bluuebunny/crossref_metadata_2025_split huggingface dataset:
Exercise Mitigates Cognitive Functions through Mitochondrial Remodeling in Type‐2 Diabetes
Authors: Kalani Anuradha , Chaturvedi Pankaj , Winchester Lee J , Tyagi Suresh C , Tyagi Neetu 
Published: November 2023

Abnormal brain mitochondrial functions result in cognitive impairments during type‐2 diabetes (T2D). Exercise has been recommended by clinicians to diabetics as a secondary protective therapy; however, its effect on alleviating brain functions through mitochondrial interventions is not much explored. Therefore, we hypothesize that exercise mitigates T2D‐induced neurodegeneration and cognitive functions through mitochondrial remodeling. To test this hypothesis, we used obese diabetic db/db (leptin receptor knockout) and db/+ mice as controls, with and without exercise. The mice were exercised at a constant speed of 7 meters/min for 5 days a week/total 8 weeks. Diabetic mice with exercise had reduced total body weight (R2=0.9154, F=72.15, p<0.001), blood glucose (R2= 0.9264, F=83.92, p<0.001), liver weight (R2=0.9703, F=217.6, p<0.001) and improved heart weight (R2=0.7572, F=20.79, p<0.001) as compared to other experimental groups. However, no significant differences were observed in the weight of kidney, brain, and lungs. We next assessed cognitive and behavior alterations by open field, novel object recognition, Y‐maze, and hot plate analgesia tests. Our results suggest significant improvement in behavior and cognitive functions after exercising diabetic db/db mice as compared to non‐exercised db/db and control littermates. Results with RT2 profiler PCR array, designed especially for mitochondrial genes, confirmed improvements in mitochondrial transport, dynamics, and regulatory genes in exercised db/db brain as compared to non‐exercised db/db brains. The involvement of mitochondrial remodeling in db/db mice following exercise was further confirmed by significant increase in ATP production, membrane potential, copy number as well as decrease in reactive oxygen species when compared with non‐exercised db/db and control mice. Furthermore, considerable reduction in TUNEL reactivity was determined in intact mitochondria isolated from exercised db/db mice as compared to non‐exercised and control mice. In addition, the effect of exercise on neuronal survival in diabetic brain was confirmed by decrease in fluoro‐jade C reactivity that particularly stains degenerating neurons. Taken together, our results indicate myriad beneficiary effects of exercise over metabolic and mitochondrial pathology in diabetes. Furthermore, our findings suggest regular exercise mitigate neurodegeneration and cognitive disorders thereby improving total mitochondrial health.Support or Funding InformationThis work was supported by NIH grant HL107640‐NT.
Defective autophagy causes a maladaptive cardiac phenotype to exercise that leads to premature death and FGF21‐mediated protection against obesity and insulin resistance
Authors: Lira Vitor A. , Kronemberger Ana , Call Jarrod A. , Caster Hannah M , Pereira Renata O. , Laker Rhianna C. , Zhang Mei , Yan Zhen 
Published: November 2023

Autophagy is induced by exercise in skeletal and cardiac muscle. Skeletal muscle autophagy is required for exercise‐induced metabolic adaptations; yet, impaired skeletal muscle autophagy may cause a compensatory ER stress response that protects against diet‐induced obesity and insulin resistance. The functional importance of cardiac autophagy to the benefits of exercise is uncertain. Here, we generated heart & muscle‐specific Atg7 knockout (Atg7h&mKO) mice to gain further insight into this apparent paradox related to skeletal muscle autophagy and to define the role of cardiac autophagy in exercise‐induced protection against obesity and insulin resistance. Despite exhibiting minimal autophagy in skeletal muscle and heart, Atg7h&mKO mice presented normal exercise capacity and glucose tolerance with a slight reduction in body weight (~5%) at 12–14 weeks of age. After 12 weeks on high‐fat diet (60% calories from fat) sedentary Atg7h&mKO mice gained weight and developed insulin resistance like control littermates (Atg7fl/fl). Exercise (voluntary wheel running) reduced the degree of diet‐induced obesity and glucose intolerance in Atg7h&mKO mice more efficiently; but surprisingly led to premature death of ~40% of these animals. This reduced survival was likely caused by a maladaptive cardiac phenotype as exercise induced physiological cardiac hypertrophy and prevented diet‐induced fibrosis in the hearts of Atg7fl/fl, whereas it exacerbated fibrosis and caused pathological hypertrophy in the hearts of Atg7h&mKO. Interestingly, this maladaptive cardiac phenotype to exercise was accompanied by improved skeletal muscle insulin sensitivity, increased Glut4 mRNA in adipose tissues, and elevated Pgc‐1a and Ucp1 mRNA in brown adipose tissue of Atg7h&mKO suggesting a potential cross‐talk between the heart and peripheral tissues. Analysis of serum indicated a robust increase in circulating FGF21 levels in Atg7h&mKO that exercised, which were accompanied by marked upregulation of Fgf21 mRNA expression exclusively in the heart. Examination of β‐Klotho mRNA, a co‐receptor required for FGF21 signaling, revealed a ~2‐fold upregulation exclusively in white and brown adipose tissues. Our results indicate that autophagy plays a critical role in exercise‐induced protection against cardiac dysfunction in obesity and diabetes. Impaired cardiac autophagy causes a maladaptive phenotype to exercise that leads to premature death, but also potentiates the peripheral metabolic benefits of exercise, at least in part, via a heart‐adipose tissue signaling axis involving FGF21.Support or Funding InformationObesity Research and Education Initiative (OREI) and the Fraternal Order of Eagles Diabetes Research Center (FOEDRC) at the University of Iowa (V.A.L.) and U.S. National Institutes of Health (NIH) grant AR050429 (Z.Y.)
Air puffs as refinement of electric shocks for stimulation during treadmill exercise test
Authors: Lamy Sébastien , Cartoni Cristina , Bichat Arnaud , Combe Roy , Doenlen Raphael , Warot Xavier 
Published: November 2023

