ACTA ENDOCRINOLOGICA (BUC)

Introduction. Menopause increases the risk of cardiovascular disease in women. The aims of the present study were to evaluate the effects of swimming training on cardiac histology and expression of miR-29 and IGF-1 in the ovariectomized rats. Materials and methods. Thirty female Wistar rats were divided into sham and ovariectomized groups: sedentary control (OVX) and trained with 8 weeks exercise (OVX.E). On 57th day, blood was collected and used for lipid profile measurement. In addition, heart tissue was analyzed by reverse transcription–polymerase chain reaction for IGF- 1 mRNA and miR-29, and studied for histopathological changes. Results. Ovariectomy significantly decreased miR- 29 and IGF-1 expression in the heart compared to sham animals group (p<0.05). Exercise training increased miR-29 and IGF- 1 expression in the trained rats and improved histology and lipid profile compared with OVX group (p<0.05). Conclusion. Estrogen deficiency could lead to cardiac fibrosis through deregulation miR-29 and IGF-1 expression. The findings of the current study suggests a protective effect of exercise on heart against fibrotic changes in ovariectomized rats and support a potential preventive value of exercise in improving cardiac function after menopause.

Phenylketonuria (PKU) is the most frequent inherited amino acid metabolic disorder, and it may also be treated by dietary means. The determination of phenylalanine (Phe) levels in the blood plasma is important not only in early diagnostic, but also in monitoring the treatment of PKU. Purpose. The aim of our work was to develop a simple, sensitive and highly accurate procedure to determine the plasma concentration of Phe. Procedure. The measurement of plasma Phe concentration involves two steps: a) separation of plasma (from the blood taken on heparin), isolation and preparation of a concentrated solution of amino acids (by ion-exchange column chromatography on Dowex-50X8 and evaporation of the eluate in vacuum at 40˚C), and b) determination of Phe concentration in the solution of amino acids by HPLC. This analysis was performed using a Dionex Ultimate 3000 instrument equipped with a Ultimate 3000 diode array detector (DAD). The values of Phe concentration in the plasma of several patients were calculated using a calibration curve made with standards of Phe (dilutions of a stock solution of 50 mg/ dL). The measurements in duplicate (plasma Phe) or a greater number of samples from the same concentration of standards of Phe showed extremely small sample to sample differences. Concentrations as low as 0.2 mg/dL could be determined. Conclusion. The whole procedure presented here is relatively simple, rather inexpensive, however very sensitive and highly accurate. Consequently, it is very adequate for confirming the diagnosis of PKU in patients with neonatal hyperphenylalaninemia, as well as for monitoring the plasma concentration of Phe in patients with PKU.

In response to increasing interest of the European Commission on large-scale\r\ngenotyping for complex diseases, including variability in ethnic minorities in\r\nEurope (HEALTH-2009-4.3.3-1), at the end of 2008 we composed the\r\nHAPLOGENDIS consortium with partners from Russia and European countries. A\r\nfirst program (SICA) was proposed in cooperation with Russian Federal Agency for\r\nScience and Innovation, focusing on comparative population genetics on diseases\r\naccompanied by insulin resistance. Beside the specificity in analyzing the human\r\ngenome with SNP (single nucleotide polymorphism) and defining haplotype\r\nstructure of genes, the program rises new hypotheses which directly link\r\ncolonization of Europe at the Neolithic period from Eastern Ukraine or Anatolia\r\nwith the development of agriculture and major dietary and life style changes that\r\nmay have an impact on the genome. Although there will be many occasions to\r\nreview both genetic and clinical detailed aspects, this short note will expose some\r\nunifying ideas that joint these partners.

Context. MicroRNAs (miRNAs) are short noncoding RNAs involved in posttranscriptional regulation of gene expression that influence various cellular functions including glucose and lipid metabolism and adipocyte differentiation. Objective. The aim of this study was to evaluate the levels of miR-34a and miR-149 and their relationship with metabolic parameters in obese children and adolescents. Design. Seventy children and adolescents were enrolled in the study. Plasma levels of microRNAs were evaluated by real-time PCR using SYBR green and analyzed by ΔCt method. Plasma concentrations of visfatin and insulin were measured by ELISA method. Glucose and lipid profile were determined colorimetrically. HOMA-IR was calculated and used as an index of insulin resistance (IR). Results. miR-34a was significantly lower in subjects with insulin resistance compared to obese children with normal insulin sensitivity. There was an inverse relationship between miR-34a levels and both insulin and HOMA-IR. On the other hand, miR-149 was significantly correlated with visfatin. There was no significant difference in miR-34a and miR-149 between obese and normal weight subjects. Conclusions. miR-34a is associated with insulin and HOMA-IR and thus seems to be involved in IR. miR- 149 is inversely associated with visfatin levels which could be indicative of anti-inflammatory effect of this miRNA.

