bluuebunny/crossref_metadata_2025_split huggingface dataset:
Novel Pilot Study Reveals that Heat Therapy Increases Muscle Mitochondrial Quality Control and Respiratory Efficiency in Healthy Human Subjects
Authors: Von Schulze Alex , Fuller Kelly NZ , McCoin Colin , Fernandez Clara Amat , Thyfault John , Geiger Paige 
Published: June 2021

Immense focus has been placed on novel treatment strategies to prevent or reverse insulin resistance and type 2 diabetes (T2D). We have shown that heat therapy (HT) in rodents protects against high‐fat diet induced skeletal muscle insulin resistance and impaired glucose homeostasis. However, it remains unknown whether this effect can be translated to humans. The purpose of this pilot study was to determine if HT via hot water immersion was well tolerated in humans and altered mitochondrial metabolism in skeletal muscle. Healthy male and female participants (n=5/sex) underwent one bout of HT increasing core temperature to 38.5°C for 1h. Muscle biopsies were taken one week prior and 24h post HT to determine whether HT impacted mitochondrial respiratory capacity and mitochondrial quality control via autophagy. HT was well tolerated in all subjects and resulted in acute increases in heart rate and reductions in systolic and diastolic blood pressure. HT did not significantly alter maximal mitochondrial respiratory capacity at basal, state3, state3S, or uncoupled states with either palmitoyl‐carnitine + palmitoyl‐CoA (palmitate) or pyruvate + glutamate + malate (PGM) as substrates. However, HT tended to improve respiratory efficiency as indicated by a reduced coupling control ratio (basal/state3) for PGM. Of interest, this improved efficiency and maintenance of mitochondrial respiratory capacity occurred despite HT‐induced reductions in mitochondrial content as inferred by citrate synthase activity. We believe this reduction in muscle mitochondrial content occured as a result of mitophagy ‐ the targeted degradation of damaged mitochondria. Our hypothesis is supported by the observed reductions (or degradation) of the autophagy/mitophagy related protein, microtubule‐associated Proteins 1A/1B light chain 3B‐ phosphatidylethanolamine (LC3II). For the first time, our data suggest that HT increases mitochondrial quality control (via enhanced autophagy/mitophagy) and enhances respiratory efficiency in human skeletal muscle. These data provide promising results for further analysis in metabolically impaired obese subjects.Support or Funding InformationNIH P30GM122731; Madison and Lila Self Graduate Fellowship (The University of Kansas)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Associations between Aerobic Capacity, Hepatic Bile Acid Synthesis and Fecal Bile Acid Concentration in Humans
Authors: Maurer Adrianna , Mercer Kelly E. , Adams Sean H. , Thyfault John 
Published: June 2021

High aerobic capacity protects against a wide range of metabolic diseases and associates with changes in liver metabolism including enhanced mitochondrial function, although mechanisms remain unknown. Bile acids are now seen as active players in metabolic regulation. We have previously found that rodents with high aerobic capacity potentially have the ability to divert excess acetyl‐CoA away from de novo lipogenesis and toward bile acid synthesis and fecal excretion. To determine if the fitness‐bile acid association manifests in humans, in a preliminary study we tested the hypothesis that female subjects screened for high aerobic fitness (Hi‐Fit, n=10) (VO2peak>45 ml/kg/min) also have increased hepatic bile acid synthesis and fecal excretion compared to those matched for age and body mass with low aerobic fitness (Lo‐Fit, n=10) (VO2peak<35 ml/kg/min). Diet intake, activity level, stool, and blood were measured at baseline before receiving a 1 week standardized, weight‐maintaining eucaloric diet. After the 1week standardized diet, stool and blood were again measured, and an Oral Glucose Tolerance Test (OGTT) was performed. Hi‐fit women tended to have higher bile acid synthesis via measured serum 7α‐hydroxy‐4‐cholesten‐3‐one (C4), a clinically proven surrogate of bile acid production, both during fasting (p≈0.16) and following the OGTT (p≈0.06). However, no significant differences in circulating or fecal bile acid concentrations were found. Future research should consider using direct tracer measurements of bile acid synthesis and excretion, and the utilization of 24 hr+ fecal bile acid measures to quantify total production and excretion.Support or Funding InformationUSDA‐Agricultural Research Service Project 6026‐51000‐010‐05SThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
FGGY Carbohydrate Kinase Domain Containing (Fggy) is Expressed and Alternatively Spliced in Skeletal Muscle
Authors: Izhar Mahnoor , Smith Anastasia , Waddell David 
Published: June 2021

