In addition, this study presents a multivariate correlation analysis of experimental parameters. The treatment using RF plasma O2 aimed to increase the hydrophilic character of the raw fabric and adherence of paste-based polymeric on polyvinyl alcohol (PVA) matrix and nickel (Ni), silver (Ag) or copper (Cu) microparticles. The purpose of the research was to develop electroconductive textiles for flexible electrodes, smart materials using a clean technology such as radiofrequency (RF) plasma O2 to obtain a hydrophilic surface with zero wastewater and reduced chemicals and carbon footprint. To achieve the foreseen results, we used advanced functionalization technologies such as RF plasma O2, followed by scraping a thin film of conductive paste-based Ni, Ag or Cu microparticles, and multivariate correlation methods to observe the dependence between parameters involved (dependent and independent variables). Overall, the fabrics treated in plasma with O2 using a kHz or MHz generator and power 100–200 W present an excellent hydrophilic character obtained in 3 min. After RF O2 plasma functionalization, a thin film based on polymeric matrix PVA and Ni microparticles have been deposited on the fabric surface to obtain electroconductive materials.This paper presents a study concerning the preliminary treatments in radiofrequency (RF)oxygen (O Samples A1–A6 have been developed based on 3 weaved structures from 100% cotton yarns: twill 2/2 for samples A1 and A4, twill 3/1 for samples A2 and A5, and plain weave 1/1 with cotton yarns Nm 20/2 on weft for samples A3 and A6.Several raw fabrics presented in 2 to avoid classical cleaning treatments that harm air and water through intensive pollution and high chemical consumption. From 3 Ω and are not dependent on the surface type (hydrophilic or hydrophobic). In the case of the samples A1 and A4 having initial surfaces that are hydrophilic and hydrophobic, respectively, and treated with a conductive paste-based Cu, we can observe that the surface resistances after crosslinking are 1012 Ω and 1010 Ω, respectively, which means that sample A1 has an insulator character, and sample A4 has an antistatic character.The purpose of the experiments was to compare the classical alkaline boiling-bleaching followed by conductive paste scraping (samples A1–A3), the classical alkaline boiling-bleaching followed by hydrophobization using NUVA TTC and conductive paste scraping (samples A4–A6) and the conductive paste deposition after functionalization in RF plasma OThe surface morphology was obtained by scan electron microscopy Quanta 200 SEM  using 2000× magnification.2 and samples coated with conductive paste-based polymeric matrix and Ni are presented in 2 and RF1 MHz generator and power 200 W in comparison with the sample treated using RF2 kHz generator and power 50 W  microparticles. In -Sample 18 from -Sample 2 from -Sample 10 from The chemical composition was investigated using the ESD (Energy Dispersive Spectroscopy) analysis and elemental mapping overlay for samples 18, 2 and 10  performe1 generator and power 200 W (2 followed by coating with Ni-based paste (2 generator and power 100 W (2 followed by coating with Ni-based paste (2 using RF1 generator and 200 W power) than 2 using RF2 generator and 100 W power).er 200 W  and treaed paste . Figure ed paste  and the ed paste  for samper 100 W  and treaed paste . The proed paste  are becaed paste  on the E2 and 50 W power in comparison with the samples treated in RF plasma using RF1 generator and power 200 W , in frequency (using two different RF generators having the frequencies 13.56 MHz (RF1) and 40 kHz (RF1)).The experiments in RF plasma O2 using the generator RF1 and power 200 W become hydrophilic. In addition, the samples treated in RF plasma O2 using the generator RF2 and power 100 W become hydrophilic.The samples treated in RF plasma O2 using the generator RF2 and power 50 W continued to be hydrophobic.On the other hand, the samples treated in RF plasma OESD and M, Pa, Pv and δ. The correlation coefficients were calculated for all samples preliminarily treated using RF plasma O2 with RF1 generator and power 200 W, RF2 and power 100 W, and RF2 and power 50 W:x, y represents the individual values of the variables x and y;x, y;xs, ys represents the standard deviation of all values x and y.The correlation between surface resistance after free drying or crosslinking (dependent variable) and other independent variables  was investigated by analyzing the correlation coefficient Pearson (1) between R1 and power 200 W, ESD free drying), after free drying at room temperature (18–20 °C) for 20–24 h, depending on the independent variables , the correlation coefficients between the RESD free drying and mass (2), air permeability (3), vapor permeability (4) and thickness (5) were calculated.In the case of the fabrics treated in RF plasma using RFhickness , air perhickness , vapor phickness , air perhickness , mass anhickness  and masshickness  to obserESD) after free drying and M, Pa, Pv and δ (ESD) and air permeability (Pa) and vapor permeability (Pv), it is a direct correlation, and this indicates that a textile fabric having higher values for Pa and Pv surface will generate the increase of surface resistance value. Moreover, between RESD and thickness and mass, it is an insignificant dependence.Analyzing the values of the correlation coefficients between surface resistance  and the black dots represent the appropriate points for RESD free drying depending on mass (M) and thickness (δ).ESD free drying and  and the black dots represent the fitting points for RESD free drying depending on air permeability (Pa) and thickness (δ).ESD free drying and  and the black dots represent the fitting points for RESD free drying depending on vapor permeability (Pv) and thickness (δ).ESD free drying and  and the black dots represent the fitting points for RESD free drying depending on air permeability (Pa) and vapor permeability (Pv).ESD free drying and  and the black dots represent the fitting points for RESD free drying depending on mass (M) and vapor permeability (Pv).ESD free drying and  and the black dots represent the fitting points for RESD free drying depending on mass (M) and air permeability (Pa).1 and power 200 W, ESD after crosslinking), after crosslinking, depending on the independent variables , the correlation coefficients between the RESD crosslinking and mass (6), air permeability (7), vapor permeability (8), and thickness (9) were calculated.In the case of the fabrics treated in RF plasma using RFhickness , air perhickness , vapor phickness , air perhickness , mass anhickness , mass anhickness ) to obseESD) after crosslinking and M, Pa, Pv and δ (ESD) and vapor permeability (Pv), thickness (δ) and mass (M), the correlation coefficients are negative, and the inverse correlation indicates that in the case of a textile fabric coated with a conductive paste-based Ni, the increase of the values for δ, Pv and M will generate a reduction of the RESD value. However, between RESD and Pa, the correlation coefficient is moderately positive, which means a direct insignificant dependence between these variables.By analysis of the values for correlation coefficients between surface resistance  and the black dots represent the fitting points for RESD after crosslinking depending on mass (M) and thickness (δ).ESD after crosslinking and  and the black dots represent the fitting points for RESD after crosslinking depending on air permeability (Pa) and thickness (δ).ESD after crosslinking and  and the black dots represent the fitting points for RESD after crosslinking depending on vapor permeability (Pa) and thickness (δ).ESD after crosslinking and  and the black dots represent the fitting points for RESD after crosslinking depending on air permeability (Pa) and vapor permeability (Pv).ESD after crosslinking and  and the black dots represent the fitting points for RESD after crosslinking depending on mass (M) and vapor permeability (Pv).ESD after crosslinking and  and the black dots represent the fitting points for RESD after crosslinking depending on mass (M) and air permeability (Pa).2 and power 100 W, ESD after crosslinking), after crosslinking, depending on the independent variables .In the case of the fabrics treated in RF plasma using RFhickness , air perhickness , vapor phickness , air perhickness , mass anhickness  and masshickness  to obser2 with RF2 generator and power 100 W, the correlation coefficients (10)–(13) have been calculated:In the case of the samples preliminarily treated using RF plasma OESD) and air permeability (Pa), there is a direct correlation, and these variables are in a direct proportionality relationship, and this indicates that the increase of the Pa value generates the increase of the RESD and reduction of the electrical conductivity. In addition, we can observe that between RESD and M, Pv and δ, the correlation coefficients are negative. The inverse correlation indicates that increasing the value of the parameters M, Pv and δ will reduce surface resistance and increase the conductivity.Analyzing the values of the correlation coefficient ESD after crosslinking and  and the black dots represent the fitting points for RESD after crosslinking depending on mass (M) and thickness (δ).ESD after crosslinking and  and the black dots represent the fitting points for RESD after crosslinking depending on air permeability (Pa) and thickness (δ).ESD after crosslinking and  and the black dots represent the fitting points for RESD after crosslinking depending on vapor permeability (Pv) and thickness (δ).ESD after crosslinking and  and the black dots represent the fitting points for RESD after crosslinking depending on air permeability (Pa) and vapor permeability (Pv).ESD after crosslinking and  and the black dots represent the fitting points for RESD after crosslinking depending on mass (M) and air permeability (Pa).ESD after crosslinking and  and the black dots represent the fitting points for RESD after crosslinking depending on mass (M) and vapor permeability (Pv).In 2 and power 50 W, ESD after crosslinking), after crosslinking, depending on the independent variables .In the case of the fabrics treated in RF plasma using RFhickness , air perhickness , vapor phickness , air perhickness , mass anhickness  and masshickness  to obser2 with RF2 generator and power 50 W, the correlation coefficients have been calculated (14)–(17):In the case of the samples preliminarily treated using RF plasma OESD after crosslinking and M, Pa, Pv and δ  and the black dots represent the fitting points for RESD after crosslinking depending on mass (M) and thickness (δ).ESD after crosslinking and  and the black dots represent the fitting points for RESD after crosslinking depending on air permeability (Pa) and thickness (δ).ESD after crosslinking and  and the black dots represent the fitting points for RESD after crosslinking depending on vapor permeability (Pv) and thickness (δ).ESD after crosslinking and  and the black dots represent the fitting points for RESD after crosslinking depending on air permeability (Pa) and vapor permeability (Pv).ESD after crosslinking and  and the black dots represent the fitting points for RESD after crosslinking depending on mass (M) and air permeability (Pa).ESD after crosslinking and  and the black dots represent the fitting points for RESD after crosslinking depending on mass (M) and vapor permeability (Pv).2 using RF1 generator at 13.56 MHz frequency and power 200 W and using RF2 generator at 40 kHz frequency and 100 W power present an excellent hydrophilic character only in 3 min compared to samples treated in RF plasma O2 using the RF2 generator at 40 kHz frequency and power 50 W.In conclusion, the samples treated in RF plasma OMoreover, in the case of samples functionalized in plasma and coated with PVA paste-based Ni particles, it was observed that the samples become conductive after drying at room temperature (18–20 °C) for 20–24 h.9 Ω to 1010 Ω, and the resurface resistance value is a specific value for antistatic fabrics.In addition, in the samples hydrophobized with NUVA TTC and treated with polymeric paste-based PVA matrix and Cu microparticles, after crosslinking, the surface resistance was increased ten times, from 10ESD after crosslinking and M, δ and Pv are negative in the case of the samples pretreated in RF plasma O2 using RF1 generator and 200 W power, and this indicates a strong inverse correlation and an inverse proportionality relationship between these variables. However, the correlation coefficient between RESD after crosslinking and Pa is positive, and this shows a direct correlation and direct proportionality between these variables and suggests that increasing the Pa  will generate the increasing of the RESD after crosslinking.Analyzing the correlation coefficients, we can conclude that the correlation coefficients between R2 using RF1 generator and power 200 W and RF2 generator and 100 W power, followed by coating with a thin film based on PVA matrix, organic solvents and Ni microparticles present a good conductivity, reduced surface resistance and can be used for sensors development.The fabrics that are treated in RF plasma OThe development of conductive textiles for flexible electrodes using clean technology such as RF plasma oxygen leads to decreasing the wastewater, chemicals, carbon footprint and pretreatment time.


