ACTA ENDOCRINOLOGICA (BUC)

Objective. The aim of this study was to evaluate the impact of sperm DNA fragmentation (SDF) on fertilization rate, and sperm nuclear decondensation after intracytoplasmic injection of sperm (ICSI) into cumulus-free germinal vesicle (GV) oocytes from stimulated cycles. Methods and study design. After the retrieval of oocytes, the GV oocytes were cultured for 24 h. Oocytes that liberated polar bodies were injected with processed semen, and were used to evaluate SDF level. Based on SDF, the data were categorized into two groups. Group I in which SDF≤ 30% and group II in which SDF > 30%. Fertilization (presence of two pronuclei) was checked 16-19 h after ICSI. Unfertilized oocytes were stained by Hoechst 33258 and examined to evaluate the undecondensed sperm head in oocyte. The rates of maturation, fertilization in fertilized IVM oocytes and the percentage of undecondensed sperm in the unfertilized oocytes were assessed according to SDF. Results. Out of 146 GV oocytes that were subjected to IVM, 101 (69 %) developed to metaphase II. The fertilization rate of IVM oocytes in group II was significantly lower than that in group I (P < 0.05). Moreover, group I, had 25 % of their unfertilized oocytes containing condensed sperm, while group II had a significantly higher number (53 %) of unfertilized oocytes containing condensed sperm (P < 0.05). Conclusion. SDF had a negative effect on the rate of fertilization in matured in vitro GV oocytes and could lead to an increase in the percentage of undecondensed sperm in IVM oocytes from stimulated cycles.

Objective. The aims of this study were to compare the effect of telmisartan and valsartan on the blood pressure and insulin sensitivity and adiponectin levels in hypertensive patients with metabolic syndrome.\r\nPatients and Methods. One hundred twenty male patients who met the criteria for metabolic syndrome defined as in ATP III enrolled in the sudy. All patients were randomized to receive treatment with telmisartan 80 mg (n=70) or valsartan 160 mg (n=50) once daily for 6 months. Serum insulin concentration was measured by chemiluminescence, plasma levels of adiponectin and visfatin by Enzyme linked immunosorbent assay kit (ELISA). Insulin resistance (IR) was estimated using the homeostasis model assessment (HOMA).\r\nResults. Mean HOMA-IR and adiponectin and visfatin levels were measured 3.00?0.9 and 5.93?0.3 mgr/ml and 23.45?6.1 mgr/ml in patients before starting treatment of telmisartan, respectively. They were changed to 2.4?0.5 and 6.71?0.5 mg/ml and 21.40?5.0 mg/ml at a 6 months period. In valsartan group, mean HOMA-IR and adiponectin and visfatin levels were measured 2.9?0.5 and 5.50?0.9 mg/ml 19.05?3.9 mg/ml before treatment, respectively. After a 6 months period, mean HOMA-IR and adiponectin and visfatin levels were found 3.00?0.9 and 6.24?0.7 mg/ml and 20.76?4.5 mg/ml, respectively. Conclusion: Telmisartan produced significant reductions from baseline in HOMA-IR and insulin levels, whereas valsartan did not. Both telmisartan and valsartan did no changed serum visfatin levels but they increased serum adiponectin concentrations by RAS blockade.

12 adult male and 12 female Wistar albino rats were used to measure the levels of the serum follicle stimulating hormone (FSH), the luteinizing hormone (LH), the estradiol (E2) and the progesterone in female rats, as well as the FSH, the LH and the testosterone in male rats, following 8-hour daily inhalation exposure to gasoline vapours for 6 weeks, to assess the effect of the vapours on the reproductive integrity of experimental animals. The results showed that the levels of serum FSH and LH obtained for female rats in the test group (3.48?0.03 and 19.97?1.20mIU/ml, respectively) were insignificantly lower (p>0.05), compared respectively to the levels obtained for female rats in the control group (3.62?0.12 and 20.06?0.23mlU/ml, respectively), whereas the levels of serum estradiol and progesterone obtained for female rats in the test group (25.02?2.32pg/ml and 0.18?0.02mg/ml, respectively) were significantly lower (p<0.05), compared respectively to the levels obtained for female rats in the control group (39.84?3.64pg/ml and\r\n0.33?0.02mg/ml, respectively). On the other hand, the levels of serum FSH and LH obtained for male rats in the test group (2.87?0.21 and 3.09?0.32mIU/ml, respectively) were insignificantly higher (p>0.05), compared to the levels obtained for male rats in the control group (2.75?0.14 and 2.93?0.12mIU/ml, respectively).At the same time, the level of serum testosterone in the male test rats (7.82?2.17ng/dl) was significantly higher (p<0.05), compared to the level in the male control rats 4.66?1.85ng/gl. The results of this study showed that the adverse effect of inhalation exposure to gasoline fumes on the reproductive integrity in rats is sex-dependent, with the females being more vulnerable.

