This study aimed to determine whether sodium nitrite supplementation prevented chronic stress‐induced cerebrovascular dysfunction and cognitive decline. We hypothesize that nitrite supplementation will prevent the oxidative environment and cerebrovascular dysfunction associated with chronic stress and maintain cognitive health. Eighteen‐week‐old male/female C57BL/6 mice underwent 8 weeks of control conditions or unpredictable chronic mild stress (UCMS) with or without sodium nitrite (50 mg/L) in the drinking water. Excised middle cerebral arteries (MCA) were mounted in a pressurized myobath and exposed to increasing concentrations of acetylcholine (ACh). Nitrite supplementation prevented the UCMS‐induced impaired ACh response in the MCA. We examined xanthine oxidoreductase (XOR) as a potential mechanism by determining XOR protein abundance, activity, and hydrogen peroxide production in the liver and brain. Nitrite supplementation prevented the development of an oxidative environment within the liver, brain and cerebrovasculature. Assessment of working memory revealed that sodium nitrite did not fully prevent the impairment of cognitive function because of chronic stress. These data suggest that nitrite supplementation protects against stressed‐induced cerebrovascular dysfunction by limiting the actions of oxidants, potentially via XOR, while improving NO bioavailability. However, nitrite was not sufficient to prevent cognitive impairment with chronic stress.

This study aims to examine the effects of acute whole-body photobiomodulation (wbPBM), applied pre-exercise, on bouts of anaerobic cycling (Wingate) performances. Forty-eight healthy, active males and females participated in this single-blind, randomized, crossover study. Participants visited the laboratory three times to complete repeat (4 ×) Wingate testing, with one week between each visit. All participants completed baseline testing during their first visit and randomly received either the wbPBM or placebo condition before testing on the second visit, followed by the opposite condition on the third visit. There were no significant condition × time interactions for any variable (peak power, average power, power decrement, lactate, heart rate, ratings of perceived exertion, heart rate variability (HRV), root-mean square of differences between R-R intervals (rMSSD), power in the high-frequency range (HF) average, power in the low-frequency range (LF) average, total power, LF/HF, or power in the very-low-frequency range average). A main condition effect was only noted for heart rate, where peak heart rate was significantly higher for wbPBM (145, 141–148 bpm) than placebo (143, 139–146 bpm; p = 0.006) and baseline testing (143, 140–146; p = 0.049) throughout the entire testing session (i.e., collapsed across all timepoints). Furthermore, HRV (rMSSD) the following morning after testing was significantly higher for the wbPBM session compared to placebo ( p = 0.043). There were no differences in perceived recovery ( p = 0.713) or stress ( p = 0.978) scores between wbPBM and placebo. Implementing 20 min of wbPBM immediately prior to maximal bouts of anaerobic cycling did not improve performance (i.e., power output) or physiological responses (e.g., lactate). However, wbPBM elicited the ability to work at a higher heart rate throughout testing and seemed to enhance recovery through improved HRV the following morning.

New FindingsWhat is the central question of this study?Thoracic perivascular adipose tissue (tPVAT) is known to, in part, regulate aortic function: what are the effects of unpredictable chronic mild stress (UCMS) on the tPVAT regulation of aortic function and what is the role of exercise training in alleviating the potential negative actions of UCMS on tPVAT?What is the main finding and its importance?UCMS causes tPVAT to disrupt endothelium‐dependent dilatation, increases inflammatory cytokine production and diminishes tPVAT‐adiponectin. Exercise training proved efficacious in preventing tPVAT‐mediated disruption of aortic function. The data support a tPVAT mechanism through which chronic stress negatively impacts vascular health, which adds to our knowledge of how psychological disorders might increase the risk of cardiovascular disease.AbstractChronic stress is a major risk for cardiovascular disease. Perivascular adipose tissue (PVAT) has been shown to regulate vascular function; however, the impact of chronic stress and the comorbidity of metabolic syndrome (MetS) on thoracic (t)PVAT is unknown. Additionally, aerobic exercise training (AET) is known to combat the pathology of MetS and chronic stress, but the role of tPVAT in these actions is also unknown. Therefore, the purpose of this study was to examine the effects of unpredictable chronic mild stress (UCMS) on the tPVAT regulation of aortic function and the preventative effect of AET. Lean (LZR) and obese (OZR) Zucker rats (16–17 weeks old) were exposed to 8 weeks of UCMS with and without treadmill exercise (AET). In LZR, UCMS impaired aortic endothelium‐dependent dilatation (EDD) (assessedex vivoby wire myography) and aortic stiffness (assessed by elastic modulus) with no change in OZR subject to UCMS. However, both LZR and OZR UCMS tPVAT impaired EDD compared to respective controls. LZR and OZR subject to UCMS had higher oxidative stress production, diminished adiponectin and impaired aortic nitric oxide levels. Divergently, UCMS induced greater inflammatory cytokine production in LZR UCMS tPVAT, but not in OZR UCMS tPVAT. AET prevented the tPVAT impairment of aortic relaxation with UCMS in LZR and OZR. Additionally, AET reduced aortic stiffness in both LZR and OZR. These beneficial effects on tPVAT regulation of the aorta are likely due to AET preservation of adiponectin, reduced oxidative stress and inflammation, and enhanced nitric oxide. UCMS impaired tPVAT‐regulated aortic function in LZR, and augmented MetS‐induced EDD in OZR. Conversely, AET in combination with UCMS largely preserved aortic function and the tPVAT environment, in both groups.

