bluuebunny/crossref_metadata_2025_split huggingface dataset:
Measuring fecal neurotransmitter concentrations using Dried Fecal Spot (DFS)-based LC-MS/MS bioanalysis
Authors: Horvath Thomas , Thapa Santosh , Yacyshyn Mary Beth , Yacyshyn Bruce , Engevik Melinda , Chace Donald 
Published: May 2024

Background: Guthrie implemented dried blood spot (DBS)-based bioanalytical techniques in the 1960s in order to screen for phenylketonuria (PKU) disease in neonates. Historically, the dominant matrix analyzed using the “Guthrie card” platform has been whole blood, but recent advances in device technology now allow for whole blood-to-plasma sample processing prior to drying and subsequent bioanalysis. This project aimed to use this dried matrix technologies in an entirely new direction by performing dried fecal spot (DFS)-based bioanalysis for analytical microbiology and microbiome applications. Homogenized fecal sample extracts were loaded onto quantitative DBS (qDBS) devices and were allowed to dry overnight, then were shipped at ambient shipping temperature. The DBS spots were processed and subjected to LC-MS/MS-based targeted metabolomics using our Glutamate Cycle method (Glu, Gln, and GABA). Methods & Results: Fecal samples were collected from each of the five human subjects enlisted in the following cohorts: 1) healthy controls; 2) patients with diarrhea; and, 3) Clostridioides diffcile-infected (CDI) patients. A volume of ice-cold methanol:water (1:1, v:v) equivalent to a fecal density of 50 mg/mL was added into individual homogenization tubes that each contained 100 mg of 1.4 mm silica beads. Fecal samples were homogenized using 5 bead-beating cycles (4 m/s for each 20 s pulse). After centrifugation (@500 g), a 20-μL sample supernatant volume was loaded onto qDBS devices that yielded an exact 10-μL DFS spot volume on the embedded cards. Dried DFS spots were removed from the device and homogenized in a 200-μL volume of an IS solution. The Glu, Gln, and GABA concentration of each sample was compared against the concentrations measured in the matched frozen fecal extracts. We were able to successfully detect GABA in both the standard extraction method and in our processed DFS samples. Interestingly, we observed large variability between patient samples and there were no statistically significant differences between the cohorts. Conclusion: Collectively, these data demonstrate the utility of our method for measuring the concentrations of Glu, Gln and GABA contained in our processed DFS-based sample sets. This study was supported by the NIH K01K123195-01 (MAE). The Texas Children’s Hospital Department of Pathology provides salary support to Texas Children’s Microbiome Center-Metabolomics Lab staff, and purchased all of the reagents, the consumables and durable supplies, and the liquid chromatography-tandem mass spectrometry (LC-MS/MS) equipment described. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Looper, an updated closed loop feature detection platform for automated neonate cardio-respiratory measurements and data analysis
Authors: Ward Christopher , Al-Madi Mahmoud , Lusk Savannah , Patel Dipak , Memos Nicoletta , Ray Russell 
Published: May 2024

Facemask pneumotachography is a common, precise experimental approach for measuring respiratory outcomes in neonate rodents (P7-8), particularly in autoresuscitation experiments that inform upon Sudden Infant Death Syndrome where failure of autoresuscitation is hypothesized to be a common endpoint. However, current approaches rely on direct, real-time waveform inspection by an observer and their highly variable reaction times to execute responsive changes in assay gas exposures. Looper, represents a novel and significant improvement on typical observer-based interpretation of live data to initiate challenge and recovery gas applications to uncover nuanced features in neonate cardiorespiratory physiology. Leveraging our previously published Breathe Easy software, we have expanded the analytical capabilities of the software to analyze key features within a typical autoresuscitation cycle. Utilizing the software output, we have further developed machine learning approaches that enable real-time prediction of cardio-respiratory failure based distinct waveform features more than 10min in advance, setting the stage for novel experiments to identify critical interventional windows for enhancing resuscitation. R01HL161142 R01HL130249. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
DerP1, a house dust mite protease, directly activates airway vagal afferents and induces airway hyperreactivity after chronic exposure
Authors: Patil Mayur , lurye Karina , Sargeant Marisa , Hadley Stephen , Taylor-clark Thomas , Akopian Armen 
Published: May 2024

