Aedes aegypti–a primary vector of dengue, Zika, and chikungunya viruses. We then validated the use of our approach to generate experimental microcosms colonized by standardized lab- and field-derived bacterial communities. Our results overall reveal minimal effects of cryopreservation on the recovery of both lab- and field-derived bacteria when directly compared with isolation from non-cryopreserved fresh material. Our results also reveal improved reproducibility of bacterial communities in replicate microcosms generated using cryopreserved stocks over fresh material. Communities in replicate microcosms further captured the majority of total bacterial diversity present in both lab- and field-based larval environments, although the relative richness of recovered taxa as compared to non-recovered taxa was substantially lower in microcosms containing field-derived bacteria. Altogether, these results provide a critical next step toward the standardization of mosquito studies to include larval rearing environments colonized by defined microbial communities. They also lay the foundation for long-term studies of mosquito-microbe interactions and the identification and manipulation of taxa with potential to reduce mosquito vectorial capacity.Mosquitoes develop in a wide range of aquatic habitats containing highly diverse and variable bacterial communities that shape both larval and adult traits, including the capacity of adult females of some mosquito species to transmit disease-causing organisms to humans. However, while most mosquito studies control for host genotype and environmental conditions, the impact of microbiota variation on phenotypic outcomes of mosquitoes is often unaccounted for. The inability to conduct reproducible intra- and inter-laboratory studies of mosquito-microbiota interactions has also greatly limited our ability to identify microbial targets for mosquito-borne disease control. Here, we developed an approach to isolate and cryopreserve bacterial communities derived from lab and field-based larval rearing environments of the yellow fever mosquito  i) facilitate the study of mosquito traits of interest  in the absence of confounding effects of microbiota variation, and (ii) enable reproducible intra- and inter-laboratory studies of mosquito-microbiota interactions to identify microbial targets for disease control. Our results are also of broad interest to researchers in the fields of microbial ecology and host-microbe interactions because they demonstrate how tools commonly used to study microbiota assembly and function in mammals can be leveraged in other systems.Mosquitoes develop in the presence of diverse bacterial communities that shape their ability to transmit disease-causing pathogens. However, our current understanding of mosquito-microbiota interactions is largely based on studies of individuals colonized by low-diversity communities of bacteria that are not commonly associated with mosquitoes in the laboratory or field. In this study, we developed an approach to isolate and cryopreserve microbiota from mosquito larval rearing environments in the lab and field. We then demonstrated the successful use of this approach to produce experimental microcosms colonized by standardized microbial communities. Our results are of critical significance to the field of vector biology because they will directly ( The community of bacteria (hereafter referred to as ‘microbiota’) present in the aquatic environments where mosquito larvae develop can have profound impacts on mosquito biology by modulating larval growth and development and, consequently, adult survival, reproduction, and the competency of adult female mosquitoes to transmit human pathogens (reviewed in ). HoweveAedes aegypti and Culex quinquefasciatus mosquitoes that serve as primary vectors of arboviral infections and filariasis in humans   .We initially sought to develop an approach to isolate and cryopreserve microbiota from conventional larval rearing pans in the laboratory for recapitulation in experimental microcosms. Multiplex sequencing of 16S rRNA gene amplicons for the resulting rearing pan- and microcosm-derived water samples generated a total of 2,669,979 quality-filtered reads with a median sequencing depth of 44,707 reads per sample . An unusi.e., those with a maximum relative abundance ≤1% in rearing pans), with significantly fewer rare ASVs being recovered in experimental microcosms generated using cryopreserved stocks of lower cell densities , even at 5 days post-incubation  between any of the microcosms we generated  of reads from these samples were assigned to ASVs belonging to one of eight bacterial families within the following phyla/classes: Alphaproteobacteria , Betaproteobacteria (Comamonadaceae), Gammaproteobacteria , and Bacteroidetes  . A totalcubation . Howeverenerated .4, 105)  . Differences in beta diversity, measured as average Bray-Curtis dissimilarity, were also overall higher between rearing pans and unprocessed controls than between rearing pans and experimental microcosms generated using material from cryopreserved stocks, regardless of cell density  . Further= 0.001) , post-ho density .i.e., decreased in abundance) in response to cryopreservation .Finally, we performed ALDEx2 tests to identify rearing pan ASVs that significantly changed in relative abundance in response to cryopreservation and/or re-culturing under conventional mosquito rearing conditions Figs  and S4. ion Figs  and S4. Next we sought to validate the use of cryopreservation approaches developed above to generate experimental microcosms colonized by microbiota derived from a naturally occurring mosquito larval habitat in the field. Multiplex sequencing of 16S rRNA gene amplicons for the resulting habitat- and microcosm-derived water samples generated a total of 2,111,045 quality-filtered reads with a median sequencing depth of 43,541 reads per sample , with raP = 0.426).As expected, we identified a substantially higher diversity of bacteria (993 ASVs) across the field-derived water samples as compared to those derived from conventional larval rearing pans in the laboratory, with ~42% of reads being assigned to ASVs belonging to the eight bacterial families dominating communities in rearing pans and the remaining ~58% being assigned to ASVs belonging to one of 306 other families within 187 bacterial orders and 34 phyla/classes . A substP > 0.05).ALDEx2 tests identified a total of 110 of the 993 ASVs found in the habitat water samples we sequenced showing significantly different relative abundances in experimental microcosms, but only 12 of these ASVs, representing <6% of all habitat water sequences, were specifically negatively affected in response to cryopreservation Figs  and S7. While our isolation and cryopreservation approaches resulted in the loss of some lab- and field-derived bacterial taxa and concurrent changes in the total alpha and beta diversity of microbiota within experimental microcosms as compared to the larval rearing pans and habitat water we originally sampled, we observed little variation among the bacterial communities present within replicate microcosms generated using the same cryopreserved stock, with the only exceptions being those generated using stocks of lower cell densities . On averMosquitoes live in close association with bacteria and other microorganisms that shape their ability to transmit pathogens . Howeveri) decouple the effects of sample processing and the cell density of cryopreserved stocks on the recovery and persistence of bacteria in microcosms from those of larvae, which are known to shape water pH, nutrient levels, and even bacterial metabolism [ii) better reflect natural microbiota exposure and acquisition processes in the field, whereby larvae hatch from eggs laid directly into (or adjacent to) water containing established bacterial communities [Ae. aegypti revealed a bacterial community comprised of ~100 unique bacterial taxa and dominated by members of the Proteobacteria, Bacteroidetes, and Firmicutes, consistent with sequencing studies of other laboratory colonies of mosquitoes [Paenarthrobacter, which were significantly reduced or absent in experimental microcosms and may represent taxa that are not readily amenable to centrifugation and/or cryopreservation using our methodology.In this study, we first developed an approach to isolate and cryopreserve microbiota from conventional larval rearing pans in the laboratory for recapitulation in experimental microcosms. We intentionally conducted our experiments in the absence of any larvae in order to:  detected in laboratory rearing pans and commonly detected in field-collected mosquitoes –49,60–65The cell density of cryopreserved stocks also had a much larger impact on the recovery and persistence of field-derived taxa, with microcosms generated using stocks of lower cell densities exhibiting more variable bacterial communities that were overall lower in complexity that communities in microcosms generated using stocks of higher cell densities. This is consistent with results showing that small sampling volumes from aquatic and other high-complexity environments like soil often fail to adequately capture bacterial diversity –77, owine.g., immediately after a rainfall). Defined diets that support mosquito growth and development could also be developed to improve the reproducibility of field bacterial diversity between replicate microcosms, including the maintenance of rare taxa important for community assembly and stability [Overall, experimental microcosms generated using cryopreserved stocks containing lab-derived microbiota exhibited levels of reproducibility that were the same or better than those present under conventional rearing conditions in the laboratory, highlighting the value of our approach for intra-and inter-lab studies of mosquitoes in the presence of standardized microbial communities. Similar levels of reproducibility could also be replicated in microcosms generated using cryopreserved stocks of field-derived microbiota, although reproducibility significantly decreased with decreasing inoculum size–consistent with our previous observation that variability among the bacterial communities present within replicate microcosms was significantly higher in those generated using cryopreserved stocks of lower cell densities. Future studies are warranted to establish best practices for field habitat sampling, though our results at minimum suggest that higher-volume samples are preferable, and that sampling should be avoided in situations where bacterial density is expected to be low . Unmodified bacteria that naturally inhibit pathogen colonization or mosquito fitness could also be disseminated to mosquito populations. The identification of suitable microbial candidates for pathogen or mosquito control will require a comprehensive understanding of the factors that influence the acquisition, maintenance, and transmission of mosquito microbiota and the mechanisms that underlie how individual microbial species and assemblages impact mosquito vectorial capacity. However, the dearth of tools to manipulate mosquito microbiota, including microbiota derived from naturally occurring mosquito habitats in the field, has greatly slowed progress in this area. In this way, the results reported here not only provide a critical first step toward the standardization of microbial inputs in mosquito studies, but also provide a critical first step toward the identification of taxonomic and functional profiles of bacteria associated with phenotypic traits of interest in mosquitoes. For example, a defined microbiota could be universally adopted by the community to conduct vector competence assays in the absence of confounding effects due to microbiota variation. Libraries of cryopreserved microbiota could also be screened to identify bacteria that improve or reduce mosquito fitness and/or pathogen susceptibility, or to predict the success of individual microbial candidates under variable microbial conditions. Similarly, the results reported here strongly support that our methods could immediately be leveraged to expand studies of mosquito-microbiota interactions to include microbial genotypes derived from the field, and thereby conduct lab-based mosquito experiments with field-relevant microbiota.Owing to their ability to impact numerous components of mosquito vectorial capacity, there is a growing interest in exploiting microbes for mosquito-borne disease control. For example, bacteria that naturally colonize the mosquito gut could be genetically modified to produce effector molecules that alter the mosquito’s ability to become infected with and transmit pathogens, or that reduce mosquito fecundity or lifespan (S1 TableAsterisks (*) indicate samples that were removed from the dataset prior to downstream analyses.