2 mesoporous layer with 75 wt % NYF. This was a 13.47% enhancement in PCE compared with NYF free devices. The enhancement of PSC performance was attributed to (a) the larger number of carriers generated in perovskite via NIR photon upconversion of NYF, and (b) the slight improvement of charge migration in PSCs due to the electrons excited in perovskite having a high chance to be transferred through high crystalline NYF rather than through the TiO2 layer.In the same year, J. Roh et al. developed and reported on the NaYF in PSCs . The bes4:Yb3+, Tm3+/NaYF4 nanoparticles (NYF NPs) which had a high luminous intensity at visible range when incorporated into the m-TiO2 ETL in PSCs [2 ratio was 0.25, which is about 20% enhancement when compared to the NYF-free PSCs. For better understanding of the mechanism behind using NYF in PSC for efficiency enhancement, the authors analyzed the field distribution from NYF NPs in PSCs with FDTD simulation. The FDTD simulation results indicated that the UC effect of NYF NPs and the light scattering by NYF NPs are both advantageous for enhancing PCE in PSCs.In 2016, M. Que et al. proposed the use of core-shell \u03b2-NaYF in PSCs . Optimal2 shell on the outside and positioning an internal \u03b2-NaYF4:Yb3+, Er3+ (NYEY) UC core to enhance the response of PSCs [4:Yb3+, Er3+@SiO2 (NYEY@S) nanoparticles were dispersed into the m-TiO2 ETL with a specific ratio of 0.5. The recorded PCE of NYEY@S incorporated PSC was 9.92%, which had an enhancement of 21% compared to the NYEY@S free devices and about 97% compared to the NYEY incorporated PSC.The major challenge of using RE-doped upconversion material in solar cells is that the defect and ligands on the surface of RE-doped upconversion material would trap a large part of photo-generated electrons and decrease the photocurrent in solar cells. To overcome this issue, J. Hu et al. proposed a new configuration of UC material involving insulating SiO of PSCs . The \u03b2-N2O3 by additionally incorporating heavily doped semiconductor material Cux2\u2212S nanoparticles, which displayed an increase in UC efficiency (14.3% under 980 nm excitation) and also broadened the emission spectra and UC excitation spectra [x2\u2212S@SiO2@Er2O3 (mCSE) nanocomposites had a lower threshold power and a broader absorption range compared to normal UC material due to the localized surface plasmon resonance (LSPR) of mCux2\u2212S nanoparticles. The authors dispersed mCSE nanocomposites in the m-TiO2 ETL with a concentration range of 0\u201310 wt %. The recorded PCE was 17.8% at an optimal mCSE/m-TiO2 ratio of 0.5 wt %, which was 10% higher than the mCSE-free devices (16.2%). To better demonstrate the improved performance of PCE, the incident photon-to-current conversion efficiency (IPCE) spectra with mSCE demonstrated the broader generation of the photocurrent range (370~1000 nm) than the devices without mSCE (370~820 nm). As shown in 2 (about \u22124.2 eV). This indicated that the mCSE nanocomposites absorbed the NIR light, generated the photo-generated electrons, and then transferred the electrons to the conduction band of m-TiO2 to increase the photocurrent of PSCs.In 2017, D. Zhou et al. developed a lanthanide oxides Er spectra . The mCu2 by the RE ion doped procedure. In 2006, W. Wang et al. prepared Er3+-doped TiO2 with sol-gel method using tetrabutyl titanate as a precursor [3+/Yb3+ co-doped TiO2 through sol-gel method and when the concentration of Yb3+ was 2.5 mol%, the intensity of fluorescence spectroscopy 543 nm. Conversion was the strongest under 980 nm excitation [2(Er-Yb:TiO2) nanorod arrays as ETL. The PCE of PSC based on the Er-Yb:TiO2 presented an enhancement of 20.8% compared to the Er-Yb:TiO2 free devices.This shows the effective progress in advancing the photoelectric properties of TiOrecursor . Under 9citation . For PSC3NH3PbI3, was the most efficient way for improving photoelectric performance of PSCs. Unfortunately, the surfaces of most as prepared RE-doped upconversion nanomaterials usually have the long-chain alkyl molecules on the surface such as oleic acid (OA) and oleylamine (OM) [The introduction of RE-doped upconversion nanomaterial into the commonly applied active layer of PSC, such as CHine (OM) , which line (OM) .4:Yb, Er (IR806-UCNCs) was fabricated by a two-step ligand-exchange approach to replace the OAm ligands on the surface of UCNCs with BF4\u2212 and improve the stability of IR806-UCNCs in DMF , and the PCE was enhanced from 12.79 to 19.70% compared to its counterpart without UCNCs. Under 980 nm laser with an intensity of 3 W\u00b7cm\u22121, a PCE of 0.37% was recorded. Besides the ligand-exchange strategy, Laura et al. proposed the use of cucurbit [3NH3PbBr3 perovskite nanoparticles (PK) closely to NaYF4:Yb3+, Tm3+ upconversion nanoparticles (UCn), which lead to the development of UCn@PKCB nanohybrids [n to PK under NIR light excitation by analyzing the emission spectra at \u03bbexc = 975 nm of UCn and UCn@PKCB.In 2017, a hydrophilic IR806 dye-sensitized upconversion nanocrystal IR806-\u03b2-NaYF DMF see a. It wasrst time . As seenohybrids . The aut3+, Ce3+, and Lu2+) have a strong absorption at the ultraviolet range. It is beneficial for RE ions to excite at the ultraviolet range and accomplish DC luminescence. Consequently, the application of RE-doped DC nanomaterials in PSC have been proposed for optimizing the stability of solar cells by reducing thermalization losses.The downconversion  process demonstrated that RE ions can result in two or more low-energy photons for absorbing one high-energy incident photon . There aIn 2002, T. Trupke et al. proposed the use of DC layers placed in front of the silicon solar cells. The researchers presented a theoretical maximum conversion efficiency of 38.6% according to a detailed balance model for a solar cell system . A signi2:Eu3+ nanophosphors by hydrothermal method and proposed it in 2017 as a potential DC material to improve the performance and light stability of PSCs [2:Eu3+ nanophosphors into m-TiO2 ETL of PSC and an energy conversion efficiency of 10.8% was recorded, which had an increase of 35% compared to the referenced energy conversion efficiency without CeO2:Eu3+ nanophosphors. In addition, the PSC added with CeO2:Eu3+ nanocrystals revealed better stability under UV illumination compared with the CeO2:Eu3+ free devices. In the same year, Ling Jiang et al. developed and reported a transparent luminescent downconverting layer (LDL) by using Eu-4,7-diphenyl-1,10-phenanthroline (Eu-complex) to enhance the response of PSCs. The LDL was coated on the back of a FTO glass of PSCs [Jsc after UV irradiation for 10 h while the Jsc of the PSCs coated with LDL only degrading to 74% at the same condition. These measurements indicated that damage to the structure of perovskite solar cell by coating with LDL on the back could be alleviated after prolonged absorption of UV light and improved stability of PSCs.For application of RE-doped DC nanomaterials in PSC, Wenhan Chen et al. prepared the uniform and efficient CeO of PSCs . The aut of PSCs . When th2 in PSCs has not been clearly illuminated since M. He et al. [2 in PSCs are important to boost the performance of PSCs. In addition, the major mechanism (scattering or photoluminescence) regarding performance enhancement in PSCs with RE-doped nanomaterials is not clear and more research will be necessary. Meanwhile, optimizing the device for introducing the RE-doped nanomaterials into PSCs is also an efficient way to develop high-performance PSCs. Based on the improvement in study of PSCs with RE-doped nanomaterials, the efficiency of PSCs will need further research.Significant improvement in experimental observation has been reported for the application of RE-doped nanomaterials in PSCs. However, there are still many issues for applying RE-doped nanomaterials in PSCs that need to be addressed. One important challenge is the low efficiency of existing RE-doped nanomaterials. Until now, there is no significant luminescence of RE-doped nanomaterials due to UC or DC under natural solar irradiation (AM 1.5). Additionally, the energy transfer mechanism between RE-doped nanomaterials and perovskite or m-TiOe et al.  introduc"}
+{"text": "Insecticide resistance threatens effective vector control, especially for mosquitoes and malaria. To manage resistance, recommended insecticide use strategies include mixtures, sequences and rotations. New insecticides are being developed and there is an opportunity to develop use strategies that limit the evolution of further resistance in the short term. A 2013 review of modelling and empirical studies of resistance points to the advantages of mixtures. However, there is limited recent, accessible modelling work addressing the evolution of resistance under different operational strategies. There is an opportunity to improve the level of mechanistic understanding within the operational community of how insecticide resistance can be expected to evolve in response to different strategies. This paper provides a concise, accessible description of a flexible model of the evolution of insecticide resistance. The model is used to develop a mechanistic picture of the evolution of insecticide resistance and how it is likely to respond to potential insecticide use strategies. The aim is to reach an audience unlikely to read a more detailed modelling paper. The model itself, as described here, represents two independent genes coding for resistance to two insecticides. This allows the representation of the use of insecticides in isolation, sequence and mixtures.The model is used to demonstrate the evolution of resistance under different scenarios and how this fits with intuitive reasoning about selection pressure. Using an insecticide in a mixture, relative to alone, always prompts slower evolution of resistance to that insecticide. However, when resistance to both insecticides is considered, resistance thresholds may be reached later for a sequence relative to a mixture. Increasing the ability of insecticides to kill susceptible mosquitoes (effectiveness), has the most influence on favouring a mixture over a sequence because one highly effective insecticide provides more protection to another in a mixture.The model offers an accessible description of the process of insecticide resistance evolution and how it is likely to respond to insecticide use. A simple online user-interface allowing further exploration is also provided. These tools can contribute to an improved discussion about operational decisions in insecticide resistance management. Insecticide resistance is a problem for malaria control \u20133 other The WHO produced a Global Plan for Insecticide Resistance Management in malaria vectors (GPIRM) , which iA recent comprehensive review of strategies to avoid resistance evolution across pesticides and drugs  concludeThis modelling approach focuses on the change in frequency of a single resistance gene for each insecticide. It assumes that genes conferring resistance are already present in the population and that resistance to each insecticide is coded by a single gene. These assumptions are consistent with previous modelling work . In suppThe simulation represents a population of randomly mixing individuals using standard population genetic approaches to avoid the need to follow every individual. One gene or locus is represented for each insecticide. Each individual has two copies of the gene and there are two potential alleles per locus. Each allele confers either resistance or susceptibility to the insecticide. Thus individuals can either have both resistance genes (RR homozygous), both susceptible genes (SS) or one of each . The combination of the two genes is termed the genotype. There are 3 genotypes when considering one insecticide and 9 genotypes (e.g. SSRR) when considering two insecticides Fig.\u00a0. FitnessThe calculation of fitness for each genotype when considering just a single insecticide is illustrated in Fig.\u00a0\u2018Dominance of resistance\u2019 determines how the fitness for SR sits between that of SS and RR for those that are exposed to the insecticide. With a value of 0 SR has the same fitness as SS (resistance is recessive) and a value of 1 SR has the same fitness as RR (resistance is dominant). For those that are not exposed, fitness of SS is set to 1 by definition and \u2018Cost of resistance\u2019 determines the fitness of RR. \u2018Dominance of cost\u2019 determines how the fitness for SR sits between that of SS and RR in the absence of the insecticide Fig.\u00a0.The simulation proceeds through non-overlapping generations. In each generation selection is represented by multiplying genotype frequencies in the population by their relative fitness, the latter depending on their absolute fitness Fig.\u00a0 and leveSo far this description just considers a single insecticide and associated resistance allele. In the model itself a second insecticide and resistance allele are included and overall fitness is calculated by multiplying the two results. This allows the model to represent populations exposed to two insecticides together in a mixture.The model outputs the change in resistance allele frequencies over time measured in generations. The number of generations at which a resistance allele frequency of 50% is reached is recorded and termed \u2018time-to-resistance\u2019. Using a 50% threshold is consistent with previous modelling analyses e.g. , 19). Pr. Pr19]).Single insecticide.Two insecticides used in sequence, replacing the first with a second once the frequency of resistance alleles reaches a threshold of 50%.Two insecticides used in a mixture (concurrently).Three insecticide use strategies are investigated.A major aim of this paper is to use modern techniques to make the modelling accessible to a wider community. The model is implemented in R , the codThe results start with a description of model runs with a single insecticide which are more straightforward and move on to two insecticides in sequences and mixtures.For single insecticide use, higher values of insecticide effectiveness, exposure, resistance restoration and dominance of resistance all resulted in faster resistance spread Fig.\u00a0a\u2013d and tCost of resistance Fig.\u00a0b had theWhen two insecticides are used, a comparison can be made between the relative performance of concurrent use in a mixture with sequential use. Each panel in Figs.\u00a0Increasing exposure (the proportion of insects that come into contact with the insecticides) Fig.\u00a0c decreasThe result is that a more effective insecticide increases time-to-resistance when used in a mixture  as described in Table\u00a0Exposure, the proportion of mosquitoes that are exposed to the insecticide, is a property of the location and will depend on the use of nets or IRS and mosquito behaviour. The frequency of resistance alleles can be measured, although when at low levels a large number of mosquitoes would need to be sampled to detect low frequencies.Anopheles gambiae data are extracted for pyrethroid (Alpha-cypermethrin) resistance associated with the kdr mutation  is  is 25] iThere are other model behaviours that can be changed by modifying model inputs within the computer code. For example, here it is assumed that insecticide interaction in a mixture is multiplicative, e.g. if the probability of surviving exposure to insecticide A alone is 0.3 and of surviving insecticide B alone is 0.2, then the probability of surviving exposure to a mixture of A and B is 0.3\u00a0*\u00a00.2\u00a0=\u00a00.06. This is consistent with recent work on aphids , althougIn addition to these uncertainties regarding resistance it is important to remember that this model examines solely the evolution of resistance and not the control of mosquitoes or malaria. Different insecticide use strategies, as well as affecting the evolution of resistance, will also influence both the numbers of vectors and their propensity to transmit the disease . There aThis model would predict that the best strategy for limiting the development of resistance in the short term would be to use no insecticide. Of course, using no insecticide is likely to have serious consequences for mosquito and malaria abundance. The challenge is to develop good strategies for delivering mosquito and malaria control in the short term while sustaining their operational capacity in the longer term by restricting the development of resistance.This paper describes a flexible, modern model of the evolution of insecticide resistance that can help us understand the mechanisms by which different insecticide application strategies may be favoured. It replicates previous modelling work and has the potential to investigate further operational strategies for established and new insecticides. For assessing the relative benefits of mixtures and sequences it demonstrates that mixtures are likely to be favoured by high insecticide effectiveness, low exposure and high costs of resistance. Similar processes will occur in other strategies such as rotations and mosaics. This accessible modelling approach can help to promote discussion and exploration of the likely effect on resistance evolution of different insecticide use strategies in tandem with the collection of empirical data to refine and test predictions."}
+{"text": "Tyrosinase is a multi-copper enzyme which is widely distributed in different organisms and plays an important role in the melanogenesis and enzymatic browning. Therefore, its inhibitors can be attractive in cosmetics and medicinal industries as depigmentation agents and also in food and agriculture industries as antibrowning compounds. For this purpose, many natural, semi-synthetic and synthetic inhibitors have been developed by different screening methods to date. This review has focused on the tyrosinase inhibitors discovered from all sources and biochemically characterised in the last four decades. Tyrosinase is the main enzyme recognised as responsible for this enzymatic browning and melanogenesis in mammalsin vitro detection methods for rapidly screening tyrosinase inhibitors may be achieved through using virtual screening,in vitro analysis will be useful to understand the functional mechanisms of the tested compounds,,,,51\u201362.Additionally, further improvements of The present review also focuses on the tyrosinase inhibitors discovered from all sources, including synthetic compounds, extracts and active ingredients of natural products, virtual screening and structure-based molecular docking studies published in the last four decades. We hope that the knowledge offered in this review serves as an updated comprehensive database contributing to the development of new safe and efficient anti-tyrosinase agents for the prevention of browning in plant-derived foods, seafood and hyperpigmentation treatments.,Melanins, the main pigment primarily responsible in the skin, hair and eyes pigmentation of human, are produced by melanocytes through melanogenesis. Melanogenesis and skin pigmentation are the most important photoprotective factor in response to ultraviolet radiation damaging from the sun and skin photo-carcinogenesis. The abnormal loss of melanin and depigmentation can be a serious facial esthetic and dermatological problem among humanAgaricus bisporus is a major and cheap source of tyrosinase with high similarity and homology compared to human tyrosinase,Agaricus bisporus is a 120\u2009kDa tetramer with two different subunits, heavy and lightAgaricus bisporus (H-subunit) have been purified with a high specific tyrosinase activity of more than 38,000\u2009U/mgTyrosinases have been isolated and purified from different sources such as some plants, animals and microorganisms. Although many of them (such as human) have been sequenced, only few of them have been characterised. Recently, a novel tyrosinase produced by Sahara soil actinobacteria have been isolated and biochemically charactrised with the aim to identify novel enzymes with exclusive features for biotechnological applications,l-tyrosine) to o-diphenols (e.g. l-dopa) and a diphenolase activity where tyrosinase oxidises o-diphenols to o-quinones (o-dopaquinone). At the same time of these enzymatic reactions, there are different chemical reactions coupled where two molecules of o-dopaquinone react their-selves generating an o-diphenol molecule (L-dopa) and a dopachrome molecule.Tyrosinase (EC 1.14.18.1) has two activities in its catalytic cycle, see Figure 2o-diphenol (see mE) binds the o-diphenol (D) originating the complex mED. This complex oxidises the o-diphenols transforming it to o-quinone and the enzyme is converted into the form deoxy-tyrosinase (dE). dE has a very big affinity for the molecular oxygen originating the form oxy-tyrosinase (oxE), which binds another o-diphenol molecule and originating the complex oxED. After that, the o-diphenol is oxidised again to o-quinone and the form Em is formed again completing the catalytic cycle. However, after these enzymatic reactions, two o-quinone molecules (e.g. o-dopaquinone) react generating dopachrome and regenerating a molecule of o-diphenol.Diphenolase activity can be independently studied, when tyrosinase reacts with an enol see . The foro-diphenol in reaction medium and is proportional to the quantity of monophenol used. oxEM (oxy-tyrosinase bound to monophenol) and mEM (met-tyrosinase bound to monophenols). oxEM is active and is transformed into mED, which is an intermediate of the catalytic cycleo-Quinones formed by these two oxidation cycle spontaneously react with each other to form oligomersAs mentioned before, we can independently study the diphenolase activity. However, it is not applicable for the monophenolase activity, see l-dopa, and activity is assessed based on dopachrome formation.Due to the critical role of tyrosinase in the melanogenesis and browning process, several investigations have been reported for the identification of tyrosinase inhibitor from both natural  and synthetic sources so far. General speaking, tyrosinase inhibitors are examined in the presence of a monophenolic substrate such as tyrosine or a diphenolic substrate such as o-dopaquinone scavengers, alternative enzyme substrates, nonspecific enzyme inactivators and denaturants, only specific tyrosinase inactivators and reversible inhibitors actually bind to the enzyme as true inhibitors and really inhibit its activity:Specific tyrosinase inactivators. They are called suicide inactivators or mechanism-based inhibitors. This group of compounds can be considered very interested from a pharmacological point of view, in hyperpigmentation processes 98.FigurAmong different types of compounds such as specific tyrosinase inactivators and inhibitors, ,To explain the suicide inactivation of tyrosinase, mainly two mechanisms have been proposedo-diphenolic substratesl- and d-dopao-diamines,,The studies about the kinetics of suicide inactivation of tyrosinase have been carried out with several o-diphenols,o-diamines,d-tyrosine negatively regulates melanin synthesis by inhibiting tyrosinase activity, competitivelyl-tyrosine has been shown as an inhibitorGenerally, the mode of inhibition by \u201ctrue inhibitors\u201d is one of these four types: competitive, uncompetitive, mixed type (competitive/uncompetitive), and noncompetitive. A competitive inhibitor can bind to a free enzyme and prevents substrate binding to the enzyme active site. Regarding the property that tyrosinase is a metalloenzyme, copper chelators such as many aromatic acids, phenolic and poly-phenolic compounds, a few non-aromatic compounds, can inhibit tyrosinase competitively by mimicking the substrate of tyrosinaseThe chemical structure of the different substrates is diverse, but the process always requires a step of oxidation/reduction: p-alkylbenzaldehydesIn contrast, an uncompetitive inhibitor can bind only to the enzyme-substrate complex and a mixed (competitive and uncompetitive mixed) inhibitor can bind to both forms of free enzyme and enzyme-substrate complex. Finally, noncompetitive inhibitors bind to a free enzyme and an enzyme\u2013substrate complex with the same equilibrium constant50 value (the concentration of inhibitor at which 50% of your target is inhibited) should be calculated in the enzyme kinetics studies and inhibitor screening to compare the inhibitory strength of an inhibitor with others. However, the IC50 values may be incomparable due to the varied assay conditions  but a positive control can be used for this purpose50 and have not applied a positive control in their studies but, fortunately, in most studies conducted for screening new tyrosinase inhibitors, the popular whitening agents, such as kojic acid, arbutin or hydroquinone, were used as a positive control,l-ascorbic acid, ellagic acid and tranexamic acid have been reported as skin-whitening agents in the cosmetic industry but there are a few reports failed to confirm their effect as an agent to lighten skin in clinical trials despite the safety of this compound50\u2009>\u2009500\u2009\u00b5mol/L) weekly inhibits human tyrosinase. In contrast, a resorcinyl-thiazole derivative, thiamidol, is a most potent inhibitor of human tyrosinase (IC50 of 1.1\u2009\u00b5mol/L) but inhibits mushroom tyrosinase weakly (IC50\u2009=\u2009108\u2009\u00b5mol/L)in vivo skin lightening potency, have been reported due to its increased skin penetration and binding affinity to human tyrosinaseIn addition to determining the inhibition mechanism, inhibitory strength which is expressed as the ICNatural sources including plants, bacteria and fungi have recently become of increasing interest for their antityrosinase activity by producing bioactive compounds. A number of researchers prefer to identify inhibitors from natural sources due to their less toxicity and better bioavalibility, especially for food, cosmetic and medicinal applications.Asphodelus microcarpusMorus nigra LGreyia radlkoferi SzyszylLimonium tetragonumArctostaphylos uva-ursiPleurotus ferulaeAgastache rugosa Kuntze fermented with Lactobacillus rhamnosus and Lactobacillus paracaseiArtemisia aucheri BoissCassia toraS. brevibracteata subspRhodiola crenulata, Alpinia officinarum Hance and Zanthoxylum bungeanum MaximMangifera indicaPodocarpus falcatusMomordica charantiaCymbopogon citratesGreyia flanaganii (IC50\u2009=\u200932.62\u2009\u00b5g/ml)Vitis vinifera Leaf extracts (IC50\u2009=\u20093.84\u2009mg/mL)Inula britannica L.,Achyrocline satureioides, Artemisia verlotiorum, Cotoneaster glaucophylla, Dalea elegans, Flourensia campestris, Jodina rhombifolia, Kageneckia lanceolata, Lepechinia floribunda, Lepe-chinia meyenii, Lithrea molleoides, Porlieria microphylla, Pterocaulon alopecuroides, Ruprechtia apetala, Senna aphylla, Sida rhombifolia, Solanum argentinum, Tagetes minuta, and Thalictrum decipiens. Besides, plants from the Moraceae family including genera Morus species, Artocarpus, Maclura (Cudrania), Broussonetia, Milicia (Chlorophora), and Ficus have shown in vitro tyrosinase inhibitionArdisia elliptica ThunbPhyllanthus acidus (L.) Skeels, Rhinacanthus nasutus L. Kurz , Arbutus andrachne L. (IC50\u2009=\u20091\u2009mg/mL)Withania somnifera L. Dunal and Solanum nigrum L. berriesPulmonaria officinalis and Centarium umbellatumBauhinia vahlii)Hypericum laricifolium Juss, Taraxacum officinale F.H.Wigg. , and Muehlenbeckia vulcanica Meisn  showed the greatest anti-tyrosinase activityIt is well known that phenolic compounds are the largest group of phytochemicals found in plants, which are mainly the factors responsible for the activities in plant extractsAspergillus sp.Trichoderma sp.Paecilomyces sp.Phellinus linteusDaedalea dickinsiiDictyophora indusiataNeolentinus lepideusMyrothecium sp. isolated from algaePestalotiopsis sp. Z233Streptomyces sp., such as S. hiroshimensis TI-C3 isolated from soilStreptomyces swartbergensis sp. Nov.Streptomyces roseolilacinus NBRC 12815Thalassotalea sp. Pp2-459Oscillatoria agardhiiLactobacillus sp.Fungi from different genera such as Finally, in an updated review by Fernandes from reported findings, tyrosinase inhibitors produced by microorganisms have been summarisedPhenolic compounds which are characterised by having at least one aromatic ring and one (or more) hydroxyl group can be classified based on the number and arrangement of their carbon atoms. These compounds are commonly found to be conjugated to sugars and organic acids. Phenolics range from simple to large and complex tannins and derived polyphenols due to their molecular-weight and number of aromatic-rings,,,,Rhus succedanea, can inhibit the activity of tyrosinase and suppress melanin production in animal cells. The IC50 of this compound (37\u2009\u00b5M) is less than hydroquinone (70\u2009\u00b5M) as a known inhibitor of tyrosinase. They have suggested that the potent inhibitory effect of this derivative on tyrosinase activity is likely due to its heptadecenyl chain, which facilitates the oxidation of the hydroquinone ring,The simple phenols such as hydroquinoned-glucopyranoside isolated from Isotachis japonica and Protea neriifolia and its glycoside derivatives  are categorised as analogs of arbutin. However, isotachioside and arbutin could not be determined as potent inhibitor. But, glucoside, xyloside, cellobioside and maltoside derivatives, missing methyl and benzoyl groups, acted as tyrosinase inhibitors with IC50s of 417, 852, 623 and 657\u202f\u00b5M, respectively. Among these novel inhibitors, glucoside derivative (IC50\u2009=\u2009417\u202f\u00b5M) was the most potent, indicating that the structural combination of resorcinol and glucose was significant for inducing the inhibitory effectIsotachioside, a methoxy-hydroquinone-1-O-beta-,,Hydroquinone and some of its known derivatives, including \u03b1 and \u03b2-arbutin, are described as both a tyrosinase inhibitor and a substratel-tyrosine, l-\u03b1-methyl-tyrosine and tyramine are substrates of tyrosinase. o-Quinone evolves in the medium of reaction accumulating o-diphenol and this accumulation provokes that met-tyrosinase (Em) is transformed into oxy-tyrosinase (Eox), which is the active form of the tyrosinase for monophenols and o-diphenols. Therefore, tyrosinase is active with monophenols such as: umbelliferone,p-hydroxybenzyl alcoholn-butylresorcinolp-coumaric acid,mE to Eox), an o-diphenol or a reducing agent such as ascorbic acid transforming Em to Ed which, with molecular oxygen, is transformed into Eox. A particular case is deoxyarbutin, which acts as a substrate of tyrosinase even if any reagent is not added to the medium of reaction,Monophenolic compounds such as ,Plants produce a large diverse class of polyphenols including phenolic acids, flavonoids, stillbenes and lignans,,Trifolium nigrescens Subsp. PetrisaviVigna radiatae L.)calamondin peelMorus yunnanensisBhagwa and Arakta cultivarTibouchina semidecandra LMaackia fauriePleurotus ostreatusPotentilla bifurcaAlpinia officinarumMorus lhouGarcinia subellipticaArtocapus altilisMyrsine africanaPulsatilla cernuaSalvia miltiorrhiza-Carthamus tinctorius  herbal pairAmong polyphenolic compounds, some of the flavonoid derivatives mostly found in herbal plants, fruits and synthetic sources have been raveled to be the potent inhibitors of tyrosinaseLimonium tetragonum50\u2009=\u20090.87\u2009mg/ml)50\u2009=\u20097.3\u2009\u00b5M) against tyrosinase, have been approved by Lee et\u00a0al.Generally, major flavonoids  are clasArtocapus altilis with 11 other phenolic compounds for their inhibitory activities on tyrosinase50 value of 10.31\u2009\u00b1\u20090.41\u2009\u00b5M and a noncompetitive manner with a iK of 9.50\u2009\u00b1\u20090.40\u2009\u00b5M. The quenching anlaysis of tyrosinase by this compound showed a static mechanism and a single binding site with a binding constant of 7.50\u2009\u00b1\u20091.20\u2009\u00d7\u2009104 \u00a0M\u22121 at 298\u00a0K. Based on the thermodynamics parameters, the binding process involved hydrogen bonds and van der Waals forces. Also, docking simulation illustrated hydrogen bonds between this compound and the residues His244 and Met280 of active siteThe most common flavones are luteolin, apigenin, baicalein, chrysin and their glycosides 50\u2009=\u20090.11\u202fmM) indicated a mix-type . A single binding site with a binding constant of 2.78\u2009\u00d7\u2009105\u202fM \u2212 1 was obtained from the quenching fluorescence analysis for this compound. Thermodynamic parameters suggested spontaneous binding through hydrogen bonding and van der Waals forces. Furthermore, circular dichroism spectra indicated a reduction in the content of \u03b1-helix from 32.67% to 29.00% due to this binding. Docking simulations also indicated that baicalein mainly bound tyrosinase via its Met280 residueIn addition, several hydroxyflavones including baicalein, 6-hydroxyapigenin, 6-hydroxygalangin and 6-hydroxy-kaempferolMorus alba L. (IC50\u2009=\u2009290.00\u2009\u00b1\u20097.90\u2009\u00b5M)Trifolium nigrescens Subsp. Petrisavi50\u2009=\u20096.3\u2009mg/ml) and a C-glycosylflavone isovitexin (IC50\u2009=\u20095.6\u2009mg/ml) from Vigna radiatae L. extracts exhibited significant tyrosinase inhibition activities50\u2009=\u20090.088\u2009mM), cyclomorusin (IC50\u2009=\u20090.092\u2009mM), morusin (IC50\u2009=\u20090.250\u2009mM), kuwanon C (IC50\u2009=\u20090.135\u2009mM) and norartocarpetin (IC50\u2009=\u20091.2\u2009\u00b5M) isolated from the stem barks of Morus lhou (S.) Koidz, have been investigated by Ryu et\u00a0al. The mechanism of inhibition indicated that mormin, cyclomorusin, kuwanon C and norartocarpetin inhibited tyrosinase competitivelyAnother flavone named morusone from the twigs of O-glycosidesHypericum laricifolium JussCrocus sativus L.Olea europaea L.d-glucoside from Potentilla bifurcad-glucopyranoside and kaempferol-3-O--\u03b2-d-glucopyranoside from mulberry leavesAlpinia officinarum50\u2009=\u200931.1\u2009\u00b5M), 2,3-trans-dihydromorin (IC50\u2009=\u200921.1\u2009\u00b5M) from Cudrania cochinchinensisMyricetin, kaempferol, quercetin, morin, isorhamnetin, galangin and their glycosides  are the predominant flavonols most commonly found as iK\u2009=\u20094.03\u2009\u00b1\u20090.26\u2009mM and the binding of morin to tyrosinase-induced rearrangement and conformational changes of the enzymeBased on kinetics studies, morin reversibly inhibited tyrosinase through a multi-phase kinetic process and bind to tyrosinase at a single binding site mainly by hydrogen bonds and van der Waals forces. It inhibited tyrosinase reversibly in a competitive manner with Sophora flavescens50 values less than 10\u2009\u00b5M. Finally, based on the literature review, many flavonol inhibitors are usually competitive inhibitors due to the 3-hydroxy-4-keto moiety of the flavonol structure, which chelates the copper in the active site d-glucoside with a IC50 value of 1.9\u2009\u00b5M is revealed stronger tyrosinase inhibition than their positive control, kojic acidRecently, 8-prenylkaempferol as a competitive tyrosinase inhibitor along with Kushenol A (noncompetitive) isolated from 50 value of 11.21\u2009\u00b1\u20090.8\u2009\u00b5M and 5.23\u2009\u00b1\u20090.6\u2009\u00b5M, respectively, whereas very low inhibition activity was obtained for 6,7,4'-trihydroxyisoflavone, daidzein, glycitein and genistein50 value of 9.2\u2009\u00b5M, which is six times potent than kojic acidIsoflavones such as daidzein, genistein, glycitein, formononetin, and their glycosides  mostly are detected in the medicinal herbs50\u2009=\u20090.43\u2009\u00b5M), isolated from the root of Glycyrrhiza glabra Linn, has exhibited excellent inhibitory effects on tyrosinase. The quenching analysis of tyrosinase by glabridin showed a static mechanism50\u2009=\u200911.40\u2009\u00b5M) against tyrosinase. They have suggested the more effective interaction with the enzyme may be due to more conformational flexibly of this compound that has occurred by the 4-substituted resorcinol skeleton and the lacking of double bond between carbon atom 3'' and 4'' in its structureGlycyrrhiza glabra is another kind of isoflavone identified as tyrosinase inhibitorMaackia fauriei, have been investigated by Kim et\u00a0al. for their tyrosinase inhibition activity. Based on their results, among these falvonoids, mirkoin (IC50\u2009=\u20095\u2009\u00b5M) revealed stronger tyrosinase inhibition than the positive control, kojic acid and inhibited tyrosinase reversibly in a competitive mode50\u2009=\u200939.7\u2009\u00b1\u20091.5\u2009\u00b5M), and 2\u2032-hydroxygenistein-7-O-gentibioside (IC50\u2009=\u200950.0\u2009\u00b1\u20093.7\u2009\u00b5M) from Apios americana were identified as competitive inhibitors, with iK values of 10.3\u2009\u00b1\u20090.8\u2009\u00b5M and 44.2\u2009\u00b1\u20091.7\u2009\u00b5M, respectivelyRecently, a noncompetitive inhibitor, glabridin , from Morus alba LMorus nigra have been identified as potent tyrosinase inhibitors by Hu et\u00a0al.Flavanones such as naringenin, hesperetin, eriodictyol and their glycosides  and flavanonols (taxifolin) are mainly found in citrus fruits and the medicinal herbs,Alphitonia neocaledonica (Rhamnaceae) is an endemic tree of New Caledonia, which has been identified as an anti-tyrosinase source due to the presence of tannins and gallocatechinDistylium racemosum branches, with IC50 value of 30.2\u2009\u00b5g/mL, showed higher tyrosinase inhibition activity than arbutin as a positive controlClausena lansium demonstrated potent mushroom tyrosinase inhibition in a mixed competitive manner and illustrated strong inhibition of the melanogenic activity of B16 cells. The IC50 values for the monophenolase and diphenolase activities were 23.6\u2009\u00b1\u20091.2 and 7.0\u2009\u00b1\u20090.2\u2009\u00b5g/mL, respectively. Furthermore, from the inhibition mechanism of this compound, it can be concluded that a chelation between the hydroxyl group on the B ring of the proanthocyanidins and dicopper ions of the enzyme has been occurredFlavan-3-ols are the most complex subclass of flavonoids ranging from the simple monomers (+)-catechin and its isomer (\u2212)-epicatechin to the oligomeric and polymeric proanthocyanidins, which are also known as condensed tannins. Flavanols, such as catechin, epicatechin, epi-gallocatechin, epicatechin gallate (ECG), epigallocatechin gallate (EGCG) and proanthocyanidins are widespread in the medicinal herbs and higher plantsAnnona squamosa) pericarp could powerfully inhibit the activities of monophenolase and diphenolase of tyrosinase, competitivelyl-tyrosine hydroxylation and L-DOPA oxidation by chelation to the active site of tyrosinaseLongan Bark indicated the reversible and mixed (competitive is dominant) inhibition of tyrosinaseAnother investigation revealed that procyanidin-type proanthocyanidins, purified from cherimoya  and their glycosides, are widely distributed in the medicinal herbsArtocapus altilis and showed more potent tyrosinase inhibitory activities than the positive control kojic acid,m-diphenols and o-diphenols have been investigated as potent inhibitors of mushroom tyrosinaseTwo phenolic compounds, namely curcumin and desmethoxycurcumin have been isolated from the methanolic extract of the heartwood of ,In search of tyrosinase inhibitors, the inhibitory effects of several coumarin derivatives 277\u2013279 strans-N-coumaroyltyramine (IC50\u2009=\u200940.6\u202f\u00b5M) and cis-N-coumaroyltyramine (IC50\u2009=\u200936.4\u202f\u00b5M) from Humulus japonicus showed potent tyrosinase inhibitionInterestingly, among hydroxycoumarins, the 3-hydroxycoumarinMorus australis (19\u201322)Morus nigraCarex vulpinoidea seedsFlemingia philippinensis50\u2009=\u20090.07\u2009\u00b1\u20090.02\u2009\u00b5M) and morachalcone A (IC50\u2009=\u20090.08\u2009\u00b1\u20090.02\u2009\u00b5M) from Morus alba L.Psoralea corylifoliaChalcones , or 1,3-diphenyl-2-propen-1-ones, are one of the most important classes of flavonoids. Chalcone-containing plants have been used for a long time in traditional medicineustralis 19\u201322291,,Also, tyrosinase inhibitory effects of several synthetic chalcones and their derivatives were evaluated by various researchers. Oxindole-based chalconesMorus notabilisMorus yunnanensisMorus spMorus macroura (IC50\u2009=\u20090.39\u2009\u00b5M) are potent tyrosinase inhibitors among auronesOkombi et\u00a0al. have identified Z-benzylidenebenzofuran-3(2H)-one and analogs as human tyrosinase inhibitors. However, they found that aurones are weak inhibitors, but their derivatives with two or three hydroxyl groups preferably at 4,6 and 4' positions make them significant tyrosinase inhibitors. For example, the most potent aurone, 4,6,4'-trihydroxyaurone induces 75% inhibition at 0.1\u2009mM concentration and is highly effective compared to kojic acidp-coumaric, caffeic and ferulic acids. So far, p-hydroxybenzoic acid, chlorogenic acid (the ester of caffeic acid), vanilic acid (4-hydroxy-3-methoxybenzoic acid) and protocatechuic acid (a dihydroxybenzoic acid) from Hypericum laricifolium Juss50\u2009=\u20090.40 \u00b5g/mL) from Phellinus linteusp-coumaric acid from ginseng leavesm-coumaric acidp-coumarateBreynia officinalisn-nonyl esterSpiranthes sinensisKIS\u2009=\u20092.135\u2009mM and iK\u2009=\u20090.661\u2009mM309. Furthermore, n-butyl, iso-propyl, sec-butyl, n-pentyl, n-hexyl and n-octyl orsellinates  behaved as inhibitors at 0.50\u2009mM, whereas methyl, ethyl, n-propyl, tert-butyl, and n-cetyl orsellinates acted as tyrosinase activators. Thus, tyrosinase inhibition increased with chain elongation, suggesting that the enzyme site can accept an eight-carbon alkyl chainPhenolic acids are divided into hydroxybenzoates and hydoxycinnamates. The most common hydroxycinnamates are 50\u2009=\u20096.97\u2009\u00b5M) and diphenolase  activity of mushroom tyrosinase. Also, Oyama et\u00a0al. have found that some modification such as esterification can abrogate this inhibitory activity of tyrosinaseIn addition to these compounds, 3-phenylbenzoic acid (3-PBA) was revealed to be the most potent inhibitor against monophenolase Cudrania cochinchinensis (IC50\u2009=\u20092.33\u2009\u00b5M)azo-oxyresveratroltrans-resveratrol from Streptomyces avermitilis MA4680 (Gnetum gnemon),Veratrum patuluml-cysteine, which indicated suicide inhibition mechanism of resveratrolResveratrol is the most common stilbene. Several stillbenes derivatives from natural and synthetic sources  have beeCastanea henryiMarrubium velutinum and Marrubium cylleneumPinellia ternateCrataegus pinnatifidaLignans are complex and diverse structures, which are formed from three primary precursors. So far, lignans and lignan glycosides isolated from exocarp of Psoralea corylifoliaWulfenia carinthiaca JacqScabiosa stellatai value 0.05\u2009\u00b5M and IC50\u2009=\u20090.0167\u2009\u00b5MCarvacrol is a monoterpenoid phenol. To date, some carvacrol derivativesAloe veraPolygonum cuspidatum50\u2009=\u20091214\u2009\u00b5M) have been verified as tyrosinase inhibitorsThe quinones are a class of small molecules that are mostly derived from aromatic compounds such as benzene or naphthalene. Among these compounds, Aloin, an anthraquinone-C-glycoside from d-oligoxylosidesSeveral biphenyl derivativesl-phenylalanine conjugates named ((S)-(5-(benzyloxy)-1-octyl-4-oxo-1,4-dihydropyridin-2-yl) methyl 2-amino-3-phenylpropanoate) showed potent inhibitory effect with IC50 values of 12.6 and 4.0\u2009\u00b5M for monophenolase and diphenolase activities, respectivelySome hydroxypyridinone derivatives,,p-hydroxy and p-methoxy benzaldehyde thiosemicarbazonep-methoxybenzaldehyde thiosemicarbazone and 4-dimethylaminobenzaldehyde-thiosemicarbazone and 4-dimethylaminobenzaldehyde-N-phenyl-thiosemicarbazoneSeveral kinds of thiosemicarbazone derivatives50 0.42, 0.35, 0.36 and 0.44\u2009mM, respectively) than Thio-5, Thio-6, Thio-7 and Thio-8Based on the findings, the appropriate functionalisation of thiosemicarbazone may be improved the inhibitory activity of these inhibitors. Dong et\u00a0al. believe that the sterically bulky group at the C-4 position of the thiophene ring contributes to this activity. For example, the 4-functionalisation thiophene-2-carbaldehyde thiosemicarbazone with a methoxyacetyl groupd-glycosidesn-alkylp-phenylene-bis, phenylp-xylidine-bis and p-pyridine dithiocarbamate sodium salts3In addition to thiosemicarbazone derivatives, thiosemicarbazide and its derivativesSo far, several azole derivatives  have bee,Kojic acid is a well-known tyrosinase inhibitor. When DL-DOPA, norepinephrine and dopamine are oxidised by tyrosinase, Kojic acid inhibits effectively the rate of formation of pigmented product(s) and of oxygen uptake50 value of 1.35\u2009\u00b1\u20092.15\u2009\u00b5MRecently, Xie et\u00a0al. have reported a kojic acid analog namely 5-phenyl-3-[5-hydroxy-4-pyrone-2-yl-methylmercapto]-4--1,2,4-triazol as a potent competitive tyrosinase inhibitor with an ICBenzaldehydeO-ethyloxime (IC50\u2009=\u20090.3\u2009\u00b1\u20090.1\u2009\u00b5M) is of the same magnitude as one of the best tyrosinase known inhibitors tropolone (IC50\u2009=\u20090.13\u2009\u00b1\u20090.08\u2009\u00b5M)Among these derivatives, 3,4-dihydroxybenzaldehyde-Inhibitory effects of pyruvic acid, acrylic acid, propanoic acid, 2-oxo-butanoic acid, and 2-oxo-octanoic acidn-alkyl acids . While, the monophenolase activity is inhibited by all types of n-alkyl acids. They have believed that there is a physical difference in the docking of mono- and o-diphenols to the tyrosinase active site. On the other hand, the binding of acids occurs through their carboxylate group with one copper ion of the binuclear site. So these carboxylic acid compounds completely block the monophenolase reaction, by preventing monophenol binding to the oxyform of the enzymeBased on the findings investigated by Gheibi et\u00a0al., aliphatic carboxylic acids have dual effects on the monophenolase and diphenolase activities of mushroom tyrosinase. They have found that optimal diphenolase activity of tyrosinase takes place in the presence of 3H7OCS2Na, C4H9OCS2Na, C5H11OCS2Na, C2H5OCS2Na, and C6H13OCS2Na have been examined for inhibition of both monophenolase and diphenolase activities of mushroom tyrosinase. Based on the reports, C3H7OCS2Na and C4H9OCS2Na showed a mixed inhibition pattern on monophenolase activity but C5H11OCS2Na and C6H13OCS2Na showed a competitive and C2H5OCS2Na showed uncompetitive inhibition pattern. For diphenolase activity, C3H7OCS2Na and C2H5OCS2Na showed mixed inhibition but C4H9OCS2Na and C5H11OCS2Na and C6H13OCS2Na showed competitive inhibitioniK values for monophenolase inhibition and an increase of the iK values for diphenolase inhibitionThe inhibitory effect of some synthesised xanthates including C50 of 2.92\u2009\u00b1\u20090.16\u2009mM50\u2009=\u200990\u2009\u00b1\u20090.6\u2009\u00b5M50\u2009=\u20099.0\u2009\u00b1\u20091.8\u2009mM)50 of 5.57\u2009mg/mL)n-alkyl sulfates50\u2009=\u2009721.91\u2009\u00b5g\u2009mL\u22121) isolated from ribose-histidine Maillard reaction productsExcept the inhibitors listed above, other compounds have also been registered for their tyrosinase inhibitory activity by different researchers such as: two Keggin-type polyoxometalates containing glycine as potent inorganic reversible inhibitorsSynergistic strategy for tyrosinase inhibitors is a useful strategy for the improvement of their inhibitory activities. Based on the findings, the mixtures of glabridin:resveratrol, glabridin:oxyresveratrol, resveratrol:oxyresveratrol, phenylethylresorcinol:resveratrolDue to the vital role of tyrosinase in the enzymatic browning of food and depigmentation disorders in humans, its inhibitors have been considered by researchers, extensively. As mentioned above, natural sources such as plants and microorganisms and their effective compounds have wonderful potential as organic anti-tyrosinase sources.However, the majority of the compounds identified from natural sources were isolated from plants but, recently, microorganisms are considered as potential sources of tyrosinase inhibitors. It is interesting that despite the diversity of natural inhibitors, a large number of tyrosinase inhibitors are phenolic-based structures. Many researchers have designed appropriate scaffold inspired by the structure of natural compounds and developed novel synthetic inhibitors. In this paper, many natural, semi-synthetic and synthetic inhibitors have been summarised and the inhibitory effects of these compounds on the tyrosinase activity are discussed.Based on the results, phenolic compounds (simple phenols and polyphenols) and their derivatives and several compounds including terpenoid, phenyl, pyridine, piperidine, pyridinone, hydroxypyridinone, thiosemicarbazone, thiosemicarbazide, azole, thiazolidine, kojic acid, benzaldehyde and xanthate derivatives were characterised as potent tyrosinase inhibitors. The appropriate functionalisation of these inhibitors such as C-6 and C-7 hydroxyl groups of the isoflavone skeleton, 4-functionalisation thiophene-2-carbaldehyde thiosemicarbazone with a methoxyacetyl group and the aldehyde group and methoxy group in C4 of benzaldehyde derivatives may be improved the inhibitory activity of these inhibitors. Furthermore, in cholcone derivatives, the location of the hydroxyl groups on both aromatic rings and the number of hydroxyls is an important factor in the efficacy of a chalcone. In contrast, some modifications such as the prenylation or the vinylation of some flavonoid molecules do not enhance their tyrosinase inhibitory activity while deglycosylation of some flavonoid glycosides by far-infrared irradiation can be improved tyrosinase inhibitory activity. Interestingly, among different inhibitors, some compounds, especially hydroquinone and its known derivatives (\u03b1 and \u03b2-arbutin), are described as both a tyrosinase inhibitor and a substrate.in vitro and docking experiments, especially for the application of such materials in food and medicinal products. Finally, we hope that the information provided in this study, which is the result of numerous researchers\u2019 efforts, could serve as leads in the search for effective anti-tyrosinase agents from natural and synthetic sources with increased efficiency and safety in the food and cosmetics industries.Actually, the main objective of this review is to provide a useful source of effective tyrosinase inhibitors. However, despite the existence of a wide range of tyrosinase inhibitors from natural and synthetic sources, only a few of them, in addition to being effective, are known as safe compounds. Therefore, it is recommended to examine the efficacy and safety of inhibitors by in vivo models, along with"}
