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Background: Vitamin E has antioxidant properties, which help in scavenging free radicals, thereby reducing oxidation of lipids and proteins. This study aims to evaluate the efficacy of oral vitamin E supplementation in preventing retinopathy of prematurity (ROP) in very low birth weight (VLBW) infants with respiratory distress syndrome (RDS) and decreasing oxidative stress 15 and 28 days post-intervention. Methods: Ninety VLBW infants were randomly assigned to two groups: 1. Treatment (treatment group (T), n = 48) or 2. Placebo (control group (C), n = 42). Each group received 25 IU of vitamin E (T) or placebo (C). Results: The incidence of ROP in groups T and C was 12.5% (n=6) and 31% (n = 13), respectively (RR: 0.40; 95% CI: 0.10–0.96). There were no differences in mortality between groups. As expected, the vitamin E concentration was significantly increased 28 days post-intervention in group T. Conclusion: Oral supplementation with vitamin E may effectively prevent ROP development in VLBW infants with RDS. Oxidative damage markers were significantly lower, whereas total antioxidant capacity was increased in group T. However, levels of other antioxidants as vitamin A and C were not measured in two groups.

Evidence has implicated oxidative stress (OS) and inflammation as drivers of neurodegenerative pathologies. We previously reported on the beneficial effects of (-)-epicatechin (Epi) treatment on aging-induced OS and its capacity to restore modulators of mitochondrial biogenesis in the prefrontal cortex of 26-month-old male mice. In the present study using the same mouse model of aging, we examined the capacity of Epi to mitigate hippocampus OS, inflammation, hyperphosphorylation of tau protein, soluble β-amyloid protein levels, cell survival, memory, anxiety-like behavior levels and systemic inflammation. Mice were subjected to 4 weeks of Epi treatment (1 mg kg-1 day-1) and samples of the hippocampus were obtained. Assessments of the OS markers, protein carbonyls, and malondialdehyde levels demonstrated their significant increase (∼3 fold) with aging that were partially suppressed by Epi. The protein levels of the glial fibrillary acidic protein, inflammatory factor 1 (Iba1), pro-inflammatory cytokines, interleukins (IL-1β, IL-3, 5, 6 and 15), cyclooxygenase 2, tumor necrosis factor α, nuclear factor-activated B cells and interferon γ increase with aging and were also significantly decreased with Epi treatment. However, anti-inflammatory cytokines, IL-1ra, IL-10 and 11 decrease with aging and were restored with Epi. Epi also reversed the aging effects on the hyperphosphorylation of tau, increased soluble β-amyloid levels (∼2 fold), cellular death (as per caspase 3 and 9 activity), and reduced nerve growth factor and triggering receptor expressed on myeloid cells 2 levels. Measures of anxiety like-behavior and memory demonstrated improvements with Epi treatment. Indicators of systemic inflammation increase with aging and Epi was capable of decreasing blood inflammatory markers. Altogether, the results show a significant capacity of Epi to mitigate hippocampus OS and inflammation leading to improved brain function.

Purpose To compare the retinal sensitivity and evaluate its contribution to the foveal sensitivity in patients with and without diabetes who maintain normal visual acuity. Methods Observational, descriptive, cross-sectional and prospective study in 20 subjects without diabetes (group 1) and 23 with type 2 diabetes mellitus (group 2) that had no ocular abnormalities. Retinal sensitivity was measured with the macular threshold test by the Humphrey's computerized perimeter. The mean sensitivity in each of the 16 points and the foveal sensitivity were compared between groups using the Mann–Whitney's U test; the correlation between retinal sensitivity and foveal sensitivity was analyzed by the Spearman's test and the contribution of each point to the foveal sensitivity was identified by multiple regression. Results Sixty eyes were evaluated, 30 in group 1 and 30 in group 2; the mean foveal sensitivity was 34.77 ± 0.5 dB in group 1 and 32.87 ± 0.6 in group 2. The highest sensitivity of the temporal visual field had an inferior paracentral location (point 3) in both groups. In the linear regression analysis, points which contributed to the foveal sensitivity were 1 in group 1 and points 7 and 15 in group 2. Conclusions Subjects without diabetes have a significantly higher sensitivity in the temporal retina compared with those with diabetes; points with highest mean retinal sensitivity do not correspond to the central four. The reduced sensitivity in point 1 decreases the mean foveal sensitivity in subjects with diabetes, because this variable correlates with lower perimetry points.

In the production of chocolate, only cocoa seeds are used, generating by-products that are generally discarded, increasing the risk of environmental contamination. Given fiber, carbohydrates, proteins, and flavonoid content the use of cacao pod husks can generate nutraceutical products for human consumption. In contrast, obesity represents a major public health problem worldwide. Cacao derivatives are able to modulate overweight and lipid disorders. The objective of present work was to prepare and characterize products using cacao by-products and analyze their effects on altered cardiometabolic risk markers in an obesity model induced by high fat diet and fructose ingestion in rats. The effects of a pellet and extracts made with outer pod husk and kernel husk for 5 weeks were analyzed in an obesity rat model. The treatments significantly decreased body weight by 39%, systolic blood pressure by 27%, triglycerides by 55%, total cholesterol by 24%, low-density lipoprotein cholesterol by 37%, and the triglycerides/high-density lipoprotein ratio by 54%. Cacao by-products improved the metabolic function of obese animals, without causing secondary effects.

KEY POINTS Heart failure with preserved ejection fraction (HFpEF) is seen more frequently in older women; risk factors include age, hypertension and excess weight. No female animal models of early stage remodelling (pre-HFpEF) have examined the effects that the convergence of such factors have on cardiac structure and function. In this study, we demonstrate that ageing can lead to the development of mild chamber remodelling, diffuse fibrosis and loss of diastolic function. The loss of oestrogens further aggravates such changes by leading to a notable drop in cardiac output (while preserving normal ejection fraction) in the presence of diffuse fibrosis that is more predominant in endocardium and is accompanied by papillary fibrosis. Excess weight did not markedly aggravate such findings. This animal model recapitulates many of the features recognized in older, female HFpEF patients and thus, may serve to examine the effects of candidate therapeutic agents. ABSTRACT Two-thirds of patients with heart failure with preserved ejection fraction (HFpEF) are older women, and risk factors include hypertension and excess weight/obesity. Pathophysiological factors that drive early disease development (before heart failure ensues) remain obscure and female animal models are lacking. The study evaluated the intersecting roles of ageing, oestrogen depletion and excess weight on altering cardiac structure/function. Female, 18-month-old, Fischer F344 rats were divided into an aged group, aged + ovariectomy (OVX) and aged + ovariectomy + 10% fructose (OVF) in drinking water (n = 8-16/group) to induce weight gain. Left ventricular (LV) structure/function was monitored by echocardiography. At 22 months of age, animals were anaesthetized and catheter-based haemodynamics evaluated, followed by histological measures of chamber morphometry and collagen density. All aged animals developed hypertension. OVF animals increased body weight. Echocardiography only detected mild chamber remodelling with ageing while intraventricular pressure-volume loop analysis showed significant (P