Integration has become a major concern for governments, healthcare and aged care systems in many countries. However, the research on and implementation of integrated care in China started relatively late, and there is no review on the needs of older adults with regard to integrated care and the influencing factors. Therefore, this paper aims to provide a scoping review by searching, evaluating, and summarizing the Chinese and international literature on the need for and the factors influencing integrated care for older people. In addition, this review highlights evidence of the gap between China and the world in integrated care.Using a framework proposed by Arksey and O’Malley, a systematic search of 12 domestic and international databases was conducted. Of the 890 original studies retrieved, those that met the established inclusion criteria were screened and scored using the Ekman quality assessment tool. The qualitative description method was used to summarize the demand for integrated care for older adults and the influencing factors.A total of 49 papers were included. These studies were from eleven countries on five continents (most commonly China and the US) and were mostly cross-sectional quantitative studies that surveyed the integrated care needs of older people living in homes/communities or long-term care facilities. The analysis shows that existing research on the integrated care needs of older people in China adopts a single perspective and is inadequate and unsystematic in its assessment; the integrated care needs of older adults and the factors influencing them are multifaceted; and both in China and internationally, the community-home care scenario most consistently meets the needs and expectations of older adults.Although there is no uniform definition of integrated care in China or abroad and each country has its own national definition and system of integrated care, there are certain commonalities regarding the needs of older adults and the factors that influence them across countries. Our research reveals a gap between China and the international community in terms of integrated care. In recent years, the ageing trend has become prominent in China. Compared with European developed countries and the United States, China shows a unique ageing pattern of “Ageing before Affluence”, i.e., the rate of population ageing exceeds the rate of per capita income increase . AdditioYiyang Jiehe’—which means “health and social care combination” or “medical and older adults care combination”—has become synonymous with integrated care for older people [5In China, ‘r people 5. The Ch people 5 and desi people 5.Due to their rapid economic development and the early emergence of ageing problems, some developed countries established an institutionalized, large-scale, and increasingly mature integrated care model for older adults as early as the last century through pilots, practice and extension 8. The moYiyang Jiehe’, which is focused on exploring models of integrated care and their pathways to realization and involves four levels: nursing home, home, community and hospital [2medical-nursing combination’ are people-centred [To achieve the Global Strategy and Plan of Action for Active and Healthy Ageing, the World Health Organization (WHO) published the Integrated Caring for Older People (ICOPE) guidelines in 2017, which aim to provide community, primary and secondary health care providers with evidence-based recommendations for preventing, delaying or reversing physical and mental decline in older people . The WHOospital 2. The WHO-centred . Additio-centred 22.As the WHO-ICOPE has noted, the needs of older adults should be fully understood before providing integrated care services. However, as older people age, their health problems tend to become more chronic and complex, with multimorbidity, the coexistence of multiple chronic conditions, becoming the norm rather than the exception . Older a182627In recent years, research on integrated care in China has focused on the connotations of the model, the existing problems and the realization path, while less attention has been given to the cognition and demand of the older population with respect to integrated care 29. Older30Because the scoping review method is flexible and allows qualitative and quantitative research to be included, a variety of topics can be explored without restriction, such as the needs of patients and caregivers and the determinants of health. It can also be used to identify existing gaps in the literature. These gaps 33 are inThis study adopts the framework proposed by Arksey and O’Malley  and refiThe purpose of this review was noted above. At present, there is no uniform conclusion regarding ‘integrated care’, either domestically or internationally. For the purposes of this review, we have combined the WHO descriptions  with Leu“a process to overcome fragmentation, based on a person-centred approach that connects the health care system  with other human service systems  to improve outcomes  through different levels and sites of care and service delivery to meet people’s needs”.The review attempts to answer three research questions:In integrated care, what are the specific service needs of older adults?What factors  affect older adults’ needs of for integrated care services?In research and services on integrated care, what are the differences in the needs of older people in China and abroad and their influencing factors? What gaps exist?Published and unpublished (‘grey’) literature were identified based on a comprehensive three-step search strategy recommended by the Joanna Briggs Institute (JBI) systematic reviews . In the Table 1 for example search strategies from one database).Chinese search strategy: Five Chinese databases were used, namely, the China National Knowledge Infrastructure (CNKI), Wang Fang Data, VIP Chinese Database (VIP), Chinese Scientific Document Service System (CSCD) and Chinese Social Science Citation Index (CSSCI), were searched. Foreign language search strategy: Seven English databases were used, namely, Web of Science, Elsevier/Science Direct, MEDLINE, PubMed, Springer Link, EBSCO ASP, and The Cochrane Library. A manual search was also performed with Baidu Academic and Google search. We did not limit publication dates or language in the searches. We consulted a scientific librarian for the synonyms and search strategies for each database .Studies were selected through a two-step process using the selection criteria below. Two members of the research team independently reviewed the titles and abstracts of each study according to the literature inclusion criteria to complete the preliminary screening. The two team members then carefully read the full text of the articles and used the exclusion criteria to evaluate and complete the secondary screening. If there were differences of opinion, another researcher on the team was consulted to obtain a consistent final result. In addition, during this review, guided by the PRIMSA statement for systematic review protocols , a flowcThe inclusion criteria were as follows: (1) the needs of older adults for integrated care and the influencing factors were studied, and the relevant research results were summarized; (2) the study involved older adults aged ≥60 years; (3) the service location included in the literature could be any setting, including a medical or health institution, an older adult care institution, a community, or a home; and (4) any type of primary study  was performed.The exclusion criteria were (1) reviews, press releases, commentaries, conference briefs, and science fiction; (2) publications concerning older people’s perceived attitudes (acceptance or lack thereof rather than need) or satisfaction with integrated care; (3) duplicates or literature for which full-text information was unavailable; (4) literature that was less relevant to the content of the study; (5) literature that lacked either of the two themes of demand for or factors influencing integrated care; and (6) literature that was rated as low quality by the Ekman quality assessment tool .Although the scoping review did not have strict requirements for quality control of the literature, this study used the Ekman quality assessment tool to evaluate the quality of the included literature. The tool was initially used to evaluate studies on community health services insurance in low- and middle-income countries  and coveMicrosoft Excel software was used to extract data, and bibliometric methods were applied. The final information included the following three categories: (1) literature information: country, first author, publication year, journal name, journal level, article type, and research method; (2) key content: service venue, integrated care needs and influencing factors; and (3) survey information: survey population and sample size, recruitment and data collection methods, inclusion and exclusion criteria, survey duration, and related information. Charting should be an iterative process in which researchers continually extract data and update the data-charting form .Based on the description of Arksey and O’Malley  and LevaIn this scoping review, we sought information from older adults and their caregivers about integrated care needs and the factors influencing them in older adults. In terms of integrated care needs, we referred to the service content and framework in the “Guidelines for the Management of Integrated Medical Care and Care Institutions ”  issued bConsultation and member checking are optional when conducting scoping reviews . HoweverBy applying the search strategy, out of 898 citations retrieved, 110 were duplicates, leaving 788 for title/abstract and full text screening, resulting in 49 articles (24 articles in Chinese and 25 articles in English) included in this review. Of these, 25 are high quality (3 stars) [22540414243444546474849505152535455565758596061563646566676869707172737475767778798081828384The earliest journal article was published by Li  in 2004,2546474955586061626566676971727374757677787981828384404142444552535443574825474955626751536264717375254041424548505557637444594352535625464749555860616265666769717273767778798182838452We also found that the sample size of the included studies ranged from a minimum of 56 people  to a