Endurance treadmill test is a convenient method for easily and systematically evaluate exercise performance and capacity in mice. Although mice are naturally prone to run, an aversive stimulation is required to force all animals to stay engaged in the test. This is typically achieved using mild electrical stimulation on a dedicated grid at the rear of the running belt. Another system replacing the electric stimulation by air puff was newly developed. This system seems to be an ethical evolution in regards of the 3R's principles since the electrical aversive stimulus is replaced by a less noxious stimulus. Exhaustive endurance protocol was used to compare the two different treadmill systems. This test consists to expose mice to a long lasting middle intensity exercise protocol until they reach exhaustion. In addition of the exercise performance readouts, the adaptive mechanisms established to supply fuel to skeletal muscle during exercise and stress were respectively assessed by the analysis of biochemical parameters and markers of stress produced by the hypothalamic‐pituitary‐adrenal axis. The results obtained for the exhaustive protocol are comparable in terms of behavioral and physiological parameters (e.g. running distance, running time and physiological decrease of triglycerides and hepatic glycogen level). Interestingly, corticosterone and adrenocorticotropic hormone (ACTH) levels were similar between the two types of stimulation. These results demonstrate that air puff stimulation could be an interesting alternative for the replacement of the electric stimulation during an exhaustion protocol. However, the stress level does not seem to be reduced when using air puff stimulation.Support or Funding InformationEcole Polytechnique Fédérale de Lausanne, School of Life Sciences, Center of PhenoGenomics
3D Virtual Histology of Trabecular and Cortical Architecture in Living Human Vertebrae
Authors: TANIOKA HISAYA , KAGA KIMITAKA 
Published: November 2023

PurposeToprovide a non‐large‐scaled device but a small‐sized device. Less‐expensive and simple device that can be utilized for making 3D trabecular and cortical structure of living human vertebrae, and to learn about the bone strength from trabecular structure obtained by this created method.Subjects and MethodsThis study involved 15 patients who underwent body CT exams for their diagnosis. Scanning of the body was performed using collimation and pitch values of 1.00 mm and 1.00 respectively with bone algorithm. The field of view was 25.0 cm using a 512 matrix. After transferring the image data to a CT work station, 3D visualization based on interactive direct volume rendering was made using the current distributed software. The defined threshold CT values of this image processing were following; maximum value is 450 HU and minimum value is 100 HU.Results3D virtual histological image of the vertebra was shown in Figure. Morphometric parameters as trabecular number (Tb.N.), trabecular separation (Tb.Sp.), and trabecular thickness (Tb.Th.) were frequently used and their measured values were as followed; TbN=1.00 ± 0.10, TbSp=0.57 ± 0.07 mm, and TbTh=0.45 ± 0.03mm. These values showed significant correlations with the properties of the vertebral bone in the multiple studies.ConclusionThis 3D virtual histological method is very simple. We can get topology and morphological parameters of living vertebrae under regular CT exams. The data of morphometric parameters suggest that this method has potential to characterize the structured implications of metabolic bone.Support or Funding InformationCompeting interests: none Sponsorship: none Funding source: none
The activation on serotonin 2B receptor (5‐HT2BR) is essential for osteoclastogenesis
Authors: Huang Yi‐Chieh , Lin Shiuan‐Yea , Lu Tzong‐Shi , Huang Bin , Lin Chih‐Lung , Fu Yaw‐Syan 
Published: November 2023