Objective. In our study, we aimed to investigate the levels of irisin, nesfatin-1 and the relationship between levels of these relatively new molecules with cardiometabolic risk markers; carotid intima-media thickness (CIMT), epicardial adipose tissue (EAT) thickness in patients with nonfunctional adrenal incidentaloma (NFAI). Materials and Methods. Patients with NFAI (n=59) and age, sex and body mass index-matched healthy control subjects (n=59) were enrolled in this study. Serum glucose, insulin, C-reactive protein (CRP), lipid, irisin and nesfatin-1 levels and echocardiographic CIMT and EAT thickness measurements were performed in patients and controls. Results. The irisin level was 17.58 ± 4.38 pg/mL in the NFAI group, significantly higher (p<0.001) than 14.03 ± 4.03 pg/mL in the control group. Nesfatin-1 level was significantly lower in the NFAI group 194.98 ± 119.15 pg/ mL ((p < 0.001)) versus 303.48 ± 200.78 pg/mL in the control group. A positive correlation was found between irisin and nesfatin-1 levels and CIMT and EAT thickness in the NFAI group. Conclusions. In our study, we found that irisin level was higher and nesfatin-1 level was lower in patients with NFAI, and both irisin and nesfatin-1 levels were associated with CIMT and EAT thickness in NFAI patients.

Gastrointestinal complaints are common among diabetic patients. The gastrointestinal tract contains melatonin. The binding sites of melatonin have been identified in all GIT tissues. Melatonin can modify activities of the gut and liver. The aim of this study was to evaluate the possible protective effects of melatonin against gastric motility and secretary responses in Streptozotocin-induced diabetes in rats. Methods. Streptozotocin was injected intraperitoneally at a single dose of 60 mg/kg for diabetes induction. One week after inducing diabetes, Melatonin (5, 10, 20 mg/ kg/day, IP.) was injected for 14 days. Gastric acid and mucus were measured in all animals by chemical methods. Gastric motility was investigated by powerlab system. Results. Streptozotocin induced a significant increase in blood glucose levels (p<0.001) and significant decrease in gastric acid, mucus, motility and body weight in diabetic groups. Treatment of diabetic rats with melatonin significantly reduced blood glucose (p<0.001) and increased gastric mucus (p<0.001) and motility (p<0.01 and p<0.05 in groups 4 and 5 respectively) with no effect on body weight and gastric acid concentration. Conclusion. These data suggested that melatonin treatment has a therapeutic effect on diabetic gastrointestinal disturbances by reduction of serum glucose and increasing gastric motility and gastric mucus levels, but no effect on gastric acid and body weight.

Objective. The aim of this study was to investigate the relationship of Claudin-5, Apelin, Tumor Necrosis Factor Alpha (TNF-α) expression, and body mass index (BMI) of cholecystectomies. Materials and methods. Sixty-eight paraffin embedded cholecystectomy specimens diagnosed as chronic cholecystitis were collected in the Pathology Department of the Training and Research Hospital between 2015-2017. The samples were stained with Apelin, Claudin-5 and TNF-α. The immunohistochemical study was carried out using the system in an automatic staining machine. Results. There was a significant positive correlation between BMI and TNF-α staining (p=0.010). This result indicated that the degree of staining increased together with BMI. When age, BMI, and the other biochemical parameters were evaluated, a significant correlation was found between BMI and blood glucose only (p=0.029); correlations of BMI with the other parameters were not statistically significant. Conclusion. Although there is no relationship between inflammation and BMI with Claudin-5 and Apelin in this study, there is a significant relationship between BMI and TNF-α.

