Early detection of kidney cancer improves survival; however, low prevalence means that population-wide screening may be inefficient. Stratification of the population into risk categories could allow for the introduction of a screening programme tailored to individuals.This review will identify and compare published models that predict the risk of developing kidney cancer in the general population.A search identified primary research reporting or validating models predicting the risk of kidney cancer in Medline and EMBASE. After screening identified studies for inclusion, we extracted data onto a standardised form. The risk models were classified using the Transparent Reporting of a multivariable prediction model for Individual Prognosis Or Diagnosis (TRIPOD) guidelines and evaluated using the PROBAST assessment tool.The search identified 15 281 articles. Sixty-two satisfied the inclusion criteria; performance measures were provided for 11 models. Some models predicted the risk of prevalent undiagnosed disease and others future incident disease. Six of the models had been validated, two using external populations. The most commonly included risk factors were age, smoking status, and body mass index. Most of the models had acceptable-to-good discrimination (area under the receiver-operating curve >0.7) in development and validation. Many models also had high specificity; however, several had low sensitivity. The highest performance was seen for the models using only biomarkers to detect kidney cancer; however, these were developed and validated in small case-control studies.We identified a small number of risk models that could be used to stratify the population according to the risk of kidney cancer. Most exhibit reasonable discrimination, but a few have been validated externally in population-based studies.In this review, we looked at mathematical models predicting the likelihood of an individual developing kidney cancer. We found several suitable models, using a range of risk factors (such as age and smoking) to predict the risk for individuals. Most of the models identified require further testing in the general population to confirm their usefulness. Although 62 models were found that predicted kidney cancer risk, only a few had published performance measures or external validations. There was significant variation in study design, and the majority of studies were assessed to be at a high risk of bias. The parameters of the screening approach, such as starting age and frequency of screening, could also be tailored to the predicted level of risk for each individual. This strategy requires a model that calculates the risk of developing kidney cancer for individuals. The model should not only perform well in the population of interest, but also be easy and inexpensive to use. While several risk models predicting the development of kidney cancer have been published, it is not clear which of these, if any, might be suitable for these purposes. Models that predict the risk of undiagnosed prevalent cancer and that of future development may be of interest.We aimed to systematically identify and compare published models that predict the risk of kidney cancer in the general population and describe the range of variables included, and the performance of the models and their potential applicability to population based stratification.2We performed a systematic review following an a priori established study protocol (PROSPERO ID:CRD42018116967).We performed an electronic literature search of Medline and EMBASE in November 2018, to search for risk prediction models for kidney cancer. We included literature published from January 1980 to November 2018, with no language limits, using a combination of subject headings incorporating “renal/kidney cancer”, “risk/risk factor/chance”, and “model/prediction/score” . We then manually screened the reference list of all the papers found by the search.1Published, peer-reviewed, primary research papers2Incorporating risk factors for developing kidney cancer at the level of the individual3Providing a measure of risk using two or more factors, allowing for the identification of individuals with a higher risk of kidney cancer4Applicable to the general populationWe included studies that fulfilled all the following criteria:Studies including only highly specific groups, for example, individuals receiving dialysis, were excluded. As the focus of the review was to summarise the risk prediction models for developing cancer, studies focusing on the risk of recurrent cancer were excluded. We included models that predicted the risk of developing kidney cancer in the future as well as models that predicted undiagnosed prevalent cancer.One reviewer (H.H.) carried out the search. Three reviewers (H.H./Z.L./R.T.) screened titles and abstracts to exclude clearly irrelevant papers. To ensure consistency, a random selection of 500 citations was screened by two of the reviewers (H.H./Z.L./R.T.). Additionally, 10% of the papers were screened independently (J.U.S.).The full text was examined if a definite decision to exclude could not be made based on the title and abstract alone. One reviewer (H.H.) assessed all the full-text papers. After excluding conference abstracts, 45% of the full-text papers were also screened by another reviewer (R.T./Z.L./J.U.S./G.D.S./S.G./S.H.R.). In cases where there was disagreement, the conflict was resolved by discussion between the two researchers. If the conflict could not be resolved easily, a third reviewer (J.U.S./S.G.) was consulted. Additionally, in the early stages of the full-text screening, we discussed papers for which it was unclear whether the inclusion criteria were satisfied at consensus meetings.One reviewer (H.H.) carried out basic data extraction of all of the included studies. A standardised form was developed to record the outcome(s) of the study, type of risk factors  used, and any performance measures.More in-depth data extraction was carried out independently by two researchers (H.H./R.T./Z.L./J.U.S./S.G./S.H.R./G.D.S.) for included studies that reported performance measures for the models presented . As part of this process, the included studies were classified according to the Transparent Reporting of a multivariable prediction model for Individual Prognosis Or Diagnosis (TRIPOD) guidelines Where studies included multiple different models, for example, separate models for men and women, all were included separately. In cases where multiple versions of a model for the same population and outcome—for example, with different risk factors—were included in the study, only the best performing model was selected for data extraction. After data extraction, details of the models and their performance in development and validation populations were compared to evaluate their properties and their ability to predict the development of kidney cancer.3n = 9) were identified through citation searching and screened at the full-text stage. This process is shown in detail in After duplicates were removed, the search identified 15 281 papers. Of these, 14 613 were excluded by title and abstract screening, and a further 437 were excluded after full-text assessment. At title and abstract screening, the overall agreement between reviewers was 98.8%. At full-text screening, 4.6% of the papers screened by more than one reviewer were referred to a third reviewer to resolve the conflict. A small number of additional papers  comprise tables of relative risks, for a range of maximally adjusted risk factors. A small number were developed using classification and regression tree (CART) analysis (n = 2), or logistic regression (n = 2), or based on expert opinion (n = 1). Development of the remaining models (n = 2) is not described. Whilst the performance of these studies cannot be assessed, they may be of interest to researchers carrying out validation studies.The 54 studies that describe models predicting the presence or development of kidney cancer, but not reporting performance measures, are listed in the Supplementary material. The vast majority of these models  were developed for populations of both men and women n = 8) n = 3) Of the 11 models that predict the risk of kidney cancer and have published performance measures n = 3) n = 1) n = 1) Across these 11 models, 40 risk factors were considered for inclusion and 35 were included at least once. The most commonly included risk factors are age and smoking status, which are used in five of the 11 models. Both risk factors were included in every model for which they were considered, consistent with previously identified positive associations Conversely, previously diagnosed hypertension or type 2 diabetes were both considered as risk factors in two models but were not included. This suggests that these risk factors are not strongly predictive of kidney cancer or that they are strongly associated with other included risk factors. This conflicts with the existing literature, which suggests that hypertension and diabetes have an independent positive association with kidney cancer, although in the case of diabetes this has not been well characterised n = 3) and plasma (n = 20) biomarkers. Although some of these biomarkers could be measured using tests currently available in clinical practice (n = 8), the majority are novel markers specific to kidney cancer.None of the biomarker risk factors were included in more than one study  were developed in a case-control study n = 2) Details of the development and validation of the models are given in Supplementary Tables 1–3. The majority  also have a high risk of bias in domain 1 (population). This is due to the use of a case-control study (instead of a nested case-control or cohort study) without adjustment for sampling fractions.The results of the risk of bias and applicability assessment are shown in Three models received a low score for concerns about applicability, indicating that they are well aligned with the research question Discrimination, as measured by the area under the receiver-operating curve (AUROC) was reported for 10 of the 11 risk models. The AUROC values for these risk models are shown in The AUROC values for the five models that used biomarkers to predict the risk of prevalent kidney cancer among asymptomatic populations ranged from 0.90 to 0.98  in development The biomarker models have shown good discrimination between populations of known cases and controls. As the symptoms of kidney cancer are often nonspecific and generally occur late in the disease process, such biomarker arrays capable of identifying individuals with undiagnosed prevalent disease could be a vital resource, both as a screening test in asymptomatic individuals and as a diagnostic test within clinical practice. As the performance of risk models is affected by the population in which they are tested, however, external validation of these models in appropriate cohorts would be required first to determine their performance in these populations.n = 3149), it is difficult to evaluate, given the limited reporting of performance measures. In both development and validation, this model also relied on the accurate and consistent reporting of symptoms in primary care records. As this model was explicitly developed to predict undiagnosed prevalent disease, it would not be useful for communicating risk information to individuals with the aim of encouraging lifestyle changes. Additionally, as kidney cancer is often asymptomatic in the early stages, this model also has limited use for early detection and screening. The risk evaluation based on symptoms, however, has potential for use in primary care, where it may be a useful tool for assessing the need for or urgency of referral in symptomatic patients.One model, Shephard et al 2013 Scelo et al Two other models use easily obtainable demographic and lifestyle factors to predict the future development of kidney cancer. The two models, one for men and the other for women, developed by Usher-Smith et al in 2018 Only one model included genetic risk factors. The model by Wu et al n = 11).To our knowledge, this is the first systematic review of risk prediction models for kidney cancer. Over 60 models have been developed to predict the risk of current and future disease; published information on model performance is available only for a small proportion  are most promising for this purpose. If a population were risk stratified using the base model developed by Scelo et al The use of genetic or biomarker risk factors, present in many of the models, may increase discrimination. Addition of the biomarker KIM-1 to the base model developed by Scelo et al Author contributions: Hannah Harrison had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.Study concept and design: Harrison, Usher-Smith, Griffin, Rossi, Stewart.Acquisition of data: Harrison, Thompson, Lin, Usher-Smith, Griffin, Rossi, Stewart.Analysis and interpretation of data: Harrison, Thompson, Lin, Usher-Smith, Griffin, Rossi, Stewart.Drafting of the manuscript: Harrison.Critical revision of the manuscript for important intellectual content: Usher-Smith, Griffin, Rossi, Stewart.Statistical analysis: Harrison.Obtaining funding: Usher-Smith, Griffin.Administrative, technical, or material support: Usher-Smith, Harrison.Supervision: Usher-Smith, Griffin.Other: None.Financial disclosures: Hannah Harrison certifies that all conflicts of interest, including specific financial interests and relationships and affiliations relevant to the subject matter or materials discussed in the manuscript , are the following: G.D. Stewart has received educational grants from Pfizer, AstraZeneca, and Intuitive Surgical; consultancy fees from Pfizer, Merck, EUSA Pharma, and CMR Surgical; travel expenses from Pfizer; and speaker fees from Pfizer. All other authors have no financial disclosures or conflicts of interest with respect to the research, authorship, and/or publication of this article.Funding/Support and role of the sponsor: This report is independent research supported by the National Institute for Health Research NIHR Systematic Review Fellowship, RM-SR-2017-09-009. Juliet A. Usher-Smith was funded by a Cancer Research UK Prevention Fellowship (C55650/A21464). Sabrina H. Rossi is funded by a CRUK Clinical PhD Fellowship. The University of Cambridge has received salary support in respect of Simon J. Griffin from the NHS in the East of England through the Clinical Academic Reserve. Funding was to support salary/studentships of researchers only. The views expressed in this publication are those of the author(s) and not necessarily those of the NHS, the National Institute for Health Research, or the Department of Health and Social Care.Acknowledgements: We would like to thank Isla Kuhn for her help in developing our search strategy. We thank Zhirong Yang and Mila Petrova of their assistance in screening several non-English full texts. We would also like to thank Philp Dondi and Phil Alsop, members of our PPI group, for their time and feedback on this review.