Background. Methotrexate (MTX) is used in the treatment of neoplastic disorders. MTX action in non-neoplastic diseases still remains obscure. Female reproductive cells are fast proliferating like cancer cells. Hence, it is important to\r\nidentify markers affected by MTX in the reproductive tract.\r\nAim. Investigate MTX effect on energy metabolism marker glycogen and the protective role of leucovorin (LCN) and\r\nwithdrawal of MTX treatment in the ovary, oviduct, uterus, cervix and vagina of rats.\r\nAnimals and Methods. Rats with regular oestrous cycle were randomly divided into five groups (n=6) as follows: Control, MTX LD (low dose), MTX HD (high dose), MTXHD+LCN (leucovorin), and MTXHD+WD (withdrawal): 20 days withdrawal. Animals were treated once per day intramuscularly for 20 days. Rats were sacrificed on day 21. MTXHD treatment was withdrawn for additional 20 days and\r\nanimals sacrificed on day 41. Ovary, oviduct, uterus, cervix and vagina were used for glycogen analysis.\r\nResults. The present study explored the effect of MTX and LCN on glycogen content in the ovary, oviduct, uterus, cervix and vagina of rats. MTX significantly (P<0.001) inhibited glycogen content in ovary, oviduct, uterus, cervix and vagina of rats, which was dose dependent. LCN\r\nsupplementation and withdrawal of MTX treatment, partially recovered such an effect.

Aim. We investigated the relationship between irisin concentrations and glycemic control, body composition and anthropometric measures in children with type 1 diabetes mellitus. Methods. The study involved 40 subjects with T1DM prospectively. Glycemic control was evaluated. Body composition was analyzed with a bioimpedance analyzer (BIA). Serum irisin concentrations were measured using an ELISA kit. Results. Irisin levels were found higher in BMI <17 kg/m2 group (p=0.002) compared to BMI >17 kg/m2. Irisin level was negatively correlated with weight, height, BMI, fat free mass, skeletelal muscle mass, basal metabolic rate (r= -0.40, p= 0.011; r=-0.32, p=0.046; r=-0.366, p= 0.022; r=-0.423, p= 0.007; r=-0.430, p=0.006; r=-0.416, p=0.009, respectively); there was a strong correlation between LDL-C and irisin levels (r=0.367, p=0.02). In multivariate linear regression analyses model, irisin concentrations were correlated with weight (ß-coefficient= - 0.391, p= 0.015). LDL-C is associated, but not correlated significantly with irisin levels, (ß-coefficient =0.272, p=0.084). Conclusion. As a result, weight and LDL-C were the predictors of circulating irisin. To our knowledge, this study is the first examining association between irisin levels and body composition comprehensively, in children with type 1 diabetes mellitus.

Introduction. Purslane (Portulaca oleracea L.) has antioxidant effects. The aim of this study was to evaluate the pancreas protective effect of Purslane hydroalcoholic extract in D-galactose induced aging female mouse model. Methods. In this experimental study, 72 adult female mice (30 – 35 g) were obtained and divided into 6 groups: control, Purslane hydroalcoholic extract, D-galactose, D-galactose + Purslane hydroalcoholic extract, Aging, Aging + Purslane hydroalcoholic extract. The aging model induced by subcutaneous injection of D-galactose for 45 consecutive days, and Purslane hydroalcoholic extract was orally gavaged in the last 21 days. 24 hours after the last drug and extract administrations, serum samples and pancreas tissues were removed for biochemical and histological assessments. Results. Glucose decreased in the Purslane, D-galactose + Purslane and Aging + Purslane groups (p<0.05). Insulin and HOMA-IR increased in D-galactose and, Aging groups (p<0.05). Furthermore, administration of hydroalcoholic extract of Purslane improved these parameters in D-galactose and Aging treated mice (p<0.05). Diameter of pancreatic islets decreased in Aging and D-galactose groups and Purslane hydroalcoholic extract administration improved this variable. Conclusions. The present results show that Purslane has pancreas protective effects via its hypoglycemic and insulin resistance reducing activity.

Objective. In this study we investigated the effect of dorsomedial hypothalamus (DMH) neuropeptide Y (NPY) knock-down on hepatic insulin sensitivity in high-fat (HF) diet-fed rats. Methods. Forty-eight Sprague-Dawley rats were randomly assigned to receive bilateral DMH injections of adeno-associated virus AAVshNPY or AAVshCTL and then accessed to regular chow. Five weeks after viral injection, half rats in each group were given access to the HF diet. At 16 weeks, rat livers were collected. Insulin receptor substrate-1 (IRS-1) and phosphoinositide 3-kinase (PI3K) mRNA expression was measured by qRT-PCR. Blood glucose levels were measured by the oxidase method, serum insulin, triglyceride, and TC levels were measured by Elisa. Pathological changes in the liver were assessed by hematoxylin-eosin (HE) staining. AKT, p-AKT, and GSK-3 levels were measured by western blotting. Results. Compared with AAVshCTL-injected rats, AAVshNPY-injected rats showed a significant decrease in blood glucose concentrations; serum insulin, triglyceride, and TC; HOMA-IR; and IRS-1 and PI3K mRNA levels (P<0.05). ISI, GSK-3, and p-AKT levels were significantly increased (P<0.05). HE staining showed that AAVshNPYinjected rats fed the HF diet had mild fatty degeneration. Conclusion. These results suggest that DMH NPY knock-down improves hepatic insulin sensitivity in HF diet-fed rats by activating the hepatic PI3K/AKT insulin signalling pathway.

