Effects of Acute Cold Exposure on Plasma Biomarkers Associated with Cardiovascular Disease

Inflammatory cytokines and lipid mediators are used as biomarkers for CVD risk. Cold exposure has been suggested to improve some of these biomarkers. We measured cardiovascular inflammatory and lipid biomarkers to expand our knowledge of cold exposure and CVD risk. Interleukin-1 Beta (IL-1 B) and Chemokine Ligand 2 (CCL2) are inflammatory cytokines associated with various disease states. Free fatty acids (FFA) are released from fat cells in response to stress. We evaluated the effects of acute 30 min cold exposure on these blood biomarkers. We hypothesized that the inflammatory markers and plasma FFA levels would increase at 2-h post-cold exposure. Twenty subjects (9 females, 23.9±2.7sd y, 1.71±10.2sd m, 74.2±13.5sd kg, 19.4±7.4sd %BF, 64.5±15.3sd kg FFM) were subjected to a 30-min seated cold exposure while metabolic data was collected via indirect calorimetry. Shivering started immediately upon cold exposure and ceased within 10 seconds following cold exposure. Estimated resting energy expenditure (kcal/min) during the exposure period (1.73±0.7sd, 1.47±0.6sd, 1.36±1.0sd for min 5, 15, and 30, respectively) was two-fold higher than pre-cold. Venous plasma was collected at pre-cold period, immediately after cold exposure, and 2 h post-cold, centrifuged, and stored at -80 ºC for subsequent biomarker analysis. ELISAs were used to measure plasma inflammatory cytokines; interlukin-1 beta (IL-1 B) and chemokine ligand 2 (CCL2) biomarkers and plasma free fatty acid (FFA) during and following cold exposure. Pre-cold IL-1B (pg/ml), CCL2 (pg/ml), and FFA (mM) values were 19±3.6se, 2.4±0.7se, and 507±87se, respectively. Immediate post-cold values were 18±3.8se, 3.4±1.2se, and 412±42se, respectively. Finally, 2 h post-cold values were 20±3.6se, 2.6±0.7se, and 458±48se, respectively. There was a 3.4±0.7se (p\u3c0.05) increase in IL-1 B plasma levels immediately after cold exposure that lasted up to two hours. CCL2 plasma levels and FFA were not different from baseline during the post-cold period. We conclude that acute cold exposure may worsen CVD risk through a select inflammatory response. Additional analyses of our samples will expand the possible list of affected CVD risk biomarkers. Whether or not extended exposure to cold would exacerbate these marker levels or affect the other markers measured is not known

Effects of Extra Virgin Olive Oil Phytochemicals Supplementation and Aerobic Exercise Training on Inflammatory and Hormonal Markers in Rats Fed Atherogenic Diet

Westernized diet and sedentary lifestyle may exacerbate systemic inflammation and provoke hormonal changes. Extra virgin olive oil (EVOO) derived phytochemicals Oleocanthal (Oleo) and Oleacein (Olea) demonstrate anti-inflammatory and antioxidant effects. PURPOSE: The purpose of the study is to evaluate the effects of Oleo/Olea supplementation and aerobic exercise training (AET) on plasma inflammatory cytokines and hormones in rats fed high fat atherogenic diet. METHODS: Forty-eight female Sprague-Dawley rats were fed an atherogenic diet consisting of 1.25% cholesterol and 0.5% cholic acid for 12 weeks. Half of the rats were supplemented with 20% EVOO containing high Oleo/Olea (1000 mg/kg, HO, n=24) or low Oleo/Olea (100 mg/kg, LO, n=24). In each dietary group, half group was trained (T) on treadmill for 12 weeks (25m/min, 10% grade for 60 min/day, 5 days/week), while the other half remained sedentary. Two separate groups of rats were fed a chow diet (C, n=6) and atherogenic diet (A, n=12), respectively without EVOO or T. Inflammatory cytokines (interleukin (IL)-1β, monocyte chemoattractant protein (MCP)-1 and tumor necrosis factor (TNF)-α) and hormones (leptin and insulin) were measured in plasma using Bioplex 200 Multiplex immunoassay. RESULTS: IL-1β and TNF-α were increased in A vs. C (P=\u3c0.01). LO increased IL-1β compared to A (P\u3c0.05) but not HO. HO showed a trend to lower IL-1β vs.LO (P=0.064). Although MCP-1 and TNF-α were both decreased in EVOO vs. A (P\u3e0.05), significant differences were only found in EVOO+T compared to A (P\u3c0.05). Insulin was also decreased in LO vs. A (P\u3c0.05), but elevated in HO vs. LO (P\u3c0.05). CONCLUSION: High-fat atherogenic diet induced plasma inflammatory cytokines but were ameliorated by EVOO supplementation and AET. Hormones leptin and insulin were decreased by EVOO supplementation and AET