(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S1 FigReads from each water library were sampled starting at 1 sequence per step and increased in increments of 100 until the total number of reads per sample was reached. Lines are colored by sample source . Time of sampling of experimental microcosms is designated by line type .(TIF)Click here for additional data file.S2 FigP < 0.05).Proportion of rare (left) and abundant (right) ASVs found in at least one larval rearing pan that were detected in experimental microcosms containing unprocessed water or sterile water plus material from a given cryopreserved stock. An ASV was considered “rare” if it had a maximum relative abundance ≤1% across the four larval rearing pans we sampled, while ASVs with a minimum relative abundance >1% were considered “abundant”. Asterisks (*) indicate significant differences between experimental microcosms generated using cryopreserved stocks relative to unprocessed controls as determined by paired Fisher’s exact tests with Bonferroni correction ((TIF)Click here for additional data file.S3 FigEach ASV is presented with its lowest annotated taxonomic rank (to genus level) together with its ASV ID. The ASVs are color-coded according to the phyla they belong to and plotted according to their effect size, calculated as the levels in samples from experimental microcosms relative to levels in samples from larval rearing pans. Dot sizes correspond to the median centered log-ratio (clr) abundance value for each ASV across rearing pan samples. Dots with a bold red outline represent ASVs that were differentially abundant in both experimental microcosms containing unprocessed water and experimental microcosms containing sterile water plus material from cryopreserved stocks see .(TIF)Click here for additional data file.S4 FigEach ASV is presented with its lowest annotated taxonomic rank (to genus level) together with its ASV ID. The ASVs are color-coded according to the phyla they belong to and plotted according to their effect size, calculated as the levels in samples from experimental microcosms relative to levels in samples from larval rearing pans. Dot sizes correspond to the median centered log-ratio (clr) abundance value for each ASV across rearing pan samples. Dots with a bold red outline represent ASVs that were differentially abundant in both experimental microcosms containing sterile water plus material from cryopreserved stocks and experimental microcosms containing unprocessed water see .(TIF)Click here for additional data file.S5 FigReads from each water library were sampled starting at 1 sequence per step and increased in increments of 100 until the total number of reads per sample was reached. Lines are colored by sample source . Time of sampling of experimental microcosms is designated by line type .(TIF)Click here for additional data file.S6 FigEach ASV is presented with its lowest annotated taxonomic rank (to genus level) together with its ASV ID. The ASVs are color-coded according to the phyla they belong to and plotted according to their effect size, calculated as the levels in samples from experimental microcosms relative to levels in habitat water samples. Dot sizes correspond to the median centered log-ratio (clr) abundance value for each ASV across habitat water samples. Dots with a bold red outline represent ASVs that were differentially abundant in both experimental microcosms containing unprocessed habitat water and experimental microcosms containing sterile water plus material from cryopreserved stocks see .(TIF)Click here for additional data file.S7 FigEach ASV is presented with its lowest annotated taxonomic rank (to genus level) together with its ASV ID. The ASVs are color-coded according to the phyla they belong to and plotted according to their effect size, calculated as the levels in samples from experimental microcosms relative to levels in habitat water samples. Dot sizes correspond to the median centered log-ratio (clr) abundance value for each ASV across habitat water samples. Dots with a bold red outline represent ASVs that were differentially abundant in both experimental microcosms containing sterile water plus material from cryopreserved stocks and experimental microcosms containing unprocessed water see .(TIF)Click here for additional data file.


Simulation is a key component of surgical training, enabling trainees to develop their skills in a safe environment. With simulators broadly grouped into physical models and virtual-reality (VR) simulators, it is important to evaluate the comparative effectiveness of the simulator types in terms of validity as well as cost. The review aims to compare the benefits and drawbacks of novel VR and physical simulators within the broader themes of endourology, laparoscopic and robotic operations, and other urological procedures.Key benefits of bench models include their comparatively lower cost, easy access and provision of haptic feedback, whereas VR simulators are generally self-sufficient, reusable and enable skills of haemostasis to be practised. The advent of perfused 3D printed simulators across a range of urological procedures may replace cadavers as the traditional gold-standard simulation modality.Although possessing differing strengths and downsides, VR and physical simulators when used together can have an additive effect due to skill transferability across the platforms. Further comparative studies are required to directly quantify the differences between physical models and VR simulators in terms of performance metrics and cost-effectiveness. There is lack of validated VR simulators for open and reconstructive procedures. Simulation is an integral part of surgical training enabling trainees to hone their technical skills away from actual patients until they achieve a safe level of competency . This isEndourological procedures include ureteroscopy, cystoscopy and transurethral resections of the bladder and prostate. The SIMULATE international prospective trial  examined3D model simulators have been recently developed in the field of cystoscopy such as the ‘FlexBlad’  and ‘Bla▪▪] examining simulation training in transurethral resection and laser vaporisation procedures of the prostate. The review findings pointed towards VR simulators uniquely possessing data capture and performance evaluation features but that alongside their typically poor haptic feedback, their simulation of bleeding is generally poor for TURP and laser vaporisation. Physical simulators such as bench-top tissue and food-based models also inadequately simulate bleeding for these procedures and carry hygiene risks, but are lower cost and beneficial in that they use real instruments. In the context of laser-based procedures, cadavers demonstrate content validity for holmium laser AEEP training [The European Section of Uro-Technology (ESUT) undertook a systematic review ▪▪ examintraining  with suptraining , VR simutraining ▪,16 and PCNL is another endourological procedure for which simulators have been validated. One study used participant ratings and performance data to compare three simulators ; a porciAlthough cadavers are considered the gold-standard simulation modality, artificial perfusion of cadavers is limited by high cost, positive viral profiles and difficulty clearing the vascular tree. One low-cost, nonbiohazardous physical model for robot-assisted partial nephrectomy (RAPN)  directlyLive animal models such as porcine models pose ethical challenges not associated with other simulators. One study  aimed toVR simulation performance for robot-assisted radical prostatectomy (RARP) has crucially been shown to correlate with live surgical performance in the real operative environment thereby increasing its validity as a training modality. One study  of 20 suWith VR, simulation is moving beyond just basic isolated skills training to full procedural simulation. This has been demonstrated for RARP  with supFor laparoscopic pyeloplasty, various physical simulators exist with advantages over VR simulators such as significantly lower cost and easy access whilst maintaining anatomical realism. As with aforementioned procedures, 3D printed models for pyeloplasty have been produced  demonstrFor the management of urological emergencies such as priapism and testicular torsion, there is an absence of validated VR simulators to develop the skills necessary for the therapeutic interventions these conditions require. There is therefore a reliance on physical simulators to facilitate skill acquisition and these have been incorporated effectively into training curricula. For example, a urological emergency simulation curriculum was devised for military surgeons  utilisinFor penile fracture repair, the first validated simulation model was recently created , based oReconstructive urology is another field in which validated VR training is absent. Instead high-fidelity, 3D-printed penile models have been produced for the simulation of plaque incision and graft (PIG) as indicated for the treatment of Peyronie's disease , achieviTo conclude, physical models are generally superior with regard to cost-effectiveness, the provision of haptic feedback and ease of access, while VR simulators are self-sufficient, reusable and avoid the ethical and infection concerns associated with animal models. Although cadavers have long been considered the gold-standard simulation modality, these may soon be superseded by the advent of highly realistic, perfused 3D printed models capable of procedural simulation incorporating anatomical variation and intraoperative complications  at a subThe authors received no source of funding and have no conflicts of interest.None.Baldev Chahal, Abdullatif Aydin and Kamran Ahmed have no conflicts of interest or financial ties to disclose.


Watermelon stomach, also known as gastric antral vascular ectasia (GAVE), is a rarely seen cause of acute upper gastrointestinal bleeding or chronic iron deficient anemia, revealed when gastroscopy is performed. GAVE affects predominantly women of advanced aged and is usually associated with certain underlying co-morbidities including liver cirrhosis, autoimmune diseases and chronic kidney disease. Based on the above, we report the case of a 75-year-old woman under hemodialysis for six consecutive years due to end-stage renal disease attributable to hypertensive nephrosclerosis. The patient also suffered from chronic ischemic heart disease, severe peripheral artery disease and an unidentified connective tissue-disease characterized by high levels of anti-nuclear antibodies and clinical symptoms restrained in flares of non-erosive polyarthritis. Six months prior, she developed hyporesponsiveness to recombinant human erythropoietin (rEPO) due to iron deficiency despite the administration of the appropriate dose of intravenous iron as well as the exclusion of other co-existing causes of resistance, namely inadequate hemodialysis, inflammation, malnutrition, severe hyperparathyroidism and underlying hematologic disorders. Surprisingly, the patient experienced a sudden episode of severe upper gastrointestinal bleeding characterized by hemodynamic instability, hemoglobin fall and melena that required blood transfusions. Once hemodynamic stabilization was achieved, an upper endoscopic exam was performed showing the pathognomonic extensive vascular ectasias and patchy erythematosus lesions at the distal antrum. Thus, GAVE should be considered in the differential diagnosis of chronic kidney disease patients who present not only with acute blood loss but also with resistance to rEPO due to inadequate iron stores.


