AIMSNorepinephrine (NE) is a known regulator of adipose tissue (AT) metabolism, angiogenesis, vasoconstriction and fibrosis. This may be through autocrine/paracrine effects on local resistance vessel ...function and morphology. The aims of this study were to investigate, in human subcutaneous and omental adipose tissue (SAT and OAT): NE synthesis, angiogenesis, NE-mediated arteriolar vasoconstriction, the induction of collagen gene expression and its deposition in non-diabetic versus diabetic obese subjects. MATERIALS AND METHODSSAT and OAT from obese patients were used to investigate tissue NE content, tyrosine hydroxylase (TH) density, angiogenesis including capillary density, angiogenic capacity and angiogenic gene expression, NE-mediated arteriolar vasoconstriction and collagen deposition. KEY FINDINGSIn the non-diabetic group, NE concentration, TH immunoreactivity, angiogenesis and maximal vasoconstriction were significantly higher in OAT compared to SAT (p < 0.05). However, arterioles from OAT showed lower NE sensitivity compared to SAT (10-8 M to 10-7.5 M, p < 0.05). A depot-specific difference in collagen deposition was also observed, being greater in OAT than SAT. In the diabetic group, no significant depot-specific differences were seen in NE synthesis, angiogenesis, vasoconstriction or collagen deposition. SAT arterioles showed significantly lower sensitivity to NE (10-8 M to 10-7.5 M, p < 0.05) compared to the non-diabetic group. SIGNIFICANCESAT depot in non-diabetic obese patients exhibited relatively low NE synthesis, angiogenesis, tissue fibrosis and high vasoreactivity, due to preserved NE sensitivity. The local NE synthesis in OAT and diabetes desensitizes NE-induced vasoconstriction, and may also explain the greater tissue angiogenesis and fibrosis in these depots.
Ecdysteroids are of interest as potential sport performance enhancers, due to their anabolic effects. The current study aimed to analyze levels of the most abundant ecdysteroid, ecdysterone ...(20‐hydroxyecdysone, 20‐OHE) in easily available dietary supplements, and, outline an analytical strategy for its detection, and that, of its metabolites, (1) following administration of pure 20‐OHE to uPA(+/+)‐SCID mice with humanized liver, (2) in a human volunteer after ingestion of two supplements, one with a relatively low, and the other a high, concentration of 20‐OHE, and, (3) to estimate the prevalence of use of 20‐OHE in elite athletes (n = 1000). Of the 16 supplements tested, only five showed detectable levels of 20‐OHE, with concentrations ranging from undetectable up to 2.3 mg per capsule. Urine of uPA(+/+)‐SCID urine showed the presence of 20‐OHE and its metabolite, 14 deoxy ecdysterone, within 24 hours (hr) of ingestion. In humans, both the parent and the metabolite were detectable within 2 to 5 hr of ingestion, with the metabolite being detectable for longer than the parent. After ingestion of a low dose supplement, the parent and metabolite were detectable for 70 and 48 hr, while following the higher dose it was 96 and 48 hr, respectively. Analysis of urines from athletes (n = 1000) confirmed four positives for 20‐OHE, suggesting a prevalence of use of 0.4%. Prevalence of its use by elite athletes was relatively low, however, this needs to be confirmed in other populations, and with other related ecdysteroids.
Ecdysteroids have recently been included in the WADA Monitoring program due to their anabolic effects. Concentrations of 20 OHE in dietary supplements are variable. Urinary 20‐OHE and 14‐deoxy‐20‐OHE were detected in uPA(+/+)−SCID mice within 24 hours (hr) of ingestion. In humans, 20‐OHE and its metabolite were detectable for 70 and 48 hr (2 caps of 0.0088 mg) and for 96 and 48 hr (two capsules of 2.3 mg). In elite athletes there was a prevalence of use of 0.4%.