+{"text": "Z)-3-(3-bromo-4-hydroxybenzylidene)thiochroman-4-one (MHY1498), and evaluated its tyrosinase inhibitory activity in vitro and in silico. The chemical structures of (Z)-3-benzylidenethiochroman-4-one analogues, including the novel compound MHY1498, were rationally designed and synthesized as hybrid structures of reported potent tyrosinase inhibitors, which were confirmed both in vitro and in vivo: (Z)-5-(substituted benzylidene)thiazolidine-2,4-diones (Compound A) and 2-(substituted phenyl)benzo[d]thiazoles (Compound B). During screening, MHY1498 showed a strong dose-dependent inhibitory effect on mushroom tyrosinase. The IC50 value of MHY1498 (4.1 \u00b1 0.6 \u03bcM) was significantly lower than that of the positive control, kojic acid (22.0 \u00b1 4.7 \u03bcM). In silico molecular multi-docking simulation and inhibition mechanism studies indicated that MHY1498 interacts competitively with the tyrosinase enzyme, with greater affinity for the active site of tyrosinase than the positive control. Furthermore, in B16F10 melanoma cells treated with \u03b1-melanocyte-stimulating hormone, MHY1498 suppressed both melanin production and tyrosinase activity. In conclusion, our data demonstrate that MHY1498, a synthesized novel compound, effectively inhibits tyrosinase activity and has potential for treating hyperpigmentation and related disorders.Tyrosinase is a key enzyme in melanin synthesis, catalyzing the initial rate-limiting steps of melanin synthesis. Abnormal and excessive melanin synthesis is the primary cause of serious skin disorders including melasma, senile lentigo, freckles, and age spots. In attempts to find potent and safe tyrosinase inhibitors, we designed and synthesized a novel compound, ( Melanogenesis is the process that leads to the production of the dark macromolecular pigment melanin by melanocytes. Melanin synthesis occurs via a serial process of enzymatic catalyses and chemical reactions . The mell-tyrosine in melanin biosynthesis thiazoles (compound B) as potential tyrosinase inhibitors and demonstrated their potent inhibitory activity both in vitro and in vivo [d]thiazolyl moiety. Present in compound A, N-phenylthiourea was reported to inhibit a catechol oxidase enzyme belonging to the tyrosinase type-3 copper protein group via its sulfur atom binding to both copper ions located in the tyrosinase active site [d]thiazole compound, in which the sulfur atom of the benzothiazole ring can act as a chelator of copper ions in the active site of tyrosinase, similar to N-phenylthiourea. Additionally, because the nitrogen atom of the substituted phenyl-benzo[d]thiazole derivatives can form a quaternary ammonium ion at physiological pH, similar to tyrosine and l-DOPA, the nitrogen atom might act as a positive center capable of interacting with anionic or partially anionic groups of amino acid residues present in the tyrosinase active site. Therefore, hydroxyl- and alkoxy-substituted phenyl-benzo[d]thiazole derivatives might serve as potent competitive tyrosinase inhibitors. The imidazole ring in compound B was reported to possess significant inhibitory activity against tyrosine thiazoles (compound B) as potential tyrosinase inhibitors (in vitro and in vivo studies demonstrated that these compounds had potent tyrosinase inhibitory effects [N-phenylthiourea moiety has been reported to inhibit a catechol oxidase enzyme belonging to the group of tyrosinases that contain a copper group in the tyrosinase active site. Compound B has a 2-(hydroxyl-substituted phenyl)benzo[d]thiazolyl moiety, and its imidazole ring has been reported to exhibit notable inhibitory activity against tyrosinase. Based on the rationale that these characteristic structures may promote strong tyrosinase inhibition, hybrid structures of these two (Z)-3-(hydroxyl-substituted benzylidene)thiochroman-4-one analogs were rationally designed and synthesized to identify potent tyrosinase inhibitors -3-(hydroxyl-substituted benzylidene)thiochroman-4-one analogue compounds , were synthesized via a Claisen\u2013Schmidt condensation reaction. Appropriate hydroxyl-substituted benzaldehydes were treated with thiochroman-4-one under acidic conditions (1.0 M HCl solution in acetic acid) with or without the addition of 12.0 M HCl aqueous solution to generate the desired compounds (Z)-configuration were predominantly generated, likely because of steric hindrance. For the synthesis of the novel compound g (Z)-3-(3-bromo-4-hydroxybenzylidene)thiochroman-4-one), a solution of 3-bromo-4-hydroxybenzaldehyde and thiochroman-4-one in 1.0 M HCl in acetic acid (0.5 mL) and 12.0 M hydrochloric acid (0.02 mL) was heated at 60 \u00b0C for three days. The reaction mixture was partitioned between dichloromethane and water, and the organic layer was dried over anhydrous MgSO4, filtered, and evaporated in vacuo. The resultant residue was recrystallized in methanol and water to give MHY1498 as a yellow solid. The chemical structure of the novel compound MHY1498 is shown in As shown in ompounds , were syompounds , with yimpound g , or MHY1Z)-3-(hydroxyl-substituted benzylidene)thiochroman-4-one analogues on mushroom tyrosinase was evaluated using kojic acid as a positive control compound. As tyrosinase is an enzyme that mediates the biochemical conversion of both l-tyrosine and l-DOPA to dopachrome in melanogenesis, tyrosinase activity was evaluated by measuring the absorbance of the generated dopachrome. The inhibitory activity of the synthesized analogue compounds on mushroom tyrosinase was screened in a cell-free system. The results indicated that MHY1498 (compound g) had stronger inhibitory activity against tyrosinase than kojic acid (l-DOPA as the tyrosinase substrate (data not shown). Compared to kojic acid (10 \u03bcM), MHY1498 showed significantly decreased tyrosinase activity (data not shown). The IC50 values for MHY1498 and kojic acid were calculated as shown in 50 was 4.1 \u00b1 0.6 \u03bcM for MHY1498, and 22.0 \u00b1 4.7 \u03bcM for kojic acid, indicating that MHY1498 is approximately a five-fold more potent tyrosinase inhibitor than kojic acid. To investigate the inhibitory mechanism of MHY1498, Lineweaver\u2013Burk double-reciprocal plots were evaluated using different concentrations of l-tyrosine substrate . The change in absorbance was measured in a time-dependent manner, and the results were plotted double-reciprocally as 1/V versus 1/[S]  are shown in As tyrosinase is an essential enzyme for melanogenesis, we used in vitro data demonstrating a greater percent inhibition of MHY1498 compared to kojic acid. Although the MHY1498 inhibitory mechanism has not been clearly elucidated by X-crystallography, computational docking simulation has been established as a powerful tool to screen and evaluate new pharmacological agents. Docking simulation results provide not only the biding sites, but also the binding energies generated between a chemical compound and an enzyme, which is informative prior to the initiation of in vitro cell-based experiments. Although the predictions are not always accurate and require biological experimental verification, this method reduces time and effort when screening for effective compounds. Consequently, our data, providing predictive docking energy and revealing binding residues, support the conclusion that MHY1498 exhibits stronger tyrosinase inhibition than kojic acid because of its higher affinity for the tyrosinase active site.The multi-docking scores were generated using three different simulation programs, i.e., Autodock Vina, Autodock 4, and Dock 6. The binding energies predicted by these three programs between MHY1498 and tyrosinase were \u22126.6, \u22127.0, and \u221230.5 kcal/mol, respectively, and \u22125.6, \u22125.2, \u221227.6 kcal/mol, respectively, for kojic acid . The resThe depigmentation activity of MHY1498 was evaluated using B16F10 murine melanoma cells. The cytotoxic effect of MHY1498 was measured at various concentration of MHY1498  for 24 and 48 h a,b. The in vitro assessment was selected near the IC50 value (4.1 \u00b1 0.6 \u03bcM) for MHY1498, below the dose that did not seem to elicit cytotoxicity (10 \u03bcM). Treatment with 1 \u03bcM \u03b1-MSH induced an increase in melanin content in B16F10 melanoma cells, and co-treatment with MHY1498 marginally decreased melanin content in a dose-dependent manner , in the presence or absence of 1 \u03bcM \u03b1-MSH. The concentration for t manner a. Total t manner b. The ret manner c and wast manner d. The rel-tyrosinase, l-DOPA, \u03b1-melanocyte-stimulating hormone (\u03b1-MSH), and other chemical reagents were purchased from Sigma .Kojic acid, mushroom tyrosinase, Z)-3-(3-bromo-4-hydroxybenzylidene)thiochroman-4-one), a solution of 3-bromo-4-hydroxybenzaldehyde  and thiochroman-4-one  in 1.0 M HCl acetic acid (0.5 mL) and 12.0 M hydrochloric acid (0.02 mL) was heated at 60 \u00b0C for three days. The reaction mixture was divided between dichloromethane and water, and the organic layer was dried over anhydrous MgSO4, filtered, and evaporated in vacuo. The resultant residue was recrystallized in methanol and water to give MHY1498  as a yellow solid.For the synthesis of compound g , Autodock 4 , and Dock 6  using the crystal structure of Agaricus bisporus (PDB ID: 2Y9X) to simulate the 3D structure of tyrosinase. Docking simulations were performed between tyrosinase and either MHY1498 or kojic acid. The compounds were prepared for docking simulation, and charges were calculated. The prediction of possible hydrogen binding residues between tyrosinase and the compounds was generated using pharmacophores.For the l-tyrosine or l-DOPA was used as a substrate. As tyrosinase is an enzyme that mediates the conversion of both l-tyrosine and l-DOPA to dopachrome, enzymatic activity was monitored by dopachrome formation at 492 nm in the presence of a potent tyrosinase inhibitor. The methods used were as follows: An aqueous solution of mushroom tyrosinase  was added to the wells of transparent 96-well plates containing 200 \u00b5L of a reaction mixture of 1 mM l-tyrosine, 50 mM phosphate buffer (pH 6.5), and the test material (kojic acid or MHY1498) at different concentrations [10\u201350 \u00b5M]). The mixture was incubated at 37 \u00b0C for 30 min, and the dopachrome was determined by measuring the absorbance at 492 nm, using a microplate reader . The inhibitory mechanism of MHY1498 was evaluated using a kinetic assay with different concentrations of l-tyrosine and MHY1498.The inhibitory effect of MHY1498 was examined against mushroom tyrosinase. In the tyrosinase inhibition assay, 50 was calculated from experiments performed with different MHY1498 doses. A tyrosinase kinetic assay was performed to determine the inhibitory mechanism of MHY1498. Various concentrations of l-DOPA  were used for the inhibition assay. After examination, each value was converted into its reciprocal form following Lineweaver\u2013Burk plots. The results showed the plot of 1/V versus 1/[S], and the intersection of each plot was used to determine the inhibitory mechanism.The IC2 at 37 \u00b0C. Cell viability was analyzed using the MTT -2,5-Diphenyltetrazolium Bromide) assay. The cells were plated in each well of transparent 96-well plates at a density of 5 \u00d7 104 cells/mL and treated with MHY1498 at different concentrations, with the addition of the MTT solution. After a 1 h of incubation, the absorbance was measured at 560 nm using a microplate reader.Murine melanoma B16F10 cells were purchased from the American Type Culture Collection . The cells were cultured in Dulbecco\u2019s Modified Eagle\u2019s Medium (DMEM) containing 5% fetal bovine serum  in 5% CO4 cells/mL, incubated with MHY1498 for 2 h, and treated with 1 \u03bcM \u03b1-melanocyte-stimulating hormone (\u03b1-MSH) for 48 h. After washing twice with PBS, the cells were evaluated either for melanin content or tyrosinase activity. For melanin content, the samples were dissolved in 500 \u00b5L of 1 M NaOH, incubated at 60 \u00b0C for 1 h, and mixed to solubilize melanin. Melanin content in the samples was determined by measurement of the absorbance at 405 nm. For tyrosinase activity, the cells were lysed in 90 \u03bcL of 50 mM sodium phosphate buffer (pH 6.8) containing 5 \u00b5L of 1% Triton 100X and 5 \u03bcL of 0.1 mM phenylmethylsulfonyl fluoride (PMSF) The samples were frozen at \u221280 \u00b0C for 30 min. After thawing and mixing, cellular extracts were obtained by centrifugation at 12,000 \u00d7 g for 30 min at 4 \u00b0C. The supernatants (80 \u03bcL) with 20 \u03bcL of l-DOPA (2 mg/mL) were transferred to a 96-well plate, and absorbance was measured at 492 nm every 10 min for 1 h at 37 \u00b0C.The intracellular melanin levels were determined using B16F10 cells. B16F10 cells were plated on 35 mm dishes at 5 \u00d7 10In vitro and in silico evidence showed that tyrosinase activity was strongly suppressed, and we further recorded notable binding affinity between tyrosinase and MHY1498, suggesting that MHY1498 may be a potent tyrosinase inhibitor with potential for use as a cosmetic and therapeutic agent for hyperpigmentation-related disorders.In summary, the results of the current study indicate that the novel compound MHY1498 is a potent inhibitor of tyrosinase, a rate-limiting enzyme in melanin biosynthesis."}
+{"text": "Morus sp. are reported to exhibit anti-Alzheimer\u2019s disease, anti-inflammatory, fungicidal, anti-cancer, anti-bacterial, and anti-tyrosinase properties. We investigated the inhibition of mono- and diphenolase activity of mushroom tyrosinase by KG, MG, and AB. KG and MG displayed acceptable inhibition activity compared to kojic acid. AB did not show any activity up to 350 \u00b5M. MG displayed six-fold higher inhibition of l-tyrosine oxidation (IC50 = 6.35 \u00b1 0.45 \u00b5M) compared to kojic acid (IC50 = 36.0 \u00b5M). Kinetic studies revealed that KG and MG inhibited monophenolase activity of tyrosinase in a competitive manner. Docking simulations of KG and MG demonstrated favorable binding energies with amino acid residues of the active sites of tyrosinase. Our investigation of the structure-activity relationship of the fused benzofuran flavonoids (MG vs. AB) implicated the methyl cyclohexene ring moiety in tyrosinase inhibition. The enzyme substrate and relative structural analyses demonstrated that KG and MG from Morus sp. could be useful natural tyrosinase inhibitors in foods or cosmetics.Kuwanon G (KG) and benzofuran flavonoids such as mulberrofuran G (MG) and albanol B (AB) isolated from  These steps are crucial for skin protection against UV damage. Functional deficiencies of this pathway can result in serious dermatological diseases. Mushroom tyrosinase is a commercially available tyrosinase that has high homology with mammalian tyrosinase. Tyrosinase inhibitors are vital in the food industry [Tyrosinase is a ubiquitous enzyme in micro-organisms, animals, and plants. The oxidation of industry .Morus L. of the family Moraceae, are widely distributed. Fourteen species have been reported and classified by Zeng et al. [Bombyx mori L.), are used in Chinese herbal tea [Morus species.Mulberry species, genus g et al.  and are rbal tea , and arerbal tea . The riprbal tea . The learbal tea . Mulberrrbal tea , cardioprbal tea , antifunrbal tea , antioxirbal tea , hepatoprbal tea , and cytrbal tea  have beeMorus exhibited antibacterial [Morus australis [Morus nigra [Morus australis was previously investigated as an anti-obesity [Morus nigra contains phenolic compounds, including oxyresveratrol and mulberroside A [The tyrosinase inhibitory activity of kuwanon G (KG) is unclear ,13, but acterial , antioxiacterial , and hepacterial  activitiacterial . Albanolacterial , antibacacterial , and antacterial  activitiustralis  and Moruus nigra . Morus a-obesity  and skin-obesity  agent. O-obesity  along wi-obesity , phenoli-obesity , and fla-obesity . Morus nroside A , with neroside A , antioxiroside A .Structure\u2013activity relationship (SAR) studies can assist in identifying active moieties for the development of novel drugs. For this, it is necessary to understand the reaction mechanism. Chao et al.  demonstrl-tyrosine and l-DOPA as a substrate, respectively [Morus species with tyrosinase for the first time.Our study mechanistically investigated the reason behind the conflicting tyrosinase inhibitory activity of KG through monophenolase and diphenolase inhibitory assays with ectively . In addil-tyrosine . AB was found to be inactive. KG displayed significant tyrosinase inhibitory activity with an IC50 value of 67.6 \u00b5M . MG showed an acceptable IC50 value of 105.57 \u00b5M, while AB was inactive. Likewise, 1-methyl-1-cyclohexene moiety itself was inactive regarding both assays (IC50 > 1000 \u00b5M). However, our result for inhibition of mushroom tyrosinase by KG was significantly different from that of Zheng et al. [Three compounds from Morus species  were testyrosine . MG had  67.6 \u00b5M . KG showg et al.  who demol-tyrosine, 1.0, 0.5, and 0.25 mM) along with the various concentrations of KG  and MG  were analyzed. The lines of the Dixon plot intersected the y-intercept, which indicated competitive inhibition of tyrosinase with iK values of 18.66 and 5.19, respectively, for KG and MG .Tyrosinase is a rate-limiting, copper-containing enzyme that controls the production of melanin in the human body, which can lead to a variety of skin disorders when overproduced . Many nal-tyrosine and l-DOPA oxidation assays to investigate the inhibition activity of these compounds against mushroom tyrosinase. l-Tyrosine and l-DOPA are consecutive substrates, inducers, and positive regulators of melanogenesis that can regulate melanocyte function through overlapping pathways [l-tyrosine oxidation was inhibited by MG (IC50 = 17.53 \u00b5M), which was consistent with our results. In addition to a positive trend, MG was two times more potent than the result obtained in our current study. Slight variation can be attributed to experimental-setup errors. KG showed potent inhibition towards l-tyrosine in our study (IC50 = 67.6 \u00b5M), despite being previously reported as inactive up to 200 \u00b5M [50 = 27.2 \u00b5M) via the l-DOPA pathway. These results are in close agreement with our present study (IC50 = 44.0 \u00b5M).KG, MG, and AB have recently attracted extensive research focus. We systematically investigated these three compounds as potential candidates against Alzheimer\u2019s disease . As a papathways . Differepathways  determino 200 \u00b5M . Our stuo 200 \u00b5M  also repl-tyrosine pathway. The respective double reciprocal Lineweaver\u2013Burk plot revealed the competitive inhibition mechanism of KG and MG, and that both compounds readily bound to the catalytic site. MG had a greater binding affinity than KG as indicated by the lower iK value of 5.93. For these types of inhibitors, a higher substrate concentration is needed to achieve 50% occupation of the active sites. Kinetic studies revealed that both compounds were competitive inhibitors, indicating that they bind to the enzyme-substrate complex or interact with a specific catalytic site of the enzyme.Molecular mechanistic studies confirmed the tyrosinase inhibition activity of KG and MG via the Morus species were indicative of tyrosinase inhibition activity. AB had a smaller docking score than kojic acid, but the lack of interactions with copper and peroxide ions and catalytic residues of tyrosinase limited the inhibitory activity. On the other hand, MG and KG interacted with two copper ions and peroxide ions via van der Waals interaction and H-bond, respectively. Multiple hydrophobic interactions were also observed in the protein-KG and \u2013MG complexes. Interestingly, the methyl cyclohexene moiety of KG and MG formed multiple hydrophobic interactions with enzyme residues, whereas the methyl benzene ring of AB weakly hydrophobic interacted with the Val248 residue of enzyme. This is the first report of tyrosinase inhibition activity of MG and KG supported by enzyme kinetic analysis and molecular docking simulation.Molecular docking studies model the interaction between a small molecule and a protein at an atomic level, which allows characterization of small molecule behavior in the binding site of target proteins and elucidation of fundamental biochemical processes. The binding interactions of MG, KG, and AB from Structure\u2013activity relationships might explain the variable activities observed in these flavonoids. We studied the roles of methyl benzene, resorcinol, isoprenyl, and 4-pyrone moieties, focusing on the methyl cyclohexene ring structure. KG is a prenylated flavonoid, while MG and AB are fused benzofurans with specific roles. Zheng et al.  demonstrSophora flavescens showed tyrosinase inhibitory activity that was attributed to the resorcinol moiety [In another case, previous studies have suggested that the presence of resorcinol moiety in prenylated flavonoids is responsible for the activity. Major compounds kurarinol and kuraridinol from l moiety . The resOur study suggested that the methyl cyclohexene ring was responsible for inhibitory action against tyrosinase, which is an established hypothesis. The recent evaluation of the essential oils \u03b1-pinene and \u03b1-terpineol for tyrosinase and melanogenesis inhibition demonstrated positive results . Methyl AB has a methyl benzene ring in place of methyl cyclohexene, which explains its inactivity. Benzene has a resonance structure that leads to this inactivity: a large activation energy is required to break the double bonds of unsaturated hydrocarbons. The three double bonds of benzene are delocalized in a pi-system, so a low electron density results in low reactivity. MG forms bonds readily via a methyl cyclohexene functional group instead of a benzene group, which explains the tyrosinase inhibitory activity. The methyl cyclohexene moiety in KG also demonstrated acceptable inhibitory activity. KG has a similar structure to kojic acid, which contains 4-pyrone and might also contribute to its inhibitory activity. Our study supports the functionality of methyl cyclohexene in tyrosinase inhibition.Morus alba by Kuk et al. [l-DOPA, kojic acid, and mushroom tyrosinase were purchased from Sigma-Aldrich Co. . l-Tyrosine was purchased from Jannssen Chimica . K2HPO4 was obtained from Junsei Chemical Co. Ltd. , and KH2PO was acquired from Yakuri Pure Chemicals Co. Ltd. . Reagent-grade chemicals and solvents were purchased from commercial sources. Ultra-pure water was used throughout the experiment.KG, MG, and AB were obtained from the root bark of k et al.  as previl-DOPA and l-tyrosine oxidization activity of KG, MG, and AB isolated from root bark of Morus alba in our previous study with a modified spectrophotometric method [l-tyrosine/l-DOPA solution, 50 mM potassium phosphate buffer (pH 6.5), and distilled water was 10:10:9. The reaction mixtures were incubated at 25 \u00b0C for 15 s. The absorbance of the mixture at 490 nm was measured using a microplate reader. The degree of inhibition of the sample was expressed as the concentration required for 50% inhibition (IC50). One unit (U) of enzymatic activity referred to the amount of an enzyme capable of converting 1 \u00b5M of the l-tyrosine/l-DOPA reaction mixture within 1 min at 280 nm under the specific conditions .We investigated the c method ,41. Briel-tyrosine  substrate concentration to determine the tyrosinase inhibitory mechanism. The reaction mixture consisted of a 20 \u00b5L test sample at the following test concentrations: 1.6, 8, and 16 \u00b5M for MG; 10, 50, and 100 \u00b5M for KG. Tyrosinase inhibition was determined at various concentrations of l-tyrosine substrate  in the absence or presence of the test compound concentrations  using Lineweaver-Burk double reciprocal plots that determine enzyme kinetic parameters including mK and maxV. The enzymatic procedures consisted of the previously described tyrosinase assay methods. The inhibition constant (iK) was determined from the interpretation of Dixon plots.We observed the sample activity by decreasing the l-tyrosine, kojic acid, and cinnamic acid were downloaded from PubChem Compound (NCBI), with compound CIDs of 5281667, 196583, 480819, 6057, 3840, and 444539, respectively. Kojic acid and cinnamic acid [The H-subunit (residues 2\u2013392) of deoxy-form mushroom tyrosinase protein was obtained from the RCSB Protein Data Bank (ID: 2Y9X) . Small mmic acid  were uset-test , and P < 0.05 was used as a statistically significant cut-off value. All results were expressed as the mean \u00b1 S.E.M. of triplicate experiments.Differences between the control group and the test group were determined using Student\u2019s To sum up, KG and MG have attracted much interest in recent years, which inspired us to explore their potential cosmetic applications through computational studies and structural elucidation. In addition, we have examined the inhibitory activities of KG, MG, and AB against both pathways of melanogenesis (monophenolase and diphenolase activities). Enzyme kinetic and computational assessment suggested that MG and KG were potent tyrosinase competitive inhibitors via H-bonding interaction with peroxide ion and van der Waals interaction with copper ions at the active site. This is the first report of MG, KG, and AB for enzyme kinetic mechanisms and molecular docking analysis to the best of our knowledge. The controversial inhibitory activity of KG has been resolved via kinetics and docking simulation studies, and the importance of the methyl cyclohexene functional group has been explored. Further studies of this moiety are necessary. Our results indicate that these active flavonoids might be promising candidates as cosmetic agents. However, further studies are needed to fully characterize the underlying mechanism responsible for the effects of KG and MG on murine or mammalian cell based assays."}
+{"text": "EM) and central memory T cells (TCM) from unprimed donors have decreased ability to induce graft-vs-host disease (GVHD). Allospecific TEM from primed donors do not mediate GVHD. However, the potential of alloreactive TCM to induce GVHD is not clear. In this study, we sought to answer this question using a novel GVHD model induced by T cell receptor (TCR) transgenic OT-II T cells. Separated from OT-II mice immunized with OVA protein 8 weeks earlier, the allospecific CD44high TCM were able to mediate skin graft rejection after transfer to naive mice, yet had dramatically decreased ability to induce GVHD. We also found that these allospecific CD44high TCM persisted in GVHD target organs for more than 30 days post-transplantation, while the expansion of these cells was dramatically decreased during GVHD, suggesting an anergic or exhausted state. These observations provide insights into how allospecific CD4+ TCM respond to alloantigen during GVHD and underscore the fundamental difference of alloresponses mediated by allospecific TCM in graft rejection and GVHD settings.Data from both animal models and humans have demonstrated that effector memory T cells (T On the contrary, the memory T cells, including effector memory T cells  and central memory T cells  from either untreated or allo-antigen primed donors, do not cause GVHD or cause only minor GVHD after transplantation (CM that express high level of CD44 do not induce GVHD (Graft-vs.-host disease (GVHD) is a major complication of allogeneic hematopoietic stem cell transplantation caused by alloreactive donor T cells . After bantation \u20136. Speciuce GVHD .EM in GVHD (\u2212/\u2212 mice following by in vivo priming with splenocytes from CB6F1 (H2b/I-E+ strain), TEM cells from the primed animals maintained the memory function to mediate skin graft rejection, but did not mediate GVHD when transplanted into lethally irradiated CB6F1 hosts. However, allospecific TCM population could not be generated in this model. To study the potential of alloreactive TCM to induce GVHD, we utilized a novel GVHD model induced by T cell receptor (TCR) transgenic OT-II T cells. Using this model, we were able to generate antigen-specific TCM by immunizing donor mice directly and further demonstrated that these cells mediated secondary skin graft rejection while did not induce GVHD.It has previously been reported that common virus specific memory T cells including EBV-specific and CMV-specific memory T cells do not GVHD in humans \u201311. Howe in GVHD . By tran+ mice and Luciferase+ mice  to generate OT-II+ Luciferase+ GFP+ triple positive mice. For all the strains, both female and male mice were used in this study. The donor mice were primed at 6\u20138 weeks old. The recipient mice were between 7 and 16 weeks old at the time of transplantation. All animal care and experimental procedures were approved by National Institute of Health and Duke University Institutional Animal Care and Use Committee.C57BL/6 mice were purchased from The Jackson Laboratory . B6.Cg-Tg(TcraTcrb)425Cbn/J (OT-II) mice and C57BL/6-Tg(CAG-OVA)916Jen/J (OVA) mice  were purin vivo, OT-II mice between the age of 6\u20138 weeks were immunized with OVA protein  emulsified in complete Freund's adjuvant  i.p. at 100 ug/mouse  bone marrow donors. T cells were depleted from bone marrow using anti-CD90.2 antibody and complement as previously published. In brief, bone marrow cells were flushed out from the long bones of donor mice and strained through a 70 \u03bcm cell strainer . Cells were then resuspended in cytotoxicity medium, incubated with anti-CD90.2 monoclonal antibody  at 4\u00b0C for 1 h. The cells were washed once and then resuspended in cytotoxicity medium containing 1:10 Low-Tox-M Rabbit Complement . The cells were then incubated at 37\u00b0C for 60 min and washed twice before use.N, 90~95% for TEM with 2\u20133% TCM contamination, and 86~92% for TCM with 2~9% TEM contamination and 1\u20132% TN contamination.OT-II mice primed for 8 weeks were used as T-cell donors. Purified T cells were separated from splenocytes using mouse Pan T Cell Isolation Kit II . The purified T cells were then stained with APC\u2013conjugated anti-CD62L (clone MEL-14), PE-conjugated anti-CD4 (clone CT-CD4), PerCy5.5-conjugated anti-CD44 (clone IM7) from BD PharmMingen , and sorted into different T-cell subsets according to 5 irradiated (20Gy) OVA splenocytes in a final volume of 200 \u03bcl. After incubation at 37\u00b0C in 5% CO2 for a specified period as indicated, cultures were pulsed with 3H-thymidine (1\u03bcCi [0.037MBq]/well). Cells were harvested after another 16 h of incubation, and counted in a MicroBeta Trilux liquid scintillation counter . Triplicate cultures were set up for each cell population tested.The proliferation assay was performed as described previously . Graded 7 TCD BM and different numbers of purified OT-II cells through tail vein. Survival and clinical scores of GVHD including body weight change, fur ruffling, skin changes, hunching posture, diarrhea, and activity were monitored daily. Moribund mice were sacrificed according to protocol approved by the Duke University Institutional Animal Care and Use Committee.OVA mice were lethally irradiated (10.5 Gy) using Cs irradiator and injected with 1 \u00d7 102 pieces, and kept on swab damped with cold PBS. The C57BL/6 recipient mice were anesthetized with isoflurane  with the right lumbar region shaved and sanitized with iodine solution followed by alcohol. A graft bed was prepared by removing an area of skin down to the level of the intrinsic muscle using fine scissors. The graft was fitted to the prepared bed, sutured with 5-0 surgical suture, and wrapped with an adhesive plastic bandage. The bandage was removed 4 days after surgery. Skin graft survival was assessed everyday by visual and caliper measuring. Rejection was defined as the first day when the entire epidermal surface area of the graft was <10% of original.The skin transplantation protocol was modified as previously published . In briein vivo imaging, mice were anesthetized with isoflurane and injected intraperitoneally with 50 mg/kg D-Luciferin  10 min before imaging with a Xenogen IVIS 100 imaging system  at maximum signal intensity using 5 min exposure time. Regions of Interest (ROIs) were drawn using Living Image 2.5 software .Mice were monitored for T-cell tracking once per week after bone marrow transplantation. For Single cell suspension of splenocytes were prepared as described before , 12. In t test, multiple t test, and multi-way ANOVA test were used. Level of significance was set at P < 0.05. Bar graphs represent mean \u00b1 SEM.Statistical analysis was performed using Prism GraphPad  and Excel . For survival studies, log-rank Mantel-Cox test was used. For MLR, body weight changes, GVHD score, and bioluminescent measurement, Student's N were sorted from OT-II mice as responding cells, and cocultured with 5 \u00d7 105 lethally irradiated OVA splenocytes as stimulators at graded ratio for different time period from 2 to 5 days. By analyzing the 3H-thymidine uptakes, it is shown that from T cells: OVA splenocytes 1:10 on, OT-II unprimed TN can be efficiently stimulated and proliferated , effector memory , and central memory  T cell subsets. Flow cytometry also confirmed that all the sorted antigen specific cells are GFP+  or trended to be shorter (P = 0.078) compared to that in TN recipients These data are consistent with the previous publications  and flow cytometry to track the antigen-specific T cell expansion  28 days . The GFPterwards . Althougytometry . AlthougEM and TCM by directly immunizing donor mice with alloantigens. These TEM and TCM were phenotypically the same as those isolated from polyclonal mice (EM and TCM separated from primed OT-II mice were functionally memory T cells because they rejected second-set skin grafts faster than TN did. By transplanting these different subsets of primed T cells into OVA mice, we proved that OT-II TN cells mediated the vigorous GVHD, while TEM did not cause GVHD in OVA mice. Although primed OT-II TCM resulted in some death within the first 3 weeks, the survival rate was still significantly higher than TN group, while the body weight recovery, GVHD score, and histological changes in the target organs were all similar to TEM recipients, indicating that TCM do not cause or cause very minor GVHD. These results are consistent with the previously published data demonstrating that alloreactive TEM and TCM would not cause GVHD (In our study, we successfully utilized the OT-II / OVA system to generate allo-specific Tnal mice . We furtuse GVHD , 12, 15.EM but also TCM against host antigen do not cause GVHD. Currently, naive T cells depletion and anti-virus memory T cells transfusion are under clinical trial for BMT patients to preserve T-cell anti-infection function while preventing GVHD (Our study has the important clinical significance in T-cell therapy in BMT patients. Antigen-specific T cells against host antigen are believed to be the major players in inducing GVHD. In our study, we demonstrated that not only antigen-specific Ting GVHD , 16\u201320. ing GVHD \u201324. Our One major difference between human and mouse memory T cells is that human memory T cells may contain true alloantigen specific T cells while those from normal mice do not. In humans, alloantigen specific memory T cells are generated when na\u00efve T cells are exposed to alloantigens during transfusion or pregnancy . Even thN cells that underwent vigorous proliferation in the first 3 weeks, OT-II TEM and TCM had very limited proliferation in the spleens after transplantation. This finding was supported by Dr. Brede's research, and further advanced his findings (EM cells, TCM had a more potent proliferation in the peripheral blood between Day 10 to Day 21. This may explain some of the GVHD related death in the first 3 weeks. When we further traced these cells, we found that even 30 days after the BMT, in the survivors' spleens we could still identify the retention of transplanted TCM and TEM cells. This result confirmed that the antigen specific memory T cells persist in the hosts after BMT but failed to induce GVHD, suggesting a potential status of T-cell exhaustion. Due to the limited number of T cells that we could recover from the recipient mice, we were unable to completely understand the mechanism by which alloreactive TCM could reject skin graft but could not induce GVHD, and why the existing alloreactive TCM remained for more than 30 days but did not induce GVHD. Hypothetically, we speculate that the fate of antigen-specific memory T cells would be different in the environment that encounters a small amount of removable antigens vs. the environment that is surrounded by a large amount of non-removable antigens. According to the previous research studying T-cell immunology in viral infections, (CM could reject skin graft, but could not induce GVHD later. We speculate that the long-term existing TCM would get exhausted to a specific non-removable alloantigen in an GVHD setting.We also further investigate the primed OT-II T cells proliferation and retention in the organ after transplantation. By using bioluminescent imaging, we proved that compared to Tfindings . Compareections,  memory Tections, , 28\u201331. d-specific, the CD4-mediated MHC-mismatched 3BBM74 model which is I-Abm12-specific and D10 model which is I-Ab-specific, and the CD4-mediated miHAg-mismatched TEa model, and the CD4-mediated TS1 TCR Tg model which recognize the S1 epitope of HA on the HA104 Tg mice (in vivo or in vitro, and transferred and expanded in RAG1\u2212/\u2212 mice. In these models, T-cell homeostasis is unpreventable, while the separation of TCM is hard to achieve due to continually CD44 expression. In Juchem et al. study, the use of in vitro immunized TS1 cells injecting into HA mice, which was a single-antigen TCR Tg model, reached similar conclusion about TEM with what we have seen in OT-II T-cell OVA host model (CM could not be well distinguished from the TN cells due to the continually CD44 expression, and mice receiving TCM had shown signs of GVHD. Thinking that the high potent of TN to cause GVHD, the different phenomenon of TCM in TS1-HA model and in our model may be caused by the very small number of TN contamination. In our OT-II-OVA model, due to a clearer separation of TCM subsets, we were able to focus on the CD44high expression population, and proved that antigen-specific TCM did not cause GVHD.In the previous studies, different TCR Tg T-cell of a single specificity models were used to study to alloreactive GVHD . These m Tg mice \u201336. In m+ T cells do not directly mediate GVT effect by themselves (CM are able to mediate direct GVT effect because they are CD4+ T cells. However, based on our finding that antigen-specific TCM was able to reject OVA-expressing skin graft (CM maintain at least indirect GVT activity.Based on the previous findings from Strober's group demonstrating that memory CD4emselves , we do nin graft  and the in graft , we beliin vivo, and further sorted into well differentiated subsets including TN, TEM, and TCM. This enables the study of single peptide antigen-specific subsets of T cells generated in a physical condition in vivo. Second, OVA is a commonly used labeled antigen on various cancer cell lines. OT-II / OVA GVHD model facilitates the study of the anti-tumor effect of different antigen-specific T-cell subsets in the GVHD model.The OT-II into OVA murine model that we provided is novel to study the antigen-specific memory T cells in murine GVHD model. There are two major advantages that contributed to the study. First, in OT-II mice, the memory T cells can be directly generated and expanded N cells can cause lethal GVHD, and 1,000 TCM cells can partially cause GVHD. The limitation of small number of cells becomes the obstacle for further cell tracing and cellular and molecular mechanism study. Secondly, the model is still preliminary. Further information about the exhaustion markers and functional assays of the cells injected would be more helpful to define the status and biological characteristics of the antigen specific memory T cells in GVHD.There are also some limitations and unanswered questions that need to be aware of. First, the antigen-specific model is very sensitive to T-cell number. Only 1,000 antigen-specific Tin vivo to generate memory T cells, and transplanting these T cells into OVA mice to induce GVHD. We have demonstrated that antigen-specific TEM and TCM model do not cause GVHD due to a decreased proliferation potency after BMT, but can exist in the hosts' organs for long possibly due to exhaustion.In conclusion, we have established a feasible antigen-specific TCR Tg GVHD model by immunizing OT-II mice BJC and WH designed the experiments. WH, WM, JJ performed the experiments. WH did the statistic analysis. WH, BJC, and NJC wrote the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."}
+{"text": "Two distinct (4+3)-nitroxy annulations between 1,5-enynes and anthranils have been developed to access tetrahydro-1H-benzo[b]azepine derivatives. H-benzo[b]azepine derivatives; the chemoselectivity varies with the types of alkynes. Terminal alkyne substrates deliver benzo[b]azepine derivatives via a novel skeletal rearrangement while internal 1,5-enynes afford products without a rearrangement process. To elucidate the mechanism of rearrangement, we performed 13C- and 2H-labeling experiments to identify the gold-containing isobenzofulvene intermediates, but their formation relies on the presence of anthranils.Two distinct (4+3)-nitroxy annulations between 1,5-enynes and anthranils have been developed to access tetrahydro-1 H-benzo[b]azepines bearing a hydroxyl (I\u2013IV) represent a family of privileged seven-membered azacycles,2 adrenergic agonists.I\u2013IV are generally long and tedious.b]azepine cores involves the development of stereoselective (4+3)-annulations between anthranils and all-carbon 1,3-dipoles (5Cyclic nitroxy species (N\u2013O) are widespread functionalities in numerous bioactive molecules and natural products.-dipoles , but onlelective +3-annulavia a N- or O-attack route, inevitably cleaving the N\u2013O bonds; selected examples are provided in eqn (2) and (3). We sought the first (4+3)-nitroxy annulations using alkyne-based 1,3-dipoles and anthranils. This work reports two distinct (4+3)-annulations of 1,5-enynes with anthranils; interestingly, the chemoselectivity varies with the alkynes. Terminal 1,5-enynes 1 (R = H) afford seven-membered nitroxy heterocycles 3via an unprecedented rearrangement in gold catalysis;5 derived from internal alkynes 4 are not skeletally rearranged, but are elaborated into various benzo[b]azepine frameworks -nitroxy annulations1a with anthranil 2a (1.2 equiv.) using various gold catalysts; the results are shown in 2 (L\u2032 = P(t-Bu)2(o-biphenyl), IPr, PPh3) afforded seven-membered nitroxy product 3a in 8\u201368% yield (entries 1\u20133), with P(t-Bu)2(o-biphenyl)AuCl/AgNTf2 being the most effective. To our delight, (PhO)3PAuCl/AgNTf2 increased the yield of the desired 3a up to 73% (entry 4); different silver salts as those in (PhO)3AuCl/AgX (X = SbF6 and OTf) delivered compound 3a in relatively low yields . With (PhO)3PAuCl/AgNTf2, the yields of compound 3a in different solvents were as follows: DCM (62%), acetonitrile (30%) and MeNO2 . AgNTf2 alone was completely inactive (entry 10). The molecular structure of compound 3a was characterized by X-ray diffraction2a, 1,5-enyne 1a was isomerized by a gold catalyst to afford 1\u2032-methylvinyl-1H-indene 1a\u2032, which was structurally unrelated to our target 3a. Anthranil 2a is obviously indispensable to enabling the (4+3)-annulations with structural rearrangement.We optimized the reactions of terminal 1,5-enyne H-indene 1a\u2032 was present as a byproduct in a minor proportion (5\u201315%). The annulations of anthranil 2a (1.2 equiv.) with terminal 1,5-enynes 1b\u20131d bearing various 4-phenyl substituents  proceeded smoothly to yield 3b\u20133d in 68\u201377% yields (entries 2\u20134). For their 5-phenyl analogues 1e\u20131g, the resulting annulation products 3e\u20133g  were obtained in 65\u201374% yields (entries 5\u20137). Variations of the olefin substituents as those in species 1h\u20131j  were still compatible with these new N\u2013O annulations to afford compounds 3h\u20133j in 55\u201367% yields (entries 8\u201310). We have also prepared a terminal alkyne such as 1-ethynyl-2-styrylbenzene 1k that gave a recovery yield (>95%) of two reactants under the standard conditions.Under these optimized conditions, we assess the generality of these new annulations with various terminal 1,5-enynes and anthranils. The results are provided in 2b\u20132f bearing various C(5)-substituents , yielding cyclic nitroxy species 3k\u20133o in 48\u201377% yields, with X\u2032 = OMe becoming less efficient (entries 11\u201315). Methoxy-containing anthranil 2e renders the gold catalyst less reactive because of its high basicity. This gold catalysis worked well with additional anthranils 2g and 2h bearing C(6)-substituents (Y\u2032 = Br and Me), yielding the desired 3p and 3q in 41% and 70% yields, respectively (entries 15 and 16). We also varied the C(3)-substituents of anthranils  to yield the desired 3r and 3s in 35% and 63% yields, respectively (entries 18 and 19). An effective range of alkynes and anthranils manifests the practicability of these new nitroxy annulations.We next examined anthranils 4a, but led to a distinct (4+3)-annulation reaction without a skeletal rearrangement. Among various gold catalysts, P(OPh)3AuCl/AgSbF6 was superior to its NTf2 catalyst analogue, delivering a nitroxy product 5a with respective yields of 78% and 68%; a molar ratio of 4a/2a = 1\u2009:\u20092.1 was the optimized condition. The molecular structure of 5a was inferred from its 5b analogue , delivering compounds 5b\u20135d and 5e\u20135g in 65\u201375% yields (entries 1\u20136). An X-ray diffraction study5b showing no skeletal rearrangement. 1,5-Enynes 4h and 4i bearing varied trisubstituted alkenes were also suitable for the reactions, affording the desired nitroxy species 5h and 5i in 71\u201372% yields (entries 7 and 8). When the alkyl substituents R were a cyclopropyl or CH2OTBS group, the corresponding compounds 5j and 5k were obtained in 56% and 53% yields, respectively (entries 9 and 10). We tested the reactions of various anthranils 2b\u20132f bearing various C(5)-substituents , giving the expected products 5l\u20135p in 55\u201375% yields with the methoxy substituent being less efficient (entries 11\u201315). For additional anthranils 2g and 2h bearing 6-substituents (Y\u2032 = Br and Me), the resulting products 5q and 5r were obtained in 48% and 68% yields, respectively (entries 16 and 17).We assess the scope of these nitroxy annulations with various internal 1,5-enynes 3a and 5a to manifest their synthetic utility. Treatment of species 3a with Zn in AcOH/MeOH/H2O6a in 89% yield while the reaction with Pd/H2 gave compound 6b efficiently. Alternatively, compound 5a was hydrolyzed with HCl/water to yield ketone derivative 7b that was convertible to 1-amino-5-ol 7c with Zn/AcOH reduction, and to the diol derivative 7d with Pd/H2 reduction. An imine reduction of species 5a was achieved with Pd/H2 to afford species 7a. Unexpectedly, Zn-reduction of species 5a in HOAc/MeOH/water led to a structural rearrangement to form compound 7e in 81% yield. The imine moiety of the initial 5a was incorporated into the structural skeleton of product 7e, but the mechanism is not clear at this stage. Molecular structures of compounds 7a and 7e were verified by X-ray diffraction.5a into 7e will be elucidated in a future study  scale\" fill=\"currentColor\" stroke=\"none\"><path d=\"M0 1440 l0 -80 1360 0 1360 0 0 80 0 80 -1360 0 -1360 0 0 -80z M0 960 l0 -80 1360 0 1360 0 0 80 0 80 -1360 0 -1360 0 0 -80z\"/></g></svg>C\u2013H carbon, and its resulting product 3a contained the 13C-content only at the alkyl C\u2013H carbon (eqn (6)). Isobenzofulvene species In 1 was unlikely to occur here although it was observed in a ruthenium-catalyzed cycloisomerization.2O, we found that the resulting 1-3ad contained deuterium (X = 0.29D) only at its alkenyl C\u2013H moiety (eqn (7)). Accordingly, gold-containing isobenzofulvene In 2 is compatible with these 13C and 2H-labeling experiments.Among the two nitroxy annulations, the mechanism for terminal 1,5-enynes 4 react with LAu+ to form cyclopropyl gold carbenes B (or B\u2032) in two resonance forms; exo-(4+3)-annulations of species B\u2032 with anthranils 2a likely yield gold-carbene species C that subsequently capture a second anthranil to yield products 5. This mechanism is essentially the same as that of their annulations with nitrosoarenes.B/B\u2032 is also likely to occur. Terminal 1,5-enyne 1a also generates cyclopropylgold carbene E because its cycloisomerization product 1a\u2032 is also a 1-vinylindene derivative. We envisage that the cyclopropyl C\u2013H proton of gold carbene E is acidic because of its proximity to the gold carbene functionality; the deprotonation with anthranil 2a generates cyclopropylidenylgold species F that undergoes a \u201cmethylenecyclopropane-trimethylenemethane\u201d rearrangement,In 2. An exo-(3+4)-annulation between fulvene In 2 and anthranil 2a affords the observed product 3a. The intermediacy of organogold species G is supported by 2H and 13C-labeling experiments.D \u2192 G. Four possible paths 1\u20134 are considered; Path 1 is our proposed mechanism in E from \u03c0-alkyne D has a low barrier of 9.1 kcal mol\u20131; the anion-promoted deprotonation of gold carbene E to form cyclopropylidenylgold species F is operable as the enthalpy cost is 16.9 kcal mol\u20131; the energy of species F is slightly higher than that of \u03c0-alkyne D by only 6.6 kcal mol\u20131. The remaining steps F \u2192 In 2 and In 2 \u2192 G are also operable as the transition states TS-F-In2 and TS-In2-G are close to \u03c0-alkyne D energy levels. One notable feature is that the enthalpy of transition state TS-F-ln2 is surprisingly smaller than that of species F by \u20130.3 kcal. This atypical case has similar precedents in the literature.TS-F-In2 has less zero-point vibration energy than F, due to the loss of one degree of freedom in the transition state. This also means that F \u2192 In2 is a barrierless process.Density functional theory calculations were then performed to investigate the feasibility for the key steps E and anthranil 2a. The reaction proceeds in a stepwise manner. As shown in 2a at gold carbene E produces species stepE by an endothermic process ; its activation energy is as high as 25.4 kcal mol\u20131. In the next step involving stepE \u2192 GH, the energy level of step-GHTS-E is higher than that of 1,5-enyne D by 18.1 kcal mol\u20131. We conclude that Path 2 is not as feasible as Path 1 according to We next examined the energy profiles in the (4+3) annulations (Path 2) between cyclopropyl gold carbenes E \u2192 In2), in contrast to a stepwise process in Path 1 (E \u2192 F \u2192 In2). Despite multiple attempts, we were unable to locate the transition state for the direct E \u2192 In2 step, suggesting that Path 3 probably does not exist. In Path 4, a ring opening takes place initially (E \u2192 In2-H), followed by deprotonation (In2-H\u2192In2). However, our calculations show that this pathway is unlikely to occur as we are unable to locate In2-H; all geometry optimizations lead to E.We also considered the remaining Paths 3 and 4, as depicted in b]azepines using 1,5-enynes, anthranils and a gold catalyst. Internal 1,5-enynes deliver these cyclic nitroxy species without skeletal rearrangement while their terminal alkyne analogues afford distinct annulation products with skeletal rearrangement. To elucidate the mechanism of this rearrangement, 2H and 13C-labeling experiments were performed to identify the intermediates of gold-containing isobenzofulvene species, the formation of which is dependent on the presence of anthranils.Before this work, Au- and Pt-catalyzed annulations of anthranils with alkynes typically produced azacyclic products that cleaved the N\u2013O bonds. To develop new (4+3)-annulations of alkyne-derived 1,3-dipolesThere are no conflicts of interest to declare.Supplementary informationClick here for additional data file.Crystal structure dataClick here for additional data file."}