Skeletal muscle atrophy can result from a range of physiological conditions, including denervation, corticosteroid exposure, immobilization and aging. In order to better characterize the molecular genetic events of atrophy, a previous investigation isolated skeletal muscle from mice following 3 days and 14 days of denervation. The gene expression profile of the denervated muscle tissue was analyzed by microarray and compared to control muscle in order to identify novel atrophy‐induced genes. The microarray revealed that FGGY carbohydrate kinase domain containing (Fggy) is expressed in skeletal muscle and is induced in response to denervation. Bioinformatic analysis of the Fggy gene locus revealed two validated alternative isoforms, which we have termed Fggy‐L and Fggy‐S, that have distinct transcription initiation sites. In order to confirm that Fggy is expressed in muscle cells, the cDNAs of the two validated alternative splice variants were cloned from mouse myoblast cells. Interestingly, a novel alternative spice variant for each of the validated alternative isoforms was also cloned from mouse muscle cells, suggesting that at least four Fggy variants are expressed in skeletal muscle. In order to better characterize the expression profile of the Fggy gene, quantitative RT‐PCR (RT‐qPCR) was performed using RNA isolated from muscle cells and primer pairs that distinguish all four alternative Fggy transcripts. The qPCR data revealed that the Fggy‐L transcripts are more highly expressed in proliferating muscle cells, while the Fggy‐S transcripts are more evenly expressed in muscle cells that are both proliferating and differentiated. Western Blot analysis of protein homogenates isolated from muscle cells confirmed that the Fggy‐L isoform is more highly expressed in proliferating muscle cells, while the Fggy‐S isoform is more ubiquitously expressed. In addition, confocal fluorescent microscopy analysis showed that the Fggy‐L full length and novel proteins are localized throughout the cell, while the Fggy‐S full length and novel gene products produce a punctuate pattern throughout the cytoplasm of muscle cells. Finally, ectopic expression of Fggy in cultured muscle cells resulted in significant inhibition of muscle cell differentiation. The characterization of novel genes that are activated during neurogenic atrophy helps improve our understanding of the molecular and cellular events that lead to muscle atrophy and could eventually lead to new therapeutic targets for the treatment of muscle wasting.Support or Funding InformationThe work was support by University of North Florida Transformational Learning Opportunity grants and a University of North Florida Foundation Board Grant to D.W.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ubiquitin Specific Peptidase 3 (USP3) is Expressed in Skeletal Muscle and is Upregulated in Response to Neurogenic Atrophy
Authors: Cruikshank Aimee , Waddell David 
Published: June 2021

Skeletal muscle atrophy is a physiological condition that is caused by a variety of conditions, including immobilization, denervation, spinal cord injury and aging and results in decreased muscle size and strength. To better characterize the molecular genetic events of neurogenic atrophy, a previous study isolated gastrocnemius muscle from mice following 3 days and 14 days of denervation. The gene expression profile in the denervated muscle tissue was then analyzed by microarray and compared to control muscle tissue to identify novel neurogenic atrophy‐induced genes. The microarray data revealed for the first time that Ubiquitin Specific Peptidase 3 (USP3) is expressed in skeletal muscle and is significantly induced in response to denervation at 3 days post‐denervation but returns to baseline expression levels by 14 days post‐denervation. USP3 is a deubiquitinase that has been shown previously to stabilize p53 by deubiquitination. To confirm that USP3 is expressed in skeletal muscle, we cloned the USP3 cDNA from cultured muscle cells. In addition, quantitative PCR (qPCR) was performed to assess USP3 expression levels in both proliferating and differentiated muscle cells and the results demonstrate that USP3 expression levels are relatively equal in both proliferating myoblasts and in differentiated myotubes. Furthermore, Western blot analysis of protein lysates isolated from proliferating and differentiating muscle cells revealed that USP3 is expressed in proliferating myoblasts, but then decreases modestly as the cells differentiate in culture. The discovery that USP3 is induced in response to neurogenic atrophy helps further our understanding of the molecular genetic events of atrophy and may eventually lead to the identification of new therapeutic targets for the treatment and prevention of muscle wasting.Support or Funding InformationThe work was support by University of North Florida Transformational Learning Opportunity grants and a University of North Florida Foundation Board Grant to D.W.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identification and Characterization of Homocysteine Responsive Endoplasmic Reticulum Resident Ubiquitin‐like Domain Member 2 (Herpud2) in Skeletal Muscle
Authors: Arcaro Alexandra , Hewitt Lillian , Waddell David 
Published: June 2021