Background: Extensive studies related to vascular calcification (VC) were conducted in recent years. However, no bibliometric analysis has systematically investigated this topic. Our study aimed to determine the hotspots and frontiers of VC research in the past decade and provide a reference for future scientific research directions and decision-making in the VC field.Methods: VC studies were acquired from the Web of Science Core Collection. Bibliometric and visual analyses were performed using CiteSpace, VOSviewer, and Microsoft Excel software.Results: A total of 8,238 English articles on VC research published in 2011–2020 were obtained. In the past decade, annual publications and citations showed a significant growth trend, especially in 2018–2020. The most productive country, institution, journal and author are the United States, the University of California System, PLOS ONE, and Budoff MJ, respectively. The most frequently cited country, journal, and author are the United States, Journal of the American College of Cardiology, and Floege J, respectively. “Vascular calcification,” “atherosclerosis,” “chronic kidney disease,” and “cardiovascular disease” are the primary keywords. The burst keywords “revascularization,” “calciprotein particle,” “microRNA,” and “microcalcification” are speculated to be the research frontiers.Conclusion: The main research hotspots in the VC field are the molecular mechanisms and prognosis of VC in patients with chronic kidney disease or cardiovascular disease. In addition, endovascular therapy and the development of new drugs targeting signal pathways for VC will become the focus of future research. Moreover, non-coding RNAs related to the diagnosis and treatment of VC are great research prospects. Vascular calcification (VC) is defined as the abnormal deposition of calcium and phosphorus minerals in the arterial intima or media that can lead to reduced vascular wall compliance . VC is aVC was previously thought to involve the passive deposition of calcium phosphate crystals within the vessel wall. However, since the 1990s, extensive studies have shown that VC is an active and regulated process similar to bone development . Recent Continuous studies of VC mechanisms have made great contributions to the exploration of the best treatment to improve patient prognosis. However, no efficient therapy is approved for the treatment of VC until now, although various treatment strategies targeting pathological conditions  are available. These interventions include phosphate binders, sodium thiosulphate, calcimimetics, vitamin K, vitamin D, myoinositol, denosumab, and tissue-nonspecific alkaline phosphatase inhibitors . SeveralSeveral methods can be used to conduct a quantitative analysis of the literature, such as a traditional review, a main path analysis , and bibVOSviewer is a bibliometric software used to construct relationship networks and visualize data. Its outstanding feature is that it has strong graphics capabilities and is suitable for processing large-scale data . Using bRelevant literatures were obtained from the SCI-E database of the WoSCC on February 1, 2021. The search formula was “TS = .” The search results were confined by language (English), literature type (article), and publication year (2011–2020). Ultimately, 8,238 records were identified.CiteSpace 5.7. R4, VOSviewer 1.6.16, and Microsoft Excel 2019 were used to analyze and visualize the literature on VC research.The visualization map created by CiteSpace consists of nodes and lines. The nodes in the graph represent the node types, such as country, institution, author, keyword, and reference. The size of each node stands for the frequency, and the connections between nodes represent cooperation, co-occurrence, or co-citation relationships. Node size is proportional to its frequency by type. The number of lines represents the degree of the connection between the nodes. Betweenness centrality is applied to quantify the importance of a node’s position in the network. The higher the betweenness centrality, the greater the number of connections in the network passing through the node. Nodes with a betweenness centrality greater than 0.1 are often marked with purple circles . In our VOSviewer can be used to construct a scientific knowledge network and display the structure, evolution, and cooperation of the research field . In thisMicrosoft Excel was used to construct the tables and demonstrate the annual national trends in publications and citations.A total of 8,238 English articles related to VC were included. There were 135,220 and 16.41 total and mean citations, respectively. The H-index count was 117. The H-index represents H papers published by the journal/author/country, each of which was cited at least H times. It can be used to evaluate the scientific impact of the journal, author, or country.2 = 0.9949) between the year and the citations. The above results demonstrate that VC has received widespread attention from global scholars and that extensive research was conducted recently.From 2011 to 2020, the publications in the VC field generally showed an upward trend except for a slight decline in 2014 and 2018 . A totalAll literatures were distributed among 104 countries and 7,677 institutions. The United States had the most publications, with 2,555 (31.01%) articles, followed by China , Germany [781 (9.48%)], Japan [729 (8.85%)], and Italy [536 (6.51%)] . As showAs shown in A total of 37,697 authors contributed to VC research. Budoff MJ, the most productive author from the United States, published 68 articles, followed by Wang Y (63), De Jone PA (54), and Massy ZA (53). The top three most published authors were Floege J , Pibarot P , and Budoff MJ . The above authors’ institution distribution showed that most institutions originated from North America and Europe . As showPLOS ONE (IF2.74), Atherosclerosis (IF3.919), and Arteriosclerosis Thrombosis and Vascular Biology (IF6.604). In addition, the Journal of the American College of Cardiology (IF20.589) had the greatest number of citations, followed by Arteriosclerosis Thrombosis and Vascular Biology (IF6.604) and PLOS ONE (IF2.74) (Circulation (IF23.609), Journal of the American College of Cardiology (IF20.589), and Kidney International (IF8.52)  and journal quartile were obtained from Journal Citation Reports 2019. The top three prolific journals were (IF2.74) . The top(IF8.52) . Most joAs shown in As shown in in vitro experiments was performed to describe the molecular mechanism of VC. According to the density map of keywords, “vascular calcification,” “chronic kidney disease,” “hemodialysis,” “coronary artery calcification,” “mortality,” “vascular smooth muscle cells,” and “expression” had higher weight  had the highest IF. Although PLOS ONE  and Atherosclerosis  were the top two productive journals, their IF values were less than 5. This indicates that improving research quality while increasing output may contribute to enhancing their academic influence. Among the most commonly cited journals, the New England Journal of Medicine , Circulation , and Journal of the American College of Cardiology  had IF values higher than 20. This suggests that the three aforementioned journals published high-quality research with convincing and mature outcomes.Journal Citation Reports 2019 was used to obtain the impact factor (IF) and quartile (Q) of a journal category . The JouThrough the potent combination of top keywords and literature, we attribute the research hotspots as follows: 1) The mechanism of VC in CKD or dialysis patients, including inflammation , VSMC phBased on the analysis of burst keywords, we speculated that endovascular intervention measures for VC will become an emerging academic trend in VC research. These measures included minimally invasive transcatheter valve replacement for aortic valve or heart valve stenosis , percutaTo date, this study is the first bibliometric analysis of hotspots and dynamic frontiers of VC research in the past decade. In addition, three tools were applied to perform the survey simultaneously, enabling our research results to be more accurate and objective. However, the present study has some limitations. First, all data were extracted only from the WoSCC, and we did not search more databases, such as Google Scholar or PubMed. However, the WoSCC is an authoritative, comprehensive, and multidisciplinary core journal citation index database. Second, only English articles were included, which may have decreased the number of retrieved articles. And finally, the amount of literature related to VC is huge. With the rapid updating of hot topics and research frontiers of VC, we may have missed some research hotspots.PLOS ONE, and Budoff MJ, respectively. The most commonly cited country, journal, and author are the United States, Journal of the American College of Cardiology, and Floege J, respectively. The document with the highest number of citations is “Two-year outcomes after transcatheter or surgical aortic valve replacement” by Kodali SK in 2012. The main research foci in the VC field are the molecular mechanism of VC and the prognosis of VC in patients with CKD or cardiovascular disease. In addition, endovascular therapy and the development of new drugs targeting signal pathways for VC will become the foci of future research. Moreover, ncRNAs related to the diagnosis and treatment of VC have great research potential. In general, this study systematically analyzed the literatures on VC and reported the research results of the past decade in multiple dimensions, which can lay the foundation for the in-depth development and research of VC worldwide. Our results revealed the authors and institutions that can cooperate and provide reference for future research directions and scientific decision-making.Extensive studies related to VC have been conducted in the past decade, and the number of retrieved articles demonstrated an obvious growth trend, especially from 2019 to 2020. The most productive country, institution, journal, and author are the United States, the University of California System,


