bluuebunny/crossref_metadata_2025_split huggingface dataset:
Exosomal non-coding RNA exhibit sex-specific profiles in salt sensitive hypertension
Authors: Tarhriz Vahideh , Xia Huijing , Eivazi Mortaza , Lazartigues Eric 
Published: May 2024

Exosomes are secreted by various living cells and can be found in all body fluids, especially in cerebrospinal fluid (CSF) and plasma, thereby providing a source of biomarkers for cardiovascular diseases that can be used for patients’ diagnostic. Using high-throughput sequencing technologies, exosomes have been found to contain different RNA populations particularly non-coding RNA. Recently, many non-coding RNA have been recognized as key regulators in the progression of hypertension. In this study we hypothesized that female and male C57BI/6J mice exhibit different microRNA (miR) profiles after deoxycorticosterone acetate (DOCA)+salt hypertension treatment. CSF (10 μl) and plasma (100 μl) were collected from 3-month-old mice and again after 3 weeks of DOCA-salt treatment before exosomes isolation and processing for high throughput small RNA sequencing (Creative Biolabs). Analysis was performed using R language and RStudio ( https://rstudio.com/ ) as an integrated development environment. Global miRNA analysis of CSF exosomes shows different expression miRNA profiles in DOCA+salt male mice compared with their own baseline values. In addition, we observed a remarkable difference in global miRNA expression profile between DOCA+salt male mice and DOCA+salt females (p-value < 2.2e-16), while no significant difference was detected between DOCA+salt females and their baseline values (p-value = 0.6679). Moreover, we observed that there are three clusters of around 200 miR that have different expression levels in DOCA+salt males. Our analysis identified around 74 upregulated miRNA in DOCA+salt males compared to DOCA+salt females. The maximum level of expression belonged to the muscle-specific miRNA (myomiR) miR-1a-3p, miR-206-3p and miR-26a-5p which were highly accumulated in CSF derived exosomes. These top three upregulated miRNA were associated with 282 validated target genes among which, two pivotal genes, ACE2 and ADAM17, were identified in the PPI network, miRTargetLink, and by KEGG analysis. miR-26a-5p was mostly associated with ACE2 and ADAM17 and the mRNA expression of ADAM17 was associated with reduced miR-206-3p. In conclusion, our bioinformatic analysis identified three miRNA and associated them with two key molecules involved in salt sensitive hypertension. Further analysis is likely to highlight more diagnostic biomarkers and potential candidates for the development of new therapeutic approaches. This work was supported in part by a research grant from the National Institutes of Health (HL163588). 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.
A 2-Year Calorie Restriction Intervention Improves Diet Quality Without Compromising Nutritional Adequacy: The CALERIE Trial
Authors: Racette Susan , Garr Barry Valene , DeGraff Jasmyn , Gilhooly Cheryl , Das Sai Krupa 
Published: May 2024

Background: The Comprehensive Assessment of Long-Term Effects of Reducing Intake of Energy (CALERIE) was the first randomized controlled trial of calorie restriction (CR) on biomarkers of aging and cardiometabolic health in humans without obesity. Objective: To determine diet quality and nutritional adequacy during a 2y calorie restriction intervention among healthy adults without obesity. Design: CALERIE phase 2 was a multi-site, randomized controlled trial of 2y of CR vs. ad libitum (AL) control. CR participants received extensive dietary education and support. Food diaries were completed at baseline and months 6, 12, 18, and 24 and analyzed using NDSR. Diet quality was evaluated using the PANDiet diet quality index. Nutritional adequacy was defined using sex- and age-specific Estimated Average Requirement (EAR) or Adequate Intake (AI) criteria for each nutrient. Results: 220 participants completed baseline assessments and were randomized (145 CR, 75 AL; 70% women, age 38±7 y, BMI 25.1±1.7 kg/m2); 86% completed the 2y study. The level of calorie restriction achieved averaged 11.9±0.7% during the 2y intervention. Diet quality improved in the CR group (BL 74.3±5.1%; CR average 76.3±5.3%, p < 0.0001), but not in the AL group ( p < 0.0002 between groups). Nutritional adequacy improved modestly in CR, but not in AL. More than 90% of CR participants achieved 100% of EAR or AI during CR for protein, carbohydrate, 6 of 12 vitamins (A, B1, B2, B3, B6, B12), and 5 of 9 minerals (copper, iron, phosphorus, selenium, sodium). Nutrients for which <90% of participants achieved adequacy included fiber, omega-3 fatty acids, vitamins B5, B9, C, E, and K, and the minerals calcium, magnesium, potassium, and zinc. Conclusion: A 2y CR intervention designed to promote healthy aging and comprised of comprehensive nutrition counseling promoted improvements in diet quality and maintenance or improvements in nutritional adequacy. This research was supported by grants U01AG020478, U01AG020480, U01AG020487, U01AG022132, U24AG047121, R33AG070455, and R01AG071717 from the National Institute on Aging. 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.
Phrenic Motor Neuron Fractalkine Signaling to Nearby Microglia Regulates Phrenic Long-Term Facilitation
Authors: Marciante Alexandria , Mitchell Gordon 
Published: May 2024