Airway hyperreactivity (AHR) is one of the main symptoms of allergic asthma. Current therapeutics used in the treatment of allergic asthma do not target the AHR. Airway sensory afferents have been shown to be critical in AHR, but the mechanisms remain elusive. We have previously shown that house dust mite (HDM) extract, which contains multiple different proteases, can directly activate these airway sensory afferents in the lung via PAR1 receptors. In this study we identified the most potent protease DerP1 is capable of directly and robustly activating airway sensory vagal afferents. DerP1 caused a significant release of CGRP, a neuropeptide released after nerve terminal activation, from the airways. We observed that DerP1 caused a calcium influx in 65% of TRPV1 positive lung specific neurons. We also observed that in our two-photon ex vivo imaging of the vagal ganglia of PirtCre;R26 ‐ GCaMP6s mice, DerP1 caused a robust activation of approximately 60% of naïve airway-specific C-fibers. Calcium imaging of the lung specific vagal neurons from wild type, TRPA1 KO and TRPV1 KO revealed that the DerP1 mediated activation was absent in TRPV1 KO mice neurons. We then found that mice exposed to DerP1 chronically over a period of 12 days (without an adjuvant) had significant AHR and Th2 mediated airway inflammation. Our data suggests that direct activation of airway vagal C-fiber afferents by DerP1 via TRPV1 chronically leads to airway hyperreactivity and Th2 mediated airway inflammation. Warren Alpert Foundation. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Metformin Blunts Bed Rest-induced Skeletal Muscle Ceramide Accumulation in Older Adults
Authors: M. Yee Elena , M.M.P. de Hart Naomi , Mckenzie Alec , J. Petrocelli Jonathan , Kaput Katie , Wahl Matthew , Supiano Mark 
Published: May 2024

Skeletal muscle insulin resistance is an undesirable outcome during periods of physical inactivity in older adults, which can lead to a variety of metabolic diseases. Insulin sensitive tissue (e.g., liver, and skeletal muscle) derived ceramides are well known to promote insulin resistance. Evidence suggests that metformin can regulate tissue ceramide accumulation. Therefore, we hypothesized that metformin treatment during a short-term period of bed rest in male and female older adults would reduce skeletal muscle ceramide accumulation and prevent insulin resistance. Our study design consisted of older adults (66 ± 4.5 y/o placebo, n=10; 71.7 ± 5.1 y/o metformin, n=10) who were subjected to 5 days of bed rest. Participants were randomly assigned to placebo or metformin treatment groups. Drug or placebo treatments started two weeks prior to bed rest and continued throughout the 5 days of bed rest. Prior to and after bed rest, participants underwent a euglycemic-hyperinsulinemic clamp. Muscle biopsies (from the vastus lateralis) were taken prior to and after each clamp both before and after bed rest. We found that insulin-stimulated muscle ceramides were increased after bed rest but were blunted in older adults who took metformin. In addition, muscle insulin resistance correlated with the increase in ceramides. We conclude that metformin was effective to reduce ceramide accumulation in muscle during a period of bed rest in older adults. R21AG064576 awarded to M.J.D. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Drp1 Controls Complex II Assembly and Skeletal Muscle Metabolism by Sdhaf2 Action on Mitochondria
Authors: Zhou Zhenqi , Ma Alice , Moore Timothy , Tran Peter , Shirihai Orian , Hevener Andrea L 
Published: May 2024

The Dynamin-related GTPase, Drp1 (encoded by Dnm1l) plays a central role in mitochondrial fission and is requisite for numerous cellular processes however its role in muscle metabolism remains unclear. Herein, we show that among human tissues, the highest number of significant gene correlations with DNM1L are in skeletal muscle. Knockdown of Drp1 (Drp1-KD) promoted mitochondrial hyperfusion in the muscle of male mice. Reduced fatty acid oxidation and impaired insulin action along with increased muscle succinate was observed in Drp1-KD muscle. Muscle Drp1-KD reduced Complex II assembly and activity as a consequence of diminished mitochondrial translocation of succinate dehydrogenase assembly factor 2 (Sdhaf2). Restoration of Sdhaf2 normalized Complex II activity, lipid oxidation, and insulin action in Drp1-KD myocytes. Drp1 is critical in maintaining mitochondrial Complex II assembly, lipid oxidation, and glucose homeostasis, suggesting a mechanistic link between mitochondrial morphology and skeletal muscle metabolism, which is clinically relevant in combatting metabolic diseases. P30DK063491, DK125354. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Effect of N-Acetyl-Carnosine on Body Weight and Composition
Authors: Milam Norah , Miranda Edwin , Funai katsuhiko 
Published: May 2024