Gemella among allergic infants, and Lactobacillus and Bacteroides among healthy infants. Although infants with allergies and healthy infants demonstrate microbiota with similar α-diversity, some differences in β-diversity and bacterial species abundance can be seen, which may depend on the phenotype of the allergy. For some organisms, their abundance in skin and feces samples may be correlated, and these correlations might serve as indicators of the host’s allergic state. The gut microbiota in patients with food allergy, and the skin microbiota in atopic dermatitis patients differ from those of healthy people. We hypothesize that relationships may exist between gut and skin microbiota in patients with allergies. The aim of this study was to determine the possible relationship between gut and skin microbiota in patients with allergies, hence simultaneous analysis of the two compartments of microbiota was performed in infants with and without allergic symptoms. Fifty-nine infants with food allergy and/or atopic dermatitis and 28 healthy children were enrolled in the study. The skin and gut microbiota were evaluated using 16S rRNA gene amplicon sequencing. No significant differences in the α-diversity of dermal or fecal microbiota were observed between allergic and non-allergic infants; however, a significant relationship was found between bacterial community structure and allergy phenotypes, especially in the fecal samples. Certain clinical conditions were associated with characteristic bacterial taxa in the skin and gut microbiota. Positive correlations were found between skin and fecal samples in the abundance of  Although the cause of the growing prevalence of allergic diseases remains unclear, the Old Friends Hypothesis  has recently been proposed . The hypThe composition of the human microbiome is not only characteristic of each individual, but also evolves over time ,4,5,6,7.Similarly, the skin microbiota evolves over the years according to changes in skin structure and function , becominThe gut and the skin are crucial barriers through which environmental factors interact with the human body; the two compartments are also massively colonized with distinct microbial communities. Furthermore, they constitute complex immune organs that are fully integrated into the overall immune system. Proper skin and gut functioning is essential for homeostasis, and decreases the risk of allergy. Hence, exposure to the many environmental factors that shape the microbiome of the gut and skin epithelium can alter the epithelial surfaces and thus drive the type 2 immune response, which is known to underlay most, if not all, atopic diseases. Many studies indicate the existence of a gut–skin axis, which is modulated by microbiota and its metabolites ,17,18. Generally, the first symptoms of allergy involve the skin and/or the gastrointestinal tract , manifesThe bacterial community structure has also been found to differ between IgE-mediated and non-IgE-mediated FA children , and difStaphylococcus aureus colonizing atopic skin promoted inflammation in children aged one to three years, while commensal S. epidermidis strains might mitigate this effect [Staphylococcus (particularly S. aureus) was greater in children with AD, and that microbial diversity increased over the course of treatment [S. aureus, it has also been proven that exacerbations of skin lesions are connected to a reduction in skin microbiome diversity, while improvement is not directly related to a reduction in S. aureus, but rather to an increase in bacterial diversity [Staphylococcus richness in the skin microbiome during early infancy might be a prognostic factor of AD development before 12 months of age, and that colonization by commensal Staphylococci might protect against eczema [Similarly to the gut microbiome, the skin microbiota has also been found to differ between patients with AD and those without , and oths effect . Similarreatment . Interesiversity . In addiiversity ,31,32,33Current evidence suggests that while the gut microbiota may demonstrate a reduction in microbial diversity before AD onset, no such relationship has been noted for the skin microbiota ,23. FurtTo gain a more holistic understanding of how the microbiome of the human body interacts with the immune system, and how it can influence the development of FA and/or AD, it is necessary to study the skin and gut together as part of one combined study. The present study is the first such study to compare the microbiota of two compartments, i.e., gut and skin, in young infants with early onset of FA and/or AD. Indeed, of all the studies regarding the relationship between microbiota and allergy published to date, only one  has analThe present study characterizes the alpha and beta diversity of the fecal, i.e., gut, and skin bacterial communities from infants with FA and/or AD and compares them with those of healthy control subjects. We hypothesize that: Hypothesis 1.Gut and skin alpha diversity differ between children with an allergy (FA and/or AD) and those without;Hypothesis 2.The communities within a single compartment differ according to the clinical status of the child;Hypothesis 3.OTUs characteristic for each compartment-clinical status communities exist;Hypothesis 4.Relationships exist between the gut and skin microbiota, and these differ with regard to clinical status.To confirm these hypotheses, V3-V4 16S rRNA gene libraries were generated and sequenced, and the resulting sequences were analyzed bioinformatically. This is an observational pilot study, constituting part of Stage I of a prospective study.Participants were recruited from 1 February 2018 until 30 January 2019 among infants hospitalized in the Department of Pediatrics, Allergology and Gastroenterology, Collegium Medicum, Nicolaus Copernicus University, Poland and from Gastrological, Allergological and Nephrological Outpatient Clinics in Bydgoszcz, Poland. Enrollment was proposed to parents of 245 infants under six months of age. Informed consent was obtained from 203 of them. Out of these patients, 104 did not fulfill the inclusion criteria, or were excluded , and theFinally, 87 infants were enrolled, 59 of them with suspicion of allergies  and 28 healthy infants, comprising the control group. The allergic group comprised 38 patients with symptoms of FA and AD (ADFA group), 16 with symptoms of FA (FA group) and 5 with AD (AD group). The characteristics of the study and control group are given in The participants completed the modified ISAAC questionnaire . It compThe diagnosis of FA was based on history, physical examination, COMISS evaluation, specific IgE in the blood, elimination and oral provocation tests.COMISS (Cow’s milk-related symptoms score) was assessed based on the standard questionnaire . Scores -history of skin rash affecting the flexures , cheeks, neck, around eyes or outer surfaces of limbs;-history of atopic diseases in a first-degree relative;-history of generally dry skin;-visible eczema involving cheeks, forehead, flexures and extensor/outer surfaces of limbs.Atopic dermatitis (AD) was diagnosed based on the UK Working Party’s criteria with modifications necessary for infants ,41, i.e.-AD without FA;-AD with FA, i.e., eczema despite emollients; eczema and other typical symptoms of FA; significant improvement after elimination diet and deterioration after reintroduction of presumed allergen into the diet.AD severity was assessed based on SCORing Atopic Dermatitis (SCORAD) scale at study entry . AD patiFecal and skin samples were collected once, during the visit at the time of study enrollment. Sterile, disposable equipment was used throughout the sampling procedure. Freshly evacuated feces in diapers were gathered into collection tubes and frozen for transport to the laboratory. Skin swabs were collected from non-lesional, healthy looking skin on the cheeks; in cases where the cheeks were atopic, sampling was performed from any other healthy skin region excluding the diaper area, usually from the dorsal surface of the forearm. All of the samples were stored at −80 °C until further processing.Dermatophagoides pteronyssinus and D. farinae, cat dander, dog dander, birch, hazel, and grass pollen, olive and Alternaria spores) were tested.Serum IgE was quantified with Polycheck test  according to the manufacturer’s protocol. The detection limit was assessed as 0.35 kU/l. The patient was regarded as sensitized if specific IgE (sIgE) concentration exceeded the detection limit. Main food allergens  and inhalant ones  combined with flocculation and silica columns. V3-V4 bacterial 16S rRNA gene fragments were