Although microglia are traditionally regarded as innate CNS immune cells, recent studies demonstrate that microglia also play key roles in normal CNS functions. For example, microglia engage in bidirectional communication with other cell types ( e.g., neurons) to regulate diverse processes including neuronal excitability, architecture and synaptic plasticity. Although neuron-microglia interactions regulate hippocampal synaptic plasticity, little is known concerning the role of microglia in regulating respiratory motor plasticity. One well-studied form of respiratory motor plasticity is phrenic long-term facilitation (pLTF), a prolonged increase in phrenic motor output following acute intermittent hypoxia (AIH). Whereas AIH consisting of moderate hypoxic episodes (mAIH) elicits pLTF by a serotonin-dominant, adenosine-constrained mechanism, severe AIH (sAIH) elicits pLTF by an adenosine-dominant, serotonin-constrained mechanism. Neurons interact with microglia via release of fractalkine (CX3CL1), activating responses from nearby microglia, the only CNS cell-type known to express fractalkine receptors (CX3CR1). Cervical spinal fractalkine protein injections elicit phrenic motor facilitation similar to pLTF and trigger glial ATP/adenosine release, consistent with the hypothesis that neuronal fractalkine release during hypoxia elicits extracellular adenosine accumulation, thereby modulating AIH-induced pLTF. Here, we tested the hypothesis that phrenic motor neuron fractalkine protein underlies the adenosine-constraint to mAIH-induced pLTF, and drives sAIH-induced pLTF. In adult male Sprague Dawley rats, siRNA knockdown of phrenic motor neuron fractalkine mRNA and protein was achieved via intrapleural injections of siRNAs targeting fractalkine mRNA for 3 successive days. Fractalkine knock-down: 1) enhanced serotonin-dependent mAIH-induced pLTF (100±7%; n=6; vs controls: 50±5%; n=7; p<0.001), but 2) attenuated adenosine-dependent, sAIH-induced pLTF (23±12%; n=5; vs controls: 88±7%; n=7; p=0.001). These findings suggest that hypoxia triggers dose-dependent phrenic motor neuron fractalkine release, activating CX3CR1 on nearby microglia. Subsequent microglial ATP release and conversion to adenosine activates phrenic motor neuron adenosine 2A receptors which: 1) undermines mAIH-induced (serotonin-dominant) pLTF; or 2) reaches levels suffcient to drive sAIH-induced (adenosine-dominant) pLTF. These findings increase our understanding of how spinal intercellular interactions regulate AIH-induced phrenic motor plasticity. NIH R01HL148030 (GSM), NIH R01HL149800 (GSM), NIH T32HL134621-5 (ABM). 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.
The Effect of Gestational PFOS Exposure on Metabolism in the F344 Rat
Authors: Pina Doriann , Holl Katie , Wagner Valarie , Kwitek Anne 
Published: May 2024