IntroductionElectronic cigarettes (E‐cigs) are gaining popularity in the United States and worldwide. Little is currently known about the effects of chronic E‐cigarette exposure on cytokine expression throughout the body.MethodsC57BL/6 female mice were randomly assigned to cappuccino flavored E‐vapor (18 mg/ml nicotine, N=12) or filtered air (N=13) exposed groups. Animals were exposed 4 hours per day, 5 days per week for 8 months. After exposure major organs including lung, serum, and gastrocnemius were removed and flash frozen for cytokine analysis. Lung and skeletal muscle (i.e. gastrocnemius muscle) were homogenized with Tris‐HCL‐ buffer. Total protein from homogenates and serum were analyzed with the Bradford method and equal concentrations of total protein were analyzed using a Meso Scale Discovery U‐Plex kit according to manufacturer's instructions. Cytokines analyzed included: vascular endothelial growth factor (VEGF), Interleukin (IL)‐6, IL‐5, IL‐4, tumor necrosis factor‐ alpha (TNF‐α), IL‐1β, IL‐15, IL‐ 10, interferon gamma (IFN‐γ) and granulocyte‐macrophage colony‐stimulating factor (GM‐CSF).ResultsThe lungs of E‐cig exposed mice had 54% less VEGF compared to air exposed controls. IL‐6 was reduced by 57% in serum, but IL‐6 increased 994% in the skeletal muscle of E‐cig exposed mice compared to air exposed controls. IL‐10 was reduced by 87% in the gastrocnemius muscle of E‐cig exposed mice compared to air exposed control mice. No significant differences in the remaining factors (i.e. IL‐5, IL‐4, TNF‐α, IL‐1β, IL‐15, IFNγ, GM‐CSF) were identified between E‐cig and air groups in any of the other organs/tissues tested.DiscussionThe results of this study have implications for immune system response time and muscle growth/regeneration. IL‐6 is important for initiating immune system responses, decreases in its expression may lead to increased frequency or duration of infections by viruses or bacteria. IL‐10 in muscle has been shown to influence macrophages to increase anabolic capability of the muscle. Decreases in VEGF may indicate altered angiogenic potential, changes in muscle permeability and/or increased potential of apoptosis in the lung. Long‐term exposure to E‐cig vapor results in changes in cytokines and growth factors that may negatively influence immune and angiogenic potential.Support or Funding InformationNIH P20GM103434 (West Virginia IDeA Network for Biomedical Research Excellence) and Marshall‐WVU Health Cooperative AwardThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Apple pomace, which is a waste byproduct of processing, is rich in several nutrients, particularly dietary fiber, indicating potential benefits for diseases that are attributed to poor diets, such as non-alcoholic fatty liver disease (NAFLD). NAFLD affects over 25% of United States population and is increasing in children. Increasing fruit consumption can influence NAFLD. The study objective was to replace calories in standard or Western diets with apple pomace to determine the effects on genes regulating hepatic lipid metabolism and on risk of NAFLD. Female Sprague-Dawley rats were randomly assigned (n = 8 rats/group) to isocaloric diets of AIN-93G and AIN-93G/10% w/w apple pomace (AIN/AP) or isocaloric diets of Western (45% fat, 33% sucrose) and Western/10% w/w apple pomace (Western/AP) diets for eight weeks. There were no significant effects on hepatic lipid metabolism in rats fed AIN/AP. Western/AP diet containing fiber-rich apple pomace attenuated fat vacuole infiltration, elevated monounsaturated fatty acid content, and triglyceride storage in the liver due to higher circulating bile and upregulated hepatic DGAT2 gene expression induced by feeding a Western diet. The study results showed the replacement of calories in Western diet with apple pomace attenuated NAFLD risk. Therefore, apple pomace has the potential to be developed into a sustainable functional food for human consumption.