amplified and converted into Illumina sequencing libraries as previously described . The libp-value < 0.05 was considered significant. The sequencing reads were processed as described earlier . Brieflyp-values were corrected for multiple comparisons using Benjamini-Hochberg FDR (p.adjust); a significance level of 0.05 was used. Clinical categorical data were analyzed using two-tailed Fisher’s exact test in R (fisher.test); again, a significance level of 0.05 was used.Differences in taxa abundance were assessed with ANOVA (aov function in R); the normality of data was checked with the Shapiro–Wilk test (shapiro.test) and homogeneity of variance by Levene’s test (levene.test of the lawstat packege). Where the assumptions were not fulfilled, the Kruskal–Wallis test was used . All methods were carried out in accordance with relevant guidelines and regulations. The study protocol was approved by the local Ethics Committees of the Institutional Review Board of CM Bydgoszcz NCU Torun, Poland (765/2017). Informed written consent was obtained from the parents of all participants prior to enrollment.Sequence data generated during this project are available in the SRA database under BioProject no. PRJNA657878.No significant differences in diversity (Shannon’s H’), species richness (observed number of OTUs) and evenness (Shannon’s E) were found between controls and the participants with either AD, ADFA or FA, regardless of the compartment (skin and gut). Similarly, no differences were found between the group of allergic patients and those without an allergy. However, the diversity of skin microbiota tended to be lower in AD patients . No corrp = 0.003; p = 0.04) and between ADFA and control (p = 0.05). Significant differences were observed between the control, AD, ADFA and FA groups regarding fecal sample community structure , significant differences became apparent when age (treated as continuous variable) was added to the overall model (p = 0.013). Regarding individual group comparisons, significant differences in the skin microbiome were found between AD and FA (p = 0.014), AD and control (p = 0.01), as well as AD and ADFA (p = 0.02)  B. Any taxa whose abundance differed significantly between groups were regarded as characteristic for the group where the given taxon was more highly represented .p = 0.04 and p = 0.004, respectively); however, the reverse was true for the order Lactobacillales (p = 0.045), family Streptococcaceae (p = 0.045), genus Streptococcus (p = 0.044), and OTU13 belonging to genus Bradyrhizobium (p = 0.019).The members of the Xanthomonadales and Xanthomonadaceae were more abundant in the skin communities of all allergic (AD + ADFA + FA) patients, than in healthy children (p = 0.03), class Bacteroidia (p = 0.03), order Bacteroidales (p = 0.046), family Bacteroidaceae (p = 0.043) and genus Bacteroides (p = 0.043). At the OTU level, OTU35, affiliated with Parabacteroides, and OTU21, belonging to Bacteroides, were more abundant in allergic children , while OTU94 , OTU68  and OTU30  were more common in the control group. In the gut of allergic children, the following taxa were more abundant: phylum Bacteroidetes (p = 0.02). Sequences affiliated with Streptococcaceae were most frequent in libraries derived from skin samples of healthy children, but were absent from patients with skin symptoms , the same applied to genus Streptococcus (p = 0.039). At the OTU level, only low abundance sequences  were found to be characteristic of a particular clinical picture. Comparing all four groups allowed the identification of taxa specific for a particular clinical picture. The highest abundance of phylum Proteobacteria was found in skin samples of AD patients; lower levels were observed in ADFA and the lowest in the FA and control groups (p = 0.043); the same was observed to orders Bacteroidales and Xanthomonadales , families Xanthomonadaceae and Bacteroidaceae  and genera Stenotrophomonas and Bacteroides . In contrast, the abundance of genus Enterobacter was higher in children with no intestinal symptoms (AD and control) but lower in those with a food allergy . Differences in OTU abundance  and the families Xanthomonadaceae and Corynebacteriaceae . In addition, in the feces-derived libraries, class Bacteroidia (p = 0.007), order Bacteroidales (p = 0.004) and family Bacteroidaceae were found to be more common in AD + ADFA than in healthy children. To identify the taxa potentially involved in AD and FA pathogenesis, the results for the control group were compared with those from all patients with symptoms either on the skin (AD + ADFA) or in the gut (FA + ADFA) . Librarip = 0.016) and family Xanthomonadaceae (p = 0.016) than those from the feces samples. Differences at the OTU level were also observed: in the skin samples, OTU2 and OTU13 were more abundant among control patients than the AD and FA groups, respectively. In the feces samples, OTUs 11, 35 and 41 were more abundant in the AD group than controls, while only OTU21 was more abundant in FA patients than the controls (Libraries from the skin samples of children with FA (FA + ADFA) were more abundant in bacteria from the order Xanthomonadales . Similarly, positive correlations were observed for the genus Gemella for all allergic children (AD + ADFA + FA) , and for the genera Lactobacillus and Bacteroides in the control group . No negative correlations were observed between the skin and feces libraries.Positive correlations were observed between the skin and gut libraries for the genus Streptococcus) and OTU31  were correlated in both the allergic and control groups. Interestingly, no negative correlations were observed. In allergic subjects, the correlated OTUs were affiliated with Gram-positive phyla (Firmicutes and Actinobacteria), while those correlated in healthy subjects included the Gram-negative taxa Haemophilus (Gammaproteobacteria) and Bacteroides (Bacteroidetes). The abundance of 13 OTUs belonging to 11 genera was positively correlated between respective skin and feces samples . Among tWe examined the diversity and community structure of both skin and gut (feces) microbiota in young infants with allergies symptoms, using 16S rRNA gene amplicon sequencing. In addition to being the first of its type to be performed on Polish patients, the study has four key strengths. Firstly, the gut and skin microbiota were examined simultaneously and compared. Secondly, a novel analysis was performed on the skin microbiota in infants with FA. Thirdly, the study group was characterized by a range of clinical pictures, which allowed the differences in the microbiotas to be highlighted. Finally, unlike most previous studies, the participants were under six months of age; this allows the relationship between the microbiota and the development of allergy to be assessed in an early stage of life.The bacterial communities found in human gut and skin vary considerably between individuals and with regard to developmental stages, as do views on their relationship with allergy development ,16,22,26However, similarly to other studies ,52,53,54We observed a significant correlation between fecal bacterial community structure and allergy type, even though the communities varied greatly within one group. This result is similar to those obtained previously ,12. InteBacteroides and Parabacteroides) in the libraries derived from feces, was related to certain types of allergy. This relationship might be driven by the production of the epithelial barrier-breaching agent phenol/p-cresol, which is known to be produced by the members of the Bacteroidaceae [Some of the taxa whose abundance was found to differ between allergic and healthy infants have previously been found to be related to allergies, although the direction of these relations varied. We found that the abundance of Bacteroidales and their member taxa, down to the level of genus  were more prominent in those of healthy subjects [It seems likely that the gut microbiome influences the health status of skin ,67,68,69Our observations suggest that gut dysbiosis might be involved in the development of atopic skin inflammation, even though different organisms were differentially abundant in feces and skin libraries. Similar observations have been reported previously ,54,66,69Gemella, Lactobacillus, Bacteroides) in the skin samples with those in the fecal samples. This observation, made for the first time, supports the skin–gut axis hypothesis [In spite of the great variability in bacterial communities observed in our samples, we found certain bacterial taxa to be characteristic of particular clinical conditions, and identified a positive correlation between the abundance of certain taxa . They can also lead to the selection of specific probiotics for treating allergic conditions, either applied to the skin or applied enterally; our analysis suggests that Although the gut and skin microbiota of infants with early symptoms of allergy demonstrate similar α-diversity to those of healthy infants, some differences in β-diversity and the abundance of some bacteria can be seen, which may depend on the phenotype of the allergy. Moreover, the gut and the skin microbiota appear to be related to one another; however, this relationship may differ between patients with allergies and those without. These correlations might serve as indicators of the allergic state of the host.