Perfluorooctanesulfonate (or PFOS) is a type of PFOA known as (per-and poly-fluorinated alkylated substance) which is a manufactured compound chemical made of a strong carbon-fluorine and sulfonic acid. Due to its hydrophobic and lipophobic properties, industries use it in food wrappers, fire-fighting foam, sweat-proof clothing, and household cleaning products. PFOS exposure is an endocrine disruptor that may induce Metabolic Syndrome (MetS), an association of disorders that increases the risk of significant health issues like heart disease, diabetes, and stroke. Gestational and lactational exposure to Perfluorooctanesulfonate (PFOS) in F344/Stm rats was hypothesized to induce metabolic dysfunction in both female and male offspring. Pregnant F344 rats were given a control diet or diet containing 15mg/kg or 30mg/kg of PFOS; measures of body weight and body composition through nuclear magnetic resonance (NMR) were performed, and tissue was measured at the euthanasia of the dams. Both female and male offspring exposed during pregnancy and lactation were subject to the same protocol with tissue collection at three ages. On both low and high doses of PFOS, the pregnant females showed a decreased body weight gain compared to the control group; males and females also showed decreased body weight at three weeks of age when weaned from the exposed female. At 16 weeks of age, the same patterns were not revealed. The results from NMR suggested a 21% decrease in fat percentage and a 23% decrease in fat for the high exposure in the dams. The offspring at three weeks old showed a reduction in fat mass, fat mass %, and fat-free % in both high and low-dose groups compared to the control group; on the other hand, at 16 weeks old, this trend is not shown. The results suggest that PFOS had a short-term metabolic effect on the offspring at the younger; however, at the adult age, they seem to recover their initial phenotypes. For future studies, it suggested the focus on underlying mechanisms responsible for the rodent's recovery. ASPIRE Program (R25HL168782) Medical College of Wisconsin. 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.
Muscle Fiber Heterogeneity of Insulin Sensitivity in Syntaxin-4 Enriched Skeletal Muscle
Authors: Hoolachan Joseph , Irimia-Dominguez Jose , Bloom-Saldana Elizabeth , Merz Karla , Zhou Chunxue , Bhattacharya Supriyo , Wang Jinhui 
Published: May 2024

Skeletal muscle insulin resistance (IR) is a primary cause for prediabetes, an intermediary condition of higher-than-normal blood glucose levels that left untreated progresses into type 2 diabetes (T2D). A number of contributing factors associated with skeletal muscle IR include aberrant mitochondrial function, elevated reactive oxygen species, inflammation, and reduction in oxidative slow-twitch type 1 skeletal muscle fibers. One leading candidate for reversing skeletal muscle IR is Syntaxin 4 (STX4), traditionally a cell surface transmembrane SNARE protein for GLUT4 vesicle exocytosis that our lab has recently discovered is also localized in the skeletal muscle outer mitochondrial membrane. Importantly, we previously demonstrated that doxycycline-induced skeletal muscle specific STX4 overexpression in obese, prediabetes male transgenic mice (skmSTX4-iOE) reversed IR and mitochondrial dysfunction associated with diabetes muscle. However, the precise mechanisms of how STX4 restores skeletal muscle insulin sensitivity remains to be determined. To test glucose uptake and insulin sensitivity we used the “gold standard” method of hyper-insulinemic-euglycemic clamp in chow-fed skmSTX4-iOE STX4 enriched mice against non-induced controls, we identified improved glucose clearance was only associated with increased insulin sensitivity in type 1 fiber rich soleus muscle. On the other hand, insulin tolerance test studies in doxycycline-induced skeletal muscle STX4 knockout (skmSTX4-iKO) male mice revealed the opposite phenotype, suggesting that STX4 activity in slow-twitch skeletal muscle may be a contributing factor for the reversal of IR. To determine the heterogeneity of STX4 activity between different skeletal muscle fibers, myogenic precursors and intercalating non-muscular tissue in skeletal muscle STX4 enriched skmSTX4-iOE mice, we aim to use our established single nuclear RNA-Sequencing platform (snRNA-Seq) to identify pathways in distinct fiber types associated with STX4 enrichment. Overall, our work aims to determine the functional requirements of STX4 in slow-and-fast twitch skeletal muscle fiber types and identify novel molecular pathways that may serve as useful targets for the reversal/protection against skeletal muscle IR in prediabetes and T2D. This study was supported by grants from the National Institutes of Health (DK067912, DK1129712 and DK102233) and City of Hope Shared Resources Pilot Grant. 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.
Aldo-keto reductase family 1 member A1 contributes to increase the glycolytic metabolism and to accelerate cellular senescence of bone derived mesenchymal stem cells
Authors: Liou Ying-Ming , CHIANG CHEN HAO , LIN YI-HUI , KAO YU-CUIEH , Chen Chuan-Mu 
Published: May 2024

