FASEB BioAdvances

Background Training in grant proposal writing is an essential component of professional development for academic scientists in the biomedical and behavioral sciences. Despite the expansion of inter- and intra-institutional grant writing coaching groups as an approach to honing these skills, specific features that enhance or limit coaching group effectiveness have not been rigorously studied. Methods Qualitative inerviews were conducted with a subset of early-career investigators (n=204 and coaches (n=36) engaged in a national, U.S. based, group-randomized trial of grant writing coaching groups to test the effects of two variables on submission and funding of national-level proposals: (1) coaching duration (regular/extended dose) and (2) mode of engaging a scientific advisor (someone with content-aligned expertise) in the coaching process. This report focuses on interviews conducted upon completion of the regular coaching dose - 5 months of biweekly, group-based coaching sessions to support active proposal writing. Interviews were designed to identify which coaching group elements were perceived to be the most critical. Transcribed interviews were analyzed using deductive (participants) or open (coaches) coding to identify themes. Results Triangulation of results from participant and coach interviews showed strong concordance that coaches, peers, and scientific advisors all played key roles in supporting intervention efficacy. Sufficient alignment of scientific fields and/or methodologies among group members was important, although breadth of perspectives was also valued. Other critical group features were skilled and well-organized coaches, detailed feedback, peer-to-peer support (technical and psychosocial), and clear expectations for group functioning. Factors attenuating impact included variation in participants' engagement and "readiness to write," within-group mismatches of expertise or grant mechanisms, and limited research support at some participants' home institutions. Conclusion This study identified key elements of successful grant writing coaching groups and potential barriers to their effectiveness, while yielding insights about tailoring this approach for individuals at different stages of proposal development.

The iTHRIV Scholars Mentored Career Development Program initiated an Internal Grant Review (IGR) program in 2020 for current and recently graduated Scholars seeking funding through K and R awards from the NIH. The IGR program is designed to replicate the NIH review process and provide Scholars the opportunity to receive valuable feedback on their applications prior to NIH submission. A key characteristic of the program is the integration of REDCap, enabling automation and year-round offering while improving tracking and reporting efforts and capabilities. Results of the program have been overall positive, both in proposal development and participant feedback. IGR is a sustainable, valuable resource for early-career faculty competing for limited resources in the pursuit to become independently funded clinical translational scientists.

Funding and publishing scientific research has become increasingly competitive, highlighting a growing need for clear and accurate scientific communication. Scientific editors are professionals who help ensure that grant proposals and research manuscripts meet the standards of funding agencies and peer-reviewed journals while effectively communicating research significance to diverse audiences. However, detailed information characterizing the scientific editing profession is scarce. While some organizations have conducted salary surveys, to our knowledge, no comprehensive survey of the scientific editing field has been conducted. Here, we report findings from a 2024 comprehensive survey of 132 scientific editors. We collected data on demographics and education, career experience and current employment, skills and types of editing, and outcomes and satisfaction. Our data reveal that scientific editors have homogenous demographic characteristics but considerable variability in job titles, employment mechanisms, and career pathways. As the responsibilities of many scientific editors, including supporting proposal development, applying best practices in grantsmanship, and identifying strategies to enhance competitiveness for research funding, overlap with those of research development professionals, our findings provide considerations for further incorporating or defining the role of scientific editors within research development offices. Our results also suggest avenues for scientific editors to start or advance their careers. We anticipate that our results will serve as a foundation for discussions on the future directions of the scientific editing profession as well as areas for future research and professional development.