Objective: Adolescents and young adults are susceptible to high-risk behaviors such as self-harm and suicide. However, the impact of childhood maltreatment on suicide attempts in adolescents and young adults with first episode of depression remains unclear. This study examined the association between suicide attempts and childhood maltreatment among adolescents and young adults with first depressive episodes.Methods: A total of 181 adolescents and young adults with first depressive episodes were included. The Child Trauma Questionnaire (CTQ), Beck Anxiety Inventory (BAI), and Patient Health Questionnaire-2 (PHQ-2) were used to assess childhood maltreatment and the severity of anxiety and depressive symptoms, respectively. The suicide item in the MINI-International Neuropsychiatric Interview (M.I.N.I.) 5.0 was used to assess the suicide attempts. Logistic regression analyses were used to explore the associated factors of suicide attempts.Results: The prevalence of SA in the total sample was 31.5% (95% CI = 24.9–38.1%). Multivariate logistic regression analyses revealed that the diagnosis of bipolar disorder , smoking , anxiety symptoms , and childhood maltreatment  were potential associated factors of SA. In addition, anxiety symptoms had a mediating effect on the relationship between childhood maltreatment and SA.Conclusion: Adolescents and young adults with first depressive episodes and having experiences of childhood maltreatment are at a high risk of suicide. The severity of anxiety symptoms may mediate the relation between childhood maltreatment and suicide attempts in this group of patients. Suicide attempt (SA), one of the major suicidal behaviors, is defined as an act of self-injuring with the intention of ending one's own life . Around Mental disorders, such as major depressive disorder (MDD) and bipolar disorder (BP), could increase the risk of suicide behaviors in adolescents and young adults –7. A metApart from the negative outcomes of physical injury and the increased risk of completed suicide, SA could worsen patients' general well-being and increase their utilization of health services and the economic burden . UnderstChildhood maltreatment, including abuse  and neglect , often has negative and long-term adverse effects on individuals' mental health –18. IncrConsidering that childhood maltreatment may lead to SA among patients with MDD and BP and suicide behaviors are most likely presented in their first depressive episode , 25, we In addition, studies among healthy adults suggested that negative emotion, such as depression and anxiety, may mediate the association between childhood maltreatment and suicide behaviors –34. SuchThis cross-sectional study is part of Youth Depression Cohort (XiangyYa) study (YDC-XY), which was conducted from January 1, 2018, to December 31, 2019, at the outpatient department of the Second Xiangya Hospital of Central South University, China. Patients who visited the hospital because of their first depressive episode were consecutively recruited if they fulfilled the following criteria: (1) aged between 14 and 24, (2) experiencing their first depressive episode, (3) diagnosed with MDD or BP according to the Diagnostic and Statistical Manual of Mental Disorders-V (DSM-V), and (4) received no psychiatric treatment (including antipsychotic drugs and psychotherapy) in the past 3 months. The exclusion criteria were as follows: (1) comorbid with neurological conditions or substance dependence and (2) participating in other clinical trials.Sociodemographic and clinical data were collected, including age, gender, education year, marital status, nationality, occupation, history of smoking, drinking, drug use, family history of psychiatric disorders and parents' marital status. Drug use history was assessed using a dichotomous item: “In your lifetime, have you ever used any of the following drugs ?.” Smoking and drinking history was assessed using two dichotomous items: “Have you ever smoked?” and “Have you ever consumed alcohol?.” SA was assessed using the item of M.I.N. I: “In the past month did you attempt suicide?.”The Child Trauma Questionnaire (CTQ) , a 28-itThe Beck Anxiety Inventory (BAI)  was usedThe Chinese version of the Patient Health Questionnaire-9 has been widely used in research and clinical practice to screen depressive symptoms . The PatThe continuous sampling method was applied. All patients who were first time visit the psychiatric outpatient services because of depression invited to participate in the survey. After obtaining written informed consent from the participants, a face-to-face interview was conducted to reconfirm the psychiatric diagnosis by a senior psychiatrist using a structured diagnostic tool, the M.I.N.I according to the DSM-V. Then all participants were asked complete all measurements mentioned above. According to patients' response to the item “In the past month did you attempt suicide?” in M.I.N.I, patients were recognized as having “suicide attempts” if they answer “yes” and as having “no suicide attempts” if “no.”t-tests and chi-square tests were used to compare the socio-demographic and clinical data between patients with and without SA, as appropriate. Multiple logistic regressions were performed to examine the independent variables associated with SA. Variables with p < 0.10 in univariate analyses were entered into the regression model as independent variables. In addition, the association between the total score of the BAI and CTQ was tested using Pearson's correlation coefficient. Mediation analysis was conducted using the PROCESS v3. 3 to examine the extent to which potential mediators may explain the relationship between childhood maltreatment and SA. Potential mediators were selected based on the results of the multiple logistic regression. The significance level was set as p < 0.05 (two sides).The SPSS 24.0 software was used for all statistical analyses. The two-independent sample This study was approved by the ethics committee of The Second Xiangya Hospital of Central South University. All patients signed written informed consent.n = 57) reported SA. The sociodemographic and clinical data are shown in A total of 181 adolescents and young adults with first depressive episodes met the eligibility criteria and were included. Among them, 31.5% (95% CI = 24.9–38.1%) (p = 0.002), drink , smoke , and present with more severe anxiety symptoms  than those without. Compared to patients without SA, patients with SA experienced significantly more severe emotional abuse , physical abuse , emotional neglect , physical neglect , and overall childhood maltreatment . There was no significant difference in the PHQ-2 total score between patients with and without SA   .r = 0.201, p < 0.001).A multiple logistic regression model revealed that the diagnosis of BP , smoking , severity of anxiety symptoms , and childhood maltreatment  were significantly and positively associated with SA . There wThe mediation analysis revealed that anxiety symptoms mediated the relationship between childhood maltreatment and SA  and explained 24.40% of the total effect , Table 3This study found that 31.5% of treatment-naïve adolescents and young adults with first depressive episodes had SA, which was higher than one previous study about the first-episode and drug naïve patients with MDD with a prevalence of 19.9% . SeveralOne of the important findings of this study is that patients with SA experienced more childhood maltreatment than those without SA. Previous studies also found a similar association between childhood maltreatment and SA among general adolescents and adults , 46. PatAnother important finding of this study is that anxiety symptoms were found to mediate the relationship between childhood maltreatment and SA among adolescents and young adults with first depressive episodes, which is consistent with previous studies . These rFurthermore, this study found that young patients with BP were more likely to have SA than those with MDD, which is consistent with previous findings , 60. ForThe strengths of the present study include the large sample size of treatment-naïve adolescents and young adults with first depressive episodes. In addition, the study examined the association between SA and childhood maltreatment among adolescents and young adults with MDD and BP, and discussed the association between childhood maltreatment and suicide behaviors among patients with depression. However, several methodological limitations should be considered. First, as a cross-sectional study, a causal relationship between SA and associated factors could not be generated. Second, some potential factors associated with SA, such as residential type and socioeconomic state , 64, werThis study found that treatment-naive adolescents and young adults with first depressive episodes were at a high risk of SA. Patients with SA were more likely to have experiences of childhood maltreatment and present more severe anxiety symptoms than those without SA. Anxiety symptoms may mediate the relationship between childhood maltreatment and SAs. Considering the severe negative outcomes of SA, it is essential to develop prevention strategies for SA among depressed adolescents and young adults. Alleviating anxiety may be an effective way to reduce the risk of suicide in this group of patients.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding authors.The studies involving human participants were reviewed and approved by the Ethics Committee of the Second Xiangya Hospital of Central South University. Written informed consent to participate in this study was provided by the participants' legal guardian/next of kin.HC, SZ, and JZ: conceived and designed the study. PL, JL, XCh, HG, CL, and XL: participated in the acquisition of data. HC: analyzed the data and drafted the manuscript. WL, XCa, JZ, and XW: revised the manuscript. All authors read and approved the final manuscript.This study was supported in part by grants from the National Natural Science Foundation of China , the Natural Science Foundation of Hunan , the Health Committee of Hunan , the Hunan Province Innovation Province Construction Project , and Clinical medical technology innovation guidance project of Hunan .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. 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Pachyphytum cesium (Crassulaceae), an endemic species from central Mexico threatened by human activities and climate change. To increase our knowledge of the species, we analyze the diversity and genetic structure of all known populations of P. caesium to change their current genetic status and propose conservation strategies. The results indicate that this species presents low to moderate levels of genetic diversity and incipiently structured populations due to low genetic flow. We found that genetic parameters are essential to the conservation strategies and status vulnerability reclassification. Based on our results, we propose reclassifying the conservation status of the species in danger of extinction; hence a series of conservation strategies are provided to prevent its disappearance.Species decline has accelerated in recent decades, with rare species often being the first to go extinct, mainly due to low population sizes. This study worked with Pachyphytum caesium is an endemic species with extremely small populations in central Mexico. This work analyzes the diversity and genetic structure of Pachyphytum cesium (Crassulaceae) to change the conservation status and propose conservation strategies. Twelve dominant ISSR markers were used to describe the genetic diversity of all known populations. Additionally, we carried out two simulations to explore how the loss of individuals or the local populations extinction affect the genetics parameters of the species. The average results indicated moderate levels of genetic diversity , parameters that decreased significantly when simulations were performed in P. caesium. Additionally, a genetic structure of the populations was found with low gene flow (Nm = 0.92). Genetic parameters are negatively affected by the loss of individuals and the local extinction of populations. Based on our results, we propose to reclassify the conservation status of the species in danger of extinction, hence a series of conservation strategies are provided to prevent its disappearance.Genetic studies in rare species are important to determine their patterns of genetic diversity among populations and implement conservation plans aimed to reduce extinction risks.  