+{"text": "The mechanical tests of the mussel threads show that both strength and extensibility of the threads decrease with increasing oxygen contents, but this property degradation can be restored upon removing the oxygen. The first\u2010principles calculations explain the change in iron coordination, which plays a key role in strengthening, degradation, and self\u2010healing of the polymer networks. The oxygen absorbs on metal ions, weakening the iron\u2010catecholate bonds in the cuticle and collagen core, but this process can be reversed by sea water. These findings can have important implications in the design of next\u2010generation bioinspired robust, highly extensible materials, and catalysis.Metal\u2010containing polymer networks are ubiquitous in biological systems, and their unique structures enable a variety of fascinating biological behaviors. Cuticle of mussel byssal threads, containing Fe\u2010catecholate complexes, shows remarkably high hardness, high extensibility, and self\u2010healing capability. Understanding strengthening and self\u2010healing mechanisms is essential for elucidating animal behaviors and rationally designing mussel\u2010inspired materials. Here, direct evidence of Fe 3+ and Fe2+ gradient distribution across the cuticle thickness of bypass thread is provided, which supports that the cuticle is a functionally graded material with high stiffness, extensibility, and self\u2010healing capacity. The oxygen absorbed on metal ions can weaken the iron\u2010catecholate bonds in the cuticle and collagen core, but cuticle can heal itself in sea water.Evidence of Fe Mytilus galloprovincialis), and high extensibility (\u2248100%).Nature\u2010evolved organisms have produced a variety of unique materials that possess extraordinary abilities or characteristics, such as high strength and toughness, self\u2010healing, efficient energy conversion, brilliant structural colors, intelligence, and so on, which provides inspiration for researchers to create a variety of advanced materials with similar properties.2+, for a linear crosslinking, and tris\u2010complex coordination, Fe3+, for 3D crosslinking), response to the environment , and functioning as sacrificial bonds to prevent catastrophic material failure.E < 50 MPa) while the cuticle shows hardness up to 1 GPa and stiffness up to 2 GPa.The extraordinary properties of the byssal threads stem from their unique hierarchical structures that combine collagenous cores surrounded by protective cuticle, mainly containing catecholic amino acid 3,4\u2010dihydroxyphenylalanine (DOPA) and iron, which form crosslinking iron\u2010catechol structures. Here, the iron is essential to the mechanical and self\u2010healing properties. It was reported that the cuticle showed twofold decrease in hardness after removing metals from it.2+ existing inside the cuticle, suggesting that the cuticle is a functionally graded material. The transition between Fe2+ and Fe3+ plays a key role in maintaining high stiffness, extensibility, and self\u2010healing of the cuticle. Our experiment combined with the first\u2010principles and finite element simulations demonstrated that oxygen might weaken and even break the Fe\u2010DOPA bonds, which led to the degradation of mussel threads, but the weakened and broken bonds in Fe\u2010DOPA complexes could be restored spontaneously in the presence of sea water. These findings provide a theoretical base of designing next\u2010generation mussel\u2010inspired coating surfaces which have high stiffness, extensibility, and self\u2010healing capacity.In this contribution, we have demonstrated the first direct evidence of FeFigureM. galloprovincials cuticle consists of biphasic granules in homogenous matrix. The granules have a size ranging between 0.5 and 2 \u00b5m and comprise around 50% of the cuticle volume  such that the mussel threads were tested in wet state with different oxygen contents in all the cases. Figure S7 in the Supporting Information shows the collection of all the stress\u2013strain curves measured under different environments. Like other biological materials, curves also have large variations in strength and extensibility, but the average stress\u2013strain curves Figure3A cleaThe fracture surfaces of the tested cuticles were examined with SEM. Figure FigureM. galloprovincials (hardness of 133 \u00b1 17.4 MPa and Young's modulus of 1.7 \u00b1 0.1 GPa).To understand the matrix\u2013granule bridging behavior of the cuticle, we measured the mechanical properties of the transition area between the granule and matrix using AFM. AFM has been proved as a reliable tool for imaging as well as measurements of the various physical properties of composite bio\u2010structures in the air and aqueous environments.2), the oxygen\u2010damaged cuticle is healed by removing the adsorbed oxygen, and the core is well protected from the crack attack in the cuticle.To fundamentally understand the mechanism of mechanical response of mussel threads to different environments, multiscale modeling was developed at the macro and molecular scales. First, the finite element modeling of whole byssal threads  was carried to determine the mechanical properties of the collagen core and cuticle coating . By fitting the mechanical behaviors of intact and cut byssal threads , it was found that Ogden hyperelastic model, which is widely used for soft biological materials,Figures2 could adsorb on (bis\u2010)Fe\u2010DOPA coordination complex (Fe2+DOPA2) weakening the Fe\u2010DOPA bonds. The change in Gibbs free energy for the oxygen adsorption was calculated, yielding 0.056 and \u22120.06 eV in a neutral and a weak acidic media, respectively . The nearly zero free energy change indicates that the reaction is thermodynamically possible. The O2 absorption changes the electronic/bonding structures of Fe2+\u2010DOPA2 , but our results showed that oxygen molecule could not adsorb on the surface of the DOPA. Therefore, DOPA chains will remain intact in the oxygen\u2010containing environments.To understand how the cuticle and collagen core degrade at the molecular scale in the oxygen\u2010containing environments, we further performed the first\u2010principles calculations, using the density function theory (DFT) methods, and determined the electronic structures and chemical reaction pathways under different environments . Although mytilus byssus contains roughly 25\u201330 different proteins, only one protein, mfp\u20101, is associated with certainty with byssal cuticles.3+DOPA3) is transformed into Fe2+DOPA2 through oxygen reduction reaction. In this transformation (Reaction (a) in Figure 2 would adsorb on a DOPA chain and break the bond with the iron through valence tautomerism, catalyzed by catechol dioxygenase enzymes that exist in the threads.3+DOPA3 transfers into Fe2+DOPA2. As an iron\u2010DOPA bond is broken, the 3D Fe3+DOPA3 crosslinking structures now become 2D Fe2+DOPA2 linear structures on the surface of the cuticle. However, the degradation process can only occur when a radical oxygen coupling O2 is generated by, e.g., light radiation.In oxygen\u2010containing environments, it is possible that (tris\u2010)Fe\u2010DOPA coordination complex (Fe2+DOPA2 since the change in Gibbs free energy of the oxygen dissociation is \u22120.14 eV in sea water (pH = 8.1) . Furthermore, a DOPA would react with Fe2+DOPA2 to form Fe3+DOPA3, with a change in Gibbs free energy of \u22120.47 eV in sea water, suggesting that the reaction could occur spontaneously. After this reaction, the damaged polymer chains are healed. The mechanical properties of the mussel threads are therefore recovered due to the healing. It should be noted that the first reaction is the key step in the healing process since sea water not only reduces the reaction energy but also promotes the electron transfer between the iron ions and the ions in the sea water. Therefore, sea water effectively protects the mussel threads from the oxygen attack, or promotes the healing of damage if any.In the aqueous environments, the damaged area of cuticle could be healed through oxygen evolution reaction (Reactions (c) and (d) in Figure 3+ were enriched on the thread surface  at 100% humidity environment condition, and the speed of all the tension tests was carried at 10 mm s\u22121 unless noted. While 100% humidity environment condition was kept, oxygen, air, and nitrogen were filled into the testing chamber before those tension tests. All the thread diameters and sizes were exampled before and after the tests. The hardness and Young's moduli values for the granule and matrix inside the cuticle were obtained by AFM nanoindentation test. The spring constant of the probe 5.4 N m\u22121 with a diameter of 2 nm was used . The indentation tests were carried out on cross\u2010sections of fresh threads with test specimens submerged in sea water.TEM Imaging: Threads were fixed in glutaraldehyde for 2 h (T = 4 \u00b0C), PBS  buffer rinsed two times, and then fixed in osmic acid for 2 h, followed by dehydration in acetone under standard protocols, then soaked in 1:1 pure acetone and Epon 812 epoxy resin mixed for 30 min for 2 h. Then sample was embedded in Spurrs resin  and microtome to produce thin sections of 100 nm, following standard protocols. Micrographs were obtained using a JEOL 123 TEM  operated at 80 kV.XANES Examination: XANES and near edge X\u2010ray absorption fine structure ( NEXAFS) spectra experiments were performed at the photoemission end\u2010station at beamline BL10B in the National Synchrotron Radiation Laboratory (NSRL) in Hefei, China. This beamline was connected to a bending magnet and equipped with three gratings that cover photon energies from 100 to 1000 eV with a typical photon flux of 1 \u00d7 1010 photons s\u22121 and a resolution (E/\u0394E) better than 1000 at 244 eV. The end station was comprised of four chambers including analysis chamber, preparation chamber, high pressure reactor, and load\u2010lock chamber. The base pressures were 2 \u00d7 10\u221210, 2 \u00d7 10\u221210, 5 \u00d7 10\u22129, and 5 \u00d7 10\u22129 mbar, respectively. The analysis chamber was connected to the beamline and equipped with a VG Scienta R3000 electron energy analyzer, a twin anode X\u2010ray source (Mg Ka and Al Ka), an UV light source, a rear\u2010view optics for low energy electron diffraction, and a high precision manipulator with four\u2010degree\u2010of\u2010freedom ). The preparation chamber was comprised of an Ar+ sputter ion gun, a quartz crystal microbalance, a quadrupole mass spectrometer (Pfeiffer QMS220), a high precision manipulator with four\u2010degree\u2010of\u2010freedom , and several evaporators. The high\u2010pressure reactor performed the in situ high pressure experiments under different atmosphere condition, which had a maximum range of 2 MPa and a highest temperature of 650 \u00b0C.\u221210 mbar. The resolving power of the grating was typically E/DE = 1000, and the photon flux was 1 \u00d7 10\u221210 photons s\u22121. Spectra were collected at energies from 700 to 740 eV in 0.2 eV energy steps. The NEXAFS raw data were normalized by a procedure consisting of several steps. First, the photon energy was calibrated from the 4f spectral peak of a freshly sputtered gold wafer. Then a line was substrated to set the pre\u2010edge to be zero. Finally, the spectra were normalized to yield an edge\u2010jump to one.The Fe XANES spectra were measured at the photoemission end\u2010station at beamline BL10B in the National Synchrotron Radiation Laboratory (NSRL) in Hefei, China. A bending magnet was connected to the beamline, which was equipped with three gratings covering photon energies from 100 to 1000 eV. In this experiment, the samples were kept in the total electron yield mode under an ultrahigh vacuum at 5 \u00d7 10E/\u0394E) better than 1000. The end\u2010station was composed of four chambers, i.e., analysis chamber, preparation chamber, quick sample load\u2010lock chamber, and high pressure reactor. The analysis chamber, with a base pressure of < 5 \u00d7 10\u221210 Torr, was connected to the beamline and equipped with a VG Scienta R3000 electron energy analyzer and a twin anode X\u2010ray source. The high\u2010pressure reactor housed a reaction cell where the samples could be treated with different gases up to 20 bar and simultaneously heated up to 650 \u00b0C. After the sample treatment, the reactor could be pumped down to high vacuum (<10\u22128 Torr) for sample transfer. In the current work, the sample was deposited on the system to analysis chamber for XPS measurement without exposing to air.In situ XPS measurements were performed at the photoemission end\u2010station at beamline BL10B in the National Synchrotron Radiation Laboratory (NSRL) in Hefei, China. Briefly, the beamline was connected to a bending magnet and covered the photon energies from 100 to 1000 eV with a resolving power (DFT Simulations: A series of models was developed to simulate the reactions of MFP\u2010Fe\u20106O and MFP\u2010Fe\u20104O with various oxygen ligands. DFT with Hubbard (DFT + U) was carried out using soft projector\u2010augmented wave pseudopotentials and the Perdew\u2013Burke\u2013Ernzenhof exchange correlation functional, as implemented in the VASP code. The plane wave kinetic energy had a high cut of energy of 550 eV throughout the computations and a vacuum spacing of at least 20 \u00c5 in the z\u2010direction. The k\u2010point setting of Brillioun zone was obtained by 3 \u00d7 3 \u00d7 3 grid generating meshes with their origin point at the gamma point. Moreover, all the spin\u2010polarized calculations were converged to 0.01 eV \u00c5\u22121 for all surfaces and the geometries.Confocal Laser Scanning Microscopy (CLSM) Imaging: Fresh mussel was cut and transferred to glycerine  and mounted using high\u2010precision cover slips. Then the autofluorescence of gecko setae was analyzed using the CLSM TCS SP8  equipped with an inverted microscope and two solid\u2010state lasers  and Argon laser. For the visualization, a 40 \u00d7 objective  was used.The authors declare no conflict of interest.Supporting InformationClick here for additional data file."}
+{"text": "Le cancer de la vulve est une affection n\u00e9oplasique rare, repr\u00e9sentant moins de 5% des cancers g\u00e9nitaux de la femme. L'objectif de ce travail est de d\u00e9crire le profil \u00e9pid\u00e9miologique, clinique, paraclinique, th\u00e9rapeutique et \u00e9volutif du cancer de la vulve chez la population de la r\u00e9gion de l'Oriental du Maroc, ceci \u00e0 travers une analyse r\u00e9trospective de toutes les patientes suivies pour un cancer de la vulve, de juin 2007 \u00e0 janvier 2014, et trait\u00e9es au sein de l'h\u00f4pital d'Oncologie du Centre Hospitalier Universitaire Mohamed VI de Oujda au Maroc. Notre analyse r\u00e9trospective a port\u00e9 sur 34 patientes, d'une m\u00e9diane d'\u00e2ge de 65,7 ans, dont 52,9% \u00e9taient des multipares. Le motif de consultation dominant \u00e9tait le prurit dans 94.1% des cas. La moyenne du d\u00e9lai de consultation \u00e9tait de 16 mois, allant de 2 mois \u00e0 8 ans. L'ignorance et la pudeur ont \u00e9t\u00e9 les causes majeures de ce retard diagnostique, puisque 73.5% des patientes avaient d\u00e9j\u00e0 une maladie localement avanc\u00e9e au diagnostic. Le traitement chirurgical a \u00e9t\u00e9 propos\u00e9 \u00e0 61.4% des cas, il a consist\u00e9 en une vulvectomie radicale avec un curage inguinal bilat\u00e9ral dans 68.5% des cas. Le recours \u00e0 la radioth\u00e9rapie adjuvante a \u00e9t\u00e9 indiqu\u00e9 chez 41.2% des cas, 5.9% des patientes ont b\u00e9n\u00e9fici\u00e9 d'une radioth\u00e9rapie n\u00e9o adjuvante, et 20,6% d'une radioth\u00e9rapie exclusive associ\u00e9e \u00e0 une chimioth\u00e9rapie concomitante. La chimioth\u00e9rapie palliative a \u00e9t\u00e9 propos\u00e9e pour 8.8% des patientes. Le taux de survie globale \u00e0 3 ans est \u00e0 65%, le taux de r\u00e9cidives locor\u00e9gionales ou \u00e0 distance est de 17.3% des cas. Les particularit\u00e9s culturelles et sociales des patientes de la r\u00e9gion de l'Oriental du Maroc, qui sont suivies pour un cancer de la vulve, sont des facteurs influen\u00e7ant le traitement et ses r\u00e9sultats. Des efforts de pr\u00e9vention et de sensibilisation suppl\u00e9mentaires sont \u00e0 mener afin de r\u00e9duire l'incidence des stades localement avanc\u00e9s, et de permettre un traitement curatif \u00e0 cette population. La richesse des modes de manifestation de ce cancer contraste avec le retard diagnostic chez la plupart des patientes de la r\u00e9gion de l'Oriental du Maroc, o\u00f9 la pudeur, souvent, les conduit \u00e0 consulter \u00e0 un stade avanc\u00e9, o\u00f9 les options th\u00e9rapeutiques sont moindres. L'objectif est de d\u00e9crire le profil \u00e9pid\u00e9miologique, clinique, histologique, th\u00e9rapeutique et \u00e9volutif des cancers de la vulve, chez la population orientale.Le cancer de la vulve constitue 5% des cancers gyn\u00e9cologiques, qui touchent essentiellement la femme \u00e2g\u00e9e. Le type histologique \u00e9pidermo\u00efde repr\u00e9sente 90% de l'ensemble de ces cancers, ou le Il s'agit d'une \u00e9tude r\u00e9trospective, men\u00e9e au Service de Radioth\u00e9rapie de l'h\u00f4pital d'Oncologie du Centre Hospitalier Universitaire Mohamed VI d\u2019Oujda, concernant une s\u00e9rie de 35 patientes suivies pour un cancer vulvaire, sur une p\u00e9riode de 7 ans de juin 2007 \u00e0 janvier 2014. L'ensemble des donn\u00e9es \u00e9pid\u00e9miologiques, cliniques, histologiques, th\u00e9rapeutiques et \u00e9volutives a \u00e9t\u00e9 recueilli des dossiers de l'archive de l'h\u00f4pital, et a \u00e9t\u00e9 analys\u00e9 via le logiciel SPSS, afin de comparer les r\u00e9sultats de chaque traitement.Nous avons recens\u00e9 34 patientes, dont l'\u00e2ge moyen \u00e9tait 65,7 ans (extr\u00eames 25-94 ans), elles \u00e9taient des multipares dans 52,9% des cas, et m\u00e9nopaus\u00e9es dans 82,3% des cas. 26,4% des patientes avaient des comorbidit\u00e9s. 7,6% des patientes ont pr\u00e9sent\u00e9 des l\u00e9sions pr\u00e9canc\u00e9reuses. Le d\u00e9lai moyen de consultation \u00e9tait de 16 mois (2 mois - 8 ans). Les signes cliniques \u00e9taient domin\u00e9s par le prurit dans 94,1% des cas, une masse vulvaire r\u00e9v\u00e9latrice dans 61,3% des cas, les douleurs pelviennes et des leucorrh\u00e9es dans 29.4% et 82.3% des cas, respectivement. Il s'agissait de l\u00e9sions multifocales dans 88,3% des cas, et d'aspect ulc\u00e9ro-bourgeonnant dans 62,8% des cas . La biopLe cancer de la vulve repr\u00e9sente 5% des cancers gyn\u00e9cologiques . Son incLe cancer \u00e9pidermo\u00efde de la vulve est consid\u00e9r\u00e9 comme un cancer radiosensible, l'irradiation peut se discuter en n\u00e9o adjuvant chez les patientes porteuses de tumeurs localement avanc\u00e9s  pour permettre un down-staging tumoral et une chirurgie d'ex\u00e9r\u00e8ses carcinologiques par la suite, la radioth\u00e9rapie adjuvante par contre est consid\u00e9r\u00e9e comme un standard th\u00e9rapeutique en cas d'envahissement ganglionnaire, en cas de limites tumorales ou de tumeur large de plus de 4cm, ou en cas de pr\u00e9sence d'embols vasculaires ou d'engainement p\u00e9ri-nerveux, apr\u00e8s une chirurgie d'ex\u00e9r\u00e8se. La radioth\u00e9rapie exclusive peut \u00eatre propos\u00e9e chez les patientes r\u00e9cus\u00e9es chirurgicalement pour des raisons m\u00e9dicales, en cas de tumeurs tr\u00e8s localement avanc\u00e9es jug\u00e9es irr\u00e9s\u00e9cables, ou en cas de r\u00e9cidive tumorale locale , 7. La cL'irradiation externe pelvienne est d\u00e9livr\u00e9e le plus souvent par une combinaison de photons et d'\u00e9lectrons au niveau de la vulve, des creux inguinaux et parfois des cha\u00eenes iliaques, en pr\u00e9 et postop\u00e9ratoire \u00e0 une dose de 45 \u00e0 50 Gy d\u00e9livr\u00e9s en 5 \u00e0 6 semaines. En cas de radioth\u00e9rapie exclusive, les doses d\u00e9livr\u00e9es sont comprises entre 60 et 70 Gy dans le volume vulvaire et entre 55 et 60 Gy dans les volumes inguino-cruraux. La radioth\u00e9rapie est r\u00e9alis\u00e9e dans notre s\u00e9rie d'une fa\u00e7on exclusive dans 7,4% des cas le plus souvent \u00e0 titre palliatif, en pr\u00e9op\u00e9ratoire dans 5,7% des cas dans les formes localement avanc\u00e9es inaccessibles \u00e0 un traitement chirurgical d'embl\u00e9e, en postop\u00e9ratoire dans 37.2% des cas. La radioth\u00e9rapie postop\u00e9ratoire est pratiqu\u00e9e devant des facteurs de risque de r\u00e9cidive locale: un volume tumoral sup\u00e9rieur \u00e0 4cm; une marge chirurgicale \u00e9troite moins de 8 mm), un envahissement vasculaire lymphatique profond et le statut ganglionnaire positif. Pour la radioth\u00e9rapie n\u00e9o adjuvante ou exclusive, indiqu\u00e9e devant les formes localement avanc\u00e9es, ou r\u00e9cidivantes, l'association avec une chimioth\u00e9rapie concomitante \u00e0 base de 5 fluroracile ou de cisplatine semble am\u00e9liorer les taux de r\u00e9s\u00e9cabilit\u00e9 et avec un taux de r\u00e9ponse compl\u00e8te \u00e0 40% chez quelques patientes rapport\u00e9es sur des s\u00e9ries r\u00e9trospectives. En adjuvant, la chimio-concomitante associ\u00e9e \u00e0 la radioth\u00e9rapie apr\u00e8s traitement chirurgical n'a pas encore \u00e9t\u00e9 \u00e9valu\u00e9e. Aucune patiente de la s\u00e9rie n'en a b\u00e9n\u00e9fici\u00e9  mm, un e. L'utiliLes particularit\u00e9s culturelles et sociales des patientes de la r\u00e9gion de l'Oriental du Maroc, qui sont suivies pour un cancer de la vulve, sont des facteurs influen\u00e7ant le traitement et ses r\u00e9sultats. Des efforts de pr\u00e9vention et de sensibilisation suppl\u00e9mentaires sont \u00e0 mener afin de r\u00e9duire l'incidence des stades localement avanc\u00e9es, et de permettre un traitement curatif \u00e0 cette population.Cancer de la vulve est d\u00e9sormais le cancer gyn\u00e9cologique qui a b\u00e9n\u00e9fici\u00e9 le moins des avanc\u00e9es th\u00e9rapeutiques qu'a connu le monde de canc\u00e9rologie moderne;Du fait de sa raret\u00e9, la litt\u00e9rature compte peu de publications la concernant, il s'agit surtout de revues r\u00e9trospectives.Notre travail constitue un penchant sur les th\u00e9rapies actuelles propos\u00e9es, par notre institution, pour les patientes porteuses d'un cancer de la vulve, ainsi que leurs r\u00e9sultats en termes de survie.Les auteurs ne d\u00e9clarent aucun conflit d'int\u00e9r\u00eats."}
+{"text": "The ophthalmic TL-NPs formulations were prepared by the bead mill method, which yielded a mean particle size of ~93 nm with or without MC (0.5\u20133%). Although the dispersibility of TL particles in ophthalmic formulations increased with the MC content, the diffusion behavior of TL particles in the dispersion medium decreased with MC content. In an in vivo study using rats, the TL content in the lacrimal fluid was enhanced with MC content in the ophthalmic TL-NPs formulations, and the optimum amount of MC (0.5\u20131.5%) enhanced the TL content in the cornea and conjunctiva, and an anti-inflammatory effect of TL in rats instilled with ophthalmic TL-NPs formulations was observed. On the other hand, excessive MC (3%) prevented the corneal uptake of TL-NPs after instillation, and the anti-inflammation effect of TL was lower than that of ophthalmic TL-NPs formulations with optimum MC (0.5\u20131.5%). In conclusion, we found that gel formulations of TL-NPs with 0.5 and 1.5% MC provided a prolonged pre-corneal and pre-conjunctival contact time of TL, and resulted in higher TL contents in the cornea and conjunctiva following instillation in comparison with TL-NPs with or without 3% MC. This is probably due to the balance between the higher residence time and faster diffusion of TL-NPs on the ocular surface. These findings provide significant information that can be used to design further studies aimed at developing ophthalmic nanomedicines.We previously developed ophthalmic formulations containing tranilast nanopartaicles , and found them to show high uptake into ocular tissues. In this study, we aimed to design an  Eye drops can be applied easily as therapy for ocular diseases, and their share is over 70% in the ophthalmic field. On the other hand, drug residence is impeded by tear turnover, the corneal surface, reflex blinking and nasolacrimal drainage  so that in situ gelling systems exist as solids before instillation, but change to a gel on the ocular surface (after instillation). An in situ gelling system seems to be a good candidate for prolonging the drug residence time and enhancing absorption into the ocular surface -formulations with TL-NPs]. In addition, we investigated the preventive effect of ophthalmic ISG-formulations with TL-NPs on inflammatory mediators, such as nitric oxide (NO) and tumor necrosis factor-\u03b1 (TNF-\u03b1), in lipopolysaccharide (LPS) induced-rat conjunctival inflammation.In the ophthalmic field,  surface . Three s surface  and pH-s surface , have be surface . Various systems . MC, a w systems , and und systems . In thisMale Wistar rats aged 7 weeks were purchased from Kiwa Laboratory Animals Co., Ltd. . Japanese albino rabbits (\u223c2.7 kg) were used to investigate the corneal toxicity of ophthalmic formulations. All animal experiments were performed in accordance with the guidelines of ARVO and Kindai University, and were approved on 1 April 2013 (project identification code KAPS-25-003) by the pharmacy committee for animal research of Kindai University. Inflammation in conjunctiva was induced by the injection (30 \u03bcL) of 0.2 mg/mL LPS in saline into the upper palpebral conjunctiva. After 30 min, traditional and ophthalmic TL-NPs formulations were instilled, and the eyes were kept open for 1 min after instillation to prevent washing out.TL powder and commercially available TL eye drops (TL-sol) were gifts from Kissei Pharmaceutical Co., Ltd.; 2-Hydroxypropyl-\u03b2-cyclodextrin (HP\u03b2CD) and type SM-4 methylcellulose (MC) were obtained from Nihon Shokuhin Kako Co., Ltd.  and Shin-Etsu Chemical Co., Ltd. , respectively. Benzalkonium chloride was purchased from Kanto Chemical Co., Inc. , and mannitol (D-mannitol), ethyl p-hydroxybenzoate and isoflurane were provided by Wako Pure Chemical Industries, Ltd. . All other chemicals used were of the highest purity commercially available.\u00ae column . The column was eluted at 35\u00b0C with 50 mM ammonium acetate and acetonitrile (80:20) at a flow rate of 0.25 mL/min. TL was detected at 230 nm.TM MAX-XP Ultracentrifuge  and HPLC method. For the measurement of solubility, non-solubilized TL-NPs were removed by centrifugation at 100,000 g, and the TL contents in the supernatants were measured. To measure the dispersibility of TL-NPs, ophthalmic TL-NPS formulations were stored for 3 months at 20\u00b0C, and samples were taken from 5 mm under the surface over time. The TL contents of the samples were determined by the HPLC method described above.The particle size of the TL powder was determined by a laser diffraction particle size analyzer Shimadzu SALD-7100  with the refractive index set at 1.60\u20130.10i. Particle size and number in the TL-NPs formulations were measured by a Dynamic Light Scattering QuantumDesign NANOSIGHT LM10  for 60 s at 405 nm. The viscosities at 20 and 37\u00b0C were analyzed by an Anton Paar MCR302 attached to a CP50-1 . The measurement was performed 10 times, and the mean was used in this study. The measurement conditions were as follows: measurement time 2 s, interval 1 s, shear rate 90\u2013100 rpm/s. The atomic force microscopic (AFM) image created by combining a phase and height image was provided by a Shimadzu SPM-9700 . A Nihon Rufuto micro-electrophoresis zeta potential analyzer model 502 was used to determine the zeta potential , and the crystal form was obtained using a Rigaku MiniFlex II . The samples for powder X-ray diffraction (XRD) were lyophilized and pulverized with the diffraction angles set from 5\u00b0 to 90\u00b0, and measured at a scanning rate of 10\u00b0/min. TL solubility was measured using a Beckman OptimaA methacrylate cell equipped with a 0.22 \u03bcm-pore membrane filter was used . One sidRats were instilled with ophthalmic TL-NPs formulations, and euthanized by injection of a lethal dose of sodium pentobarbital 30, 60, and 150 min after instillation. The lacrimal fluid was collected using schirmer tear test strips. Blood was collected from the vena cava, and centrifuged to provide serum. The corneas and conjunctiva were excised. TL was extracted from the lacrimal fluid in the schirmer tear test strips, blood (serum), cornea and conjunctiva in methanol, and centrifuged at 20,400 g for 20 min at 4\u00b0C. The supernatants were used as samples. TL contents were measured by the HPLC method described above. Protein levels in the samples were determined using a Bio-Rad Protein Assay Kit , and the TL levels in the corneas and conjunctiva are expressed as nmol/mg protein.Ophthalmic TL-NPs formulations (30 \u03bcL) were repetitive instilled to rabbits twice a day (9:00 and 19:00) for 1 month. After that, 30 \u03bcL of 1% fluorescein was instilled to stain the wound area, and the wound area were monitored by a Topcon TRC-50X .EB was used to monitor vascular protein leakage. The EB (10 mg/kg) was injected to femoral vein 5 min before the injection of LPS. When the experiment was finished , the rats were euthanized by injecting a lethal dose of pentobarbital, the blood was removed by perfusion with cold saline, and the conjunctiva were excised. The conjuntival tissues were homogenized in 1M KOH, and incubated for 24 h at 37\u00b0C. After that, the EB was extracted by incubation in 0.2 M phosphoric acid and acetone (5:13) for 2 h, after which the samples were centrifuged at 400 g for 15 min at 4\u00b0C, and the supernatants were used for measurements. EB exudation was determined from the absorbance at 620 nm, and is expressed as Abs/g weight of conjunctival tissue.2\u2013 and NO3\u2013 levels were detected at 540 nm. In this study, NO amounts represent the total NO metabolite level, which is the sum of the NO2\u2013 and NO3\u2013 levels.The conjunctiva were excised from rats after euthanasia were homogenized in saline on ice, and centrifuged at 20,400 g for 20 min at 4\u00b0C. The supernatants were filtrated by a concentric microdialysis probe , and perfused to Eicom ENO-20  with Ringer\u2019s solution at a constant flow rate of 2 \u03bcL/min. The samples were mixed Griess reagent in ENO-20, and the NORats were euthanized by injecting a lethal dose of pentobarbital, and the conjunctiva were excised and homogenized in saline on ice. The homogenates were centrifuged at 20,400 g for 20 min at 4\u00b0C, and the supernatants were used for the measurement of TNF-\u03b1 levels. TNF-\u03b1 levels were measured using a Rat TNF-\u03b1 Quantikine ELISA Kit  according to the manufacturer\u2019s instructions. The TNF-\u03b1 levels are expressed as pg/mg protein.P-value of 0.05 was chosen as the significance level (P < 0.05), and Student\u2019s t-test and one-way analysis of variance (ANOVA) followed by Dunnett\u2019s multiple comparison were used for statistical analysis of groups. The sample numbers (n) are shown in the figure legends, and the data are expressed as mean \u00b1 standard error of the mean (SEM).A minimum First, we designed formulations containing TL-NPs using the bead mill method, and evaluated their characteristic parameters. The changes in viscosity affect the diffusion of TL-NPs in dispersion medium. Therefore, we investigated the relationship between viscosity and diffusion in the ophthalmic TL-NPs formulations. In via energy-dependent endocytosis pathways  have been shown to exert various anti-inflammatory actions in experimental studies , and arereatment , while ireatment . Even threatment . These rreatment . The TL reatment . Thereforeatment .in situ gelling systems have been designed to prolong the pre-corneal contact time of ophthalmic formulations. Therefore, we attempted to prepare in situ gels incorporating HP\u03b2CD-coated NPs , and evaluate the effect of MC addition on the gel characteristics. Although the solubility, zeta potential, particle size, and nanoparticle number for TL in the ophthalmic formulations were similar with or without MC, the dispersibility was enhanced by MC with no aggregation or precipitation observed for 3 months  was 0.5\u20131.5% in ophthalmic ISG-formulations with NPs. These findings provide significant information that can be used to design further studies aimed at developing ophthalmic nanomedicines.In conclusion, we succeeded in preparing ophthalmic ISG-formulations with TL-NPs, and found that The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation, to any qualified researcher.The animal study was reviewed and approved by the Pharmacy Committee for Animal Research at Kindai University .NN conceived and designed the study and wrote the manuscript. NN and MM performed the experiments for the formulation and evaluation of nanoparticles, and analyzed the data. SD and HO performed the drug behavior experiments using rats. SD, HS, and NY performed the experiments to test the therapeutic effect on inflammation. All authors significantly contributed to the conception and design of the study, and to the interpretation of the data.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."}
+{"text": "A major goal in diseases is identifying a potential therapeutic agent that is cost-effective and can remedy some, if not all, disease symptoms. In Alzheimer\u2019s disease (AD), aggregation of hyperphosphorylated tau protein is one of the neuropathological hallmarks, and Tau pathology correlates better with cognitive impairments in AD patients than amyloid-\u03b2 load, supporting a key role of tau-related mechanisms. Selenium is a non-metallic trace element that is incorporated in the brain into selenoproteins. Chronic treatment with sodium selenate, a non-toxic selenium compound, was recently reported to rescue behavioral phenotypes in tau mouse models. Here, we focused on the effects of chronic selenate application on synaptic transmission and synaptic plasticity in THY-Tau22 mice, a transgenic animal model of tauopathies. Three months with a supplement of sodium selenate in the drinking water (12 \u03bcg/ml) restored not only impaired neurocognitive functions but also rescued long-term depression (LTD), a major form of synaptic plasticity. Furthermore, selenate reduced the inactive demethylated catalytic subunit of protein phosphatase 2A (PP2A) in THY-Tau22 without affecting total PP2A.Our study provides evidence that chronic dietary selenate rescues functional synaptic deficits of tauopathy and identifies activation of PP2A as the putative mechanism. N-glycosylation, and truncation) offers multiple targets for therapy development  is a progressive neurodegenerative disorder affecting predominantly brain regions that are required for information processing and memory such as hippocampus and cortex  sibling controls were used in the present study. THY-Tau22 and WT littermate mice were generated as described previously  and Passive Avoidance] was performed successively as depicted in 2) that were positioned between three infrared photo beams. Beam crossings that represented locomotor cage activity were registered for each 30-min interval during a 23-h recording period, using an interfaced PC  documented no motor deficits for THY-Tau22 , the latency of the first corner and center approach, as well as time spent, and velocity in corners and center were measured .Spatial and reference memory abilities were examined in the standard hidden-platform acquisition and retention version of the MWM. Training consisted of five consecutive days of four trials (120 s each) from different starting positions. The time required to locate the hidden escape platform (escape latency in seconds), distance traveled (path length in centimeter), and swimming speed (velocity in cm/s) were recorded . After 2 days of rest, a probe trial with the platform removed was performed for 100 s. The time (s) the animals spent in each of the four quadrants, and the frequency the mice swam over the former platform position (n) were measured . Animals were dark-adapted for 30 min and then placed in the small illuminated compartment. After 5 s, the sliding door to the dark compartment was opened and the entry latency was recorded. When mice entered the dark compartment, a mild foot shock  was delivered. Retention was tested 24 h later according to the same procedure, excepting that animals did not receive a shock. The entry latencies were recorded with a cut-off of 300 s.2/5% CO2). ACSF consisted of (in mM): 124 NaCl, 4.9 KCl, 24.6 NaHCO3, 1.20, KH2PO4, 2.0 CaCl2, 2.0 MgCl2, 10.0 glucose, pH 7.4. Transverse slices (400 \u03bcm thick) were prepared from the dorsal area of the right hippocampus with a tissue chopper and placed into a submerged-type chamber, where they were kept at 32\u00b0C and continuously perfused with ACSF at a flow-rate of 2.2 ml/min. After about 90 min incubation, one slice was arbitrarily selected and a lacquer-coated monopolar stainless steel or tungsten electrode was placed in CA1 stratum radiatum for stimulation in constant current mode. For the recording of field excitatory postsynaptic potentials (fEPSPs), a glass electrode  was placed in the stratum radiatum. The time course of the field EPSP was measured as the descending slope function for all sets of experiments. After a further hour of incubation, input/output curves were established and the stimulation strength was adjusted to elicit an fEPSP-slope of 35% of the maximum and was kept constant throughout the experiment. Paired pulse facilitation was investigated by applying two pulses in rapid succession  at 120 s intervals. During baseline recording, three single stimuli  were measured every 5 min and averaged. LTD was induced by low-frequency stimulation (LFS) consisting of 1,200 pulses at 2 Hz (0.2 ms pulse-width) and repeated three times with a 10-min interval between completion of one LFS-train and the start of the successive one  artificial cerebrospinal fluid (ACSF), oxygen saturated with carbogen  and GraphPad Prism 7 . Unless otherwise stated, all statistics were done with Two-Way ANOVA or repeated measures ANOVA (RM-ANOVA). For single between-group comparisons, F = 3.258, p = 0.035). Post hoc Dunnett tests confirmed that only vehicle-treated THY-Tau22 mice (henceforth referred to as Tg mice for the remainder of the manuscript) demonstrated a significantly different level of exploratory activity compared with age-matched WT mice  = 6.545, p = 0.002; post hoc Dunnett test vs. vehicle-treated Tg mice, p = 0.001). Chronic treatment with sodium selenate corrected for the increased initial and nocturnal activity of THY-Tau22 mice.When cage activity was monitored in the current study, we found a difference between groups  = 0.747, p > 0.05; data not shown). Previous investigations of the ambulatory activity of 10-month-old THY-Tau22 mice had demonstrated an overall increased activity in Tg mice  = 7.623, p = 0.001; post hoc Dunnett test p = 0.06). Time to approach the center of the arena and number of exploratory rearings in the center were different between groups  = 5.700, p = 0.03 for latency to the center and F = 3.723, p = 0.021 for rearing in the center). Non-treated Tg mice stayed longer away from and displayed less exploratory rearings in the center as compared to the vehicle-treated Tg animals  = 44.542, p = 0.0001 for escape latency and F = 43.466, p = 0.0001 for distance traveled). Post hoc Dunnett tests revealed that spatial acquisition in vehicle-treated Tg mice at 12\u201313 months of age was severely impaired compared with the other groups . This impairment was not due to motor deficits because swim speed did not differ across groups (p > 0.05 data not shown). Noteworthy, there was a pronounced and significant improvement in spatial learning of selenate-treated Tg mice which was comparable with the WT-groups. The comparison of vehicle- with selenate-treated Tg mice confirmed the rescue effect of chronic selenate treatment because the latter was statistically indistinguishable from the two WT groups in terms of escape latency and path length  = 9.775, p = 0.004). However, there was also a significant effect of group detected for the opposite quadrant  = 15.778, p < 0.001), and interaction effect  = 27.388, p < 0.001). Dunnett tests indicated that vehicle-treated THY-Tau22 failed to develop a preference for the target quadrant in contrast to the other three groups . Analysis of the latency to cross the former platform position revealed a trend towards poorer spatial reference memory in the vehicle-treated Tg mice compared to the other groups  = 3.893, p = 0.057).We assessed spatial learning and memory in the established hippocampus-dependent water maze paradigm. Looking at both the time to locate the hidden platform and the path length traveled to reach it, we found significant differences between the groups  = 32.348, p = 0.0001). Thus, the two WT-groups exhibited clear memory recall of the shock experienced 24 h earlier, whereas vehicle-treated Tg mice failed to remember as indicated by the short-latency to enter the dark compartment. Notably, in selenate-treated Tg animals, retention memory was restored to WT levels, resulting in a highly significant effect of treatment (post hoc Dunnett tests for vehicle-treated Tg compared to the other three groups p = 0.0001).Finally, in passive avoidance learning, a one-trial learning test, we found differences between the groups  = 3.389, p < 0.029) and post hoc Dunnett test revealed significant differences between selenate-treated WT and Tg mice (p \u2264 0.020). We then studied paired-pulse responses, a form of presynaptically mediated short-term plasticity. Tg mice displayed rather high paired-pulse ratios at short inter-pulse intervals of 10 ms and 20 ms independent of whether or not the animals had received selenate . Differences of similar size occurred also when the vehicle groups were compared, but the higher variability prevented statistical significance. While there were no significant differences at interpulse intervals of 50 ms and 100 ms, treatment with selenate led to significantly increased values in WT mice at 200 ms , the depression could not be maintained . Statistical analysis of LTD of all groups after the induction (60\u2013240 min) with RM-ANOVA plus Tukey\u2019s post hoc comparison, revealed a significant effect of group  = 6.843), p = 0.001 and significant differences between vehicle-treated Tg animals and all other groups . While the chronic oral application of sodium selenate had virtually no effect on WT mice, it completely rescued the L-LTD in Tg animals  at 2 Hz was successful in inducing late-phase LTD (L-LTD) in young and middle-aged mice  and selenate-treated Tg animals (n = 3) with AT8, a human tau pathological marker of S202/T205 .Histological examination of sections from vehicle-treated  was obtained.Since several reports have documented a downregulation of PP2Ac in murine models of tauopathies  expression and decreased APP cleavage  by selenate  at the 50 ms inter-pulse interval were found between treated and vehicle-treated Tg groups. Intriguingly, in WT mice selenate enhanced PPR at an interpulse interval of 200 ms, pointing to selenate-mediated effects on GABAIn Tg mice treated with selenate, we observed a restoration of LTD comparable with WT levels while selenate did not affect WT synaptic plasticity. This LTD rescue \u201cphenomenon\u201d was similar to the one that we have previously documented in response to a transient bath-application of selenate, which notably overcame okadaic acid inhibition (Ahmed et al., Given the current state of knowledge, it is rather difficult to identify a certain mechanism that is causally responsible for the selenate-mediated rescue effect on synaptic plasticity and cognition. Selenate, has been used as an anti-cancer agent for many years (Corcoran et al., In conclusion, we demonstrate for the first time that a 3-month treatment with dietary selenate rescues deficits in synaptic plasticity as well as learning and memory in Tg mice at an advanced stage of tauopathy, which led already to severe functional deficits at the synaptic, behavioral, and cognitive level. The completely recovered LTD demonstrates a rescue of the deleterious effects of progressive tauopathy at the level of the synapse, the main site of functional deterioration that leads to cognitive decline. This is most likely due to a reduction in the demethylated, catalytically inactive catalytic subunit PP2Ac, which shifts the equilibrium in favor of the methylated PP2Ac and thus to a higher PP2A activity. Consequently, this leads to a decrease in hyperphosphorylated, but not misconformed Tau as supported by the significant reduction in AT-8 but not in AT-100 immunostaining. Together with the clear rescuing effects on synaptic plasticity and cognitive measures in our study, this reinforces the idea that the detrimental impact of pathological Tau towards plasticity and memory is likely related to non-aggregated soluble Tau species and recommends selenate as a candidate compound for human Tau-targeting clinical trials.The raw data supporting the conclusions of this article will be made available by the authors upon reasonable request.The animal study was reviewed and approved by the Animal experiments committee of the University of Leuven, Belgium and were carried out in accordance with the European Directive 2010/63/EU.TA developed the idea for these studies. AVDJ and RD\u2019H designed the behavioral experiments which were conducted by AVDJ. The electrophysiological study was designed by TA and DBa, the biochemical and histological assays by DBl and LB. TA performed the electrophysiological measurements, DBl the biochemical and histological assays. AVDJ and DBa analyzed the behavioral data, TA, and DBa the electrophysiological experiments and DBl and RC the biochemical and histological assays with input from LB. DBa, TA, and AVDJ wrote the article with contributions from DBl and LB.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."}
+{"text": "A clear understanding of the macro-level contexts in which education impacts health is integral to improving national health administration and policy. In this research, we use a visual analytic approach to explore the association between education and health over a 20-year period for countries around the world.Using empirical data from the OECD and the World Bank for 26 OECD countries for the years 1995\u20132015, we identify patterns/associations between education and health indicators. By incorporating pre- and post-educational attainment indicators, we highlight the dual role of education as both a driver of opportunity as well as of inequality.Adults with higher educational attainment have better health and lifespans compared to their less-educated peers. We highlight that tertiary education, particularly, is critical in influencing infant mortality, life expectancy, child vaccination, and enrollment rates. In addition, an economy needs to consider potential years of life lost  as a measure of health quality.We bring to light the health disparities across countries and suggest implications for governments to target educational interventions that can reduce inequalities and improve health. Our country-level findings on NEET  rates offer implications for economies to address a broad array of vulnerabilities ranging from unemployment, school life expectancy, and labor market discouragement. The health effects of education are at the grass roots-creating better overall self-awareness on personal health and making healthcare more accessible. Is education generally associated with good health? There is a growing body of research that has been exploring the influence of education on health. Even in highly developed countries like the United States, it has been observed that adults with lower educational attainment suffer from poor health when compared to other populations . This paThe avenues through which education affects health are complex and interwoven. For one, at the very outset, the distribution and content of education changes over time . Second,Research has proposed that the relationship between education and health is attributable to three general classes of mediators: economic; social, psychological, and interpersonal; and behavioral health . EconomiWhile the positive association between education and health has been established, the explanations for this association are not . People On a universal scale, it has been accepted that several social factors outside the realm of healthcare influence the health outcomes . The difEducation enables people to develop a broad range of skills and traits  that predispose them towards improved health outcomes , ultimatThere is an iterative relationship between education and health. While poor education is associated with poor health due to income, resources, healthy behaviors, healthy neighborhood, and other socioeconomic factors, poor health, in turn, is associated with educational setbacks and interference with schooling through difficulties with learning disabilities, absenteeism, or cognitive disorders . EducatiTo inform future education and health policies effectively, one needs to observe and analyze the opportunities that education generates during the early life span of individuals. This necessitates the adoption of some fundamental premises in research. Research must go beyond pure educational attainment and consider the associated effects preceding and succeeding such attainment. Research should consider the variations brought about by the education\u2013health association across place and time, including the drivers that influence such variations .In the current research, we analyze the association between education and health indicators for various countries using empirical data from reliable sources such as the Organization for Economic Cooperation and Development (OECD) and World Bank. While many studies explore the relationship between education and health at a conceptual level, we deploy an empirical approach in investigating the patterns and relationships between the two sets of indicators. In addition, for the educational indicators, we not only incorporate the level of educational attainment, but also look at the potential socioeconomic benefits, such as enrollment rates  and school life expectancy . We investigate the influences of educational indicators on national health indicators of infant mortality, child vaccinations, life expectancy at birth, premature mortality arising from lack of educational attainment, employment and training, and the level of national health expenditure. Our research question is:What are some key influencers/drivers in the education-health relationship at a country level?The current study is important because policy makers have an increasing concern on national health issues and on policies that support it. The effect of education is at the root level\u2014creating better overall self-awareness on personal health and making healthcare more accessible. The paper is organized as follows: Section 2 discusses the background for the research. Section 3 discusses the research method; Section 4 offers the analysis and results; Section 5 provides a synthesis of the results and offers an integrated discussion; Section 6 contains the scope and limitations of the research; Section 7 offers conclusions with implications and directions for future research.Fundamental Cause Theory (FCT) [Research has traditionally drawn from three broad theoretical perspectives in conceptualizing the relationship between education and health. The majority of research over the past two decades has been grounded in the ry (FCT) , which pry (FCT) . Some ofry (FCT) .Human Capital Theory (HCT) that views it as a return on investment in the form of increased productivity [signaling or credentialing perspective [Education has also been conceptualized using the uctivity . Educatispective  - is adoIn the current research while we acknowledge the tenets of these theoretical perspectives, we incorporate the social mechanisms in education such as level of education, skills and abilities brought about by enrollment, school life expectancy, and the potential loss brought about by premature mortality. In this manner, we highlight the relevance of the social context in which the education and health domains are situated. We also study the dynamism of the mechanisms over countries and over time and incorporate the influences that precede and succeed educational attainment.We analyze country level education and health data from the OECD and World Bank for a period of 21\u2009years (1995\u20132015). Our variables include the education indicators of adult education level; enrollment rates at various educational levels; NEET  rates; school life expectancy; and the health indicators of infant mortality, child vaccination rates, deaths from cancer, life expectancy at birth, potential years of life lost and smoking rates , school life expectancy (tertiary), and NEET  rates, affected life expectancy at birth Fig. . Our expFigure We wanted to explore if the potential years of life lost rates are affected by tertiary enrollment rates and tertiary adult education levels Fig. .Fig. 4AThe two sets of box plots in Fig. We compared the performance of tertiary education level and enrollment rates with the child vaccination rates Fig.  to assesIn this analysis Fig. , we lookIn the realm of child health, we also looked at the infant mortality rates. We explored if infant mortality rates are associated with the NEET rates in different age groups Fig. .Fig. 6AFigure We looked to see if the distribution of population without employment, education, or training (NEET) in various categories of high, medium, and low impacted the rate of deaths from cancer Fig. . Our expThe three pie charts in Fig. It is interesting to note the relationship between health expenditure and adult education levels Fig. . We expeFigure Having explored the relationship between health expenditure and adult education, we then explored the relationship between health expenditure and NEET rates of different countries Fig. . We expeIn Fig. The distribution of enrollment rate (tertiary) and life expectancy of all the countries in the sample can give an idea of the current status of both education and health Fig. . We expeFigure We wanted to see if various levels of GDP per capita influence the levels of adult education and deaths from cancer in countries Fig. .Fig. 11Figure Infant mortality is an important indicator of a country\u2019s health status. Figure In Fig. We looked at the association between child vaccination rates and NEET rates in various countries Fig. . We expeFigure We compared the trend of average smoking rate for the years 1995\u2013201 for the continents in the sample Fig. .Fig. 14Figure We explored if adult education levels  are associated with deaths from cancer Fig.  such thaFigure We drilled down further into the correlation between the upper-secondary education level and deaths from cancer. Figure Figure We moved our focus to the trends of tertiary school life expectancy and health expenditure from 1995 to 2015 Fig.  to checkFigure Our results reveal how interlinked education and health can be. We show how a country can improve its health scenario by focusing on appropriate indicators of education. Countries with higher education levels are more likely to have better national health conditions. Among the adult education levels, tertiary education is the most critical indicator influencing healthcare in terms of infant mortality, life expectancy, child vaccination rates, and enrollment rates. Our results emphasize the role that education plays in the potential years of life lost, which is a measure that represents the average years a person would have lived had he/she not died prematurely. In addition to mortality rate, an economy needs to consider this indicator as a measure of health quality.Other educational indicators that are major drivers of health include school life expectancy, particularly at the tertiary level. In order to improve the school life expectancy of the population, governments should control the number of youths ending up unemployed, dropping out of school, and without skills or training (the NEET rate). Education allows people to gain skills/abilities and knowledge on general health, enhancing their awareness of healthy behaviors and preventive care. By targeting promotions and campaigns that emphasize the importance of skills and employment, governments can reduce the NEET rates. And, by reducing the NEET rates, governments have the potential to address a broad array of vulnerabilities among youth, ranging from unemployment, early school dropouts, and labor market discouragement, which are all social issues that warrant attention in a growing economy.We also bring to light the health disparities across countries and suggest implications for governments to target educational interventions that can reduce inequalities and improve health, at a macro level. The health effects of education are at the grass roots level - creating better overall self-awareness on personal health and making healthcare more accessible.Our research suffers from a few limitations. For one, the number of countries is limited, and being that the data are primarily drawn from OECD, they pertain to the continent of Europe. We also considered a limited set of variables. A more extensive study can encompass a larger range of variables drawn from heterogeneous sources. With the objective of acquiring a macro perspective on the education\u2013health association, we incorporated some dependent variables that may not traditionally be viewed as pure health parameters. For example, the variable potential years of life lost is affected by premature deaths that may be caused by non-health related factors too. Also there may be some intervening variables in the education\u2013health relationship that need to be considered. Lastly, while our study explores associations and relationships between variables, it does not investigate causality.Both education and health are at the center of individual and population health and well-being. Conceptualizations of both phenomena should go beyond the individual focus to incorporate and consider the social context and structure within which the education\u2013health relationship is embedded. Such an approach calls for a combination of interdisciplinary research, novel conceptual models, and rich data sources. As health differences are widening across the world, there is need for new directions in research and policy on health returns on education and vice versa. In developing interventions and policies, governments would do well to keep in mind the dual role played by education\u2014as a driver of opportunity as well as a reproducer of inequality . ReducinOur study offers several prospects for future research. Future research can incorporate geographic and environmental variables\u2014such as the quality of air level or latitude\u2014for additional analysis. Also, we can incorporate data from other sources to include more countries and more variables, especially non-European ones, so as to increase the breadth of analysis. In terms of methodology, future studies can deploy meta-regression analysis to compare the relationships between health and some macro-level socioeconomic indicators . Future"}