PurposeDiscussions surrounding the biomedical faculty job market often focus on applicant competitiveness and external metrics such as number of publications and funding records. Consequently, there is typically less discussion about applicant readiness, the point at which applicants perceive themselves as prepared to enter the market. Since 2018 our group, the Faculty Job Market Collaboration (FJMC), has conducted annual end-of-cycle surveys of biomedical faculty job applicants, producing the largest longitudinal dataset on this process to date. MethodsWe employed a mixed-methods design examining faculty applicants in biological science fields in North America. Regression analyses were conducted on a longitudinal dataset of 729 respondents across multiple hiring cycles. To determine how applicants evaluated their own preparation, qualitative interviews were conducted with a separate cohort of biomedical postdoctoral applicants during the 2024-2026 job cycles. ResultsOur findings demonstrate that rather than depending on a single quantitative threshold, readiness is a multifaceted construct shaped by actionable and interpersonal drivers. Key factors influencing an applicants perceived readiness include taking agency to submit applications, receiving explicit support from a mentor, incorporating strategic use of artificial intelligence tools into application preparation, and their career stage. ConclusionBy distinguishing individual readiness from systemic assumptions of market competitiveness, this study highlights a blind spot in academic workforce development. Our results suggest that applicants can achieve readiness and successful outcomes through different combinations of support, strategy, and timing rather than a uniform metric profile. By integrating quantitative and qualitative data, our study provides an evidence-based framework for understanding applicant readiness and offers practical guidance to help trainees navigate the increasingly competitive academic job market. Teaser TextOur mixed-model analysis of the biomedical faculty job market is designed to help prospective faculty candidates assess their readiness to enter the job market. By integrating multiple indicators of academic productivity, funding success, and professional experience, our study provides evidence-based benchmarks that can guide applicants in evaluating their competitiveness and identifying areas for further development before pursuing faculty positions.

IntroductionThe acquisition of National Institutes of Health (NIH) grants is an increasingly competitive undertaking. Many early-career biomedical investigators lack sufficient grant writing skills. This pragmatic, 2x2 factorial, group-randomized trial tested the effects of coaching duration and mode of engagement of participants scientific mentors in a grant writing coaching group intervention on grant application outcomes. MethodsA national US sample of 367 faculty and postdoctoral fellows, organized into 76 coaching groups across 6 cohorts (2020-2022), was randomized to 1 of 4 arms that differed by coaching duration (5 months vs 5 months plus 18 months of access to 1:1 coaching) and mode of scientific mentor engagement (structured vs unstructured). Primary outcomes were national-level grant application submissions and awards, and success rate among submitters, assessed via surveys and verified through award databases 30 months after study initiation. ResultsAmong 271 faculty and 96 postdoctoral fellows, 69% submitted at least 1 national application and 32% were awarded at least 1 grant. Submission rates did not differ by experimental arm. Among faculty, the structured engagement of a mentor in coaching was associated with significantly higher odds of any award (adjusted OR = 1.87) and any NIH mentored career development (K) award (adjusted OR = 3.32), and with higher success rates for these proposal types. Extended coaching duration showed a positive but not statistically significant association with faculty awards after adjustment. Neither factor significantly affected outcomes for postdoctoral fellows. Faculty success rates exceeded national NIH-reported success rates for research project grants equivalent to the R01 mechanism (43% vs 17.6%) and K awards (51% vs 36.5%). ConclusionParticipation in a national, cross-institutional, group coaching model yielded high grant submission outcomes, and award rates were higher relative to NIH benchmarks. Structured integration of scientific mentors with the coaching intervention had a greater effect than extended coaching duration on faculty outcomes.

Effective mentorship is essential for strengthening the STEMM (Science, Technology, Engineering, Mathematics, and Medicine) workforce, yet empirical evidence on how mentorship networks are structured and linked to career success remains limited. Here, we analyze mentorship networks among recipients of NIH career development (K) awards to characterize network size, mentor roles, and their associations with mentee-reported outcomes, including potential variation by sociodemographic characteristics. We found that K-awardees rely on mentors beyond their primary advisor, who play varying roles beyond being a Research mentor. Different mentor roles led to different types of mentoring outcomes; while Research mentors were associated with research-related outcomes such as Publications and Grants, career- and psychosocial-related mentoring outcomes were more likely to come from other types of mentors, such as Coaches, Connectors, and Sponsors. Larger networks, as well as having Peer and Identity mentors are additively beneficial for researchers who identify as underrepresented in science more than their counterparts. This study provides large-scale evidence on how mentorship network configurations relate to early-career grant success.