With a globally aging population, the need for treatments that address age related comorbidities are more relevant than ever. Previous research done in our lab has shown that treatment of mice with the compound N-Acetyl-Carnosine (N-Ac-Carn) could partially prevent protein damage caused by lipid peroxidation, and subsequent muscle atrophy and weakness in a mouse-model of inactivity. N-Ac-Carn is endogenously produced, safe, and affordable making it a potentially promising strategy for age-related functional decline. Our current pre-clinical trial aims to comprehensively determine the effcacy of N-Ac-Carn treatment in preventing age-related functional decline. To this end, 18-month-old mice were treated with either normal water (CON) or water supplemented with 80mM N-Ac-Carn (N-Ac-Carn) for 6 months. Targeted metabolomics verified that this treatment enriched all tissues with N-Ac-Carn. Mice treated with N-Ac-Carn maintained higher body weights longer than CON (Interaction: p=0.022). This effect was driven specifically by higher fat mass over time (p=0.016) in female mice. These changes in the N Acetyl Carnosine mice were not explained by an increase in overall food consumption, which remained similar across both groups. Metabolic cage analysis revealed lower VO2 in N-Ac-Carn mice (Group: p=0.002) although this effect was observed in both male and female mice (Interaction: p=0.878). Despite higher fat mass, IPGTT revealed equivocal glucose tolerance in CON and N-Ac-Carn mice. Therefore, this preserved fat mass with aging is not detrimental to metabolic regulation and may be protective against developing other symptoms of frailty. Future analysis will pursue the mechanisms of preserved adipose tissue mass with N-Ac-Carn treatment and if this phenotype is associated with improved health span. R01AG074535-03. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Cystic fibrosis rescued using a reprogrammed porosome secretory machinery
Authors: Cho Won Jin 
Published: May 2024

Cystic fibrosis (CF) is a genetic disorder resulting in the secretion of highly viscous mucus in the airways, leading to lung infection, respiratory failure, morbidity and mortality. CF is attributed to mutation in the CF Transmembrane Conductance Regulator (CFTR) gene that codes for a chloride transporting channel at the cell plasma membrane. More than 2,000 mutations in the CF gene have been identified; however, the ΔF508 CFTR is the most common, accounting for approximately 70% of all CFTR mutations. Our earlier studies demonstrate the CFTR protein to be among the nearly 30 proteins constituting the porosome secretory machinery at the plasma membrane in human airway epithelial mucous-secreting cells. Our recent studies show that stimulated human airway epithelial cells pre-exposed to CFTR inhibitors result in loss of mucus secretion, suggesting the involvement of CFTR in porosome-mediated mucus secretion. To further test the hypothesis that CFTR is involved in porosome-mediated mucus secretion in the human airways, and to develop a therapeutic approach to overcome this defect in ΔF508 CFTR human bronchial epithelial cells, the current study was undertaken. Mass spectrometry and Western Blot analysis of porosomes isolated from WT-CFTR Human Bronchial Epithelial (HBE) Cells and ΔF508-CFTR CF HBE cells, demonstrate a varying loss or gain of several porosome proteins, including undetectable levels of the Ras GTPase activating like-protein IQGAP1 in the ΔF508-CFTR CF cells. This suggested that mutation in porosome-associated CFTR protein additionally affects other proteins within the porosome secretory machinery, negatively impacting mucus secretion. Therefore, to ameliorate defects in mucus secretion in CF, the reconstitution of functional porosomes obtained from WT-CFTR HBE cells into the plasma membrane of ΔF508-CFTR mutant cells was performed. Results from the study demonstrate that porosome reconstitution rescues mucus secretion approximately two- to four-fold more effectively than the currently available CF drugs including TRIKAFTA. Acknowledgements: Work presented in this article was supported by the Viron Molecular Medicine Institute and Porosome Therapeutics, Inc., Boston, MA. Competing Financial Interest: This work is patent protected by Porosome Therapeutics, Inc. The authors hold shares in the company. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Enhancing Learning Opportunities in Laboratory Science Using Virtual Reality Lessons: A Use Case
Authors: Zubek John , Shine Sarah , Warznie Julia , Medina Jenna 
Published: May 2024

Animal dissections are a common approach to teaching anatomy and physiology, but they are costly, pose safety risks, and raise ethical concerns. Alternatives like computer simulations and synthetic specimens have been explored, but they lack the realism of live dissections. Virtual reality (VR) technology provides a promising solution by immersing learners in a virtual environment that closely mimics real-life dissections and allows for skill development and feedback. VR also removes ethical and emotional concerns associated with dissections and enhances learning opportunities. To explore the effectiveness of VR in anatomy and physiology education, we conducted a detailed survey of 48 students in an undergraduate physiology laboratory course. The survey, administered before and after students performed both a VR frog dissection and an in-person frog dissection, was designed to gauge their perceptions, preferences, and learning experiences with both methods. Results indicate that while no participants considered VR a perfect substitute, a majority (80%) still viewed animal dissections as an essential part of science laboratory courses. Interestingly, 43% of participants expressed a stronger preference for live dissections post-experiment, citing greater excitement and perceived informativeness. In contrast, 25% favored VR or alternative methods, highlighting increased comfort and reduced discomfort. A recurring theme was VR's role as a preparatory or alternative tool for those hesitant towards live dissection. This study underscores VR's potential in addressing traditional dissection methods' limitations and emphasizes the need for further research into VR's effectiveness in anatomy and physiology education. We aim to foster dialogue on the use of alternative technologies in enhancing learning opportunities within laboratory and clinical sciences. This project was funded by the Department of Physiology at Michigan State University. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
Specific deletion of FHA domain containing SLMAP isoform in mice impacts embryonic development without affecting Hippo signaling. AP Dias, T Rehmani, M Salih BS Tuana
Authors: Dias Ana , Rehmani Taha , Salih Maysoon , Tuana Balwant 
Published: May 2024