New Findings What is the central question of this study? How does chronic stress impact cerebrovascular function and does metabolic syndrome accelerate the cerebrovascular adaptations to stress? What role does exercise training have in preventing cerebrovascular changes to stress and metabolic syndrome? What is the main finding and its importance? Stressful conditions lead to pathological adaptations of the cerebrovasculature via an oxidative nitric oxide pathway, and the presence of metabolic syndrome produces a greater susceptibility to stress‐induced cerebrovascular dysfunction. The results also provide insight into the mechanisms that may contribute to the influence of stress and the role of exercise in preventing the negative actions of stress on cerebrovascular function and structure. AbstractChronic unresolvable stress leads to the development of depression and cardiovascular disease. There is a high prevalence of depression with the metabolic syndrome (MetS), but to what extent the MetS concurrent with psychological stress affects cerebrovascular function is unknown. We investigated the differential effect of MetS on cerebrovascular structure/function in rats (16–17 weeks old) following 8 weeks of unpredictable chronic mild stress (UCMS) and whether exercise training could limit any cerebrovascular dysfunction. In healthy lean Zucker rats (LZR), UCMS decreased (28%, P < 0.05) ex vivo middle cerebral artery (MCA) endothelium‐dependent dilatation (EDD), but changes in MCA remodelling and stiffness were not evident, though cerebral microvessel density (MVD) decreased (30%, P < 0.05). The presence of UCMS and MetS (obese Zucker rats; OZR) decreased MCA EDD (35%, P < 0.05) and dilatation to sodium nitroprusside (20%, P < 0.05), while MCA stiffness increased and cerebral MVD decreased (31%, P < 0.05), which were linked to reduced nitric oxide and increased oxidative levels. Aerobic exercise prevented UCMS impairments in MCA function and MVD in LZR, and partly restored MCA function, stiffness and MVD in OZR. Our data suggest that the benefits of exercise with UCMS were due to a reduction in oxidative stress and increased production of nitric oxide in the cerebral vessels. In conclusion, UCMS significantly impaired MCA structure and function, but the effects of UCMS were more substantial in OZR vs. LZR. Importantly, aerobic exercise when combined with UCMS prevented the MCA dysfunction through subtle shifts in nitric oxide and oxidative stress in the cerebral microvasculature.

Vasomotor response is related to the capacity of the vessel to maintain vascular tone within a narrow range. Two main control mechanisms are involved: the autonomic control of the sympathetic neural drive (global control) and the endothelial smooth cells capacity to respond to mechanical stress by releasing vasoactive factors (peripheral control). The aim of this study was to evaluate the effects of respiratory muscle training (RMT) on vasomotor response, assessed by flow-mediated dilation (FMD) and heart rate variability, in young healthy females. The hypothesis was that RMT could enhance the balance between sympathetic and parasympathetic neural drive and reduce vessel shear stress. Thus, twenty-four women were randomly assigned to either RMT or SHAM group. Maximal inspiratory mouth pressure and maximum voluntary ventilation were utilized to assess the effectiveness of the RMT program, which consisted of three sessions of isocapnic hyperventilation/ week for eight weeks, (twenty-four training sessions). Heart rate variability assessed autonomic balance, a global factor regulating the vasomotor response. Endothelial function was determined by measuring brachial artery vasodilation normalized by shear rate (%FMD/SR). After RMT, but not SHAM, maximal inspiratory mouth pressure and maximum voluntary ventilation increased significantly (+31% and +16%, respectively). Changes in heart rate variability were negligible in both groups. Only RMT exhibited a significant increase in %FMD/SR (+45%; p<0.05). These data suggest a positive effect of RMT on vasomotor response that may be due to a reduction in arterial shear stress, and not through modulation of sympatho-vagal balance.

Proponents for electronic cigarettes (E-cigs) claim that they are a safe alternative to tobacco-based cigarettes; however, little is known about the long-term effects of exposure to E-cig vapor on vascular function. The purpose of this study was to determine the cardiovascular consequences of chronic E-cig exposure. Female mice (C57BL/6 background strain) were randomly assigned to chronic daily exposure to E-cig vapor, standard (3R4F reference) cigarette smoke, or filtered air ( n = 15/group). Respective whole body exposures consisted of four 1-h-exposure time blocks, separated by 30-min intervals of fresh air breaks, resulting in intermittent daily exposure for a total of 4 h/day, 5 days/wk for 8 mo. Noninvasive ultrasonography was used to assess cardiac function and aortic arterial stiffness (AS), measured as pulse wave velocity, at three times points (before, during, and after chronic exposure). Upon completion of the 8-mo exposure, ex vivo wire tension myography and force transduction were used to measure changes in thoracic aortic tension in response to vasoactive-inducing compounds. AS increased 2.5- and 2.8-fold in E-cig- and 3R4F-exposed mice, respectively, compared with air-exposed control mice ( P < 0.05). The maximal aortic relaxation to methacholine was 24% and 33% lower in E-cig- and 3R4F-exposed mice, respectively, than in controls ( P < 0.05). No differences were noted in sodium nitroprusside dilation between the groups. 3R4F exposure altered cardiac function by reducing fractional shortening and ejection fraction after 8 mo ( P < 0.05). A similar, although not statistically significant, tendency was also observed with E-cig exposure ( P < 0.10). Histological and respiratory function data support emphysema-associated changes in 3R4F-exposed, but not E-cig-exposed, mice. Chronic exposure to E-cig vapor accelerates AS, significantly impairs aortic endothelial function, and may lead to impaired cardiac function. The clinical implication from this study is that chronic use of E-cigs, even at relatively low exposure levels, induces cardiovascular dysfunction. NEW & NOTEWORTHY Electronic cigarettes (E-cigs) are marketed as safe, but there has been insufficient long-term exposure to humans to justify these claims. This is the first study to report the long-term in vivo vascular consequences of 8 mo of exposure to E-cig vapor in mice (equivalent to ~25 yr of exposure in humans). We report that E-cig exposure increases arterial stiffness and impairs normal vascular reactivity responses, similar to other risk factors, including cigarette smoking, which contribute to the development of cardiovascular disease.