Abstract Objective: Physician-scientists are a diminishing subset of the pediatrician work force, uniquely trained to advance child health through scientific discovery. The current financial landscape of academic medical centers makes traditional resource-intensive training programs untenable. Continued renewal of this critical pediatric workforce requires new resource-efficient methodologies. Thus, we evaluated the impact of the NICHD supported Vanderbilt K12 program, a personalized, intensive training program. Study Design: We tracked academic outcomes for scholars with K12 appointments between January 2015 and December 2022. Our primary outcome was receipt of a mentored career development award (K08 or K23). Successful transition from mentored to independent NIH funding (K to R transition) was used as a secondary outcome. Academic outcomes for faculty participating in a large, institutional training program, the Vanderbilt Faculty Research Scholars program (VFRS), served as a comparison cohort. A complementary qualitative evaluation of the K12 program was performed. Results: K12 scholars earned career development awards (73%) at an equivalent rate to VFRS faculty (71%) and transitioned K to R funding at equivalent rates, 45% of K12 scholars compared to 50% for VFRS. Qualitative analysis demonstrated K12 scholars valued a personalized training program that incorporated self-efficacy and adaptability, key tenets of Social Cognitive Career Theory. Conclusion: Personalized physician-scientist training programs may enhance efficient utilization of research resources in Pediatric academic departments and serve as a viable alternative to traditional programs. Our qualitative evaluation suggests scholar self-efficacy and adaptability can be achieved through a personalized training program.

Background: The rapidly evolving United States (U.S.) health and science policy landscape in 2025 may have heterogeneous effects on academic medicine, biomedical research, physician workforce development and patient care. Research Question: How do academic pulmonary and critical care medicine (PCCM) physicians perceive U.S. health and science policy changes and which changes do they consider most influential on research, trainee education and clinical care? Study Design and Methods: We conducted a prospective survey from March 27, 2026 through April 29, 2026 among selected faculty at U.S. academic PCCM programs as part of the Alliance of Scholars and Professionals for Innovation, Research, and Education in Pulmonary and Critical Care Medicine (ASPIRE-PCCM). Results: Ninety-four of 99 ASPIRE-PCCM panel members completed the survey (response rate 95%), representing 18 of 19 ASPIRE-PCCM wave 1 institutions. All respondents reported the rate of changes in U.S. federal health and science policy in the first half of 2025 as 6 or higher on a scale of 1 ("typical pace of change") to 10 ("most change in my lifetime"), with 89 (96%) evaluating the changes as harmful or very harmful. Respondents identified cancellation of research grants, disruptions and changes to NIH scientific review processes, and changes to visa and immigration policies as key factors affecting early career recruitment. Reported negative effects on quality of care included increased vaccine hesitancy, changing reimbursement structures, and diminished research capacity. State-level and non-governmental efforts, including professional society advocacy, institutional bridge funding and personal advocacy efforts were identified as partially mitigating harms. Despite these concerns, 72% of respondents reported that would still pursue careers in PCCM. Interpretation: Academic pulmonary and critical care physicians reported challenges following recent U.S. health and science policy changes that may be mitigated, but not neutralized, by institutional, professional-society, and individual actions.

Adipose tissue (body fat) is increasingly recognized as a dynamic organ system essential to human health, with distinct depots that serve specialized biological functions. However, this understanding is not reflected in traditional human anatomy curricula, where adipose tissue is typically introduced only as connective tissue and treated in dissection as an obstacle to be removed rather than a structure worthy of study. Anatomy dissection courses have been identified as a source of negative weight bias in medical students, with students reporting disgust toward adipose tissue and frustration with its removal to visualize course-required anatomical structures. Here, we hypothesize that reframing adipose tissue as a functional organ system within anatomy curricula may improve students understanding of human anatomy and mitigate these negative perceptions. To test this, we created a human adipose atlas (AdipoAtlas) by identifying and photographing macroscopic adipose depots in an anatomical donor and organizing these images into an anatomical reference with evidence-based functional descriptions. The atlas was integrated into a human anatomy dissection course, followed by a survey assessing students perceptions of adipose tissue and attitudes related to weight bias, with a concurrent non-dissection anatomy course serving as a control. Students in the adipose-inclusive course reported more positive perceptions and improved understanding of adipose tissue as a multifunctional organ system, while responses related to weight-based discrimination and broader attitudes toward body size were similar between groups. These results support our hypothesis that integrating adipose tissue into anatomy education improves anatomical understanding and may reduce negative weight-related perceptions.