Loss of function mutations in store‐operated Ca2+ entry (SOCE) are associated with severe paediatric disorders in humans, including combined immunodeficiency, anaemia, thrombocytopenia, anhidrosis ...and muscle hypotonia. Given its central role in immune cell activation, SOCE has been a therapeutic target for autoimmune and inflammatory diseases. Treatment for such chronic diseases would require prolonged SOCE inhibition. It is, however, unclear whether chronic SOCE inhibition is viable therapeutically. Here we address this issue using a novel genetic mouse model (SOCE hypomorph) with deficient SOCE, nuclear factor of activated T cells activation, and T cell cytokine production. SOCE hypomorph mice develop and reproduce normally and do not display muscle weakness or overt anhidrosis. They do, however, develop cardiovascular complications, including hypertension and tachycardia, which we show are due to increased sympathetic autonomic nervous system activity and not cardiac or vascular smooth muscle autonomous defects. These results assert that chronic SOCE inhibition is viable therapeutically if the cardiovascular complications can be managed effectively clinically. They further establish the SOCE hypomorph line as a genetic model to define the therapeutic window of SOCE inhibition and dissect toxicities associated with chronic SOCE inhibition in a tissue‐specific fashion.
Key points
A floxed stromal interaction molecule 1 (STIM1) hypomorph mouse model was generated with significant reduction in Ca2+ influx through store‐operated Ca2+ entry (SOCE), resulting in defective nuclear translocation of nuclear factor of activated T cells, cytokine production and inflammatory response. The hypomorph mice are viable and fertile, with no overt defects.
Decreased SOCE in the hypomorph mice is due to poor translocation of the mutant STIM1 to endoplasmic reticulum–plasma membrane contact sites resulting in fewer STIM1 puncta.
Hypomorph mice have similar susceptibility to controls to develop diabetes but exhibit tachycardia and hypertension. The hypertension is not due to increased vascular smooth muscle contractility or vascular remodelling.
The tachycardia is not due to heart‐specific defects but rather seems to be due to increased circulating catecholamines in the hypomorph.
Therefore, long term SOCE inhibition is viable if the cardiovascular defects can be managed clinically.
figure legend Summary of the functional and signalling changes in the stromal interaction molecule 1 hypomorph mouse model with associated toxicities.
Antidoping testing for recombinant human erythropoietin (EPO) is routinely performed by gel electrophoresis followed by western blot analysis with primary and secondary antibodies. The two antibody ...steps add more than 24 h to the testing time of a purified sample. The aim of this study was to test the concept of using directly horseradish‐peroxidase (HRP)‐conjugated anti‐EPO primary antibody, without the need for a secondary antibody, to reduce the analysis time and eliminate non‐specific cross‐reactivity with secondary antibodies. An in‐house, periodate coupling (R&D systems, clone AE7A5) and three commercially available anti‐human EPO‐HRP conjugates from Genetex, Novus Biologicals and Santa Cruz were evaluated for specificity and sensitivity, using recombinant human EPO standards, negative human urine samples and urine samples from an EPO excretion study. The in‐house anti‐EPO‐HRP conjugate was performed as well as the current two‐step application of unconjugated primary and secondary antibodies used in routine analysis, with comparable specificity and sensitivity. The analysis time was markedly reduced for purified samples from 25 h with the routine method down to 7 h with the in‐house HRP conjugate. Of the three commercially available conjugates tested, only the Santa Cruz anti‐EPO‐HRP conjugate showed comparable specificity but had lower sensitivity to both the in‐house and the antibody combination currently applied routinely. The other two commercially available conjugates (Genetex and Novus Biologicals) did not show any visible bands with the EPO standards. The results clearly demonstrate the potential utility of a directly HRP‐conjugated anti‐EPO antibody to reduce analysis time for EPO in doping control.
Detection of EPO is usually performed by western blot with primary and secondary antibodies. The method evaluated here tested successfully the concept of using a directly horseradish‐peroxidase (HRP)‐conjugated anti‐EPO primary antibody and reduced the EPO primary antibody and the necessary analysis time from 25 to 7 h.
OBJECTIVE—To investigate the hypothesis that release of adipokines by epicardial adipose tissue (EAT) is dysregulated in obesity and/or coronary artery disease (CAD), along with the previously ...documented expansion of the tissue, and that these molecules induce pathophysiological changes in human monocytes and coronary artery endothelial cells.