Nicotine and cigarette smoking is known to promote weight loss and suppress appetite. Since becoming available in 2007, the use of electronic cigarettes (E‐cig) has increased dramatically in the US, however there are still very few studies that examine the long‐term consequences of e‐vapor, particularly in the context of appetite regulation or weight management. The present study was devised to compare the effects of cigarette smoke and E‐cig vapor exposure on food intake, body weight, and body composition in mice. We hypothesized that E‐cigs would elicit similar changes on body mass, adiposity, and food intake as conventional cigarettes (i.e. 3R4F reference cigarette). Female C57BL/6 mice were exposed to filtered room air (n=15), mainstream smoke from 3R4F reference cigarette (n=15), or cappuccino E‐cig vapor (n=15). We report assessments in body mass, food intake, and body composition following daily exposure (4 h/d, 5d/wk) for up to 6 months. Food and water were administered ad libitum. 3R4F, E‐cig and control mice increased body mass by 15%, 30%, and 31%, respectively, over 6 months. 3R4F mice had 15–16% lower body mass (24.9±0.55 g, p<0.01) compared to E‐cig (28.2±0.77 g) and controls (27.7±0.72 g). 3R4F exposed mice also exhibited reduced total body fat (2.74±0.25 g, p<0.01) compared to E‐cig (4.10±0.42 g) and controls (4.18±0.40 g). E‐cig mice food consumption was significantly increased compared to 3R4F (12.4±0.8 vs 9.5±0.7 g, respectively; p<0.05) but not significantly different compared to controls (12.4±0.8 vs 10.5±0.8 g, respectively, p=0.12). Unlike conventional cigarettes, we found that our E‐cig exposed mice did not elicit reductions in total body or adipose mass. This suggests the effects of E‐cig may not be the same as that occurring with traditional tobacco cigarettes, or that the exposure to nicotine and/or other chemicals in the E‐cig liquid elicits a different response on appetite or feeding behavior. Further studies are needed to evaluate the effect that flavorings and/or the compounds produced in E‐cig vapor exert on metabolism and the hypothalamic appetite neurosystems.Support or Funding InformationNIH Grant P20GM103434 (West Virginia IDeA Network for Biomedical Research Excellence) and Marshall‐WVU Health Initiative Award

Individuals with metabolic syndrome (MetS) have an increased aortic stiffness which is a strong, independent risk factor for cardiovascular disease. Additionally, MetS is associated with a decreased capacity for endothelium‐dependent, nitric oxide (NO)‐induced vasorelaxation which may contribute to the aortic stiffness and increased afterload on the heart. Obese Zucker rats, a translational rodent model of MetS, develop impaired vascular reactivity in aortic rings concomitant to the onset of MetS pathologies. Therapeutic interventions aimed at improving endothelial functioning are therefore considered important avenues for improving CV outcomes in MetS patients. We hypothesized that a chronic aerobic exercise intervention would improve endothelium‐dependent relaxation to methacholine in isolated aortic rings of obese Zucker rats. A total of 28 lean and obese Zucker rats (LZR and OZR, respectively) underwent 8 weeks of a treadmill exercise protocol beginning at 9 weeks of age. The animals were separated into 4 groups: sedentary LZR (LZR; n=7); sedentary OZR (OZR; n=6); exercise LZR (LZR‐EX; n=7); exercise OZR (OZR‐EX; n=7). Following the intervention, the thoracic aorta was dissected out from each animal and sectioned into rings, some of which were then mounted into an ex vivo wire tension myograph system. The remaining rings were evaluated for NO production in a DAF‐FM diacetate assay. Force transduction was used to measure the changes in aortic tension in response to pharmacological agonists. The aortic rings were mechanically set to 1 gram of tension then pre‐constricted using phenylephrine (1×10−6 μM), followed by a gradual dilation induced by increasing concentrations of methacholine (1×10−9, 1×10−8, 1×10−7, 1×106, 1×10−5 μM, respectively). Sedentary LZR demonstrated greater maximum methacholine‐induced vasorelaxation compared to sedentary OZR (83% vs. 67%) as well as reduced production of NO in the DAF assay. The exercise trained groups experienced significant improvements in maximal relaxation compared to their sedentary counterparts (LZR‐EX 93%; OZR‐EX 81%), as well as increased values of NO production measured in the DAF assay. These results demonstrate that 8 weeks of aerobic exercise enhances endothelial bioavailability and/or production of NO which improves endothelium‐dependent vasorelaxation in the aortas of both LZR and OZR. Exercise training may therefore be an important therapy for promoting greater arterial compliance in individuals with MetS.Support or Funding InformationNIHAHA