Cardiac fibrosis is one of the hallmarks of a diabetic cardiomyopathy. When activated, cardiac fibroblasts (CFs) increase the production of extracellular matrix proteins. Transforming growth factor (TGF)

The role of the energy industry has always been central for one reason or another, being environmentalism the main motive in the last two decades. Therefore, attention and research have been directed in this sense. However, human resources—or human capital—have remained understudied, especially concerning the salaries received. Thus, this study is disruptive as it explored the factors that influence employee remuneration in the energy subsector, using Spain as a case study. For this, the PLS-SEM (Partial Least Squares Structural Equation Modelling) path modelling methodology was used, executing a traditional PLS analysis, bootstrapping and, finally, IPMA (Importance-Performance Analysis). Solid and significant relationships were found among labour conditions, human capital, market and wages, with the relationships between human capital and wages and between human capital and labour conditions being especially relevant. Besides, through IPMA, a series of considerations was made regarding the individual indicators according to their relative importance and performance. Consequently, this paper significantly contributes to the extant literature by analysing the composition of wages in the energy sector, which might allow taking valuable management decisions. Nevertheless, the main limitation of this study lies in the availability of data for Spain and, specifically, for workers in the energy supplying industries.

Social and cultural changes are altering young people's habits. Some surveys measure such behaviors, but are hard to interpret. The objective of this study was to design and validate a homogeneous questionnaire to study habits among adolescents aged 12-14 years.Descriptive and crosssectional study to validate a questionnaire. Items were selected after a bibliographic review and expert assessment. The pre-test was administered to a pilot sample. Reliability was established using Cronbach's alpha coefficient, and construct validity, using a factor analysis. It was administered to a sample of parents of students from 4 secondary schools.Cronbach's coefficient showed values close to 0.7 in 3/4 subscales and an overall value of 0.629, showing accuracy and stability. Factor analysis determined an adequate construct validity, with 4 factors: eating, physical activity, new technologies, and environment. The final questionnaire included 26 items and was administered to 421 participants. Poor habits were observed in 24.8 % and very unhealthy, in 3.4 %. Girls had poorer scores in physical activity (p0.001), and boys, in new technologies, although it was not significant.The final instrument was valid, reliable, and easily administered to assess modifiable behaviors in adolescents. The inclusion of technologies and environment helps to adapt the questionnaire to current changes. The greatest deficit was observed in the technology use and physical activity domains; and girls were more sedentary.Introducción. Los cambios sociales y culturales están alterando los hábitos de los jóvenes. Existen encuestas para medir dichas conductas, pero son de difícil interpretación. El objetivo del estudio fue diseñar y validar un cuestionario homogéneo para estudiar los hábitos en adolescentes de 12- 14 años. Población y métodos. Estudio descriptivo y transversal para validación de un cuestionario. Los ítems fueron seleccionados tras revisión de la literatura y evaluación por expertos. El pretest fue administrado a una muestra piloto. Se determinó la fiabilidad mediante el alfa de Cronbach, y la validez del constructo, mediante un análisis factorial. Se aplicó a una muestra de padres de alumnos de cuatro centros de secundaria. Resultados. El coeficiente de Cronbach mostró valores próximos a 0,7 en tres de las cuatro subescalas y un valor global de 0,629, que demostró precisión y estabilidad. El análisis factorial determinó una validez de constructo adecuada, que mostró 4 factores: alimentación, actividad física, nuevas tecnologías y entorno. El cuestionario final tuvo 26 ítems y se aplicó a 421 participantes. Un 24,8 % presentaron hábitos deficientes, y un 3,4 %, muy poco saludables. Las adolescentes mostraron peores puntuaciones en actividad física (p0,001), y los adolescentes, en nuevas tecnologías, aunque no significativo. Conclusiones. Se obtuvo un instrumento válido, fiable y de fácil aplicación para evaluar conductas modificables en adolescentes. La inclusión de tecnologías y entorno adapta el cuestionario a los cambios actuales. Las dimensiones sobre uso de tecnologías y actividad física fueron las más deficitarias; las adolescentes se mostraron más sedentarias.

We examined in a rat model of Gulf War illness (GWI), the potential of (-)-epicatechin (Epi) to reverse skeletal muscle (SkM) atrophy and dysfunction, decrease mediators of inflammation and normalize metabolic perturbations. Male Wistar rats (n = 15) were provided orally with pyridostigmine bromide (PB) 1.3 mg/kg/day, permethrin (PM) 0.13 mg/kg/day (skin), DEET 40 mg/kg/day (skin) and were physically restrained for 5 min/day for 3 weeks. A one-week period ensued to fully develop the GWI-like profile followed by 2 weeks of either Epi treatment at 1 mg/kg/day by gavage (n = 8) or water (n = 7) for controls. A normal, control group (n = 15) was given vehicles and not restrained. At 6 weeks, animals were subjected to treadmill and limb strength testing followed by euthanasia. SkM and blood sampling was used for histological, biochemical and plasma pro-inflammatory cytokine and metabolomics assessments. GWI animals developed an intoxication profile characterized SkM atrophy and loss of function accompanied by increases in modulators of muscle atrophy, degradation markers and plasma pro-inflammatory cytokine levels. Treatment of GWI animals with Epi yielded either a significant partial or full normalization of the above stated indicators relative to normal controls. Plasma metabolomics revealed that metabolites linked to inflammation and SkM waste pathways were dysregulated in the GWI group whereas Epi, attenuated such changes. In conclusion, in a rat model of GWI, Epi reverses detrimental changes in SkM structure including modulators of atrophy, inflammation and select plasma metabolites yielding improved function.