Ecological and genetic studies are essential to implement conservation plans, especially in endangered, rare and endemic plant species ,2,3, in Currently, there are various methods of DNA-based markers for the genetic analysis of plant populations, the most common being amplified-fragment length polymorphism (AFLP), inter-simple sequence repeats (ISSRs), microsatellites (SSR), and single nucleotide polymorphisms (SNPs); each varies in polymorphism quantification, repeatability, resolution power and analysis cost ,10,11. AAlthough genetic studies are important toward the species conservation, these results are rarely included in the Red List of threatened species of the International Union for Conservation of Nature’s (IUCN) methodology ,15,16,17Nowadays, habitat loss and climate change are two of the major causes of the current biodiversity crisis, as they cause not only cascading extinctions ,2,3, butPachyphytum, an endemic genus of the Crassulaceae [Pachyphytum caesium Kimnach & Moran is a cliff-dwelling species endemic to the dry tropical forest of Aguascalientes in central Mexico, and has been categorized as PSESP due to its highly specialized habitat, small and fragmented populations, and a high vulnerability to environmental changes [Habitat loss has been particularly worrying in central Mexico, due to land-use change, as more than 80% of its original vegetation has been transformed into agriculture and cattle raising ,24. Thissulaceae . Knowledsulaceae ,27,28,29 changes . Based o changes  classifiThe MER is a tool that is applied exclusively in Mexico, as part of the Mexican Official Standard (NOM-059-SEMARNAT-2010) . This toPachyphytum caesium reported by Clark-Tapia et al. [Because an appropriate estimation of extinction risk is one of the key components of any conservation plan, in this work we evaluated the diversity and genetic structure of the currently know populations of a et al. . In addiP. caesium are located in this region [2, representing 3.18% of the state’s total surface [The study area is located in central Mexico, between 22°40′00″ N and 102°30′00″ W in the southwestern portion of the state of Aguascalientes . The onls region , whose n surface .Pachyphytum caesium (Crassulaceae) is a succulent perennial plant growing in the north-central region of Mexico [f Mexico ,28. A gef Mexico ,28,29 wif Mexico . Populatf Mexico . The areP. caesium can be propagated through asexual reproduction, from the leaves or stems that are detached (Pers. Observations).This species presents two flowering periods per year (from February to April and from September to November). The reproduction is sexual, and the observed pollinators are bees and hummingbirds. The fruit produces numerous small (<1 mm) brown seeds. However, their reproductive biology is unknown, and more studies are needed . AdditioPachyphytum caesium was carried out between 2009 and 2013 as new populations were found. We collected young leaves of 20 individuals per population, for a total of 120 individuals. Distance separation between collected individuals was 20 m to avoid kinship. To access each site and collect the tissue, we used a rock-climbing technique with a Petzl stop descender. Subsequently, we stored the collected biological material in a plastic bag. Once in the laboratory, samples were stored at −70 °C to preserve it until used.The tissue collection of DNA was extracted following the Qiagen DNeasy Plant Kit protocol . For the evaluation of the quality and quantity of genetic material obtained from the 120 individuals, electrophoresis was performed in 1% agarose  gels in TBE buffer 0.5X, and stained with Midori Green Direct  for an average of 40 min at 90 V. Subsequently, these gels were visualized with UV light in a transilluminator equipment Gel Doc XR + System .P. caesium, we opted for ISSR, a dominant marker with reasonable reproducibility and polymorphism than other markers, as well as a robust pattern in the analysis of genetic diversity [P. caesium individuals; these were selected for their high-polymorphic states and repeatability , effective number of alleles per population (Eff_num), and private alleles per population (Ie) were determined as the total number of observed bands (It) using GenoDive v.3.0 . In addiMoreover, we calculated the following genetic diversity parameters: (1) percentage of polymorphic loci with 95% criterion (%P) and (2) expected heterozygosity (He)  using thWe calculated the population structure in six ways: (1) the level of genetic differentiation among populations was obtained with the estimator Gst , using tAdditionally, we used (4) the principal component analysis (PCA) to analyze genetic variation and possible associations in the allele frequencies of the sampled populations using a covariance matrix . (5) We P. caesium. It was performed using the software STRUCTURE v.2.3.4 [Finally, (6) we used a Bayesian model-based clustering approach to identify the genetic structure in populations of  v.2.3.4 . We used v.2.3.4 , which i v.2.3.4  to deterP. caesium shows a pattern of isolation by distance, a Mantel test with 10,000 permutations was performed with the APE package [To evaluate whether  package , using t package . Subsequ package .In 2018, we returned to the previously collected populations and found a drastic decrease in abundance (≥50% in two sites), in addition to an extinct local population reported by Clark-Tapia et al. . Given tPachyphytum caesium. This tool used four normalized criteria to assess risk categories of species: (A) breadth distribution; (B) habitat status; (C) intrinsic biological vulnerability ; and (D) human impact. Numerical values of each criterion are assigned in ascending order of risk, which are subsequently added up. Each criterion is evaluated qualitatively, and the total score is calculated by adding the normalized results from the four. The higher the total score value, the higher the risk of extinction of the assessed species. According to the MER [Based on the method for evaluation of the risk of extinction of species in the Mexican legislation  , we esti the MER , species50) and local extinction (2020eex). In addition, we integrated ecological and genetic data, considering average values (2020e + g) and two simulations, one with loss of individuals (2020e + g50) and the other with extinction (2020e + gex). For comparison, we also applied parts of the IUCN Red List system [We analyzed the MER for two years (2005 and 2020e) based on the ecological and genetic knowledge of the species. We used 2005 because we only had basic knowledge (breadth distribution and habitat status). In 2020e, we analyzed the MER criteria using only ecological knowledge  and ecolt system . This met system .Pachyphytum caesium that presented the highest number of alleles was Presa Cebolletas (96), and the lowest was Presa Malpaso and Mesa Montoro . The total number of alleles did not change when performing the simulation with random extraction of 50% of the individuals or the local extinction of two populations (Presa Cebolletas and Mesa Montoro). However, the number of private alleles decreased significantly   .On the other hand, the highest levels of genetic diversity were found in Presa Cebolletas , and the lowest in Mesa Montoro , with a low population average . The expected heterozygosity was significantly more affected by the simulated extraction of individuals than by the local extinction of two populations . Furtherp < 0.001) partitioning among the populations, of which 43% was attributed among populations and the rest 57% within populations. The coefficients of genetic differentiation among populations Gst and Fst were 0.471 and 0.443, indicating that more than 44% variation was presented among the populations, which is indicative of population structuring. Identical values of variation between populations  were obtained after the random extraction of individuals or the local extinction of populations.The analysis of molecular variance (AMOVA) showed a significant  explained 23.98% of the accumulated variance, where the first component explained 13.56% of the total variation, while the second explained 10.42%. The analysis generated four well-defined population groups, including the association between Puente Cuates, Presa Malpaso, and Río Gil, which is closely associated with Mesa Montoro a.In the PCAs obtained with the simulations of the extraction of individuals b and theFinally, discriminant analysis of principal components (DAPC) detected that the optimal number of principal components to generate the model was two. This result generated a model of two discriminant functions  where a grouping pattern between populations like the PCA was obtained and there was a strong structuring between populations a.With the simulations of individual extraction b, the grP. caesium populations , and mean (similarity score) were detected at K = 2 by Evanno’s method using the total individuals and simulated 50% of populations, indicating that two groups could be distributed across all the ulations a,b. A deulations . Under tulations c.Gene flow between the populations was Nm = 0.92 migrants per generation for all individuals or considering 50% of the individuals per population. However, Nm decreased dramatically to 0.633 when we simulated a local extinction of two of the six populations.p = 0.29; 999 permutations), or 50% of the individuals , and the local extinction of two populations .Mantel’s correlation analysis found no correlation between genetic and geographic distances considering the six populations  (The MER showed four results according to the criteria and knowledge used in the method: (1) with basic knowledge of two criteria (A: breadth distribution and B: habitat status), the species was classified in special protection category (2005). (2) Adding the ecological knowledge about intrinsic biological vulnerability and human impact , the status of the species was elevated to threatened (2020e). (3) After incorporing the genetic knowledge into intrinsic biological vulnerability , the status of the species was further elevated to in danger of extinction level (2020e + g). Although the inclusion of genetic factors in both simulations also generated a status of danger of extinction, the total score in vulnerability was higher (2020e + ge + gex) .50 and 2020e + g50), and (3) the critically endangered status achieved with simulations of local extinction (2020eex and 2020e + gex).On the contrary, the IUCN system showed three conservation statuses: (1) the status of vulnerable for years with basic knowledge (2005), ecological (2020e), and ecological-genetic (2020e + g). (2) The endangered status obtained in the simulations with a decrease in population size ; however, it shows an absence of sexual recruitment despite producing seeds. For this reason, we suggest that the small populations’ size of ulations . The hisulations  and is rulations . Its higulations . Furtherulations ,64. ThisDraba aizoides, a cliff-dwelling species of the Northern Franconian Jura and Swabian Alps in Germany due to the decrease in the population number of adults and seedlings. Our results suggest a similar scenario of affectation at the genetic level, considering the simulation results with loss of individuals and local extinction. It is known that the populations of P. caesium have decreased due to changes in land use and sport climbing [Land-use change or anthropogenic activities have been documented to affect the distribution and size of a population and species’ genetic diversity ,23,54,65climbing ; howeverPachyphytum caesium, like other endangered or endemic species, such as Megacodon stylophorus (Gentianaceae) [Rhodiola alsia (Crassulaceae) [P. caesium populations have a fragmented distribution due to land-use changes, with a discontinuous distribution associated with cliffs in the remnants of the tropical dry forest in the state of Aguascalientes [Pachyphytum makes its comparison difficult. However, it is possible with other endemic plants or PSESP that inhabit cliffs, including members of the Crassulaceae family. In these species, it is common to find a high genetic differentiation between populations, e.g., [An incipient level of among-population genetic differentiation was revealed in anaceae)  and Rhodulaceae) . The genulaceae) ,54. P. calientes . The lacs, e.g., ,54,61,66Pachyphytum caesium, the absence, or low gene flow, as a factor that could homogeneize allelic frequencies [P. caesium populations, added to the allelic loss due to the decline in their populations and local extinctions, increases the genetic drift effect and allows the