Acute maternal oxidant exposure causes susceptibility of the fetal brain to inflammation and oxidative stress

Background Maternal exposure to environmental stressors poses a risk to fetal development. Oxidative stress (OS), microglia activation, and inflammation are three tightly linked mechanisms that emerge as a causal factor of neurodevelopmental anomalies associated with prenatal ethanol exposure. Antioxidants such as glutathione (GSH) and CuZnSOD are perturbed, and their manipulation provides evidence for neuroprotection. However, the cellular and molecular effects of GSH alteration in utero on fetal microglia activation and inflammation remain elusive. Methods Ethanol (EtOH) (2.5 g/kg) was administered to pregnant mice at gestational days 16–17. One hour prior to ethanol treatment, N-acetylcysteine (NAC) and L-buthionine sulfoximine (BSO) were administered to modulate glutathione (GSH) content in fetal and maternal brain. Twenty-four hours following ethanol exposure, GSH content and OS in brain tissues were analyzed. Cytokines and chemokines were selected based on their association with distinctive microglia phenotype M1-like (IL-1β, IFN γ, IL-6, CCL3, CCL4, CCL-7, CCL9,) or M2-like (TGF-β, IL-4, IL-10, CCL2, CCL22, CXCL10, Arg1, Chi1, CCR2 and CXCR2) and measured in the brain by qRT-PCR and ELISA. In addition, Western blot and confocal microscopy techniques in conjunction with EOC13.31 cells exposed to similar ethanol-induced oxidative stress and redox conditions were used to determine the underlying mechanism of microglia activation associated with the observed phenotypic changes. Results We show that a single episode of mild to moderate OS in the last trimester of gestation causes GSH depletion, increased protein and lipid peroxidation and inflammatory responses inclined towards a M1-like microglial phenotype (IL-1β, IFN-γ) in fetal brain tissue observed at 6–24 h post exposure. Maternal brain is resistant to many of these marked changes. Using EOC 13.31 cells, we show that GSH homeostasis in microglia is crucial to restore its anti-inflammatory state and modulate inflammation. Microglia under oxidative stress maintain a predominantly M1 activation state. Additionally, GSH depletion prevents the appearance of the M2-like phenotype, while enhancing morphological changes associated with a M1-like phenotype. This observation is also validated by an increased expression of inflammatory signatures (IL-1β, IFN-γ, IL-6, CCL9, CXCR2). In contrast, conserving intracellular GSH concentrations eliminates OS which precludes the nuclear translocation and more importantly the phosphorylation of the NFkB p105 subunit. These cells show significantly more pronounced elongations, ramifications, and the enhanced expression of M2-like microglial phenotype markers (IL-10, IL-4, TGF-β, CXCL10, CCL22, Chi, Arg, and CCR2). Conclusions Taken together, our data show that maintaining GSH homeostasis is not only important for quenching OS in the developing fetal brain, but equally critical to enhance M2 like microglia phenotype, thus suppressing inflammatory responses elicited by environmental stressors

The Effect Of The COVID-19 Pandemic And Quarantine Period On The Physical Activity And Dietary Habits Of College-aged Students

BACKGROUND: The COVID-19 pandemic has led to sudden extreme changes in lifestyle, potentially causing adverse changes in physical activity (PA), sedentary behavior (SB), and dietary habits. It is important to investigate how college students are being impacted by the quarantine periods and campus closures. The objective of the study was to investigate the effects of COVID-19 quarantine on frequency of physical activity, frequency of food consumption and food insecurity among college students. The findings of the study will provide preliminary evidence on dietary, PA, and SB changes induced by the pandemic among college students. METHODS: Participants (N = 403) completed a cross-sectional self-report online questionnaire, evaluating the PA, SB, and dietary behaviors before- and during-campus closures (March – May 2020), due to COVID-19. Sociodemographic and descriptive information were also obtained from each participant. A combination of nonparametric statistical tests were used to assess changes in PA, SB, and dietary habits, and between group differences. RESULTS: A total of 291 participants met the inclusion criteria (college-aged, 18-24 years). A mean decrease of 1.26, 1.70, 1.57 days/week (p \u3c .001) were found for vigorous, moderate, and light-intensity PA, respectively. SB increased by 2.66 hours/day, p \u3c0.001. CONCLUSION: College-aged students in quarantine significantly decreased PA levels and increased SB. A significant number of students reported negative changes in dietary habits, most notably in alcohol consumption. Food insecurity significantly increased among the current sample. Public health strategies to attenuate the negative changes in lifestyle habits should be implemented during a global pandemic