Purpose: Age is the leading factor for age-related macular degeneration (AMD), in which retinal pigmented epithelium (RPE) atrophy is a hallmark. The nuclear factor-erythroid 2-related factor 2 (Nrf2), a major anti-stress transcription factor, decreases in AMD, suggesting bioenergetic stress interacts with age during AMD pathogenesis. We profiled visual function and gene expression of the retina and RPE-choroid with Nrf2 deletion and aging. Methods: We tested 3mo and 14mo Nrf2 null (Nrf2 KO) and Nrf2 WT mice for optokinetic tracking (OKT) and dark-adapted electroretinography (ERG) responses. Eyes were harvested for retina outer nuclear layer (ONL) thickness analysis and retina and RPE-choroid cup RNA isolation for RNAseq profiling. Results: Visual acuity was reduced with aging with lower values in the Nrf2 KO vs Nrf2 WT comparison at 14mo. We observed aged-related reduction of a-, b-, and c-wave ERG amplitudes in both WT and KO mice. Age-related c-wave decline was exacerbated by Nrf2 loss. By RNAseq, two-way ANOVA of RPE-choroid samples revealed differentially expressed genes (DEGs) with age and genotype main effects, as well as DEGs with age and genotype interaction effects. Age DEGs were over-represented in pathways related to inflammation and phagocytosis. Genotype DEGs were associated with alterations in antioxidant and inflammatory processes. Retina RNAseq showed age and genotype DEGs. Conclusions: Ablation of Nrf2 exacerbates age-related declines suggesting that decrease in Nrf2 with aging has functional consequences for inflammation/resiliency. We provide retina and RPE-choroid bulk-RNAseq datasets from the same cohort of mice that can be interrogated to better understand degeneration of retina/RPE-choroid.

Importance: Physician-scientist careers depend on sustained research funding, protected time, and institutional resources. However, data on career-retention attitudes across career stages are limited. Objective: To characterize career intentions and associated research-environment factors among MD-PhD students and early-career physician-scientists. Design: Two independent, national cross-sectional surveys were conducted during nonoverlapping periods in 2025; MD-PhD applicant and matriculant trends were also examined. Setting: Medical schools and academic health centers in the United States. Participants: MD-PhD students and early-career physician-scientists (ECPS). Exposures: MD-PhD exposures included perceived institutional support, training-related mental health impact, and research disruption. ECPS exposures included institutional support, adverse funding-related wellbeing, and direct National Institutes of Health funding-process disruption. Main Outcome and Measures: Primary outcomes were consideration of leaving the physician-scientist pathway among MD-PhD students and consideration of leaving academic medicine or research among ECPS. Secondary outcomes included likelihood of staying in academic medicine, research-intensive career preference, and consideration for leaving the US to continue research. Results: Among MD-PhD programs confirmed to have distributed the survey, 838 student responses were received (estimated response rate of 19.3%); 175 ECPS responses were received, with no calculable individual-level response rate. Among MD-PhD students, 423 of 835 (50.7%) reported considering leaving the physician-scientist pathway; 712 of 837 (85.1%) reported a high likelihood of remaining in academic medicine, and 664 of 836 (79.4%) preferred a research-intensive career. Adequate institutional support was associated with lower odds of considering leaving (adjusted odds ratio [aOR], 0.28; 95% CI, 0.14-0.54) and higher odds of staying in academic medicine (aOR, 5.65; 95% CI, 2.91-10.98). Training-related mental health impact (aOR, 2.34; 95% CI, 1.72-3.19) and research disruption (aOR, 1.83; 95% CI, 1.35-2.48) were associated with greater odds of considering leaving. Among ECPS, 57.7% considered leaving, 52.6% reported a high likelihood of staying, and 42.9% considered leaving the United States to continue research. Adverse funding-related well-being was associated with considering leaving academic medicine (aOR, 3.13; 95% CI, 1.52-6.63) and leaving the United States to continue research (aOR, 5.29; 95% CI, 2.35-13.12). Conclusions and Relevance: Consideration of leaving was common despite continued interest in academic and research-intensive careers. Institutional support, adverse well-being, and research disruptions were associated with career intentions.