Many cattle and sheep do not produce enough colostrum for their offspring.Prepartum nutrient intake is an accessible strategy for producers to influence colostrum production.Greater prepartum starch intake can influence colostrum composition and increase colostrum yield for beef cattle and ewes.Colostrogenesis is sensitive to fat intake, dependent on the dietary fatty acid composition: greater linoleic acid intake often increases colostrum antibody concentration.Colostral bioactive compounds are frequently altered by a prepartum diet without changes in overall colostrum composition. Prepartum nutrient intake could be strategically used to maximize beneficial compounds for the newborn.IgG), macronutrients, and bioactive compounds that promote physiological maturation and support the immune system  or protein (MP) intake, or dietary components such as starch, neutral detergent fiber (NDF), or fat.Most research evaluating how prepartum nutrient intake affects colostrum production has mainly focused on global nutrient under- or over-provision prior to parturition. Imposing mid to late-gestation nutrient restriction reduced first-milking colostrum yield  in beef cattle  without NE]). This is additionally complicated by using either empirical or mechanistic models to predict energy requirements and dietary sufficiency and which coefficients of energetic efficiency are used for the conversion of ME to NE for pregnancy or maintenance. Thus, utilization of either system can impose a bias on colostrum production responses between studies. Research evaluating carbohydrate consumption prior to parturition is shown in Carbohydrates proportionally contribute the greatest amount of dietary energy in ruminant rations and, as such, relative starch and NDF substitutions have consequences for dietary energy intake that often confound colostrum production responses. Furthermore, studies that manipulate dietary starch and NDF content to achieve specific energy intakes differ in which energy partition they target  and MP intake not always being balanced across treatments  increased colostrum yield by 80% in Simmental-Angus crossbred beef cattle . These cIn accordance with beef cattle, increasing dietary starch and energy intake in sheep consistently increased colostrum yield . Bancher1 and total IgG concentrations in colostrum ; thus, this factor should be considered when comparing colostral IgG concentrations between studies using different dietary starch sources. Nonetheless, IgG concentration in first-milking colostrum collected within 1 to 12 h after calving has been reduced by greater dietary starch inclusion or ME intake precalving  while suppressing the synthesis of de novo FA relative to cows fed a ration that targets 125% or 150% predicted ME requirements prior to calving  are substantial but transient and, as such, presumed negligible relative to pregnancy and maintenance requirements. Regardless, numerous studies .Models that evaluate the sufficiency of prepartum shown in  have evashown in  and 2. Wiso-nitrogenous and iso-energetic rations; Apart from substituting sunflower meal for cottonseed meal as a protein source wherein the authors observed that greater sunflower meal increases first-milking colostrum yield  increases colostrum yield . HoweverAs with colostrum yield, colostrum IgG concentration, as measured by radial immunodiffusion  or estimThere are limited data regarding how prepartum protein intake affects colostrum macronutrient composition and yield in cattle. That said, from what is available, colostral fat, protein, and lactose concentrations are unaffected , nor areComparatively, colostrum protein concentration has been reported to increase with protein supplementation in some studies with ewes , but notRations typically have 2%DM dietary fat and, generally, dietary fat should not exceed 6% to 7%DM to avoid detrimental impacts on ruminal fermentation. However, greater dietary fat provision can also improve ration palatability, control dust and fines, and alleviate heat stress due to its lesser heat increment respective to proteins and carbohydrates. Increasing prepartum dietary fat intake can concurrently increase energy intake without providing excessive dietary starch. Consequently, prepartum fat intake may have utility for prepartum rations and studies in cattle  and ewesLate-gestation fat inclusion rate and source have not been shown to affect first-milking colostrum production in dairy cattle, either when NE concentration was maintained  or when ɑ-linolenic) in late-gestation rations could be beneficial for increasing ewe colostrum yield. Currently, it is unclear why this response occurs.First-milking colostrum yield in ewes appears to be more responsive to dietary fat content than cattle, but results across studies remain inconsistent. Some authors have found that specific fat sources  can decrease first-milking half-udder colostrum yield collected within an hour  and betwInterestingly, colostral IgG concentration, as measured by ELISA or estimated using Brix, appears to be responsive to dietary fat inclusion and source in both dairy  and beefAlthough fewer data are available for ewes, their colostral IgG concentration does not appear to respond to fat intake and source as in cattle . Though,Colostral macronutrient concentrations are usually unaltered by dietary fat content and source for cattle . NotablyThe colostral FA profile is the most reported bioactive component among research evaluating prepartum dietary fat intake and source. For both cattle and sheep, colostrum FA composition ubiquitously varies across all reported studies when the dietary fat inclusion rate or source is altered in such a way that coincides with dietary FA intake and maternal plasma FA profile. With respect to other bioactive components and micronutrients, Prepartum nutrient intake has the capacity to influence colostrogenesis in bovine and ovine species and impact colostrum yield and composition. Although ewes are generally more sensitive to ration composition than cattle, there is evidence to suggest that prepartum nutrition can be used strategically to promote colostrum production in both species. Dietary starch and fat content and source appear to exert influence on colostrum IgG concentration, with data indicating that starch:NDF substitutions affects colostrum yield in beef but not dairy cattle. Current data suggest that cattle are irresponsive to dietary protein content, whereas ewe colostrum production responds to dietary protein source and intake. Further work needs to differentiate between carbohydrate, protein, and fat sources, accounting for total IgG and macronutrient yield, to characterize mammogenic capacity under differing nutritional regimens, particularly in relation to the feeding duration and timing of supplementation relative to the onset of colostrogenesis. In addition, there is a need to detail the colostral bioactive profiles, as this constituent often responds to prepartum nutrient intake without changes in the gross composition of colostrum. Colostral bioactive components are inherently important for neonatal development and, as such, prepartum nutrient intake might be used strategically to benefit the offspring.


Psychiatric disorders are complex clinical conditions with large heterogeneity and overlap in symptoms, genetic liability and brain imaging abnormalities. Building on a dimensional conceptualization of mental health, previous studies have reported genetic overlap between psychiatric disorders and population-level mental health, and between psychiatric disorders and brain functional connectivity. Here, in 30,701 participants aged 45–82 from the UK Biobank we map the genetic associations between self-reported mental health and resting-state fMRI-based measures of brain network function. Multivariate Omnibus Statistical Test revealed 10 genetic loci associated with population-level mental symptoms. Next, conjunctional FDR identified 23 shared genetic variants between these symptom profiles and fMRI-based brain network measures. Functional annotation implicated genes involved in brain structure and function, in particular related to synaptic processes such as axonal growth (e.g. NGFR and RHOA). These findings provide further genetic evidence of an association between brain function and mental health traits in the population.The online version contains supplementary material available at 10.1186/s12888-023-04905-7. Psychiatric disorders are complex, with a polygenic architecture, and large degree of overlapping symptoms and risk factors. Both imaging and genetics studies have shown numerous but small associations between brain phenotypes, psychiatric disorders, and genetics, such as schizophrenia (SCZ) –3, bipolWe have recently deployed a multivariate analysis to study the genetic architecture of brain functional connectivity, revealing genetic variants associated with functional brain connectivity as well as variance in brain activity over time . The resPrevious studies have shown widespread phenotypic and genetic overlap between psychiatric disorders –18. In aHere, we aimed to uncover the genetic architecture of mental symptoms and identify shared genetic loci with neurobiological processes related to brain function. Using multivariate analysis , we geneWe utilized data from the UK Biobank  with perThe processing pipeline for imaging data used for the multivariate GWAS is described in Roelfs et al. . In shorIn this study we applied MOSTest to the phenotypic data from the ICA decomposition described in Roelfs et al. . MOSTestIn order to quantify the degree of genetic overlap and identify shared genetic loci between the mental health profiles and the imaging features we deployed the pleiotropy-informed conjunctional false discovery rate (conjFDR) through the pleioFDR toolbox . We usedADH1B and ADH5 (chromosome 4) and CRHR1 (chromosome 17).MOSTest revealed 10 significant (P < 5e-8) loci across the 13 previously identified profiles of mental health  , bipolarNext, we explored the genetic overlap between the mental health profiles and the psychiatric disorders through the conjunctional false discovery rate (conjFDR) , 36 whicWe then calculated the number of shared genetic loci between the multivariate genome-wide association statistics for mental health profiles and the two multivariate measures of the brain functional connectome using conjFDR. The GWAS summary statistics from our previous study of brain function . In shorIn this study we identified a number of loci associated with multivariate genome-wide association statistics for mental health profiles and found overlapping loci with the measures of brain function and psychiatric disorders. Using MOSTest we were able to leverage the phenotypic overlap between different mental health profiles to identify new loci associated with a multivariate measure of mental health. Genes associated with these loci showed regional expression in different parts of the brain .Our analysis using conjFDR revealed a number of shared loci and genes between the multivariate genome-wide association statistics and the psychiatric disorders. This demonstrates the shared genetics between psychiatric symptoms regardless of clinical diagnosis, emphasizes the utility of using population-level phenotypes to investigate variance in mental health profiles, and highlights the advantage in leveraging pleiotropy between complex phenotypes to boost discovery. We found shared genes with all but two case-control GWAS , which also had the two smallest sample sizes, which may reflect insufficient power to detect an effect , or may We also identified a number of overlapping loci between mental health profiles and fMRI measures of brain function, including 18 shared loci with functional connectivity and 5 shared loci with node variance. The higher number of shared loci for functional connectivity might be partially explained by the number of phenotypes in each composite measure. While the functional connectivity GWAS comprises 210 measures, i.e. partial correlations between 21 brain nodes, the node variance GWAS encompasses only the temporal variance in each node. It is possible that the number of phenotypes included in the multivariate analysis can affect the discovery . Both thThe two conjunctional analyses with fMRI measures and the multivariate genome-wide association statistics for mental health each showed a number of overlapping loci. Not all shared loci were unique, this can be partially explained by the definition of the brain networks in our analyses. The functional connectivity and the node variance measures use the same 21 nodes, and, ultimately, the two measures reflect different properties of the same time series. We found that the number of overlapping loci between the multivariate genome-wide association statistics for mental health and the brain functional connectome was generally larger than findings from our previous study highlighting shared genetic loci between psychiatric disorders and the brain functional connectome. This may partly be due to the larger sample size of the multivariate measure of mental health, but it could also reflect that the multivariate genome-wide association statistics capture genetic variance more generally related to the brain functional connectome. We found little direct overlap between loci of these two GWAS’ separately, which highlights the discovery boost advantage of using conjFDR in phenotypes with generally low heritability.The main implication from our findings is that we can identify shared genetic variants between a multifactorial measure of mental health in an undiagnosed population sample and fMRI-based measures of brain functional connectivity. Several limitations should be considered. First, the data were obtained from a middle-aged and older White British population, which limits the generalizability of the findings. Further, the mental health questionnaires are self-administered, so the data is vulnerable to various response and self-selection biases . We exclIn conclusion, our multivariate GWAS on 13 mental health symptom profiles showed a number of shared genetic loci with two fMRI-measures reflecting brain function and connectivity. This research shows that genetic overlap between mental health symptom profiles and brain functional connectivity can be linked to, among other processes, axonal growth regulation (through NGFR and RHOA) and regulation of MECP2 transcription factors (through MEF2C). This provides further genetic evidence of an association between brain function and mental health traits in the population.Below is the link to the electronic supplementary material.Supplementary Material 1: Supplementary Figures and Tables


Editorial on the Research TopicMolecular pathogenesis and novel treatments for inherited cardiomyopathiesInherited cardiomyopathies have diverse genetic etiologies, but together are major contributors to cardiac disease that affect patients of all ages, typically with early onset in childhood or adolescence. While inherited cardiomyopathies are currently classified according to functional and morphologic features, finer resolution of these categories has been made possible with the aid of modern genetics. However, despite the identification of many disease-causing variants, effective therapies for the majority of inherited cardiomyopathies remains scarce. In this issue, a collection of original research and review articles unveil and summarize the pathogenic mechanisms and gene expression signatures of various inherited cardiomyopathies as well as novel and extant therapeutic strategies in treating selected subsets of inherited cardiomyopathies.Hilal et al. which discusses in detail how mutations in RAS genes underlie phenotypically diverse cardiomyopathies in addition to distinct systemic syndromology. Notably, one must cautiously distinguish cardiomyocyte-intrinsic from extrinsic effects of RAS hyperactivation. While several in vitro studies demonstrated that ectopic RAS activation results in cardiomyocyte hypertrophy, cardiomyocyte-specific expression of gain-of-function RAS variants did not result in HCM despite cardiomyocyte hypertrophy seen in neonatal mice carrying the same germline mutation in the RAS gene , represents a major co-morbidity of genetic hemochromatosis as well as secondary iron overload. A recent clinical trial demonstrated that amlodipine and chelation combination therapy significantly lowers intracardiomyocyte iron deposition. Mechanistically, amlodipine appears to block the L-type calcium channel, which otherwise provides a major route for iron entry into the cardiomyocyte. Over the past decades, the identification of genes in which pathogenic variants are associated with inherited cardiomyopathies has raised expectations for new therapies. Advances in genomic editing technology hold promise for directly targeting pathogenic variants, and have the potential for shifting the paradigm for treatment in genetic medicine. Other approaches harnessing natural genetic modifying mechanisms that suppress the penetrance of pathogenic variants may also be useful. While the incomplete penetrance of inherited cardiomyopathies often complicates the genetic evaluations of families with cardiomyopathies, the observation that many mutation carriers remain disease free in fact provides paradoxical hope that disease-modifying therapeutics may be achievable. Future omics studies comparing transcriptome and proteome between clinically active and silent mutation carriers may thus uncover novel therapeutic approaches to delaying disease onset. In parallel, development of additional genetic animal models that approximate human cardiomyopathies will enable rigorous mechanistic studies that potentiate development of therapeutic agents.


Structural valve deterioration is commonly defined as an intrinsic permanent change of the bioprosthesis due to leaflet calcification, thickening, pannus formation, tear, or disruption. The resulting deterioration leads to stenosis and/or intra-prosthetic regurgitation. Here we present the case of a 75-year-old patient who underwent aortic valve replacement with a bioprosthetic valve. Predischarge transthoracic echocardiography revealed an aortic prosthesis with normal gradients. Three years later, the patient reported exertional dyspnea. Transthoracic echocardiography was performed and showed a high transvalvular pressure gradient of 80 mmhg with restricted mobility of the leaflet caused by subprosthetic tissue. Redo aortic valve replacement was planned and surgical resection was performed showing pannus ingrowth in both aortic and ventricular side of the bioprosthesis.


We also do not understand how suicide and suicidal ideation changed for different Hispanic immigrant populations since 2015, coinciding with a rise in anti-immigrant political sentiment and policymaking. Without action to either improve public health surveillance or leverage existing data sources with immigration status information to better understand the burden of suicide death and suicide risks among Hispanic immigrants, we may fail to prevent unnecessary loss of life.,,Anti-immigrant attitudes are not a new phenomenon in the U.S.,A key example of these policies was the administration's highly publicized public charge rule changes, which built upon the public charge standard developed through the Illegal Immigration Reform and Immigrant Responsibility Act.,Unfavorable immigration policy has been linked to greater perceived health problems among adults and children.,Despite these risks, deportation fears and the chilling effects of anti-immigrant attitudes and policies likely discouraged enrollment into health insurance programs ,Avoiding care while facing discrimination or psychological trauma can be problematic for suicide prevention. Discrimination is a risk factor for mental health problems among Hispanic persons.In response, public health researchers have an opportunity to increase their understanding of the burden of suicide among Hispanic immigrant populations in relation to the rise of anti-immigrant sentiment. This remains a challenging task when national mental health and suicide statistics do not take immigration status into account, when commonly available data sources restrict access to information about visa status, and when data from the Centers for Disease Control and Prevention do not distinguish between immigrants and U.S.-born Hispanics.Many researchers have attempted to account for immigration status in their studies―cutting across multiple disciplines―although these attempts have been discordant, resulting in different definitions and experiences of unauthorized legal status.Specific actions can be taken while being mindful of stigma-related implementation concerns. First, rather than actively seeking information from Hispanic immigrants, we can prioritize efforts to report deceased immigrants’ citizenship status on either the U.S. Standard Certificate of Death or equivalent state-level forms. Specifically, citizenship status, available by contacting Citizenship and Immigration Services, could feasibly be collected by the medical certifier or funeral director when collecting demographic data required for the death certificate. This information would then be reported into state violent death report systems, which already aggregate deidentified information on country of birth and other sensitive characteristics for suicide decedents.Second, additional initiatives could include collecting citizenship information for immigrants participating in large health surveys. Previous studies suggest that unauthorized immigrants are interested in and willing to discuss legal status in research. To ease the impact of the deportation-related concerns on unauthorized immigrant participation, surveyors should build rapport before presenting legal status questionsThird, the research community can use existing data in creative ways to better understand suicide-related risks experienced by Hispanic immigrants. Data sources collecting citizenship or country of birth information, such as the National Health Interview Survey or Hispanic Community Health/Study of Latinos, respectively, can be used to estimate the relationships between immigration status and important suicide risk factors for Hispanic populations since 2015. Existing data sources can also give us guidance on how to estimate the impact of recent immigration policies on suicide risk factors such as mental health outcomes.We do not fully understand the burden of suicide or recent changes in suicidal ideation among Hispanic immigrant populations in the U.S. However, we hypothesize that increasingly acrimonious immigration attitudes and policies likely worsened suicide-related outcomes for many Hispanic immigrants, authorized and unauthorized. The coronavirus disease 2019 (COVID-19) pandemic is likely intensifying suicide-related risks, given the harmful impacts the pandemic has imposed on Hispanic communities.