Skeletal aging characterizes the important event where bone loss is accompanied by accumulation of marrow adipose tissues. This event may be due to dysfunctional differentiation of bone derived mesenchymal stem cells (BMSCs) switched from osteoblasts (OBs) to adipocytes (ADs). Our accomplished work demonstrated that Aldo-keto reductase family 1 member A1 (Akr1A1) might serve as an important control to modify the NAD+-dependent SIRT signaling pathway for regulating the adipo-osteogenic lineage differentiation of BMSCs. In this study, we established a senescence cell model for examining whether increased Akr1A1 expression in BMSCs contributes to preferential lineage differentiation to ADs at the expense of osteogenesis by switching on the glycolytic metabolism in senescent cells with D-galactose induction or natural aging in MSCs. Our result suggests that increased expression of the glycolytic enzyme, Akr1A1 contributes to increase the glycolytic metabolism and to accelerate cellular senescence of MSCs. Such information might be useful to help develop the novel therapeutic strategy for treating age-related bone diseases with “perspective Akr1A1 inhibitors”. Grants supported by NSTC (112-2314-B-005 -004 -, NSC-111-2314-B-005-003-), iEGG and Animal Biotechnology Center from the Feature Areas Research Center Program within the framework of the Higher Education Sprout Project by the Ministry of Education (MOE-112-S-0023-A) in Taiwan, and partially by the grants of CYCH-NCHULS110003, CYCH-NCHULS111003, CYCH-NCHULS112002, TCVGH-NCHU 112-7616 to Y-ML. 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.
Unveiling the Mechanisms of Fluid Secretion in Parotid Gland Ducts: CFTR Dependency, Cellular Contributors, and Signaling Pathways
Authors: Catalan Marcelo , Tribiños Felipe , Kirby Nicolas 
Published: May 2024

Compelling evidence indicates that salivary gland ducts mediate fluid secretion, yet the specific cell types responsible for this process remain unclear. Our study aimed to elucidate the fundamental mechanisms governing fluid secretion within ducts and to determine their significance within the context of parotid gland function.Consistent with previous research in exocrine gland ducts, fluid secretion in parotid gland ducts is enhanced when both calcium and cAMP signaling pathways are simultaneously stimulated. Furthermore, we found that duct fluid secretion depends on the activities of CFTR, NKCC1, and the cAMP-activated kinase PKA.To further investigate the cellular contributors to duct-mediated fluid secretion, we generated a mouse model lacking ionocytes in salivary gland ducts using a diphtheria toxin-based approach. Our findings showed that ionocytes play a key role in the process of duct fluid secretion, as their absence resulted in impaired fluid secretion.Overall, our results underscore the significance of ionocytes in facilitating duct fluid secretion, highlighting the essential interplay between calcium and cAMP signaling pathways to achieve optimal secretion. Moreover, our research identified specific basolateral and apical ion-transporting proteins involved in the fluid secretion process. The pharmacological activation of ionocytes may offer a viable alternative secretory pathway when the function of secretory acinar cells is compromised, as observed in pathological conditions that lead to xerostomia. Fondecyt 1211838. 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.
Skeletal Muscle Mitochondrial Response to Diet Differs by Genotype and Sex in an Alzheimer’s Disease Mouse Model
Authors: Johnson Chelsea , Lysaker Colton , McCoin Colin , Kemna Riley , John Casey , Csikos Drummond Vivien , Thyfault John 
Published: May 2024