+{"text": "Maxwell\u2019s Demon is a thought experiment devised by J. C. Maxwell in 1867 in order to show that the Second Law of thermodynamics is not universal, since it has a counter-example. Since the Second Law is taken by many to provide an arrow of time, the threat to its universality threatens the account of temporal directionality as well. Various attempts to \u201cexorcise\u201d the Demon, by proving that it is impossible for one reason or another, have been made throughout the years, but none of them were successful. We have shown (in a number of publications) by a general state-space argument that Maxwell\u2019s Demon is compatible with classical mechanics, and that the most recent solutions, based on Landauer\u2019s thesis, are not general. In this paper we demonstrate that Maxwell\u2019s Demon is also compatible with quantum mechanics. We do so by analyzing a particular  experimental setup and proving that it violates the Second Law. Our discussion is in the framework of standard quantum mechanics; we give two separate arguments in the framework of quantum mechanics with and without the projection postulate. We address in our analysis the connection between measurement and erasure interactions and we show how these notions are applicable in the microscopic quantum mechanical structure. We discuss what might be the quantum mechanical counterpart of the classical notion of \u201cmacrostates\u201d, thus explaining why our Quantum Demon setup works not only at the micro level but also at the macro level, properly understood. One implication of our analysis is that the Second Law cannot provide a universal lawlike basis for an account of the arrow of time; this account has to be sought elsewhere. J. C. Maxwell devised his so-called Demon in 1867 to show that the Second Law of thermodynamics cannot be universally true if classical mechanics is universally true. In this paper we show that a Maxwellian Demon is compatible with standard quantum mechanics.Maxwell\u2019s Demon is a way of demonstrating that the laws of mechanics are compatible with microstates and Hamiltonians that lead to an evolution which violates the Second Law of thermodynamics by transferring heat from a cold gas to a hot one without investing work. That is, the Demon is not a practical proposal for constructing a device that would violate the Second Law of thermodynamics, but rather a statement to the effect that the Second Law cannot be universally true if the laws of mechanics are universally true.Attempts to prove the Second Law as a universal theorem in classical statistical mechanics, encounter the following issue: since the fundamental microscopic dynamics is invariant under velocity reversal, one can show that given any Hamiltonian, if there are entropy-increasing trajectory segments in the system\u2019s state space, then there are also entropy-decreasing ones, and so the thermodynamic law cannot be strictly true. Essentially the same issue arises in the context of the standard approaches to quantum statistical mechanics see ,3,4,5,6),4,5,63,4Since Maxwell, many attempts have been made to disprove Maxwell\u2019s intuition. In ,20,21,22The situation just described is in the context of classical mechanics. There is a vast literature  for the thermodynamic notion of entropy According to this line of thinking, the thesis is grounded in the Second Law of thermodynamics . This is the prevalent approach, both in arguments supporting Landauer\u2019s thesis . Importantly, Maxwell\u2019s Demon is not in conflict with the empirical evidence, because the available proofs that Maxwell\u2019s Demon is compatible with fundamental physics leave open the possibility that both Second Law behavior and Maxwellian Demon behavior are compatible with fundamental physics; they may hold for different initial conditions of the universe, for example. Nevertheless, for many thinkers, this last option is not satisfactory, and they strive to prove that Maxwellian Demons are incompatible with fundamental physics. One way of doing so, in fact the most prevalent way in contemporary research, is by relying on Landauer\u2019s thesis , and dynamical evolution that satisfy the laws of mechanics , but violate the Second Law of thermodynamics? Our answer in this paper is in the affirmative.We consider a thought experiment along the lines of Szilard\u2019s (see ) and BenOur discussion is in the framework of standard quantum mechanics in von Neumann\u2019s 1932 formulation (see ). We givThe structure of the paper is this. In Consider the setup in see our ,18,19).,19.t0 a We will now show that according to quantum mechanics this setup can be a Maxwellian Demon. We start by constructing a quantum mechanical microscopic dynamical evolution that satisfies the Schr\u00f6dinger equation at all times . We consider the implications of our setup in standard quantum mechanics with the projection postulate in the next section. In both cases, our dynamical evolution includes measurement and erasure under idealized assumptions. Again, we set aside questions concerning the interpretation of such theories. In the last section we argue that as a matter of principle from our microscopic setup one can construct a quantum mechanical macroscopic Demonic evolution. We discuss in t0 with the particle or the weight (or the partition).At a of the box, where the amplitudes in n is an even number and M is the mass of the particle. In standard quantum mechanics, such a superposition does not express ignorance about the energy eigenstates of the particle. It is the fine-grained complete energy state of the particle. In this sense one can say that such a superposition is the quantum mechanical analogue of a classical microstate.The initial energy state of the particle z-direction. The two other spin-1 eigenstates, z-direction, correspond to the two possible outcomes of the measurement. Another alternative is to take the device to be a spin \u00bd particle, the pointer states in this case would be the up and down eigenstates of (say) the spin in the z-direction, and the initial ready state x-direction).t0 in isolation from the environment. Alternatively, if there is a certain amount of decoherence, we may assume that the degrees of freedom in the environment are controllable in the subsequent stages of our experiment. Of course, assuming that the partition is massive, the idealization of zero width may also be problematic, and consequently the wavefunction may change when the partition is inserted. What we need is a way of inserting a partition such that it will not change the expectation values of the energy of the particle. In this sense we assume here that the effects of the above issues are negligible and we continue with our idealization. Again, we are not concerned here with the experiment\u2019s feasibility, but rather with its consistency with thermodynamics.At a/2, where for x varies over the range  and for x varies over the range , while the equal amplitudes are a consequence of our idealization that the partition is placed exactly in the center of the box. The expectation value of the energy of the particle is unaltered a/2). Although the expectation value for the particle\u2019s energy does not change in this interaction, the wavefunction of the particle now becomes a superposition of two components, as in (3). We assume that there are no other changes in the quantum state of the particle from t0 up to t1.We therefore take it that ideally at At The measurement at time  see our . For an  see our ).At w(t) and u(t), respectively. The expectation value of the particle\u2019s energy t5. The energy state of the weight y(t) ranges from the down to the up position, and the expectation value of the energy of the weight increases accordingly. Conservation of energy, which is stated in terms of expectation values in quantum mechanics according to Ehrenfest\u2019s theorem, implies thatAt Since the weight is positioned in a gravitational field, higher energy expectation values for the weight are coupled with higher positions of the weight. This means that the quantum-mechanical transfer of energy increases the probability that the weight will be found at higher positions upon measurement.t the overall probability of finding the particle in the left- or right-hand side of the box is invariably \u00bd. The conditional probability of finding the particle in the right- (left-) hand side, given that the device is in the state a with amplitudes which match the decrease in the expectation value of the energy, The Hamiltonian is such that during the expansion of the particle, as described by (5), the correlations between the memory states of the device and the energy states of the particle are gradually lost. The energy states e. For this purpose, we let the particle interact with e in such a way that the particle will certainly return to its initial energy state e decreases by the same amount as the expectation value of the energy of the particle (or ultimately the weight) increases, To complete the cycle of operation we need to evolve the particle back to its initial energy state. We carry out this task by extracting energy from the environment  see our , Chaptert6 is this:The quantum state of the setup at the end of the process at Therefore, the cycle is completed and the setup is ready to go once again.What is a completion of an operation cycle? Once the cycle is completed, we do not want the environment and the weight to return to their initial state. Instead, the idea is that at the end of the cycle the situation will be such that the only change in the universe is that energy has been transferred from the environment to the weight as in state (8). Bennett and Szilard, for example, argue that completing the cycle of operation involves dissipation in the environment, and therefore the environment\u2019s final state is a fortiori different from its initial state. For them, not only the state of the environment changes, but the entropy of the environment, increases. However, in our setup there is no reason to think that the transfer of energy from the environment to the weight results in an increase of entropy of the environment. Such increase does not follow from the equations of motion. , the von Neumann entropy does not change, since the environment evolves from one pure state to another). As we said in In our above discussion, we have phrased our argument in terms of entropy. There are different notions of entropy in the literature, for example, thermodynamic entropy (see ); Boltzme) it is eitherLet us now consider our setup in the framework of standard quantum mechanics in von Neumann\u2019s formulation with the projection postulate in measurement. As standardly understood, we take the projection postulate to express the so-called collapse of the quantum state: a pure-to pure (nonlinear and stochastic) state transition of a single setup in measurement. Applying the projection postulate at z-direction) on the device until the outcome corresponding to The unitary Schr\u00f6dinger equation cannot map the final states ann . This shows that a Maxwellian Demon is compatible also with quantum mechanics with the projection postulate.Often in the literature it is argued that the erasure of the memory is accompanied by dissipation, according to the Landauer\u2013Bennett thesis. We have disproved this thesis in the classical case in our ,21,22,57The discussion of Maxwell\u2019s Demon in the literature focuses on the notions of measurement and erasure. In the classical case, because of the determinism of the dynamics, these notions must be understood as referring to a macroscopic evolution, namely evolutions of sets of microstates  (13) does not actually erase the outcome of the measurement, since the Schr\u00f6dinger evolution is reversible. However, this impression is mistaken for the following reason. If one posits that the third arrow in evolution (13) does not bring about an erasure just because the dynamics is reversible, then by the same argument the determinism of the dynamics entails that the first arrow in (13) does not bring about a measurement with a definite outcome. Any account in which one obtains a determinate outcome at the end of the measurement, would also be an account in which the outcome is erased at the end of the erasure.In standard quantum mechanics with the projection postulate in measurement, there are two possible evolutions of the quantum state of the measuring device (since the outcome of the measurement is stochastic), each with probability \u00bd, as follows:As before, the first arrow on the left-hand side represents the measurement, the second arrow represents the erasure and the third represents the return to the ready state . As we saw in In standard quantum mechanics (with and without the projection postulate), as we saw, the measurement of any observable that does not commute with the memory observable is an erasure, since after such a measurement it is impossible to retrodict the pre-erasure state of the memory system from the post-erasure state. In a collapse theory after a quantum erasure, one cannot even retrodict which observables had definite values in the past. In our setup, since  describes measurement and erasure per se, turns out to be immaterial to the issue at stake: what is important here is whether or not our cyclic evolution results in total entropy increase anywhere in the universe. As we saw, we have a reversible quantum evolution of a closed system (including the environment), in which energy is transferred from the environment to the weight; the weight ends up in its up state; at the end of each cycle all other subsystems return to their initial states or to states of equal entropy; and there are no further changes in entropy in the universe. This is why such an evolution is a Maxwellian Demon.In this paper we have shown that an entropy decreasing evolution, which includes measurement and erasure, is compatible with quantum mechanics. The entropy decrease in our setup, which is of a closed system , violates the Second Law of thermodynamics. In the classical case there are no microscopic measurements and erasures. Therefore, the question of Maxwell\u2019s Demon must be analyzed by introducing macrostates and macroscopic evolutions. In standard quantum mechanics, however, measurement and erasure can be constructed microscopically, as we have demonstrated above. In other words, in quantum mechanics there is no need to introduce quantum mechanical macrostates in order to consider the question of Maxwell\u2019s Demon.However, if one is interested in a macroscopic description of a Demonic evolution in quantum mechanics, it is important to note that the classical notions of macrostates and macroscopic preparations do not have trivial counterparts in quantum mechanics nian see  below. TThis result implies that the Second Law of thermodynamics is not a universal theorem of quantum statistical mechanics. Our previous result of a classical Maxwellian Demon  that prevents such re-interference. As we said, we are not interested here in practical limitations. Therefore, environmental decoherence does not impose a quantum mechanical lawlike limitation on our thought experiment."}
+{"text": "Diabetes care often requires collaboration between general practitioners, allied health professionals, nurses, and/or medical specialists. This study aimed to describe the establishment of an integrated diabetes prevention and care approach in an area with limited access to primary and secondary care, and the challenges faced in its initial development.A qualitative research approach to identify challenges was taken. Data included meeting minutes, observational data and reports involving local clinical and non-clinical stakeholders from June 2016- December 2018 and were thematically analysed.Key challenges were low patient attendance in general practice, healthcare professional time, low participation at health promotion activities/peer support groups and diabetes education reflecting a low priority among people with and at risk of diabetes. Coordination between services remained a challenge.This study highlights the need to integrate new diabetes services with existing health activities in the community and the importance of allowing flexibility and regular contact with local healthcare professional and community to encourage their involvement. Regular meetings with the funders, internal and external stakeholders are key for sustainability and to adapt programmes to the local situation. Further work is needed to identify and implement strategies to overcome these challenges. Diabetes mellitus is a significant and fast growing health problem in Australia and around the world . DiabeteA review of integrated diabetes care across the world  concludeWollondilly is an inner-regional local government area  in South Western Sydney, in New South Wales, Australia. The population in 2016 was 48,519, among whom 1,552 were Aboriginal origin dispersed across the district 8. WollondBeing overweight or obese can increase the risk of a person developing type 2 diabetes . ObesityThe aim of this paper is to describe the establishment of an integrated diabetes prevention and care approach, the Wollondilly Diabetes Programme (WDP), and to report on barriers faced in its initial development.Ethical approval was granted by Western Sydney University Human Research Ethics Committee .The Wollondilly Health Alliance was established in 2014 to facilitate joint working between the Local Health District (state funding), the Primary Health Network  and Wollondilly Shire council, to improve health care in the region [The Wollondilly Health Alliance includes a Care process  and a Health promotion (community) working group. Core membership of the Care process working group includes representatives from general practice, non-government organisations and private industry. Core membership of the health promotion working group includes health promotion officers, project managers/officers and community support workers . The WDPLinkage with other Wollondilly healthcare professionals (besides through example practice based work) occurred through new bi-monthly Clinician Reference Group (CRG) meetings. The focus of these meetings was to share information about the progress of all aspects of the programme  and to receive feedback and guidance on how the programme should proceed, provide diabetes healthcare professional education (including case based discussions) and to nurture an integrated approach to diabetes care.The WDP clinical management services are summarised in Figure Case conferencing allows an endocrinologist to visit general practices to advise general practitioners with managing more complex diabetes patients/cases. The case based discussion is structured for both healthcare professional education and clinical decision-making, and builds a relationship between the specialist service and primary care. . Case coHealthPathways is an online referral guidelines portal used by health professionals at the point of care to plan patient care, and where necessary, provide information on how to refer to specialists and other services . The WDPThe Australianised Cambridge Diabetes Education Programme (AusCDEP) is a competency based online learning tool for diabetes related topics that supports all levels of healthcare practitioners. For example, it included basic topics (what is diabetes) for non-clinical administrative staff, such as the receptionist, key issues (e.g. hypoglycaemia) for all clinical staff to advanced topics  for more specialist staff to address the needs and deliver quality care . The WDPThe aim of this component was to encourage Wollondilly residents to take advantage of new and existing health promotion activities running in Wollondilly. There were several activities promoting healthy eating, lifestyle and social support in Wollondilly initiated by the Wollondilly Health Alliance. The team gained information on all health promotion activities, by attending the bi-monthly health promotion working group meetings. The WDP also initiated diabetes awareness approaches including a roadshow, door to door survey and promotion through social media . Figure In a peer support programme, one person supports another with similar personal circumstances. Self-management of diabetes can be supported by peer support facilitators who provide guidance and support on \u2018how to do\u2019 rather than the \u2018what to do\u2019 . Four keNot all residents attended general practices within Wollondilly and therefore nine recruitment approaches were used to recruit participants, described below in Table This qualitative study was conducted in Wollondilly Shire to identify the challenges faced during the programme. It is a prospective observational study using an ethnographic methodology. Data were captured and collated from Table  \u2013 ClinicStudy participants included both internal  and external stakeholders  who attended meetings.Data were thematically analysed by identifying patterns (themes) , managedThis first phase included becoming accustomed with data. Data were integrated and were closely read multiple times before generating codes by one author (RZ). Data were discussed with co-author (FM) by sharing each other\u2019s perceptions to gain better understanding.Initial codes were generated by systematically reading and rereading the data to become more engaged with the data. Codes in the narrative of data were labelled on minutes/notes, resulting in a variety of codes. Coding was cross-checked by a researcher (FM) with extensive experience in qualitative research, to assure accuracy and consistency.Similar codes were organised together and potential themes were explored and built. Extracts related to the codes were reread to explore if they fit better elsewhere or into the existing theme to prevent any contradictions and continue distinction between themes, expressing a story about each aspect of the data clustered. All relevant data were explored to represent data precisely.This phase included final reviewing of themes to identify any linkage. Each theme with its coded and clustered data were reviewed to avoid conceptual overlap. Themes were agreed upon and were identified which reflected participant\u2019s perspectives on the challenges to implementing the WDP.This final phase involved revising the theme names, coding, and all over dataset before final analysis and writing up findings. Final finding are explained in the results section of this paper.A 15-point checklist was used to ensure trustworthiness of data and to confirm that all six phases were followed (Supplementary table S1) .Participants of the observational study included endocrinologists (n = 2), dietitians (n = 2), a credentialed diabetes educator (n = 1), health promotion professionals (n = 5), general practitioners (n = 8), practice nurses (n = 4), pharmacists (n = 5), and other allied health professionals (n = 4) . Three major themes with were identified from the data collected.All the general practices  in Wollondilly were approached to work with the WDP clinical services. Two practices considered themselves self-sufficient about diabetes care and declined invitations to participate in clinical care but attended meetings. In the larger practices in Wollondilly, practice nurses were responsible for identifying suitable patients for consultations. Occasionally they were unable to prepare patients for the next month due to their time commitments. This preparation work was often completed outside of their scheduled working hours, which affected the progress of the programme and the patients who were in need of consultation.Case conferencing was sometimes challenged by the need for a third healthcare professional to attend and variable attendance by practices when patient load was high. Some practices were open only 1\u20132 days per week and therefore had little time for case conferencing. Availability of the endocrinologist was also an issue, with less than one funded day per week. The duration of case conferencing was also an issue as all session times must match and were to be negotiated at the time of the phone call to the practices.The dietitian and the credentialed diabetes educator worked part-time (two days a week) and not throughout the week, which affected the patient appointment bookings due to their limited availability. Most of the patients preferred consultation or group sessions on weekends or late evenings over weekdays, as they travel to work outside of Wollondilly during weekdays. Even after the services were delivered within the practice, with invitations sent from the practice, patient attendance was low which could be due to low patient priority for diabetes education. Smaller practices were unable to physically accommodate group education sessions. The Medicare funded Chronic Disease Management item  limits rThe peer support programme and community engagement activities/events were slower to set up than clinical services. It was a challenge to engage with residents of all the towns due to their geographical dispersal . Most of the events and peer support activities were initially held in two major towns of Wollondilly and only a few residents of other towns and villages would attend due to diabetes education being a low priority and the distance and travel time, with residents having to travel for at least 15\u201335 minutes and for some for up to an hour. It was challenging to conduct surveys and to interact with residents in certain towns of Wollondilly as the properties are large and gated, which made communication with these residents difficult. The WDP was promoted through existing community networks within Wollondilly but their available time and detailed knowledge for promotion were limited.We experienced similar challenges to reach the population living in semi-rural or rural areas as noticed in other studies, such as low attendance and low priority for diabetes education 2728. The27Data collected such as the WDP services uptake, awareness of the WDP through door-to-door survey, number of people joining peer support group was used as a quality improvement tool and a clinical audit of the programme is ongoing to find the impact of the programme on population. The numbers were reviewed every week during the team meetings Table .This study describes in detail the key challenges and lessons learned while implementing an integrated diabetes care and prevention service de novo into an inner-regional area. Most of the following lessons learned may help future projects to increase their integrated care programme/project efficacy and effectiveness.Creating the relationship with primary care was of key importance with the need to overcome the initial hump of work to generate much-needed referrals to specialist endocrinologist, allied health clinics and for case conferencing. Some practices open only 1\u20132 days per week and therefore had little time for case conferencing. We approached all practices and encouraged them to participate. Seven practices (7/11) participated in at least one aspect of the programme. The limit of 5 Chronic Disease Management Medicare items resulted in some allied health/practices seeing the WDP as a competitor. Therefore, podiatry treatment never started and screening was no longer provided to avoid any conflicts. General practices expressed that the systematic referral processes were time consuming to implement, as the process required identifying patients systematically. Time constraints resulted in fewer referrals than expected in spite of the clinical burden  It has been known for over 20 years that diabetes care and patient satisfaction can be improved, by providing support to primary healthcare teams  and the Joint specialist and general practitioner case conference evaluations in Western Sydney and Hunter and New England have been associated with improvements in clinical parameters such as Haemoglobin A1c, Body Mass Index and blood pressure of patients. They have high program acceptability from both general practitioners and patients 31. Such Change management made implementation sustainable when services were delivered in practices avoiding competition. The initial plan was to provide podiatry services, however this was terminated to avoid conflicts with the local allied health. Group education sessions were moved to practices to build rapport with the practice staff and overcome such perception. The WDP dietitian and credentialed diabetes educator contacted patients with type 2 diabetes at the Picton and Wilton practices for the group education. This was useful, and increased numbers of attendance, as often due to staffing issues the individual practice did not have time to call patients. The idea to move the education to individual practices was also to increase trust with patients as it was within their GP practice and to combat the lack of transport for some. Interventions at health service provider level also has shown to have a positive impact on the overall health and lifestyle of patients with type 2 diabetes living in a rural area  including an increase in satisfaction level of both patient and the provider .There were changes to the WDP team in the second year (dietitian and administration staff); however, new roles improved the productivity of certain aspects of the programme (such as community coach for peer support programme), volunteer involvement improved the health promotion aspect of the programme, increasing the community participant and awareness on the WDP. Leadership plays a significant role for successful implementation to address population needs, setting clear goals for the team, managing changes and adding innovative designs to an integrated programme 35. The cA needs assessment consultation process conducted by the Wollondilly Health Alliance reported the community, general practitioner, allied health provider and non-government organisation perceptions of the health services in Wollondilly. Challenges in accessing health services were identified by nearly 60% of survey respondents because of the shortage of general practitioners, resulting in long wait times to see a general practitioner across Wollondilly .Challenges with community participation included slower set up of prevention activities and low attendance sometimes reportedly due to distance and travel time but also a lack of prioritising diabetes education by patients. Community activities/events were initially mostly held in the two largest towns (Picton and Tahmoor) of Wollondilly and residents from other towns had to travel long distances (15\u201335 minutes) to these towns and health service centres, worsened by poor public transport in Wollondilly. Limited public transport was also noted as one of the challenges to a diabetes prevention programme (Greater Green Triangle Diabetes Prevention Project) in a rural area of south eastern Australia . PromotiIt was not possible to have a joint meeting with all working groups at one time mainly due to time restrictions and other commitments. Separate meetings were held with community workers and council. The need for joint working between organisations, inter-professional collaboration, patient awareness and adequate and long-term financial support required a major shift in the \u2018natural\u2019 modus operandus. Staffing issues limited the practice opening times and the inclusion of a third health care professional for case conferencing.Case conferencing was initially a part of the WDP however; after one year, the contract for this service shifted from the Wollondilly Health Alliance to the district-wide contract with the local Primary Health Network. A different specialist was extended to work in Wollondilly who was linked with the hospital clinic in Campbelltown (adjoining suburb/area), but not linked to the clinical aspects of the WDP. As a result, the WDP allied health professionals could no longer readily link to case conferencing. This led to a reduction in the number of patients seen in the WDP clinics. The change in funding came from decisions by the Local Health District; at a district wide committee level not through the Wollondilly Health Alliance committees. The funding arrangement and responsibilities within the system are complex, as they are spilt between all levels of government. In Australia, most hospital care is provided through state government systems, but ambulatory care (both primary and secondary) is funded by the federal government through Medicare (a partly to fully publicly funded fee for service system) . As a reWe could identify and address some of the challenges from the beginning of the programme because of the regular team meetings. Shared clinical priorities improved integrated care delivery through working group meetings and incentives encouraged participation of general practitioners such as for case conferencing. Continuous professional development platform was offered through Clinicians reference group meetings to support joint working, and innovation for better care delivery. Data were used as a quality improvement tool to help identify and address any gaps. Regular meetings with both internal and external stakeholders were key in addressing the challenges and increased community participation.We collected data based on human experience and included the feedback/comments from clinical and non-clinical staff who participated in the programme. The challenges faced have been examined throughout the duration of the delivery of the programme, so that adjustments/solutions can be identified promptly to address these. Meeting minutes were collected from the start of the programme, which allowed us to understand the barriers from programme initiation. A second researcher independently verified qualitative data coding to ensure the trustworthiness of this analysis. A major strength of the study is the transferability of the lessons regarding the challenges, and the key interventions .A limitation of this study is that it only includes the views of health professionals involved in the programme and lacks patient feedback regarding their view on diabetes care delivered in their area. Interviews with patients and other stakeholders are planned, which should inform further enhancements to the programme. There was inconsistency in attendance at meetings and some attendees might not have provided in-depth information on highlighted issues, affecting the interpretation of the collected data. The study only covered an inner regional area and hence, does not have national generalisability. A further limitation is that only the processes and challenges have been presented. Outcome studies are underway, separately evaluating the clinical, case conferencing and health promotion components.This study has described the implementation of the WDP along with the challenges faced at various stages and found that to provide seamless integrated diabetes care, both external and internal stakeholders need to work closely, but coordination and joint working requires additional time and supportive systems. Significant challenges occurred, in spite of substantial need, and the alignment of the WDP with the goals of \u2018The Australian National Diabetes Strategy 2016\u20132020\u2019 including improving the co-ordination of resources by all levels of government . Major hWhile the WDP aimed to provide better access to diabetes care by inter-professional and inter-organisational collaboration, a fuller, quantitative evaluation is required to assess the extent to which care was improved, and its associated costs. Further work is also needed to gather in depth perspectives on the challenges identified, to help find strategies to overcome these barriers. This implementation approach can be applied to similar geographical regions and population spread in other countries. Considering the lessons learned from this integrated care case could guide the development of an efficient approach in a similar setting.The additional file for this article can be found as follows:10.5334/ijic.4692.s1Supplementary table S1.A 15-point checklist of criteria for good thematic analysis."}
+{"text": "Accurate target detection and association are vital for the development of reliable target tracking, especially for cell tracking based on microscopy images due to the similarity of cells. We propose a deep reinforcement learning method to associate the detected targets between frames. According to the dynamic model of each target, the cost matrix is produced by conjointly considering various features of targets and then used as the input of a neural network. The proposed neural network is trained using reinforcement learning to predict a distribution over the association solution. Furthermore, we design a residual convolutional neural network that results in more efficient learning. We validate our method on two applications: the multiple target tracking simulation and the ISBI cell tracking. The results demonstrate that our approach based on reinforcement learning techniques could effectively track targets following different motion patterns and show competitive results. Tracking individual cells in a group is the fundamental of many biomedical analysis tasks, including understanding how genotypes are related to phenotypes, tracking the early development of organs and meristems, and potentially tracking the development of cancerous tumors  to extract features from a set of training samples and use the pointing mechanism  A simple framework for cell detection and association based on the idea of (2) We introduce a formulation that translates the decision making in the linear assignment problem algorithm into an RL problem. (3) We propose a novel neural network architecture that end-to-end maps the inputs to the decision outputs.The organization of this paper is as follows. Related work is introduced in section 2. The framework of the proposed method and training details are presented in section 3. In section 4, some experiments are conducted to evaluate the performance of our proposed method. The conclusion is given in section 5.A large variety of cell tracking methods have been described in the existing literature. These cell tracking methods can be broadly grouped into two categories: (i) tracking by model evolution and (ii) tracking by detection.In tracking by model evolution methods, cell segmentation and tracking are solved simultaneously in each frame of a cell video. Typically, these methods are driven by data in some feature space and make a regularity assumption on the smoothness of the curve. In this framework, cells are represented by parametric or implicit active contour models. Parametric models utilize the explicit representations of cell boundaries such as Gaussian Mixture Models (GMM)  Problem Formulation. We formulate the procedure for selecting an assignment solution as an RL problem to associate target states and measurements. (2) Neural network architecture. An end-to-end architecture that maps from the state space to the action space is designed. (3) Training algorithm. We present the RL algorithm used for the policy search.t \u2212 1. Each element of \u03a9t\u22121 corresponds to a cell trajectory. To find their associated new measurements at time step t, each trajectory would be predicted by a Kalman filter and then find the possible association between predicted cell states and new measurements. Let the set t \u2212 1. Then the association mapping from set B to the measurement set Assume that the cell trajectories can be denoted as a set D are calculated through the location distance between the elements of set B and the measurements as shown in D is defined asThe values of the cost matrix D is a (Mt\u22121 + Nt) \u00d7 (Mt\u22121 + Nt) square matrix, with the row and column indices representing the Mt\u22121 prediction from trajectories and Nt measurements. The matrix D consists of four sub-matrices \u039b(Mt\u22121 \u00d7 Nt), \u03a5(Mt\u22121 \u00d7 Mt\u22121), and \u0393(Nt \u00d7 Nt) implies that the corresponding target's state is judged as \"Tracked\", \"Lost,\" and \"New,\" respectively. In the sub-matrices \u03a5(Mt\u22121 \u00d7 Mt\u22121) and \u0393(Nt \u00d7 Nt), we define the value of the diagonal element as a distance threshold and other elements to be \u221e. Here, when a predicted state is highly self-associated, we consider it to be lost. An estimated state that highly associates itself is considered as a new target. The elements of the sub-matrix \u039b(Mt\u22121 \u00d7 Nt) are the distances between the prediction state and measurements.where t \u2265 0, an agent is in a state rt \u2208 \u211d and transitions to the next state st+1 ~ p. A policy st). The objective of RL is to search for a policy that maximizes the expected cumulative rewards over a horizon T, i.e., p]. In practice, we consider parameterized policies \u03c0\u03b8 and aim to find The standard RL formulation starts with an MDP: at time step rt and transition dynamics st+1 ~ p.To formulate the procedure of selecting assignment solution algorithms into an MDP, we specify below the state space State SpaceAction SpaceRewardrt. For most RL applications, designing a reward function is always a critical part, especially when the agent needs to precisely perform actions in a complicated task. A good reward function will make the agent learn more efficiently and achieve better results. By contrast, an agent with a poor reward function may suffer slow convergence or even produce undesirable results. The objective of the linear assignment problem is to minimize the total cost of the assignment solution. To achieve this objective, we design the reward function as the sum of the assignment cost after producing an assignment solution. Given a cost matrix C = {cij}, i = 1, \u2026, N, j = 1, \u2026N and a selected assignment solution X = {xij}, i = 1, \u2026, N, j = 1, \u2026N, the reward rt can be defined as Transitionst. In our work, the state transition is deterministic after an action has been chosen because it can directly assign the corresponding task to the person.C that can be treated as the sum of a probability distribution for matching X and a noise V asThe input of our residual CNN (ResCNN) is a cost matrix The sequence-to-sequence models the linear assignment problem  and a rectified linear unit (ReLU) . Finally, the probability distribution is clipped by the tanh activation function so that the intensities of the output lie in the range .Our proposed neural network is similar to the image denoising network introduced in Zhang et al. . The inpIn summary, the main feature of our ResCNN is the adoption of residual learning to learn g et al. , batch nIn the following, we will give some important details about our network design and training.t before non-linearity as follows:Batch normalization is a standard technique that is widely used in image classification CNN models. Training a deep neural network model is often difficult not only because of the gradient vanishing/exploding problem but also because the distribution of data changes between layers, which is called the \u201cinternal covariate shift\" phenomenon. Batch normalization is a technique that can relieve this phenomenon by introducing several simple operations to the input data. The goal of the normalization step for batch normalization is to transform the layer input t] and Var[t] are the expectation and variance computed over all training data. It is usually impractical to exactly calculate E[t] and Var[t] with stochastic optimization. Batch normalization instead approximates E[t] and Var[t] via the mini-batch statistics during training. It would be beneficial if the mini-batch statistics agree well with the full training data statistics.where E[Batch normalization and residual learning are two important algorithms for designing a neural network architecture. Residual learning and batch normalization can benefit from each other Unlike ordinary visual tasks, for the linear assignment problem, one major characteristic is that one detected cell can only be assigned to one predicted cell. The neural network output should satisfy one-to-one constraints. Let uij is the probability that predicted cell i at time t \u2212 1 is assigned to detected cell j at time t. i\u2032. Next, a normalized softmax operation is applied to u to compute the final output probability matrix.where C = cij. The output of the network is the assignment solution \u03c0. In this work, we use the sum of the selected costs C defined as follows:In this paper, we utilize the RL to train the neural network. The input of the network can be denoted as p is the stochastic policy of a neural network with parameters \u03b8. We learn \u03b8 using the Adam optimizer based on the REINFORCE algorithm  denotes the baseline value of the assignment cost and is used to reduce the variance of the gradients. If we randomly obtain Mi.i.d. samples, then the above gradients can be approximated bywhere b(Ci) is initialized by calculating the sum of the cost of the assignment solution that is generated by the neural network. In each step, the baseline value is updated as follows:For a cost matrix, the baseline value Algorithm 1 gives the pseudo-code of the training procedure of the neural network.To evaluate the performance of our proposed method, we consider two applications of the linear assignment problem: maximum weight matching (MWM) and data association for multi-target tracking. We first compare our method with the state-of-the-art DRL method for maximum weight matching. Then, we test our method on a multi-target tracking scenario. Finally, we evaluate our proposed method on three cell microscopy datasets, Fluo-N2DH-GOWT1, PhC-C2DH-U373, and Fluo-N2DH-SIM+ from the ISBI 2015 Cell Tracking Challenge , where V is the vertex set containing two disjoint vertex sets V1 and V2, with |V1| = N and |V2| = N, and E is the set of all edges between every node v1 \u2208 V1 and v2 \u2208 V2. Let wij, i \u2264 i \u2264 N, 1 \u2264 j \u2264 N denote the associated weight for the edges in the graph. Then, a matching in a graph G is a subset of E such that no two edges share a common vertex. A maximum weight matching is a matching such that the sum of the weights of the edges in the matching is maximal , and Wij is the Euclidean distance between vertex i and j. We select the optimality ratio as Define a weighted bipartite graph N = {15, 20, 25}. The results are compared with SPG+Matching and AC+Matching  and velocity  information. zt = Hxt + vt, where R = 0.05I2 \u00d7 2.One major application of linear assignment is data association for multi-target tracking. Therefore, we set up a simulated multi-target tracking scenario to evaluate the performance of the proposed method similar to Milan et al. . Five taC \u2208 \u211dN\u00d7N is the Mahalanobis distance between the estimated target states and the measurements. We compare JPDA-RL with the traditional joint probabilistic data association (JPDA) filter  and Number of Identity Switch (IDSW). The OSPA-T distance  is the L2-norm, \u03a0n is the permutations in {1, \u2026, n} and where c and outlier sensitivity p, need to be set. In our simulations, we set c = 1 and p = 1.To compute the OSPA-T distance for the estimated tracks and true tracks, two parameters, the cardinality penalty In The segmentation task by U-Net and data association by DRL are conducted on AMD Ryzen 9 3900X 12 core processors with a GeForce GTX 2060 graphics card. For comparison, segmentation (SEG), tracking (TRA) accuracy measures and overall performance (OP) are adopted to evaluate the tracking performance. For TRA, Acyclic Oriented Graph Matching (AOGM) is used to count the changes needed to transform the cell tracking family tree into the ground-truth graph. OP is defined as the mean of TRA and SEG.Fluo-N2DH-GOWT1-01 dataset, we compare our method with the two tracking-by-detection [CPN   (Gupta et al., The results of this work are compared against the best performing available methods for each dataset. For the Fluo-N2DH-GOWT1-01 sequence. However, our method has a lower TRA score over the Fluo-N2DH-GOWT1-02 sequence. One reason for the lower TRA score of our method is that the Fluo-N2DH-GOWT1-02 sequence has multiple cell events, including mitosis, apoptosis and cell fusion. Our method does not consider the complex process of cell differentiation.PhC-C2DH-U373 sequences, the U-Net tracking method uses the cell segmentation model trained from two sequences. Therefore, the SEG score of U-Net is the best among all algorithms over the PhC-C2DH-U373 sequences. However, even with that advantage, our method still obtains a higher TRA score on the PhC-C2DH-U373-01 sequence. U-Net produces very accurate cell segmentation masks on PhC-C2DH-U373 sequences, but for the data association step, it often fails to associate correctly. The reason is that U-Net utilizes the greedy search method to link the cell segmentation between frames.For the Fluo-N2DH-SIM+ sequences, our method has similar performance with U-Net-S. Once the cells have been detected, our method for cell tracking is able to achieve high overall accuracy in linking the cells between frames.For In this paper, we presented a solution to the problem of data association in cell tracking using the deep reinforcement learning. We formulated the data association problem into a linear assignment problem and then proposed a deep reinforcement learning framework which utilizes a residual CNN neural network. In simulation results, we compare the proposed method with other state-of-the-art approaches on various cell tracking datasets, and the results show that the proposed method achieves better comprehensive performance. Thus, our method likely has applications in the field of biomedical engineering. There are also some limitations of our tracking method that leave room for improvement. In future research, we plan to improve the data association method to deal with one-to-many and many-to-one association problems.All datasets generated for this study are included in the article/supplementary material.LZ and JW substantially contributed to the conception and design of the study. XS analyzed and interpreted the data. LZ, JW, and JZ drafted the article.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest."}