max4241435968592540414243444546485052535455566263646668707374757778824751575859607276808125496165676979After classifying and summarizing the actual integrated care needs of older people in the included literature, 8 categories of needs that make up the integrated care needs of older people emerged: basic life, medical and nursing, rehabilitation, ancillary, psycho-spiritual support, social participation, health education, and welfare and aid.Table 2 for a description of each subcategory).For basic life needs, there are 13 subcategories under this category, mapping 43 articles that cover nearly all the basic needs of older people in their daily lives. Of these needs, self-care, housekeeping and meal preparation were the most commonly identified service needs, covering 31, 25 and 19 articles in that order. Six articles referred to the sensory needs of older people in terms of sight, hearing and communication, while sensory needs and self-mobility were discussed simultaneously in 4 articles. However, both sleep aids and memory were only mentioned in 2 articles. The need for access to medical and nursing needs was a category that emerged most often in the studies reviewed; 17 subcategories and 45 articles were included. Of these, 21 articles explored the home health care needs of older people. Disease diagnosis and treatment (n = 18), regular physical examination (n = 14), skilled nursing (n = 11) and medication (n = 11) are all high-frequency needs. However, both emergency care and skin care were discussed in only 1 article each. In total, 14 articles explored the rehabilitation needs of older people, with 10 additional articles addressing rehabilitation and health care rather than rehabilitation guidance. Within the category of ancillary needs, six sub-categories and 19 articles were found, and a relatively high number of articles explored the four subcategories of transportation (n = 8), shopping (n = 5), older adults care hotline (n = 5), and managing money (n = 5); Among psycho-spiritual support needs, there were a large number of articles on spiritual solace, psychological counselling, and company, representing 12, 15, and 12 articles, respectively. The social participation needs category includes 10 subcategories, with cultural and entertainment service (CES) being the most discussed subcategory with 17 articles included. Among health education needs, both health guidance and health lectures appeared in 5 articles, while the three needs of health guidance, health lectures and health training were mentioned simultaneously in Liu’s  dissertaTable 3).After collating and summarizing the influencing factors obtained through various analytical methods in the included literature, it was found that the factors affecting the integrated care needs of older adults can be divided into six categories: demographic, personal, psychological, family, community, and social. Of these, 33 articles explored demographic factors, and the age (n = 19), gender (n = 11), educational attainment (n = 15), preretirement occupation (n = 7) and marital status (n = 9) of older adults were the main demographic factors that influenced integrated care needs. Among the personal factors, the ones with the highest number of articles were health and economic factors, including 37 articles and 23 articles, respectively. Psychological factors mainly included depression and mental state, and 6 articles were included. Both the family factors and the social factors constitute three subfactors each, with different elements under these subfactors. Family factors were highlighted, as the number of adult children (n = 9) and the living arrangements of older people (n = 9) were discussed more frequently. Studies outside of China identified more caregiver factors (including 3 articles), but only one Chinese study mentioned caregivers . In addiOur scoping review included 49 studies covering the integrated care needs of older people aged 60 and older in 11 countries. We noted a considerable degree of similarity in older adults’ needs and factors influencing across the countries, although there were a few clear differences. The integrated care needs of the older adults in each country comprise eight categories: basic life, medical and nursing, rehabilitation, ancillary, psycho-spiritual support, social participation, health education, and welfare and aid. The factors affecting these needs fall into six categories: demographic, personal, psychological, family, community and social. This review adds an important new perspective to the broader body of knowledge on coordinated person-centred integrated care research . In addiOur review found that many studies outside of China were more multifaceted and holistic in their investigations than those in China. First, in terms of research subjects, several international studies have studied the perspective of older people and their caregivers 43596880,435968556273744857596340535759We also found that foreign studies began to differentiate between met and unmet needs 2552575980 525759At present, empirical research on the need for and willingness to accept integrated care in China is neither up-to-date nor sufficient . The exiIn addition, as China’s integrated care needs assessment of the older population is not yet systematic, resulting in crossover and overlap among medical, nursing, health care and rehabilitation services, many scholars are accustomed to randomly combining these four types of services to conduct research, and there is no unified definition of integrated services . In ChinIn general, basic life care and medical-nursing care are the main focuses of integrated care for older adults. In the category of basic life needs, self-care, housekeeping and meal preparation were the services that emerged most often in the studies included in the review. The main reason for this result is that the deterioration of older adults’ physical functions and decrease in energy make it increasingly difficult for older adults to perform household chores . When fa41424350485940505255497678254042444552542552596343With improved material and economic conditions, the number of people requiring spiritual comfort and psychological counselling has increased . The old5162The factors influencing the need for integrated care in older people are broad and complex, involving several different aspects. Demographic characteristics and personal factors are the main factors influencing the integrated care needs of older people, with family and social factors ranking second, suggesting to some extent that the proportion of older people relying on their children to provide family care has declined. Due to globalization and the pursuit of self-fulfilment, geographical distance and life and work pressures make it difficult for young people to provide financial, emotional and caregiving support for their parents; hence, older Chinese people have changed their caregiving expectations, and the value of traditional filial piety that required absolute obedience from children has changed . Age, ge2Among family factors, older people with fewer children or who live alone show a greater need for integrated care 62, and t5654447Regardless of country or region, as the community and family are the most basic spheres of human life, people prefer to age at home or in the community rather than in an institution 4591. Our45Moreover, our literature search also revealed that there were few studies conducted through “LTC institutions” as care scenarios in China 4772768184,727681A strength of this study is the rigorous and systematic process of the literature search, which was conducted following Arksey and O’Malley’s guidelines . To our The review fulfilled the original objectives, answered the questions posed, and identified and summarized the specific needs of older adults in China and abroad for integrated care and the factors influencing them. There are both similarities and differences in the integrated care needs of older people at home and abroad. We also found a large gap between China and the international community in terms of integrated care and even the overall demand for older adult care services. The current research methods and perspectives on the demand for integrated care among older adults in China are still relatively homogeneous, inadequate and unstructured. The results of this review will not only serve as a reference for domestic and international researchers and policy makers but also clarify the direction of efforts and improvements. In brief, there is a long way to go in providing suitable integrated care for older adults in China.The additional file for this article can be found as follows:10.5334/ijic.5946.s1Appendix 1.Characteristics of the included studies.


Following the publication of this article , concern○Fig 1D, RKIP panel, between lanes 2 and 3.○Fig 2A DU145 RKIP panel, between lanes 1 and 2, as well as between lanes 2 and 3.○Fig 2A PC3 RKIP panel, between lanes 1 and 2, as well as between lanes 2 and 3.Vertical irregularities suggestive of splice lines were detected in the following panels:In Fig 2D, the PC3 PARP panel appears similar to the first two lanes of the right (DU145) panel, despite representing results obtained from different cell lines.○The first two lanes of the middle pY705 STAT3 panel appear similar to the first two lanes of the right pY705 STAT 3 panel. However, the corresponding Actin panel results for these lanes do not appear to match.○Lanes 3 and 4 in the middle panel appear similar to lanes 7 and 8 in the middle panel respectively, despite being used to represent different experimental conditions.○The last lane of the middle pY705 STAT3 panel appears similar to the last lane of the right pY705 STAT3 panel, despite being used to represent different experimental conditions.In Fig 4A, the following similarities were detected between lanes presented in the middle panel and lanes presented in the right panel:The authors have not provided the original images underlying the panels of concern.PLOS ONE Editors retract this article.In light of the concerns affecting multiple figure panels that question the integrity of the data, the ELM agreed with the retraction and apologises for the issues with the published article. BB did not agree with the retraction. SY, MD, SC-K, KB, TL, KCY, FA-M, EC, and DC either did not respond directly or could not be reached.