METHODS AND RESULTS—In white nondiabetic patients with CAD (n=62) or without CAD (control group) (n=32), subdivided by body mass index of ≤27 and >27, 13 cytokines were identified by protein array analysis as EAT products. Interleukin 6, interleukin 8, monocyte chemoattractant protein 1, plasminogen activator inhibitor 1, growth-related oncogene-α, and macrophage migration inhibitory factor were the most abundant. Adiponectin release was suppressed in patients with obesity and CAD, and regulated on activation T-cell and secreted (RANTES) was induced in patients with CAD. EAT-conditioned media induced migration of monocytic tryptophan hydroxylase 1 (THP-1) cells, an effect exacerbated in those with CAD. Moreover, conditioned media from patients with CAD and body mass index of >27 increased the adhesion of THP-1 cells to human coronary artery endothelial cells by 15.1% (P=0.002) and expression of intercellular adhesion molecule 1 by 2.8-fold (P=0.002). This effect was reversed by recombinant adiponectin.
CONCLUSION—EAT products are altered in both obesity and CAD and induce atherogenic changes in relevant target cells.
Abstract
Objective: Inflammation has been implicated in the pathogenesis of postoperative atrial fibrillation (AF). Adipose tissue secretes both pro-inflammatory cytokines such as interleukin-6 ...(IL-6) and anti-inflammatory mediators such as adiponectin. We set out to examine the association of adiponectin and IL-6, both circulating and locally produced by the epicardial adipose tissue, with AF development after cardiac surgery. Methods: A total of 90 consecutive patients undergoing cardiac surgery were evaluated. Blood samples were collected before induction of anaesthesia. Epicardial fat was obtained upon commencement of cardiopulmonary bypass. IL-6 and adiponectin levels were determined in serum and supernatant of epicardial adipose tissue organ cultures with two-site enzyme-linked immunosorbent assay (ELISA). Heart rhythm was assessed with continuous tele-monitoring for 72 h postoperatively, and with 6-hourly clinical examinations and daily electrocardiograms (ECGs) thereafter. Results: A total of 36 patients developed postoperative AF (40%). Baseline-serum IL-6 and adiponectin were not associated with AF (p = 0.86 and 0.95, respectively). Epicardial adipose tissue IL-6 levels did not correlate with the development of the arrhythmia either (p = 0.37). However, epicardial adiponectin release was lower in patients who developed AF than in those who remained in sinus rhythm (76 (interquartile range (IQR) 35-98) vs 53 ((IQR) 35-69) ng h−1 g−1 of tissue cultured, p = 0.066). Following linear regression, the association of epicardial adiponectin with AF almost reached statistical significance (p = 0.066). Multivariate logistic regression analysis of identified risk factors for AF, with the inclusion of epicardial adiponectin as an independent variable, revealed increased age (odds ratio (OR) 1.09, 95% confidence interval (CI) 1.02-1.17, p = 0.013) and epicardial adiponectin levels (OR 0.98, 95% CI 0.97-1.00, p = 0.054) as independent predictors of postoperative AF. Conclusions: Increased epicardial adiponectin is associated with maintenance of sinus rhythm following cardiac surgery. This reinforces the inflammatory hypothesis in the pathogenesis of postoperative AF and may represent a novel therapeutic target for its effective prevention.
Obesity, defined by an excess of adipose tissue, is often associated with the development of various metabolic diseases. The increased and inappropriate deposition of this tissue contributes to ...hyperglycemia, hyperlipidemia, insulin resistance, endothelial dysfunction and chronic inflammation. Recent evidence suggests that factors expressed and secreted by the adipose tissue, adipokines, may contribute to the development of these abnormalities by mechanisms including inhibition of adipogenesis, adipocyte hypertrophy and death, immune cell infiltration and disruption of tissue metabolism. The presence of adipokine receptors in adipocytes renders these cells available to autocrine and paracrine effects of adipokines. In this review the reported local effects of adipokines on adipose tissue structure, inflammation and regulation of metabolic functions, in the face of over-nutrition and consequent obesity, are outlined. Elucidating the local regulation of white adipocyte development and function could help in the design of effective, tissue-specific therapies for obesity-associated diseases.