Osteoporosis is the common diseases in elderly people, especially menopausal women. Excessive osteoclastogenesis is one of the main reasons to cause osteoporosis. Recent clinical researches indicated that the patients’ long‐term use of Selective Serotonin Reuptake Inhibitor (SSRI) as anti‐depression drugs usually with higher risk to suffer osteoporosis, and that indicated serotonin (5‐HT) may involve in the regulation of bone metabolism. Some previous reports showed that different types of 5‐HT receptors are expressed in both osteoblasts and osteoclasts. This study was focus on the effects of 5‐HT2BR in osteoclastogenesis. Using RANKL induced osteoclast formation by RAW 264.7 cells as the model, our data showed a positive correlation between the osteoclastogenesis with 5‐HT and 5‐HT2BR agonist treatment and that decrease SirT1 expression. Conversely, treatment with specific 5‐HT2BR antagonist significantly reduced the osteoclast numbers and increase SirT1 expression, and treatment with SirT1 agonist significantly reduced the osteoclasts. In conclusion, our study suggested that 5‐HT2BR play an essential role in osteoclastogenesis and SirT1 may be involved in regulating osteoporosis. Our observations might also provide another direction of prophylaxis and therapy for osteoporosis.Support or Funding InformationKMU‐O105001, KMU‐TP104E34, KMU‐TP104E13
Heat Shock Protein 72 Regulates Mitochondrial Integrity and Function in the Prevention of Hepatic Insulin Resistance
Authors: Rogers Robert S. , Wheatley Joshua L. , Archer Ashley E. , Thyfault John P. , Geiger Paige C. 
Published: November 2023

Nonalcoholic Fatty Liver Disease (NAFLD) encompasses a spectrum of disorders characterized by an accumulation of lipids in the liver, and is a significant risk factor for the development of insulin resistance and type 2 diabetes. Heat shock proteins (HSPs) can effectively prevent high fat diet‐induced glucose intolerance and skeletal muscle insulin resistance, improve oxidative capacity and increase mitochondrial function. The purpose of the current investigation was to determine whether HSP72, in particular, is protective against the development of hepatic triglyceride accumulation and insulin resistance. Male Wistar rats were fed a high‐fat diet (HFD) and received either weekly heat treatment (41°C for 20 min) or sham treatment (37°C for 20 min). In addition, primary hepatocytes were isolated from male C57BL/6 mice and subjected to either a single heat (42°C for 30 min) or sham (37°C for 30 min) treatment, transfected with HSP72 siRNA or exposed to insulin (60 μU/ml). Confocal microscopy images were obtained from cells incubated with tetramethylrhodamine (TMRE) and MitoTracker Green to assess mitochondria functionality. Weekly in vivo heat treatments reduced liver triglyceride accumulation, increased HSP72 levels, and modulated the protein levels of microtubule‐associated protein light chain 3 (LC3‐II) and p62 – two markers of autophagy. HSP72 siRNA in primary hepatocytes resulted in reduced mitochondrial function, reduced phosphorylation of AMP‐activated protein kinase (AMPK) and Akt, increased Parkin levels, and decreased LC3‐II levels. Our results indicate that weekly heat treatments protect against liver triglyceride accumulation in rats, and data from primary hepatocytes suggest a critical role for HSP72 in maintaining hepatic mitochondrial integrity and function. Our findings identify potential new HSP72‐mediated therapeutic targets for the regulation of hepatic mitophagy and prevention of metabolic disease.Support or Funding InformationThis work was supported by National Institutes of Health Grants AG‐031575, DK‐088940, the University of Kansas Medical Center Biomedical Research Training Program, and core support was provided by National Institute of Child Health and Human Development Grant HD‐002528.
Altered metabolism in RGS2 KO mice
Authors: Lee Katherine , Gros Robert , Chidiac Peter 
Published: November 2023