Objective: Preclinical studies suggest that irradiation combined with anti-PD-1 antibody (iRT) exacerbates cardiac injury in mice, partly through CD8 T-cell-mediated mechanisms. We investigated radiation-induced PD-L1 expression in cardiomyocytes and explored its potential role in iRT-associated cardiac injury. Methods: AC16 cells were exposed to different doses of irradiation. PD-L1 expression was assessed at different time points by flow cytometry, qRT-PCR, western blotting, and immunofluorescence. Cell viability, apoptosis, and cytokine levels were evaluated. PD-L1 was knocked down using siRNA to assess its function. PBMCs or CD8 T cells were co-cultured with irradiated or non-irradiated AC16 cells at different E:T ratios, with or without PD-1 blockade. A C57BL/6J mouse model of iRT-induced myocardial injury was established, and myocardial PD-L1 expression and T-cell infiltration were evaluated. Results: Irradiation upregulated PD-L1 expression in AC16 cells, with increased membrane and cytoplasmic signals at 48 h and prominent nuclear-associated fluorescence at 72 h. Irradiation increased IL-6, MCP-1, CCL5, and CXCL10 release, decreased IL-10, reduced cell viability, and increased apoptosis. PD-L1 knockdown further increased inflammatory cytokine and chemokine release but did not significantly alter apoptosis or further reduce viability in irradiated AC16 cells. In the presence of a high density of activated CD8 T cells, disruption of PD-1/PD-L1 signaling was associated with a further reduction in AC16 cell viability. In vivo, myocardial PD-L1 expression was increased, whereas CD8 T-cell infiltration remained limited and focal. Conclusion: PD-L1 upregulation in cardiomyocytes may exert an adaptive protective and immunoregulatory role during iRT-associated injury, potentially by limiting inflammatory mediator release and modulating susceptibility to activated CD8 T-cell mediated injury. CD8 T-cell density and activation are the key issues.

Purpose: The neural retina and retinal pigment epithelium/choroid (RPE/Cho) are metabolically coupled. Metabolic dysfunction of the RPE/Cho contributes to photoreceptor death in age-related macular degeneration (AMD). This study aimed to determine how aging, a major risk factor for AMD, alters the proteome and metabolic pathways in the retina and RPE/Cho. Methods: We performed quantitative proteomics on neural retina and RPE/Cho from young (8-week) and old (73-week) C57BL/6J mice. The differentially expressed proteins between old and young, as well as between the retina and RPE/Cho, were visualized using volcano plots and heatmaps, and functional analyses were assessed by Gene Ontology enrichment tools. Results: Aging altered the expression of 85 retinal proteins and 272 RPE/Cho, with 16 shared between tissues. In the aged retina, proteins associated with NADP(H) metabolism, neurodegeneration, and immunity were mostly upregulated, while proteins in lipid metabolism, transcription, and epigenetic regulation decreased. In the aged RPE/Cho, all 21 significantly altered lysosomal enzymes and transport proteins increased, together with complement components, and photoreceptor outer segment proteins. These changes accompanied increases in proteins associated with lipid metabolism and protection against lipid peroxidation, while several glutathione metabolism enzymes decreased. Seven shared proteins involved in lipid and NADP(H) metabolism, changed in opposite directions between retina and RPE/Cho. Comparison of the retinal and RPE/Cho proteomes identified 3073 differentially abundant proteins, of which 2452 maintained tissue-specific differences across both age groups, while approximately 17% showed age-dependent shifts in tissue enrichment. These shifts involved proteins associated with metabolism, extracellular matrix, visual function, lysosomal pathways, and innate immunity. Within the metabolic proteome, several glycolytic and fatty acid oxidation proteins showed greater relative enrichment in RPE/Cho with age, whereas glucose transporters and mitochondrial complexes shifted toward the retina. Conclusions: Overall, these findings show that aging differentially remodels the retina and RPE/Cho proteome, accompanied by altered metabolic specialization between the two tissues.