The large number of pathologies that position CCR5 as a central molecular determinant substantiates the studies aimed at understanding receptor-ligand interactions, as well as the development of compounds that efficiently block this receptor. This perspective focuses on CCR5 antagonism as the preferred landscape for therapeutic intervention, thus the receptor active site occupancy by known antagonists of different origins is overviewed. CCL5 is a natural agonist ligand for CCR5 and an extensively studied scaffold for CCR5 antagonists production through chemokine N-terminus modification. A retrospective 3D modeling analysis on recently developed CCL5 mutants and their contribution to enhanced anti-HIV-1 activity is reported here. These results allow us to prospect the development of conceptually novel amino acid substitutions outside the CCL5 N-terminus hotspot. CCR5 interaction improvement in regions distal to the chemokine N-terminus, as well as the stabilization of the chemokine hydrophobic core are strategies that influence binding affinity and stability beyond the agonist/antagonist dualism. Furthermore, the development of allosteric antagonists topologically remote from the orthosteric site  is an intriguing new avenue in GPCR druggability and thus a conceivable novel direction for CCR5 blockade. Ultimately, the three-dimensional structure elucidation of the interaction between various ligands and CCR5 helps illuminate the active site occupancy and mechanism of action. Staphylococcus aureus infections, cancer and atherosclerosis). In most of these pathologies, CCR5 blockade is a promising therapeutic avenue (S. aureus LukED toxin). Interestingly, both gp120 and LukED pathogenic engagement of CCR5 has been reported not to activate the receptor  (PDB ID: 5UIW)  was usedNext, we modeled on 5UIW the first 15 AA of CCR5 N-terminus from the NMR solution structure of monomeric CCL5 bound to a synthetic doubly sulfated CCR5 N-terminus peptide (PDB ID: 6FGP) . We usedA retrospective structure-guided analysis was conducted to examine the quality and effect the six AA differences between 5P7 CCL5 and CCL 5P12 5M exerted on the full CCR5 model. Structures were visualized and analyzed using the PyMOL software.CCR5 stabilization in an inactive conformation by the use of compounds capable to occupy the active site as antagonists may play a significant therapeutic role in several pathologies Figure 1Among the CCL5 derivatives efficiently blocking CCR5, the most potent HIV-1 entry inhibitors reported to date are CCL5 5P12 5M and CCL5 6P4 5M, a CCR5 antagonist and a superagonist, respectively . Five muThe CCL5 5M derivatives have been generated in the absence of the 3D structural details of the CCL5:CCR5 interaction interface, and were the result of previous findings derived from CCL5 short peptides with anti-HIV-1 activity . The cryThe CCR5 antagonist 5P12 CCL5 has been selected as lead compound for drug development , and, asFurther inspection of the CCR5 binding cavity and the extensive occupancy by 5P7 CCL5 allowed the identification of at least two new positions amenable of amino acid substitution. These substitutions have been tested by the SWISS-MODEL generation of 3D models of the CCL5 mutants that indicated a likely further enhancement in receptor occupancy and therefore a possible increase in the potency of the resulting CCL5 mutants (unpublished).The analysis presented here points to the possibility to improve CCL5 in its interaction with CCR5 by modifying residues distal to the chemokine N-terminus, either by direct receptor affinity increase or by stabilization of the chemokine hydrophobic core.i1 protein, as well as a constitutively activated CCR5 coupled to Gi1, and the 6P4 CCL5:CCR5 complex also in presence of Gi1 .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.


Deltapapillomavirus DNA, the E6 oncoprotein has an important role in cell proliferation and might be related to cancer initiation and promotion. The aim of this study was to present a standardized SYBR Green-based quantitative PCR for detection and quantification of the bovine papillomavirus 2 and 13 E6 oncogenes in urinary bladder samples from cattle. Twenty-four urinary bladders from cattle displaying tumors (n = 12) and normal bladder mucosa (n = 12) were tested by quantitative PCR. Of the 12 urinary bladders with tumors, six presented bovine papillomavirus 2 DNA concentrations ranging from 1.05 × 104 to 9.53 × 103 copies/μL, while two had bovine papillomavirus 13 DNA amplified at concentrations of 1.30 × 104 to 1.23 × 104 copies/μL. The healthy bladder mucosa samples were negative for both bovine papillomaviruses. Once the results were confirmed by conventional PCR and direct sequencing, the quantitative PCR assay developed in this study was shown to be a sensitive and specific tool for detecting and quantifying the E6 ORF of bovine papillomavirus 2 and 13 in a variety of clinical samples. Our findings of identification of bovine papillomavirus 2 and 13 DNA in urothelial tumors from cattle suffering from bovine enzootic hematuria agree with data from previous studies, representing the first detection of bovine papillomavirus 13 DNA in malignant bladder lesions of cattle from Brazil.Bovine papillomavirus types 2 and 13 can induce tumors in both the cutaneous and mucosal epithelia of cattle. These viral types are associated with the development of benign cutaneous papillomas and malignant lesions in the urinary bladders of cattle, with the latter being known as bovine enzootic hematuria. Among the viral oncoproteins encoded by  Pteridium aquilinum) ingestion, which contains mutagenic and carcinogenic principles as quercetin and ptaquiloside. These compounds trigger viral gene expression that results in cell transformation, initiating the progression to malignancy (Bovine papillomaviruses (BPVs) cause benign hyperproliferative lesions of the cutaneous and mucosal epithelia, leading to distressing animal diseases, and considerable economic losses . Bovine lignancy , 3. CasePapillomaviridae family within five different genera: Deltapapillomavirus groups BPVs types 1, 2, 13, and 14; BPVs types 3, 4, 6, 9, 10, 11, 12, 15, 17, 20, 23, 24, 26, and 28 in Xipapillomavirus; BPVs 5, 8, and 25 belong to the Epsilonpapillomavirus genus; BPVs 16, 18, and 22 are classified in Dyokappapapillomavirus; and BPV7, which is the only member of the Dyoxipapillomavirus genus. BPVs 19, 21, and 27 remain unclassified . The total DNA was extracted from fragments of tumors or normal bladder tissues weighing 25 mg by using a Qiagen DNeasy Blood and Tissue kit .To detect the BPV2 and 13 DNA, the nucleotide sequences of the moderately variable E6 gene of these viral types were selected to design two sets of specific primers using Primer Blast software. These primer pairs were able to amplify approximately 120 bp of the E6 ORF . Two pos™ 4-TOPO® vector and transformed into Escherichia coli TOP10 according to the manufacturer's instructions . The recombinant plasmids were extracted using a PureLink Quick Plasmid Miniprep Kit (Invitrogen Life Technologies). To confirm the presence of the E6 gene fragment in the plasmids, direct nucleotide sequencing was performed by using a BigDye Terminator v.3.1 Cycle Sequencing Kit  with the corresponding forward and reverse primers in a 3500 Genetic Analyzer (Applied Biosystems). The recombinant plasmids with E6 fragments were quantified using a Quant-iT dsDNA BR Assay Kit in a Qubit Fluorometer (Invitrogen Life Technologies).The E6 gene fragments of each BPV type were cloned using a pCRTo quantify the E6 genes in BPVs 2 and 13, two qPCR assays were performed using 10 μL of Platinum SYBR Green qPCR SuperMix (Invitrogen Life Technologies), 20 μM of each primer, and 0.1 μL of ROX, in a final volume of 20 μL in 7500 Fast Real-Time PCR (Applied Biosystems). The following thermal profile was used: polymerase activation at 95°C for 10 min followed by 40 amplification cycles of 10 s at 95°C and 1 min at 60°C each .6 down to 1.14 × 101 DNA copies/μL and the BPV13 plasmid containing 1.54 × 106 down to 1.54 × 101 DNA copies/μL were analyzed by conventional PCR. The same dilutions were tested in duplicate three different times to evaluate the coefficients of variation (CVs) of the qPCR. The intra- and interassay CVs for the quantification cycle (Cq) values were calculated. The LOD of the qPCR performed using SYBR Green was compared to the conventional PCR assay using the same primer sets. To test the specificity of both PCR assays, the BPV2 and 13 standard curves were run under optimal assay conditions using appropriate primers (specific BPV2 primers in BPV13 samples and specific BPV13 primers in BPV2 samples). A negative control was also included in all the assays.To determine the limit of detection (LOD) of the assay, six 10-fold serial dilutions of the BPV2 plasmid containing 1.14 × 10To screen the tissue samples, a conventional PCR was performed that targeted a common sequence of the E5 L2 open reading frames (ORFs) . PCR proR2) of 0.999 (slope = −3.488) between the Cq value and the logarithm of the BPV2 copy number (2) of 0.998 (slope = −3.698) between the Cq value and the logarithm of the BPV13 copy number (Six 10-fold serial dilutions of BPV2 and 13 plasmids were used to generate a standard curve by plotting the logarithm of the plasmid copy number against the measured Cq values . The BPVy number . The BPVy number . The eff6 down to 1.14 × 101 DNA copies/μL of BPV2 and 1.54 × 106 down to 1.54 × 101 DNA copies/μL of BPV13. A quantitative analysis of the conventional PCR identified LODs of ~114 and 154 copies of BPV2 and 13 DNA, respectively. The qPCR identified 11.4 and 15.4 copies of BPV2 and 13 DNA, respectively.The LOD of the conventional PCR and qPCR employing primer pairs designed to detect the E6 oncogene were evaluated by testing six 10-fold serial dilutions of the DNA standards, which contained 1.14 × 10The positive controls for both viruses were detected using the qPCR assays and the conventional PCRs employing the same primer pairs targeting the BPV E6 oncogene. No cross-reaction was observed when primers specific for the BPV2 E6 gene were tested against BPV13 DNA or vice versa. The reproducibility was analyzed using six 10-fold serial dilutions of a standard for BPVs 2 and 13. The BPV2 qPCR intra-assay CVs ranged from 0.065 to 1.077%, while the interassay CVs ranged from 3.81 to 9.99% . The BPVThrough PCR using the E5L2 primers, amplicons of the expected length, measuring ~250 bp, were amplified from 8 out of 12 bladder tumor samples. The direct sequencing of the resulting E5L2 products demonstrated the presence of BPV2 in six bladder tumor samples and the identification of BPV13 DNA in two samples of bladder tumors. The nucleotide sequence identity for the E5L2 fragments varied from 98.6 to 100% for BPV2 and 99.8 to 100% for BPV13.4 to 9.53 × 103 copies/μL, and two bladder samples presented BPV13 DNA at a concentration of 1.30 × 104 to 1.23 × 104 copies/μL. The 12 healthy bladder mucosa samples were negative for both BPVs. Nucleotide sequence analyses of the ~120 bp amplicons confirmed the BPV types in the amplified products.In the analyses of the clinical samples by SYBR Green qPCR as developed in this study, six out of 12 tumor samples presented BPV2 DNA at concentrations ranging from 1.05 × 10In this study, a new real-time SYBR Green-based qPCR system was developed and tested for the detection and quantification of the E6 ORF of BPV types 2 and 13.The qPCR strategy developed herein presented high sensitivity, specificity, reproducibility, and efficiency for the qualitative and quantitative detection of the E6 oncogene of BPVs 2 and 13 in clinical specimens from affected animals, despite the