Introduction: Skeletal muscle mass is reduced in the early stages of cognitive decline. Apolipoprotein ε4 ( APOE4) is the most prevalent genetic risk factor for Alzheimer’s disease (AD) and is associated with faster motor decline in cognitively healthy individuals. APOE4 also increases the risk of insulin resistance and cardiovascular disease, conditions associated with altered bioenergetic function in muscle. However, no studies have examined the role of APOE4 in modulating muscle physiology and the impact this would have on cognitive function. We sought to determine if APOE4 affects skeletal muscle mitochondrial metabolism in response to a high-fat diet (HFD) and if this is reflected in whole-body metabolism. We also examined the role of sex since APOE4 carriers who are female are more likely to develop AD. Methods: Male and female APOE3 (n=32) and APOE4 (n= 29) targeted-replacement mice on a C57BL/6 background purchased from Taconic were fed a standard chow diet until 4 months old. Mice were then switched to a low-fat diet (LFD, 10% kcal fat) or HFD (45% kcal fat) until sacrifice. Glucose tolerance was tested at 7 months by administering an intraperitoneal glucose injection of 1g/kg lean mass. Blood glucose was measured before and 15, 30, 45, 60, 90 and 120 minutes after glucose injection. Resting respiratory exchange ratio (RER) was determined using the Promethion Indirect Calorimetry system at 8 months to determine the relative contribution of carbohydrates versus lipids to whole body metabolism. Mice were sacrificed at 9 months and carbohydrate-driven oxygen consumption was measured at basal, state 3 (ADP), state 3 + glutamate, state 3S (succinate) and uncoupled respiratory states in mitochondria isolated from quadriceps tissue on the Oroboros Oxygraph-2k system. Results: Mice fed a HFD had greater glucose area under the curve (p<0.0001) and reduced RER (p= <0.001) compared to mice fed a LFD, regardless of sex or genotype. At the level of skeletal muscle, APOE4 female mice had increased state 3 (p=0.027) and state 3 + glutamate oxygen consumption (p=0.037) on a HFD compared to a LFD while no diet effect was observed in APOE4 males or any APOE3 mice. Conclusion: We provide evidence that female APOE4 mice are susceptible to diet-induced increases in carbohydrate-driven mitochondrial oxygen consumption in muscle. Future studies are needed to determine the underlying mechanism and consequences of increased muscle mitochondrial activity, which could help identify treatment targets in female APOE4 carriers. BHTP T32 AG078114, P30 AG072973 (Chelsea Johnson). 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.
MyoD Over-expression Reprograms Elephant Seal Fibroblasts into Muscle Cells
Authors: Lam Emily , Torres-Velarde Julia Maria , Allen Kaitlin , Vázquez Juan Manuel , Crocker Daniel , Vazquez-Medina Jose Pablo 
Published: May 2024

Ex vivo tissue culture allows the study of complex cellular mechanisms that are relevant to physiological responses while overcoming the challenges presented by studying animals that are not tractable. In a primary cell culture system, certain proliferating cells can be functionally reprogrammed into other cell types via overexpression of key genes. Dermal fibroblasts can theoretically be reprogrammed into muscle cells, which are often challenging to obtain from wild organisms but offer a unique system to study metabolic responses, by overexpression of the myogenic transcription factor myoD. We derived primary dermal fibroblasts from Northern elephant seal (NES) skin samples and propagated them in primary culture. NES cells respire, stain positive for fibroblast markers (vimentin and PDGFR), and are amenable to electroporation. We overexpressed GFP-myoD in NES fibroblasts and conducted antibiotic selection and purification by FACS. As expected, expression of myoD was higher in transfected cells than in controls according to qPCR analysis (t-test p< 0.05). Treatment with small molecules (CHIR99021, Forskolin and Repsox) enhanced myoD expression. Furthermore, fibroblasts overexpressing myoD also expressed downstream myogenenic markers (myogenin, myosin heavy chain 1 and myosin heavy chain 8). MyoD-overexpressing cells differentiate into myotubes, become multinucleated and stain positively for myosin heavy chain 1. RNAseq analysis confirmed a transcriptional profile of myoD expressing cells, which is unique and different from the naive dermal elephant seal fibroblasts. Establishing differentiated muscle fibers from other mature cell types could provide a unique platform to conduct mechanistic studies in species where muscle tissue samples cannot be obtained from live animals. The Department of Defense National Defense Science and Engineering Graduate (NDSEG) Fellowship. 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.