Background. Cancer cachexia is a multifactorial syndrome characterized by substantial metabolic disturbances including systemic inflammation, and muscle atrophy, which may affect the regulation of iron metabolism in patients. This study aimed to characterize this regulation in two murine models of colorectal and pancreatic cancer, both associated with a high prevalence of cachexia. Methods. Histological, biochemical and molecular analyses were performed in serum, skeletal muscle, liver, spleen and tumor from UN-KC-6141 pancreatic tumor bearing mice and from ApcMin/+ mice (colorectal cancer model), both models exhibiting variable cachexia severity. Results. Both models induce cachexia, but diverge markedly in their regulation of iron metabolism. UNKC mice display reduced plasma iron availability (-9.6% transferrin saturation, p<0.001) without concomitant anemia, while hepatic hepcidin synthesis is upregulated (+100.6%, p<0.001) by inflammation, promoting ferroportin (FPN) downregulation (-42.4%, p<0.001) and iron sequestration in the spleen (+134% total iron content, p<0.001). In the atrophied gastrocnemius, iron concentration is preserved through increased ferritin (+86.6%, p=0.010) and hemoproteins (i.e., myoglobin, CYSC, COXIV) expression. Conversely, ApcMin/+ mice exhibit anemia together with hepatic and splenic iron depletion (-48.4%, p<0.001 and -84.9%, p<0.001, respectively). Hepcidin synthesis is downregulated (-99.9%, p<0.001) despite persistent inflammation, indicating that hypoxia and/or iron-deficiency signals probably predominate over the inflammatory pathway. FPN is markedly decreased (-94.2%, p<0.001) in the iron-depleted spleen despite low hepcidin. In the atrophied quadriceps of ApcMin/+ mice, iron deficiency is associated with decreased FPN and ferritin (-87.9%, p<0.001, and -38.4%, p=0.016, respectively), and increased TfR1 expression (+305%, p=0.107). Finally, tumor iron import may play a significant role in cachexia progression, as suggested by the association between tumor weight, tumor TfR1 protein levels and body weight loss in UNKC mice. Conclusion. These findings reveal cancer type-specific signatures of iron regulation in cachexia, which appear to be more driven by cancer type specific features than by cachexia severity itself, with skeletal muscle iron metabolism impaired mainly when cachexia is associated with anemia. This suggests that the clinical management of iron metabolism in cachectic patients should be guided by the underlying cancer type.

Retinal development is orchestrated by a network of transcription factors that guide multipotent retinal progenitor cells (RPCs) to fate-committed lineages, ultimately producing seven major retinal cell classes. Among these, retinal ganglion cells (RGCs) serve as the sole output neurons of the retina, relaying visual and non-visual information to the brain. RGC specification requires a cascade of transcriptional regulators, including the basic helix-loop-helix (bHLH) factor Atoh7, which confers competence to RPCs, and Pou4f2 and Isl1, which drive terminal differentiation and subtype diversification. Previous studies demonstrate that the SoxC group transcription factors (Sox4, Sox11, and Sox12) are also involved in RGC genesis, but their precise integration into the Atoh7-driven regulatory hierarchy remains undefined. To address this question, we used the retina-specific Vsx2-Cre line to generate Sox4/Sox11 double conditional knockout (dcKO) and Sox4/Sox11/Atoh7 triple knockout (tKO) mice. Immunohistochemistry revealed profound lineage disruption and reduced progenitor proliferation and survival in both dcKO and tKO retinas; not only RGC genesis but also that of horizontal and amacrine (H&As) cells were severely compromised, whereas photoreceptor cells (PHCs) production increased. These results indicate that Sox4 and Sox11 are involved in the coordinated generation of the different early retinal lineages. Like the Atoh7-null retina, RGC precursors still formed in the SoxC dcKO retina, but their genesis was almost completely abolished in the tKO retina. Our findings indicate that the SoxC factors act in parallel with Atoh7 as a major upstream regulatory input to initiate RGC fate. Bulk RNA-seq revealed the SoxC-dependent transcriptional programs and signaling pathways and confirmed the lineage changes demonstrated by marker analysis. CUT&Tag analysis identified the genome-wide binding sites and thereby the target genes of Sox11, further illuminating the mechanisms underlying functions of the SoxC factors in multiple retinal cell states/types during development.