close phylogenetic relationship among these BPV types . Once thDeltapapillomavirus genus of the Papillomaviridae family, are involved in the benign growth of the cutaneous epithelium as well as in malignant lesions present in the mucosa of cattle. Among Delta-PVs, BPV2 is the viral type most frequently associated with the carcinogenesis of the urinary bladder of cattle who feed on bracken ferns (BPVs 2 and 13, which are classified as members of the en ferns , 12, 16.en ferns . DespiteIn Brazil, where cattle breeding is hampered in several regions due to the endemicity of BEH, previous investigations on the participation of BPV in the etiology of urinary bladder cancer confirmed the presence of BPV2 DNA through conventional specific PCR assays , 20. In Deltapapillomavirus genus, was detected in urothelial tumors from cattle from Italy, mostly accompanied by BPV2 and/or BPV13 DNA in multiple infections, being the BPV14 DNA identified in a single infection in only a tumor sample (In this study, four out of 12 malignant lesions of the urinary bladder, histologically confirmed as carcinomas, tested negative for BPV2 and BPV13 DNA in the novel qPCR assay. In a recent molecular investigation of 39 urothelial tumors from cattle suffering from BEH from southern Italy, for the presence of BPV2 and BPV13 DNA through conventional PCR techniques, five carcinomas also tested negative for both viral types . Once urr sample . Howeverr sample .To the authors, the main limitation of this study was the restricted number of bovine urothelial tumors examined by the novel qPCR system which hampered the elucidation of the frequency of naturally occurring malignant bladder tumors presenting single or multiple infections by BPVs types 2 and 13 in cattle presenting BEH from southern Brazil. However, further investigations aiming to evaluate a larger number of samples from urothelial tumors and healthy urinary bladders for the presence of BPV 2 and BPV13 DNA are needed to be performed in cattle from endemic areas for BEH in Brazil.In conclusion, the new real-time quantitative PCR assay developed to detect and quantify the E6 oncogene in BPVs 2 and 13 may be a useful molecular tool for diagnostic purposes in clinical cattle samples, allowing for epidemiological investigations on the prevalence of these BPV types in diverse geographical areas as well as their association with different clinical outcomes. This assay thus contributes to studies on the pathogenesis of BPV and the consequent development of preventive and curative strategies to control important diseases due to BPV infections that cause severe economic losses in the cattle industry.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.Ethical review and approval was not required for the animal study because the biological samples employed in the study were collected from slaughtered cattle from a private abattoir.BKA carried out most of the experiments, with the collaboration of AMDA. BKA and ML analyzed the data and wrote the manuscript. AAA and AFA procured funding and supervised the project. All authors contributed to the article and approved the submitted version.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.


Copper plays a key role in cancer metastasis, which is the most common cause of cancer death. Copper depletion treatment with tetrathiomolybdate (TM) improved disease-free survival in breast cancer patients with high risk of recurrence in a phase II clinical trial. Because the copper metallochaperone ATOX1 was recently reported to drive breast cancer cell migration and breast cancer migration is a critical factor in metastasis, we tested if ATOX1 expression levels in primary tumor tissue could predict the TM treatment outcome of breast cancer patients at high risk of recurrence. We performed ATOX1 immunohistochemical staining of breast tumor material (before TM treatment) of 47 patients enrolled in the phase II TM clinical trial and evaluated ATOX1 expression levels in relation with patient outcome after TM treatment. Our results show that higher ATOX1 levels in the tumor cell cytoplasm correlate with a trend towards better event-free survival after TM treatment for triple-negative breast cancer patients and patients at stage III of disease. In conclusion, ATOX1 may be a potential predictive biomarker for TM treatment of breast cancer patients at high risk of recurrence and should be tested in a larger cohort of patients. Copper levels are increased in serum and tumor tissue of cancer patients, and copper is required for at least three cancer hallmarks: proliferative immortality, angiogenesis and metastasis . Since fThe copper metallochaperone antioxidant protein 1 (ATOX1) is upregulated in breast, colorectal, uterus and liver tumors, while downregulated in bile duct and pancreatic tumors . ATOX1 rA recent phase II clinical trial tested the therapeutic efficacy of adjuvant tetrathiomolybdate (TM) following chemotherapy in triple-negative breast cancer (TNBC) patients, who showed no evidence of disease (NED), but were at high risk of recurrence. TNBC is characterized by its aggressive nature and lack of targeted treatment leading to a high prevalence of metastatic disease and death, respectively ,8. The pGiven that the reliance on copper trafficking in metastasis might vary across the different molecular subtypes of breast cancer and subsequently influence the impact of copper depletion as a therapeutic strategy, we sought to evaluate ATOX1 as a predictive factor for TM treatment outcome of breast cancer patients at high risk of recurrence using tumor tissue sections and follow-up data from the phase II clinical trial described above.Patients with high risk of recurrent breast cancer were enrolled onto a phase II clinical TM study NCT00195091) between 2007 and 2014. The study was approved by the Institutional Review Boards of Weill Cornell Medicine (IRB 0309006307) and Memorial Sloan Kettering Cancer Center (IRB 21-470). Additional details on the clinical trial design have been reported previously [5091 betwThe ATOX1 immunostaining of breast tumor tissue sections was performed as described previously . ShortlyLog-rank test was used to examine the relationship of ATOX1 staining with the relative probability for EFS. Survival curves were computed according to the Kaplan–Meier method in the SPSS software.ATOX1 expression was evaluated in whole breast tumor tissue from breast cancer patients enrolled in the phase II clinical trial of TM using immunohistochemical staining. Visual inspection of the stained tumor sections indicated ATOX1 expression to vary between different tissue sections from negative to strong staining intensities and to be homogeneous within each tissue section, with comparable ATOX1 staining intensity in inner tumor and tumor border. A more detailed look showed ATOX1 staining in both nucleus and cytoplasm of different cell types, including epithelial cells, fibroblasts and immune cells.We performed a semi-quantitative evaluation of ATOX1 staining intensities in the tumor cells only, considering both nucleus and cytoplasm, using intensity scores ranging from 0 to 3 representing negative, weak, intermediate and high staining intensities, respectively. In addition, we scored the percentage of tumor cells with the strongest ATOX1 staining intensity as a measure for tumor heterogeneity and found a high level of tumor homogeneity, with an average of 79% and 82% considering nuclear and cytoplasmatic ATOX1 levels, respectively. Illustrations for ATOX1 IHC staining and ATOX1 intensity scores are depicted in For analysis, we divided the scoring data into two groups: low ATOX1, which includes negative and weak staining intensities , and high ATOX1, which includes intermediate and strong staining intensities . high = 16 out of 16 (100%)) followed by Luminal (nhigh = 16 out of 18 (88%)) and TNBC subtypes (nhigh = 17 out of 23 (74%)), and we found a similar prevalence of high nuclear ATOX1 levels in the tumor tissue from patients with Luminal, HER2 and TNBC subtypes of disease (nhigh = 12 out of 18 (67%), nhigh = 4 out of 6 (67%) and nhigh = 15 out of 23 (65%), respectively) (high = 19 out of 23 (82.6%)) and IV (nhigh = 19 out of 21 (90.5%)) versus stage II (nhigh = 1 out of 3 (33.3%)) and a similar prevalence of high nuclear ATOX1 levels for patients with disease stages II, III and IV (nhigh = 2 out of 3 (66%)), nhigh = 16 out of 23 (69%) and nhigh = 13 out of 21 (62%), respectively)  A,B. Compctively) C,D.To understand if ATOX1 levels in the cancer tissue relate to TM treatment outcome, we evaluated the correlation between ATOX1 expression levels and EFS in the patients. We found that high ATOX1 levels in the tumor cell cytoplasm, but not ATOX1 in the nucleus, correlated with a trend towards better EFS when considering all breast cancer subtypes  A,B.p = 0.45) (p = 0.018) (n = 3).Deeper investigation of the correlation between ATOX1 expression levels and patient outcome after TM treatment for the different subtypes separately demonstrated that this trend of better EFS for high cytoplasmatic ATOX1 was evident for the TNBC patient cohort, with an EFS of 82% for high ATOX1 versus 67% for low ATOX1 ( = 0.45) A. Howeve= 0.018) B. There Copper depletion with TM is a promising treatment for breast cancer patients. A first phase II clinical trial for TM treatment of breast cancer patients with high risk of recurrence demonstrated an EFS of 71.4% at a median follow-up of 10.3 years . Since AATOX1 mRNA expression levels for the HER2 subtype, followed by no change for the Luminal subtype and lower levels in the Basal subtype, and a trend of increasing ATOX1 mRNA levels with higher stages of disease [Classical immunohistochemistry was used to manually score ATOX1 levels in tumor cells and the staining intensities were grouped into low (intensity scores 0 and 1) and high (intensity scores 3 and 4) ATOX1 levels. ATOX1 expression was found and evaluated for both nucleus and cytoplasm of the tumor cells. The staining results for ATOX1 in tumor tissue sections demonstrated that high ATOX1 levels for the tumor cell cytoplasm were most often found in the HER2 subtype, followed by Luminal and TNBC subtypes; in addition, high ATOX1 levels were more frequent at disease stages III and IV in comparison to stage II. These results are in agreement with our previous results of increased  disease .We found that breast cancer patients at stage III of disease with high cytoplasmatic ATOX1 levels in their primary tumor have a 65% higher EFS chance after copper depletion treatment with TM . Evaluation of the different subtypes showed that TNBC patients with high cytoplasmatic ATOX1 levels had an EFS which is 15% higher than for patients with low cytoplasmatic ATOX1 levels. For the HER2 and Luminal subtypes, no conclusions were drawn due to the low sample size. The ATOX1 correlation in TNBC suggests that, here, ATOX1-dependent processes play a crucial role in cancer progression (such as metastasis), and therefore copper depletion with TM may have a beneficial effect ,11. CoppTaken together, cytoplasmatic ATOX1 has the potential to be a predictive biomarker for TM treatment of breast cancer patients at high risk of recurrence, such that patients with high cytoplasmatic ATOX1 levels in the cancer tissue might benefit from TM treatment. We note that a larger sample size is urgently needed to come to conclusions for the HER2 and Luminal subtypes. However, the larger sample size for the TNBC subgroup implies that ATOX1 may be a predictive biomarker for TM in this cancer subtype. A larger randomized phase II clinical trial of TNBC patients is scheduled to begin in late 2021 supported by the Translational Breast Cancer Research Foundation and the NCI NExT program.