Individuals with increased levels of chronic stress have an increased aortic stiffness which is a strong, independent risk factor for cardiovascular disease. Additionally, chronic stress is associated with a decreased capacity for endothelium‐dependent, nitric oxide (NO)‐induced vasorelaxation which may contribute to the aortic stiffness and increased after load on the heart. Using the unpredictable chronic mild stress protocol (UCMS), lean zucker rats (LZR; healthy rats) develop impaired vascular reactivity in aortic rings concomitant to the onset of chronic stress pathologies. Therapeutic interventions aimed at improving endothelial functioning are therefore considered important avenues for improving CV outcomes in stressed patients. We hypothesized that a chronic aerobic exercise intervention would improve endothelium‐dependent relaxation to methacholine in isolated aortic rings of stressed LZRs. A total of 22 LZRs underwent 8 weeks of a treadmill exercise protocol beginning at 9 weeks of age. The animals were separated into 4 groups: sedentary LZR (LZR; n=7); UCMS LZR (UCMS‐LZR; n=8); exercise and UCMS (ExUCMS‐LZR n=7). Following the intervention, the thoracic aorta was dissected out from each animal and sectioned into rings, some of which were then mounted into an ex vivo wire tension myograph system. The remaining rings were evaluated for NO production in a DAF‐FM diacetate assay. Force transduction was used to measure the changes in aortic tension in response to pharmacological agonists. The aortic rings were mechanically set to 1 gram of tension then pre‐constricted using phenylephrine (1×10‐6μM), followed by a gradual dilation induced by increasing concentrations of methacholine (1×10‐9, 1×10‐8, 1×10‐7, 1×106, 1×10‐5 μM respectively). The LZR group demonstrated greater methacholine‐induced maximal vasorelaxation compared to the UCMS‐LZR group (83% vs. 78%). The ExUCMS‐LZR group experienced the greatest maximal dilation (90%) as well as increased values of NO production measured in the DAF assay compared to LZR group and UCMS‐LZR group. These results demonstrate that 8 weeks of aerobic exercise enhances endothelial bioavailability and/or production of NO which improves endothelium‐dependent vasorelaxation in the aortas of stressed LZRs even more so than the sedentary control. Exercise training may therefore be an important therapy for promoting greater arterial compliance of chronically stressed individuals.Support or Funding InformationNIHAHA

Several studies have characterized the density of beta‐cell mass in rodents reporting greater amounts in the body and tail (which is most proximal to the spleen) compared to the head of the pancreas (located in curved of the duodenum). Whether or not obesity alters the distributions within the pancreas is not known. PURPOSE The objective of this study was to evaluate islet area and beta‐cell density in a rat model of obesity and metabolic syndrome. METHODS Obese (OZR, n=8) and lean Zucker rats (LZR, n=8) were euthanized at 17‐weeks of age. Pancreases were harvested, sectioned into three regions (head, body, tail) and flash frozen in liquid N 2 cooled isopentane. Frozen 7μm thick sections of pancreas tissues were cut, mounted on glass slides, and incubated with an insulin antibody that was labeled with Alexa‐Fluor 546 to identify the beta cells. The tissue was covered in DAPI to label the cell nuclei. Digital images of the pancreas cells were obtained by a Zeiss LSM 510 Confocal microscope using Zenn 2009 software. The insulin positive area of the stained islets was measured using Image J. The images of stained cells and islets were merged and the number of beta cells per islet recorded. ANOVA was used to compare the insulin positive area (islet area), total beta cell number, and beta cell density (beta cell/μm 2 insulin positive area) between obese and lean rats with significance set at P<0.05. RESULTS There was a significant main effect of phenotype (obese vs lean) for all three dependent variables ( Table 1 ) with overall values lower in the OZR. There was also a significant interaction for phenotype and insulin positive area (islet area) as well as an interaction for phenotype and total beta cell number, but not for beta cell density. This suggests the biggest difference associated with obesity within the pancreas is in the tail region causing a reduced islet area. CONCLUSION These data indicate that obesity can significantly reduce islet area but that its affect is primarily in the tail region of the pancreas. The heterogeneity of islet area is consistent with previous findings among the different regions in the pancreas of the rat. Future research needs to examine what mechanisms are responsible for this regional difference. Support or Funding Information NIH AHA Mean±SE Group (n=8) Mean Islet Area (μm 2 ) Total Beta Cells Beta cell/μm 2 islet area OZR LZR OZR LZR OZR LZR Head 6859 ±859 8266 ±266 82±26 115±15 0.0124 ± 0.0008 0.0144 ± 0.0008 Body 7831 ±831 7335 ±335 94±4 98±83 0.0123 ± 0.0010 0.0139 ± 0.0008 Tail 4749 ±651 * 9475 ±475 56 ±6 136 ±3 0.0118 ± 0.0007 0.0147 ± 0.0004