BackgroundInflammatory bowel diseases are complex multifactorial and polygenic conditions with incompletely understood etiology. Transcriptomic and metabolomic studies reveal hundreds of genes and metabolites associated with the inflammatory dowel diseases (IBD), while about 240 genetic loci are associated with these diseases. IBD are accompanied by numerous epithelial lesions including epithelial cells damage and barrier dysfunction. Recent studies suggest that cytoskeleton dysregulation underlies these aspects of IBD. We have previously shown that the disruption of cortical actin filaments and microvilli degeneration are characteristic of Muc2 mouse IBD model. MethodsWe used transmission electron microscopy to evaluate ultrastructural defects in the intestinal epithelium of IBD patients. A probabilistic graphical model (Bayesian network) was employed in order to identify the key gene network hierarchy that might define cytoskeletal architecture among other regulatory events in IBD. The Bayesian network was based on preselected key node genes from open access transcriptomic data obtained in patients with Crohns disease. ResultsElectron microscopy of IBD patients demonstrated disruption of the brush border ultrastructure, similar to the microvillar defects previously described in Muc2 colitis model mice. We highlighted a list of genes contributing to actin polymerization and bundling, of which PREX1 were further confirmed using the transcriptome data obtained from Muc2 mice. ConclusionOur study underscores the utility of Bayesian network analysis in complex cellular phenotypes that helped to identify potential target genes responsible for cortical cytoskeleton and microvilli disruption upon chronic intestinal inflammation.

Objective: One of the strongest genetic associations with coronary artery disease (CAD) risk maps to the metalloproteinase 'a disintegrin and metalloproteinase with thrombospondin motifs 7' (ADAMTS-7) locus. The protein was shown to promote plaque formation and instability. We aimed to generate and evaluate an antibody-based strategy targeting ADAMTS-7 therapeutically to reduce atherosclerotic plaque formation. Approach and Results: A truncated form of human ADAMTS-7 was produced in Nicotiana benthamiana and used as antigen for antibody generation by hybridoma technology. Eight monoclonal antibodies (mAbs) were screened, among which ADAMTS-7-mAb32 (mAb32) demonstrated the highest affinity, as confirmed by surface plasmon resonance analyses and immunoblotting against full-length ADAMTS-7. In vitro, mAb32 inhibited interactions of ADAMTS-7 with its substrates TIMP-1 and SVEP1 in a dose- and time-dependent manner, as assessed by time-resolved Forster resonance energy transfer assays. To assess therapeutic efficacy in vivo, Apoe-/- mice were fed a Western diet for ten weeks and treated with weekly injections of mAb32 or control IgG over the last six weeks. En face aortic Oil Red O staining revealed significantly reduced plaque area in the treatment group, without changes in plasma cholesterol levels or body weight. No evidence of liver or kidney toxicity was observed. Conclusion: Monoclonal antibody-based inhibition of ADAMTS-7 reduced atherosclerotic burden in vivo without affecting lipid metabolism, supporting ADAMTS-7 as a viable therapeutic target in CAD. Further development of mAb32 may provide a cholesterol-independent treatment strategy for atherosclerosis.