Background. Pomegranate is a rich source of many polyphenolic compounds including ellagitannins (punicalagin, punicalin and others). Aim. The effects of punicalagin and punicalin on adipogenesis were investigated in this study. Materials and Methods. To examine the effect of punicalagin and punicalin on adipocyte differentiation, various concentrations of punicalagin and punicalin (2- 10 μM) were applied to differentiated 3T3-L1 cells. Glyceraldehyde-3-phosphate dehydrogenase (GPDH) activity, Oil red O staining, intracellular triglyceride levels, and gene expressions of transcription factors (Peroxisome proliferator-activated receptor-γ (PPARγ), CCAATenhancer- binding proteins-α (C/EBPα), Sterol regulatory element-binding protein 1c (SREBP-1c)) and lipolysisassociated genes (hormone-sensitive lipase (HSL), Perilipin A, tumor necrosis factor-α (TNF-α)) were examined in order to investigate the effects of punicalagin and punicalin on adipocyte differentiation. Results. Punicalagin and punicalin applications caused a continuous decrease in cell size and intracellular triglyceride accumulation. GPDH activity and transcription gene expressions decreased significantly in groups that were applicated punicalagin and punicalin at high concentrations. Punicalagin, but not punicalin, down-regulated the expression of HSL and perilipin A and up-regulated the expression of TNF-α in a dose-dependent manner. In conclusion, both punicalagin and punicalin were able to inhibit the adipocyte differentiation.

Background. Hashimoto’s thyroiditis is in coexistence with many autoimmune disorders, especially celiac disease. There are a limited number of studies evaluating the prevalence of celiac-related antibodies in patients with Hashimoto’s thyroiditis. Objective. This study aimed to further investigate the prevalence of undiagnosed celiac disease in patients with Hashimoto’s thyroiditis and the relationship between these two autoimmune disorders in these patients Subjects and methods. This study was performed on 82 women aged 20-50 years including 40 patients with Hashimoto’s thyroiditis and 42 healthy age-matched individuals. Anthropometric assessments were performed and biochemical parameters including thyroid hormones (TSH, T3 and T4), antithyroid antibodies, anti-tissue transglutaminase and anti-gliadin antibodies were measured by enzyme linked immunosorbent assay (ELISA). Results. The prevalence of IgG and IgA anti-tissue transglutaminase antibodies and IgA anti-gliadin antibody was higher in Hashimoto’s thyroiditis patients compared with control group (15% vs. 7%, 22.5% vs. 17% and 15% vs. 12% respectively). In ordinal regression model, serum IgG anti-tissue transglutaminase and IgA anti-gliadin antibodies were significant predictors of antithyroid antibodies in patients with Hashimoto’s thyroiditis (P < 0.05). A significant relationship between serum TSH and IgG antigliadin antibody were also found (P = 0.003). Conclusion. To our findings, a high prevalence of anti-tissue transglutaminase and IgA anti-gliadin antibodies and their positive relationship with antithyroid antibodies in patients with Hashimoto’s thyroiditis were reported. These findings further warrant the need for interventions to reduce the prevalence of these antibodies in Hashimoto’s thyroiditis for preventing the occurrence of celiac disease in these patients.

Context. Endothelial dysfunction and diabetic cardiomyopathy are critical complications of diabetes. Gallic acid (GA) plays a significant role in cardiovascular disorders resulted from diabetes. In addition, increased plasma miR-24, miR-126 associated with endothelial dysfunction. Aim. The current study was designed to assess the effects of GA on plasma miR-24, miR-126 levels in the diabetic rats. Animals and Methods. Adult male Sprague-Dawley rats were divided into three groups (n=8): control (C), diabetic (D) and diabetic group treated with GA (D+G, 25 mg/kg, by gavage) for eight weeks. The blood glucose level, body weight, lipid profile, blood pressure, plasma miR-24 and miR-126 levels, antioxidant and inflammatory biomarkers were measured. Results. The plasma levels of miR-24, miR-126, body weight, high-density lipoprotein cholesterol (HDL-c), total anti-oxidant capacity (TAC) and the systolic blood pressure significantly reduced and blood glucose, total cholesterol (TC), triglycerides (TG), very low-density lipoprotein cholesterol (VLDL-c), malondialdehyde (MDA), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α) and low-density lipoprotein cholesterol (LDL-c) significantly elevated among the diabetic rats compared with the control group. However, GA restored body weight, blood pressure, TC, TG, VLDL-c, TNF-α, miR- 126, blood glucose, HDL-c, MDA, TAC, miR-24 and IL-6 among the GA treated rats compared with the diabetic group. Conclusion. GA improves inflammation, oxidative stress and hypotension result from diabetes. These protective effects are probably mediated via increasing plasma miR-24 and miR-126 levels.