Acute maternal oxidant exposure causes susceptibility of the fetal brain to inflammation and oxidative stress

Background: Maternal exposure to environmental stressors poses a risk to fetal development. Oxidative stress (OS), microglia activation, and inflammation are three tightly linked mechanisms that emerge as a causal factor of neurodevelopmental anomalies associated with prenatal ethanol exposure. Antioxidants such as glutathione (GSH) and CuZnSOD are perturbed, and their manipulation provides evidence for neuroprotection. However, the cellular and molecular effects of GSH alteration in utero on fetal microglia activation and inflammation remain elusive. Methods: Ethanol (EtOH) (2.5 g/kg) was administered to pregnant mice at gestational days 16–17. One hour prior to ethanol treatment, N-acetylcysteine (NAC) and L-buthionine sulfoximine (BSO) were administered to modulate glutathione (GSH) content in fetal and maternal brain. Twenty-four hours following ethanol exposure, GSH content and OS in brain tissues were analyzed. Cytokines and chemokines were selected based on their association with distinctive microglia phenotype M1-like (IL-1β, IFN γ, IL-6, CCL3, CCL4, CCL-7, CCL9,) or M2-like (TGF-β, IL-4, IL-10, CCL2, CCL22, CXCL10, Arg1, Chi1, CCR2 and CXCR2) and measured in the brain by qRT-PCR and ELISA. In addition, Western blot and confocal microscopy techniques in conjunction with EOC13.31 cells exposed to similar ethanol-induced oxidative stress and redox conditions were used to determine the underlying mechanism of microglia activation associated with the observed phenotypic changes. Results: We show that a single episode of mild to moderate OS in the last trimester of gestation causes GSH depletion, increased protein and lipid peroxidation and inflammatory responses inclined towards a M1-like microglial phenotype (IL-1β, IFN-γ) in fetal brain tissue observed at 6–24 h post exposure. Maternal brain is resistant to many of these marked changes. Using EOC 13.31 cells, we show that GSH homeostasis in microglia is crucial to restore its anti-inflammatory state and modulate inflammation. Microglia under oxidative stress maintain a predominantly M1 activation state. Additionally, GSH depletion prevents the appearance of the M2-like phenotype, while enhancing morphological changes associated with a M1-like phenotype. This observation is also validated by an increased expression of inflammatory signatures (IL-1β, IFN-γ, IL-6, CCL9, CXCR2). In contrast, conserving intracellular GSH concentrations eliminates OS which precludes the nuclear translocation and more importantly the phosphorylation of the NFkB p105 subunit. These cells show significantly more pronounced elongations, ramifications, and the enhanced expression of M2-like microglial phenotype markers (IL-10, IL-4, TGF-β, CXCL10, CCL22, Chi, Arg, and CCR2). Conclusions: Taken together, our data show that maintaining GSH homeostasis is not only important for quenching OS in the developing fetal brain, but equally critical to enhance M2 like microglia phenotype, thus suppressing inflammatory responses elicited by environmental stressors

Ursolic Acid and Related Analogues: Triterpenoids with Broad Health Benefits

Ursolic acid (UA) is a well-studied natural pentacyclic triterpenoid found in herbs, fruit and a number of traditional Chinese medicinal plants. UA has a broad range of biological activities and numerous potential health benefits. In this review, we summarize the current data on the bioavailability and pharmacokinetics of UA and review the literature on the biological activities of UA and its closest analogues in the context of inflammation, metabolic diseases, including liver and kidney diseases, obesity and diabetes, cardiovascular diseases, cancer, and neurological disorders. We end with a brief overview of UA’s main analogues with a special focus on a newly discovered naturally occurring analogue with intriguing biological properties and potential health benefits, 23-hydroxy ursolic acid.11Ysciescopu

Switching to a Standard Chow Diet at Weaning Improves the Effects of Maternal and Postnatal High-Fat and High-Sucrose Diet on Cardiometabolic Health in Adult Male Mouse Offspring