Metabolic syndrome (MetS) is a major risk factor of cardiovascular disease and exercise is recognized for preventing and reversing cardiovascular impairments. Evidence suggests perivascular adipose tissue (PVAT) plays an active roll in vascular function and structure. PVAT is known to release vasoactive substances such as nitric oxide (NO), which may contribute directly to vascular tone. In disease states PVAT may undergo whitening, shifting its phenotype from brown to white reflected by decreased expression of uncoupling protein‐1 (UCP1). Obesity is associated with this whitening of PVAT and a loss of beneficial actions. Further inflammatory markers (IL‐6, and TNF α) along with reactive oxygen species (ROS) released from PVAT may effect aortic endothelium's production of NO. These same cytokines may affect the expression of extracellular matrix remodeling proteins (MMP 2 and 9, and their inhibitors TIMPs), which may alter aortic stiffness. The purpose of the current study is to determine how the presence of MetS alters PVAT phenotype (UCP1), NO and matrix remodeling expression, and to what extent exercise training can affect PVAT expression of above substances. Aortic PVAT was obtained from obese Zucker rats (OZR) a model of MetS, and their lean counterparts (LZR), who either remained inactive or underwent treadmill running for 8 weeks. Using qPCR, PVAT mRNA expression was examined. OZR had a whitening of PVAT shown by a 103‐fold decrease in UCP1. This whitening of PVAT leading to a robust increase in inflammatory cytokine expression (IL‐6 390,000 fold and TNF α 74 fold), with a minor decrease in SOD1 (1.3fold). Such changes may leave the PVAT ill equipped to deal with the increase of ROS associated with MetS. These changes are accompanied by MetS up‐regulation of eNOS (1.8 fold increase, p≤ 0.001) and a decrease in Gch1 (1.6 fold, p≤ 0.001) expression in OZR, suggesting coupling of eNOS may be disrupted. The increase in inflammation and decrease in antioxidant system may also promote increases in remodeling factors. Indeed, MetS caused an increase in both MMP2 (2.3 fold) and MMP9 (6.6 fold) expression, accompanied by a decrease (1.6 fold) in the expression of the inhibitor TIMP1. Foremost exercising led to re‐browning of PVAT with UCP1 up‐regulated 29 fold in OZR. This was followed by improved PVAT SOD1 (3.2 fold p≤ 0.05) expression and a decreased production of IL‐6 (22 fold p≤ 0.05) and TNF α (2.1 fold). Improving the local aortic environment with decreased ROS and inflammation with exercise appeared to balance expression of eNOS (1.2 fold decrease) and Gch1 (2.3 fold increase) suggesting better eNOS coupling and function. Exercise also reduced MMP‐2 (1.3 fold) and MMP9 (8 fold), possible showing beneficial effects of exercise lowering elastase production in PVAT. In conclusion, MetS causes a whitening of PVAT, increased inflammation, decrease in SOD1. These changes may alter the PVAT environment promoting arterial remodeling and increased vascular tone. Exercise combated these MetS impairments by a re‐browning of the PVAT, improving the antioxidant system, reducing inflammation, improving the BH 4 pathway, and decreasing MMPs. Support or Funding Information NIH AHA

Chronic stress/depression is a major risk factor for cardiovascular disease (CVD) and impaired vascular function. In animals with comorbid metabolic syndrome, we have previously shown that chronic stress/depression will cause the near‐total loss of vascular reactivity in cerebral blood vessels, significantly impairing regulation of cerebral blood flow. The purpose of this study was twofold: first, to further elucidate mechanical and anatomical effects of chronic stress in the cerebrovasculature of lean and obese Zucker rats (LZR, OZR), and secondly, to determine the efficacy of exercise training as a prophylactic method for abrogating the onset of stress‐induced vasculopathy in LZR and OZR. Eight weeks of an unpredictable chronic mild stress (UCMS) protocol was imposed on 9 week old male LZR and OZR to induce depressive behaviors. Half of all animals also underwent 8 weeks of daily treadmill exercise concurrent to the UCMS protocol. After sacrifice, the brains were removed and the middle cerebral artery (MCA) was isolated and cannulated in an ex vivo microvessel preparation. Vascular reactivity was assessed to acetylcholine (ACh) on naïve vessels, and again following acute incubation with TEMPOL. The brains were frozen in Tissue Freezing Medium, sectioned, and analyzed for microvessel density (MVD). Plasma samples were analyzed by ELIZA or Meso Scale Diagnostics for biomarkers of inflammation and stress. Vascular reactivity was impaired in MCAs from both UCMS‐LZR and OZR compared to non‐stressed control. OZR‐UCMS demonstrated an almost total loss of reactivity to ACh, which was rescued marginally by TEMPOL. Both LZR‐UCMS and OZR control animals showed elevated levels of inflammatory biomarkers and corticosterone compared to LZR control, and OZR also had a significant decrease in cortical MVD. A smaller, but significant, decrease in MVD was also observed in LZR‐UCMS, while an ever larger drop in cortical MVD was seen in OZR‐UCMS. Exercise training was able to completely reverse the effects of UCMS in both LZR and OZR; LZR‐UCMS‐Exercise was comparable to LZR control, while OZR‐UCMS‐Exercise showed improved reactivity beyond OZR control. Interestingly, TEMPOL treatment of vessels from exercised animals resulted in no significant further improvements in reactivity. Plasma inflammatory biomarkers were significantly decreased in exercise trained animals, and the decreases in cortical MVD were prevented as well, with OZR exercise animals actually showing an increase over control. These results suggest that exercise training can protect the cerebrovasculature from the detrimental effects of chronic stress/depression, likely due to the anti‐oxidant and anti‐inflammatory benefits of exercise. Enhanced clearance of ROS and improved NO bioavailability helps improve vascular function and prevent rarefaction of cortical microvessels, indicating that these pathways may be promising avenues for treating stress‐related cerebrovascular disease. Support or Funding Information American Heart Association Predoctoral Fellowship NIH COBRE grant