Consumption of (−)-epicatechin (Epi), a cacao flavanol improves cognition. The aim was to compare the effects of (−)-Epi or its stereoisomer (+)-Epi on mouse frontal cortex-dependent short-term working memory and modulators of neurogenesis. Three-month-old male mice (n = 7 per group) were provided by gavage either water (vehicle; Veh), (−)-Epi, at 1 mg kg−1 or (+)-Epi at 0.1 mg per kg of body weight for 15 days. After treatment, spontaneous alternation was evaluated by Y-maze. Brain frontal cortex was isolated for nitrate/nitrite measurements, Western blotting for nerve growth factor (NGF), microtubule associated protein 2 (MAP2), endothelial and neuronal nitric oxide synthase (eNOS and nNOS) and immunohistochemistry for neuronal specific protein (NeuN), doublecortin (DCX), capillary (CD31) and neurofilaments (NF200). Results demonstrate the stimulatory capacity of (−)-Epi and (+)-Epi on markers of neuronal proliferation as per increases in immunoreactive cells for NeuN (74 and 120% respectively), DCX (70 and 124%) as well as in NGF (34.4, 63.6%) and MAP2 (41.8, 63.8%). Capillary density yielded significant increases with (−)-Epi (∼80%) vs. (+)-Epi (∼160%). CD31 protein levels increased with (−)-Epi (∼70%) and (+)-Epi (∼140%). Effects correlated with nitrate/nitrite stimulation by (–)-Epi and (+)-Epi (110.2, 246.5%) and enhanced eNOS phosphorylation (Ser1177) with (–)-Epi and (+)-Epi (21.4, 41.2%) while nNOS phosphorylation only increased with (+)-Epi (18%). Neurofilament staining was increased in (–)-Epi by 135.6 and 84% with (+)-Epi. NF200 increased with (–)-Epi (116%) vs. (+)-Epi (84.5%). Frontal cortex-dependent short-term spatial working improved with (–)-Epi and (+)-Epi (15, 13%). In conclusion, results suggest that both enantiomers, but more effectively (+)-Epi, upregulate neurogenesis markers likely through stimulation of capillary formation and NO triggering, improvements in memory.

Objectives The main aim of this work was to analyse the potential tumour growth inhibition effects of (−)-epicatechin (EC). Triple-negative breast cancer (TNBC) is an invasive form of cancer characterized by the absence of progesterone receptor, estrogen receptor and human epidermal growth factor receptor 2. Doxorubicin (DOX) is widely used for its anti-tumour activity. EC belongs to the flavanol subfamily and is a candidate molecule for the adjuvant treatment of cancer due to its antiproliferative activities. Methods Evaluation of EC effects and pathways involved in a model of TNBC. Key findings EC inhibited tumour growth as efficiently as DOX (inhibition rates of 74% and 79% for EC and DOX, respectively). The evaluation of adenosine monophosphate-activated protein kinase (AMPK) and Akt phosphorylation and mTOR expression indicates that EC modulates these pathways, resulting in the inhibition of cell proliferation. Additionally, we found an increase in the survival of EC-treated animals compared with control-treated animals. This effect was similar to the effects induced by DOX (survival rates of 44% and 30% for EC and DOX, respectively). Conclusion EC has antiproliferative properties and increases survival in a model of TNBC. These effects may occur through the modulation of deregulated AMPK and Akt/mTOR signalling pathways.

Several studies have evaluated the association between a history of type 2 diabetes (T2D) and risk of breast cancer (BC), with controversial results. However, information regarding the population-attributable risk percent (PAR%) remains scarce. Objective: To estimate the association and the PAR% for BC and T2D, lifestyle and gynecologic factors in women in Mexico City.This case-control study was performed from May-December 2020. Women40 years of age, from Mexico City, with a confirmed diagnosis for BC were included as cases. Controls were women with a BIRADS 1 or 2 mammography or an ultrasound clear of any BC suggestive findings.A total of 134 cases and 134 controls were included. A higher risk for BC was identified among women who did not perform routine physical activity and those who had a history of hormonal contraceptive use5 years. The use of hormone replacement therapy (HRT) was associated with an odds ratio (OR) of 5.0 (p = 0.22) in the first model. After adjustment, HRT was associated with an OR of 2.92 (p=0.492) in the second and an OR of 3.6 (p = 0.753) in the third model. T2D was associated with an OR of 1.04 (p = 0.96) in the first model; an OR of 0.65 (p = 0.65) in the second model and an OR of RMa 0.75(p = 0.79) for the third model.In this case-control study, there was no significant association identified between a T2D diagnosis and BC.diversas investigaciones han evaluado el riesgo de cáncer de mama (CaMa) asociado a diabetes tipo 2 (DT2) con resultados inconsistentes. Pocas investigaciones han evaluado el porcentaje de riesgo atribuible a la población (PAR%).estimar la asociación y el PAR% para CaMa por DT2, así como los factores del estilo de vida y ginecológicos en mujeres de la Ciudad de México.estudio de casos y controles, realizado de mayo a diciembre de 2020. Se incluyeron mujeres40 años, de la Ciudad de México, con diagnóstico confirmado de CaMa. Los controles fueron mujeres con una mastografía BIRADS 1 o 2, o ultrasonido mamario sin datos sugestivos de CaMa.se incluyeron en total 134 casos y 134 controles. Se identificó un mayor riesgo de CaMa en aquellas mujeres que no realizan actividad física y con el uso de anticonceptivos5 años. El uso de terapia de reemplazo hormonal (TRH) se asoció con una razón de momios ajustada (RMa) de 5.0, p = 0.22 en el modelo 1. Conforme se ajustan los modelos, la TRH para el modelo 2 tuvo una RMa de 2.92, p = 0.492 y en el modelo 3 una RMa de 3.6 p = 0.753. La presencia de diabetes se asoció a una RMa de 1.04, p = 0.96 en el modelo 1; RMa de 0.65, p = 0.65 en el modelo 2 y una RMa de 0.75, p = 0.79 en el modelo 3.en este estudio de casos y controles no se encontró asociación entre el diagnóstico de DT2 y CaMa.

In recent years, migratory phenomena have changed the composition of Spanish society. There are many studies that explore the healthy habits of young adolescents, but few focus on young immigrants. The purpose of this study is to examine the causal relationships between certain factors that influence the health of immigrant youth and sports. The sample consisted of 173 students enrolled in secondary education in the city of Cuenca. The structural model confirms the relationship between the latent variables and sports. Specifically, we obtained a positive influence of an active lifestyle (regular physical activity and exercise) and a negative influence for the remaining variables (alcohol consumption, unhealthy foods, self-medication and tobacco consumption in the family). Physical education should promote healthy lifestyles; greater coordination between physical education and other subjects involved in education and the promotion of health are necessary because we consider that this task is not exclusive to physical education.

"Introducción: diversas investigaciones han evaluado el riesgo de cáncer de mama (CaMa) asociado a diabetes tipo 2 (DT2) con resultados inconsistentes. Pocas investigaciones han evaluado el porcentaje de riesgo atribuible a la población (PAR%). Objetivo: estimar la asociación y el PAR% para CaMa por DT2, así como los factores del estilo de vida y ginecológicos en mujeres de la Ciudad de México. Métodos: estudio de casos y controles, realizado de mayo a diciembre de 2020. Se incluyeron mujeres > 40 años, de la Ciudad de México, con diagnóstico confirmado de CaMa. Los controles fueron mujeres con una mastografía BIRADS 1 o 2, o ultrasonido mamario sin datos sugestivos de CaMa. Resultados: se incluyeron en total 134 casos y 134 controles. Se identificó un mayor riesgo de CaMa en aquellas mujeres que no realizan actividad física y con el uso de anticonceptivos > 5 años. El uso de terapia de reemplazo hormonal (TRH) se asoció con una razón de momios ajustada (RMa) de 5.0, p = 0.22 en el modelo 1. Conforme se ajustan los modelos, la TRH para el modelo 2 tuvo una RMa de 2.92, p = 0.492 y en el modelo 3 una RMa de 3.6 p = 0.753. La presencia de diabetes se asoció a una RMa de 1.04, p = 0.96 en el modelo 1; RMa de 0.65, p = 0.65 en el modelo 2 y una RMa de 0.75, p = 0.79 en el modelo 3. Conclusiones: en este estudio de casos y controles no se encontró asociación entre el diagnóstico de DT2 y CaMa."