Skeletal muscle atrophy is a physiological condition that is caused by disparate conditions, including immobilization, denervation, spinal cord injury and corticosteroids and results in decreased muscle size and strength. To better characterize the molecular genetic events of neurogenic atrophy, a previous study isolated gastrocnemius muscle from mice following 3 days and 14 days of sciatic nerve denervation. The gene expression profile in the denervated muscle tissue was then analyzed by microarray and compared to control muscle tissue to identify novel neurogenic atrophy‐induced genes. The microarray data revealed for the first time that Homocysteine Responsive Endoplasmic Reticulum Resident Ubiquitin‐like Domain Member 2 (Herpud2) is expressed in skeletal muscle and is differentially regulated in response to denervation. Herpud2 is related to Herpud1, which has previously been shown to play a role in endoplasmic reticulum‐associated degradation (ERAD) of unfolded ER proteins via retrotranslocation to the cytoplasm and degradation by ubiquitin‐proteasome system. To confirm that Herpud2 is expressed in muscle cells, we cloned the Herpud2 cDNA from cultured cells and performed quantitative PCR (qPCR) to assess Herpud2 expression levels in both proliferating and differentiated muscle cells and the results demonstrate that Herpud2 expression levels are lower in proliferating myoblasts compared to differentiated myotubes. Furthermore, Western blot analysis confirmed that Herpud2 is expressed in cultured muscle cells at the protein level. The discovery that Herpud2 is differentially expressed in response to neurogenic atrophy helps further our understanding of the molecular genetic events of muscle wasting and may eventually lead to the identification of new therapeutic targets for the treatment and prevention of atrophy.Support or Funding InformationThe work was support by University of North Florida Transformational Learning Opportunity grants and a University of North Florida Foundation Board Grant to D.W.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Increased Protein Intake Prevents Elevations in Sclerostin during Short‐Term Diet‐ and Exercise‐Induced Weight Loss
Authors: Murphy Chaise , Marks‐Nelson Eileen , Koehler Karsten 
Published: June 2021

Caloric‐restriction is necessary for efficacious weight loss, but can lead to unwanted losses of lean mass and bone mineral content. High‐protein diets have been shown to protect lean mass during caloric‐restriction and may have similarly preservative effects on bone either through attenuating increases in bone resorption or elevating bone formation.ObjectivesTo determine whether increasing protein intake, in combination with daily aerobic exercise, attenuates elevations in bone resorption markers and reductions in bone formation markers during short‐term severe caloric‐restriction in healthy, active men. We hypothesized increasing protein intake would attenuate elevations in sclerostin and c‐terminal telopeptide (CTX) while maintaining levels of procollagen type I propeptide (P1NP).MethodsIn this randomized crossover trial, participants underwent three conditions: caloric‐restriction with low protein intake (CRLP; 0.8 g/kg body weight (BW)); caloric‐restriction with high protein intake (CRHP; 1.7 g/kg BW) and energy balance control (CON; 1.7 g/kg BW). Caloric restriction was achieved by reducing energy availability, calculated as the difference between dietary energy intake and exercise energy expenditure, to 15 kcal/kg fat‐free mass (FFM), whereas energy availability was maintained at 40 kcal/kg FFM in CON. Throughout all conditions, participants completed daily supervised exercise sessions on a bicycle ergometer set to expend 15 kcal/kg FFM. Before and after each five‐day condition, body composition was assessed via bioimpedance analysis and markers of bone formation and resorption were assessed from fasting blood samples.ResultsBody weight decreased (p < 0.001) in CRLP [−1.97 ± 0.22 kg] and CRHP [−1.80 ± 0.52 kg], but remained stable in CON. Loss of fat mass was significant in CRHP [−1.14 ± 0.23 kg, p < 0.05] but not in CRLP [−0.57 ± 0.48 kg]. Sclerostin was elevated above baseline levels only in CRLP [19.7 ± 22.5 %, p < 0.05] but not in CRHP [3.2 ± 18.3 %] or CON [3.7 ± 26.1 %]. CTX was reduced in CON [−8.9 ± 9.8 %, p < 0.05] but did not change in CRHP [0.1 ± 5.4 %] or CRLP [3.8 ± 14.7 %]. P1NP did not change significantly in any group.ConclusionsElevating protein intake and performing daily aerobic exercise during five days of severe caloric restriction suppressed the elevation in sclerostin, a bone resorption marker, seen in individuals who performed daily aerobic exercise but did not elevate protein intake. This effect mirrors the protective effect of increased protein intake on lean mass.Support or Funding InformationThis work was supported in part by funding from the USDA National Institute of Food and Agriculture (NEB‐36‐083) and a Layman Seed Award from the University of Nebraska Foundation.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
MiR‐29b levels in Cord Blood from Preterm Infants Are Associated with Fetal Inflammation
Authors: Pavlek Leeann R , Vudatala Sundari , Warnement Catherine M , Heyob Kathryn M , Buhimschi Irina M , Buhimschi Catalin , Rogers Lynette 
Published: June 2021