Chronic stress has been implicated as a possible contributing factor to the onset of Type 2 Diabetes Mellitus through its negative effects on the insulin producing β cells of the pancreas. The objective of this study was to evaluate a model of chronic stress and the metabolic syndrome on beta cell density in rats. Obese (OZR) and lean Zucker rats (LZR) were randomly divided into stress (OZR‐S, n=6; LZR‐S, n=6) and non‐stress (OZR‐NS, n=3; LZR‐NS, n=3). Stress included bath, damp bedding, no bedding, cage tilt, altered light cycles, and cage‐switching for 7 hours/day, 5 days/week for 8 weeks after which animals were euthanized. Pancreases were harvested and mounted on cork using OTC freezing medium. Pancreas sections of 7µm thickness were cut and collected on slides then stained using Alexa‐Fluor 546 targeting primary insulin antibodies and DAPI. Slides were then imaged using the Zeiss LSM 510 Confocal microscope and Zeiss computer software. The insulin positive area of the stained islets was measured using the Image J program.The images of stained cells and islets were merged and the number of beta cells per islet recorded. A two‐way ANOVA was used to compare the beta cell density (beta cell/µm 2 insulin positive area) among groups with significance set at p<.05. There was a significant main effect of stress with stressed animals having a lower beta cell density compared with non‐stressed (Stressed: 0.00566±.001; Non‐Stressed: 0.00861±.001µm 2 ). No differences were observed between OZR and LZR. These results indicate that 8 weeks of chronic stress caused beta cell density to be reduced in both OZR and LZR and this was not affected further by the metabolic condition of the animals.

PURPOSE:Reducing vascular endothelial growth factor (VEGF) in adipose tissue alters adipose vascularity and metabolic homeostasis. We hypothesized that this would also affect metabolic responses during exercise-induced stress and that adipocyte-specific VEGF-deficient (adipoVEGF-/-) mice would have impaired endurance capacity. METHODS:Endurance exercise capacity in adipoVEGF-/- (n = 10) and littermate control (n = 11) mice was evaluated every 4 wk between 6 and 24 wk of age using a submaximal endurance run to exhaustion at 20 m·min(-1) at 10° incline. Maximal running speed, using incremental increases in speed at 30-s intervals, was tested at 25 and 37 wk of age. RESULTS:White and brown adipose tissue capillarity were reduced by 40% in adipoVEGF-/-, and no difference in skeletal muscle capillarity was observed. Endurance run time to exhaustion was 30% lower in adipoVEGF-/- compared with that in controls at all time points (P < 0.001), but no difference in maximal running speed was observed between the groups. After exercise (1 h at 50% maximum running speed), adipoVEGF-/- mice displayed lower circulating insulin (P < 0.001), lower glycerol (P < 0.05), and tendency for lower blood glucose (P = 0.06) compared with controls. There was no evidence of altered oxidative damage or changes in carnitine palmitoyltransferase-1β expression in skeletal muscle of adipoVEGF-/- mice. CONCLUSIONS:These data suggest that VEGF-mediated deficits in adipose tissue blunt the availability of lipid substrates during endurance exercise, which likely reduced endurance performance. Surprisingly, we also found an unchanged basal blood glucose despite lower circulating insulin in adipoVEGF-/- mice, suggesting that loss of adipocyte VEGF can blunt insulin release and/or increase basal insulin sensitivity.

The purpose of this study was to investigate the effects of one extra hour of sleep on physical performance and nutrition behaviors on college female track athletes. Eleven females (age 20.6 ± 1.6 yrs) maintained normal sleep habits for a one week. Baseline data was followed by a one week sleep extension period (night time and napping). Anaerobic power and fatigue was measured using the Wingate Test. The ASA 24‐hour food‐recall questionnaire was used to test nutrient intake. Reaction time, perceived stress, and mood were measured using the Psychomotor Vigilance Task (PVT), Cohen's Stress Test, and Profile of Mood States (POMS), respectively. Preliminary data was collected on 11 female subjects. Subjects increased total sleep from baseline (423.5 ± 44.4 minutes, 443.9 ± 55.2 minutes respectively), however this was not significant (z = ‐1.2480, p = 0.2258). No significant differences were seen in maximum power (z = 0.00, p = 1.0000) and fatigue (z = 0.0657, p = 0.9476) following sleep extension. Although not significant, mean PVT reaction time increased (z = ‐0.8213, p = 0.4115) and POMS scores improved (z = 0.7560, p = 0.4496) following more sleep. The athletes showed no significant effects in caloric (z = 0.1313, p = 0.8968) or total fat intake (z = ‐0.2627, p = 0.7928) after sleep extension. Improvements in physical performance and nutritional behaviors were not seen after sleep extension in college female track athletes.Grant Funding Source: WVU Experimental Station Hatch Funding