The United States Department of Health and Human Services (HHS) pledged $90 million to help reduce health disparities with data-driven solutions. The funds are being distributed to 1400 community health centers, serving over 30 million Americans. Given these developments, our piece examines the reasons behind the delayed adoption of big data for healthcare equity, recent efforts embracing big data tools, and methods to maximize potential without overburdening physicians. We additionally propose a public database for anonymized patient data, introducing diverse metrics and equitable data collection strategies, providing valuable insights for policymakers and health systems to better serve communities. This sum is planned to be distributed to 1400 community health centers (CHCs) serving over 30 million Americans1. Specifically, this pledge will allow health centers to expand analytics and reporting capabilities to enhance healthcare services while supporting patient-level UDS+ data submissions to collect more precise data on health disparities2. We herein propose a national public database featuring voluntarily self-disclosed and deidentified patient data to increase the granularity of health disparities metrics, and propose how this project can incorporate diverse metrics and equitable data collection procedures.In April 2022, Health and Human Services (HHS) pledged 90 million USD to the American Rescue Plan Uniform Data System Patient-Level Submission (ARP-UDS+) funding award in support of new data-driven efforts for Health Resources and Services Administration (HRSA) Health Center Programs to identify and reduce health disparities3. The NHQR is a comprehensive review of American healthcare comparing health outcomes across both state and national levels4. This report revealed that, although communities of color have enjoyed increased healthcare access since 2000, racial inequity persisted in 2021 due to lack of focused interventions addressing disparities3. While CHCs disproportionately serve marginalized populations, their lack of granular data collection is likely leading to underestimations of health disparities in areas requiring the greatest resource support4.Big data has already been proposed to promote health equity at the federal level. For instance, in 2021, the Agency for Healthcare Research and Quality (AHRQ) released the National Healthcare Quality and Disparities Report (NHQR)4. For instance, the Community Health Needs Assessment (CHNA) in the Patient Protection and Affordable Care Act (ACA) requires CHCs to investigate the biggest health-related challenges in their communities and outline solutions5. However, CHNAs often lack subjective data like medical trust and are only collected periodically, producing an unnecessary lag time between when systemic health issues arise and when healthcare providers can identify and respond to them.Big data has impacted patient care for decades by helping health insurance companies incentivize preventative care among patients and physicians, ultimately decreasing the use of costly acute care and improving care equity4. This platform can also be used to monitor medications while empowering CHCs to pinpoint and respond to systemic issues more quickly. Although the database may be skewed towards higher-income individuals with more time to self-report, the resource can help compare assess healthcare quality without overburdening physicians, ultimately improving equity interventions.Given these challenges with current healthcare metrics, the Centers for Medicare and Medicaid Services (CMS) should develop a national database like the CDC’s Vaccine Adverse Event Reporting System (VAERS), which collects self-reported issues with vaccine reactions, to regularly collect self-reported data on symptoms, diseases, adverse reactions, medical trust, and perceptions of health equity through metrics like transportation time or food insecurity6. These metrics were only introduced to eight of these measurements, and that only in 20226. HEDIS also does not stratify metrics by other determinants of health like food insecurity or employment status6. Metrics for further development could also include language access, transportation barriers, and health literacy.Although the data would be self-reported, the requested data should include many new metrics. For instance, the National Committee for Quality Assurance (NCQA) Healthcare Effectiveness Data and Information Set (HEDIS), the basis for many equity measurement systems, HEDIS was inaugurated in 1991 and continued for 31 years without race or ethnicity included in its nearly one hundred performance measures7. CHCs can also assess the readability of patient education materials and track the effectiveness of health literacy interventions like counseling, visual aids, staff training on health literacy communication, and simplified language in educational pamphlets through validated health literacy screening tools like REALM-SF (Rapid Estimate of Adult Literacy in Medicine - Short Form)8. REALM-SF examines abilities to read aloud medical terminology and is available free for download on the National Center for Education Statistics (NCES) website9.Healthcare organizations can better understand language barriers by offering patient surveys after each visit. Depending on the extent of the issue, Community Health Centers (CHCs) can then adopt interpreter services and track its effectiveness through surveys. To address transportation barriers, patients can opt to disclose transportation service use and reasons for missed appointments. CHCs can then partner with transportation providers like Uber WAV (Wheelchair Accessible Vehicles) and Uber Health to improve patient accessibility and cover for deficiencies in available wheelchair-accessible vehicles11.To ensure that the system captures data from marginalized populations, we propose CHCs invest in technology infrastructure and staff training to prepare for more comprehensive data collection by, for instance, leveraging telehealth services to provide care management while streamlining data collection through remote patient monitoring and automated post-checkup surveys. Then, outside partnerships with research organizations can help CHCs analyze this data and devise tailored solutions through partnerships with relevant nonprofits like the National Partnership for Women and Families, which has developed best practices for patient engagement in healthcare, and Leapfrog, which helps analyze data to promote healthcare safety and qualityFurthermore, to promote interoperability with electronic health records, we suggest following the technical standards developed by organizations such as the Office of the National Coordinator for Health Information Technology (ONC) and the Health Level Seven International (HL7) organization. These standards ensure that data can be exchanged seamlessly between different healthcare systems while safeguarding self-reported data. We also recommend consulting with the ONC’s Trusted Exchange Framework and Common Agreement (TEFCA) to help establish policies for securely exchanging health information across organizations.However, marginalized groups must be involved in the design of this database. They could provide input on how to create a user-friendly interface with icons instead of words on buttons to accommodate those with low literacy or limited technological proficiency. The database should also collect data on the widest possible range of demographic factors; provide technical support through online tutorials or help desks staffed by technical support personnel; establish robust privacy policies like strict data access controls, encryption protocols, and other technical safeguards; and implement data quality controls, like automated data validation checks; and seamlessly deliver the data to researchers and policymakers through data dashboards.As big data revolutionizes the world, we must not forget its potential impact on improving health outcomes and disparities. By investing funds into health centers and hospitals for patient-driven reporting and data collection on race and ethnicity and developing minimum standards for health equity metrics collection and reporting across states, public discourse and media then draw on subsequent patient data analyses to bring greater attention to regional and national healthcare inequities, leading lawmakers to prioritize health equity initiatives and healthcare systems to have a clearer picture of the communities they serve.Further information on research design is available in the Reporting Summary


In recent years, the advancement in biotechnology has enabled much improvement in quality and outcomes of medicine. These innovations have also drawn great attention in dental research fields. Regenerative medicine such as tissue (bone and tooth) engineering has been a hot topic in oral and craniofacial research for a few decades. Cells, signaling molecules and scaffold materials are three key components of tissue engineering approaches. Almost everyday new bioengineering approaches are proposed and tested for various diseases and treatment in dentistry.Liu et al.). EVs secreted by human gingival MSCs (hGMSC-derived EVs) were shown to promote osteogenesis and neovascularization in vitro and in vivo . The roles of stem cell-derived EVs and non-stem cell-derived EVs in bone tissue regeneration  are reviewed . Engineering modified EVs may play important roles in future cell free EV-based bone tissue engineering therapies.Stem cells such as mesenchymal stem cells (MSCs) are multipotent and may differentiate into different cell types for tissue repair and regeneration. However technical issues including phenotype consistency, host immune response and potential tumorigenicity are still not completely resolved. Extracellular vesicles (EVs) originate from cellular endosomes and contain bioactive molecules to target cells by paracrine. It is known that these EVs are one of the major mediators of stem cells leading to their biological effects. MSC-derived EVs (MSC-EVs), instead of MSCs, may be potentially used in tissue repair and regeneration. The current status and future therapeutic applications of MCS-EVs in oral and craniofacial tissue regeneration are discussed in a review . Similarly, nanofibrous scaffold material can be modified to provide better coordinated regenerative endodontic treatment when pulp connective-tissue, dentin formation, revascularization and reinnervation need to be well orchestrated . Silk fibroin nanoparticles were coated with genetically engineered cell membrane overexpressing toll-like receptor 4 and loaded with minocycline hydrochloride. These biomimetic nanoparticles demonstrated excellent targeted antibacterial and immunoregulatory effects in vitro and in vivo (ligature-induced periodontitis mouse model) .A new generation of scaffold materials has been developed for tissue engineering. A multifunctional structurally optimized hydrogel scaffold was designed by integrating polyvinyl alcohol, gelatin, and sodium alginate with aspirin and nano-hydroxyapatite (nHAP). The osteogenic of nHAP and anti-inflammatory function of aspirin were successfully synergized . Targeted delivery of antitumor drugs has been recognized as a promising therapeutic modality to improve treatment efficacy, reduce toxic side effects and inhibit tumor recurrence. A controlled-release of an anti-tumor drug (apatinib) from supramolecular nanovalve-modified mesoporous silica was designed for targeted inhibition of osteosarcoma . This in vitro study showed efficient release of antitumor drug and promising results for future osteosarcoma treatment. Enamel white spot lesions do not have effective yet conservative treatment methods. Synergistic remineralization of enamel white spot lesions was achieved by using mesoporous bioactive glasses loaded with amorphous calcium phosphate, which implies great potential for clinical application .Other advancements in oral disease therapies are also made in recent years. Eldecalcitol, a novel active vitamin D3 analog, was shown to be effective on preventing alveolar bone loss in diabetes-associated periodontitis . Understanding the composition and differences between supragingival plaque biofilm microbes on the surface of fixed prostheses and natural crowns can provide patients with targeted guidance to focus on oral hygiene habits, reduce the risk of periodontal diseases and improve the success rate of fixed prostheses.The microbial composition and structural diversity of supragingival plaque on the surface of fixed prostheses were found to differ from that of normal natural crowns, with relatively high levels of periodontally related pathogens and higher microbial metabolism. The microflora on the surface of all-ceramic crowns was more similar to that of natural crowns than to that of porcelain-fused-to-metal crowns . Through the use of AM technology, personalized implant treatment for individual patients can be achieved.In addition to biotechnology, digital technology has also been applied in oral disease treatment, such as implant therapy. Additive manufacturing (AM) can enable the direct fabrication of customized physical objects with complex shapes, based on computer-aided designs. The applications of AM technologies in oral implantology, including implant surgery and restorative products, was reviewed (With the quick advancement of therapeutic approaches, biotechnology, and bioactive materials, it will not be too long before precision and personalized dentistry become reality.