action of other evolutionary forces as inbreeding. If this is so, new molecular studies (SNPs) are also required to know aspects of inbreeding, genetic landscape, and locate local and functional genotypes to provide greater certainty in the conservation of the species.In quencies ,67, togequencies ,69. GeneEven though there is a discrepancy in the conformation of groups between the Minimum spanning network, PCA and DAPC analyses showed between 3 and 4 groups, concerning STRUCTURE that obtained two groups, or three in the simulation without two populations. At landscape level, the cliffs from Río Gil, Presa Malpaso, and Puente Cuates have greater connectivity between them as they are within the same micro-basin and are more likely to exchange migrants. Therefore, they have a greater association and overlap extensively in the analyses. In contrast, the three remaining populations have been isolated in recent decades with less probability of genetic exchange.P. caesium populations were fragmented, leading to isolation and genetic drift phenomena as reported in other endemic species [Cluster analysis and STRUCTURE clustering showed a clear geographic pattern suggesting that, in the past,  species . A possi species , furtherP. caesium, a result found in other studies, which also emphasizes the need to consider genetic studies.This study highlights the importance of using genetic data in the risk analysis of a species, as suggested by Garner et al. , VictoriLike the MER, the risk is increased in the IUCN by modifying the population size and distribution area; two key criteria used by these methods assign threat status ,30. AlthP. caesium, precautionary conservation measures should be appropriate because the risk of extinction is higher for small populations [In PSESP, as in ulations ,23, whiculations  and the ulations . That isP. caesium is not subject to federal protection in the short term, its vulnerability to adapt to future environmental changes increases, mainly due to a decrease in private alleles, e.g., [The immediate conservation strategy is to insert the species on the IUCN red list and the list of species at national risk, in addition to the studies of the species’ vegetative propagation and reproductive biology which have already begun. The establishment of rappelling routes and the prohibition of this activity in some areas where the species is found is a pending activity. Given the possibility that s, e.g., , and reds, e.g., , which wPachyphytum caesium exhibit moderate to low levels of genetic diversity, a strong genetic structure of their populations, and low gene flow. Genetic drift is an evolutionary force that negatively affects the genetic population structure and depends on the demographic stability of the species. The simulations of extraction of individuals and the local extinction of populations negatively affect genetic diversity, which should be considered for population conservation. According to ecological and genetic data, populations of P. caesium are at risk of extinction following the MER. We highlight the importance of using genetic data in MER, which is why we suggest its use in the IUCN Red List methodology. Finally, genetic data can support future endangered species conservation planning.Our study found that


The sample of this study was obtained through a random sampling technique to determine the treatment (n = 32) and control (n = 34) groups. Before and after implementing the TBT program, glial cell-derived neurotrophic factor (GDNF) level measurement and cognitive function and postural balance assessment were performed. Cognitive function was measured by using Montreal cognitive assessment (MoCA). Postural balance was measured in two ways by using the timed up and go (TUG) test and Tinetti performance-oriented mobility assessment (POMA). The treatment group showed significantly greater changes than the control group in GDNF levels (2.24 (±0.63) vs. 1.24 (±0.43), P = 0.001), cognitive function (24.66 (±3.42) vs. 19.18 (±2.67), P = 0.001), and postural balance (TUG [14.00 (±4.04) vs. 18.68 (±3.98)]; POMA [26.53 (±1.74) vs. 23.47 (±3.06)], P = 0.001) after training. The treatment group also showed a significant relationship between GDNF levels and cognitive function  and postural balance . The TBT program affects the levels of GDNF in old people. The TBT program involves cognitive function improvement and affects postural balance changes in old people.Exercise in the form of physical activity can provide neuroprotective benefits. The purpose of this study is to determine the effect of the task balance training program (TBT program) on the glial cell-derived neurotrophic factor levels, cognitive function, and postural balance in old people. The population of this study was the old people members of the Batara Hati Mulia Gowa Foundation who were willing to participate in the study ( Aging is a gradual process of the declining ability of tissues to repair or regrow themselves and maintain their normal functions. As the world's population rises, the population of older people is expected to continue to rise as well. The aging process causes continuous changes in old people's bodies including a gradual loss of function, weakness, illness, and even mortality. In old people, there is a decrease in muscle mass by 0.5%-1% per year, resulting in reduced strength and declining physical condition. Inactivity and the lack of exercise may exacerbate the physical condition of old people [There are several types of degenerative diseases affecting the old people population, such as brain nerve damage or cognitive decline. A person is categorized as having a declining cognitive function, commonly known as dementia or senility, if s/he shows 3 or more of the following symptoms without disruption of consciousness: having attention and memory disturbances, place and time disorientation, and construction and execution inabilities (such as making decisions and solving problems). The cognitive function typically deteriorates as we age. As a result, this decline will impact the daily life of old people , 3.Impairments in mobility and cognition are common in many neurological conditions, making each movement require more attention. Physiologically, attention is divided into two types: selective attention  and divided attention . Divided attention is more often affected by neurological conditions. However, divided attention is necessary to perform two tasks simultaneously, such as walking and speaking , 5.In everyday life, the ability to do work at the same time is necessary and expected. This represents the superiority of human evolution. However, when processing several stimuli and producing different responses, often there is a lower performance in one of the tasks being performed. Exercises are designed to improve walking and daily function in old people . More and more rehabilitation strategies employ motor training, which facilitates patients by integrating functional activities to improve daily abilities. The task balance training (TBT) program is an advanced development of the strength and balance techniques and exercises with systematic procedures of repetitive muscle contractions. TBT can be classified into Single Task (ST) and Dual Task (DT). Some studies have proven that DT training was effective to improve balance and gait , 2. It cMuscle adaptation in the loading process causes muscle hypertrophy as the result of the training. Exercise can also improve joint flexibility and range of motion and stimulate proprioception by increasing motor unit recruitment, which activates the Golgi tendon organs and muscle spindles. The more the number of muscle fibers innervated by a motor nerve, the greater the strength of the muscle. Throughout the training, the intrafusal and extrafusal threads receive sensory information, which will be relayed and processed by the brain to determine the necessary amount of force of the muscle cocontraction. Some of the responses will return to the extrafusal fibers and activate the Golgi tendon, resulting in better coordination between intrafusal and extrafusal fibers and afferent neurons in the muscle spindles and thus better proprioception. Sensory input, proprioception, nervous system, and muscular strength contribute to old people's balance –8.Exercise is a physical activity that can provide neuroprotective benefits of increased neurogenesis, synaptogenesis, angiogenesis, and modulation of neurochemical levels in the brain. This process is mediated by a neurotrophic factor, glial cell-derived neurotrophic factor (GDNF), which is involved in neural plasticity, particularly in brain trauma and cognitive and memory impairment. Elevated GDNF levels can increase nerve cell survival in the nigrostriatal and other areas of the cerebral cortex. Activation of dopaminergic neurons produces neurotransmitters on the pathway to the basal ganglia to control body movements. Neurotransmitters are distributed in the prefrontal cortex which functions as memory storage and in the frontal lobe to maintain cognitive functions such as problem-solving, higher-order thinking, and learning –11.β (TGFb) superfamily, along with three other structurally similar factors—neurturin, artemin, and persephin. GDNF is also involved in the hippocampal synaptogenesis, playing a role as an instructional factor by activating presynaptic locations. The GDNF family receptor-1 (GFRa1) complex is needed for proper hippocampal circuit formation [GDNF is currently recognized as a critical component of nigrostriatal neuron development, maintenance, and protection and as a possible factor in the maintenance and repair of dopaminergic neurons damaged by Parkinson's disease. GDNF is a member of the neurotrophic factor family, a subfamily of the transforming growth factor-ormation , 12. TheThe population in this study was the old people members of the Batara Hati Mulia Gowa Foundation who were willing to participate in the study. Inclusion criteria are (a) those who have not had difficulty walking or fall within the last 3 months, (b) those who have no serious pain, musculoskeletal damage, or neurological damage, (c) those who understand the instructions of the examiner and can perform them, and they were between the ages of 60 and 75. Exclusion criteria are (a) a person with acute inflammation and (b) a person with visual and hearing problems. All participants understood the content of the study and participated voluntarily. The criteria for considering types of people included in studies should be sufficient to encompass the likely diversity and ensure the sample can carry out exercise without restriction of movement ability. The elderly have difficulty performing exercise movements such as walking if they have serious pain, muscle, and nerve disorders. The elderly with walking disorders, a history of falls, and muscle and nerve disorders can affect balance when they are given exercise; it can be limited to carrying out exercise movements such as walking exercises and standing exercises on one leg in a series of exercises. The random sampling approach was utilized to determine the participants of the treatment and control groups. The treatment group was assigned to the TBT program, while the control group was provided with therapeutic communication. It is an effective communication that is carried out consciously and designed in such a way by medical personnel to patients with the aim of helping patients to get positive health outcomes as expected [Blood sampling collection, Montreal cognitive assessment (MoCA), timed up and go (TUG) test, and Tinetti performance-oriented mobility assessment (POMA) were applied in both groups before and after intervention, respectively. This study received approval from the Health Research Ethics Commission, Faculty of Medicine, Hasanuddin University (828/UN4.6.5.31/PP36/2020). The research ethics recommendations included a requirement to provide an informed consent form and agreement sheet to participants who met the inclusion criteria. If the subject were willing to become a participant, they must sign the consent form. The sample who refused to participate would not be forced, and their rights would be respected. The second recommendation was anonymity, which required the author to give a specific code to each participant instead of listing participants' real names. Thirdly, the author must ensure confidentiality of the participants' information and only reported relevant information in the research results.The exercise program was performed 3 in three sessions a week for 4 weeks