Context. Hydration is important for all athletes not only for overall health, but for optimal performance during competition. Many studies have shown a relationship between a decrease in physiological function and performance as related to dehydration typically by measuring peak power output and aerobic capacity. But few studies have shown the relationship between hydration or dehydration and the performance of wrestlers throughout a competitive season. Objective. The purposes of this study were to determine: (1) if there is a difference between hydration levels over three time points between a high specific gravity group and a low specific gravity group; (2) if those who have the lower average specific gravity have a higher win percentage and; (3) if those who have a lower average specific gravity earn more team points throughout a competitive season. Design. This study was a longitudinal prospective comparative study. The independent variables for the first design were the three time points measured; baseline, 24-hours pre-competition (24PC), and pre-competition (PC), and a high average specific gravity group (HSG, >1.02) and a low average specific gravity group (LSG, ≤1.02). The dependent variable was the specific gravity. The second design independent variable was based on two groups: HSG and LSG. The dependent variable was win percentage. The independent variable for the third purpose was based on two groups as well: HSG and LSG. The dependent variable was team points earned by the wrestler. Setting. The study took place at a Division AAA high school in Western Pennsylvania. Patients or other participants . Voluntary high school wrestlers between the ages of 14 and 18 (16.53+/-1.41 yrs) and grades 9--12 were used for the study. Subjects were excluded from the study if they did not participate in at least 75% of matches throughout the competitive season. Interventions. Specific gravity of urine and body weight was measured at baseline, 24 hours before a competition and then again the day of the competition, 2--3 hours before the start of the first match for home matches. For away competitions specific gravity was measured before the bus left for the match. Win percentages were calculated from dual meets as well as the first day of tournaments. Team points based on performance were measured on the outcome of each individuals match during dual meets as well as the first day of tournaments. Main outcome measures . The investigator believed that specific gravity would change over time with the LSG showing less variability than HSG. Those athletes in the LSG (i.e. more hydrated) will show a higher win percentage and earn more team points during those matches. Results. There was a significant time by group interaction (P=.005), for the HSG at baseline (P=.045) and HSG at 24PC (P=.004). A main effect for time (P<.001) for all 3 time points: baseline to 24PC (P<.001), baseline to PC (P<.001), and 24PC to PC (P=.041). LSG had an average team point total of 14.33+/-7.74. HSG had an average team point total of 32+/-14.34. There was a significant difference between the two groups (P=.024) with HSG earning more team points. All other results showed no significance. Conclusions. The results of this study show a group by time interaction at baseline and 24PC with HSG being lower at baseline but higher at 24PC. A main effect for time showed specific gravity does change significantly from baseline to 24PC, baseline to PC, and 24PC to PC, and is significantly higher 24PC and PC than baseline. Also there was a significant difference between the groups for team points. However, HSG scored significantly more team points than LSG. There was no significant difference measured between groups when compared to win percentages.

Background. Accumulation of free fatty acids leads to lipid-toxicity-associated skeletal muscle atrophy. Palmitate treatment reduces myoblast and myotube growth and causes apoptosis in vitro. It is not known if omega-3 fatty acids will protect muscle cells against palmitate toxicity. Therefore, we examined the effects of docosahexaenoic acid (DHA) on skeletal muscle growth. Methods. Mouse myoblasts (C2C12) were differentiated to myotubes, and then treated with 0 or 0.5 mM palmitic acid or 0 or 0.1 mM DHA. Results. Intramyocellular lipid was increased in palmitate-treated cells but was prevented by DHA-palmitate cotreatment. Total AMPK increased in DHA+ palmitate-treated compared to palmitate only cells. RpS6 phosphorylation decreased after palmitate (-55%) and this was blunted by DHA+ palmitate (-35%) treatment. Palmitate treatment decreased PGC1α protein expression by 69%, but was increased 165% with DHA+ palmitate (P = 0.017) versus palmitate alone. While palmitate induced 25% and 90% atrophy in myotubes (after 48 hours and 96 hours, resp.), DHA+ palmitate treatment caused myotube hypertrophy of ~50% and 100% after 48 and 96 hours, respectively. Conclusion. These data show that DHA is protective against palmitate-induced atrophy. Although DHA did not activate the AMPK pathway, DHA treatment restored growth-signaling (i.e., rpS6) and rescued palmitate-induced muscle atrophy.