We aimed to investigate the effects of chronic fluid restriction and hydration with a sweetened beverage (SB) in rats from weaning until adolescence, in a posterior acute kidney injury (AKI) event induced by ischemia-reperfusion (I/R). We followed 5 groups of weaning rats: control group (C); two groups with 22 h/day fluid restriction, a group hydrated for two hours with water (-W) and a group hydrated with SB; one group receiving SB ad libitum all day (+SB); and one group in which water consumption was increased using a gel diet. The rats that reached adolescence were submitted to I/R. Fluid restriction and/or SB hydration induced mild renal alterations that were significantly accentuated in the -SB group and resulted in worse outcomes after I/R-induced AKI that resulted in a catastrophic fall in creatinine clearance and diffuse acute tubular necrosis. In summary, low tap water intakes, as well as SB intake in infancy, prompt kidney worse outcomes in a later event of AKI during adolescence and both insults magnify kidney damage. Studies on hydration habits in children are recommended to disclose the potentially harmful effects that those behavioral patterns might carry to future renal health.

Age-related muscle decline, when associated with obesity, leads to adverse outcomes with increased risks for falling, loss of independence, disability and risk of premature mortality. The aim of this study was to assess the potential beneficial effects of flavonoids in improving the age-/high-fat-diet-induced decrease in physical activity/capacity related to the onset of skeletal muscle decline. The effects of the administration of a cocoa beverage enriched with flavanols or pure (−)-epicatechin for 5 wk in a model of physical activity decline induced by the ingestion of a high-fat diet (60% fat) in middle-age mice were evaluated. The results showed that both products, the cocoa beverage enriched with flavanols and pure (−)-epicatechin, improved physical performance evaluated with the hang-wire, inverted-screen, and weight-lifting tests and dynamometry compared with the performance of the controls. The beverage and (−)-epicatechin increased the follistatin/myostatin ratio and increased the expression of myocyte enhancer factor 2A (MEF2A), suggesting an effect on molecular modulators of growth differentiation. Furthermore, the beverage and (−)-epicatechin decreased the expression of O-type fork-head transcription factor (FOXO1A) and muscle ring finger 1 (MURF1) markers of the skeletal muscle ubiquitin-proteasome degradation pathway.

The skeletal muscle mass reduces 30–60% after spinal cord injury, this is mostly due to protein degradation through ubiquitin–proteasome system. In this work, we propose that the flavanol (−)-epicatechin, due its widespread biological effects on muscle health, can prevent muscle mass decrease after spinal cord injury. Thirty-six female Long Evans rats were randomized into 5 groups: (1) Spinal cord injury 7 days, (2) Spinal cord injury + (−)-epicatechin 7 days, (3) Spinal cord injury 30 days, (4) Spinal cord injury + (−)-epicatechin 30 days and (5) Sham (Only laminectomy). Hind limb perimeter, muscle cross section area, fiber cross section area and ubiquitin–proteasome system protein expression together with total protein ubiquitination were assessed. At 30 days Spinal cord injury group lost 49.52 ± 2.023% of muscle cross section area (−)-epicatechin treated group lost only 24.28 ± 15.45% being a significant difference. Ubiquitin–proteasome markers showed significant changes. FOXO1a increased in spinal cord injury group vs Sham (−)-epicatechin reduced this increase. In spinal cord injury group MAFbx increased significantly vs Sham but decrease in (−)-epicatechin treatment group at 30 days. At 7 and 30 days MuRF1 increased in the spinal cord injury and decreased in the (−)-epicatechin group. The global protein ubiquitination increases after spinal cord injury, epicatechin treatment induce a significant decrease in protein ubiquitination. These results suggest that (−)-epicatechin reduces the muscle waste after spinal cord injury through down regulation of the ubiquitin–proteasome system.

Gulf War illness (GWI) afflicts military personnel who served during the Persian Gulf War and is notable for cognitive deficits, depression, muscle pain, weakness, intolerance to exercise, and fatigue. Suspect causal agents include the chemicals pyridostigmine (PB), permetrim (PM) and N,N-diethyl-m-toluamide (DEET) used as protectants against insects and nerve gases. No pre-clinical studies have explored the effects on skeletal muscle (SkM). Young male rats were provided PB, PM and DEET at equivalent human doses and physical restraint (to induce stress) for 3 weeks followed a 3-week recovery. GWI gastrocnemius weight was ~ 35% lower versus controls, which correlated with decreases in myofiber area, limb strength, and treadmill time/distance. In GWI rats, SkM fiber type relative abundance changed towards slow type I. Muscle wasting pathway proteins were upregulated while those that promote growth decreased as did mitochondrial endpoints and muscle ATP levels. Proteomic analysis of SkM also documented unique alterations in mitochondrial and metabolic pathways. Thus, exposure to GWI chemicals/stress adversely impacts key metabolic pathways leading to muscle atrophy and loss of function. These changes may account for GWI Veterans symptoms.

Head and neck cancer patients are at high risk of anorexia-cachexia syndrome and literature shows that Eicosapentaenoic acid (EPA) could regulate it. We aim to determine the EPA effect on body composition and pro-inflammatory markers in patients with head neck cancer.A randomized single-blind placebo-controlled clinical trial was conducted in patients with head and neck squamous cell cancer who received a polymeric diet with 2 g of EPA or a standard polymeric diet for six weeks before antineoplastic treatment. We assessed body composition by bioelectrical impedance analysis and determined IL-1β, IL-6, TNF-α and IFN-γ, CRP, serum proteins, and blood count at baseline and at the end of the study.32 patients received EPA (2 g/day) and 32 became controls. A decrease in serum levels of IL-1β, IL-6, TNF-α, and IFN-γ was observed in the experimental group, as well as regulation of body weight (-0.3 ± 5.9 vs. -2.1 ± 3.7), lean body mass (-0.2 ± 3.8 vs. -1.3 ± 3.6), body fat mass (0.2 ± 3.5 vs. -1.2 ± 3.8), and quality of life (10 ± 33 vs. 5 ± 34).Supplementing with 2 g/day of EPA to head and neck cancer patient during antineoplastic treatment regulates serum pro-inflammatory cytokines, body weight, lean body mass, and improve quality of life.

