ACTA ENDOCRINOLOGICA (BUC)

Introduction. The recall rate in congenital hypothyroidism (CH) newborn screening programs depends on several factors such as primary screening strategy, sample collection guidelines, and cut-off. A recall rate of 0.05% is recommended but recall rates of 0.01-13% are reported worldwide. Objective. The aim of the study was to assess the recall rate in association with the age of the newborn at sample collection and with a lower cut-off. Design. This was a retrospective correlational study concerning TSH results in CH screening program. Subjects and methods. All newborns from a tertiary center in Targu Mures, Romania between 2013-2018 were included. Four groups were created and a correlation test between TSH median value and age in days was performed. The recall rate was calculated using three cut-off levels for 8182 TSH results performed in 2018. Results. 90% of the DBS were collected in days 3-5 after birth and 1/79 live births from this group had TSH above the lower cut-off used (10 mUI/L). 2% of the samples were collected in less than 48 hours after birth and 1/21 live birth from this group had TSH > 10 mUI/L. The recall rate in our center in 2018 was 0.08%, higher than the recommended rate of 0.05%. With the cut-off value recommended by the Health Ministry a 0.03% recall rate was obtained. Conclusions. Sample collection in less than 48 hours increases the recall rate in CH screening with TSH primary testing strategy. A lower cut-off raises the recall rate with a higher rate of false-positive cases (94.12%) but with 100% negative predictive value.

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.

Context. Recent evidence has stressed that many proinflammatory factors are particularly conducive to the progression of diabetic nephropathy, but the mechanisms underlying the changes are poorly understood. Objective. The purpose of this study was to investigate if up-regulated expression of both phospholipase A2 (PLA2) and cyclooxygenase-2 (COX-2) is involved in renal damage and micro-inflammatory state in streptozotocin-induced diabetic rats. Animals and methods. Sixteen Sprague Dawley rats were randomly divided into 2 groups: control group and diabetes group. Animals in diabetes group were treated with intraperitoneal injection of streptozotocin. Eight weeks later, rat renal tissue was studied with light and transmission electron microscopes, and PLA2 and COX-2 and their mRNA expression were examined by immunohistochemistry and reverse transcription polymerase chain reaction, respectively. Results. The renal pathological lesions in diabetes group were obvious, including increased amounts of mesangial matrix, thickening of the glomerular and tubular basement membranes and fusion and effacement of the adjacent podocyte foot processes. Infiltrating inflammatory cells were observed in the tubules. Compared with control group, the expression of cytosolic PLA2 and COX-2 was significantly increased in diabetes group. Conclusions. It uncovers that the PLA2-COX-2 pathway may lead to renal inflammation associated with renal damage in streptozotocin- induced diabetic rats.

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.

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.

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.

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.

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.

Objective. We aimed to investigate whether lifestyle and body fat mass have an impact on the occurence of nonfunctional adrenal incidentalomas (NFAI). Methods. 100 patients with NFAI were included . 50 people constituted the control group. Physical activities of these groups were evaluated (using the International Physical Activity Questionnaire), smoking status was determined, anthropometric measurements were made. Body fat mass, fat percentage, total body water and fat free mass were measured using bioelectrical impedance method. Results. Body mass index (BMI), waist, hip, neck circumference, total body fat percentage and fat mass and smoking rate were found to be statistically higher in the patient group. Physical activities did not differ significantly. When a subgroup with similar age and BMI among was created, waist circumference and total fat mass were again significantly higher in the patient group. There was a significant positive correlation between the size of the adrenal mass and waist, neck circumference, BMI, and cortisol after 1 mg dexamethasone suppression test. Conclusion. The increase in the fat mass may have an impact on the development of NFAI. Although the patients were regarded as nonfunctional, suppressibility of the cortisol decreases as the mass size of the incidentaloma increases.

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.