SF3B1, SRSF2, U2AF1, and ZRSR2. Interestingly, they are involved in the recognition of 3′ splice sites and exons. It has been reported that mutations in these splicing regulators result in aberrant splicing of many genes. In this review article, we first describe molecular mechanism of pre-mRNA splicing as an introduction and mainly focus on those four splicing factors to describe their mutations and their associated aberrant splicing patterns.Pre-mRNA splicing is an essential process for gene expression in higher eukaryotes, which requires a high order of accuracy. Mutations in splicing factors or regulatory elements in pre-mRNAs often result in many human diseases. Myelodysplastic syndrome (MDS) is a heterogeneous group of chronic myeloid neoplasms characterized by many symptoms and a high risk of progression to acute myeloid leukemia. Recent findings indicate that mutations in splicing factors represent a novel class of driver mutations in human cancers and affect about 50% of Myelodysplastic syndrome (MDS) patients. Somatic mutations in MDS patients are frequently found in genes  AC (underlined A is a branch point) among introns, the conserved sequence around branch point in mammals is YUNAY , which is more diverse  in MDS with the aberrant splicing mechanism caused by those mutations and an outline of MDS from a splicing point of view.SF3B1 is one of the components of the SF3B complex that stabilizes U2 snRNP binding to the branch point sequence during pre-mRNA splicing  2,29]. . 29]. SFSRSF2, originally called SC35 , is a meIt has also been demonstrated that MDS-responsible mutations in SRSF2 and U2AF1 cause expansion of R loop ,66. ExpaU2AF1 is originally identified as a component of the U2 snRNP auxiliary factor complex (U2AF) that facilitates association of U2 snRNP to the branch point sequences ,68. U2AFZRSR2 mutations cause abnormal splicing via intron retention of U12-depedent minor introns [The ZRSR2 gene is located on chromosome Xp22.2 and mutated in about 5% of MDS patients, predominantly males . Relativ introns . Humans  introns ,4. Among introns .Most recently, it was demonstrated that impaired minor intron excision by knock-out of ZRSR2 protein enhances hematopoietic stem cell self-renewal, and mutations in minor introns are suggested to be potential cancer drivers . HoweverLUC7L2. LUC7L2 is an ortholog of splicing factor LUC7 which is involved in recruitment of splicing factors. The LUC7L2 protein is assumed to be involved in the recognition of non-consensus splice donor sites in association with the U1 snRNP [In addition to four splicing factors, rare mutations in several other splicing factors were also identified. One of the mutated splicing factors is PRPF8. Studies from the yeast Prp8 protein revealed that PRPF8 protein is an essential factor for splicing and interacts with U5 snRNA to align 5′ and 3′ splice sites in the spliceosome . PRPF8 mU1 snRNP . InteresU1 snRNP . SeveralU1 snRNP . Very raU1 snRNP , which aIn this review, we introduced four major splicing factors mutated in MDS with aberrant splicing caused by mutations. Interestingly, most of the proteins described in this review are involved in 3′ splice site recognition . In addiAlthough some aberrant splicing patterns in dysregulated genes have been identified to be involved in MDS onset as described above, it is still under investigation how different mutations in different splicing factors cause different MDS phenotypes. To date, there seems to be no common gene(s) whose aberrant splicing is responsible for MDS onset caused by mutations in four main splicing factors SF3B1, SRSF2, U2AF1, and ZRSR2. It is assumed that hot spot mutations among them in SF3B1, SRSF2, and U2AF1 do not cause reduction of the encoded proteins, whereas mutations in ZRSR2 reduce functional protein amount. Splicing pattern analyses implicate that common pathways affected by mutations of those factors are epigenetics and signal transduction pathways. These points have to be addressed in future analyses. The approaches from mechanistic analyses of aberrant splicing caused by mutated splicing factors should shed light on research for therapies of MDS by identifying drug targets.


Our findings provide a novel conceptof using tunable quantum tunneling for highly sensitive broadbandphotodetection in mixed-dimensional van der Waals heterostructures.Graphene-based vander Waals heterostructures are promising buildingblocks for broadband photodetection because of the gapless natureof graphene. However, their performance is mostly limited by the inevitabletrade-off between low dark current and photocurrent generation. Here,we demonstrate a hybrid photodetection mode based on the photogatingeffect coupled with the photovoltaic effect via tunable quantum tunnelingthrough the unique graphene/Bi Variousmethods have been introduced to overcome limitations by enhancingthe light absorption of graphene-based photodetectors, including plasmonicnanostructures8 and quantum dots.9 Although these approaches have unique advantages,their photodetection is limited to a short spectral range becauseof the sharp resonant absorption.11Lightdetection over a wide spectral range, from the visible tothe mid-infrared, has a great potential for numerous photonic andoptoelectronic applications. In this scenario, graphene can providea versatile platform for broadband photodetection due to its gaplesselectronic band structure.13 In graphene-based van der Waals heterostructures, semiconductingtwo-dimensional transition-metal dichalcogenide materials are typicallyused as they exhibit superior light–matter interaction propertiesthat allow for enhanced light absorption. However, the absorptionis limited within the visible spectral range due to their band gap.14 Combining graphene with narrower band gap materialshas been proposed for broader light absorption despite the large darkcurrent.17 Among them, most of the studies using topologicalinsulators focused on the formation of graphene/topological insulatorheterostructures to induce the photogating effect, utilizing the novelproperties of topological insulators such as graphene-like hexagonalsymmetry, direct narrow band gap, and ultrafast photocurrent alongthe surface.19 As a material candidate to formvan der Waals heterostructures with graphene, topological insulatorshave advantages beyond the aforementioned properties. For example,compared to black phosphorus, which is similar to a topological insulatorin terms of band gap and electrical properties such as mobility andcarrier multiplication, the surface oxidation of black phosphorusis related to the degradation mechanism and environmental instability,20 while the surface oxidation of the topologicalinsulators serves to protect the surface states.22 Moreover, since the topological insulators are Dirac fermion materialslike graphene, a Dirac-source field-effect transistor can be realizedwith the graphene/topological insulator heterostructures.23Hybridgraphene systems combined with mixed-dimensional materialsfor high-sensitivity broadband light detection have emerged from recentdevelopments by assembling materials into van der Waals heterostructures.7 However, the photodetectionperformance using this method is highly sensitive to the size, thickness,and quality of the interlayer, which makes the fabrication processchallenging. As an alternative pathway free from introducing an interlayer,the natural oxidation layer rapidly formed on the surface of topologicalinsulators22 in the ambient environment canbe utilized as the tunneling barrier.26 In particular, the thicknessof the oxidation layer of the topological insulators is saturatedwithin a few hours after exfoliation, and the oxidation process issignificantly delayed over time so that a uniform oxidation layercan be obtained over the entire surface.22Recent studies reported the control overthe dark current in graphene-basedphotodetectors by introducing an interlayer  tunnelingbarrier with enhanced photodetectivity.2Se3 heterointerface enablesincorporation of the quantum tunneling effect into the photodetectionmechanism, as evidenced by the transition of charge carrier transportmechanisms from direct tunneling to Fowler–Nordheim (FN) tunnelingand/or thermionic emission. In our device architecture, the photogatingeffect in the graphene/Bi2Se3 heterochannelis coupled to the photovoltaic effect through the rectifying tunnelingjunction. This significantly enhances the photodetection performance,which is fundamentally different from the typical graphene-based photodetectorspreviously reported.4 Accordingly, the normalized photocurrent-to-dark current ratio (NPDR)is enhanced by around an order of magnitude via electrical tuningof tunneling resistance for detection of light covering the majorphotonic spectral region from the visible to the mid-infrared wavelengths.Our work provides a new perspective on both the tunneling dark currentsuppression and the efficient photocurrent generation for variousphotonic and optoelectronic applications.Here, we demonstrate that the naturally formed oxidation layerat the graphene/Bi2Se3 heterostructure . The Eg2 peak intensity mapping imageof the Bi2Se3 flake . On the other hand, the 2D peak intensitymapping image of graphene, as shown in 2Se3 flake .Our Dirac-sourcefield-effect transistor based on a lateral heterochanneland a vertical tunneling junction is realized by the graphene/Bicroscopy 1b and Raayer see S1a. On t2Se3 flake canbe clearly identified by the AFMimages, as shown in S2. The average thicknessesof the graphene and Bi2Se3 flakes, measuredby AFM, are ∼1.3 and 29.2 nm, respectively (see the Supporting Information for more details on theAFM)29.The graphene layer covering thesurface around