The giant Antarctic toothfish Dissostichus mawsoni is one of the most economically valued fishery resources from the Southern Ocean. However, the distinctive biological characteristics and life history of this species, including large body size, long life span, late sexual maturity, and specialization to freezing polar waters, make it particularly vulnerable to overfishing and global climate change. It is therefore of fundamental importance to administer sound management and conservation actions to avoid depletion of this unique species. The crucial issue of whether this species consists of distinct biological units or stocks, which is needed to inform appropriate management and conservation, is currently unresolved. We report here the first RAD-seq analysis of Antarctic toothfish populations, a powerful approach of genome-wide discovery of SNP loci, to assess genetic differentiation between the geographically distant individuals from CCAMLAR Subarea 48.1 locations (South Shetland Islands and West Antarctic Peninsula coasts) and Subarea 88.1 (Ross Sea and McMurdo Sound). We recovered 19,611 SNPs belonging to 13,251 widely shared loci. Comprehensive population structure analyses unambiguously indicated absence of any significant level of population differentiation, thus the null hypothesis of panmixia cannot be rejected. Together with currently known life history traits of D. mawsoni, we suggest this striking lack of genomic differentiation likely reflects high degrees of contemporary and/or historical gene flow rather than shared spawning grounds. Complementary analyses, particularly otolith microchemistry, would contribute useful inference of natal origins.

The production of nitric oxide (NO) by nitric oxide synthases (NOS) depends on the bioavailability of L-arginine as NOS competes with arginase for this common substrate. As arginase activity increases, less NO is produced and adverse cardiovascular consequences can emerge. (-)-Epicatechin (EPI), the most abundant flavonoid in cacao, has been reported to stimulate endothelial and neuronal NOS expression and function leading to enhanced vascular function and cardioprotective effects. However, little is known about the effects of EPI on myocardial arginase activity. The aim of the present study was to determine if EPI is able to interact and modulate myocardial arginase and NOS expression and activity. For this purpose, in silico modeling, in vitro activity assays and a rat model of ischemia/reperfusion injury were used. In silico and in vitro results demonstrate that EPI can interact with arginase and significantly decrease its activity. In vivo, 10 days of EPI pretreatment reduces ischemic myocardium arginase expression while increasing NOS expression and phosphorylation levels. Altogether, these results may partially account for the cardioprotective effects of EPI.

The consumption of cocoa products rich in (−)-epicatechin is associated with reduced cardiovascular risk and improved vascular function. However, little is known about (−)-epicatechin’s effects on aged endothelium. In order to characterize the health restoring effects of (−)-epicatechin on aged endothelium and identify the underlying mechanisms, we utilized high passage number (i.e. aged) bovine coronary artery endothelial cells and aortas of 3 and 18 month old rats. We evaluated cell senescence (β-galactosidase), nitric oxide (NO) production through the endothelial nitric oxide synthase pathway, mitochondria related endpoints, citrate synthase activity and vascular relaxation. Cells were treated with water or (−)-epicate-chin (1 μM) for 48 h and rats orally with either water or (−)-epicatechin (1 mg kg(−1) day(−1)) for 15 days. Senescence associated β-galactosidase levels doubled in aged cells while those treated with (−)-epicate-chin only evidenced an ~40% increase. NO levels in cells decreased by ~33% with aging and (−)-epicate-chin normalized them. Endothelial nitric oxide synthase phosphorylation levels paralleled these results. Aging increased total protein and synthase acetylation levels and (−)-epicatechin partially restored them to those of young cells by stimulating sirtuin-1 binding to the synthase. Phosphorylated sirtuin-1, mitofilin, oxidative phosphorylation complexes and transcriptional factor for mitochondria were reduced by ~40% with aging and were restored by (−)-epicatechin. (−)-Epicatechin enhanced acetylcholine induced aged aorta vasodilation and stimulated NO levels while reducing blood pressure. In conclusion, (−)-epicatechin reverses endothelial cell aging and restores key control elements of vascular function. These actions may partly explain the epidemiological evidence for the beneficial effects of cocoa consumption on the incidence of cardiac and vascular diseases.

We reported that (-)-epicatechin can stimulate mitochondria biogenesis and improve metabolism. However, preliminary studies indicate that the (+) stereoisomer form may be more potent. We evaluated in a preliminary manner, the pharmacokinetics (PK) and initial safety analysis of (+)-epicatechin ((+)-Epi) in healthy and pre-diabetic subjects. Using a mouse model of diet-induced obesity and insulin resistance, we also evaluated the metabolic effects of (+)-Epi vs. (+)-catechin (Cat) to determine class effects. In the Phase I PK study, subjects were provided a single incremental oral dose of (+)-Epi (10, 30 or 100 mg). For the PD study, subjects were provided a single 30 mg dose per day for 7 days. Blood samples were collected and safety measures were performed. Incremental doses of (+)-Epi increase the half-life of blood metabolites from 1.2-4.9 h. The compound was well tolerated and no adverse effects were reported. Seven day dosing of pre-diabetic subjects led to tendencies for reductions in circulating levels of tumor necrosis factor-α and monocyte chemoattractant protein-1, which returned to baseline by 7 days after treatment. In animals, 2 weeks of oral dosing (0.003, 0.01, 0.03, 0.1 and 0.3 mg kg-1 day-1) dose dependently improved metabolism-related endpoints (weight gain, glucose, cholesterol, triglyceride, with thresholds as low as 0.01 mg kg-1 day-1). Cat yielded no effects at 0.1 mg kg-1 day-1. Results indicate that (+)-Epi evidences a favorable PK and safety profile. Using a pre-clinical model, the compound positively modulates metabolism, which may link to mitochondrial effects. Effects are not due to general antioxidant actions, as Cat yielded no effects.

Introduction Multiple components of the dystrophin-associated protein complex (DAPC) are expressed in numerous tissues including the brain. Members of the DAPC and dysbindin are abnormally expressed in the brain of Duchenne Muscular Dystrophy (DMD) patients, which has been associated with cognitive impairments. However, little is known about the expression pattern of individual members of the DAPC in animal models of DMD and their relationship with dysbindin. Methods Ten mdx mice were randomly allocated into a control and intervention group [(−)-epicatechin (Epi) 1 mg/kg/day for four weeks] and results compared to a wild-type mice. After sacrifice, brain pre-frontal cortices were collected for Western blotting and immunoprecipitation assays, and sagittal sections processed for immunohistochemistry. Results Epi promotes a partial recovery of DAPC members [α1-Syntrophin, sarcoglycans (SG), dystrophin 71 (Dp71)], dysbindin, and utrophin protein levels. Epi also appears to restore the association of DAPC between dysbindin, and utrophin with Dp71 and e-SG. Co-immunostaining evidence increased protein levels of dysbindin, dystrophin, and e-SG and their colocalization. Conclusions Altogether, results suggest that Epi is capable of restoring pre-frontal cortex DAPC and dysbindin levels of mdx mice towards that of healthy brains. The functional implications of such studies warrant further investigation.