with 30 minutes per session. The exercise program comprised cognitive exercises and motoric exercises. Each exercise was performed for 30 sec followed by 30 sec of rest, and three sets were performed with 1 min break between the sets. The exercise intensity was 30-40% of the elderly's maximum heart rate range. The intensity complies with the American College of Sports Medicine guidelines for exercise testing and prescription when it comes to exercise recommendations for old people. This approach does have beneficial effects related to health, especially in the elderly. The details of the exercise program are shown in μL of the serum was stored in a sample tube. The GDNF levels were assessed using the enzyme-linked immunosorbent assay (ELISA) method with the GDNF ELISA kit , respectively. Cognitive function was measured by using Montreal cognitive assessment (MoCA). Postural balance was measured in two ways by using the timed up and go (TUG) test and Tinetti performance-oriented mobility assessment (POMA). The timed up and go (TUG) test was conducted to evaluate changes in dynamic balance following the intervention. The subject began in a seated position. At a start signal, they stood up, walked 3 m away, turned around, walked back, and returned to a seated position. The time was measured from the start signal to when the subject was seated in the chair again.The instruments used to measure the variables studied consisted of a Riester mercury sphygmomanometer, OneMed stethoscope, questionnaire, informed consent, and stopwatch. The instruments for blood sampling collection consisted of a 3 cc syringe, 25 G wing needle, tourniquet, 3 cc EDTA tube, plaster, and alcohol cotton ball. The instruments for storing blood samples consisted of a centrifuge, Eppendorf tube, filter tip, micropipette, and -20°C refrigerators. To measure GDNF levels, the enzyme-linked immunosorbent assay (ELISA) test was performed using a GDNF kit. Blood tests were performed on all subjects after 12 h of fasting. The blood was drawn at the same time under the same conditions before and after the intervention. A disposable needle was used to draw 3 mL from the anterior cubital vein. The blood was stored in serum-separating tubes for serum and centrifuged for 10 min at 3,000 rpm; 200 t-test was also conducted to determine the effect of the TBT program on GDNF levels, cognitive function, and postural balance. Paired sample t-test measured the difference experienced by the subjects from the same group after being subjected to treatment. Furthermore, to find the mean difference of each variable, an independent sample t-test was performed. The independent sample t-test was carried out to examine if there was a statistically significant mean difference between the two independent groups. To examine the correlation between GDNF levels with cognitive function and postural balance, Spearman's correlation test was performed using the SPSS program. The Spearman's correlation test was selected in this study because the type of data to be processed from both independent and dependent variables was ordinal data. Spearman's correlation test was also able to provide the correlation coefficient value and show the direction of the relationship, whether it was negative or positive.The data were analyzed using univariate analysis (frequency distribution) to produce the distribution and percentage of each variable. Paired The participant' characteristics are presented in P > 0.05; thus, GDNF levels in the elderly obtained were normally distributed; cognitive function data distribution in the elderly obtained P > 0.05; thus, cognitive function in the elderly obtained was normally distributed, and balance data distribution in the elderly obtained P > 0.05; thus, the balance in the elderly obtained was normally distributed. A homogeneity test was carried out whose results showed that the significant value of Levene's test for equality of variances was more than 0.05, so it can be concluded that the data values of GDNF, MoCA, TUG, and POMA of the intervention group and the control group were homogeneous.Normality distribution tested with Shapiro-Wilk test on GDNF levels in the elderly obtained P value < 0.001, indicating that the TBT program influenced the GDNF levels. Cognitive function was significantly improved in the treatment group with a P value < 0.001, indicating that the TBT program affected cognitive performance. With a P value < 0.001 for both the TUG and POMA scores, it can be concluded that there is a significant improvement in postural balance in the treatment group, suggesting that the TBT affected postural balance (The mean GDNF level before treatment is 0.90 (±0.31). The GDNF level rises to 2.24 (±0.63) after treatment, suggesting a statistically significant difference in the mean GDNF levels before and after treatment . The mea balance .P < 0.001)  and 1.24 (±0.43), respectively. The mean values of cognitive function for the treatment and control groups are 24.66 (±3.42) and 19.18 (±2.67), respectively. The mean TUG scores for the treatment and control groups are 14.00 (±4.04) and 18.68 (±3.98), respectively. The mean POMA scores for the treatment and control groups are 26.53 (±1.74) and 23.47 (±3.06), respectively. Therefore, it can be concluded that there is a statistically significant difference in the mean POMA score between the treatment and control groups. All variables measured in this study showed a statistically significant difference between the treatment and control groups (< 0.001) . In otheComparison of the GDNF, MoCA score, TUG score, and POMA level score in pretest and posttest in the two groups revealed that the levels of the treatment group were not considerably different from those in the control group in the pretest. In the posttest, the levels of GDNF, MoCA score, and POMA score in the treatment group were considerably higher than those in the control group, and TUG score was lower in the treatment group, as shown in P < 0.001 signifies a relationship between GDNF levels and cognitive function. The correlation coefficient values of 0.818 (TUG) and 0.630 (POMA) indicate a correlation between GDNF levels and postural balance. According to the degree of closeness between the two variables, a strong relationship and positive linear pattern indicate that the higher one's GDNF levels, the better one's postural balance will be. The value of P < 0.001 signifies a relationship between GDNF levels and postural balance, either measured using the TUG test or POMA . This indicates that one or both abilities have weakened due to declining metabolism in the frontal area. It is necessary to combine physical activity with cognitive stimulation; hence, a TBT program was designed to enhance cognitive capacities and slow brain aging by engaging cognitive and motor stimulations. Physical activity can provide neuroprotective benefits mediated by GDNF. GDNF is an essential protein that is required to develop, maintain, and protect nigrostriatal neurons, particularly in protecting and restoring dopamine neurons affected by Parkinson's. Therefore, it is necessary to determine whether the TBT program affects GDNF levels, cognitive function, and postural balance in old people. The effect of the TBT program on the GDNF levels, cognitive function, and postural balance in old people will be discussed according to the research objectives, as follows.Glial cell line-derived neurotrophic factor (GDNF) supports neuroplasticity in the neuromuscular system from childhood to maturity. To maintain a healthy neuromuscular system, a steady supply of neurotrophic factors (NF) is required to support motor neuron growth and maturation. NFs contribute to the survival of motor neurons throughout their lives by encouraging their growth during embryonic development, maintaining them till adulthood, and regenerating them after injury . In adulP < 0.05). GDNF is activity-dependent, and by changing the type and intensity of exercise, it is possible to increase the protein content of GDNF in slow and fast-twitch muscle fibers. This also strengthens the notion that GDNF affects the NMJ structure in slow and fast-twitch muscle fibers. Exercise may enhance the protein content of GDNF in skeletal muscle, resulting in improved NMJ plasticity and neuromuscular health [P < 0.05). These findings suggest a rise in the GDNF protein content in the muscles of the treated mice. It can be inferred that intensified physical activity increases structural neuroplasticity in the neuromuscular junction (NMJ) elements, resulting in improved neuromuscular function [The results of this study are in line with the study conducted by Gyorkos et al.  on rats r health . Likewisr health , long-tefunction .The training protocol was adequate to produce an increase in GDNF level, providing additional evidence for activity-dependent neurotrophic support mechanisms. The CSA of SOL muscle fibers reduced in response to this exercise plan shows that these muscles are engaged during this activity. The reduction in fiber diameter caused by training is an indication of recruitment. It can be viewed as an excellent adaptation to a faster oxygen, carbon dioxide, and waste product exchange, which prolongs fatigue time , 14.A high level of cognitive function is critical for integrating and interpreting sensory-motor information necessary for maintaining equilibrium in the everyday environment. Individuals frequently complain about their memory as they age. However, compared to memory, other cognitive domains such as processing speed, executive function, and attention are more affected, which can have an impact on sensorimotor functions. Executive function and attention deficits have been linked to deteriorating physical function, such as slower gait and poor balance  , and 5-hydroxytryptamine (5-HT) [The findings of this study are supported by the survey conducted by Jardim et al. , showinge (5-HT) .Endogenous GDNF is necessary for cognitive abilities, as shown in a study of heterozygous mice with targeted GDNF gene deletion which resulted in a reduced spatial learning capacity . In the P < 0.001) [There is mounting evidence that GDNF levels may play a role in the etiology and progression of illness, as they do in Alzheimer's disease with cognitive impairment. GDNF levels were shown to be strongly associated with cognitive impairment (< 0.001) . The stu< 0.001) , who att< 0.001) . GDNF pr< 0.001) . GDNF in< 0.001) , 18. GDN< 0.001) . The inc< 0.001) , 21.Walking and falling disorders are problems that often occur in old people patients. In old people, there is a physiological decline in the musculoskeletal system, which manifests in a reduction of the number and size of muscle fibers, resulting in a weaker lower extremity muscle, endurance, coordination, and limited range of motion (ROM). Balance is a complex positional defense against outside interference. Impaired balance and gait and weakness of lower extremity muscles can cause falls in old people , 23.Falls are the main cause of morbidity in old people patients. The TUG test is a sensitive and objective method of assessing balance and walking disorders. TUG test evaluated the time taken to complete the entire series of tests. Tinetti POMA, on the other hand, can provide additional benefit as the assessment minimize falls and at the same time train physical balance in old people. Physical balance exercise is an exercise to control movement and position in the center of the body. This exercise is also an essential component in maximizing other physical activities .Physical activity appears to help prevent premature death of dopamine cells. In the basal ganglia, exercise-induced improvement in the synthesis of GDNF has been observed. In parkinsonism mice (or) monkeys, motor deterioration can be delayed and dopamine levels in the brain can be preserved by running on a treadmill , 24.The results of this study are in line with the study conducted by Marques et al. , which sIn conclusion, the task balance training program improved GDNF, cognitive function, and postural balance in old people. Therefore, it could be suggested as an intervention program for improving the cognitive function and postural balance in old people. The limitations of this study were a relatively small sample size, the intervention period was short, and no laboratory examination of neurotrophic factor receptors which specifically bind to GDNF that might affect. Further study should be conducted to analyze the role of GDNF receptors through exercise on cognitive function and postural balance in old people.