the edge of Bi2Se3 (Bi: 5d and Se: 3d) takenafter 24 h from exfoliation. The observed oxidation peaks correspondingto Bi2O3 (at around 26 and 29 eV) and SeO2 (at around 59 eV) represent the existence of the oxidationlayer naturally formed on the Bi2Se3 surface.22 The intensity of the SeO2 peak ismuch lower than that of the Bi2O3 peak, indicatingthat the dominant oxidation layer formed on the Bi2Se3 surface is Bi2O3 rather than SeO2 due to the Se vacancies on the Bi2Se3 surface.32 Note that the oxidation time of 24 h after exfoliationunder ambient conditions is set to utilize the uniform oxidation layerwith the stabilized thickness. Although the formation of the nativeoxidation layer is very fast at the initial stage after exfoliation,22 its thickness is known to saturate since the oxidation process issignificantly delayed over time due to the interplay between surfaceexposure and oxygen incorporation.22 Thethickness of the oxidation layer is estimated as ∼2 nm (±0.2nm).2613 First, we investigated the current–voltage (I–V) characteristics of the graphene/Bi2Se3 heterojunction by choosing different metalelectrodes (see the 2Se3 heterointerface (definedas graphene-Bi2Se3), the I–V curves exhibit the nonlinear I–V relationship  dueto charge carrier transport through the graphene/Bi2Se3 heterointerface. The asymmetric rectifying behavior indicatesthat the tunneling junction is formed at the interface, and the tunnelingbarrier heights are asymmetric. The hysteresis effect of I–V curves arises from charge trapping atthe interface. For comparison, we also measure a reference graphenetransistor (defined as graphene&Bi2Se3),where both source and drain are applied to the graphene channel thatpartially covers the Bi2Se3 flake (see the Supporting Information for details on the measurementconfiguration with different electrodes). The I–V curves, as shown in Figure S3a, reveal the typical Ohmic behavior in the reference graphene transistor.Thevan der Waals heterostructures can enable versatile functionalitieswith higher performance than each material in the van der Waals heterostructures.2Se3, as shownin IDS–VDS curve(drain–source current IDS as afunction of drain–source voltage VDS) at gate–source voltage VGS =0 V is divided into nine steps, which are marked by Roman numeralsfrom I to IX. Each step represents the transition point, where thetransport mechanism changes and the resistance state switches to differentresistance states. The oxidation layer naturally formed on the Bi2Se3 surface is known to act as a tunneling barrierin contact with graphene.26 The tunneling resistance is closely related to thepotential barrier at the interface and the electronic density of statesin graphene and Bi2Se3. Hence, the shape deformationof the asymmetric tunneling barrier will have a great influence onthe tunneling current across the interface. The energy band alignmentsbefore equilibrium and between each step is drawn in 36 . The detailed descriptions forthe hysteretic I–V characteristicsand charge carrier trapping processes at the graphene/Bi2Se3 interface can also be found in the Supporting Information.38 Further details onthe energy band alignment are fully discussed in the Supporting Information.46To understand the mechanism of asymmetric rectifying and hystereticcharacteristics of the graphene-Bi4 On the contrary, in our device, the photogating effect is coupledto the photovoltaic effect through the rectifying tunneling junctionacross the graphene/Bi2Se3 heterointerface,which is supported by the observation of asymmetric rectifying andhysteretic I–V characteristics,as shown in 49 Some of the photogenerated carriers accumulated atthe trap states can act as an external bias voltage to shift the Fermi-levelof graphene, and the other carriers injected into graphene or Bi2Se3 will contribute to the photocurrent. On theother hand, the photovoltaic effect is driven by separating photogeneratedelectron–hole pairs through the rectifying junction, whichcan be further controlled by tuning the tunneling resistance.7 Before exploring the photogating effect, we first characterizedthe enhanced photocurrent with the photovoltaic effect, owing to therectifying tunneling junction, as shown in Figure S3. Interestingly, under the same condition of light illumination(at a wavelength of 532 nm with a laser power of 10 μW) focusedonto the same graphene/Bi2Se3 heterojunctionregion, much higher photocurrent is realized through the graphene/Bi2Se3 heterointerface , as compared tothe reference graphene transistor . This is because thephotocurrent generated in the reference graphene transistor is hinderedby the carrier recombination within the graphene channel, while thephotocurrent of the rectifying tunneling junction formed through thegraphene/Bi2Se3 heterointerface is enhancedwith the photovoltaic effect.Most graphene-based photodetectors generallyfocus on one photodetectionmechanism: photovoltaic effect, photogating effect, photo-thermoelectriceffect, and bolometric effect, due to their inherent limitations.VDS in our graphene/Bi2Se3 heterochanneldue to the strong coupling between the photogating and photovoltaiceffects. Here, two different VDS (0.5and 1.5 V) are chosen to define the high and low tunneling resistancestates. Both exhibit high photocurrents, but dark currents are obtainedto be considerably different for a proper comparison. Note that thisis based on the color plots of IPC, asshown in IPC = Ilight – Idark is the photocurrent, Ilight is the drain–source current underlight illumination, and Idark is the drain–sourcecurrent in the dark. In addition, it is found to be more effectivefor modulating the tunneling resistance by tuning positive VDS along the Bi2Se3 side,due to the lower barrier height for electrons on the graphene sidethan that toward the Bi2Se3 side, as estimatedin VDS = 0.5 V), the direct tunneling from graphene to Bi2Se3 will be substantially blocked by the tunneling barrier underthe dark condition, while the photoexcited electrons in graphene canbe easily injected into Bi2Se3 over the lowbarrier height. On the other hand, at the low tunneling resistancestate (VDS = 1.5 V), the dark current ascribed to theFN tunneling and thermionic emission will exceed the current due tophotoexcited electrons. As a result, the dark current will be obtainedto be extremely lower in the high tunneling resistance state (2Se3 heterojunction at VDS = 0.5 V (at a wavelength of 532 nm with a laser power of 100 μW).The photocurrent generation is pronounced around the heterojunctionregion especially near the edge of the graphene channel overlappingthe Bi2Se3 flake, confirming the major photocurrentgeneration originating from the heterointerface due to the built-inelectric fields applied across it.We also find that the tunnelingresistance can be significantly tuned by varying ce state 4b than tce state 4c. As shIDS as a function of VGS) of the graphene/Bi2Se3 heterointerfacein the dark and under light illumination at various wavelengths of532, 730, 1550, and 4000 nm with a light power of 10 μW areshown in VDS = 0.5 V) and VDS = 1.5V). The graphene/Bi2Se3 heterochannel operatesas a field-effect transistor, where the carrier mobility depends onthe tunneling resistance. The high (VDS = 0.5 V) and low (VDS = 1.5 V) tunnelingresistance states lead to the different average carrier mobilitiesof 36.8 and 103.2 cm2 V–1 s–1 for holes and 12.6 and 64.1 cm2 V–1 s–1 for electrons at room temperature, respectively.From the shift of VDirac, we estimatedthat the photogating effect is attributed to the trapping of photogeneratedcarriers at the graphene/Bi2Se3 interface (seethe Supporting Information for detailson the photogating effect due to the trapping of photogenerated carriers).As shown in the transfer curves, there are almost no hysteresis effectsfor the VGS sweep, thanks to the Al2O3 top passivation layer,50 implying that charge trapping at the graphene/Bi2Se3 interface only occurs during the VDS sweep. In particular, the tunneling dark current is obtained tobe quite low, as shown in Ilight/Idark). At VGS = 0 V, althoughthe photocurrents (IPC = Ilight – Idark), asshown in IPC, where Ilight = Idark, does not appearin Ilight to Idark. Unlike thephotogating effect in which one type of photogenerated carriers shouldbe captured in trap states, the photovoltaic effect requires the efficientseparation of created electron–hole pairs. Our results suggestthat the tunneling junction in the graphene/Bi2Se3 heterochannel can be utilized to couple the photogating effect withthe photovoltaic effect to enhance the optical switching ratio bytuning the tunneling resistance.The transfer curves(2Se3 is confirmed by photoresponsivity (R = IPC/Peffective) and photodetectivity . Onthe contrary, the photoresponse characteristics of the reference graphenetransistor  is almost unchangedbetween VDS = 0.5 and 1.5 V. In otherwords, the light absorption in the graphene/Bi2Se3 heterostructure does not guarantee the high photodetectivity. Thishighlights that the charge carrier transport through the graphene/Bi2Se3 tunneling junction is of great importance tocouple the photogating effect with the photovoltaic effect in thegraphene/Bi2Se3 heterochannel for highly sensitivephotodetection.Depending on the tunnelingresistance, the electrically tunable photoresponse of graphene-Biectivity 5b, , whenterface5b can beI) at VDS = 0.5 V and VGS = 0 V is plottedin Supporting Information for details on the relation between light powerand responsivity).514, 6.6 × 103, 3.8 × 103, and 1.1 × 103 W–1 (at VDS = 1.5 V) to ∼2.2 × 105, 1.1 × 105, 3.5 × 104, and 8.0 ×103 W–1 (at VDS = 0.5 V) by effectively suppressing the tunneling dark current.The idea of tuning the tunneling resistance for enhancing