The cacao pod husk (CPH) is the primary waste byproduct of the chocolate industry. One ton of cacao beans generate approximately ten times that amount of fresh CPH. The husk is rich in phenolic compounds. The aim of this study is to determine the effects of microwaves, hot air and freeze-drying on the phenolic compound content, antioxidant capacity, enzyme activity and microstructure of CPH. The results showed that fresh CPH contain 323.7 ± 26.5 mg gallic acid equivalents (GAE)/100 g dry mass (d.m.) of total phenolic compounds, and dehydration had a positive effect on the CPH phenolic content and antioxidant capacity. Catechin, quercetin, and (−)-epicatechin and gallic, coumaric and protocatechuic acids were identified in fresh and dried CPH. Drying the material using a microwave and freeze-drying preserved the husk microstructure. The results showed that microwave drying was a better drying method than hot air and as good as freeze-drying. Industrial relevance The food industry is currently searching for new sources of natural antioxidants. The cacao pod husk (CPH) is a good source of phenolic compounds that can function as antioxidants and could be used as ingredients in functional foods. However, due to their high water content, fresh CPH are perishable products, and for this reason, the first step would be dry the pod husk to improve its shelf life. The findings of this study showed that the use of microwaves to dry CPH may release polyphenols, thereby enhancing the antioxidant capacity and decreasing the polyphenol oxidase (PPO) activity while maintaining the microstructure. Thus, microwave drying could be of interest as a method for enhancing the extractability of phenolic compounds from CPH.

Treating addiction and withdrawal syndrome remains a research area of great interest since it is a persistent public health problem. Opiates account for the majority of addiction and withdrawal problems (at least 56% of reported people who consume drugs). Development of opiate addiction results mainly from activation of the mu-opioid receptor (MOR), and this entity is identified as a major target for the treatment of addiction and withdrawal syndrome. Based on our in silico analysis and calculations of binding energy, conformations adopted by catechins and contacts reached by ligand conformations could explain why opiates are capable of activating both β-arrestin and G-protein biased pathways, and therefore help to explain the phenomenon of tolerance, we can conclude that the family of catechins explored seem to be excellent modulators of the MOR.

The development of organ fibrosis has garnered rising attention as multiple diseases of increasing and/or high prevalence appear to progress to the chronic stage. Such is the case for heart, kidney, liver, and lung where diseases such as diabetes, idiopathic/autoimmune disorders, and nonalcoholic liver disease appear to notably drive the development of fibrosis. Noteworthy is that the severity of these pathologies is characteristically compounded by aging. For these reasons, research groups and drug companies have identified fibrosis as a therapeutic target for which currently, there are essentially no effective options. Although a limited body of published studies are available, most literature indicates that in multiple organs, premenopausal women are protected from developing severe forms of fibrosis suggesting an important role for sex hormones in mitigating this process. Investigators have implemented relevant animal models of organ disease linked to fibrosis supporting in general, these observations. In vitro studies and transgenic animals models have also been used in an attempt to understand the role that sex hormones and related receptors play in the development of fibrosis. However, in the setting of chronic disease in some organs such as the heart older (postmenopausal) women within a few years can quickly approach men in disease severity and develop significant degrees of fibrosis. This review summarizes the current body of relevant literature and highlights the imperative need for a major focus to be placed on understanding the manner in which sex and the presence or absence of related hormones modulates cell phenotypes so as to allow for fibrosis to develop.

Deep vein thrombosis (DVT) is a common complication during postoperative convalescence characterized by hypercoagulability, vascular endothelium damage and blood stasis. It increases noticeably in peri/postoperative phases of surgery procedures. Pulmonary embolism secondary to iliofemoral DVT is a frequent cause of death. Adult patients scheduled for plastic and reconstructive surgery (PRSx) with moderate to high thrombogenic risk were selected. We evaluated the efficacy and safety of bemiparin compared to enoxaparin as chemoprophylaxis for DVT. Following balanced general anesthesia techniques, patients were randomly assigned for subcutaneous enoxaparin 40 IU (Group-E) or bemiparin 3500 IU (Group-B) q24h starting 6 h after procedure conclusion for at least 10 days. All patients were evaluated for DVT through Doppler ultrasound mapping of the lower limbs. Seventy-eight patients were evaluated, mostly women (83%), physical status ASA II (59%), ASA III (10%); Caprini’s thrombogenic risk score 3–4 (moderate) 58%, 5–6 (high) 29%, > 6 (too high) 13%; demographics, clinical variables and scores were similar between groups. Median drainage time in breast surgery was 4 days in both groups (p = 0.238). In the case of abdominal surgery, median was 14 days in Group-E versus 13 days in Group-B (p = 0.059). No DVT was detected in either group. DVT was prevented with bemiparin, without significant bleeding increase nor adverse events; moreover, the cost of bemiparin is lower than enoxaparin. Bemiparin can be considered as alternative drug for DVT chemoprophylaxis in PRSx procedures. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.

Introduction We conducted an open‐label study to examine the effects of the flavonoid (−)‐epicatechin in seven ambulatory adult patients with Becker muscular dystrophy (BMD). Methods Seven participants received (−)‐epicatechin 50 mg twice per day for 8 weeks. Pre‐ and postprocedures included biceps brachii biopsy to assess muscle structure and growth‐relevant endpoints by western blotting, mitochondria volume measurement, and cristae abundance by electron microscopy, graded exercise testing, and muscle strength and function tests. Results Western blotting showed significantly increased levels of enzymes modulating cellular bioenergetics (liver kinase B1 and 5′‐adenosine monophosphate–activated protein kinase). Peroxisome proliferator‐activated receptor gamma coactivator‐1alpha, a transcriptional coactivator of genes involved in mitochondrial biogenesis and cristae‐associated mitofilin levels, increased as did cristae abundance. Muscle and plasma follistatin increased significantly while myostatin decreased. Markers of skeletal muscle regeneration myogenin, myogenic regulatory factor‐5, myoblast determination protein 1, myocyte enhancer factor‐2, and structure‐associated proteins, including dysferlin, utrophin, and intracellular creatine kinase, also increased. Exercise testing demonstrated decreased heart rate, maximal oxygen consumption per kilogram, and plasma lactate levels at defined workloads. Tissue saturation index improved in resting and postexercise states. Discussion (−)‐Epicatechin, an exercise mimetic, appears to have short‐term positive effects on tissue biomarkers indicative of mitochondrial biogenesis and muscle regeneration, and produced improvements in graded exercise testing parameters in patients with BMD.