Background: Older people living in residential aged care facilities frequently experience medicines-related harm. Evidence regarding the perception and practices towards reducing these harms may facilitate the development of customised educational programs for pharmacists providing services in RACFs.Objective: To explore Australian pharmacists’ opinions and practices towards reducing the risk of medicines-related harm in aged care residents.Methods: An online survey was developed based on a literature review, expert opinion, and feedback from pharmacists providing services in RACFs. A web link for the survey was shared via professional pharmacy organisations and social media groups with Australian pharmacists providing services in RACFs.Results: A total of 209 pharmacists participated in the survey. Of these, 76% (n = 158) were residential medication management review embedded pharmacists, and 24% (n = 51) were supply pharmacists for RACFs. Most pharmacists believed that medicines-related harm is common in residents , yet few agreed that pharmacists have enough time to participate in medicines-related harm reduction services . There was a high level of agreement regarding the key risk factors  and potential strategies  for reducing medicines-related harm in residents.Conclusion: Pharmacists agreed that older residents often experience medicines-related harm, but they did not frequently participate in medicines-related harm reduction services. Initiatives to engage pharmacists in team-based harm reduction services and educate aged care staff regarding safe medication management may improve residents’ safety and health outcomes. Medicines play an integral role in disease management, yet can be associated with significant problems, especially in vulnerable older people . Older pPharmacists are recognised as experts in pharmacotherapy and could potentially prevent medicines-related harm in older people . A pharmPrevious studies in Australian RACFs focused on determining pharmacists’ views towards antibiotic prescribing  and moniA cross-sectional national survey was conducted between February 2022 and August 2022. Pharmacists across Australia providing clinical or supply services to RACFs were the target population.An initial draft of the survey was generated based on a literature review, expert opinion, and feedback collected from pharmacists providing services to RACFs. The survey comprised questions on demographics, pharmacists’ extent of agreement with statements regarding medicines-related issues and practice considerations, and perceptions towards risk factors for medicines-related harm and potential strategies for reducing medicines-related harm in aged care residents. Question types included Likert scales and free-text boxes. The face and content validity of the draft questionnaire was determined through a pilot sample of 10 registered pharmacists. Based on feedback, the questionnaire was reviewed to ensure it was easy to understand and complete. It was designed to be completed within 15 min.® was shared with pharmacists via Australian pharmacy organisations , Pharmacy Daily (an online publication), and social media groups . An information sheet was on the cover page of the survey to provide general information to the potential study participant, including eligibility such as providing any type of services to RACFs. Completion of the survey was deemed as implied consent. We opted for the “Off” setting under multiple responses in SurveyMonkey® portal, which prevents the survey from being taken multiple times from the same device. Further, we added an additional note “If you have already taken this survey, then you do not need to submit it again” on the cover page of the survey to prevent multiple responses from the same pharmacist. All participants who completed the survey were entered in a draw to receive one of two AUD$100 gift cards. All returned questionnaires were reviewed for eligibility and completion.In 2020, the total number of RACFs in Australia was 3,300 . An assuU test for non-parametric numerical variables, were used to compare responses between the RMMR/embedded and supply pharmacists. Many supply pharmacists also provide services as RMMR pharmacist after receiving case-related training and accreditation from the Australian Association of Consultant Pharmacists. The purpose of comparing these groups was to assess the differences in their perceptions and practices towards reducing medicines-related harm among aged care residents. A p-value of <0.05 was used as the level of significance in all analyses.Data analysis was carried out using SPSS  and Microsoft Office Excel 2019. The mean ± standard deviation was used to present normally distributed continuous data. Ordinal or skewed data were presented using the median [interquartile range (IQR)], and frequency (percentage) was used to report categorical variables. The study questionnaire was comprised of four main sections: Perception (13 items), Practices (6 items), Risk Factors (11 items) and Strategies (7 items), and a 5-point Likert scale (1 = Strongly disagree to 5 = Strongly agree) was used. Inferential statistics, such as a chi-square test for categorical and Mann-Whitney Approval was obtained from the Tasmanian Human Research Ethics Committee (Reference: H0026755). We followed the reporting guidelines of the STROBE  statement for observational studies .From February to August 2022, 209 pharmacists completed the survey. One hundred and forty-six were RMMR pharmacists and twelve were embedded pharmacists within RACFs; we combined these groups as their roles are relatively similar and distinct from supply pharmacists. The demographics of the pharmacists are detailed in n = 174, 83%) and only half believed that safe medication management is usually practiced in RACFs . Almost all pharmacists agreed regarding the risk factors for medicines-related harm in aged care residents, such as polypharmacy, PIMs, dementia, recent drug changes, shortage of aged care staff, transitions of care, renal impairment, and specific drug use . A minority of respondents agreed that antibiotics  and antipsychotics  are usually prescribed in appropriate circumstances in older residents.n = 199, 95%). Most pharmacists also agreed that RMMRs prevent medicines-related harm , although only 62% of the RMMR/embedded pharmacists believed that their recommendations during RMMRs were typically accepted by general practitioners. Most pharmacists, particularly within the RMMR/embedded pharmacist group, agreed that aged care management personnel consider pharmacists’ suggestions for reducing medicines-related harm in residents , but few agreed that pharmacists have enough time to participate in medicines-related harm reduction services .As with risk factors, the pharmacists displayed high agreement regarding potential strategies  for reducing medicines-related harm in aged care residents. Enhancing collaboration between pharmacists and aged care staff was acknowledged by almost all pharmacists as a key strategy to reduce this harm in older residents ; this was lower for supply pharmacists (29%) relative to the RMMR/embedded pharmacists (60%). Also, a significantly lower proportion of the supply pharmacists reported that they were familiar with medication appropriateness tools  and routinely utilised these tools. Overall, very few supply pharmacists participated in harm reduction services, reported adverse drug reactions to the Australian Therapeutic Goods Administration, and utilised tools to predict the risk of medicines-related harm. There was a large difference in the proportions of supply and RMMR/embedded pharmacists  who agreed that supply pharmacists detect medicines-related harms whilst supplying medicines. Most pharmacists indicated that they routinely recommend deprescribing interventions ; this was lower for supply pharmacists (47%) relative to the RMMR/embedded pharmacists (92%).Only half of the pharmacists indicated that they routinely participate in harm reduction services (The role of pharmacists in aged care is evolving globally and may be critical in reducing the incidence of medicines-related injury in aged care residents . KnowingMost pharmacists agreed that medicines-related harm is highly prevalent in residents. Aged care residents take many medicines for their multiple morbidities, and it is a responsibility of all stakeholders involved in managing older residents to ensure the safe medication management . All staPharmacists believed that older residents frequently experience medicines-related harm due to factors such as polypharmacy, PIMs, anticholinergic drug use and multi-morbidity. They believed increased collaboration between pharmacists and aged care staff would reduce these harms. The 2021 Australian Aged Care Royal Commission report recommended increased communication and collaboration between aged care staff and other healthcare professionals, such as general practitioners and pharmacists providing services in RACFs . In New Relatively few pharmacists in our study agreed that antibiotics and antipsychotics are frequently prescribed in appropriate circumstances in Australian aged care residents. The RedUSe intervention, comprising multidisciplinary case review, provision of education to the staff, and audit and feedback, has recently been introduced in RACFs to reduce inappropriate antipsychotic prescribing . AntibioMost RMMR/embedded pharmacists believed that supply pharmacists do not usually identify medicines-related injury whilst supplying medicines. Many supply pharmacists in our study were not familiar with tools, such as the Beers criteria and anticholinergic drug burden scale, to predict the risk of medicines-related harm in aged care residents. In addition, very few supply pharmacists regularly utilised these tools or routinely engaged in harm reduction services. Similarly, a Malaysian study reported that only 27% of community pharmacists were familiar with Beers criteria and 17% frequently used this tool in practice . Educativia supplying medicines and consultancy, and these roles are executed remotely or in a visiting capacity (In Australia, pharmacists usually provide services in RACFs capacity . There icapacity . Embeddecapacity . In 2023capacity . A recencapacity .This study is the first of its kind that explored the perceptions and practices of Australian pharmacists providing services in RACFs towards reducing the risk of medicines-related injury in residents. One of the key study limitations was that the recommended sample size was not reached, indicating that these findings may not be representative of the entire group of Australian pharmacists providing services for aged care residents.Pharmacists agreed that aged care residents frequently experience medicines-related harm, but their involvement in medicines-related harm reduction services was relatively limited. Initiatives to engage pharmacists in harm reduction services and educate aged care staff regarding safe medication management may improve residents’ health outcomes. Medicines use in aged care residents can be optimised by the presence of embedded pharmacists in a team environment, provision of education to aged care personnel, and collaborative medication reviews with general practitioners. Ideally, supply pharmacists would also play a greater role in accepting responsibility for the risks posed by the medicines they provide to RACFs. Future research is needed to assess the impact of educating aged care staff about safe medication management on residents’ health outcomes.