Regulator of G‐protein signaling 2 (RGS2) is known to inhibit Gq‐ and Gs‐mediated GPCR signalling. We have previously reported that RGS2 knock out mice (rgs2−/−) exhibit a lean phenotype compared to their wildtype (WT) counterparts, with significantly reduced fat deposits, leptin levels, and serum lipids. The objective of this study is to further elucidate the role of RGS2 in metabolism. CLAMS metabolic cages were used to measure various indices in 1 month‐, 3 month‐, and 12 month‐old rgs2−/− and WT mice. Animals were acclimatised in metabolic cages for 24 hours (12‐hour light and dark cycles), followed by another 24 hours of metabolic recordings.At both 3 months and 12 months, rgs2−/− mice displayed increased rates of respiratory intake (vO2) and expiratory output (vCO2), particularly during the dark cycle, suggesting increased metabolic activity in the knockouts. Furthermore, calculated respiratory exchange ratios (RER) suggest an increase in carbohydrate rather than fat or protein metabolism in rgs2−/− mice of all three age groups compared to WT. Hyperphagia, as well as increased water intake, were also observed in the knockouts at 3 and 12 months. Even with significantly increased food and water intake, 3 and 12 month‐old rgs2−/− mice maintained lower body weights compared to WT, and displayed comparable ambulatory activity.In addition, a 30% diet restriction study in which 3 month‐old mice were fed once daily, followed by metabolic cage measurements, was performed to determine possible alterations in metabolism following changes in food intake. Compared to our previous ad libitum studies, the difference in vO2 and vCO2 between WT and rgs2−/− mice was further exacerbated. In addition, RERs were significantly higher in rgs2−/− mice, and allude to the possibility of elevated carbohydrate metabolism in rgs2−/− mice. Ambulatory activity remained comparable to WT. Interestingly, diet restriction appears to have exacerbated the hyperphagic phenotype of rgs2−/−mice – although body weights remained stable, rgs2−/−mice consumed 50% of their daily food amounts at a 4 times higher rate than WT, and displayed hoarding behaviour.The lack of changes in overall activity between genotypes in both ad libitum and diet restricted mice suggests that weight loss in rgs2−/− mice is due to internal energy metabolism rather than overt physical activity. These results suggest that RGS2 may play an important role in the regulation of body metabolism.Support or Funding InformationCanadian Institutes of Health Research, Heart and Stroke Foundation of Ontario
The Effect of JAK‐Inhibitor Tofacitinib on Intestinal Epithelial Barrier Function
Authors: Sayoc Anica , Krishnan Moorthy , McCole Declan F. 
Published: November 2023

Tofacitinib is an FDA‐approved oral Janus kinase (JAK) inhibitor that modulates the JAK‐STAT signaling cascade activated by inflammatory cytokines, and has shown clinical benefit in trials on ulcerative colitis (UC) patients. However, the effect of this drug on intestinal epithelial cells (IEC) and intestinal permeability, which is increased in UC, has not been elucidated. The aim of this study was to determine if tofacitinib can reduce IEC permeability caused by the inflammatory cytokine, IFN‐γMethodsT84 intestinal epithelial cell lines were cultured on transwells for 12–13 days. Preventive effects of tofacitinib were determined by pre‐treatment with tofacitinib (50μM) apically (to mimic oral administration to patients) for 1 hour followed by basolateral administration of IFN‐γ (1000U/ml) for 24 hours. The concentration of tofacitinib was based on a dose response (2μM, 10μM, 50μM) study on transepithelial electrical resistance (TEER) upon IFN‐γ challenge. Therapeutic effects of tofacitinib were studied by treating T84 cells twice with tofacitinib (16.7μM) at 6 and 18 hours post‐IFN‐γ treatment (1000U/ml). Timing and dosage of tofacitinib were chosen from post‐IFN‐γ dose‐response studies. TEER and 4kD FITC‐dextran permeability were measured to indicate changes in paracellular permeability to ion flux and macromolecules, respectively.ResultsTofacitinib prevented IFN‐γ‐induced increases in TEER (p<0.001; n=3) and 4kD FITC‐dextran permeability in IECs and maintained the barrier integrity at levels comparable to untreated and vehicle (DMSO)‐treated controls. Although one dose of tofacitinib 24 hours post‐IFN‐γ had no effect on TEER, administration of tofacitinib at two time points after IFN‐γ partially rescued IECs from the effect of IFN‐γ on TEER and 4kD FITC‐dextran permeability compared to untreated and vehicle‐treated controls. These data indicate that tofacitinib can restrict intestinal epithelial barrier defects induced by an inflammatory cytokine. This may potentially contribute to the beneficial effects of tofacitinib observed in clinical trials in UC patients.Support or Funding InformationSupported by NIH R01 DK091281 (DFM)
Intestinal Epithelial IL‐15 Overexpression Enhances γδ T Cell Mucosal Surveillance
Authors: Edelblum Karen L , Jabri Bana , Turner Jerrold R 
Published: November 2023