the photodetectivityoffers a new insight to realize the broadband photodetection by couplingthe photogating effect with the photovoltaic effect.52The dependence of photoresponsivityon light intensity  to FN tunneling and/or thermionic emission at the high biasvoltage (low tunneling resistance state), as described in 2Se3 junction.As an alternative pathway free from an artificial introduction ofan interlayer at the interface, our findings demonstrate a new perspectiveof utilizing the oxidation layer naturally formed at the graphene/Bi2Se3 interface for both dark current reduction andefficient photocurrent generation. We confirmed that the naturallyformed oxidation layer can act as a tunneling barrier. We furtherobserved the tunable tunneling resistance and offered direct evidencefor the asymmetric tunneling barrier heights using the tunneling equations.36 An additional advantage of utilizing the naturally formed oxidationlayer is that this approach is not limited by the size, thickness,and quality of the interlayer so that large-scale devices are achievableas long as Bi2Se3 is large enough, making theentire fabrication process simple. This is similar to the currentsilicon technology, where naturally formed silica plays a key role.Another novelty of our work is describing the transition of chargecarrier transport mechanisms through the graphene/Bi2Se3 interface, which is essential for coupling between photogatingand photovoltaic effects. To the best of our knowledge, the hysteresiseffect of I–V curves in graphene/Bi2Se3 heterojunction is first observed in this work,implying the trapping of charge carriers at the interface. This observationsuggests that the trap-assisted photogating effect can be inducedin the graphene/Bi2Se3 heterochannel by trappingof photogenerated carriers,49 which can be coupled with thephotovoltaic effect to effectively enhance the photocurrent owingto the rectifying tunneling junction.7In other studies, several attempts have been madeto control thedark current by utilizing the ultrathin interlayer such as Ta2Se3 interface,26 we have explored a breakthroughway to improve the photoresponse characteristics of the graphene/Bi2Se3 heterochannel by suppressing the tunnelingdark current and injecting the photogenerated carriers. We have foundthat the tunneling resistance of the graphene/Bi2Se3 heterojunction can be tuned to couple the photogating effectwith the photovoltaic effect in the graphene/Bi2Se3 heterochannel. The transition of charge carrier transportmechanisms through the graphene/Bi2Se3 interfaceis a key signature to modulate the optical switching ratio.7 The practical application to improve the device performance throughthe interface engineering  or the materialcombination , which is beyond the scope of this study, willbe a promising research direction. This study will provide usefulinformation for designing novel photodetectors for highly sensitivebroadband photodetection.Based on the asymmetric tunneling barrier formed at the graphene/Bi2Se3 into van der Waals heterostructures. The Bi2Se3 flakes were mechanically exfoliated from bulkmaterial and transferred onto the highly doped Si substrates witha 285 nm thick SiO2 layer preprocessed with solvent cleaningand O2 plasma treatment. The Bi2Se3 flake surface was naturally oxidized under ambient conditions for24 h. Owing to the rapid surface oxidation of topological insulators,22 there was no need to introducean additional interlayer such as an insulating layer grown by atomiclayer deposition (ALD) or h-BN layer before the graphene transfer.In order to form several heterostructures on each substrate at thesame time, large-area monolayer graphene grown by chemical vapor deposition,purchased from Graphenea, was wet-transferred onto the substrates.42 For the complete and smooth coverage of the graphene layer overthe entire Bi2Se3 flake, we selected the Bi2Se3 flakes with a few tens of nanometer thick,which allowed us to minimize the defects caused by the steep surfacemorphology around the flake edges. Each graphene channel was patternedto a regular shape with electron beam lithography  and reactive ion etching (Oxford Instruments PlasmaLab 80 Plus).The lateral heterochannel was defined to be the patterned grapheneribbon and transferred Bi2Se3 flake, while thevertical tunneling junction was formed in the overlapping regions.The heterojunction area was estimated to range from 50 to 100 μm2, with an average of 83.7 μm2. Ti/Au electrodesof 5/75 nm were patterned with EBL and deposited through electronbeam evaporation (MASA IM-9912) followed by a lift-off process. Themetal electrodes on the graphene channel and Bi2Se3 flakes were patterned to be orthogonal and parallel to thegraphene channel, respectively. As a passivation layer, a 10 nm thicklayer of Al2O3 grown by ALD at 150 °C was used to prevent the surface modification from theadhesion of oxygen or water molecules. After completing the devicefabrication, an optical microscope (Olympus BX60) and Micro-Raman(WITec Alpha 300 RA+) system using 532 nm continuous wave laser wereused to characterize the graphene/Bi2Se3 heterostructure.The graphene covering the Bi2Se3 flake edgewas identified by AFM Dimension Icon (Bruker). XPS was performed onthe large area Bi2Se3 flakes using a KratosAxis Ultra ESCA spectrometer with a monoenergetic Al Kα (1486.96eV) source. The pass energy was ∼20 eV, and the X-ray spotsize was ∼200 μm. Since X-rays penetrate only the topfew layers of flakes, the XPS is useful for quantitative analysisof the surface chemical composition (the outer few nanometers) regardlessof the flake thickness.Our heterojunctionphototransistor was fabricated by integrating graphene and Bi2Se3 flakes. Thephotocurrent measurements covering from the visible range to near-infraredrange were conducted in the WITec system combined with two sourcemeters.The light beams from continuous wave lasers at 532 nm (WITec focusinnovations), 730 nm (Thorlabs MCLS1), and 1550 nm (Photonetics) werefocused onto the heterojunction through an objective lens . The optical microscopy platform system allowed us to focusthe laser beam on desired positions in the sample. The diameters ofthe light spot were around 0.87, 1.18, and 2.49 μm, respectively.The photocurrent measurements in the mid-infrared range were conductedin a home-built femtosecond laser-based microscopic system combinedwith two sourcemeters. The duration and repetition rate of the incidentpulse were 230 fs and 2 kHz. The laser at 4000 nm is focused to coverthe graphene/Bi2Se3 heterojunction region bya parabolic mirror. The diameter of the light spot was ∼20μm.The sample holder was designed in a sizefitting into a fixingholder in a probe stage. After device fabrication, the device chipswere mounted onto each printed circuit board (PCB) attached to thesample holder and wire-bonded to the PCB. All the electrical measurementswere carried out in an optical microscope (WITec Alpha 300 RA+) ora home-built femtosecond laser based microscopic system with two sourcemeters(Keithley 2400 and 2401) at room temperature under ambient conditions.The gate voltage was applied to the Si substrate, while the sourceand drain voltages were applied to the metal electrodes connectedto the graphene channel or Bi


The authors apologize for the original versions of our paper unfortunately contained some incorrect representative images. We used the wrong representative images during figure assembly. The IHC images of 7dCon-Runx2, 10dCon-CD146/COL1A1 and 10dT-Del-Runx2 in Figure The authors confirm that the corrections made in this erratum do not affect the original conclusions. The authors apologize for any consequences that these errors may have caused.


Our understanding of the renin-angiotensin-system (RAS) has been revolutionized over the last 20 years with the discovery of angiotensin converting enzyme (ACE)2 and its principal product – angiotensin (Ang)-1–7) –7 1,2]. Additional convertases such as circulating furin and transmembrane protease serine (TMPRSS2) strengthen viral attachment to membranal ACE2 by modification and cleavage of the S1/S2 spike proteins Viral attachment to membranal ACE2 is followed by viral internalization together with ACE2 and ACE2 degradation, leading to depletion of membranal ACE2 Taken together, high expression of membranal ACE2 facilitates viral homing, but its loss by degradation and shedding leads to Ang-(1–7) depletion, promoting the devastating clinical manifestations of COVID-19 disease EBioMedicineACE2 transcript. It is important to consider the existence of alternatively spliced isoforms (For that reason, the study of Nikiforuk et al., published in isoforms  and to nAll authors contributed to conceptualization, writing, reviewing, editing and have read and agreed to the published version of the manuscript.The authors declare no conflict of interest.


APX2 and HSP22) and H3K4me3 following recurrent heat stress. Furthermore, CSN5A is also required for the deposition of H3K4me3 following recurrent heat stress. Thus, CSN5A plays an important role in the regulation of histone methylation and transcriptional stress memory after recurrent heat stress.In nature, plants are exposed to several environmental stresses that can be continuous or recurring. Continuous stress can be lethal, but stress after priming can increase the tolerance of a plant to better prepare for future stresses. Reports have suggested that transcription factors are involved in stress memory after recurrent stress; however, less is known about the factors that regulate the resetting of stress memory. Here, we uncovered a role for Constitutive Photomorphogenesis 5A (CSN5A) in the regulation of stress memory for resetting transcriptional memory genes (