+{"text": "A higher degree of insomnia symptoms at baseline was associated with improvement after IPRP. In conclusion, IPRP seem to have beneficial effects on insomnia symptoms in chronic pain patients. Nevertheless, almost half of the patients still suffer from clinical insomnia after IPRP. The possible effect of systematic screening and treatment of insomnia for improving the effect of IPRP on pain is an important area for future research.Insomnia and chronic pain are prevalent health complaints. Previous research has shown that they are closely associated, but their interaction and causality are not completely understood. Further research is needed to uncover the extent to which a treatment strategy focusing on one of the conditions affects the other. This study aimed to map the prevalence of insomnia symptoms among patients in interdisciplinary pain rehabilitation program (IPRP) and investigate associations between the degree of insomnia at baseline and the treatment outcome regarding pain intensity, physical function, social function, mental well-being, anxiety, and depression. Of the 8515 patients with chronic pain, aged 15\u201381 who were registered in the Swedish Quality Registry for Pain Rehabilitation during 2016\u20132019 and participated in IPRP, 7261 had follow-up data after treatment. Logistic regression analysis was used to investigate associations. The prevalence of clinical insomnia, according to Insomnia Severity Index (ISI), among chronic pain patients in IPRP was 66%, and insomnia symptoms were associated with both country of birth and educational level. After IPRP, the prevalence of clinical insomnia decreased to 47%. There were statistically significant associations between the degree of insomnia symptoms before IPRP and physical function ( Chronic pain is a widespread health problem throughout the world, and is one of the most common causes of sick leave in Sweden . GloballChronic pain often affects mental health negatively, thus many patients also suffer from psychiatric co-morbidity in terms of anxiety, depression, stress and sleep disorders. The relationship between pain and mental health appears to be bidirectional, via the pain modulatory systems in the central nervous system  psychological factors such as mood, anxiety level, memories, attention and distraction, stress, fatigue, and expectations can either increase or decrease the pain experience .The bidirectional relationship between sleep and pain is of special interest. Poor sleep is common in chronic pain patients and recent data identifies sleep problems as key factors in the patients with severe pain presentations ,7. In a Studies suggests that disturbed sleep is an important modulator and cause of perceived pain ,10. ExamThus, there seem to be a bilateral connection between pain and sleep disturbance, even though the exact mechanism of action still needs to be eluded .Insomnia is a symptom comprising decreased quality or quantity of sleep, despite adequate attempts to sleep or maintain sleep. It affects about 10% of the global population . InsomniThe Insomnia Severity Index (ISI) is a validated self-assessment tool for insomnia symptoms. It consists of seven questions giving a total score of 0\u201328 points that can be categorized as follows: no insomnia (0\u20137 points), sub-threshold insomnia (8\u201314 points), moderate insomnia (15\u201321 points), and severe insomnia (21\u201328 points) . An ISI Chronic pain is a complex medical condition in which many factors interact and affect the quality of life, such as high pain intensity, decreased mental well-being, reduced ability to work, and insomnia . PatientThe focus in this type of pain rehabilitation is not primarily to reduce the pain, but to find a sustainable approach to the pain condition where reduced pain can be a positive side effect. Pain rehabilitation focuses on the patient learning strategies to be able to manage the pain and its consequences, increasing the patient\u2019s understanding of the condition so that the patient has better opportunities to cope with daily activities and to be able to return to work.The rehabilitation within IPRP takes place over a period of about three months, and in addition to medical and care-related measures from doctors and nurses includes rehabilitative measures at home and at work, planning and guidance of physical activity by physiotherapists, and mapping and treatment of anxiety, depression, and sleep difficulties by a psychologist.The aim of the present project was to study insomnia symptoms among patients with chronic pain, firstly in terms of the association between the prevalence of insomnia symptoms and sociodemographic factors or pain characteristics, and secondly in terms of the association between the degree of insomnia symptoms at baseline and the treatment result regarding pain intensity, physical function, social function, mental well-being, anxiety, and depression.Data from the Swedish Quality Registry for Pain Rehabilitation for the years 2016\u20132019 were retrospectively analyzed. Data in the registry were gathered via questionnaires filled in by patients on three different occasions: before start of IPRP, immediately after IPRP, and one year after end of IPRP. In the present study, the first two time points are used, before start of IPRP and immediately after. The questionnaires contain several validated self-assessment tools such as a numeric rating scale (NRS) for pain, the ISI , the HosAll patients included in this study were between 15\u201388 years old, with chronic pain treated at one of the 40 pain clinics that report to the Swedish Quality Registry for Pain Rehabilitation. For the period 2016\u20132019, the register contains 23,235 patients who were referred to a pain clinic, 8515 patients who were admitted to IPRP, and 7261 patients who completed the IPRP and answered the questionnaires after the IPRP. The study group at baseline for the present study consists of the patients who were admitted to IPRP, and the study group who completed IPRP consists of those who underwent IPRP and filled out the questionnaires in connection with end of IPRP. The selection process is summarized in The ethics application for access to the registry in order to research multimodal pain rehabilitation was approved on 26 June 2015 (ref: 2015/108\u201331).\u00ae Statistics software package. The ISI was used to assess sleep, with scores divided into four categories of insomnia severity , as described earlier and as used both by the developers of ISI and in research studies using the Swedish translation [Statistical analyses were performed using the IBM SPSSnslation ,27. For nslation . SociodeIn a cross-sectional analysis of baseline data, logistic regression was used to investigate the association between the outcome of clinical insomnia and sociodemographic and pain characteristic factors. The explanatory factors were age, gender, country of birth, education, pain intensity, pain frequency, pain duration, and anatomical distribution of pain (number of pain sites). Logistic regression was also used to investigate associations between the four categories of degree of insomnia at baseline and treatment outcomes regarding pain intensity, mental well-being, physical function, social function, anxiety, and depression reported after the treatment. The treatment outcomes were defined as binary variables (improved vs. worsened), with the minimal clinically important difference (MCID) for each variable being used to calculate an improvement or deterioration. MCIDs used were 2 for pain intensity according to the NRS , 5 for ep < 0.2 were then put together in a multivariable model, and variables with p \u2265 0.2 were removed from the model one at a time according to their p-value.The following procedure was used for both the logistic regression analyses. No collinearity was seen between the explorative variables. This was checked with paired Spearman correlations for all variables, but also with box plots for pairs of a categorical and a continuous variable and pairs of two categorical variables. Separate logistic regression analyses were performed with one explanatory variable at a time, and with insomnia as the dependent variable. The variables that fell out with n = 1814) and 79% were women (n = 6701). The mean age was 45 years  in the total group, 46 years (SD: 12) among the men, and 45 years (SD: 11) among the women. Further background data on the study group at baseline are given in The study group at baseline consisted of 8515 people, of whom 21% were men  and 79% were women (n = 5743). The mean age was 45 years  in the total group, 46 years (SD: 12) among the men, and 45 years (SD: 11) among the women.The study group completed IPRP consisted of 7261 people, of whom 21% were men  as a dependent variable and degree of insomnia before treatment as an independent variable showed some statistically significant results. Odds ratios for improvement in physical function, social function, and mental well-being were all higher when the degree of insomnia before treatment was higher  before they entered an interdisciplinary pain rehabilitation program, and although the prevalence decreased after the program, almost 50% of the patients still reported clinical insomnia. Clinical insomnia was associated with demographic factors, with depression and anxiety, and with increased severity of pain . A high level of pre-treatment insomnia was associated with improvement in functional outcomes, but with non-improvement in mental health.Our finding of a high prevalence of insomnia in people with severe chronic pain is in line with previous studies ,32, as iOur results showed that a higher degree of pre-treatment insomnia was associated with higher odds of improvement in physical function, social function, and mental well-being. One possible explanation may be related to those with a higher degree of pre-treatment insomnia having a higher potential for improvement. This could be investigated in the future by analyzing whether the people who improved in these areas, and had a high degree of pre-treatment insomnia, also improved with regard to insomnia.However, our results showed that a high degree of insomnia before treatment meant lower odds of improvement regarding depression and anxiety. Previous research has shown a strong link between anxiety and depression and insomnia ,37. An eOne of the strengths of this project was the relatively large number of people whose data could be analyzed. The fact that the register from which the data came has good coverage of pain rehabilitation and that it is based on validated self-assessment tools such as NRS, ISI, HADS, and RAND-36 strengthens the validity of our results. Conversely, although the questionnaires used to gather data were based on well-validated self-report instruments, the use of questionnaires and self-reported conditions could be considered a weakness. Since the Swedish Quality Registry for Pain Rehabilitation only includes patient-reported outcome measures, there is no information in the registry regarding diagnoses or treatment. Hence, the impact on insomnia of specific pain diagnoses, co-morbidity and pharmacological therapy cannot be assessed in this study. Another limitation of the study is that only descriptive statistics were calculated regarding differences between men and women. However, gender was adjusted for in the regression analysis.This study shows that the high prevalence of clinical insomnia according to ISI in this patient group persists after IPRP. Nevertheless, both pain intensity and the prevalence of clinical insomnia were lower in the group who had completed IPRP. The findings regarding the connections between insomnia and the sociodemographic and pain characteristics, as well as the connection between the different treatment outcomes and degree of insomnia before treatment, require further research in order to be able to draw additional conclusions. However, the results indicate that chronic pain patients in IPRP with severe pain presentation in terms of high-intensity, persistent and wide-spread pain seem to be a high-risk group with regard to prevalence of clinical insomnia according to the ISI.The results also indicate that chronic pain patients with higher degree of insomnia symptoms seem to improve more with regard to physical and social function as well as to mental well-being. Giving patients this information regarding possible treatment effects before start of an IPRP might further motivate the patients to participate in the IPRP. However, further studies are needed in order to confirm the clinical relevance of these findings as well as effects of insomnia targeted treatment in chronic pain patients undergoing IPRP. These findings raise the question of whether screening and treatment of insomnia could further improve the total effect of IPRP."}
+{"text": "Low- and middle-income countries urgently need to improve emergency medical services (EMSs) as a component of their healthcare systems. Here, we detailed EMS resources and their provision in Hanoi, Vietnam, and discussed necessary policies to upgrade EMSs. Between 2013 and 2018, EMS resources, measured as provider-to-population and ambulance-to-population ratios, decreased, whereas service provision, measured as the number of patients transported by ambulance per population, increased. EMS resources and their provision in Hanoi are far below the standards of high-income countries or figures in neighboring Asian countries. Therefore, it is imperative to upgrade health policies for the appropriate allocation of healthcare resources to EMSs and hospital services. In many low- and middle-income countries (LMICs), the number of patients with various life-threatening conditions is increasing because of the acute onset of noncommunicable diseases and injuries. Despite this, patients frequently face problems such as remote hospitals, insufficient EMS functions, and lack of transportation (2). These changes have resulted in an increase in the number of patients who require emergency care due to conditions such as ischemic heart disease, stroke, and injuries. This is evident in urban areas such as Hanoi, the capital city of Vietnam, which experienced a rapid increase in population, residential areas, and road traffic (the population was 8.05 million in 2019). Nonetheless, Hanoi\u2019s EMSs are unable to meet the increasing need for their services due to their lack of functionality, and most critical patients, including those with out-of-hospital cardiac arrest, have to travel to hospitals by themselves (3).Vietnam has a growing demand for EMSs because of a rapid epidemiological transition in the 1990s: a decrease in the incidence of communicable diseases that used to be dominant causes of mortality (accounting for 52% of deaths in 1986 and 12% in 2018), an increase in the incidence of noncommunicable diseases (from 42% to 63%), and external causes, such as injuries (6% to 24%) during the same period Healthcare communities and policy makers in LMICs urgently need to attend to the issue of unmet demands for EMSs. Thus, on the basis of the data from Hanoi, Vietnam, we detailed insufficient resources and current service provision in EMSs and discussed the type of resources that need to be allocated in such regions.The city of Hanoi controls its EMS. It was established in 1975, following the French model of ambulances staffed with physicians and nurses, and it provides free services to the entire city. The call command center is named the \u201c115 Hanoi Emergency Center,\u201d after the universal three-digit phone number (115) for EMS calls nationwide. It dispatches an ambulance from the nearest of the five dispatch stations in Hanoi. The center and dispatch stations are all independent of hospitals, and the EMS physicians and nurses only provide care in the ambulances and do not work for hospitals.From 2013 to 2018, the number of EMS resources in Hanoi has decreased according to data from the 115 Hanoi Emergency Centre . The num(1) or ratios in some HICs at 0.8-3.2 per 100,000 people(4), even lower than that of neighboring LMICs in the early 2010s  (4). Similarly, the provider-to-population ratio was lower than that of HICs at 4-56 per 100,000 people (4). These shortcomings worsened over recent years because the number of resources decreased while the population increased, possibly owing to insufficient budget allocation for vehicle maintenance and procurement and for staff recruitment and salary, despite the presence of adequate resources in tertiary care hospitals.The EMS resources in Hanoi do not meet the population\u2019s needs. The ambulance-to-population ratio  was much lower than the standards for urban areas in high-income countries (HICs), i.e., 2-3.3 per 100,000 people Moreover, the recruitment of new EMS personnel is facing several challenges. First, although physicians and nurses should undergo an 18-month clinical training program in inpatient settings after graduation to acquire their complete clinical license, EMSs are not considered inpatient facilities, which makes obtaining post-graduate training difficult. Second, their working conditions include environments such as highway accidents and violent scenes, exposing them to the risk of trauma.(4) or the standard response times in HICs  (5), if not extremely long compared to neighboring LMICs  (6). The delay in response primarily results from traffic congestion and drivers not making way for ambulances. Owing to this delay, people would rather use a taxi or private vehicle than wait for the EMS even when transporting critical patients, which would affect patient outcomes. A recent multicenter study in Hanoi showed that only 21% of cardiac arrest patients transported to tertiary care hospitals used EMSs (3).The EMS response time of approximately 15 min in Hanoi is much longer than that of HICs (5-10 min) (7). Despite regional variations in population structure and epidemiological situations, we can assume that similar needs for EMSs exist in Hanoi. Thus, the small number of transported patients implies that most of these needs remain unmet. Many LMICs share the same issue of low utilization of EMS.Although service provision in EMSs is rapidly improving, the number of transported patients  is lower than that in HICs  or neighboring LMICs  Cooperation and communication between EMSs and emergency departments of tertiary hospitals are currently not sufficient. As a rule, the EMS personnel are supposed to select the nearest hospital suitable for the patients\u2019 conditions and contact the emergency medical team of the selected hospital in advance so that they can prepare for the treatments. In reality, however, most transfers by EMS in Hanoi are made without prior notice.Other areas of Vietnam have similar or even worse EMS situations, except Ho Chi Minh City, the largest city in the country, which has a control center and 34 dispatch stations. Among 63 provinces and large cities, only 15 have EMSs (115 emergency centers) with limited functions. Additionally, most of the 115 centers have outsourced their dispatch services to private entities, which means that untrained personnel are managing ambulance dispatch, such as taxi allocation.People are reluctant or unable to use EMSs because of various barriers. An insufficient number of EMS units result in a longer response time that is aggravated by traffic congestion. Many people do not know how to use EMSs and lack awareness on the role of EMSs. Even if they are aware, people may not use EMSs because of their low quality of care. These barriers are particularly severe in remote areas. For example, patients must pay the cost of long-distance travel even when using an EMS, and reaching the hospital can take hours or even days. These barriers widen the gap between those who do and do not have good access to healthcare services.In contrast to the scarce resources for EMS, hospital resources greatly increased in Hanoi during this period. The Ministry of Health and the City Government of Hanoi implemented a plan to upgrade and expand the existing 20 tertiary care hospitals and build 15 new hospitals with a total of 5000 beds by the year 2020. This plan includes the securing and training of many medical personnel to perform hospital services, which would cause an outflow of human resources from EMS.Policymakers need to recognize the importance of EMSs as a component of the healthcare system and provide balanced resources to in-hospital and pre-hospital care components. EMSs contribute to improving access to emergency care as the first point of contact to healthcare by providing rapid transport to the hospital and providing first aid to patients at the scene. Quality hospital care can only be provided appropriately when people have good and equitable access to the care.(6). Additionally, improving public relations to raise people\u2019s awareness of the roles of EMSs is crucial in garnering their support for these policies, appropriate EMS use, and facilitating EMS activities.More resources should be allocated from hospital improvement projects to EMS development. For example, maintenance and procurement of equipment, facilities, and vehicles require a significant budget. To strengthen human resources, the disadvantages of EMS personnel can be minimized as follows: modifying the licensing system, improving staff training and supervision, increasing their salaries, and enhancing safety. When resources are insufficient to develop EMSs based on expensive HIC models, such as the French physician-based system, low-cost and basic, yet effective models should be adopted to increase EMS coverage. Paramedic-, medical assistant-, or volunteer-based systems implemented in other LMICs such as Thailand and Malaysia may also be useful in Vietnam In conclusion, insufficient resources have resulted in large, unmet EMS needs in Hanoi. Policymakers should recognize the importance of EMS as a healthcare component and allocate additional balanced resources. They need to address the challenges faced by EMS personnel to ensure that EMS resources and provision indicators in Hanoi meet the standards of those in HICs.NoneThis work was supported by the Japan Society for the Promotion of Science KAKENHI (16K11422 and 19K09403); the funding body did not play any role in conducting the study or preparing the manuscript.BHH and SN conceived the idea; BHH, THM, TSD, TN, and TAD collected information, analyzed the data, and drafted the manuscript; and BHH, VCL, TN, QCL, and SN critically revised and rewrote the manuscript.The data used in this study are public domain data available from the Hanoi dispatch center.Because this study used public domain aggregated data, requirements for ethical clearance and informed consent were waived."}
+{"text": "Animal derived waste, if not disposed properly, could pose a threat to the environment and its inhabitants. Recent advancements in biotechnological and biomedical interventions have enabled us to bioengineer these valuable waste substrates into biomaterials with diversified applications. Rearing and processing of poultry, cattle, sheep, goat, pig, and slaughterhouse waste can aid in effective waste valorization for the fabrication of biopolymers, composites, heart valves, collagen, scaffolds, pigments and lipids, among other industrially important biomaterials. Feathers and eggshell waste from the poultry industry can be used for producing keratinous proteins and biocomposites, respectively. Cattle dung, hoofs and cattle hide can be used for producing hydroxyapatite for developing scaffolds and drug delivery systems. Porcine derived collagen can be used for developing skin grafts, while porcine urinary bladder has antiangiogenic, neurotrophic, tumor-suppressive and wound healing properties. Sheep teeth can be used for the production of low-cost hydroxyapatite while goat tissue is still underutilized and requires more in-depth investigation. However, hydrolyzed tannery fleshings show promising potential for antioxidant rich animal feed production. In this review, the recent developments in the production and application of biomaterials from animal waste have been critically analyzed. Standardized protocols for biomaterial synthesis on a pilot scale, and government policy framework for establishing an animal waste supply chain for end users seem to be lacking and require urgent attention. Moreover, circular bioeconomy concepts for animal derived biomaterial production need to be developed for creating a sustainable system.  This laThe traditional methods of waste management deal with composting, vermicomposting, biogas production, and value-added product formation that have been extensively investigated ,3. HowevTo our knowledge, there is no available review that provides an account of the biomaterials that can be produced from different categories of animal waste. Therefore, this review focuses on the production of various biomaterials with animal waste as initial substrates for various biomedical applications.2.Globally, poultry slaughter industry introduces 7 billion tonnes of feather waste annually to the solid waste biomass. It was reported that the feathers contained >90% keratinous protein and may serve as a low cost and abundant protein source . Various3 of the composite in comparison to pure epoxy. Furthermore, the increase in the concentration of chicken feather fiber from 0% to 5% reduced 3.79% of the weight density i.e. from 1132.08 kg/m3 to 1089.12 kg/m3, and a further rise in feather concentration to 7% decreased the density to 1078.51 kg/m3 [2 whereas upon addition of 7% of chicken feather fiber, the impact strength increased to 1480\u00a0J/m2. Unlike this, the tensile strength decreased with increasing concentration of chicken feather fiber and the reason for this was the irregular shape and the poor strength of the fibers [Recently, Oluba et al. fabricated a biocomposite film by utilizing ginger starch in combination with chicken feather derived keratin . The ince fibers .3), it can potentially serve as a raw material for the synthesis of biomaterials. The solution of CaCO3 when mixed with H3PO4 and NaHCO3 yields carbonated-hydroxyapatite (CHA) in the presence of microwaves. The CHA is also the primary inorganic constituent of bones in humans. Thus, this can be efficiently utilized for the fabrication of scaffolds to be used in bone tissue engineering [Another major waste generated from poultry is the shell of chicken egg. Since chicken egg shell contains 94% of calcium carbonate (CaCOineering . It was ineering . The morineering . Differeineering . In a stineering . The addineering . Furtherineering .in vivo assay on Wistar rats (Rattus norvegicus), it was recorded that the highest lymphocyte cell proliferation was found in the bone defects treated with chitosan-collagen scaffolds with 50:50 gel mixing as compared to individual collagen and Natrium-Carboxy Methyl Cellulose (3%) [The organic waste released from poultry industry is rich in collagen. Gronlien et al. Isolated collagen from Turkey was soluble in pepsin . The isoose (3%) .3.Staphylococcus aureus (MRSA), non-MRSA and Escherichia coli [2) powder and 95.7\u00a0wt.% natural hydroxyapatite by employing a mechano-chemical method. The synthesized fluorine-hydroxyapatite exhibiting a sphere distribution and had a 80\u201390\u00a0nm crystal size [Among the waste generated by cattle, the excreta and the bones were most studied for their utilization in the fabrication of different products for economic and social value . The biohia coli . Furthertal size . To imprJellification process rapidly solidifies the hydroxyapatite starch composite. The addition of starch to hydroxyapatite guarantees the bimodal size distribution that leads to a decrease in the porosity and improved strength of the jellified product . The hyd\u22121 and 0.45 to 0.55\u00a0\u00b5m\u22121, respectively. This small change in the volume density and surface to volume density suggested the macroporous structure of the biocomposite scaffold and, upon further investigation it was recorded that the human osteoblast cells adhered to both the collagen and hydroxyapatite particle [Furthermore, bovine origin powdered and sintered hydroxyapatite was mixed with type I collagen to develop osteoinductive and osteoconductive scaffolds. The particle size in the scaffold varied between 200 and 400\u00a0nm. The surface-to-volume density and the volume density of the composite material varied from 5.090 to 6.366\u00a0\u00b5mparticle . Meat inparticle . Anotherparticle . Cow dunparticle .4.in vitro when turned into injectable forms [Pig waste based biomaterials have also found several applications in the biomedical industry. There are three major forms of prosthetic mesh. Synthetic meshes with high tensile strength, such as polypropylene (PP) or polyester, are expected to induce bowel adhesions, rendering them inappropriate for intra\u2212abdominal use. Composite meshes, also known as barrier meshes, comprise dual-sided prostheses with a synthetic parietal side facilitating robust healing and a visceral side that resists tissue ingrowth and reduces adhesion development. Biological meshes comprise of collagen-based scaffolds that could be implanted extra\u2212 or intra\u2212peritoneally . Biobranle forms .5.Hydroxyapatites (HAs) have good biocompatibity and are mostly utilized in bone replacement and tissue engineering . Hydroxy6.\u22121) depicted highest cell (fibroblast cells) adhesion, free radical scavenging activity and MTT activity in comparison to the DG-SIS and other nano bio-composite scaffolds. Thus, DG-SIS/NC3 scaffolds can be used as potential scaffold biomaterial for skin TE application. Collagens employed in tissue engineering nowadays are mostly from bovine or porcine sources. The potential of a spongiform encephalopathy epidemic has, however, limited the usage of collagen derived from these sources. Some scientists investigated the prospect of employing domestic goat accessible on the subcontinent as a viable supply of collagen for tissue-engineering applications, keeping the aforementioned perspective in mind. GTC (Goat tissue collagen) was discovered to be made up of type-I collagen. GTC improved cell adhesion, cell cycle progression, and proliferation, according to a biocompatibility study. Immunocytochemical study in combination with traction force microscopy demonstrated that the cell\u2013substrate connection in GTC is mediated by a superior focal adhesion complex. Finally, an in vivo investigation in mice indicated that GTC has low immunogenicity and dramatically improves the healing process. Calf skin collagen (CSC) was employed as a benchmark for comparison throughout the investigation. Therefore, GTC has the potential to be used as a biomaterial in skin tissue engineering [Among various animals used in manufacturing biomaterials, goat tissue is still underutilized in tissue engineering, although it is comparatively less susceptible to contamination or disease transmission than cadaveric porcine and bovine tissue. In a study, small intestinal submucosa (G-SIS) of goat was isolated out of goat\u2019s small intestine (G-SI), which is often a waste obtained from goat-slaughterhouse, and was decellularized in order to obtain decellularized G-SIS (DG-SIS) biomatrix in powder form, gel form and sponge form owing to its potential in healing different types of wounds. Nanoceria (NC) was induced in the DG-SIS in order to fabricate DG-SIS/NC nano bio-composite scaffold, which provides synergistic effects in rapid tissue regeneration. The scaffolds so obtained were hydrophilic, haemo-compatible, biodegradable, antibacterial, biocompatible and revealed free-radical scavenging capability. It was observed that the scaffolds containing higher NC concentration , 2,2-diphenyl-1-picrylhydrazyl (DPPH), peroxy and superoxide anion radicals at different protein hydrolyzate conditions [Apart from hoof-derived protein powders, protein hydrolyzates obtained from goat tannery fleshings have also been investigated for potential applications in antioxidant rich animal feed. These hydrolyzates after fermentation by  animals . Moreovenditions .7.Delftia acidovorans DSM39, but it cannot thrive on fatty substrates. A study was conducted to develop a D. acidovorans DSM39 recombinant strain capable of producing PHAs from waste fats such as lard, udder and tallow. Interestingly, the recombinant strain was able to obtain 43 and 39%, respectively, with almost 7% of 4HB from udder and fat, respectively. The findings of this study correspond to a one-step conversion of fatty residues into PHAs with beneficial properties that might be used in a wide range of industrial applications [ications . The useications . In otheications .Table 1.Dairy processing waste is another important resource that contains soluble organics, suspended solids, and trace organics , along with other essential components for microbial fermentation to produce biopolymers ,53. At pin vivo biodegradability [The blood from slaughterhouses has also been used to develop biocomposite scaffolds for muscle tissue. The generated scaffold exhibited elastic mechanical properties, a microporous structure, excellent cell proliferation and migration, and good dability .8.Animal processing and farm wastes contribute to biomaterial development and are generally derived from bones, hoofs, skin, and teeth. The production of biopolymers , composites, collagen, and other biomaterials from animal wastes, for biomedical and industrial applications was discussed in this review. Currently, biomaterials from animal waste are used for the fabrication of scaffolds, for promoting bone and tissue regeneration and for fibroblast cell growth. However, the method of biomaterial preparation lack standardized protocols in terms of pre-treatment, extraction, chemical modifications and purification that restrict its continuous commercial production. Process viability and optimization is required in this regard that should be achieved through pilot-scale investigations and not in a flask. Important parameters such as the crystallinity, hardness, porosity, surface area, swelling and elasticity, temperature and pH stability, cell viability, cytotoxicity/cytocompatibility, neurovascularization capacity, etc., should be investigated in-depth for the generated biomaterials. Moreover, the diversified production of biomaterials has not been fully explored utilizing animal waste. The comparative merit of biomaterial developed from animal waste should also be explored in contrast to biomaterials produced using other waste sources. Process economics and circular bioeconomy concepts also need to be developed for animal waste-derived biomaterials to make the industrial process of biomaterial fabrication more economical. Further, mere scientific interventions and industrial efforts may not be enough to successfully commercialize the production and distribution process of biomaterials generated from animal-derived waste. There should also be a continuous supply of resources to both research and industrial sectors to produce biomaterials more sustainably. For this, there is a need for government policy framework that would necessitate collecting animal-derived waste from the source and supplying it to the concerned industries and research organizations at subsidized charges. Such frameworks are the need of the hour that would promote both scientific advancement and national economy."}
+{"text": "Geothlypis trichas, Spotted Towhee Pipilo maculatus, Swainson's Thrush Catharus ustulatus, Song Sparrow Melospiza melodia, and Yellow Warbler Setophaga petechia). We used stable isotopes of \u03b413C and \u03b415N and Bayesian mixing models in MixSIAR to determine the reliance of riparian predators on aquatic prey sources. Using mixed effects models, we found that arachnid families varied in their reliance on aquatic prey sources. While songbird species varied in their reliance on aquatic prey sources, songbirds sampled earlier in the season consistently relied more on aquatic prey sources than those sampled later in the season. For both arachnids and songbirds, we found a positive correlation between the amount of the aquatic prey source in their diet and their Hg concentrations. While the seasonal pulse of aquatic prey to terrestrial ecosystems is an important source of nutrients to riparian species, our results show that aquatic prey sources are linked with higher Hg exposure. For songbirds, reliance on aquatic prey sources early in the breeding season (and subsequent higher Hg exposure) coincides with timing of egg laying and development, both of which may be impacted by Hg exposure.Cross\u2010ecosystem subsidies move substantial amounts of nutrients between ecosystems. Emergent aquatic insects are a particularly important prey source for riparian songbirds but may also move aquatic contaminants, such as mercury (Hg), to riparian food webs. While many studies focus on species that eat primarily emergent aquatic insects, we instead study riparian songbirds with flexible foraging strategies, exploiting both aquatic and terrestrial prey sources. The goal in this study is to trace reliance on aquatic prey sources and correlate it to Hg concentrations in common riparian arachnids  and songbirds . We determined that individual reliance on aquatic prey was positively correlated with mercury exposure, with songbirds relying more in aquatic prey early in the season. Our results show the seasonal pulse of aquatic prey to terrestrial ecosystems is likely an important source of nutrients to riparian species but may also drive season\u2010dependent exposure to aquatic contaminants. When assimilated by aquatic invertebrates, these contaminants can accompany their movement into surrounding terrestrial food webs . We use \u03b413C and \u03b415N stable isotopes to differentiate between aquatic and terrestrial signatures, which has been used in other studies with success  reliance on the aquatic prey source, 4) evaluate seasonal changes and species\u2010specific differences in the proportion of aquatic prey in the diet of terrestrial predators, and 5) correlate seasonal changes in aquatic prey reliance to their Hg exposure for both arachnids and songbirds.We hypothesize that some of the variation in Hg exposure among insectivorous predators (arachnids and songbirds) can be explained by species and individual\u2010level differences in prey selection, with individuals or species that rely more on aquatic prey having higher Hg exposure. Our goal in this study is to trace reliance on aquatic\u2010based prey and correlate it to Hg concentrations in common riparian arachnids  and songbirds , Spotted Towhee (Pipilo maculatus), Swainson's Thrush (Catharus ustulatus), Song Sparrow (Melospiza melodia), and Yellow Warbler (Setophaga petechia). All birds were banded with an aluminum USGS band, and any recaptures were excluded from the analyses to preserve independence of samples. Blood samples of each bird were taken from the brachial ulnar vein, using 27\u2010gauge needles  and heparinized microhematocrit capillary tubes . Samples were capped with Critocaps\u2122 (Leica Microsystems) and stored on ice in the field until they could be transferred to a freezer (within 6\u00a0hr of sampling). No more than 1% of bird's body weight of blood was collected from each individual, usually between 20\u00a0\u00b5l and 100\u00b5l.From 1 May 2013 to 23 July 2013, we sampled 7 main stem sites along the Willamette River Figure\u00a0. SingingWe recorded presence of brood patch  or cloacal protuberance  and limited samples to individuals in breeding condition to avoid late or early migrants. We also aged each bird Pyle,\u00a0 and checWe also collected aquatic and terrestrial invertebrates from sites that were collocated in space and time with the songbird sampling Table\u00a0. We samp2.2SD, minimum, and maximum number of individuals in the composite samples can be found in Table\u00a0Aquatic invertebrates were identified to family , ethylated with 1% sodium tetraethylborate, then analyzed via cold vapor atomic fluorescence spectrometry on a MERX\u2010M  automated methylmercury analyzer. Quality assurance measures included analysis of two independently derived liquid calibration standards, two certified reference materials . Percent recoveries averaged 99% (SD =8%) for 10pg MeHg standard, 96% (SD =13%) for 100pg MeHg standard, 99% (SD =21%) for IAEA\u2010452 averaged and 86% (SD =5%) for TORT\u20103.Methylmercury (MeHg) is the bioavailable form of Hg that biomagnifies through trophic levels; inorganic Hg does not biomagnify. Of the Hg in blood of most birds, 95%\u201399% is MeHg  for calibration verification, recovery of 98%  for certified reference material, and absolute percent difference of 2%  for duplicates.Songbird whole blood samples were not composited, but instead run on an individual basis as dry weight. Because 95%\u201399% of Hg in bird blood is MeHg  \u22121] *1,000 where R = the ratio of the heavy isotope to the light isotope. Nitrogen samples were standardized against N2 in air, and carbon isotopes were standardized against Vienna PeeDee Belemnite. Based on replicate analysis of standard reference materials, we calculated the instrument standard for invertebrates and bird blood separately. For invertebrates, reference materials included bovine liver , USGS\u201041 glutamic acid , nylon\u20105 . For bird blood, reference materials included bovine liver , USGS\u201041 glutamic acid , nylon\u20105 .Composited, dried, and homogenized invertebrates and dried whole blood from individual birds were packed in tin capsules for stable isotope analysis at the University of California, Davis Stable Isotope Facility. All samples were analyzed using a PDZ Europa ANCA\u2010GSL elemental analyzer interfaced to a PDX Europa 20\u201320 isotope ratio mass spectrometer . Stable isotope values are reported as delta (\u03b4) values using the equation \u2030 = [. We grouped invertebrates into broad categories based on diet and life history. Aquatic\u2010collected samples were grouped into aquatic , emergent , and emergent predators . Terrestrially collected samples were grouped into terrestrial , terrestrial\u2010emergent , terrestrial\u2010emergent predators , and terrestrial\u2010mixed . The terrestrial\u2010mixed category is necessary because we only identified terrestrial insects to order, and some of the orders have mixed life history strategies  followed by Tukey's HSD test to quantify differences in both \u03b413C and \u03b415N among the invertebrate groups. We also compared MeHg concentrations among the invertebrate groups (one\u2010way ANOVA on log\u2010transformed MeHg concentrations).To determine suitable stable isotope endmembers for our analysis, we first explored differences in \u03b4Based on these analyses, we designated the \u201cemergent\u201d insect group as being representative of the aquatic prey source and the \u201cterrestrial\u201d insect group as representative of the terrestrial prey source (groups explained above and in Table\u00a02.5.2N\u00a0=\u00a036) had mean \u03b413C\u00a0=\u00a0\u221221.937 (SD =2.707) and mean \u03b415N\u00a0=\u00a010.519 (SD =2.740). Terrestrial prey (N\u00a0=\u00a045) had mean \u03b413C\u00a0=\u00a0\u221228.019 (SD =1.553) and mean \u03b415N\u00a0=\u00a03.159 (SD =2.124). We chose trophic discrimination factors (TDF) based on available data for species with similar feeding habits and prey sources. We used different TDF for arachnids  and terrestrial sources  and songbirds . No TDF exists for the species sampled in this study, but we also ran our MixSIAR results using TDF for songbirds used in other studies  to estimate the proportion of the aquatic prey source in the diet of both riparian arachnids and songbirds. Prey sources supplied to MixSIAR were the same for both arachnid and songbird models. Aquatic prey . The songbird model included individual bird as a random effect, uninformative priors, a residual*process error structure and used the \u201cvery long\u201d run length to achieve model convergence . We assessed model convergence using Gelman\u2010Rubin and Geweke diagnostics.2.5.3We used the mixSIAR\u2010calculated proportion aquatic prey source for both arachnids and songbirds in all subsequent analysis. We developed mixed effects models using packages lme4 , or an interaction of date by group influenced proportion of aquatic prey of these arachnids. The interaction was not significant so we report the reduced model with only main effects. We then ran models to determine if proportion of aquatic prey and arachnid group influenced MeHg exposure, including site as a random effect.Second, for riparian songbirds, we ran a mixed effects model that accounted for site as a random effect to determine if date, songbird species , or an interaction of date and species influenced proportion of aquatic prey in the blood of riparian songbirds. The interaction was not significant, and so we report the reduced model with only main effects. We then ran a mixed effects model to determine if proportion aquatic prey, species, or an interaction of proportion aquatic prey and species influenced blood THg concentration in songbirds.33.113C, \u03b415N, and MeHg and 137 songbird individual samples for \u03b413C, \u03b415N, and THg ; their isotope signatures were similar to the aquatic environment. Overall and taxa\u2010specific means and SD for \u03b413C, \u03b415N, and geometric means and back\u2010transformed SE for MeHg can be found in Table\u00a013C  and \u03b415N  between invertebrate groups. Tukey pairwise analysis revealed consistent differences (p\u00a0<.01) between terrestrial insects and aquatic, emergent aquatic, and emergent aquatic predators in both \u03b413C and \u03b415N  indicated that terrestrial insects were significantly lower in MeHg than all other groups . Although there was no difference in \u03b413C or \u03b415N between aquatic invertebrates and emergent aquatic invertebrates, there was a significant difference in MeHg between these groups.Invertebrate groups also differed in MeHg concentrations  and terrestrial insects as our terrestrial prey source  for MixSIAR analysis for riparian arachnids and birds . We next used MeHg concentration as the dependent variable and found that MeHg concentrations also differed among arachnid groups  and was positively related to proportion aquatic prey . However, the interaction between proportion of aquatic prey and arachnid family  indicated that the slopes for the relationship between proportion of aquatic prey and MeHg concentrations varied among taxa  and was different among species . Throughout the season, proportion of aquatic prey in the songbird diet declined, from 31.5% (SE\u00a0=\u00a04.2%) at the early sampling period (Julian day 120 = Apr 30) to 11.3% (SE\u00a0=\u00a03.8%) at the late sampling point (Julian day 200 = July 19) . Proportion aquatic prey in bird diets was influenced by date  Figure\u00a0. Song Sp) Figure\u00a0.F\u00a0=\u00a027.9, p\u00a0<.0001) and differed among species , varying 3\u2010fold between the species with the lowest and highest concentrations. Swainson's Thrush  had the lowest least squares mean THg concentration, followed by Spotted Towhee , Yellow Warbler , Song Sparrow , and Common Yellowthroat   Figure\u00a0.413C and \u03b415N isotopes can be used to trace the amount of aquatic prey in the diet of riparian predators. We found that aquatic energy subsidies comprised a large component of riparian predator diets, and species in riparian habitats foraging on aquatic\u2010derived food items were more likely to have higher Hg exposure than those foraging on prey derived from terrestrial habitats. Contaminants like Hg are generally higher in aquatic than terrestrial environments; thus, these subsidies may represent substantial vectors of aquatic contaminants into terrestrial communities  or closely related conspecifics are widespread throughout North American riparian areas and represent an understudied community in aquatic contaminant studies. The species we studied varied in their reliance on aquatic prey and subsequent Hg exposure at both a species and individual level. We were not only able to correlate Hg concentrations in songbirds to their reliance on aquatic\u2010based prey, but also showed that birds forage on more aquatic\u2010sourced prey early in the season than later. These findings suggest that pulsed emergence of aquatic invertebrates may be an important vector of Hg to avian insectivores. These findings are particularly relevant in the face of climate change, which can alter the timing and magnitude of emergent aquatic subsidies ; Formal analysis (lead); Funding acquisition (supporting); Investigation (lead); Methodology ; Visualization (lead); Writing\u2010original draft (lead); Writing\u2010review & editing . Collin A. Eagles\u2010Smith: Conceptualization (supporting); Formal analysis (supporting); Funding acquisition (lead); Methodology ; Resources (lead); Supervision ; Writing\u2010review & editing . W Douglas Robinson: Conceptualization (supporting); Methodology (supporting); Supervision ; Writing\u2010review & editing ."}