Elevated peripheral inflammation is common in psychosis. Impairments in general cognition were linked to elevated C-reactive protein (CRP) and other inflammatory markers in patients with psychotic disorders. Whether there is a subgroup of persons with elevated peripheral inflammation demonstrating deficits in specific cognitive domains remains unclear. While molecular underpinnings of altered inflammation in psychosis are hypothesized, genetic contributions to relationships of psychosis, inflammation, and cognition have not been clarified. Thirteen peripheral inflammatory markers and 17 neurobehavioral tasks were quantified in a subset of participants  from the Bipolar-Schizophrenia Network on Intermediate Phenotypes (B-SNIP) consortium. Principal component analysis resulted in 5 inflammation factors across inflammatory markers. Three latent cognitive domains  were characterized based on the neurobehavioral battery. Hierarchical clustering identified a psychosis subgroup with elevated inflammation and worse cognitive performance. Genetic predispositions to schizophrenia and cognition were explored in relation to inflammation. Among persons with psychosis, higher inflammation indices were associated with impairments of Inhibitory Control and Visual Sensorimotor function. Greater deficits in Inhibitory Control were observed in a high inflammation patient subgroup. Consistent with previous studies, global genetic correlations of schizophrenia, CRP, and cognition were observed. Significant bivariate local genetic correlations of CRP with schizophrenia or cognition across 22 loci with several genes in 1 locus on chromosome 3 suggested pleiotropic mechanisms for inflammatory relationships with cognition and psychosis. Specific neurobehavioral domains may be more sensitive to inflammation dysregulation in psychosis as compared to general cognitive function, particularly performance on tasks requiring ongoing behavioral monitoring and control. These, along with evidence of genetic correlations of CRP, psychosis, and cognition, provide further supporting evidence that inflammation dysregulation is an important underlying mechanistic contributor to the disruption of cognition in psychosis. Targeting this dysregulation may be an avenue for novel therapeutics to improve cognitive outcomes in these patients.None Declared


Migraine is a primary headache disorder recognized by the World Health Organization as one of the most poorly understood and debilitating neurological conditions impacting global disability. Chronic pain disorders are more frequently diagnosed among cisgender women than men, suggesting that female sex hormones could be responsible for mediating chronic pain, including migraine and/or that androgens can be protective. This review discusses the major gonadal hormones, estrogens, progesterone, and testosterone in the context of molecular mechanisms by which they play a role in migraine pathophysiology. In addition, the literature to date describing roles of minor sex hormones including prolactin, luteinizing hormone, follicular stimulating hormone, and gonadotropin releasing hormone in migraine are presented. Because transgender and gender non-conforming (trans*) individuals are an underserved patient population in which gender-affirming sex hormone replacement therapy (HRT) is often medically necessary to align biological sex with gender identity, results from cisgender patient populations are discussed in the context of these major and minor sex hormones on migraine incidence and management in trans* patients. Migraine headaches are diagnosed based upon attack frequency and aura. When headache symptoms are present for at least three months, episodic migraine is defined as 0–14 headache days per month while chronic migraine requires 15 or more headache days . The phy1.2.Cortical spreading depression is defined as the spreading of a slowly propagating wave of cellular depolarization and inhibition within neuronal and glial cells across the cerebral cortex. A large change in the concentration gradient between extracellular potassium and intracellular sodium/calcium ions produces a depression of electrocortical signals that travels anteriorly at a rate of 3–6 mm/min , 6. In aPreliminary research on the possible involvement of CGRP in migraine found that patients experienced elevated CGRP levels during the interictal phase when headache symptoms were absent ; furtherGiven that trigeminovascular neurons have intrinsic projections to the cerebrovascular system, cortical hyperexcitability induced within central neurons activates and amplifies the underlying vascular processes that sustain migraine headache. The current neurovascular understanding of migraine suggests that neurological processes are necessary to alter trigeminal activity . Clinica1.3.Migraine is more commonly diagnosed with a higher rate of disability in cisgender  women than men , 21. AltResearch on sex differences usually refers to the hormonal differences between female sex hormones  and male sex hormones , but the correlation between migraine and menstrual sex hormone fluctuations reveals that the physiological concentration as well as the type of sex hormone is critical for headache pain. The human menstrual cycle typically lasts 28 days and is divided into two stages: the follicular and luteal phase. In the follicular phase, progesterone levels remain low while estradiol steadily increases. Ovulation then triggers a sharp decline in estradiol simultaneous with rising progesterone levels; when progesterone peaks halfway through the luteal phase, estradiol remains slightly elevated until both hormones drop back down to baseline , Figure Sex hormones are lipid signaling messengers that can enact long-lasting effects on neurovascular structure and function. To provide more comprehensive treatment for cisgender women/men and to expand the much-needed area of research on chronic conditions within transgender/gender non-conforming (shortened to trans* as a gender-inclusive umbrella term) patients who receive gender-affirming sex hormone replacement therapy (HRT), a comprehensive analysis on the impact of hormonal fluctuations in migraine-related pathological processes and on nociceptive processing is required. This review will first examine pre-existing, chronic pain-related health disparities among trans* patients to determine current healthcare needs regarding pain management and migraine headache. The medical necessity of gender-affirming treatment and current barriers to healthcare access will then be integrated to emphasize that gender affirmation is critical to reestablish important health measures—such as patient self-efficacy and trust in the healthcare system—that will grant trans* patients with the necessary skills needed to improve pain outcomes. Due to the significant knowledge gap between gender-affirming care and physical health disparities, research on sex hormone activity in migraine pathology will be summarized through cisgender patient data as it pertains to ovarian HRT (estrogen/progesterone) and androgen HRT (testosterone). Further gaps in knowledge and future research directions will be identified as needed to improve healthcare practice and to continue gender-affirming healthcare for LGBTQ + patients.2.2.1.When explicitly examining sex differences in primary headache, cisgender women report a higher migraine prevalence of 18% compared to 6% among men , whereas2.2.Estrogen is categorized as a “female” sex hormone synthesized within the ovaries to induce female sexual maturity; however, this simplification fails to consider the role of estrogen in male physiology as well as on other biological systems in cisgender women. Estrogen incorporates three hormones that have different functions based on the stage of female reproduction: estradiol is the main hormone that is sustained over a woman's lifespan while estrone and estriol are weaker forms that become dominant during menopause and pregnancy, respectively . EstrogeThe high prevalence of disabling migraine features among cisgender women implies that fluctuating levels of estrogen are critical for migraine development, but assumptions based on female physiology and the effects of hormonal signaling currently restrain the understanding of migraine to a system primed for estrogen. The difference in prevalence between migraine with aura during high estrogen and migraine without aura during estrogen withdrawal suggests that in addition to promoting migraine mechanisms, higher levels of estrogen increase CSD susceptibility and encourage significant headache-related disability through neurovascular dysfunction. Headache studies conducted using animal models demonstrated that female mice have intrinsically lower CSD thresholds than male mice , but wheGiven that CSD could be an initiating mechanism of migraine in patients, increased CSD susceptibility from female hormones indicates that estrogen has a critical role in triggering migraine wherein higher hormonal concentrations correlate to worse pain outcomes. However, clinical data from both cisgender and trans* patients differentiate the effectiveness of estrogen on male and female physiology. The introduction of estradiol to male physiology enables rapid CSD and central sensitization induction since low levels of pre-existing estrogen are insufficient to dominate testosterone activity, but for patients with female physiology, the developmental role of estrogen for sexual reproduction could require hormones that exceed healthy estradiol concentrations to propagate migraine mechanisms. During Tanner stage IV which occurs after puberty onset, patients with female physiology report estradiol levels between 15 and 85 pg/ml while male patients report a maximum of 38 pg/ml. In adulthood, female patients experience fluctuating levels of estradiol within the range of 15–350 pg/ml that varies according to the menstrual cycle ; during B receptors from the G-protein-coupled receptor attached to potassium ion channels; this suppression of neuronal hyperpolarization lowers activation thresholds because hyperpolarization is necessary to return neuronal activity back to baseline  pathway contains several metabolites vital to regulating excitatory and inhibitory signaling in the central nervous system. Kynurenic acid (KYNA) is a competitive antagonist that binds to the glycine site of the GluN1 subunit of NMDA receptors while other metabolites—such as quinolinic acid (QUINA) and xanthurenic acid (XA)—increase cortical excitability as orthosteric agonists that bind to the GluN2A-D subunit of NMDA receptors. QUINA and XA also activate type-2 metabotropic glutamate receptors . MigrainCentral sensitization consists of two critical components: cortical hyperexcitability facilitated through increased glutamate expression and enhanced synaptic efficiency through long-term potentiation; therefore, revealing the contributions of estrogen signaling on both aspects will improve headache pain relief. Clinical studies on migraine sex differences reported that in addition to elevated CGRP, cisgender women also experience fluctuating CGRP expressions within the central and peripheral nervous systems that correspond to estradiol concentrations across the menstrual cycle . EvidencEstradiol signaling seems to have a minimal role in directly perpetuating cerebral vasodilation since a significant portion of endothelial dysfunction and oxidative stress is proportional to the sensitized strength of trigeminal neurotransmission. However, research on the vascular contributions to migraine with sex hormones is still needed to elucidate the full impact of estrogen. In a study that compared plasma concentrations of nitric oxide among cisgender women with either menstrual migraine, non-menstrual migraine, or non-headache controls, enhanced activation of the nitric oxide pathway during the luteal phase leads to increased nitric oxide synthesis that aligns with reports of oxidative stress markers peaking during the late follicular and early luteal phases . EstrogeEstrogen-mediated disruptions in essential biomolecules bolster the efficiency of trigeminal neurotransmission at the cost of regulatory processes that are necessary to prevent excessive cortical hyperexcitability and cerebral vasodilation; therefore, since migraineurs demonstrate higher levels of serotonin (5-HT) during the ictal phase of headache but lower levels during the interictal phase, estrogen could influence the synthesis of neurotransmitters like serotonin . AlthougIn addition to generating intermediate metabolites that destabilize excitatory and inhibitory cortical signaling, the KYN pathway also reduces serum 5-HT levels in migraineurs by metabolizing the amino acid tryptophan into KYN metabolites at the expense of serotonin synthesis. Michael et al.  demonstrAn overview of the current understanding of estrogen signaling on neurovascular migraine