To characterize the distribution of the foveal avascular zone circularity and its correlation with parafoveal vessel density, in subjects with and without diabetes.Observational, descriptive, cross-sectional, and prospective study; subjects without diabetes (Group 1), with diabetes without retinopathy (Group 2), or with diabetic retinopathy (Group 3) were included. Means of foveal avascular zone circularity and parafoveal vessel density were compared between groups (Kruskal-Wallis) and their correlation was calculated with Spearman's Rho test.Seventy-seven eyes; central vessel density mean was higher in Group 1 than in Group 2 and higher in Group 2 than in Group 3; inner and complete vessel density means were also higher in Group 2 than in Group 3. The mean of the foveal avascular zone circularity did not differ between groups, and in Group 3 it had a positive correlation with central (0.45), inner (0.56), and complete (0.53) vessel densities.Circularity does not differ between subjects with diabetes, with and without retinopathy, and has only a low correlation with parafoveal vessel density in people with diabetic retinopathy, which does not allow anticipating a reduction of vessel density in this disease.Caracterizar la distribución de la circularidad de la zona avascular foveal y su correlación con la densidad vascular perifoveal, en sujetos con y sin diabetes.Estudio observacional, descriptivo, transversal y prospectivo; se incluyeron sujetos sin diabetes (Grupo 1), con diabetes sin retinopatía (Grupo 2) y con retinopatía diabética (Grupo 3). Los promedios de circularidad de la zona avascular foveal y de la densidad vascular parafoveal se compararon entre grupos (Kruskal-Wallis), y se calculó su correlación mediante la prueba Rho de Spearman.Se estudiaron 77 ojos. El promedio de la densidad vascular central fue mayor en el Grupo 1 que en el Grupo 2, y mayor en el Grupo 2 que en el Grupo 3. Los promedios de la densidad vascular interna y completa también fueron mayores en el Grupo 2 que en el Grupo 3. El promedio de la circularidad de la zona avascular foveal no difirió entre grupos, y en el Grupo 3 tuvo una correlación positiva con la densidad vascular central (0.45), interna (0.56) y completa (0.53).La circularidad no difiere entre sujetos con y sin diabetes, con y sin retinopatía, y solo tiene una baja correlación con la densidad vascular parafoveal en sujetos con retinopatía diabética, lo cual no permite anticipar una reducción de la densidad vascular en esta enfermedad.

The effects of testosterone on cardiovascular homeostasis are still not well understood. The objective of this work was to evaluate the effects of testosterone in the absence or presence of inhibition of Aromatase (4-hydroxyandrostenedione) and/or 5α reductase (Finasteride) enzymatic activities on the myocardial remodeling 30 days after ischemia/reperfusion (I/R) injury in gonadectomized rats. Results showed that testosterone administration to ORX rats resulted in decreased myocardial damaged area, inflammatory infiltrates and reduced MMP-3 and 13 expressions. Interestingly, Finasteride administration resulted in a greater decrease in scar tissue, inflammatory infiltrates, along with a significant decrease in MMP-3 and 13 expressions. In contrast, 4-hydroxyandrostenedione administrations increased all parameters. Our results suggest that testosterone does not have a direct effect since simultaneous inhibition of aromatase and 5α-reductase did not induce significant changes in I/R induced myocardial injury., Summary: Coronary ischemia/reperfusion-induced injury in gonadectomyzed male rats is decreased by testosterone, protection is increased by blocking its 5α-reduction and blocked by inhibition of its aromatization.

Background The age-related decline in mass, strength, and performance of skeletal muscle is associated with loss of independence, falls risk, disability, institutionalization, and death. Methods To determine whether a cocoa supplement enriched in flavonoids can improve plasma markers of oxidative stress and inflammation, physical performance and frailty in middle-aged and older subjects, we conducted a two-phase, randomized, double-blind, clinical trial. The initial study included 60 subjects (55- to 70-year-old) allocated into placebo (P), highly alkalinized (no-flavonoid; NF), or flavonoid-rich natural cocoa (F) beverage groups. The follow-up study included 74 older subjects (65- to 90-year-old) randomly distributed into NF or F groups. Subjects were instructed to consume the beverages once/day for up to 12-weeks. A comprehensive (aging relevant) set of end points were assessed, which included mean change in blood plasma metabolic and oxidative stress indicators, in physical performance tests and quality of life (QoL). Results In the initial study, the F group showed improved glycemia, triglyceridemia, High-density lipoprotein cholesterol, Low-density lipoprotein cholesterol, triglyceridemia/HDL index, and oxidative markers. Performance on the Up and Go test, skeletal muscle index, and quality of life also improved. In the follow-up study, F treatment was associated with significant improvements in metabolic, oxidative stress, and inflammatory endpoints and positive effects on physical performance, frailty indicators, and quality of life (F vs. NF group). Conclusions Regular flavonoids consumption positively affects blood oxidative stress and inflammation end points, cardiometabolic risk markers, physical performance, and quality of life. The sum of such effects may help to mitigate the extent of frailty development in the elderly people. Trial Registration NCT03585868

Aims To evaluate the effects of (-)-epicatechin (Epi) in the progression of kidney damage. Material and methods We assessed the effects of Epi [0.01–20 mg/kg of body weight/day] during 14 days, in a 5/6 nephrectomy model in mice. Key findings Nephrectomy-induced systolic arterial hypertension was significantly reduced in a dose dependent manner with Epi treatment. Increased serum creatinine and urea were reduced almost to normal values. The concentration of tetrahydrobiopterin (BH4), used as subrogate of endothelial dysfunction, decreased in nephrectomyzed animals, Epi treatment increased BH4 levels almost reaching normal values. The expression of angiotensin II receptor (AT1-R) and NADPH oxidase-4 (NOX-4) and 3-nitrotyrosine levels increased with nephrectomy and were reduced with Epi treatment. Renal tissue morphology in the remaining tissue was conserved with Epi treatment in a dose dependent manner. Significance. Chronic kidney disease (CKD) is an independent cardiovascular risk factor associated with a mortality rate 10 to 20 times higher than that of the general population. High blood pressure, endothelial dysfunction and oxidative stress are important factors determining kidney damage progression. Findings of this study indicate that Epi is able to counteract the deleterious effects of subtotal nephrectomy and the structural and functional changes in the remnant kidney tissue, decreasing the progression of CKD. These results warrant the possibility of implement clinical trials to limit the progression of CKD in humans.