Intraepithelial lymphocytes (IEL) expressing the γδ T cell receptor bridge innate and adaptive immunity through their high level of constitutive activation and surveillance‐like migratory behavior. Although epithelial IL‐15 is critical for the development, proliferation and survival of γδ IELs, the role of IL‐15 in γδ IEL migration remains unclear. Transpresentation of membrane bound IL‐15 requires cell‐cell contact between activating and responder cells, thus we tested the hypothesis that compartmentalized IL‐15 overexpression within the intestinal mucosa directly affects γδ IEL localization. To this end, GFP γδ T cell reporter mice were crossed to transgenic (Tg) mice overexpressing murine IL‐15 either in the intestinal epithelium (vil‐IL‐15) or lamina propria via a MHC class‐I restricted promoter (Dd‐IL‐15). Consistent with the role of epithelial IL‐15 in IEL proliferation, vil‐IL‐15 Tg mice exhibited a 61% increase in γδ T cell number compared to wildtype (WT). Time‐lapse video microscopy revealed enhanced γδ IEL migration into the epithelium in vil‐IL‐15 Tg mice relative to WT (44±8.0% vs 32±3.3%) leading to increased γδ IEL/epithelial interaction (4.5±0.2 vs 3.5±0.2 interactions/hr). In contrast, the frequency of γδ IELs within the intraepithelial compartment (25±6.8%) is reduced in Dd‐IL‐15 Tg mice, resulting in increased γδ T cell localization to the lamina propria. Notably, more epithelial apoptosis was observed in Dd‐IL‐15 Tg mice than in WT. These data indicate that epithelial IL‐15 production plays an important role in maintaining both γδ IEL number and epithelial surveillance, whereas sustained γδ T cell recruitment to the lamina propria may adversely affect epithelial survival.Support or Funding InformationFunding: NIH, RBHS Chancellor Scholar Award
Emerging regulatory roles of Wnt1 in host‐pathogen interactions in inflammation and colon cancer
Authors: Liu Xingyin , Chen Hongshan , Zhang Yongguo , Lu Rong , Liu Qisha , Sun Jun 
Published: November 2023

Salmonella infection is a major public health concern and colonization in humans can be chronic as carrier state. The Wnt signaling plays an essential role in gastrointestinal epithelial proliferation. Our previous studies have demonstrated that both Wnt2 and Wnt1 protein secretion in epithelial cells were elevated after bacterial infection (Liu et al., 2011 and 2012). Bacteria modulate the host by injecting bacterial effectors to the host cells. AvrA is a Salmonella protein that is transported into intestinal epithelial cells by a type 3 secretion system. Our own studies, as well as those of other investigators, have demonstrated that AvrA influences eukaryotic cell pathways by altering ubiquitination and acetylation of target proteins to modulate inflammation, epithelial apoptosis and proliferation. Deubiquitination properties of AvrA leads to upregulation of the Wnt/beta‐catenin signaling pathway, which is critical in initiation of colon cancer. In the current study, we report that Wnt1 plays a role in infection and colon cancer. Different from the bacteria‐induced increase of Wnt2 and Wnt11, pathogenic Salmonella colonization significantly reduces the level of Wnt1 in intestinal epithelial cells in vivo and in vitro. We observed that Wnt1 expression level were down‐regulated by Salmonella infection, but stabilized under AvrA‐deficient Salmonella iin the intestine of Salmonella‐colitis mice. In a chronic Salmonella infected cancer model, the wnt1 protein level is decreased in AvrA+ infected group, consistent with our observation in the Salmonella‐colitis animal model. Furthermore, we assess the functional role of down‐regulation of Wnt1 in the inflammatory response and colon cancer progression. We found that Wnt1 is involved in the regulation of inflammatory pathway in Salmonella‐epithelia cell interaction. Moreover, down‐regulation Wnt1 promotes cancer cell invasion and migration. Interestingly, we found the Wnt1 is down‐regulated in colon cancer patients, which is correlated with colon cancer progression. The results suggest Salmonella effector AvrA and Wnt1 may functionally interrelate in the development of colon cancer. Growing interest and accumulating data on human microbiota implicate that host‐microbe interplay has an important role in the development of colon cancer. A recent study reported that antibody against Salmonella flagellin was higher in colorectal cancer and pre‐cancer cases than controls in two distinct populations in US and the Netherlands and that dietary intake is the one of the mediating factors, suggesting a potential link of Salmonella to colorectal cancer. Our study provides novel insights into mechanisms by which the bacteria effector regulates Wnt1 expression and potential contributes to infectious‐associated colon cancer.Support or Funding InformationStarting package of Nanjing Medical University to Xingyin Liu and NIH R01 to Jun Sun