+{"text": "Colorectal cancer (CRC) is a malignant tumor and morbidity rates are among the highest in the world. The variation in CRC patients' prognosis prompts an urgent need for new molecular biomarkers to improve the accuracy for\u00a0predicting the CRC patients' prognosis\u00a0or as a complement to the traditional TNM staging for clinical practice. CRC patients'\u00a0gene expression data of HTSeq\u2010FPKM and matching clinical information were downloaded from The Cancer Genome Atlas (TCGA) datasets. Patients were randomly divided into a training dataset and a test dataset. By univariate and multivariate Cox regression survival analyses and Lasso regression analysis, a prediction model which divided each patient into high\u2010or low\u2010risk group was constructed. The differences in survival time between the two groups were compared by\u00a0the Kaplan\u2013Meier method and the log\u2010rank test. The weighted gene co\u2010expression network analysis (WGCNA) was used to explore the relationship between all the survival\u2010related genes. The survival outcomes of patients whose overall survival (OS) time were significantly lower in the high\u2010risk group than that in the low\u2010risk group\u00a0both in the training and test datasets. Areas under the ROC curves which termed AUC values of our 9\u2010gene signature achieved 0.823 in the training dataset and 0.806 in the test dataset. A nomogram was constructed for clinical practice when we combined the 9\u2010gene signature with TNM stage and age to evaluate the survival time of patients with CRC, and the C\u2010index increased from 0.739 to 0.794. In conclusion, we identified nine novel biomarkers that not only are independent prognostic indexes for CRC patients but also can serve as a good supplement to traditional clinicopathological factors to more accurately evaluate the survival of CRC patients. We identified a 9\u2010gene signature screened out based on batch survival analysis that can be used not only as an independent prognostic index for CRC patients but also as a favorable supplement to traditional clinicopathological factors for clinicians to more accurately and effectively evaluate the survival of CRC patients.  For example, Bingrong et al identified an effective 6\u2010lncRNA signature with good performance that divides CRC patients into high\u2010and low\u2010risk groups that have significantly different survival times.In the present study, by a large CRC cohort from The TCGA database, we developed a credible expression\u2010based 9\u2010gene signature that divides patients into high\u2010and low\u2010risk groups with significant differences in OS. In the validation process, the signature in our study also reflected high reliability in the test dataset. Combined with clinicopathological factors such as the TNM stage, which has been used in the clinic, the signature can provide a more accurate prediction for individual patients during clinical work.22.1https://www.cancer.gov). A total of 530 samples was obtained in the aggregate dataset, including 42 normal tissue samples and 488 CRC tissue samples. After the exclusion of patients with incomplete survival information, a total of 467 patients with complete follow\u2010up data was included. A GEO dataset GSE103479 downloaded from the official website (https://www.ncbi.nlm.nih.gov/geo/) including 156 postoperative drug\u2010resistant stage II and stage III CRC samples was used during the further validating process  was conducted to screen the genes associated with patients\u2019 OS using the \u201csurvival\u201d package in R (version 3.6.0). Univariate Cox regression analysis was used to further identify genes that were screened by previous batch survival analysis associated with patients' OS and we set the filter criteria as HR>1.5 or HR<0.8 and p value\u00a0<\u00a00.01. Finally, 91 eligible genes were obtained. Then, we used the \u201ccaret\u201d package in R to randomly divide 467 samples into two groups, in which 235 samples were allocated to the training group and 232 samples were allocated to the test group.We downloaded the HTSeq\u2010FPKM gene expression data and corresponding clinical information of all postoperative CRC patients prior to October 2019 from the official website of the TCGA database  analysis based on the \u201cSurvivalROC\u201d R package\u00a0was used to assess the predictive stability and effectiveness of the 9\u2010gene signature in each group. To verify whether this 9\u2010gene signature can serve as an independent prognostic index compared with the existing traditional clinicopathological indicators for evaluating prognosis, we performed a multivariate regression analysis that takes into account our 9\u2010gene signature, TNM stage, age, sex, and so on. Finally, a nomogram and related calibration curves were established based on CRC patients for further clinical application.A multivariate Cox proportional hazards regression model was established and used to evaluate the pertinence between the expression value of the 9\u2010gene signature and CRC patients' OS. The risk score of each patient was constructed by weighting the expression value of prognostic genes according to their multivariate Cox regression coefficients. The formula is shown below:2.3To explore the relationship between the nine genes in our signature and all the survival\u2010related genes mentioned above, the weighted gene co\u2010expression network analysis (WGCNA) without clinical traits was analyzed. By selecting 5 as the soft threshold, a weighted gene co\u2010expression network was constructed by the R \u201cWGCNA\u201d package with approximate scale\u2010free properties. A highly synergistic matrix of genes was determined by the relevance among the expression values of all those genes. The network module was produced by the topological overlap measurement (TOM)2.4p\u00a0<\u00a00.05 was regarded as statistically significant. The statistical analysis was performed in R software (version 3.6.0).The survival\u2010related genes and survival differences between the high\u2010and low\u2010risk groups in the training dataset and test dataset were obtained by the \u201csurvival\u201d R package through the Kaplan\u2013Meier method and compared by the log\u2010rank test. Lasso regression analysis was performed by the \u201cglmnet\u201d R package to eliminate synergistic genes. The random grouping was completed by the \"caret\" R package. Time\u2010dependent ROC curve was used to describe the sensitivity and specificity of survival prediction based on the risk score generated by the \u201csurvivalROC\u201d R package. Multivariate Cox regression analysis was used to show the independent predictability of the signature and generate a nomogram. The calibration curve was used to evaluate whether patients' actual survival was consistent with\u00a0that predicted by the nomogram. The nomogram and calibration curve were generated by the \u201crms\u201d R package. The \u201cpec\u201d R package was used to compare the concordance index between our nomogram and nowadays TNM stage added with age through C\u2010index function. All statistical tests were two\u2010sided and 33.1p value lower than 0.05. Setting the filtered standard as HR>1.5 or HR<0.8 and p value <\u00a00.01, 91 genes were selected for further research. A total of 467 CRC patients was divided randomly into two groups: a training group (n\u00a0=\u00a0235) and a test group (n\u00a0=\u00a0232). Univariate Cox regression analysis was used again to verify the effect of the 91 genes on the OS of CRC patients in the training group and 62 genes achieved a statistical significance. To improve the interpretability and prediction accuracy of the regression model and to solve the problem of collinearity of variables, we performed Lasso regression analysis of the 62 genes in the training dataset and the results showed that 17 genes were screened out as the basis for our further construction of a model to predict the OS of CRC patients  and risk score (high vs. low)  could be used as independent prognostic factors  and clinicopathological factors such as age, sex, and TNM stage. The results showed that stage  to 0.794 (95% CI\u00a0=\u00a00.737\u20100.851), indicating an obvious increase in C\u2010index after adding our signature Figure\u00a0. Calibra3.5To explore the possible co\u2010expression relationship between the genes in our signature and other survival\u2010related genes, we performed WGCNA without clinical traits. With a scale\u2010free network and topological overlaps, by selecting 5 as the soft threshold and merging similar modules, we generated a hierarchical clustering tree, and 19 corresponding gene modules were identified Figure\u00a0, in whic4CRC is a disease with high intercellular heterogeneity,Of the nine genes  in our signature, researchers have proven that SALL1 can act as a tumor suppressor by recruiting NuRD\u00a0to induce tumor cell senescence in breast cancer during which it is controlled by\u00a0the MAPK and mTOR signaling pathways.The 9\u2010gene signature can classify CRC patients into high\u2010and low\u2010risk groups with a significant difference in survival time. It is worth noting that when we tried to prove the reliability of this 9\u2010gene signature, the results of ROC analysis proved that the AUC values in both the training dataset and test dataset were greater than 0.80 and in the validation dataset it was 0.651, indicating that the nine\u2010gene combination can be regarded as a reliable and efficient prognostic indicator for CRC patients. Further comparison in the validation dataset showed that the 9\u2010gene signature was better than Zhou's 5\u2010gene signature and had an equivalent efficiency as the TNM stage in predicting CRC patients\u2019 5\u00a0years\u2019 OS time verified by AUC values. It should be mentioned that the validation dataset is a collection of drug\u2010resistant stage II and stage III CRC patients, which suggest that our signature may also benefits drug\u2010resistant CRC patients with a specific treatment based on the risk score calculated on those nine genes. What is more, the multivariate Cox regression analysis showed that it was independent of other clinical factors, such as age, sex, and stage. TNM stage is the current gold standard for evaluating the prognosis of CRC patients.In conclusion, the 9\u2010gene signature screened out based on batch survival analysis can be used not only as an independent prognostic index for CRC patients but also as a favorable supplement to traditional clinicopathological factors for clinicians to more accurately and effectively evaluate the survival of CRC patients.The authors declare that they have no competing interests.Figure S1Click here for additional data file.Figure S2Click here for additional data file.Figure S3Click here for additional data file.Figure S4Click here for additional data file.Figure S5Click here for additional data file.Figure S6Click here for additional data file.Supplementary MaterialClick here for additional data file."}
+{"text": "Glioblastoma multiforme (GBM) is a highly, malignant tumor of the primary central nervous system. Patients diagnosed with this type of tumor have a poor prognosis. Lymphocyte activation plays important roles in the development of cancers and its therapeutic treatments.We sought to identify an efficient lymphocyte activation-associated gene signature that could predict the progression and prognosis of GBM.n\u00a0=\u00a0525). Then, the signature was validated in two datasets, including GSE16011 (n\u00a0=\u00a0150) and GSE13041 (n\u00a0=\u00a0191) using the Kaplan Meier method. Univariate and multivariate Cox proportional hazards regression models were used to adjust for clinicopathological factors.We used univariate Cox proportional hazards regression and stepwise regression algorithm to develop a lymphocyte activation-associated gene signature in the training dataset  in the training dataset and classified the patients into high-risk and low-risk groups with significant differences in overall survival . This signature showed similar prognostic values in the other two datasets. Further, univariate and multivariate Cox proportional hazards regression models analysis indicated that the signature was an independent prognostic factor for GBM patients. Moreover, we determined that there were differences in lymphocyte activity between the high- and low-risk groups of GBM patients among all datasets. Furthermore, the lymphocyte activation-associated gene signature could significantly predict the survival of patients with certain features, including IDH-wildtype patients and patients undergoing radiotherapy. In addition, the signature may also improve the prognostic power of age.We identified a lymphocyte activation-associated gene signature (In summary, our results suggested that the lymphocyte activation-associated gene signature is a promising factor for the survival of patients, which is helpful for the prognosis of GBM patients. Glioblastoma multiforme (GBM) is classified as a grade IV diffuse glioma and is one of the most aggressive and lethal brain cancers. GBM has a high recurrence rate, typically originates in the cerebral hemispheres, and can quickly spread to the other parts of the brain\u00a0. IDH staThe lymphocyte activation is a set of processes where the lymphocytes are stimulated by specific antigens or nonspecific mitogens, which results in the synthesis of protein and production of lymphokines\u00a0. These pIn our study, a lymphocyte activation-associated gene signature was developed, which predicted the overall survival of GBM patients in training dataset. And we validated its prognostic power in another two datasets. The signature was found to be an independent prognostic factor after adjusting for other clinicopathologic factors. Moreover, this signature not only predicted the survival of patients with IDH wild-type glioblastoma, but also patients after radiotherapy. These findings indicated the signature may serve as an effective prognostic biomarker for patients with GBM.GSE16011\u00a0 and Gene Expression Omnibus (GEO)\u00a0(GSE13041 and 150 GBM patients from GSE16011) were included in this study. The TCGA dataset was treated as a training dataset, and another two datasets (GSE16011 and GSE13041) were used as independent validation cohorts. In the training dataset, the median survival time of GBM patients was 12.4 months (ranging from 0.1 to 127.5 months). And there were 144 classical, 155 mesenchymal, 83 neural, and 99 proneural GBM samples. In We downloaded three gene expression datasets  from Thus (GEO)\u00a0, which cP < 0.05). We focused on the prognostic genes that participated in the lymphocyte activation process. The lymphocyte activation-related genes were collected by GO terms in Gene Ontology\u00a0 that were identified in our study (TCF3) + (0.2992\u00a0\u00a0\u00d7 expression level of IGFBP2) + (0.2421068\u00a0\u00a0\u00d7 expression level of TYRO3) + (0.2155469\u00a0\u00a0\u00d7 expression level of NOD2). Based on this model, the patients were classified into the high- and low-risk groups by the median risk score in the training dataset.Differential expression analysis was performed based on transcriptome profile of GBM patients by the \u201climma\u201d R package in the training dataset. Genes with the cutoff criteria of \u2014log2-fold change\u2014\u22651 and FDR < 0.000001 between tumor and normal tissues were regarded as differentially expressed genes (DEGs). Using the univariate Cox proportional hazards regression algorithm, we retained the DEGs that were strongly associated with the survival of patients with GBM  of lymphocytes  from the CIBERSORT web portal\u00a0 was used to perform all of the statistical analyses, version 3.5.1 .In the survival analysis, we analyzed only overall survival as the end point. The survival differences between the two groups were visualized by Kaplan\u2013Meier analysis and were compared by the log-rank test. The prognostic value of the lymphocyte activation-associated gene signature was estimated by univariate and multivariate Cox proportional hazards regression models. The Cox proportional hazards regression model was used to calculate hazard ratio (HR) and 95% confidence intervals (CI). The b portal\u00a0 and MCP-b portal\u00a0. We combb portal\u00a0 based onTCF3, IGFBP2, TYRO3 and NOD2) that were significantly related to overall survival using a stepwise regression algorithm. We used the lymphocyte activation-associated gene signature to calculate each patient\u2019s risk score based the four genes\u2019 expression levels weighted by regression coefficients in the univariate Cox proportional hazards regression analysis  suggested that their expression was associated with poor prognosis in GBM patients, while the negative coefficient of TCF3 (\u22120.6246210) indicated it was associated with better survival.The gene expression profile and clinical information of GBM patients were obtained from TCGA, which were treated as the training dataset. There were 525 tumor samples and 10 normal samples. The tumor and normal samples came from different samples. We identified 2,364 differentially expressed genes between tumor and normal samples by using the limma package (FDR <\u00a00.000001 and \u2014log2-fold change\u2014 \u22651), including 933 up-regulated and 1,431 down-regulated genes . We obtaanalysis . Positivn\u00a0=\u00a0262) and low-risk (n\u00a0=\u00a0263) groups according to the median risk score (2.752) of the lymphocyte activation-associated gene signature. This score was used as the cutoff. Patients within high-risk group had significantly shorter overall survival time than those within low-risk group  and GSE16011 (n\u00a0=\u00a0150). Each patient\u2019s risk score was calculated with the same formula as that used in the training set. The patients were divided into the high- and low-risk groups according to the median cutoff determined for each dataset. In GSE16011, survival time of patients in the high-risk group were significantly shorter than those in the low-risk group , age , sex  and proneural types  independently predicted a worse OS for GBM patients in the training dataset . These lymphocytes were memory B cells, naive B cells, naive CD4 T cells, follicular helper T cells and regulatory T cells  but it did not predict the survival of IDH-mutation patients  among IDH-wildtype patients . In addition, the signature did not have the prognostic ability for patients with neural and classical subtypes  established by Verhaak et al. using Kaplan\u2013Meier survival analysis. The signature significantly predicted the overall survival of patients with the mesenchymal and proneural subtypes  and GSE13041 datasets  and between age and sex. The clinical factors for age in combination with the signature showed a higher C-index  than age or histological grade alone . These results suggested that this lymphocyte activation-associated gene signature could add complementary value to known clinical factors.To evaluate the prognostic performance of the prognostic signature, we performed C-index calculation of clinical factors (age and sex) and the signature in three datasets\u00a0. We consIn this study, we identified a lymphocyte activation-associated gene signature that predicted shorter overall survival of patients with GBM. The prognostic power of this signature was evaluated by univariate and multivariate Cox proportional hazards regression models analysis using three datasets (HR >\u00a01). Our findings indicated that the gene signature was an unfavorable factor in GBM. Moreover, the high-risk and low-risk patients, which were separated by the signature, showed significant differences in immune cell activity. This signature was found to be an independent prognostic factor after adjusting for certain clinicopathological factors. In addition, the lymphocyte activation-associated gene signature also predicted the survival of patients with an IDH wild-type variant and for patients after radiotherapy treatment.TCF3, IGFBP2, TYRO3 and NOD2). IGFBP2 was one of the insulin-like growth factor binding proteins (IGFBPs), which are proteins binding to Insulin-like growth factors. IGFBP2 expression increased in peripheral blood mononuclear cells and participated in lymphocyte proliferation\u00a0(IGFBP2 can be as potential biomarker and therapeutic target\u00a0(IGFBP2 expression predicted the survival in GBM patients\u00a0(NOD2 played important roles in the pathogenesis of some diseases, such as, oral lichen planus\u00a0(NOD2 expression is higher in activated/memory CD4+ T cells\u00a0(We developed a prognostic signature that included four genes (feration\u00a0. In varic target\u00a0. Moreovepatients\u00a0. NOD2 pln planus\u00a0, Crohn\u2019sn planus\u00a0, and infn planus\u00a0. NOD2 ex T cells\u00a0 and was  T cells\u00a0. A progn T cells\u00a0.TCF3 played a role in germinal center B - cell development and promoted cell growth, which contributed to proliferative phenotype in Burkitt lymphoma\u00a0(TCF3 can regulate B-cell-restricted genes through E-box motifs\u00a0(TCF3 promotes the survival in lymphoid cells\u00a0(TYRO3 is a protein-coding gene, which participates in ERK signaling pathway. The higher level of TYRO3 expression is associated with decreased overall survival in patients with colorectal, hepatocellular, and breast cancers\u00a0(Another marker, an important transcription factor lymphoma\u00a0. In addix motifs\u00a0. It can x motifs\u00a0. Also, Tid cells\u00a0. TYRO3 i cancers\u00a0. AlthougIn conclusion, we developed a lymphocyte activation-associated gene signature with prognostic power and offered new insights for the treatments of GBM. However, more data are needed to test the prognostic value of the signature before applying it to clinical management. A larger study is needed to confirm that the signature can accurately predict the prognostic benefits for GBM patients.10.7717/peerj.12070/supp-1Supplemental Information 1Click here for additional data file."}
+{"text": "Cancer is considered the second leading cause of death worldwide and in 2018 it was responsible for approximately 9.6 million deaths. Globally, about one in six deaths are caused by cancer. A strong correlation was found between diabetes mellitus and carcinogenesis with the most evident correlation was with type 2 diabetes mellitus (T2DM). Research has proven that elevated blood glucose levels take part in cell proliferation and cancer cell progression. However, limited studies were conducted to evaluate the efficiency of conventional therapies in diabetic cancer patients. In this review, the correlation between cancer and diabetes will be discussed and the mechanisms by which the two diseases interact with each other, as well as the therapeutics challenges in treating patients with diabetes and cancer with possible solutions to overcome these challenges. Natural products targeting both diseases were discussed with detailed mechanisms of action. This review will provide a solid base for researchers and physicians to test natural products as adjuvant alternative therapies to treat cancer in diabetic patients. Diabetes mellitus is a class of metabolic disorders characterized by prolonged periods of hyperglycemia. According to the World Health Organization (WHO), in the year 2014, 422 million people worldwide suffered from diabetes, and 1.6 million deaths were directly related to diabetes in the year 2016. Moreover, these statistics have been rising slowly over the last few decades . Studies9 cells  if suitable conditions are available  is an isoflavonoid derivative and it belongs to phytoestrogen classes. It exists in glycosylated or free forms in food (mainly legumes). It was firstly isolated from activity ,218. Theactivity ,220. Genactivity . Genisteactivity .A study stated that genistein has anti-inflammatory and antidiabetic functions, especially direct effects on the proliferation of \u03b2-cells and the secretion of insulin. Genistein improved glucose homeostasis . In \u03b2-ceTanacetum parthenium (feverfew) [Tanacetum vulgare (tansy) and Tanacetum larvatum [In the Asterceae family of medicinal plants, especially in everfew) , partheneverfew) . It can larvatum . Parthenlarvatum . Parthenlarvatum , STAT3 ilarvatum , p.53 aclarvatum . A studylarvatum . Parthenlarvatum , becauselarvatum . Furtherlarvatum . In panclarvatum .Parthenolide can reduce inflammation and remodel the impaired insulin signaling pathway, allowing cubilin and albumin uptake to be expressed . During Platycodon grandiflorum, Perilla frutescens, Apium graveolens, Cajanus cajan, Apium graveolens, Mentha spicata, and Sesbania grandifolra [Luteolin is a natural flavonoid abundantly present in many plant species. It is mainly found in vegetables and fruits, such as sweet bell peppers, carrots, onion leaves, broccoli, parsley, chrysanthemum flowers, and celery ,237. It ndifolra . Luteolindifolra . It has ndifolra . It obstndifolra ,241,242.ndifolra . The extndifolra . Stimulandifolra . Furtherndifolra .Colon cancer and diabetes are the primary cause of death worldwide . VariousAesculus hippocastanum, Ginkgo biloba, and Hypericum perforatum [Quercetin is a polyphenolic flavonoid found in apples, berries, broccoli, onions, broccoli, green tea, red tea , Aesculurforatum . The biorforatum . Quercetrforatum . It indurforatum . Moreoverforatum ,254. Querforatum . It alsorforatum . A studyrforatum .Quercetin can decrease serum glucose level through several mechanisms, including antioxidant action, modulating hepatic gene expressions, obstructing \u03b1-glucosidase activity in vitro, and improving insulin action along with skeletal muscle mitochondrial biogenesis enhancement . It was Coptis japonica Makino, Coptis chinensis Franch, and Berberis aristata Sims is an essential isoquinoline alkaloid. It is found in barberry, goldenseal, tree turmeric, and oregon grape [Berberine isolated from on grape . It is aon grape .Colorectal, lung, liver, ovarian, prostate and cervical cancers are all inhibited by berberine . Under iBerberine decreased blood glucose and the risk of metabolic syndrome, improved insulin sensitivity, stimulated weight loss, improved lipid metabolism, reduced the levels of hemoglobin A1C and triglyceride, increased the mRNA expression of adiponectin, and reduced leptin and resistin secretion . ReductiPhytosterols are plant-derived lipid compounds that are similar to cholesterol but differ in their carbon side chains and the presence or absence of a double bond. They are divided into two categories which are sterols (unsaturated compounds) and stanols (saturated molecules) . PhytostPhytosterols can be utilized to improve breast, ovary, liver, lung, prostate and stomach cancers. They have several suggested mechanisms, including inhibition of carcinogen production, angiogenesis, cancer cell growth, multiplication, invasion, and metastasis and induction of apoptosis and cell cycle arrest . Female Phytosterols have been shown to substantially lower total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) in the blood, and rise the HDL-C/LDL-C and HDL-C/TC indexes . ErgosteResearch and studies about the role of reactive oxygen species (ROS) and the effect of exogenous administration of antioxidants\u2019 supplements in cancer are opposed with controversy and conflicting outcomes. On one hand, antioxidants are commonly used by cancer patients and healthy individuals as a cancer-fighting strategy ,270,271.N-acetylcysteine (NAC) and vitamin E noticeably enhances human lung cancer cells proliferation and tumor progression in mice with B-RAF- and K-RAS-induced lung cancer via decreasing ROS, DNA damage, and p53 levels. Additionally, Volkan S. et al. showed that the antioxidants supplementation may accelerate early tumors or precancerous lesions growth in elevated-risk populations (like smokers and chronic obstructive pulmonary disease patients who receive NAC to reduce mucus production) because the somatic p53 mutations occur late in the tumor progression process [\u2212/\u2212Il2rg (NSG) mice) when supplemented regularly with subcutaneous injection of NAC [Klein A et al. conducted a study between 2004 and 2011 and found that prostate cancer risk for healthy men was markedly increased upon supplementation with 400 IU/day of vitamin E supplement , whereas process . Another process . Moreove process . Indicat process . Similarn of NAC . ClinicaIn conclusion, the supplementation of exogenous antioxidants may aid cancer cells to navigate the metastatic cascade and adopt adaptive steps to counteract the beneficial effect of antioxidant supplements. Nevertheless; additional clinical studies are required to confirm the generalizability of these remarks and to expand them to practical application especially with existing cancer patients.Cancer and diabetes represent global health challenges and the number of patients is contentiously increasing for both diseases. Hyperglycemia, hyperinsulinemia, and imbalanced fat metabolism in diabetic patients increase the altered metabolism in cancer cells. This metabolic link between cancer and diabetes complicates therapeutic protocols and reduces survival rates. Natural products with their diverse mechanisms of action can be used as adjuvant therapy to enhance conventional treatments. These natural products act by targeting interconnected mechanisms between cancer and diabetes. Further research is needed to explore the potential use of natural products in combination therapies to treat cancer in diabetic patients."}
+{"text": "Bone disease is among the defining characteristics of symptomatic Multiple Myeloma (MM). Imaging techniques such as fluorodeoxyglucose positron emission tomography\u2013computed tomography (FDG PET/CT) and magnetic resonance imaging (MRI) can identify plasma cell proliferation and quantify disease activity. This function renders these imaging tools as suitable not only for diagnosis, but also for the assessment of bone disease after treatment of MM patients. The aim of this article is to review FDG PET/CT and MRI and their applications, with a focus on their role in treatment response evaluation. MRI emerges as the technique with the highest sensitivity in lesions\u2019 detection and PET/CT as the technique with a major impact on prognosis. Their comparison yields different results concerning the best tool to evaluate treatment response. The inhomogeneity of the data suggests the need to address limitations related to these tools with the employment of new techniques and the potential for a complementary use of both PET/CT and MRI to refine the sensitivity and achieve the standards for minimal residual disease (MRD) evaluation. Multiple Myeloma (MM) is a plasma-cell malignancy in which a single clone of plasma cells proliferates and produces a monoclonal protein (MC). Myeloma clonal plasma cells accumulate in the bone marrow (BM), resulting in diffuse skeletal involvement, hypercalcemia, anemia and extramedullary localizations ,2,3,4. I18F-Fluorodeoxyglucose (FDG)-PET/CT is considered the best functional imaging technique in defining the metabolic activity of bone lesions, plasmacytomas and extra-medullary disease (EMD) caused by MM. 18F-FDG is currently the most used tracer for MM and it is a radiolabeled glucose analogue in which the C-2 hydroxyl group is replaced by the positron-emitting fluorine-18 atom (18F) [18F-FDG-6P, that is stored in cells as it cannot be further metabolized. After intravenous administration, 18F-FDG reaches an equilibrium state in 60 min and it starts to decay in 80\u201390 min, allowing an accurate assessment of glucose metabolic activity. MM plasma cells usually overexpress hexokinase-2, displaying a higher glycolytic activity than surrounding cells [18F-FDG uptake higher than the liver uptake, that is taken as a background with a Deauville score of 4. Consequently, PET positive lesions have a Deauville Score of 4 or 5 [18F-FDG-PET/CT is used to clarify a dubious diagnosis between MM and smoldering multiple myeloma (SMM), when low-dose WB-CT and WB-MRI are inconclusive but there is a strong suspicion of active MM, because of the high cost of this imaging technique. 18F-FDG-PET/CT is also used for the diagnosis of suspected extramedullary solitary plasmacytoma [18F-FDG-PET/CT is in the assessment of treatment response. After the first line therapy, patients undergo a repeat bone marrow (BM) aspiration, for the evaluation of MRD, and 18F-FDG-PET/CT is the only imaging technique able to distinguish between active and inactive FLs, with high prognostic value: 18F-FDG uptake in bone lesions has higher sensitivity in detecting tumor residual disease than immunofixation electrophoresis [18F-FDG-PET/CT Deauville criteria for MM, Zamagni et al. defined the complete metabolic response as the 18F-FDG uptake minus the liver activity in BM sites and FLs previously involved (including extramedullary and para-medullary disease (DS score 1\u20133)) and partial metabolic response as the decrease in number and/or activity of BM/FLs present at baseline, but persistence of lesion(s) with an uptake greater than the liver activity (DS score 4 or 5). The metabolic disease is defined as stable if there are no significant changes in BM/FLs and progressive when new FLs appear compared with baseline [om (18F) . FDG is ng cells . PET posf 4 or 5 . For a bf 4 or 5 . 18F-FDGmacytoma , EMD andmacytoma . These tmacytoma . The maiphoresis  and appebaseline .18F-FDG-PET/CT in bone lesions\u2019 detection at diagnosis ranges from 59% to 100%, whereas the specificity ranges from 75% to 82% [11C and 18F, becoming more sensitive than 18F-FDG [18F-FDG [11C-methionine is more sensitive than 18F-FDG and 11C-choline [18F-fluciclovine, a leucine analogous, is also a promising tracer for MM, reaching a similar uptake pattern and similar sensitivity, with a half-life of 110 min [18F-fluoro-ethyl-tyrosine could be used as a tracer, but it is less sensitive than 11C-metionine and 18F-FDG, having minimal transportation into plasma cells [11C-thiothymidine sensitivity is similar to the 11C-methionine one and it is better than 18F-FDG sensitivity, especially during the early stage of the disease [18F-fluorothymidine was investigated as a new tracer, but it is not considered a useful biomarker for MM [18F-fluoride reflects the early phase of bone calcification, but results concerning the diagnostic assessment in MM are divergent [89Zr-bevacizumab, a radiolabeled antibody directed to the VEGF receptor, is uniformly expressed on plasma cells and it could be useful for the detection of MM [18F-FDG is not clear, but a positive 68Ga-pentixafor has a negative prognostic significance, with a poorer OS [The sensitivity of % to 82% . False p% to 82% . False n% to 82%  and in c 18F-FDG ,27. Inde[18F-FDG ,29. Amin-choline , even if-choline . Stokke  110 min . 18F-fluma cells . Nucleos disease . 18F-flur for MM . 18F-fluivergent . 89Zr-beon of MM . In the oorer OS . CD38 isDue to the patchy dissemination of the disease, MRD evaluation in the BM could sometimes lead to false negative results in the presence of minimal disease after therapy . In the 18F-FDG-PET/CT leads to a more accurate assessment of response after a treatment line with the highest prognostic value, well related to PFS and OS. In fact, although MRD-negativity is associated with improved outcomes, in MRD-negative patients relapse still occurs, potentially due to the presence of focal bone disease that could be detected with 18F-FDG-PET/CT. The IFM 2009 trial showed patients who were double negative  achieving better PFS than patients who were not double negative [18F-FDG-PET/CT evaluation at diagnosis and one month after the end of the treatment. It was observed that patients MRD-/PET- had the best 4-years OS (94.2%) and PFS (92 months), patients MRD+/PET\u2212 had a not significant difference in 4-years OS (100%) but a shorter PFS (45 months) and PET+ patients had the worst 4-years OS (73.8%) and PFS (28 months) [18F-FDG-PET/CT positivity after first line treatment is the most affecting factor on PFS and OS.MRD negative is a deeper level of response than a complete response, that requires the absence of phenotypically abnormal clonal plasma cells from BM aspirates, detected by next-generation flow cytometry (NGF) or next-generation sequencing (NGS). However, MM is a heterogenous disease with patchy infiltration and EMD is not uncommon. The association between BM analysis and negative . Alonso  months) . This st18F-FDG-PET/CT in the therapy response assessment, even if they are not exactly concordant at the time of evaluation and in the interpretation of results. The main disadvantages of the 18F-FDG-PET/CT, beyond the high cost and the limited availability, are the lack of standardization criteria and of interobserver reproducibility in the interpretation of results. More than 30 clinical studies on the prognostic role of PET/CT in the evaluation of treatment response were found. In Numerous studies evaluated the accuracy of After therapy, the sensitivity of BM samples is reduced because marrow involvement in MM could be patchy, while MRI shows a better assessment for detecting diffuse marrow involvement, correlating with response to therapy . WB-MRI WB-MRI can be divided into anatomical MRI sequences, fat and water fraction sequences, diffusion-weighted imaging (DWI) and dynamic contrast-enhanced (DCE) MRI. A typical MRI pattern in MM is characterized by hypo-intensity and high contrast-enhancement in T1-weighted, hyper-intensity in T2-weighted and signal loss in the opposite phases . MRI canWB-MRI typical protocols are various. Anatomic sequences such as T1- and T2-weighted imaging could be used for identification and measurement of focal marrow replacing lesions . The DixIn conclusion, functional MRI sequences help to better characterize FLs and they have a prognostic value in MM; in particular, DWI is a promising tool for response assessment after treatment in this setting.NGF with functional imaging has been used to define responses in MM , though Although IMWG guidelines indicate the use of DWI-MRI to evaluate plasma cell disorders, the use of MRI to assess response to treatment is a matter of debate and it is investigated in several clinical studies, in small series of patients. In recent years, some studies have researched the role of DWI in treatment response assessment in patients with MM . They haDespite these current data, a better definition of the DWI-WB-MRI role in treatment assessment is necessary. Nowadays some prospective studies are ongoing to determine the sensitivity, specificity and the accuracy of WB-MRI. Additionally, some investigators are studying in MM patients the Dual-Echo T2 weighted acquisition for Enhanced Conspicuity of Tumors, an alternative WB-MRI technique that improves tumor visualization overcoming the compromised image quality of DWI-MRI (NCT04493411).Evidence shows that both MRI and PET/CT have significant potential in evaluating treatment response as separate tools. Specifically, data from PET/CT studies point out its prognostic and predictive value  whereas Several studies have also investigated which modality performs better in the setting of treatment-response evaluation. A recent comparative meta-analysis of Yokoyama et al. has shown the major impact of PET/CT rather than MRI in response to therapy  . The proIndeed, the studies reviewed in these articles display controversial data for techniques comparison underlining the need to understand and address MRI and PET/CT limitations .Concerning comparison of PET/CT and functional MRI for MRD assessment, clinical studies are few. According to IMWG, PET/CT represents the best tool for MRD evaluation as it has been shown to predict survival after therapy. However, most clinical trials using PET/CT as MRD tool are still ongoing and preliminary data do not show significant concordance between imaging and BM assessment . A recenEven though recommendations for the use of different techniques may vary depending on available local resources, we personally use PET/CT at diagnosis and at the end of a fixed treatment therapy (3 months after ASCT) and then every 6 months for 2 years. We integrate radiological study with MRI when PET is negative for disease and when local vertebroplasty or radiotherapy is needed. 18F-FDG-PET/CT yields false negative results losing sensitivity [In conclusion, in agreement with current data coming from comparative studies, PET/CT is probably the best choice for the evaluation of treatment response, bringing also prognostic value. A condition where DWI-WB-MRI sensitivity overcomes FDG-PET is disease with low expression of hexokinase-2, involved in the glycolytic pathway, as sitivity . In addisitivity ,89. To gsitivity . Furthersitivity . Combina"}
+{"text": "Recurrent multifocal adult rhabdomyoma in an elderly woman diagnosed with Birt-Hogg-Dub\u00e9 syndrome: A case report By Ulrik A, Marie R and Preben H. (2022) Front. Surg. 9: 1017725. doi: 10.3389/fsurg.2022.1017725A corrigendum on Error in Figure/TableIn the published article, there was an error in The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.Reminder: Figures, tables, and images will be published under a Creative Commons CC-BY licence and permission must be obtained for use of copyrighted material from other sources . It is the responsibility of the authors to acquire the licenses, to follow any citation instructions requested by third-party rights holders, and cover any supplementary charges."}
+{"text": "Azospirillum brasilense, Pseudomonas geniculata and their co-inoculation on growth and metabolic aspects of flax (Linum usitatissimum) plants that already grown in salinized soil.Salinized soils negatively affect plant growth, so it has become necessary to use safe and eco-friendly methods to mitigate this stress. In a completely randomized design, a pot experiment was carried out to estimate the influence of the inoculation with endophytic bacterial isolates A. brasilense and P. geniculata  increases almost growth characteristics . Moreover, contents of chlorophylls and carotenoids pigments, soluble sugars, proteins, free proline, total phenols, ascorbic acid, and potassium (K+) in flax plants grown in salinized soil were augmented because of the inoculation with A. brasilense and P. geniculata. Oppositely, there are significant decreases in free proline, malondialdehyde (MDA), hydrogen peroxide (H2O2), and sodium (Na+) contents. Regarding antioxidant enzymes, including superoxide dismutase (SOD), peroxidase (POD), and ascorbate peroxidase (APX), the inoculation with the tested endophytes led to significant enhancements in the activities of antioxidant enzymes in stressed flax plants.The results observed that inoculation of salinity-stressed flax plants with the endophytes Azospirillum brasilense, Pseudomonas geniculata  could be regarded as an uncommon new model to alleviate salinity stress, especially in salinized soils.The results of this work showed that the use of the endophytic bacterial isolates  The life of plants is constantly exposed to inappropriate conditions and various environmental stresses such as salinity, drought, chilling, heat, heavy metals, gaseous pollutants, pathogens, and other conditions that cause deleterious effects on the growth, metabolism, and productivity of various crops  is a dicotyledonous plant that belongs to the family Linaceae. It is an economically valuable fiber crop and oilseed (linseed) plant grown all over the world, being it an important source of natural fibers and industrial oil, and can meet the needs of edible oil and protein deficiency . From this point, our study was conducted to evaluate the possible role of these endophytes in salinity tolerance enhancements of flax plants by measuring some parameters such as growth aspects, photosynthetic pigments, soluble sugars, soluble proteins, free proline, total phenols, ascorbic acid, minerals (Na+ and K+), the activity of antioxidant enzymes , lipid peroxidation, hydrogen peroxide contents.Flax . About 1\u00a0ml from each dilution of intercellular fluid of its tissue was plated in triplicate and kept in an incubator at 28\u00a0\u00b0C for 48\u00a0h. One of them was Azospirillum brasilense, which was isolated from the stem of a cultivated-barley plant in Ras-Sudr, South of Sinai, Egypt. While the other isolate was Pseudomonas geniculata strain, which was isolated from the stem of a cultivated-wheat plant in El-Maghra region\u2014North of Sinai, Egypt. Depending on the molecular level, both strains were identified using a partial 16S rRNA gene sequence technique according to Berg et al.   in the rhizosphere soil were determined according to Parmer and Schmidt  seeds were obtained from Agricultural Research Center (ARC), Giza, Egypt. Microbial inoculants of A. brasilense, P. geniculata, and co-inoculation of both strains were used for treating flax plants. The present experiment was performed in a completely randomized design (CRD) with four replicates; the treatments and its replicates were randomly distributed within the study area. Ten seeds of flax crops were planted in pots (30\u00a0cm in diameter) containing 10\u00a0kg of salinized clay soil obtained from Sahl El-Tina, Sinai, Egypt. The characteristic features of the collected soil were as follows: soil depth 0\u201330\u00a0cm\u22121, texture clay, E.C. 11.3 dS/m, pH 7.4, HCO3\u2212 17.93\u00a0mg\u00a0g\u22121, Cl\u2212 51.03\u00a0mg\u00a0g\u22121, \u22121, Ca2+ 20.53\u00a0mg\u00a0g\u22121, Mg2+ 14.53\u00a0mg\u00a0g\u22121, Na+ 55.63\u00a0mg\u00a0g\u22121, and K+ 0.60\u00a0mg\u00a0g\u22121. Flax seeds were applied to the selected endophytic bacterial inoculum (106\u00a0CFU/ml) using carboxymethyl cellulose (CMC) solution (5%) in the range of 500\u00a0g seeds/125\u00a0ml of inoculum mixed with 25\u00a0ml of CMC. Regarding co-inoculation application, both bacterial strains were counted separately, then added to the soil. Uniform seeds were selected to dry in shade before planting  of endophytic bacterial strains  and their co-inoculation. Pots were set as follows: i- Saline soil control, ii- saline soil\u2009+\u2009A. brasilense, iii- saline soil\u2009+\u2009P. geniculata, and iv- saline soil\u2009+\u2009mixture of A. brasilense and P. geniculata. After 9\u00a0weeks of planting, plant samples were collected for morphological and physiological parameters.A pot study was undertaken in the greenhouse of the Desert Research Center's microbiological unit in Cairo, Egypt. Under normal conditions such as temperature, humidity, and light. Flax (Lengths (cm) of shoots and roots, fresh weights (g) of shoots and roots, dry weights (g) of shoots and roots, as well as number of leaves of flax plants were manually estimated using a ruler and a scale.The chlorophyll and carotenoids contents of fresh leaves of flax plants were determined using a method described by Vernon and Seely  and LichThe unit mg/g fresh weight (FW) was used to express about these contents.A described method of Umbreit et al.  was applTo assay, the soluble proteins in the dried tissues of flax plants, the method of Lowry et al.  was carrThe procedure of Bates et al.  was follTo determine the contents of ascorbic acid in the dry shoots of flax plants, half a gram was ground with liquid nitrogen and suspended in 2\u00a0ml of trichloroacetic acid (5%) and then exposed to cool centrifugation 15\u00a0min at 10,000\u00a0rpm. The supernatant (2\u00a0ml) was mixed with 8\u00a0ml of trichloroacetic acid (10%). The mixture was shaken and kept in an ice bath for 5\u00a0min and centrifuged for another 5\u00a0min at 3000\u00a0rpm. Finally, 5\u00a0ml of the extract was reacted with 2\u00a0ml of distilled water and 2\u00a0ml of Folin\u2019s reagent. After 10\u00a0min, the appeared blue color was measured spectrophotometrically at 760\u00a0nm (Jagota and Dani 2CO3 solution (1\u00a0ml) then distilled water (3\u00a0ml) were added and homogenized well. After one hour, the developed blue color was measured using a spectrophotometer at 725\u00a0nm.The phenolic content value was observed in dried shoots of flax plants using the procedure described by Dai et al.  as the fThe described steps in the method of Heath and Packer (1968) were followed for the determination of malondialdehyde contents (as an indication of lipid peroxidation) in fresh leaves of flax plants. Samples were homogenized with trichloroacetic acid (5%) and centrifugated for 10\u00a0min at 4000\u00a0rpm. After filtration, 2\u00a0ml of the extract was reacted with 2\u00a0ml of Thiobarbituric acid (0.6%) solution, then the mixture was placed in a water bath for 10\u00a0min until cooled. Read the developed color at 532, 600, and, subsequently, 450\u00a0nm.2O2) in fresh leaves of flax plants were estimated according to the described procedure in Mukherjee and Choudhuri (2SO4 (20%) (v:v). This mixture was centrifuged for 15\u00a0min at 6000\u00a0rpm. The developed yellow color of the supernatant was spectrophotometrically measured at 415\u00a0nm.The contents of hydrogen peroxide (Hhoudhuri  method. +) and potassium (K+) contents of the digested samples using flame photometry.Dry flax shoot samples (0.1\u00a0g) had been digested for 12\u00a0h with a solution of perchloric acid (80%) and concentrated sulfuric acid (1:5). The described technique of Williams and Twine (1960) was used to measure sodium , Amin et al.  and AbdeSuperoxide dismutase (E.C.1.15.1.1) activity was determined by measuring the repression of pyrogallol auto-oxidation as described in Marklund and Marklund . The sol2O2 after addition of 2\u00a0ml of 20\u00a0mM pyrogallol, the rate of increase in absorbance as pyrogallol was determined spectrophotometrically by UV\u2013visible spectrophotometer within 60\u00a0s at 470\u00a0nm and 25\u00a0\u00b0C (Bergmeyer Peroxidase (E.C.1.11.1.7)\u00a0activity was assayed using solution containing 5.8\u00a0ml of 50\u00a0mM phosphate buffer pH 7.0, 0.2\u00a0ml of the enzyme extract and 2\u00a0ml of 20\u00a0mM H2O2. The changes in absorbance were spectrophotometrically read at 290\u00a0nm (Chen and Asada For determination of ascorbate peroxidase (E.C.1.11.1.11)\u00a0activity, 0.2\u00a0ml of enzyme extract mixed with 0.5\u00a0mM of ascorbic acid, 0.8\u00a0ml of potassium phosphate buffer , 0.1\u00a0mM HStatistical analysis was performed by one-way variance analysis (ANOVA) to assess a relevant difference between different treatments using CoStat . The least significant difference (LSD) at P\u2009\u2264\u20090.05 has been used to imply a significant difference among treatments. Principal components analysis (PCA) was conducted using Minitab (version 19.0).Azospirillum spp. and Pseudomonas spp. in the rhizosphere region compared to uninoculated ones. The same patterns were detected concerning CO2 evolution as an indicator of microbial activity in the rhizosphere, where the maximum increase was measured in plants inoculated with a mixture of bacterial inoculum (138.5%).Results in Table A. brasilense and P. geniculata) showed different responses in their growth characteristics . Furthermore, co-inoculation bacterial species A. brasilense and P. geniculata markedly enhanced the growth criteria of salinity-stressed flax plants represented in shoot length, root length, fresh weight of shoot, dry weight of shoot, fresh weight of root, dry weight of root, and number of leaves about 47.9%, 30%, 19.5%, 14.9%, 69.2%, 66.1%, and 40.9% respectively.Under salinity stress conditions, inoculation of flax plants with endophytic bacterial isolates . Flax plants that were grown under salinity stress conditions and inoculated with the bacterial isolate A. brasilense observed significant increases in photosynthetic pigments, e.g., Chl. a, Chl. b, Chl. a\u2009+\u2009b and carotenoids by 9.5%, 48%, 24.4%, 84.6% respectively, when compared to un-inoculated and salinized control plants. Moreover, treating stressed flax plants with endophytic bacterial strain P. geniculata displayed significant enhancements in Chl. a, Chl. b, Chl. a\u2009+\u2009b and carotenoids by 6.47%, 42.9%, 23.29% and 61.54%, respectively. Dual inoculation with the two endophytic bacteria (A. brasilense and P. geniculata) significantly increased the contents of Chl. a (1.36%), Chl. b (20.47%), Chl. a\u2009+\u2009b (10.23%) and carotenoids (55.38%).The observed results in Fig.\u00a0A. brasilense by 69.5%, 73.1%, and 17.46%, respectively. Pseudomonas geniculata inoculation insignificantly increased sugar contents about 34.6% and significantly increased protein and proline contents by 135.35% and 7.94%, respectively, in salinity-stressed flax plants when compared with the un-inoculated plants. Co-inoculation with two bacterial strains (A. brasilense and P. geniculata) insignificantly increased the contents of sugars by 13.19% while significantly increased the contents of soluble protein and free proline by 48.60% and 1.58% respectively, in shoots of stressed-flax plants.The response of salinity-stressed flax plants to inoculation with endophytic bacterial isolates was displayed in Fig.\u00a0A. brasilense and P. geniculata) inoculation on total phenols and ascorbic acid content in flax plants grown in salinized soil was demonstrated in Fig.\u00a0A. brasilense significantly increased the contents of total phenols by 50% and ascorbic acid by 15.38% in flax plants grown under salinity stress conditions. Moreover, salinity-stressed flax plants that were inoculated with the bacterial strain P. geniculata observed a significant increase in the contents of total phenols by 64.28% and ascorbic acid by 40.67% when compared to un-inoculated stressed plants. Co-inoculation with two bacterial strains A. brasilense and P. geniculata diminished salinity stress effects in flax plants by raising total phenols and ascorbic acid contents by 28.57% and 25.12% respectively.The impact of endophytic bacterial isolates (A. brasilense and P. geniculata) inoculation on the contents of malondialdehyde and hydrogen peroxide in flax plants grown in salinized soil was shown in Fig.\u00a0A. brasilense significantly decreased the contents of MDA by 40.77% and H2O2 by 15.85% in flax plants grown under salinity stress conditions. Moreover, salinity-stressed flax plants that were inoculated with the bacterial strain P. geniculata observed significant decreases in the contents of MDA by 42.52% and H2O2 by 15.03% when compared to un-inoculated stressed plants. Dual inoculation with two bacterial strains A. brasilense and P. geniculata diminished salinity stress effects in flax plants by declining MDA and H2O2 contents by 23.55% and 13.59%, respectively.Effect of endophytic bacterial isolates  content by 42.74%, 52.75%, and 46.53%, respectively (Table +) content was significantly increased\u00a0by 36.67%, 63%, and 43.57, respectively . Also, endophytic bacterial strain P. geniculata significantly increased antioxidant enzymes activities in flax plants such as SOD, POD, and APX by 125%, 77.77% and 49.26%, respectively. Co-inoculation with the two bacterial strains A. brasilense and P. geniculata showed insignificant enhancements in the activity of SOD by 8.33%, POD by 21.29% and APX by 18.38%.Activities of antioxidant enzymes in salinity-stressed flax plants that were inoculated with the endophytes 2O2 and sodium (Na+) content. The score plot  was performed to determine the maximum amount of data variability and to study the interaction between variables and endophytic bacterial strains application. treatments. The two principal components PC1 (73.50%) and PC2 (20.30%) explained a total of 93.80% overall data variability. Loading plots of various variables Fig.\u00a0A indicatlot Fig.\u00a0B indicatAzospirillum lipoferum isolates decreased the harmful effect of salinized soil stress. As the inoculation enhanced the dry weights, number and dry weight of branches, total chlorophyll, sodium, and potassium content of wheat plants  as a means of mitigating the negative effects of salinity stress. Our findings are consistent with those of Mazhar et al.  and canola (Brassica napus)  which support plants against salinity stress throughout enhancing the antioxidant defense system  and increasing the uptake of potassium content (K+). This beneficial role may be related to the exploration that endophytes facilitates the corporation of crucial elements in the soil to be rapidly consumed by the plant  and ascorbate peroxidases (about 41%). In another study, the activity of ascorbate peroxidase was increased in stressed basil (Ocimum basilicum L.) plants in response to the inoculation with endophytic bacterial species Pseudomonas sp. and A. brasilense either individually or in combination  were highly and positively linked with two another and negatively with MDA, H2O2 and sodium (Na+) content. Score plots grouped showed the best bacterial effect on flax plants. This figure suggested that plant growth-promoting rhizobacterial strain P. geniculata was the best treatment in modulating all parameter studies as this was found in a right rectangle of the first component, whereas the Co-inoculation of A. brasilense and P. geniculata were found on the left-hand side of the first component.To validate our results on the evaluation of bacterial strains in flax plants, the entire data were subjected to a PCA-based clustering method Fig.\u00a0A, B. FigAzospirillum brasilense and Pseudomonas geniculata) play an important role in coping with salinity-induced stress throughout promoting morphological parameters, chlorophylls and carotenoids pigments, soluble sugars, proteins, free proline, total phenols, ascorbic acid, and potassium contents. Application of endophytes reduced the oxidative stress by enhancing the activities of antioxidant enzymes  in addition to minimizing the contents of malondialdehyde, hydrogen peroxide, and sodium. So, utilization of Azospirillum brasilense and Pseudomonas geniculata can be recommended as a promising method in salinity stress alleviation in plants, especially the isolate Pseudomonas geniculata which observed significant improvements in most parameters.From the observed results in the present study, it can be concluded that the tested endophytic bacteria ("}