(-)-Epicatechin increases indicators associated with mitochondrial biogenesis in endothelial cells and myocardium. We investigated endothelial nitric oxide synthase involvement on (-)-epicatechin-induced increases in indicators associated with mitochondrial biogenesis in human coronary artery endothelial cells cultured in normal-glucose and high-glucose media, as well as to restore indicators of cardiac mitochondria from the effects of simulated diabetes. Here, we demonstrate the role of endothelial nitric oxide synthase on (-)-epicatechin-induced increases in mitochondrial proteins, transcription factors and sirtuin 1 under normal-glucose conditions. In simulated diabetes endothelial nitric oxide synthase function, mitochondrial function–associated and biogenesis-associated indicators were adversely impacted by high glucose, effects that were reverted by (-)-epicatechin. As an animal model of type 2 diabetes, 2-month old C57BL/6 mice were fed a high-fat diet for 16 weeks. Fasting and fed blood glucose levels were increased and NO plasma levels decreased. High-fat-diet-fed mice myocardium revealed endothelial nitric oxide synthase dysfunction, reduced mitochondrial activity and markers of mitochondrial biogenesis. The administration of 1 mg/kg (-)-epicatechin for 15 days by oral gavage shifted these endpoints towards control mice values. Results suggest that endothelial nitric oxide synthase mediates (-)-epicatechin-induced increases of indicators associated with mitochondrial biogenesis in endothelial cells. (-)-Epicatechin also counteracts the negative effects that high glucose or simulated type 2 diabetes has on endothelial nitric oxide synthase function.

Endothelial dysfunction (EnD) occurs with aging and endothelial nitric oxide (NO) production by NO synthase (NOS) can be impaired. Low NO levels have been linked to increased arginase (Ar) activity as Ar competes with NOS for L-arginine. The inhibition of Ar activity can reverse EnD and (−)-epicatechin (Epi) inhibits myocardial Ar activity. In this study, through in silico modeling we demonstrate that Epi interacts with Ar similarly to its inhibitor Norvaline (Norv). Using in vitro and in vivo models of aging, we examined Epi and Norv-inhibition of Ar activity and its endothelium-protective effects. Bovine coronary artery endothelial cells (BCAEC) were treated with Norv (10 μM), Epi (1 μM) or the combination (Epi + Norv) for 48 h. Ar activity increased in aged BCAEC, with decreased NO generation. Treatment decreased Ar activity to levels seen in young cells. Epi and Epi + Norv decreased nitrosylated Ar levels by ∼25% in aged cells with lower oxidative stress (∼25%) (dihydroethidium) levels. In aged cells, Epi and Epi + Norv restored the eNOS monomer/dimer ratio, protein expression levels and NO production to those of young cells. Furthermore, using 18 month old rats 15 days of treatment with either Epi (1 mg/kg), Norv (10 mg/kg) or combo, decreased hypertension and improved aorta vasorelaxation to acetylcholine, blood NO levels and tetra/dihydribiopterin ratios in cultured rat aortic endothelial cells. In conclusion, results provide evidence that inhibiting Ar with Epi reverses aged-related loss of eNOS function and improves vascular function through the modulation of Ar and eNOS protein levels and activity., Highlights • In aged endothelium, arginase (Ar) activity is increased, leading to a reduction in NO production. • (−)-Epicatechin (Epi) inhibits Ar activity and modulates L-arginine availability. • Inhibition of Ar with Epi reverses aged-related loss of eNOS function. • Epi and Epi plus Norvaline improves endothelial and vascular function in aged endothelium.

Background: The impressive adaptive radiation of notothenioid fishes in Antarctic waters is generally thought to have been facilitated by an evolutionary key innovation, antifreeze glycoproteins, permitting the rapid evolution of more than 120 species subsequent to the Antarctic glaciation. By way of contrast, the second-most species-rich notothenioid genus, Patagonotothen, which is nested within the Antarctic clade of Notothenioidei, is almost exclusively found in the non-Antarctic waters of Patagonia. While the drivers of the diversification of Patagonotothen are currently unknown, they are unlikely to be related to antifreeze glycoproteins, given that water temperatures in Patagonia are well above freezing point. Here we performed a phylogenetic analysis based on genome-wide single nucleotide polymorphisms (SNPs) derived from restriction site-associated DNA sequencing (RADseq) in a total of twelve Patagonotothen species. Results: We present a well-supported, time-calibrated phylogenetic hypothesis including closely and distantly related outgroups, confirming the monophyly of the genus Patagonotothen with an origin approximately 3 million years ago and the paraphyly of both the sister genus Lepidonotothen and the family Notothenidae. Our phylogenomic and population genetic analyses highlight a previously unrecognized linage and provide evidence for shared genetic variation between some closely related species. We also provide a mitochondrial phylogeny showing mitonuclear discordance. Conclusions: Based on a combination of phylogenomic and population genomic approaches, we provide evidence for the existence of a new, potentially cryptic, Patagonotothen species, and demonstrate that genetic boundaries between some closely related species are diffuse, likely due to recent introgression and/or incomplete linage sorting. The detected mitonuclear discordance highlights the limitations of relying on a single locus for species barcoding. In addition, our time-calibrated phylogenetic hypothesis shows that the early burst of diversification roughly coincides with the onset of the intensification of Quaternary glacial cycles and that the rate of species accumulation may have been stepwise rather than constant. Our phylogenetic framework not only advances our understanding of the origin of a high-latitude marine radiation, but also provides the basis for the study of the ecology and life history of the genus Patagonotothen, as well as for their conservation and commercial management. Fil: Ceballos, Santiago Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Austral de Investigaciones Científicas; Argentina. Universidad Nacional de Tierra del Fuego; Argentina Fil: Roesti, Marius. University of Basel,; Suiza. University of British Columbia; Canadá Fil: Matschiner, Michael. University of Basel,; Suiza. University of Oslo; Noruega Fil: Fernandez, Daniel Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Austral de Investigaciones Científicas; Argentina. Universidad Nacional de Tierra del Fuego; Argentina Fil: Damerau, Malte. Johann Heinrich von Thünen-Institut ; Alemania Fil: Hanel, Reinhold. Johann Heinrich von Thünen-Institut ; Alemania Fil: Salzburger, Walter. University of Basel,; Suiza

(-)-Epicatechin (Epi) has been demonstrated to activate pathways involved in GPER-stimulated nitric oxide (NO) production via endothelial NO synthase, known as the eNOS/NO pathway. Previous studies combining synthesis of four Epi derivatives demonstrated that Epi and Epi-prop, Epi-4-prop and Epi-5-prop were able to bind GPER by acting as GPER agonists, whereas docking studies allowed observation of structural details of the binding of these derivatives at the GPER binding site. However, due to the nature of past studies, the theoretical methods employed did not allow observation of structural and energetic details linked to ligand binding at the GPER binding site. In this contribution, we explore the structural and energetic changes coupling the binding of Epi and its four derivatives to GPER. To this end, MD simulations on the microsecond scale (1 μs) with an MMGBSA approach were used for each GPER-ligand complex. Energetic analysis demonstrated that incorporation of several aliphatic chains to Epi contributed to increasing the affinity towards the GPER binding site, thus helping to explain the experimental evidence. Structural analysis demonstrated that Epi, Epi-4-prop and Epi-5-prop share more similar interactions at GPER binding sites with similar conformational behavior than with Epi-prop and Epi-Ms. However, Epi-prop had additional residues that could explain its different but related biological effects.