Biophysics is a rapidly advancing field with an incredible breadth of topics. Thus, undergraduate biophysics instructors have to strategize and decide what topics they will cover in their courses. Educational institutions utilize a variety of biophysics textbooks. A common deficiency of each of the existing texts is that it serves well a given set of topics (theory, illustrations, practice problems) and leaves out other areas. A typical example includes good theory and problems for thermodynamics and kinetics while presenting molecular dynamics and various spectroscopic methods in a lacking or outdated way. The authors of this manuscript teach a capstone Biophysical Chemistry three-quarter series (Western Washington University/WWU, Bellingham, WA) which ideally should resonate with the general and major-specific courses the students take within their major at WWU. To achieve this goal and to enrich the traditional lecture-based delivery, the instructors have developed and brought together key pedagogical elements: purpose-built online textbook with a uniform structure of the academic content and practice problems, a study sample (oligopeptide) of biophysical significance with a growing set of experimental and computational data and student-centric in-class activities including computer labs. Our Biophysical series emphasizes concepts and methods of computational structural biology (Molecular Dynamics) and spectroscopic approaches (IR, UV and NMR). Here we describe the details of our integrative approach, summarize key outcomes and chart ways to advance the biophysical chemistry series further. Our textbook can be found through LibreText.

IntroductionObesity is a major health issue associated with metabolic and psychological comorbidities, as well as an increased prevalence of disorders of gut-brain interaction (DGBI). Obesity and DGBI share common mechanisms such as inflammation, gut barrier dysfunction, and alterations of gut microbiota, which are all known to be regulated by stress. Glutamine (Gln), which is essential to maintain intestinal integrity and immune response, may counteract these alterations. This study aimed to evaluate the effects of oral Gln supplementation on stress-induced response in obese mice. MethodsSeven-week-old male leptin-deficient ob/ob mice were assigned to four groups: control, chronic restraint stress (CRS), Gln-supplemented, or both CRS and Gln-supplemented. Gln was administered in drinking water for two weeks, and CRS was performed during the final 4 days. Metabolic parameters, intestinal permeability, inflammatory markers, gene and protein expression, and gut microbiota composition were assessed. ResultsStress increased plasma corticosterone levels but had a limited effect on metabolic parameters. In obese mice without stress, Gln supplementation reduced body weight gain, improved body composition and reduced inflammation in the visceral adipose tissue. These effects were lost under stress conditions, with an increase in fasting glycaemia. Stress reduced occludin protein levels, while Gln exerted context-dependent effects, decreasing gene expression of Tjp3, Cldn15 and Ccl2 in unstressed mice but increasing gene expression of multiple tight junction (Tjp2, Tjp3, Cldn12, Cgn, F11r, Marveld2) and inflammatory markers (Tlr2, Myd88, Irf3) under stress. Interestingly, in unstressed obese mice, Gln altered the composition of the gut microbiota, with changes in key bacterial taxa (Thermodesulfobacteriota and Clostridiaceae). This was associated with decreased levels of cecal short-chain fatty acids and increased levels of branched-chain fatty acids. ConclusionIn conclusion, Gln improves metabolic and adipose inflammatory parameters in genetically obese mice. However, these benefits are no longer observed when mice are under stress conditions. Since, Gln has been found to increase fasting glycaemia and colonic inflammation, in association with alterations of gut microbiota.

It is clear that the human retina and its underlying retinal pigment epithelium and choroid (RPE/choroid) form an interdependent metabolic ecosystem, but how metabolism differs between the cone-rich macula and rod-rich periphery remains unclear. Using targeted metabolomics, we quantified 133 metabolites in paired macular and peripheral neural retina and RPE/choroid explants from human donor eyes following short-term culture to restore metabolic activity. Distinct metabolic differences were identified between retinal regions and between tissues. Compared with the peripheral retina, the macula showed metabolic features consistent with greater glycolytic activity, increased NADH availability and higher levels of the neurotransmitter-associated metabolites N-acetyl-aspartate (NAA) and N-acetyl-aspartyl-glutamate (NAAG), consistent with increased energetic and neuronal activity. Compared with peripheral RPE/choroid, the macular RPE/choroid had higher levels of the flavin cofactor FAD together with NAD-related metabolites, including NAD, NADP and NAAD. Comparisons between the neural retina and RPE/choroid further showed that the neural retina was primarily associated with energy production and neurotransmission, whereas the RPE/choroid was associated with cofactor metabolism, nucleotide salvage and lipid metabolism. These findings are consistent with metabolic coupling between the neural retina and RPE/choroid. The macula has metabolic features consistent with high energetic demand, providing a potential metabolic basis for its selective vulnerability in macular disease.