mechanisms is summarized in 2.3.Although estrogen hormones are incredibly efficient at accumulating dysfunction within the trigeminovascular system, the role of fluctuating progestin hormones with estradiol signaling could clarify gaps in understanding between migraine headache, hormonal states, and pain symptomology. Progesterone has two receptors: progesterone receptor alpha (PRα) and progesterone receptor beta (PRβ). Although murine models demonstrate higher expressions of one receptor isoform over the other, human cells contain equivalent amounts of PRα and PRβ, which suggests that the PRα-PRβ heterodimer is the dominant molecular species for humans . Therefo2.4A receptor activity within neurons of the trigeminal nucleus caudalis  gene on chromosome 5 encodes three different isoforms: long (PRL-L), intermediate, and short (PRL-S), but only PRL-L and PRL-S will be considered since unlike humans and rats, mice lack intermediate PRL receptors . PRL-L ae models .Prolactin-induced sensitization imitates estradiol neurogenic inflammation by proliferating CGRP release from sensory neurons via the excessive activation of sensitized TRP channels. Although sensory neurons lack PRL, application of TRP channel activators to cultured female sensory neurons pretreated with prolactin resulted in amplified trigeminal neurotransmission due to enhanced calcium ion influx. Within male physiology, prolactin sensitization might require abnormally high concentrations to further support estrogen-mediated sensitization since males have lower PRL-L and PRL-S expressions in the dorsal root ganglion . PRL-S sWhen prolactin is considered alongside androgen and ovarian hormones, pronociceptive headache states fueled by estrogen can recruit prolactin-induced sensitization to further advance chronic pain since estrogen increases PRL receptor expression. Conversely, testosterone suppresses PRL expression to mitigate central sensitization, but since the molecular pathway defining this androgen-mediated phenomenon has yet to be explored, only coinciding interactions between estrogen and prolactin will be discussed. High concentrations of endogenous estrogen within female migraineurs correlate to an increase in PRL receptor mRNA in dorsal root ganglion neurons; however, because equal expressions of PRL-S and PRL-L were retained in both sexes regardless of estradiol concentrations, estrogen upregulation of PRL expression occurs independently from receptor isoform translation . ApplicaCRE mice to deduce sex differences in migraine. Prior to experimentation, murine models expressed KORCRE neurons in the dorsomedial/ventromedial hypothalamic nuclei and ARC as well as within tyrosine-hydroxylase-positive cells that were quantified as 80% for female and 70% for male mice. In response to RS, enhanced serum prolactin produced allodynia from latent sensitization in both male and female mice, although females experienced an intrinsically greater increase. When the KOR antagonist nor-binaltorphimine dihydrochloride was applied to the right ARC, serum prolactin was reduced likely due to the synchronized TIDA network  axis and reduced efficacy of the kappa-opioid receptor (KOR) system to alleviate trigeminal nociception. Previous clinical studies reported that stress activates the hypothalamus of migraineurs to increase cortisol levels, resulting in a positive correlation between stress-induced priming and chronic headache frequency. Since stress increases prolactin and the endogenous KOR agonist dynorphin, central sensitization could decrease the analgesic efficiency of the KOR signaling pathway such that chronic stress would hinder endogenous recovery from headache . Watanab network . Combinehibition , 81. Furhibition . Howeverhibition . Therefohibition . The mec2.6.2.Luteinizing (LH) and follicle-stimulating (FSH) hormones are synthesized from gonadotropin cells in the anterior pituitary to amplify estrogen synthesis during the menstrual cycle . Several2.6.3.Although the majority of gender-affirming healthcare focuses on medical interventions for trans* adults, trans* youth who wish to postpone the puberty of their assigned sex might receive puberty blockers—which are gonadotropin-releasing hormone (GnRH) agonists such as goserelin, leuprolide, and histrelin—to delay the development of secondary sex characteristics and prevent gender dysphoria—which is defined in the DSM-5 as the “psychological distress that results from an incongruence between one's sex assigned at birth and one's gender identity” , 89. SexGnRH is synthesized within hypothalamic neurons to regulate LH and FSH concentrations throughout the menstrual cycle; the activation of GnRH G-protein-coupled type I receptors on gonadotrophin cells in the anterior pituitary generates GnRH pulse frequencies wherein low pulses stimulate FSH and high pulses promote LH. Although female migraineurs demonstrated normal FSH/LH levels, dysfunction of the hypothalamic-pituitary-adrenal axis could cause abnormal GnRH pulse frequencies. Research demonstrated that persistent GnRH stimulation produces elevated LH:FSH ratios—amplifying the ovarian synthesis of androgen hormones—while minimal stimulation and abnormal GnRH serum levels were associated with hypothalamic amenorrhea . Despite3.Despite the growing number of trans* patients, healthcare practices have yet to move beyond the perspective that this patient population only engages with the healthcare system to receive gender-affirming medical treatment. Research examining pre-existing health disparities among LGBTQ + patients has been recognized as a crucial gap in knowledge ; howeverWhile the exact number of trans* individuals with migraine is unknown, there are an estimated 1.4 million transgender adults  and 39 million individuals with migraine in the United States . Based oAlthough this risk of headache could be detrimental for the physical and mental wellbeing of an already vulnerable population, transgender men and gender non-conforming individuals who receive androgen HRT might experience some relief from migraine. Yalinay Dikmen et al.  investigFrom the available evidence, estrogen HRT is associated with an increased risk for acquired migraine in transgender women while testosterone HRT could potentially provide some pain relief for transgender men with a pre-existing primary headache disorder. Although data from cisgender migraineurs has supported the role of estrogen in promoting migraine mechanisms, gaps in knowledge remain on the minimum physiological concentration of estrogen required to propagate chronic pain states; given that migraine has been observed during states of estrogen withdrawal  and elevation , determining physiological hormone concentrations can clarify the association of sex hormones and migraine symptoms. Consenting trans* patients represent the ideal clinical population to investigate this theory as the natural diversity in gender identity and use of gender-affirming hormones allows researchers to clarify the issue of sex hormone functionality without subjecting cisgender patients to potential unwanted side effects. These individuals are also ideal in furthering progress for LGBTQ + health outcomes beyond mental health disorders and sexual diseases because current debates regarding the legality of gender-affirming care already establish the immediate need for improved LGBTQ + healthcare and better understanding on the long-term effects of HRT. Treatment of headache for cisgender patients can also be improved through trans* data by reframing the consideration of health with gender identity; notably, by replacing binary sex as a health factor with measures of serum sex hormone levels, mechanisms involved in chronic pain pathology can improve the accuracy of disease diagnosis and treatment as well as support pain outcomes hindered by sexist gender stereotypes.4.Despite limited data in trans* individuals with migraine, cisgender data suggests that femininizing estradiol HRT increases the risk for acquired headache and chronic pain while masculinizing testosterone HRT is associated with improved pain relief for patients with a pre-existing headache disorder. Future studies are needed to directly evaluate the risk between gender-affirming HRT and migraine. Ovarian hormones like estrogen and progesterone promote migraine headache via CSD and central sensitization within the trigeminal nociceptive network through reactive species, CGRP, and an imbalance of excitatory/inhibitory neurotransmission while androgens like testosterone attempt to reduce excessive cortical hyperexcitability by bolstering neuroprotective and analgesic qualities. When the effects of sex hormones in cisgender and trans* patients are examined together, rapid sex hormone fluctuations create vulnerabilities through which heightened estrogen induces pronociceptive states and reduced testosterone fails to adequately establish trigeminal neuroprotection. Since many trans* individuals begin HRT when patients are already well into adulthood, pubertal hormones likely establish migraine susceptibility that can be aggravated further by estrogen HRT, thus putting transgender women at the most risk. Susceptibility could be curtailed through pubertal suppression and treatment with puberty blockers, but the same limitations currently impacting adult gender-affirming care also affect trans* youth, maybe even more so due to debates on parental rights to oversee the healthcare decisions of their children. Given the changing landscape of trans* patients seeking gender-affirming healthcare, more investigation into sex hormones on migraine pathology is warranted to determine the impact of gender-affirming HRT and potential chronic pain risk.Research on LGBTQ + healthcare has largely focused on educating medical providers about the appropriate terminology to improve interpersonal interactions between patients and providers , 99. How


Antibiotic resistance stands as one of the most significant public health challenges in recent decades. FEM proteins are responsible for the synthesisof pentaglycine cross-bridge, a primary constituent of bacterial peptidoglycan polymer crosslinking during cell wall biosynthesis. Since they are necessaryfor bacterial survival and antibiotic resistance, they were considered as significant antibacterial targets. We report herein, the virtual screening andselection of FDA-approved drugs and their potent similar molecules as FEM protein inhibitors and analyzed for inhibiting affinity and their ADMET pharmacokineticproperties. This data provide a foundation for the development and optimization of structurally innovative antimicrobial drugs. Staphylococcus aureus causes a wide spectrum of common ailments that can be acquired from the community and in hospitals. Due tolower responsiveness to commercially available drugs, the rise of Methicillin-resistant Staphylococcus aureus (MRSA) clinical isolateshas increased and evolved into various multi-drug resistant strains making the treatment difficult. The foreign expression of altered PBP2a performs the host PBPfunction and governs the strategy of β-lactam antibiotic resistance in methicillin-resistant S. aureus (MRSA)amino]-3-methylpyridin-2-yl]benzoic acid (PubChem ID: 90080139) against FemX with highest binding energy of -137.66 among all by forming a Pi-sulfur and Pi-alkylinteraction with TYR320, 377 and MET196. The whole complex has involved many hydrogen bonding with several residues listed in active site amino acids of theFemX receptor .In addiinvitro evaluations.For the first time this manuscript reported the FemB inhibitors from FDA approved drug molecules, which can be possibly repurposed.An overview of selected chemical molecules and clinical features with pharmacokinetic properties were analyzed using the pkCSM webserver and labeled ininvitro results. In this work, we virtually examined a library of FDA-authorized drugs from ZINC15 and DrugBank databases againstS. aureus FEM proteins. Three lead molecules were shortlisted based on their docking scores and prioritized based on their minimum bindingenergy and active interactions with the FEM proteins. In this study, we also evaluated the molecules solubility and toxicity properties.The limited therapeutic spectrum against MRSA necessitates an urgent development of antibiotics and medications to enhance the current antibiotic efficacyand drugs to enhance the existing antibiotic efficacy. We report the optimal binding features of structure-based FDA-approved drugs repurposed to treat MRSApathogenic infections. It helps to cross the emergence by following certain time limit management in collaboration with certain existingThe authors declare that they have no known competing interests (or) personal relationships that could have appeared to influence the work reported in thismanuscript. The research leading to these results received funding from the DST-SERB-ECR grant under file no: ECR/2017/003381.