+{"text": "Antiphospholipid syndrome (APS) with isolated cortical vein thrombosis (ICoVT) is an extremely rare but potentially malignant entity. It is particularly challenging to diagnose APS-related ICoVT because of the non-specific clinical manifestations and the frequent absence of typical neuroimaging. Moreover, there is currently limited knowledge on the clinical features and management strategies for the condition. Delays in diagnosis and treatment may lead to life-threatening consequences.We present a rare case of a 74-year-old Chinese woman who presented with sudden onset of headache and right arm weakness that mimicked acute ischemic stroke. Her initial computed tomography was unremarkable, and intravenous thrombolysis was performed. Serial neuroimages confirmed ICoVT 4 days after symptom onset, and low-molecular-weight heparin (LMWH) was started at a dose of 0.4 ml twice per day, according to the 2019 Chinese guidelines. The workup for the predisposing causes of ICoVT revealed triple positivity APS. LMWH dose was adjusted according to the anti-Xa chromogenic assay. However, the patient\u2019s condition deteriorated rapidly, and there was a progressive enlargement of the venous infarction despite treatment with anticoagulants. Transtentorial herniation developed on day 12, and decompressive craniectomy was immediately performed. The patient\u2019s symptoms did not improve significantly after surgery, and she remained aphasic and hemiplegic at the 3-month follow-up, with a modified Rankin Scale score of 5.ICoVT is a rare yet potentially fatal manifestation of APS, and its diagnosis and treatment are extremely challenging. Timely diagnosis, prompt treatment, and close monitoring are essential to improve the clinical prognosis of patients with APS-related ICoVT. Isolated cortical vein thrombosis (ICoVT) is a relatively rare and poorly understood phenomenon that represents <0.1% of all stroke entities and 6%\u20138.7% of all cerebral venous sinus thrombosis (CVST) cases . ICoVT iAntiphospholipid syndrome (APS) is a non-inflammatory autoimmune disease characterized by persistent circulating antiphospholipid (aPL) antibodies , 7. CommHere, we present an extremely rare case of a primary APS patient who presented with malignant ICoVT as an initial presentation, whose condition rapidly deteriorated despite anticoagulation treatment. This case highlights our concern among clinicians for this rare yet potentially fatal disease. We discuss the lessons that can be learned from the diagnosis and management of this rare disease and how such lessons can aid in improving patient care and outcomes. Furthermore, we discuss the importance of radiologic monitoring and anti-Xa activity measurement to guide anticoagulant therapy for patients experiencing clinical deterioration.A 74-year-old Chinese woman (gravida 3 para 3) was admitted to the emergency department with sudden onset of headache and right arm weakness for 2.5 h. She had a history of well-controlled hypertension and hyperlipemia, for which she took 5 mg of amlodipine daily and 10 mg of atorvastatin daily as instructed by her primary care provider. She had no family history of thrombotic disease and no history of venous thrombosis. She also denied any history of risk factors for thrombosis .On admission, the patient\u2019s vital signs were within normal limits except for a slightly elevated blood pressure of 141/89 mmHg. Her weight was 63 kg. Neurological examination showed Medical Research Council (MRC) grade 3 for the right arm. The National Institute of Health stroke scale (NIHSS) score at admission was 2. Non-contrast computed tomography (NCCT) was performed, and the radiologist reported no acute abnormalities . Her D-dimer level was 4.87 \u00b5g/ml. Other laboratory workups were negative and included blood routine tests, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), liver and renal function, lipids panel, glycated hemoglobin, homocysteine, infection panel, and tumor markers . A repeat CT at 24 h showed no hemorrhage . However, diffusion restrictions, which are the key parameters for identifying cerebral infarctions, were absent in the parenchyma on diffusion-weighted imaging. After consultation with radiologists, a convexity subarachnoid hemorrhage (cSAH) was considered. Given that the patient was clinically stable, aspirin was continued. CT angiography was performed on day 4 and did not find any aneurysm or arteriopathy. Aneurysmal subarachnoid hemorrhage (SAH) was excluded. However, filling defects of CoVs and compensatory expansion of the adjacent veins were present  and fluid-attenuated inversion recovery imaging (FLAIR)] that neighbored the cerebral falx and left frontal sulcus (The patient complained of severe headache and progression of right limb weakness (MRC grade 3/5). The MRI on the night of day 4 showed a venous infarction in the frontal lobe , anticardiolipin (aCL)], and anti-\u03b22-glycoprotein-I (a\u03b22GPI) antibodies] was performed on the morning of day 5 before the patient underwent anticoagulation. Additionally, screening for an underlying malignancy was initiated, including chest CT, transthoracic echocardiography (TTE), abdominal ultrasound, and transvaginal ultrasound. However, no evidence of malignancy was found. In the absence of infective symptoms, systemic infection  was excluded according to normal white blood cell count, urine analysis, CRP, ESR, procalcitonin TTE, and chest CT. Laboratory workup reported the next day were positive for LA , aCL IgG , aCL IgA , a\u03b22GPI IgG , a\u03b22GPI IgA . Factor VIII, Factor VII, and von Willebrand factors significantly increased by 225% , 212% , and 160% , respectively.2O, and cerebrospinal fluid analyses were all within the normal range. Thus, intracranial infection, SAH, and intracranial hypertension were subsequently excluded. LMWH was restarted 4 h after the lumbar puncture.Aspirin was stopped on the morning of day 5, and low-molecular-weight heparin (LMWH) was started at a dose of 0.4 ml every 12 h according to the 2019 Chinese guidelines for clinical management of CVST . On the MR black-blood-thrombus imaging (MRBTI) on day 7 showed vein thrombosis that extended proximally to the superior sagittal sinus  in the left frontoparietal lobe and transtentorial herniation   . In addiSeveral therapeutic options are available for APS-CVST patients who are refractory to anticoagulant therapy. Current evidence does not support the use of systemic thrombolysis before anticoagulant therapy. Ongoing anticoagulant therapy is a contraindication of systemic thrombolysis. Recent years have seen an increase in the use of endovascular therapy (EVT), including intrasinus thrombolysis (IST) and mechanical thrombectomy (MT), in patients who are refractory to anticoagulant therapy . ThromboDecompressive craniectomy (DC) is a well-established life-saving procedure and also the only treatment for malignant CVST or ICoVT . BecauseICoVT is a rare yet potentially fatal manifestation of APS. Clinicians should consider ICoVT in patients who present with an increased D-dimer level and symptoms of focal neurologic deficit and/or seizures. APS should be considered in patients with ICoVT, particularly young women. Neuroimaging is the cornerstone of ICoVT diagnosis, and multi-parametric MRI is required to identify this rare entity. Prompt anticoagulation and close monitoring are essential to achieve good clinical outcomes in patients with APS-related ICoVT.The original contributions presented in the study are included in the article. Deidentified data, including clinical manifestations, neuroimaging data, serum tests, and cerebrospinal fluid tests, are available upon appropriate request to the corresponding author.Written informed consent was obtained from the individual(s) for the publication of any potentially identifiable images or data included in this article.FF and YL contributed to the design of the study, revised the manuscript, and were responsible for the integrity and accuracy of the data. JShen contributed to drafting the manuscript and reviewing the published literature. ZT contributed to the collection of clinical and neuroimaging data. JSun was the attending doctor of the patient and contributed to the acquisition and analysis of the clinical data. WC contributed to the analysis of the neuroimaging data. All authors contributed to the article and approved the submitted version.This work was supported by the Medical Science and Technology Project of Zhejiang Province of China (No. 2021KY797), the Natural Science Foundation of Zhejiang Province of China (No. LY21H090015), and the Wenzhou Basic Scientific Research Project (No. Y20210900).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."}
+{"text": "Multi-modal magnetic resonance imaging (MRI) is widely used for diagnosing brain disease in clinical practice. However, the high-dimensionality of MRI images is challenging when training a convolution neural network. In addition, utilizing multiple MRI modalities jointly is even more challenging. We developed a method using decomposition-based correlation learning (DCL). To overcome the above challenges, we used a strategy to capture the complex relationship between structural MRI and functional MRI data. Under the guidance of matrix decomposition, DCL takes into account the spike magnitude of leading eigenvalues, the number of samples, and the dimensionality of the matrix. A canonical correlation analysis (CCA) was used to analyze the correlation and construct matrices. We evaluated DCL in the classification of multiple neuropsychiatric disorders listed in the Consortium for Neuropsychiatric Phenomics (CNP) dataset. In experiments, our method had a higher accuracy than several existing methods. Moreover, we found interesting feature connections from brain matrices based on DCL that can differentiate disease and normal cases and different subtypes of the disease. Furthermore, we extended experiments on a large sample size dataset and a small sample size dataset, compared with several other well-established methods that were designed for the multi neuropsychiatric disorder classification; our proposed method achieved state-of-the-art performance on all three datasets. Many neuropsychiatric disorders (NDs) not only result in a huge socioeconomic burden but are also accompanied by several comorbidities  is a useful tool for clinical diagnosis of ND. It can provide information on different aspects of the brain. Functional MRI (fMRI) can be used to analyze the functional connections (FCs) between different brain regions. These FCs reveal individual differences in neural activity patterns, which can predict continuous phenotypic measurements  and a distinctive matrix (Dk), respectively. Based on the output  = 0, and cbase \u2208 .where the constraints Ck can be defined as follows:Then, the estimator of C1 and C2 have the maximum correlation between each other, while the vectors within each are uncorrelated and whitened. Their correlation vectors where Dk is defined as follows:The estimator of Xk, is defined as follows:In our experiment, we use the relationship between We summarize DCL in We evaluated the proposed DCL method in classifying NDs in the CNP dataset , 50 SZ subjects, 49 BD subjects, and 43 ADHD subjects. These 272 images were from people in the Los Angeles area aged between 21 and 50 years old who were recruited through community advertisements. The details of the CNP dataset are listed in Brain connectivity information may be reflected in fMRI images. In the CNP dataset, each sample has seven fMRI modalities, which were collected during different task states: BOLD contrast, resting state , breath-holding tasks , balloon analog risk tasks, stop-signal tasks, task switching, and spatial working memory capacity tasks. In this study, we attempted to classify NDs using resting-state fMRI images.Resting-state fMRI is an imaging technique that obtains a brain activity function map when the subject is in a resting state undisturbed by other activities, which is better for distinguishing ND groups. The CNP dataset has resting-state fMRI images with scans lasting 304 s. The participants were relaxed with their eyes open. They were not stimulated or asked to respond during scanning  classifying NDs into different subtypes using fMRI and sMRI data and (2) extracting important features from the fMRI and sMRI images. The classification task was to validate the performance of the DCL method for the different ND groups, whereas the feature extraction task was used to assess the capability of the method in detecting correlated features.n samples, verification based on LOO is carried out over n iterations. In each iteration, the classifier uses n \u2212 1 samples as training samples and uses the remaining sample as testing samples.We obtained the correlation matrices by inputting the 248 fMRI (90 \u00d7 90) and sMRI (248 \u00d7 2196) matrices into S-POET. Then, we decomposed each pair of canonical matrices and computed their correlations. Finally, we used the leave-one-out (LOO) method to select the important features in the test sample matrix. For a dataset with F1) are used to measure the classification performance. They are defined as follows:In our experiments, accuracy (ACC), precision (PRE), recall (REC), and F-score , XGBoost, PCA+SVM, PCA+RF, PCA+XGBoost, CCA+SVM, CCA+RF, and CCA+XGBoost. The linear kernel in the SVM classifier was used, as it provides better experimental performance than other kernels. As a trade-off between performance and computational cost, we set the number of trees in RF to 100. To prevent overfitting by XGBoost, we set the maximum tree depth for base learners and the turning parameter for the The classification results for the DCL method and the other classifiers are shown in In addition, our comparative experiment was based on a sample size of 248. As shown in Machine learning methods can be effective for classifying simple images, but because medical images are very complex, these three machine learning methods were overwhelmed.The limited sample size does not meet the training requirements of the three machine learning methods. The multi-class classification task increased the imbalance for the samples, making it difficult for these methods to obtain key feature information from the high latitude and limited samples.Therefore, unlike the other methods, the DCL method first preprocesses the complex relationship between the sMRI and fMRI data, which reduces the complexity of the input data. Of the DCL-based classifiers, XGBoost had the best results in the multi-class classification task. The best accuracy was 72.00%. The receiver operating characteristic (ROC) curves for XGBoost in multi-class classification is plotted in Besides assessing the performance of the DCL method, we also identified the important features with the DCL+XGBoost method. The aim was to find which edges contribute to brain connectivity. The extracted features are mapped back into the brain space, which facilitates the interpretation of the known relationship between brain structure and function. However, due to the dimensionality of the connectivity network, the visualization is challenging. In the LOO method, we used a weight-based method to evaluate the importance of features in the test sample matrix. The weight in XGBoost is used to calculate the number of times a feature is used as a split point across all trees. Finally, we counted the number of samples whose feature weights were >0. We visualized the representations of all important features for both the sMRI and fMRI data.https://www.nitrc.org/projects/bnv/) to visualize which FCs have the strongest relationships in the brain network.It is interesting to investigate how different brain networks cooperate and connect with each other. We found that there were significant differences between the FCs of each group, which indicates that these FCs not only reflect the information common to the different groups but also the differences among them. We used the BrainnetViewer software  and a small sample size dataset (a subset of OpenfMRI), respectively.The shrinkage principal orthogonal complement thresholding method is a covariance estimator with the approximate factor model, which is based on sparse PCA. In our method, we used the S-POET method to obtain asymptotic first-order distribution for the eigenvalues and eigenvectors of the fMRI and sMRI correlation matrices, respectively. To verify the effect of the S-POET method in our proposed DCL method, we extended two different DCL methods on XGBoost: one is based on PCA[DCL(PCA)] and another is based on S-POET[DCL(S-POET)].As shown in To verify the influence of different MRI modalities on model, we separately used fMRI, sMRI, and fMRI+sMRI matrices as inputs to three types of XGBoosts, namely PCAXGBoost, CCA-XGBoost, and DCL-XGBoost.The results are shown in Some patients in the ND group had taken medication for their mental illness. To analyze the impact of these medications on the patients, we visualized the selected FCs for a group who had taken medication and for a group who had not. There are significant differences between these two groups, as shown in To verify the performance of DCL on different datasets, we extended experiments on a larger sample size dataset (a subset of ADNI) and a small sample size dataset (a subset of OpenfMRI), respectively. The Alzheimer's Disease Neuroimaging Initiative (ADNI)  We used only MRI data as the input. However, the classification of complex disorders could be made more accurate by including phenotypic information. (2) The amount and uneven quality of the MRI data have a significant influence on the performance of a model and reduce the accuracy of classification.This work demonstrated that the DCL method can effectively combine different information from fMRI and sMRI images. DCL identifies both the common and distinct information between the two input MRI matrices. The decomposition-based CCA is used to analyze the correlation and construct the required matrices. Thus, DCL has better performance in both classification and identifying FCs. The DCL method can be used to detect complex and nonlinear relationships between the two types of MRI images. Our experiments showed that the DCL method can improve classification performance so that it is a suitable method for classifying mental illnesses.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s.LL, HZ, and GL contributed to the conception and design of the study. JC organized the database. YW performed the statistical analysis. LL wrote the first draft of the manuscript. Y-PW modified the manuscript. All authors contributed to manuscript revision, read, and approved the submitted version.This work was funded partially by the National Natural Science Foundation of China under Grant Nos. 62172444, 61877059, and 62102454, the 111 Project (No. B18059), and the Henan Provincial Key Research and Promotion Projects (No. 222102310085).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."}
+{"text": "Results revealed a large genetic variation in mostly morphological traits under low P. Significant enhancement in root traits and phosphorus efficiency-related traits like PUE was observed at LP as compared to CK conditions. In response to low P availability, cotton genotypes showed large plasticity in shoot and total dry biomass, phosphorus, and nitrogen efficiency-related traits , and most root traits, but a limited response in root dry biomass, taproot length, root surface area, root volume, and SPAD value. In addition, significant correlations were observed between PUtE (phosphorus uptake efficiency), NUE (nitrogen use efficiency), TDB , and RTD (root tissue density) with PUE under both P supply level and phosphorus stress index, which may be a key indicator for improving PUE under LP conditions. Most root traits are most affected by genotypes than nutrition level. Conserved PUE is more affected by the nutrition level than the genotype effect. Principal component analysis depicted the comprehensive indicators under two P supply conditions were mainly reflected in root-related traits and morphological indicators such as dry matter biomass. These results indicate that interspecific variations exist within these cotton genotypes and traits. Our study provides suggestions for future research to enhance the ability of the earth system model to predict how crops respond to environmental interference and provide target quality for cotton breeding in phosphorus-deficient areas.Phosphorus (P) is an essential macronutrient required for fundamental processes in plants. Trait plasticity is crucial for plant adaptation to environmental change. Variations in traits underlie diverse phosphorus (P) acquisition strategies among plants. Nevertheless, how the intraspecific plasticity and integration of morphological traits contribute to Phosphorus-Use-Efficiency (PUE) in cotton is unknown. In this study, 25 morphological traits were evaluated in 384 cotton genotypes grown with low P (LP, 10\u03bcmol. L As a component of vital macromolecules including nucleic acids, nucleotides, and phospholipids 43 as well as having major metabolic and protein-regulating functions in the form of phosphate (Pi) or Pi 44 esters, phosphorus (P) is one of the most important essential nutrients . Plants Plants have various adaptive strategies for boosting P absorption and mobilization to deal with reduced P availability in soil . Plants Mainly the P acquisition features have been extensively utilized to increase the PUE of crops . But it\u2019Cotton (Gossypium hirsutum L.) is a vital crop for the global economy that produces natural fiber, the primary raw material for the textile industry . Cotton The research was set up in the growth chamber at the Institute of Cotton Research, the Chinese Academy of Agricultural Sciences, Anyang, China. The 384 cotton genotypes used in this experiment are based on a breed population determined in previous studies (not published). The breeding years information is shown in 6Mo7O24\u00b74H2O, 0.5mmol/L KH2PO4, 2.5mmol/L Ca (NO3)2\u00b74H2O, pH 5.5) containing 10 (LP), 500 (CK) \u03bcM KH2PO4. Every week new solutions were added, and an electric pump aerated them. Plants started to show clear signs of P treatment after four weeks. Then various morphological characteristics were measured.384 cotton genotypes seeds were incubated for 7 days in a sand and vermiculite combination in a growth chamber. The uniform seedlings were transplanted into 8L plastic boxes after germination. The plants were grown under natural light in a greenhouse. 28/20\u00b0C (day/night) temperatures, 60% humidity levels. At the initial stage, the plants were provided with 100 mL dHposition . A totalThe dry biomass of the shoots and roots was then calculated after 48 hours of drying at 72\u00b0C. Roots were dipped in 0.1% (w/v) toluidine blue for 5min and then scanned at 300 dpi using a desktop scanner . Afterward, root images were loaded into a WinRHIZO analysis system . The root traits were determined according to the established method . A porta+ Luebbe Continuous-Flow AutoAnalyzer III (AA3-Germany) after 3 hours of digestion with H2SO4-H2O2. PUE and NUE definitions for cotton genotypes grown at various phosphorus concentrations are also included and were calculated as Total P and N accumulation  = the P and N concentration multiplied by total plant dry matter, P and N uptake efficiency  = the total plant dry matter divided by P and N concentration, P and N use efficiency  = TPA, TNA divided by root dry matter, follows by  and tested for homogeneity of variance (Leven median test). The Global ANOVA was used to evaluate the data (genotype and nutrition level as main factors), and the means were separated by Tukey\u2019s honest significant difference (HSD) test (P < 0.05). The standard deviation to a collection means ratio was used to estimate the coefficient of variation for each parameter.2PL) was estimated as + Residuals, where nutrition level was treated as a fixed effect, and Genotype and Genotype\u2010by\u2010nutrition level were treated as random effects. Conversely, trait broad sense heritability (h2B) was calculated as The term \u201cphenotypic plasticity\u201d (PL) refers to the alteration in phenotype brought on by environmental variance . This isPL/\u03c32\u00a0P . FinallyPrincipal component analysis (PCA) was calculated in OriginPro (2021) . Morphophysiological traits were used to calculate the correlation relationships in Microsoft excel. The figures were drawn with OriginPro (2021). All the data results are expressed as the mean of four replications. Biplot graphs for each characteristic displayed the average values for each accession obtained at LP  compared to those at CK  levels. We could determine which attribute was most significantly impacted by the P supply due to the biplot analysis. The distribution of genotypes along the diagonal bisector of biplots explained their genetic variation. The distance from the bisector revealed each genotype\u2019s ability to adapt to the P availability.Cotton genotypes\u2019 responses to the two P treatments were significantly different in morphological characteristics  of 0.32 and 0.36, respectively, compared to CK conditions. A similar trend was observed for SFW and RFW  interactions, accounting for 2%\u201348% of the total variance. They played a substantial part in the total variance Figure\u00a06To find correlations between the traits, morphological traits\u2019 relative values were correlated (Pearson test). There were significant connections for most of these traits Figure\u00a072 = 0.65, P<0.001) and LP  conditions  and LP  conditions, but it was not significant  and LP  conditions  and PUtE , under the CK condition, have the same trends  was carried out. 25 features were obtained from the average of both cotton genotypes for the loading plots of PC1 and PC2 separately S5. In tIn this study, eight principal component factors were extracted from PCA analysis based on the relative values of each morphological trait Figure\u00a09In this research, there were significant differences in dry matter accumulation of cotton under LP and CK conditions. The TDB decreased by 51.95% in LP conditions compared to CK conditions . AccordiIn this study, the decreasing PUE under CK conditions may result from the accumulation of external phosphorus beyond what is required for growth, negatively impacting a plant\u2019s P economy . In this(Cicer arietinum L.) germplasm accessions exhibit a surprising degree of variety in root shape  and the Central Public-interest Scientific Institution Basal Research Fund, CAAS (Y2021XK12).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher."}
+{"text": "Mutations in the KCNH2 gene encoding hERG are associated with several cardiac rhythmic disorders, mainly the Long QT syndrome (LQTS) characterized by prolonged ventricular repolarization, leading to ventricular tachyarrhythmias, sometimes progressing to ventricular fibrillation and sudden death. Over the past few years, the emergence of next-generation sequencing has revealed an increasing number of genetic variants including KCNH2 variants. However, the potential pathogenicity of the majority of the variants remains unknown, thus classifying them as variants of uncertain significance or VUS. With diseases such as LQTS being associated with sudden death, identifying patients at risk by determining the variant pathogenicity, is crucial. The purpose of this review is to describe, on the basis of an exhaustive examination of the 1322 missense variants, the nature of the functional assays undertaken so far and their limitations. A detailed analysis of 38 hERG missense variants identified in Long QT French patients and studied in electrophysiology also underlies the incomplete characterization of the biophysical properties for each variant. These analyses lead to two conclusions: first, the function of many hERG variants has never been looked at and, second, the functional studies done so far are excessively heterogeneous regarding the stimulation protocols, cellular models, experimental temperatures, homozygous and/or the heterozygous condition under study, a context that may lead to conflicting conclusions. The state of the literature emphasizes how necessary and important it is to perform an exhaustive functional characterization of hERG variants and to standardize this effort for meaningful comparison among variants. The review ends with suggestions to create a unique homogeneous protocol that could be shared and adopted among scientists and that would facilitate cardiologists and geneticists in patient counseling and management.hERG, the pore-forming subunit of the rapid component of the delayed rectifier K KCNH2 gene encoding hERG channel), allows identifying the relatives at risk, in the proband\u2019s family  is a cardiac disorder characterized by abnormally prolonged ventricular repolarization that results in episodic ventricular tachyarrhythmias sometimes leading to ventricular fibrillation and sudden death in otherwise healthy persons . LQTS iss family  , the Association for Molecular Pathology (AMP) and the College of American Pathologists (CAP) proposed guidelines that compiles all the parameters that contribute to predict the potential pathogenicity of a given variant, namely the patient phenotype, the segregation data, the conservation of the varying sequence across species, the population data, the ds, etc. . The finhttp://gnomad.broadinstitute.org/) (The \u201cuncertain significance\u201d (VUS) usually occurs in two situations: when there are conflicting results from the different observables, or when there is insufficient evidence, as is often the case for novel missense variants. Intuitively, it seems reasonable that functional data will be significantly helpful to decrease the number of cases of \u201cuncertain significance.\u201d A theoretical approach proposed by Brnich and collaborators used an algorithm to calculate all combinations of evidence. Using this systematic counting approach, they estimate how functional data allows decreasing the number of VUS. They convincingly illustrate that a majority of VUS could be reclassified with the addition of solid functional data . This obte.org/)  are rarete.org/) . This obKCNH2 variants are referenced in Clinvar https://www.ncbi.nlm.nih.gov/clinvar/, of which 1832 are associated to LQTS. KCNH2 gene is associated with type 2 Long QT syndrome, but also with Atrial Fibrillation  are most often pathogenic, we focused our analyses on missense variants. Among these 1135 missense variants, 962 are classified as VUS, representing approximately 85%.At the writing of this review, 2434 illation  and shorillation . It reprillation . Figure This review aims at describing the nature of the functional assays undertaken so far and their limits, based on an exhaustive scan of these 1135 missense variants and additional 187 coming from the French network CARDIOGEN, not yet in ClinVar. We also looked at some specific details of the functional assays, in a smaller sample of 112 variants issued from the same French database for type 2 Long QT syndrome patients. This review then comes with new propositions to generate standardized protocols that will help cardiologists and geneticists for patient counseling in face of the increasing number of variants.As mentioned above, functional data being very helpful to reclassify the VUS, a functional evaluation of hundreds of variants is necessary. Also, the methods of acquisition of these functional data have to be scrupulously standardized to faithfully evaluate the pathogenicity of any variant. In this review, our aim was to establish how far we are from an extensive and standardized evaluation of hERG variants.We performed an exhaustive scan of the literature on the 1322 missense variants . We can Regarding functional studies, it is important to note that patch-clamp is the gold standard for assessing mutations affecting hERG functionality but to date, not all studies have extensively characterized all biophysical aspects. To understand this point, one should know that hERG channels present two gates (one activation gate and one inactivation gate) with specific voltage-dependence and kinetics. In the simplest case, maximal current is evaluated when both activation and inactivation gates are open. If this maximal current is lower in the variant as compared to WT hERG channel, there is a loss of function . But desThus, it is possible that a mutation alters a single parameter among all known biophysical parameters, namely the maximal current amplitude, the half-activation potential, the slope of the activation curve, the half-inactivation potential, the slope of the inactivation curve, activation and deactivation kinetics, inactivation and recovery from inactivation kinetics, and also the ion selectivity. This indicates how critical it is to study systematically all channel parameters of a given variant.On a sample of 38 variants that were studied in electrophysiology, we examined how exhaustive all these parameters were characterized. These 38 variants were studied in electrophysiology among a total of 112 missense variants  collected in the national database of long QT patients constituted by a network of French Centers of Reference on cardiac arrhythmias . These Centers of Reference belong to the French network CARDIOGEN, which has developed a common database extensively reporting clinical and genetic information from all identified French cases. In the case of these 38 missense variants we are far from an extensive study of hERG biophysical parameters  during hERG channel expression by the cultured cells, after introduction of the DNA/RNA encoding the channel and 2) during the electrophysiological characterization of the variants.Xenopus laevis oocytes, which require incubation at low temperature (12\u00b0C\u201318\u00b0C) and mammalian cells, typically incubated at 37\u00b0C. Noteworthy, incubation at room temperature rescues the impaired trafficking of many hERG channel variants, as observed by hERG protein glycosylation which is an indicator of correct cell trafficking  A non-mammalian model, X. laevis oocytes. The presence of endogenous currents, in these models, but also in X. laevis oocytes, may interfere with measurements when studying mutants which generate currents of low amplitudes. To that respect CHO cells are a good model since they expressed minimal voltage-gated currents, when compared to other cells such as HEK-293 cells ((ii) Mammalian cell lines that are easy to maintain and animal-free . Two cel93 cells . On the 93 cells . This ha(iii) Induced pluripotent stem cells developed by Shinya Yamanaka  allow thAnother source of variability is the cell model of study. Three categories of models are used for hERG variants phenotyping :(i) A noX. laevis oocytes, the aforementioned mutation (H70R) led to similar deactivation alteration, but had a lower impact (around 25% decrease in homozygous condition) on current amplitude . So far H70R has only been studied by Western blots from HEK-293 cells, but this technique showed opposing results: one study suggests that H70R hERG is trafficking-deficient , but similar hERG current levels in their corresponding iPS-CMs . AnotherNa current abnormality in the six cell lines, independently of the presence (2 Brugada patients) or absence (4 Brugada patients) of a mutation in the SCN5A gene, coding for the sodium channel NaV1.5  to determine if there is an equal expression of both alleles, which seems to be the case , and 2) In conventional patch-clamp study, equal expression of both alleles is achieved by transfecting cells with the equal amount of the two DNAs . Mimickiv channels and named \u201cdelayed repolarization\u201d is in fact an artifactual property due to insufficient voltage-clamp. This strongly suggests that, in the case of transient transfection, the protocol has to be finely tuned to prevent currents larger than 10\u00a0nA  is responsible for a major variability in the observed biophysical parameters . We alsoan 10\u00a0nA .In the same vein, in order to limit the artifactual heterogeneity of the recordings, it is also critical to design the most adapted voltage simulation protocol specific to a given channel, since voltage-dependence and kinetics drastically vary from one channel type to another. For instance, a steady-state activation protocol in which the depolarizing pulse are too short in time may lead to erroneous effect of a given variant, such as the negative shift in the activation curve in the case of the R176W variant .2. Such a great deal of data allowed us to add many numerical constraints on a kinetic model and to demonstrate that PIP2 stabilizes hERG channel open state  generating enough information on the current characteristics, to use it to constrain a kinetic model that will be accurate enough to describe the properties of the activation and inactivation gates. Impressively, the protocol is able to predict the current obtained by the classical protocols . A majorIn Silico pathogenicity prediction softwares, which are evaluating the evolutionary conservation of an amino acid or nucleotide, the location and context within the protein sequence, and the biochemical consequence of the amino acid substitution  and not notyping .In Silico methods will logically bring in parallel a standardized approach to predict the functional impact of a given variant.At last, development of"}
+{"text": "The gradual impact of the Covid-19 pandemic had important effects on routines in surgical environments. In order to cope with the impact and re-establish anaesthesiology and surgery procedures, it was imperative to pursue in-depth studies with a view to ensuring safe surgical care, reducing hazards, as well as protecting the health, safety and wellbeing of the health personnel involved. The purpose of this study was to evaluate quantitative and qualitative approaches to domains of safety climate among multi-professional staffs of surgical centres during the Covid-19 pandemic and to identify intersections.This mixed-method project employed a concomitant triangulation strategy on a quantitative approach in an exploratory, descriptive, cross-sectional study, as well as a qualitative approach by way of a descriptive study. Data were collected using the validated, self-applicable Safety Attitudes Questionnaire/Operating Room (SAQ/OR) questionnaire and a semi-structured interview script. The 144 participants were the surgical, anaesthesiology, nursing and support teams working in the surgical centre during the Covid-19 pandemic.The study found an overall safety climate score of 61.94, the highest-scoring domain being \u2018Communication in the surgical environment\u2019 (77.91) and the lowest, \u2018Perception of professional performance\u2019 (23.60). On integrating the results, a difference was found between the domains \u2018Communication in the surgical environment\u2019 and \u2018Working conditions\u2019. However, there was intersection by the \u2018Perception of professional performance\u2019 domain, which permeated important categories of the qualitative analysis.For care practice, it is hoped to encourage improved patient safety, educational interventions to strengthen the patient safety climate and promote in-job wellbeing on the job for health personnel working in surgical centres. It is suggested that further studies explore the subject in greater depth among several surgical centres with mixed methods, so as to permit future comparisons and to monitor the evolving maturity of safety climate. Safety climate relates to health care personnel\u2019s perceptions of how the organisation deals with safety issues. For that reason, measuring safety climate offers the opportunity to identify the perceptions and attitudes of multi-professional teams regarding patient safety issues in the surgical environment .As safety climate is intrinsically related to health service quality , managerThe Covid-19 pandemic gradually affected routines in health organisations and surgical environments making it important to assess the safety climate in surgical centres during the pandemic period with a view to planning to address the impact and re-establish anaesthesiology and surgery procedures . Given tEvidence of the pandemic\u2019s impact on safety climate includes postponed surgeries, increased complications for lack of surgical procedures, the risk of Covid-19 transmission during procedures, posing a need for new safety and flow protocols, awareness building as to appropriate, quality, rational use of personal protection equipment, stronger and more effective communication with testing laboratories and routine pre-operatory assessment .Knowing and evaluating safety climate in the perioperatory environment is expected to yield data for a situational diagnosis, continued professional development measures, introduction of care protocols and monitoring of surgical incidents . FurtherData from research into safety climate and culture help guide planning and implementation of measures with a view to creating a satisfactory and propitious working environment, with motivated staff and particularly quality of care and guaranteed safety for patients and surgical team .There is as yet little scientific evidence of safety climate studies that apply an instrument appropriate to the surgical centre setting, making it impossible to compare by the psychometric properties of its domains , 9. AccoIn view of the overall adversity resulting from the Covid-19 pandemic, it was decided to resort to a mixed method, which offers the advantage of leveraging the power of sophisticated integration of data from quantitative and qualitative approaches and, in that way, overcoming a series of challenges in the health sciences ; in thisIn view of the foregoing, the study hypothesis was that there are intersections between the quantitative and qualitative approaches to evaluating safety climate of a multi-professional surgical centre team during the Covid-19 pandemic. Accordingly, this study examined the quantitative and qualitative approaches of safety climate domains as applied to the multi-professional team of a surgical centre during the Covid-19 pandemic, so as to point up the intersections.This mixed-method study applied a concomitant triangulation strategy which made it possible to identify assimilations and divergences among the data. The quantitative portion was cross-sectional, exploratory and descriptive, while the qualitative study was descriptive. Data for the two portions were collected simultaneously.In a more practical context, the mixed method affords a more robust research approach and makes for more in-depth and complete understanding of the problems in health issues, especially in promoting safe care , 15. It The supports used were: for the quantitative research, Strengthening the Reporting of Observational Studies in Epidemiology (STROBE\u00ae) ; for theThe research setting was a surgical centre at a university hospital of Brazil\u2019s public Unified Health System  in Rio de Janeiro. This teaching, research and extension facility performs neurosurgery, surgery of the head and neck, ear, nose and throat and in urology, gynaecology, proctology, orthopaedics, vascular and plastic surgery, conventional general surgery and surgery by videolaparoscopy. On average, 324 surgeries are performed a month or approximately 3,888 per year.N\u2009=\u2009214), sample size was calculated for a simple random sample, to a 95% confidence level and sample loss of 5%. Sample size was thus 137 workers [The study population comprised all staff of the surgical centre. For the quantitative stage based on a population given by a multi-professional team working in the research setting during the pandemic  in the surgical environment in the Covid-19 pandemic?\u2019; and \u2018What do you think of, and how would you rate, your job performance in the Covid-19 pandemic?