Cardiac mitochondrial dysfunction contributes to obesity-associated heart disease. Maternal and postnatal diet plays an important role in cardiac function, yet the impacts of a mismatch between prenatal and postweaning diet on cardiometabolic function are not well understood. We tested the hypothesis that switching to a standard chow diet after weaning would attenuate systemic metabolic disorders and cardiac and mitochondrial dysfunction associated with maternal and postnatal high-fat/high-sucrose (HFHS) diet in mice. Six-month-old male CD1 offspring from dams fed a HFHS diet and weaned to the same HFHS diet (HH) or switched to a standard chow diet (HC) were compared to offspring from dams fed a low-fat/low-sucrose diet and maintained on the same diet (LL). HC did not decrease body weight (BW) but normalized glucose tolerance, plasma cholesterol, LDL, and insulin levels compared to the HH. Systolic function indicated by the percent fractional shortening was not altered by diet. In freshly isolated cardiac mitochondria, maximal oxidative phosphorylation-linked respiratory capacity and coupling efficiency were significantly higher in the HC in the presence of fatty acid substrate compared to LL and HH, with modification of genes associated with metabolism and mitochondrial function. Switching to a standard chow diet at weaning can attenuate the deleterious effects of long-term HFHS in adult male mouse offspring

Ursolic acid protects monocytes against metabolic stress-induced priming and dysfunction by preventing the induction of Nox4

Aims: Dietary supplementation with ursolic acid (UA) prevents monocyte dysfunction in diabetic mice and protects mice against atherosclerosis and loss of renal function. The goal of this study was to determine the molecular mechanism by which UA prevents monocyte dysfunction induced by metabolic stress. Methods and results: Metabolic stress sensitizes or “primes” human THP-1 monocytes and murine peritoneal macrophages to the chemoattractant MCP-1, converting these cells into a hyper-chemotactic phenotype. UA protected THP-1 monocytes and peritoneal macrophages against metabolic priming and prevented their hyper-reactivity to MCP-1. UA blocked the metabolic stress-induced increase in global protein-S-glutathionylation, a measure of cellular thiol oxidative stress, and normalized actin-S-glutathionylation. UA also restored MAPK phosphatase-1 (MKP1) protein expression and phosphatase activity, decreased by metabolic priming, and normalized p38 MAPK activation. Neither metabolic stress nor UA supplementation altered mRNA or protein levels of glutaredoxin-1, the principal enzyme responsible for the reduction of mixed disulfides between glutathione and protein thiols in these cells. However, the induction of Nox4 by metabolic stress, required for metabolic priming, was inhibited by UA in both THP-1 monocytes and peritoneal macrophages. Conclusion: UA protects THP-1 monocytes against dysfunction by suppressing metabolic stress-induced Nox4 expression, thereby preventing the Nox4-dependent dysregulation of redox-sensitive processes, including actin turnover and MAPK-signaling, two key processes that control monocyte migration and adhesion. This study provides a novel mechanism for the anti-inflammatory and athero- and renoprotective properties of UA and suggests that dysfunctional blood monocytes may be primary targets of UA and related compounds

Resveratrol and quercetin interact to inhibit neointimal hyperplasia in mice with a carotid injury

Restenosis is a critical complication of angioplasty and stenting. Restenosis is multifactorial, involving endothelial injury, inflammation, platelet activation, and vascular smooth muscle cell (VSMC) proliferation. Thus, dietary strategies to prevent restenosis likely require the use of more than one agent. Resveratrol (R) and quercetin (Q) are polyphenols that are known to exhibit vascular protective effects. We tested whether R and Q administered in the diet interact to inhibit vessel stenosis in mice with a carotid injury. B6.129 mice were administered a high-fat diet containing 21% fat and 0.2% cholesterol along with R (25 mg/kg), Q (10 mg/kg), or R + Q for 2 wk. A carotid injury was induced and the mice were again administered the enriched diet for 2 wk. Compared with the controls, R significantly decreased stenosis, assessed as an intima:media ratio, by 76%. Although Q treatment alone exhibited no effect, it potentiated the effect of R in that treatment with R + Q significantly decreased the intima:media ratio by 94%. Moreover, this effect was greater than that of R treatment alone (P \u3c 0.05). Although treatments with R, Q, and R + Q significantly affected platelet activation and endothelial function, the responses observed for R + Q were less than additive. Specifically, the effects of R + Q were less than the sum of effects for treatments with R and Q alone. In contrast, treatment with R + Q exhibited more-than-additive effects on inflammatory markers and significant interactions between R and Q were observed. The presence of synergy between R and Q was thus tested in cultures of VSMC and macrophages. Isobolographic analysis revealed that 2:1 molar ratios of R:Q exhibited synergistic inhibition of VSMC proliferation and macrophage chemotaxis. In conclusion, in combination, R and Q can interact to reduce the extent of restenosis, perhaps due to their synergistic inhibition of VSMC proliferation and inflammation