Context. Abnormally increased hepatic glucose production contributes to hyperglycemia in diabetes. Interventions that suppress hepatic gluconeogenesis should be beneficial in improving glycemic control in patients with diabetes. Objectives. It has been suggested that hepatic FTO is involved in glycemic control by regulating gluconeogenesis. Both FTO and activating transcription factor 4 (ATF4) positively regulate the expression of gluconeogenic genes in the liver, suggesting the possibility that ATF4 mediates the stimulatory effect of FTO on hepatic gluconeogenesis. The present study aimed to determine the effect of altered expression or activity of FTO on Atf4 and gluconeogenic gene expression in hepatocyte cells. Methods. Mouse hepatocyte AML12 cells were treated with the FTO inhibitor rhein or transfected with an FTO-expressing plasmid. Levels of gluconeogenic glucose- 6-phosphatase (G6pc) and Atf4 mRNA and protein were measured. Results. Rhein treatment significantly reduced G6pc mRNA levels as well as Atf4 mRNA and protein levels. Conversely, enhanced FTO expression caused an increase in G6pc and Atf4 mRNA levels. Conclusions. These findings support the hypothesis that hepatic FTO participates in the regulation of hepatic gluconeogenic gene and ATF4 expression. Reducing the activity of the hepatic FTO-ATF4 pathway may be beneficial in reducing hepatic glucose production and ameliorating hyperglycemia in diabetes.

Context. With more studies investigating effects of high serum lipid levels, new findings are emerging regarding the damage these biomolecules may cause. Aim. In this study we aimed to find a relation between neuropathy and hypertriglyceridemia in patients with metabolic syndrome (MS). Material and methods. One hundred and twenty subjects (Ninety subjects with metabolic syndrome and 30 healthy controls) were included in the study. Subjects with MS were divided into three groups. HbA1C levels of the subjects were < 5.7% in group A, ≥ 5.7% - < 6.5% in group B, and ≥ 6.5% - < 8.0% in group C. Pin-Prick test and Semmes- Weinstein Monofilament were used for neurological examination. Electromyography was performed to patients with neuropathy to support the diagnosis. Results. Neuropathy prevalence was found to be higher in the subjects with metabolic syndrome compared to control group. (9.9 %; 16.65 %; 23.31 % vs. 3.3%; in group A, group B, group C vs. healthy control group respectively) (p=0.003 for group A, p=0.0002 for group B, p=0.0002 for group C). There was an association between triglyceride levels and neuropathy in group C. Conclusion. Patients with MS may have more neuropathy risk than we estimate.

Background. Chronic kidney disease (CKD) and type 2 diabetes mellitus (T2DM) are associated with disturbed mineral homeostasis and serum bone biomarkers. The interplay between T2DM and CKD on serum bone turnover markers (BTM) is unclear. Our aim was to describe the BTM in patients with T2DM, CKD or both. Methods. In this observational, single-centre, prospective study, we included 320 patients over 40 years, divided into four groups: T2DM and normal kidney function (n=142), T2DM and CKD (n=36), CKD and normal glucose metabolism (n=29) and healthy controls (n=113). We excluded patients treated for osteoporosis and with secondary osteoporosis. Patients were compared by age, levels of glycated hemoglobin, PTH, alkaline phosphatase, osteocalcin (OC), CTx and 25 OH vitamin D. Results. Univariate analysis showed that GFR correlated significantly with PTH (r=0.37), OC (r=0.43) and CTX (r=0.45) in the diabetes group but only with PTH (r=0.34) in the non-T2DM group. Multivariate analysis showed that GFR remained significantly correlated with the same bone markers even after adjustment for age, sex or 25(OH)D levels. Diabetics seem to have lower levels of alkaline phosphatase (68±22.1 U/L) and CTX (0.37±0.24 ng/mL) than those without diabetes (76.7±29.6. U/L and 0.5±0.19 ng/mL, respectively). There was no correlation between BTM and glycated hemoglobin. Conclusions. Bone turnover markers correlate with GFR, particularly in patients with T2DM. However, alkaline phosphatase is lower in T2DM than in non-T2DM.