BackgroundPrevious reports have identified decreases in miR‐29b levels in preterm infants that go on to develop bronchopulmonary dysplasia (BPD). The cause of depressed miR‐29b levels is unknown but may reflect an adverse maternal and/or intrauterine environment that led to preterm birth. This study was designed to investigate relationships between antenatal circumstances related to preterm birth and cord blood levels of miR29b.MethodsWe analyzed 127 cord blood collected from consecutive deliveries (n=114) of liveborn infants born ≤30 weeks gestation (median 27.4 weeks: IQR [25.6–29.0]). Data on maternal co‐morbidities, antecendants to preterm birth (e.g., preeclampsia, fetal growth restriction, placental inflammation) were collected prospectively. miR‐29b levels were measured using RT‐PCR and normalized to expression of SP2. Data were analyzed for correlations by Spearman's correlation coefficient or Generalizing Estimating Equations (GEE) for mixed models controlling for twins.ResultsPreliminary results indicate that miR‐29b levels were correlated to umbilical funisitis (p=0.038). Regression analysis revealed miR‐29b is associated with intrauterine growth retardation (p=0.011) and pre‐term labor (p=0.007).ConclusionsmiR‐29b levels are associated with fetal inflammation and poor fetal growth. Maternal health is also related to fetal inflammation and further analyses of the clinical data is likely to reveal new correlations.Support or Funding InformationNIH NICHD R01HD088033This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Deletion of miR‐92a Results in Glucose Intolerance via Impaired Pancreatic Beta Cell Function
Authors: Islam Md Torikul , Tippetts Trevor S , Gogulamudi Venkateswara R , Holland William L , Summers Scott A , Donato Anthony J , Lesniewski Lisa A 
Published: June 2021