The tail-anchored membrane protein SLMAP3 has been proposed to negatively regulate Hippo signaling and impact cell proliferation. Proteomics have defined SLMAP in complex with STRIPAK a multiprotein interactome consisting of Germinal center kinases and PP2A phosphatase. Data also suggests that the N-terminal FHA domain of SLMAP3 can bind Hippo kinase (MST 1/2) while it‘s coiled-coil regions can integrate PP2A phosphatase to modulate the downstream phosphorylation of YAP/TAZ for cytoplasmic retention. Nuclear YAP/TAZ serve as coactivators of TEAD mediated transcription of genes involved in cell proliferation and organogenesis. The physiological importance of SLMAP3 and it‘s in vivo function in regulating Hippo and organ development remains to be determined. We report that mice with specific deletion of SLMAP3 isoform were embryonic lethal where most organs including muscle, brain, spinal cord, kidney, lungs, intestines, cartilage and bone exhibited severe developmental deficits. The SLMAP3 deficient embryos were ~30% smaller (p<0.05) with small organs compared to wild types. The analysis of Hippo components MST1/2 and YAP/TAZ indicated no changes in their phosphorylation status in embryos or tissue such as muscle and the neural tube due to the loss of SLMAP3. RNA-Seq of whole embryos showed no significant changes to downstream targets of Hippo signaling in SLMAP3-KO. In addition, SLMAP3 loss had no obvious effect on cell proliferation assessed with Ki-67/pH3 staining in embryogenesis. Mouse primary embryonic fibroblasts (MEFs) isolated from embryos deficient in SLMAP3 had no changes in YAP and MST1/2 phosphorylation or proliferation while RNA-seq analysis indicated no changes in TEAD regulated gene activity. Further, phospho-proteomics of MEF lysates revealed no changes in Hippo signaling in SLMAP3 loss, nor in pathways associated with cell proliferation or death. Similarly, knockout of SLMAP3 in myoblast displayed no changes in Hippo signaling or proliferation capacity. These data indicate that SLMAP3 is critically important for embryonic development through unique mechanisms and do not implicate the involvement of Hippo signaling. Supported by CIHR. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.
DNA methylation by DNMT3a drives endothelial inflammation and death during acute lung injury
Authors: Akhter Md Zahid , BR Vijay Avin , Joshi Jagdish Chandra , Mehta Dolly 
Published: May 2024

Vascular endothelium, the inner most lining of all blood vessels, controls the transport of nutrients, protein, water, and leukocytes across the vessel wall and thereby maintains lung tissue and fluid homeostasis. Damage to endothelial integrity caused by mediagenic, inflammatory, or toxic agents results in pulmonary edema, inflammation, organ failure, and high lethality. A key pathogenic characteristic of vascular injury involves an excessive loss of endothelial cells (EC) accompanied by the generation of inflammatory cytokines. Genome-wide analyses of DNA methylation, the most enduring epigenetic modification of DNA, and its impact on EC transcriptomes and cellular functions, still lack comprehensive understanding. We demonstrate that lipopolysaccharide (LPS) elevated DNA methylation in EC within the lungs, primarily through a mechanism dependent on DNA methyltransferase 3a (DNMT 3a). Mice with endothelial cell-specific deletion of DNMT3a ( EC-DNMT3a−/−) promptly recovered from lung injury, attributed to a reduction of EC death. Furthermore, we observed that the deletion of DNMT3a in EC led to an increase in the expression of the apoptosis inhibitor B-cell lymphoma 2 (BCL2). Collectively, the data implies that inhibiting DNMT3a activity plays a crucial role in reshaping the EC methylome and promoting BCL-2 activity. This, in turn, facilitates the generation of reparative EC and contributes to the resolution of acute lung injury (ALI). These findings offer insights into potential targets for preventing ALI. American Heart Association (AHA) Postdoctoral Fellowship (19POST34450241). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.