\u2019.After approval from the research ethics committee, data were collected in person by a researcher during all shifts from February to May 2021. The staff were invited to take part in the study and, after the aims of the study were explained to them, participants signed a declaration of free and informed consent. Only then were they offered the printed questionnaire. These were collected up and stored in envelopes coded by numerical sequence in which they were returned.After participants agreed to voice recording in their declaration of free and informed consent, interviews were carried out face-to-face and individually, in the study setting and, for convenience, in the order in which the quantitative questionnaires were returned. The interviews lasted about 15 to 20\u00a0min and were recorded digitally. Given the pandemic situation, a distance of 1.5\u00a0m was maintained between researcher and participant, and both used masks and visors throughout collection.During the study, the lead researcher worked in the study setting as a nursing technician and surgical technician. As a result, she already had a relationship with the participants, enabling her to approach them better, explain the study objectives and motivate them with regard to the study. Despite the existing relationship between researcher and setting, ethical requirements and relations with participants were respected in order to ensure data privacy, validity and reliability.The quantitative data were inserted manually into a Microsoft Excel\u00ae electronic spreadsheet and then input to R statistics software, version 4.1.0., a freeware tool used to analyse and treat statistical data.The instrument\u2019s reliability was evaluated by Cronbach alpha. Descriptive statistics were used for analysis of the numerical and categorical variables. In describing staff profiles, the numerical variables  were displayed in absolute values, means, medians, standard deviations, upper and lower limits. The categorical variables  were displayed as absolute and relative frequencies.In order to organise the qualitative data, the interviews were transcribed in full and later revised by the lead researcher so that nothing said was suppressed. This was done using the Microsoft Office Word\u00ae text editor software, with each interview identified by a number in the order in which they were transcribed and, so as to ensure participant anonymity, they were identified by professional team, viz.: ENF for the nursing team; CIR for surgery; ANEST, anaesthesiology; and AP, auxiliary personnel. The data then underwent content analysis and were interpreted by thematic category.This stage was carried out using Microsoft Office Excel\u00ae, spreadsheets, which permit data analysis. Accordingly, drawing on the theoretical underpinning and the literature findings, the data were analysed and interpreted on a broad view, so as to identify similarities and differences as compared with other studies , 22.The interpreted quantitative and qualitative data were then examined to detect intersections in the findings using the concomitant triangulation strategy to identify convergences and divergences in the data. The mixed method research approach investigates by combining or associating quantitative and qualitative data, integrating the two approaches in the same study and thus permitting the research problem to be understood better .To that end, the findings were presented in tables and charts to elucidate and help understand the information to be analysed. In this way, the scores for each domain were determined, as were the themes in what was said on each domain and the prevalence of mentions in the qualitative approach. That said, the quantitative and qualitative data were analysed separately and then the results from both were compared so that differences and similarities could be highlighted in the data intersection section.The study was approved by the research ethics committee of the proponent institution under CAAE: 40571920.1.0000.5258 and was conducted in compliance with resolutions current in Brazil. All data were preserved and analysed confidentially and scientifically.The highest-scoring patient safety climate domain was \u2018Communication in the hospital environment\u2019. The study findings demonstrated the reliability of the instrument, which returned an overall Cronbach alpha score of 0.717. The various domains scored as follows: \u201cSafety climate\u201d, 0.784; \u201cPerception of management\u201d, 0.669; \u201cPerception of stress\u201d, 0.757; \u201cWorking conditions\u201d, 0.708; \u201cCommunication\u201d, 0.616; and \u201cPerception of job performance\u201d, 0.770.The quantitative study participants were 144 staff of the university hospital surgical centre. Their social and demographic characteristics were as follows: 73 (50.69%) were male, 75 (52.08%) were under 39\u00a0years old. By profession, the team that predominated was medical  with 92 members (63.89%), followed by the nursing team  with 48 (33.33%) and the support team  with 4 (2.78%). Note that the professional category with most representatives was surgical technicians and circulating nurses, with 38 personnel (26.39%). The participants\u2019 characteristics are shown in Table The mean overall SAQ/OR score was 61.94 dp\u2009\u00b1\u200912.41). Note that, for a patient safety domain to be considered positive required a score of more than 75. The highest-scoring safety climate domain was \u2018Communication in the surgical environment\u2019 (77.91) and the lowest-scoring was \u2018Perception of job performance\u2019 (23.60). The safety climate domain scores for the study setting are shown in Table .41. NoteThe qualitative study participants were distributed as follows: 10 members of the nursing team, 7 of anaesthesiology, 6 of surgery and 1 of the support service. It was decided to take the terms that participants used to name units of meaning as the study categories and to discuss the most prevalent.The categories discussed were as follows: from the \u2018Working conditions\u2019 domain 38.99%), \u2018COVID-19 Protocol' (18.87%); from the \u2018Safety climate\u2019 domain (16.35%), \u2018Patient safety\u2019 (8.18%); from the \u2018Perception of stress\u2019 domain (13.84%), \u2018Feelings\u2019 (8.49%); from the \u2018Perception of job performance\u2019 domain (12.58%), \u2018Workload\u2019 (4.40%); from the \u2018Perception of management\u2019 domain (9.12%), \u2018Management attitude\u2019 (4.72%); and, from the \u2018Communication in the surgical environment\u2019 domain (9.12%), \u2018Communication\u2019 (5.97%). What was said by the study participants is shown in Table 8.99%, \u2018CHaving presented the quantitative and qualitative results, the intersections between the approaches will now be shown, indicating the convergences and divergences Table . The firEvaluation of the findings from both approaches revealed divergences: from the quantitative data, the most positive domain for patient safety climate was \u2018Communication in the surgical environment\u2019 (77.91) while, from the qualitative data, this domain was the least prevalent in mentions (9.12%).However, convergence was also found: the themes that made up the \u2018Perception of job performance\u2019 domain were present in other categories of the qualitative analysis, such as \u2018COVID-19 Protocol\u2019 and \u2018Feelings\u2019. This was also the lowest-scoring domain in the quantitative analysis (23.60) and accordingly needs improvement and investment to benefit patient safety. It was mentioned in relation to various categories in the qualitative analysis, however, indicating that the study participants also recognised these needs.From the quantitative data, the domain with the most positive patient safety climate score was \u2018Communication in the surgical environment\u2019 and, from the qualitative data, the predominant domain was \u2018Working conditions\u2019. There was thus divergence between the findings from statistical analysis and those from the declarations of the multi-professional team in the study setting.The surgical environment has a particular, complex culture of its own, in which its personnel experience daily stressful situations that can culminate in conflicts in a tense work environment, where relations among different professions can be difficult. In view of this, it is imperative to adopt strategies for negotiation, communication and teamwork with a view to managing possibly conflictual situations .Surgical centre team meetings can be used as a management strategy for getting closer to the realities of the various members of the team, learning their difficulties and thus fostering action focused on strengthening related solutions . EffectiManagement involvement is fundamental to disseminating safety climate and consequently to evaluating, planning and implementing measures for improvement . A climaWhen combating an enemy like Covid-19, unity, collaboration between health teams, good communication and safety measures were extremely necessary. That given, the literature stresses that those measures should be permanent and not temporary, and thus contribute to patient safety .The \u201cSwiss cheese\u201d theory contributes to the feasibility of a safer care environment by encouraging increasingly effective barriers to prevent the metaphorical holes in the cheese from lining up. The idea of this is that, for a more mature safety climate for surgical patients, communication must improve between personnel at all levels of the hierarchy and punitive culture must be abandoned in favour of learning from error .The finding that the \u2018Working conditions\u2019 domain was the most prevalent in the qualitative data contributed to characterising the data collection period further: at that time, the health care staffs found themselves terrified by unknowns, learning to deal with situations day after day and, as frontline personnel in the endeavour to combat the Covid-19 pandemic, worried by issues of patient and personal safety , 31.Meanwhile, there was convergence with the \u2018Perception of job performance\u2019 domain; although this returned the lowest patient safety climate score (23.60), it was present in some of the qualitative categories. Remembering that this has to do with the impact of fatigue and overwork on job performance, it can be seen that these factors were present in the participants\u2019 declarations in the \u2018COVID-19 Protocol\u2019 and \u2018Feelings\u2019 categories created for analysis of the qualitative data.The challenges involved in installing Covid-19 control and prevention protocols caused participants to report fatigue during work; that is, health care personnel recognised fatigue and long working hours to be factors impairing job performance, resulting in loss of efficiency and productivity . A studyAs nursing is considered to form the backbone of health services, more investment must be applied to strategies to identify the psychosocial needs and situations of emotional vulnerability affecting these health care professionals . Fear isThe pandemic setting intensified workplace stress and had adverse effects on health care personnel\u2019s quality of life. This may burden the system through illness, absenteeism and leave for health treatment . MirandaIn view of these findings, it is important that management propose measures for continued professional development of health care teams and modify their perceptions of their importance to care and their fundamental role in patient safety, as well as stimulating a positive safety climate in the institution.As regards sociodemographic and job characteristics, the male sex predominated (50.69%) and, by profession, the largest team in this study was Medicine (63.89%). Standing against that finding, the nursing team accounted for 78.1% of the participants , as in SIn the job position category, surgical technicians and circulating nurses were found to predominate 26.39%). In this unit, these are nursing technicians and auxiliaries, as corroborated by other studies , 39. Nur%. In thiThe mean overall SAQ/OR score was 61.94, revealing that the staff perceived a neutral safety climate in the work environment. This suggests a considerable lack of initiative to improve these professionals\u2019 perceptions of the safety climate, such as by periodical educational interventions, which are considered of prime importance to improving perceptions of safety climate [It is important to note that patient safety issues have financial, social and psychological impacts by causing, for example, increased health costs and expenditures, emotional distress, pain and temporary or permanent interruption of work, which can affect both patient and institution . A fragiIn the pandemic context, health care personnel become preoccupied with workloads, the increasing complexity of the care to be provided and the limited resources available. Accordingly, Covid-19 is expected to have adverse impacts on quality of care and patient safety, making adjustments necessary in health institutions and posing the need to develop evidence-based strategies for reducing possible incidents .In the evaluation of the six domains that make up the SAQ/OR, the findings indicate that only the \u2018Communication in the surgical environment\u2019 domain (77.91) was perceived by both groups of professionals as being positive as regards safety attitudes in the work environment, which matches the finding of a study by . It is aCommunication is of the utmost importance in the surgical setting, so much so that one of the six essential goals for safe surgery is effective communication among the surgical team . In the The lowest-scoring domain (23.60) was \u2018Perception of job performance\u2019, which contributed least to the patient safety climate. Studies in southern Brazil  and at aA current study that examined the impact of Covid-19 on nursing work environments and patient safety culture, considering a period prior to the pandemic and then another after the third critical period of Covid-19, found that most patient safety dimensions were weak and needed to improve. That same study found that only the dimension concerned with teamwork in the units was regarded positively prior to the pandemic and after the third critical period of Covid-19 .Given that the positive responses on patient safety were significantly associated with the quality of the work environment, it is hoped that continuous investment in working conditions and promotion of an open and participatory safety culture will improve the quality of health personnel\u2019s work environment.In the qualitative analysis, from the \u2018working conditions\u2019 domain, which reflects the quality of the work environment, there emerged the category \u2018Covid-19 protocol\u2019. This made it possible to learn how the participants regarded the issues with the protocol established to address Covid-19.Health institutions faced with the pandemic needed to update with a view to improving knowledge of Covid-19 in order to provide appropriate, quality care . In thisIn the \u2018Safety climate\u2019 domain, where it is possible to learn staff perceptions of patient safety, the salient category was \u2018Patient safety\u2019. The interview participants perceived a commitment to patient safety on the part of the surgical team. All personnel, particularly those occupying positions at the head of the organisation, must make a priority of heightening patient safety and of implementing measures for that purpose, so that positive patient safety results are seen in the work process .In the surgical centre, there is the possibility of infection from patient to staff, but also from staff to patient, considering airway manipulation during anaesthesiology procedures and the difficulties in communication between teams . It is iIn the \u2018Perception of stress\u2019 domain, which considers how staff recognise the influence of stress factors on work routines, the predominant category was \u2018Feelings\u2019. Besides the risk of contamination, the Covid-19 pandemic demanded greater attention to the health of health care workers, who commonly display symptoms such as anxiety, depression, loss of sleep quality, fear of being infected or of infecting relatives and also burnout from overwork , 48.Worry about the possibility of contaminating family members is a highly significant psychosocial risk . During Knowing these conditions helps health institutions identify and develop measures to promote, treat and rehabilitate health personnel psychosocially . It is iThe \u2018Perception of professional performance\u2019 domain addresses the impact of workload and fatigue on professional performance. The category chosen to display the findings was \u2018Workload\u2019. Frontline workers combating Covid-19 had their working day modified by overtime and the pace of work . OvertimThere is a significant correlation between patient safety culture and work environment, burnout, depersonalisation and personal relations; when allied to fatigue, these influence patient safety . From thIt is indispensable for health care facilities not to make a priority of longer working days for health care personnel, particularly during pandemics . HoweverIn outbreaks and pandemics, it is common for health care workers to work longer hours, with no breaks and under heavy pressure, which leads them to fatigue and burnout . Such loThe \u2018Perception of management\u2019 domain considers staff approval for management attitudes in relation to the patient safety climate. The most prevalent category was \u2018Management attitude\u2019. From their declarations, the staff felt safeguarded by the health care facility\u2019s management. In workers\u2019 health care, it is important that personnel feel truly supported and not stigmatised . AttentiHowever, participants reported a lack of information from management, which caused insecurity and uncertainty. Communication is listed in the scientific literature as a strategy for health care personnel to cope with stress while combating Covid-19. This includes leaders\u2019 circulating success stories and constantly updating information on the local situations in the pandemic , 55.Accordingly, it is necessary to invest in quality information on health care workers\u2019 health, so as to measure health indicators appropriately and to guide workplace surveillance and inspection activities to support management decision making, favour health care workplace safety and contribute to health research .Lastly, the \u2018Communication in the surgical environment\u2019 domain addresses patient safety-related information shared among health personnel. In this domain, the category \u2018Communication\u2019 emerged. The interview participants stated the need for more comprehensive communication using, for example, meetings as a strategy. Surgical centre team meetings can be used as a management strategy for getting closer to the realities of the various members of the team, learning their difficulties and thus fostering action focused on strengthening related solutions . EffectiIt is imperative that staff enjoy relations of trust with management and their peers so that there can be positive communication in all directions. This recommends flexible arrangements that help establish effective channels of communication among different levels of the hierarchy, which can influence error reporting without degrading established relations .The study\u2019s practical contributions to surgical patient care quality and safety during the Covid-19 pandemic are directed to contributing thinking with a view to improvements focused on safe care with quality for surgical patients and, through the findings, to provide a basis for developing educational measures and interventions. Lastly, for science, particularly at a critical time worldwide, the study points to the need to foster research of this kind to encourage a mature, positive patient safety climate to develop and spread in healthcare environments.Despite the importance of its findings, the study has limitations, including particularly its being restricted to the surgical centre of one university hospital, the possibility that some interview script question may have induced response bias and the researcher\u2019s relationship with the study setting. Accordingly, it is suggested that further studies be conducted in other institutions in order to validate and generalise the findings.From the findings and the intersection between quantitative and qualitative data, the proposed study objective was achieved, in that it was possible to examine the surgical centre safety climate from the perceptions of the multi-professional staff.These diverged in that, in the quantitative approach, the highest-scoring domain was \u2018Communication in the surgical environment\u2019, whereas the participants\u2019 declarations in the qualitative approach indicated \u2018Working conditions\u2019. Nonetheless, the data also converged: in the quantitative analysis, the \u2018Perception of job performance\u2019 domain revealed the need for improvement measures and also permeated important categories and was often mentioned in the participants\u2019 declarations in the qualitative analysis, thus demonstrating that the health care personnel understood the effects of fatigue and overwork on job activities and the need for investment to modify that situation. That finding was achieved only after examining the intersections between the quantitative and qualitative data.It is suggested that further studies be conducted worldwide to explore the subject in depth, particularly in a variety of surgical settings and using mixed-method study designs, so as to permit future comparisons and strengthen the patient safety climate. Accordingly, it is hoped that this study can contribute with a view to improving patient safety in the surgical environment, so as to encourage evidence-based educational interventions to strengthen the safety climate and foster more assertive communication among health care personnel working in the surgical centre at all levels of the hierarchy."}
+{"text": "Newborn screening for congenital hypothyroidism (CH) has dramatically improved the neurocognitive outcomes for newborns with a confirmed positive screening test result. However, screening yields a small number of false positive and false negative results. This report describes the first known case of familial dysalbuminaemic hyperthyroxinaemia presenting with a positive newborn thyroid stimulating hormone screen. This condition is characterized by artefactually elevated free tetraiodothyronine (T4) and triiodothyronine (T3) levels due to increased albumin binding and subsequent dissociation during laboratory assays but normal true free thyroid hormone and thyroid stimulating hormone (TSH) levels in a clinically euthyroid subject. This highlights the need to take care when attributing clinical significance to discordant results. Newborn screening for congenital hypothyroidism has dramatically improved the neurocognitive outcomes for newborns with a confirmed positive screening test result. However, screening yields a small number of false positive and false negative results. This report describes the first known case of familial dysalbuminaemic hyperthyroxinaemia presenting with a positive newborn screen and highlights the need to take care when attributing clinical significance to discordant results.T4 and T3 are hydrophobic and require plasma transport proteins to get to target organs. The majority of T4 and T3 hormones are bound to three thyroid hormone (TH) transport proteins: thyroxin-binding globulin (TBG), transthyretin (TTR) and human serum albumin . Most TH binding is to TBG (75%), followed by TTR 15\u201320%), with albumin binding the least (5% of T4 and 20% of T3). Albumin is the most abundant protein in circulation and has more binding sites for TH than the others, but it has the lowest binding affinity [5\u201320%, wiFamilial dysalbuminaemic hyperthyroxinaemia (FDH) is caused by a missense mutation in the albumin (ALB) gene on chromosome 4, which causes a subtle change in the structure of albumin and increases its binding affinity for T4 or T3 [Baby A, a South Korean female, was referred for evaluation of possible congenital hypothyroidism at 4 days of age with elevated TSH on a newborn metabolic screening. Baby A was born at 37 + 5 weeks gestation via normal vaginal delivery. The pregnancy was unremarkable other than a maternal diagnosis of subclinical hypothyroidism with a serum TSH of 2.7 mU/L in the first trimester, for which the mother received 25 micrograms of thyroxine daily throughout the pregnancy . Thyroid antibody testing was not performed. Baby A\u2019s elder brother had a positive newborn screen for congenital hypothyroidism in South Korea but was never treated due to normal follow-up TFTs.Baby A\u2019s newborn screening TSH level was 22 mU/L in whole blood on a dried blood spot sample collected at 32 h of age. This met New Zealand criteria for a positive screen and direct referral for diagnostic assessment  . A bloodDue to the discordant TFTs, the following additional tests were performed to rule out interference in the biochemistry analyser and other artefacts: precipitation of antibodies and other large molecules/complexes with polyethylene glycol, treatment with heterophile antibody-blocking reagent, and serial dilution with normal human serum. In all cases, there was no gain or loss of recovery of TSH or free T4, suggesting that the unusual TFTs were not explained by the presence of macro-TSH, the presence of antibodies, or other factors that interfere with the analytical process. The free T4 analysed by Siemens Atellica was near the upper reference limit, while the total T4 on a Beckman Coulter immunoassay on day 15 was significantly higher than the upper reference limit .The initial differential diagnoses included thyroid hormone resistance and a thyroid hormone transport protein variant. Albumin analysis was then performed by liquid chromatography time-of-flight mass spectrometry (LC-TOF-MS) to investigate possible TH-binding variants. Apart from expected albumin  and cysteinylated albumin peaks , there were also clear peaks approximately 19 Da below the wild-type albumin peaks , suggesting heterozygosity for a normal and a variant albumin see .Baby A\u2019s genetic testing confirmed the diagnosis of FDH with heterozygosity of the NM_000477.5 (ALB):c.725G > A (p.Arg242His) variant in the albumin gene, or Arg218His in protein nomenclature, thus confirming the diagnosis of FDH.This report describes a well-recognised abnormality in albumin causing abnormal protein binding of T4 that has never been considered before in the interpretation of discordant follow-up test results of a newborn screen for CH. The elevated newborn screening dried blood spot TSH level suggests that this abnormality led to a transient, true reduction in free T4, which rapidly normalised postnatally. This is a previously unrecognised cause of a positive newborn TSH screening test. This case was puzzling, as while in-vivo free T4 levels are normal, as evidenced by a rapidly normalized TSH level, many commercial free T4 assays will produce artefactually elevated free T4 results in this condition.FDH is an autosomal dominant condition affecting the binding affinity of thyroid hormones to albumin. This results in elevated total T3 or T4 levels but normal free T3, free T4, and TSH levels . It is tThis condition can be difficult to detect because despite the free and biologically active TH levels being normal, the testing process causes dissociation of TH from albumin. This results in the measurement of both free TH and a fraction of the bound TH, resulting in artefactual elevation of free T4 . DissociMost clinical laboratories use indirect free TH quantitation methods, as they are faster, simpler, and less expensive than direct methods, such as equilibrium dialysis or ultracentrifugation. Indirect quantitation of free TH assumes the given fraction of measured free TH has come from the bound fraction and adjusts accordingly. This works well for the vast majority of patients. However, in FDH, a much larger quantity of free TH is bound to albumin and dissociates during analysis, producing spurious increased results. If a direct method were available in this case, normal free T4 results would be expected.Additionally, albumin can be analysed by liquid chromatography time-of-flight mass spectrometry (LC-TOF-MS) to clarify the cause of the discordant results. The masses of intact albumin can be assessed and compared to either a normal albumin or a patient with FDH. In the context of discordant TFTs, based on literature and local experience, a mass change of approximately \u221219 Da is highly suggestive of FDH due to Arg218His, whereas the presence of normal masses alone is consistent with normal albumin . AlthougThe limitations of this case report include the fact that only partial TFT results were available in family members. FDH is inherited in an autosomal dominant manner, and we suspect that the sibling\u2019s positive newborn screen may also have been caused by FDH. However, he was born overseas, and the details of his newborn screen and diagnostic TFTs were not accessible to us. The presence of maternal thyroid antibodies is an alternative explanation for a transient neonatal increase in TSH, although this invokes a second unproven pathology. Furthermore, reference intervals for TSH levels during pregnancy are unclear and assay dependent , and we This case highlights the need for caution in interpreting positive newborn screening results and discordant thyroid function tests, particularly when results don\u2019t fit a typical pattern and the associated clinical picture. FDH is most easily diagnosed by genetic testing for the albumin variant or alternatively can be assayed by liquid chromatography time-of-flight mass spectrometry (LC-TOF-MS) where available."}
+{"text": "This poses an interesting clinical question as papilledema is defined by elevated intracranial pressure. The ophthalmic defects of this patient with normal intracranial pressure and CM1 are explored in this report.Chiari malformation 1 (CM1) is defined as a herniation of encephalon matter through the base of the skull. The amount of herniation is cited as greater than 3 mm or 5 mm, depending on the source of literature. We report a rare case of a 55-year-old male initially presenting with bilateral papilledema and monocular right lower quadrantanopia, found to have CM1. An MRI confirmed\u00a04.87 mm herniation of the cerebellar tonsils at the foramen magnum, and he was diagnosed with CM1. He was later found to have a normal opening pressure on lumbar puncture at 10 cm H Chiari malformation (CM) is a condition caused by the movement of encephalon matter outside the skull and into the spinal cord, as described by Hans Chiari\u00a0in 1891 . Types IThe patient is a 55-year-old man with a past medical history of vitiligo, resolved childhood heart murmur, and hyperlipidemia. He noticed diminished peripheral vision in the right eye and mild right-sided headache over the parietal and apex of his head after a hike. The headache later resolved. Both an optometrist and an ophthalmologist diagnosed him with papilledema, with the right eye showing more pronounced symptoms. On presentation, he denied signs of increased intracranial pressure, including nausea, vomiting, and dizziness. He also reported no changes in hearing or coordination and had no history of trauma. Neurological evaluation revealed right monocular right lower quadrantanopia without other focal deficits. Color vision was intact, and his mental status was normal. Cranial nerves II-XII were normal, with no pronator drift, full strength throughout, normal light touch, normal finger-to-nose testing, and alternating hand movements. The patient did not have scoliosis.2O, with cerebrospinal fluid (CSF) analysis showing clear and colorless fluid with 5 WBC/mm3, 250 RBC/mm3, glucose levels of 68 mg/dl, and protein levels of 42 mg/dl. These findings indicate normal intracranial pressure. Thus, this patient was diagnosed with Chiari I malformation, presenting with a rare symptomology of unilateral peripheral visual field defects and bilateral optic disk swelling with normal intracranial pressure. The patient was advised to follow up in the outpatient neurology and ophthalmology clinic.Non-contrast computed tomography (CT) of the head was negative for hemorrhage, herniation, mass, or hydrocephalus. Magnetic resonance venogram (MRV) was negative for infarction. MRI\u00a0of the brain with and without contrast was positive for mild small vessel disease in the periventricular area, partially empty sella, and mild ectopia of bilateral cerebellar tonsils 4.87 mm below the foramen magnum with crowding Figure . These MThe presentation of CM1\u00a0ranges from asymptomatic to severe with tinnitus, vertigo, ataxia, headache, myelopathy, neck pain, nausea, and vomiting . It can This is a unique case of CM1 due to the atypical presentation of symptomatology of peripheral visual field defects with bilateral optic swelling in the absence of hydrocephalus or elevated intracranial pressure on the lumbar puncture. In addition, this is an uncommon age of presentation at 55 years old. Very few cases in literature document this sign in CM1. One case reported papilledema, bilateral anterior uveitis, and elevated ICP without hydrocephalus in a 16-year-old with CM1 . They weIt is unexpected to observe optic disk swelling in the presence of normal intracranial pressure, such as in this patient with an ICP of 10 mmHg. Papilledema, defined as swelling of the optic disk, typically occurs due to increased pressure on the optic nerve resulting in edema, which can be visualized through fundoscopy. Increased pressure on the optic nerve could be from elevated intracranial pressure, or infiltrative or compressive processes, such as tumors . In the A partial empty sella sign may be an incidental finding. However, we must, at the least, consider this as a pathology prior to dismissing it. The patient in this case report possesses an empty sella sign with ophthalmic defects and a normal pressure on lumbar puncture, which may make normal pressure pseudotumor cerebri a potential diagnosis. This finding was characterized in a case series, describing an empty sella sign on MRI, an average lumbar pressure of 11 mmHg, and ophthalmic defects such as horizontal diplopia, bilateral abducens palsy, optic nerve swelling on fundoscopy, and an enlarged blind spot ,11.The case involves an older patient presenting\u00a0with bilateral optic disk swelling and peripheral visual field defects, without the typical symptoms of increased intracranial pressure or hydrocephalus as the initial presentation of Chiari I malformation. Diagnostic imaging confirmed mild cerebellar ectopia consistent with CM1. This presentation is especially paradoxical due to the presence of bilateral optic disk swelling with normal opening pressure on lumbar puncture. Future efforts should be made to document patients with atypical presentations of CM to help understand the range of symptomology and the true incidence of this pathology."}
+{"text": "The widespread HIV epidemic in Ukraine is concentrated among people who inject drugs (PWID), making access to sterile injection paraphernalia (SIP) like sterile needles and syringes a critical method of HIV/AIDS prevention; however, the Russian invasion has threatened to disrupt the operations of syringe services programs (SSPs), creating a risk of HIV outbreaks among PWID.We conducted 10 semi-structured interviews with outreach workers from SSPs. Interviews were purposively sampled to cover three prototypic regions of Ukraine: temporarily Russian-controlled, frontline, and destination. Qualitative results from interviews were then compared against a standardized, nationwide harm reduction database.We found that the Russian invasion triggered both supply and demand challenges for SSPs. Demand increased for all regions due to client transitions from pharmacies that closed to SSPs, increases in illicit drug use, greater client openness to NGO support, and displacement of clients to destination regions. Supply decreased for all areas (except for remote destination regions) due to battle-related barriers like curfews, roadblocks, and Internet disruptions; diminished deliveries of SIP and funding; and staff displacement. Time series plots of the number of unique clients accessing harm reduction services showed that an initial decrease in service provision occurred at the start of the war but that most regions recovered within several months except for Russian-controlled regions, which continued to provide services to fewer clients relative to previous years.To ensure continued scale-up of SIP and other HIV prevention services, the SyrEx database should be leveraged to serve as a streamlined harm reduction locator that can inform workers and clients of open site locations and other pertinent information. Ukraine has the second-largest HIV epidemic in Europe, with ~360,000 people with HIV  and mostSyringe services programs (SSPs) substantially reduce risk of HIV transmission in PWID through distribution of sterile injection paraphernalia (SIP) along with disinfectant, provision of other tools for safe injection and linkage to opioid agonist therapies . Unlike Harm reduction organizations in Ukraine manage SSPs at both brick-and-mortar and through mobile outreach delivery . These eHere, we examine the extent to which SSPs in Ukraine evolved and altered their implementation strategies during the early phases of the Russian invasion. We used a previously described conflict framework that has been used to understand implementation of OAMT to analyze how different regions of the country responded to harm reduction during the early invasion , 15. SpeIn this mixed methods study, we employed qualitative analysis of semi-structured interviews and quantitative analysis of harm reduction data to evaluate the impact of the Russian invasion on SSPs.Russian-controlled, or areas where Russian forces had taken a form of control  over part of the region; frontline, or areas that were exposed to battle-related conflict but were not Russian-controlled; and destination, or areas that displaced persons had settled in for over 3\u2009months after coming from Russian-controlled and/or frontline regions. Though most destination regions were in western Ukraine , others were more central, like Poltava  with outreach workers from harm reduction organizations that manage SSPs within one of these three distinct regions. Interviews were completed in Russian by two coauthors, BMN and DJB, using video-conferencing software. Interview participants were purposively sampled based on the outlined theoretical framework to include similar numbers of participants from three conceptual areas based on proximity to the conflict: temporarily Russian-controlled, frontline, and destination regions. A semi-structured interview guide was used to direct questions, which probed participants on changes to supply and demand dynamics for SSPs following the Russian invasion of Ukraine  and where coding differed, a third coauthor arbitrated the disagreement. Quantitative harm reduction data were provided by the Alliance for Public Health from the SyrEx Database, which is used by harm reduction organizations in Ukraine to record the quantity of clients they serve and the services they provide. Data from SyrEx did not include the quantity of SIP distributed to clients. Therefore, we applied data detailing the number of unique clients among PWID who received at least one preventive service over time as a proxy for the quantity of clients that were specifically provided SIP. We generated time series of changes to this metric over time, from January 2019 through September 2022. Time series were analyzed for the nationwide aggregate of clients as well as for each Ukrainian oblast (province) in the study to visualize whether scale-up of SSP and harm reduction services was sustained over time, particularly after 24 February 2022.This study was approved by institutional reviews boards (IRBs) at Yale University and the Ukrainian Institute on Public Health Policy. Due to the low-risk nature of the study and the ongoing conflict in Ukraine, both IRBs endorsed collection of verbal consent from participants. All identifying information described by interview participants was redacted prior to transcription. All participants were financially compensated for interviews.N\u2009=\u20092), frontline (N\u2009=\u20095), and destination (N =\u20093) regions, as per our framework. From these interviews, we observed that SIP distribution during the war was disrupted due to changes in both supply and demand of these services, with considerable variation across regions.We interviewed 10 SSP workers from harm reduction organizations in Ukraine, located in temporarily Russian-controlled .Clients were displaced from Russian-controlled regions at the highest rate, though many remained, and newly displaced clients emerged in these regions as well. While controlled by Russia, the region was locked down, and individuals were not permitted to leave. Even prior to this closure, however, many clients chose to remain due to financial barriers and/or family issues.\u201cTo go somewhere else, you need to have money, you need to have some kind of profession to somehow live there, to have somewhere to go\u201d .In other cases, the release of prisoners due to an influx of pardons at the start of the Russian invasion caused an increase in the number of PWID in the region seeking access to HIV prevention services, including SSPs.\u201cThere were amnesties for people who were serving sentences in camps and prisons. And so, those who did not have time to leave settled [in Kherson]. It's hard for them. And so, they come to us\u201d .Prior to the invasion, many PWID preferred purchasing SIP in pharmacies rather than obtaining it from SSPs due to distrust of these programs. In the initial weeks of the invasion, however, many pharmacies across the country closed, leading PWID who had previously relied on pharmacies to pivot to SSPs, most of which continued operating.\u201cIn the past, if customers could get a job, they could get clean syringes or alcohol wipes and stuff like that [from pharmacies], but now our customers are out of work. Why? Because the small businesses, including pharmacies, have closed\u201d .Even as pharmacies reopened, PWID continued to rely on SSPs due to financial constraints. The costs of syringes in pharmacies increased substantially. In Russian-controlled and frontline regions, small business closures were common and often permanent, leading many PWID to lose their sole source of income. Even after pharmacies reopened following reductions in violence, many PWID lacked the funds to purchase SIP from pharmacies, resulting in increased reliance on SSPs. In response to the war, some pharmacies hiked up syringe prices, resulting in an additional financial barrier to access.\u201cIt was noticed that people who used to buy syringes themselves in pharmacies started saving money and coming to us to buy syringes. Because the money they were saving, they were using it to \u2026 buy drugs\u201d .\u201cAs far as I know, syringes have \u2026 doubled and tripled in price\u201d .Loss of employment also resulted in increased stress and desperation among SSP clients, many of whom had substance use disorders that made them more vulnerable to stress . This st\u201c[After the war] a lot of people have started to use bath salts. They are probably cheaper \u2026 We need to give out more [SIP than before]\u201d .\u201cDimedrol is now one of the most popular products in pharmacies\u2026 It seems like two-thirds of my clients now use Dimedrol, they say they need [larger] needles and syringes\u201d .\u201cI honestly know that a lot of new people use Dimedrol \u2026 They\u2019re now supplementing with it because they say that our methadone is weak\u201d .Aside from wartime stresses related to violence and financial issues, some clients increased their use of substances like synthetic cathinones and diphenhydramine during the war due to disruptions in OAMT services, such as when clinics closed and/or when OAMT clinics chose to reduce dosages of OAMT out of concern for insufficient medication supply.Limited access to methadone in clinics as well as to buprenorphine  likely led PWID to resort to purchase of illegal opioids like \u201cstreet methadone,\u201d another unknown opioid. Unlike methadone prescribed in clinics, which must be consumed orally, \u201cstreet methadone\u201d is usually injected by PWID, increasing the risk of transmission of HIV and HCV. This change in access has likely increased the demand for SIP among SSP clients.\u201c[OAMT clinics] started to give them [medication] less often. They used it all very quickly and then they sat without anything. Or they had to buy street drugs, and there was no money for that. Because if they could earn extra money somewhere else in peacetime, during the war it was very hard for them\u201d .\u201cIt was impossible to get to Kyiv from Boyarka [to receive OAMT]\u2026 there were problems for a month \u2026 [The clients] switched to \u2018street methadone\u2019 [because of these difficulties], and \u2018street methadone\u2019 is of very poor quality.\u201d .The severity of the Russian invasion, compounded by the desperation experienced by SSP clients, appeared to make clients more receptive to accept help from harm reduction organizations, despite client trust remaining limited.\u201cPeople are open now to any help. And they are more open to such services [like SSP], to any services that are provided\u201d .This change in client attitude further increased demand.Though SSP supply remained relatively constant in distal destination regions since the Russian invasion, it markedly decreased in both Russian-controlled frontline regions and proximal destination regions. In these areas, despite heightened demand for SIP, SSPs encountered major barriers to delivering those services to clients.As with clients, many SSP workers from harm reduction organizations were displaced due to conflict, with many moving to destination regions in Ukraine and/or neighboring countries in Europe. Though displaced staff generally continued to work remotely, SSPs lost in-person staffing capacity, which was critical for maintaining contact with clients and ensuring that clients had continuous and sufficient access to SIP. One participant reported that each SSP worker managed a specific set of clients within their organization; the loss of in-person staff in their organization led them to lose contact with several clients.\u201cEvery SSP worker has a client base. So, these are the people he\u2019s been in contact with for five to seven years. Because if we take our organization, we have people working for us for fourteen, fifteen, sixteen years \u2026 We suffer a lot when someone leaves our organization. These are people who have been working [with their clients] for a long time, who have [experience with them], and they know each other\u201d .Most organizations managed to hire new staff. For newly hired SSP workers, however, harm reduction organizations needed to ensure that they were adequately trained and had built sufficient trust with clients, a process that could take up to several months. The rapid turnover of staff led to substantial strains on harm reduction organizations, which reduced the supply capacity of SSPs by extension.\u201cFour people [left]. One joined us. Well, of course, the workload on the rest of the SSP workers increased. Work has increased for the entire staff, because if earlier four more people did it, now these four are gone, and the work remains\u201d .In some Russian-controlled and frontline regions, the imposition of roadblocks, curfews, and other battle-related disruptions inhibited organizations from routine distribution of SIP. The enforcement of strict curfews forced organizations to decrease their working hours, particularly during the most violent periods of the war.\u201cWhere I work, there was shelling nearby. There were troops landing nearby. And that\u2019s why we were handing out supplies very fast, two or three hours of work and that\u2019s it\u201d .For organizations operating in regions with restrictive roadblocks, they encountered major logistical barriers in their capacity to distribute SIP effectively. Prior to the invasion, most clients had accessed SIP at sites established by harm reduction organizations. With the installation of roadblocks, however, many of these sites became inaccessible to PWID who were afraid to interact with soldiers. Many clients in legal possession of methadone were arrested at these sites due to miscommunication and insufficient knowledge among soldiers. Furthermore, in Russian-controlled regions where Russia took control of the government, OAMT programs became illegal, causing additional challenges for PWID.Women are afraid to walk through roadblocks\u2026 A lot of guys are afraid too. Especially if they aren\u2019t feeling well, neuroses begin \u2026 they begin to behave suspiciously at the roadblock, and then they [the soldiers] undress them, look at injection marks, and check if there are any drugs. So, such people avoid roadblocks\u201d .\u201cThen moving around with methadone in your pocket, even if you had a permit, the territorial defense [staff] \u2026 didn't all know what methadone was. There were a lot of questions, there were a lot of arrests\u201d .\u201cClients are being seriously searched at roadblocks. Searches are done because they are suspicious persons\u2026 If a person [has been injecting drugs] for half of his lifetime, then he naturally looks suspicious\u201d .SSPs in regions with roadblocks attempted to accommodate clients by delivering SIP directly to clients\u2019 homes; however, SSP workers also encountered challenges at roadblocks while making these deliveries, further straining supply. Workers were subjected to inspections of their vehicles, precipitating confusion when soldiers found high quantities of SIP and harm reduction supplies. To avoid confiscation of supplies and/or arrest, SSP workers carried documentation to show they were authorized to make these deliveries. Still, some soldiers were reluctant to allow passage of supplies, delaying delivery. Participants reported, however, that once SSP workers had become acquainted with the soldiers working each roadblock, deliveries became smoother.\u201cPeople are afraid of [roadblocks]. But anyway, you get used to it. And the military itself no longer argues with us; in the beginning, it felt odd for them. \u2018How is it that he is passing through with such a large load of syringes?\u2019 Well, \u2018I'm a volunteer\u2019\u2014you show the sheet and explain the situation. We confirm everything and even go to the roadblock to sort it all out\u201d .In Russian-controlled areas, disruptions to cellular networks and the Internet further restricted the capacity of SSPs to efficiently supply services to clients. For many harm reduction organizations, maintaining contact with clients via messaging services was a crucial method for coordinating distribution of SIP. In Russian-controlled regions, this method of communication largely vanished when Russia took government control of the region. According to one SSP worker in Kherson, Russian occupiers disabled all Ukrainian networks and required individuals in the region to purchase cellular plans from Russian providers. Most clients lacked the funds to purchase these new cellular plans. For the limited number who could, they received brand-new phone numbers that harm reduction organizations did not have recorded in their databases, preventing contact with these individuals.In Russian-controlled and frontline regions, participants reported significant disruptions to delivery of funding and SIP, restricting SSPs\u2019 capacity to provide clients with sufficient supplies. Several SSPs cut the quantity of SIP that was distributed, reportedly due to issues with SIP delivery that arose due to conflict in these and neighboring areas.\u201cIn the region where I work, it turned out that we remained in Russian-controlled territory. We had blown up bridges around our town. And there was no access at all. No humanitarian aid vehicles could pass, I mean, absolutely nothing\u201d .Other participants reported that their organizations had substantially reduced the quantity of SIP distributed during the war due to delays in delivery of SIP to harm reduction organizations. This is confirmed in \u201cThe only issue now is quantity. We can\u2019t give more than five syringes to a person \u2026 Now it\u2019s two [per day], but before it was ten\u201d .\u201cIt is clear that [42 syringes per month] is not enough for [the clients]\u201d .duration of this trough in 2022 was longer than for prior years, with relatively low quantities of unique clients being recorded into April and May. This indicates likely evidence of war-related disruptions to these services.A quantitative analysis of the number of unique clients receiving at least one preventive service per month showed that, overall, the Russian invasion of Ukraine led to a reduction in the number of unique clients per month . BetweenBetween oblasts where we completed interviews, we found that for Russian-controlled regions , there was a decline in unique clients during the winter that was sustained through September 2022 . In fron\u201cMore work has been added \u2026 people from other regions came to Lviv. That led us to have a lot more work\u201d .This likely indicates that Lviv Oblast\u2019s reporting of clients into the SyrEx database was underestimated.According to most study participants, the SyRex database was used throughout the war and served as an important source for client linkage, particularly for displaced clients. All clients received de-identified numbers in the database along with a harm reduction card that granted them access to services across all of Ukraine.\u201cThose who came from one region to another, with the harm reduction card can receive services at any HIV prevention point. They can find out about prevention centers through the Internet \u2026 And they could [also] contact us directly\u201d .According to other participants, however, reporting to the SyRex database was not consistent, likely explaining the aberrant results observed in Lviv Oblast. For other participants, reporting to the SyRex database stalled amid the conflict due to the limited capacity of harm reduction organizations.There was no place to document; we didn\u2019t even have a place to print the documents out [like we did before the war] \u2026 We just did it all in a notebook because it was faster .When conflict receded in these regions, harm reduction organizations resumed reporting despite these delays.The Russian invasion of Ukraine poses a major, though not unprecedented, threat to Ukraine\u2019s capacity to continue its response to the HIV epidemic . As HIV Prior studies have identified similar disruptions to HIV treatment and prevention in Ukraine during the 2022 invasion, though none have focused specifically on harm reduction services to our knowledge. OAMT, a key HIV prevention strategy, has been significantly disrupted, with limited medication supply and rigid guidelines for treating patients that have exacerbated these disruptions , 15, 31.Our findings underline that it is highly important for harm reduction organizations to sufficiently insulate themselves from future disruptor events to ensure continued scale-up of SSPs. Disruption to SSP programs can be directly associated with increased transmission of HIV, making it crucial to continue service delivery . To dateOne successful strategy that enabled scale-up of OAMT for HIV prevention during the COVID-19 pandemic  and agaiSyrEx databases and other methods of data collaboration are therefore crucial for ensuring rapid scale-up during and after disruptor events. First, as proposed by Altice et al.  and our Non-governmental organizations (NGOs) that provide all of the harm reduction activities in the country have substantially contributed to cost-effective reductions in transmission of HIV and HCV . To sustThe lessons learned from Ukraine have important applications to other settings, especially where HIV and drug use epidemics are intertwined. First, the shift from opioids to stimulants and other non-opioids based on changes in drug supply, requires that SSPs enhance their services during conflict as OAMT has a limited role. Second, the types of settings that are at risk for such conflicts outside Ukraine are especially tenuous. For example, Russia has already annexed parts of Moldova and Georgia, both with inter-related HIV and opioid epidemics. Therefore, it is crucial for such settings to create preparedness plans for addressing future conflicts . Such coThough this study attempts to summarize the response of harm reduction organizations to barriers during the Russian invasion, it may not provide the full picture of the situation in Ukraine. Rather, it is based on the limited, available quantitative data and in-depth interviews that were performed with purposively selected participants across all regions of interest. Of note, this study does not include data from the Donetsk, Luhansk, or Crimea regions. This was due largely to logistical constraints, as it is very difficult to establish contact with SSPs and/or clients in these areas, which have been under Russian occupation since 2014. Moreover, this study was more focused on the specific response to the 2022 Russian invasion rather than the 2014 invasion.Maintenance and scale-up of SSPs in Ukraine is critical to preventing the spread of HIV and HCV for PWID. These programs not only distribute sterile SIP but also support and navigate clients to other services like OAMT that are crucial for overdose prevention and primary and secondary HIV prevention. Findings here demonstrate that the war has led to pressing and disparate needs across three different zones within Ukraine\u2014Russian-controlled, frontline, and destination. A targeted response should be tailored to each of these prototypical regions with an effective implementation strategy to ensure service continuity. Throughout the country, a national response that adheres to the supply and demand needs of SSPs for each harm reduction organization is critical to sustaining scale-up of SSPs despite the Russian invasion. Mobilization of existing resources like the SyrEx database can be particularly helpful for ensuring an improved response and recovery in the SSP space. These lessons from Ukraine should be applied across other countries that are similarly encountering concurrent HIV and drug injection epidemics.https://doi.org/10.17605/OSF.IO/BF4SK.The datasets presented in this study can be found in an online repository. The repository can be accessed at the following link: The studies involving humans were approved by Yale University and the Ukrainian Institute of Public Health Policy. The studies were conducted in accordance with the local legislation and institutional requirements. The ethics committee/institutional review board waived the requirement of written informed consent for participation from the participants or the participants\u2019 legal guardians/next of kin because due to the low-risk nature of the study and the ongoing conflict in Ukraine, both IRBs endorsed collection of verbal consent from participants.BN: writing-original draft, conceptualization, methodology, investigation, visualization, and formal analysis DB: conceptualization, methodology, investigation, visualization, writing-review and editing, and formal analysis IP: conceptualization, validation, and resources RI: validation and methodology ZI: validation, resources, and conceptualization. FA: supervision, conceptualization, and validation. All authors contributed to the article and approved the submitted version."
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