The n‐3 polyunsaturated fatty acids, especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), exert hypolipidemic effects and prevent development of obesity and insulin resistance in ...animals fed high‐fat diets. We sought to determine the efficacy of α‐substituted DHA derivatives as lipid‐lowering, antiobesity, and antidiabetic agents. C57BL/6 mice were given a corn oil‐based high‐fat (35% weight/weight) diet (cHF), or cHF with 1.5% of lipids replaced with α‐methyl DHA ethyl ester (Substance 1), α‐ethyl DHA ethyl ester (Substance 2), α,α‐di‐methyl DHA ethyl ester (Substance 3), or α‐thioethyl DHA ethyl ester (Substance 4) for 4 months. Plasma markers of glucose and lipid metabolism, glucose tolerance, morphology, tissue lipid content, and gene regulation were characterized. The cHF induced obesity, hyperlipidemia, impairment of glucose homeostasis, and adipose tissue inflammation. Except for Substance 3, all other substances prevented weight gain and Substance 2 exerted the strongest effect (63% of cHF‐controls). Glucose intolerance was significantly prevented (∼67% of cHF) by both Substance 1 and Substance 2. Moreover, Substance 2 lowered fasting glycemia, plasma insulin, triacylglycerols, and nonesterified fatty acids (73, 9, 47, and 81% of cHF‐controls, respectively). Substance 2 reduced accumulation of lipids in liver and skeletal muscle, as well as adipose tissue inflammation associated with obesity. Substance 2 also induced weight loss in dietary obese mice. In contrast to DHA administered either alone or as a component of the EPA/DHA concentrate (replacing 15% of dietary lipids), Substance 2 also reversed established glucose intolerance in obese mice. Thus, Substance 2 represents a novel compound with a promising potential in the treatment of obesity and associated metabolic disturbances.
Consumption of a high-carbohydrate diet has a critical role in the induction of weight gain and obesity-related pathologies. This study tested the hypothesis that a carbohydrate-rich diet induces ...weight gain, ectopic fat deposition, associated metabolic risks and development of non-alcoholic fatty liver disease (NAFLD), which are partially reversible following carbohydrate reduction. Sprague Dawley (SD) rats were fed a carbohydrate-enriched cafeteria diet (CAF) or normal chow (NC) ad libitum for 16-18 weeks. In the reversible group (REV), the CAF was replaced with NC for a further 3 weeks (18-21 weeks). Animals fed the CAF diet showed significantly increased body weight compared to those fed NC, accompanied by abnormal changes in their systemic insulin and triglycerides, elevation of hepatic triglyceride and hepatic steatosis. In the REV group, when the CAF diet was stopped, a modest, non-significant weight loss was associated with improvement in systemic insulin and appearance of the liver, with lower gross fatty deposits and hepatic triglyceride. In conclusion, a carbohydrate-enriched diet led to many features of metabolic syndrome, including hyperinsulinemia, while a dietary reduction in this macronutrient, even for a short period, was able to restore normoinsulinemia, and reversed some of the obesity-related hepatic abnormalities, without significant weight loss.
Coronavirus disease of 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has sparked a global pandemic with severe complications and high morbidity rate. ...Neurological symptoms in COVID-19 patients, and neurological sequelae post COVID-19 recovery have been extensively reported. Yet, neurological molecular signature and signaling pathways that are affected in the central nervous system (CNS) of COVID-19 severe patients remain still unknown and need to be identified. Plasma samples from 49 severe COVID-19 patients, 50 mild COVID-19 patients, and 40 healthy controls were subjected to Olink proteomics analysis of 184 CNS-enriched proteins. By using a multi-approach bioinformatics analysis, we identified a 34-neurological protein signature for COVID-19 severity and unveiled dysregulated neurological pathways in severe cases. Here, we identified a new neurological protein signature for severe COVID-19 that was validated in different independent cohorts using blood and postmortem brain samples and shown to correlate with neurological diseases and pharmacological drugs. This protein signature could potentially aid the development of prognostic and diagnostic tools for neurological complications in post-COVID-19 convalescent patients with long term neurological sequelae.
•Neurological protein signature of severe COVID-19.•Validation of Qatar severe COVID-19 neurological protein signature in independent cohorts at protein and RNA levels.•Neurological proteins signature of severe COVID-19 corelates with Neurological diseases such as Nerve Injury, Parkinson's disease, Alzheimer's disease, Amyotrophic lateral sclerosis, and Schizophrenia.•Neurological proteins signature of severe COVID-19 corelates with pharmacological drugs such as risperidone.•The risk of developing chronic neurological diseases in post-severe COVID-19 patients.
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