MicroRNAs (miRs) are small non‐coding RNA that regulate numerous physiological and pathological processes by repressing protein translation. MiR‐92a, a member of the miR‐17–92 cluster, is altered during development and in a host of disease states, with pancreatic miR‐92a levels reduced in animal models of diabetes. Here, we tested the hypothesis that miR‐92a is a critical regulator of metabolism that modulates pancreatic beta cell function and glucose homeostasis. Glucose tolerance and insulin sensitivity were assessed by glucose (2 g/kg, ip) and insulin (1 U/kg, ip) tolerance tests, respectively in wildtype (WT: C57Bl/6) and whole body miR‐92a (miR‐92a−/−) knockout mice. Deletion of miR‐92a impaired glucose tolerance (p<0.01), but not insulin sensitivity (p>0.05), compared to WT mice. To explore the potential causes of this glucose intolerance, we assessed both fasted insulin and glucose‐stimulated (1 g/kg, ip, 5 min) insulin secretion (GSIS), and found both to be lower in miR‐92a−/− mice compared to WT mice (both p<0.01). To examine if miR‐92a modulates insulin secretory capacity, we administered 10% L‐arginine (1 g/kg, ip) and a β‐adrenergic agonist (CL 316,243: 1 mg/kg, ip) following a 4–6 hr fast and plasma insulin was assessed at time 0, 5 and 15 min. Fasted plasma insulin was ~70% lower in miR‐92a−/− mice compared to WT mice (p<0.01). In response to L‐arginine, plasma insulin was lower in miR‐92a−/− mice compared to WT mice (1.80 ± 0.52 vs 4.57 ± 0.62, 5 min, p<0.01; 1.23 ± 0.42 vs 6.30 ± 2.66, 15 min, p<0.05). Likewise, plasma insulin was ~3 and ~8‐fold lower at 5‐ and 15‐min after β‐adrenergic stimulation in miR‐92a−/− mice compared to WT mice (both p<0.01). Taken together, these results indicate a reduced insulin secretory capacity in response to glucose, amino acid and lipid stimulation in miR‐92a−/− mice. Subsequent histological analysis demonstrated that both average islet size and extrapolated total islet mass were ~50% less in miR‐92a−/− mice compared to WT mice (both p<0.05). In conclusion, our findings support a critical role for miR‐92a in pancreatic beta cell function and whole body metabolic homeostasis.Support or Funding InformationThis work was supported in part by awards from the National Institute on Aging, R01 AG048366, R01 AG050238, and K02 AG045339, and by Merit Review Award 1I01BX002151 from the United States (U.S.) Department of Veterans Affairs Biomedical Laboratory Research and Development Service.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Altered Non‐Coding RNA‐Histone Acetylation Regulatory Circuit Is Associated With Cognitive Impairment via Gut Dysbiosis in Aging Mice
Authors: Behera Jyotirmaya , Kelly Kimberly E. , Tyagi Neetu 
Published: June 2021

Advancing age is the top risk factor for the development of neurodegenerative disorders such as Alzheimer's Disease (AD). However, the contribution of the aging process to Alzheimer's disease (AD) pathology remain unclear. It is also unclear, whether age‐related disruption of the gut‐barrier integrity or dysbiosis causes impaired hippocampal memory. Allyl sulfide (AS), an organic sulfur compound in garlic, has many health benefits and is known for its detoxifying agents. However, the precise role of allyl sulfide in regulating hippocampal memory function at mechanistic levels in mice still needs to be investigated. Here we show that memory disturbances with altered hippocampal chromatin plasticity in the aging brain of the mouse are associated with gut‐microbiota changes and intragastric administration of allyl sulfide retards age‐related defects. To test the hypothesis, we employed 4‐months age‐matched male wild‐type C57BJ/L6 (WT) mice as control group (Young), 24‐months old age‐matched male WT mice group as aging group (AG) and another 24‐months old age‐matched male WT mice group received daily AS at the dosage of 2mg/kg/b.wt through oral gavage (AG+AS). The results show that Young and AG+AS mice showed higher behavioral proficiency and more robust long‐term and short‐term memory compared to AG mice. Gut microbiota is significantly altered as assessed by the denaturing gradient gel electrophoresis (DGGE) analysis in AG group in compared with Young and AG+AS group. The mechanistic study shows that AG mice display an increased plasma level of circulating non‐coding RNA‐Hotair (lncRNA‐Hotair), as assessed by RT2 lncRNA qPCR Assay. To test whether memory impairment correlates with altered hippocampal chromatin plasticity in a lncRNA‐Hotair dependent manner, we first investigated the hippocampal histone acetylation pattern in the AG group. The data suggest that lncRNA‐Hotair causes a specific deregulation of histone H3 lysine 27 (H3K27) acetylation (ac) at the neuronal derived natriuretic factor (NDNF) promoter of hippocampal tissue of AG group and, therefore, fail to initiate a hippocampal gene transcription associated with memory consolidation. However, administration of AS in the AG group could ameliorate gut dysbiosis and memory functions. In lncRNA‐Hotair‐deficient mice, which are protected from age‐related microbiota changes and memory impairment. Furthermore, restoration of NDNF using recombinant NDNF‐therapy reinstates to the recovery of cognitive abilities. In conclusion, our data suggest that lncRNA‐Hotair upregulation and H3K27ac regulatory activity may represent an early biomarker in the aging mouse brain. In addition, AS could serve as potential anti‐aging therapeutics in treating aging individuals.Support or Funding InformationThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.