ACTA ENDOCRINOLOGICA (BUC)

Fine needle aspiration biopsy (FNAB) was designed as a rapid and cost efficient tool in the morphological diagnosis of thyroid nodules. Provided by efficient sampling especially by the wide scale use of ultrasound guidance, by the on-site evaluation of the quality of sampling, the FNAB, as a diagnostic tool, still have an undetermined ratio of diagnosis ranging from 5 to 40 % according to the experience of the examinator. About what are the critical points defining the performance of FNAB in thyroid nodules, and where is the place of molecular biology. I will insert some short comments: the critical steps in performing FNAB are the sampling, the interpreting and the good communication of the diagnosis to the clinician.

Background. Hyperuricemia is associated with non-alcoholic fatty liver disease (NAFLD). Aim. We therefore aimed at evaluating the influence of allopurinol on the course of NAFLD in rats. Study Design. We divided 21 mature albino Sprague Dawley rats into three groups: controls (n = 7, normal diet for 12 weeks); NAFLD rat models (by feeding water containing 30% fructose for first 8 weeks) treated with allopurinol subsequently for the next 4 weeks (n = 7); and similar case treated with placebo (saline) subsequently for the next 4 weeks (n = 7). Methods. We compared the histopathological scores, IL-1 and IL-2 immunoexpression levels across the groups. Liver histopathological score was determined by observing the steatosis (the percentage of liver cells containing fat): <25% = 1+, 25% - 50% = 2+, 51% - 75% = 3+, >75% = 4+; inflammation and necrosis: 1 focus per low-power field = 1+; and 2 or more foci = 2+. The number of liver IL-1 and IL-2 positive cells was measured by systematically scoring at least 100 hepatocyte cells per field in 10 fields of tissue sections by a magnification of 100. Results. Xanthine oxidase (XO) activity and lipid peroxidation was significantly different in the allopurinol group compared to the saline group (XO; 0.098 ± 0.006 mU/mg vs. 0.162 ± 0.008 mU/mg, p = 0.01, 0.116 ± 0.040 nmol malondialdehyde/mg versus 0.246 ± 0.040 nmol malondialdehyde /mg, p = 0.01). The allopurinol group had lower histopathological scores, IL-1 and IL-2 immunoexpression levels in the liver compared to the saline group (2.13 ± 0.35 against 5.45 ± 0.24, p = 0.003, IL-1; 5.76 ± 0.43 against 12.85 ± 3.26, p = 0.023, IL-2; 8.55 ± 1.14 against 56.23 ± 7.12, p = 0.002). Conclusions. Allopurinol has a therapeutic role against the progression of NAFLD of the rats.

-

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.

Thyroid hormones play a crucial role in the regulation of the mitochondrial oxidative\r\nmetabolism. Hyperthyroidism caused by the acceleration of the energy metabolism leads to\r\nthe occurrence of cellular oxidative stress.\r\nThe aim is to evaluate the pro-oxidant / antioxidant balance and the effect of vitamin\r\nE supplementation in damage caused by the excessive administration of thyroid hormones.\r\nMaterials and Methods. White, male Wistar rats were used in the study. Thirty male\r\nWistar rats were divided into three groups (1:control group, 2:animals treated with LThyroxine\r\n10 &#956;g/animal/day for 30 days, 3:L-Thyroxin treated rats protected with vitamin\r\nE 10 mg/animal/day). Malondialdehyde (MDA), the marker of lipid peroxidation, carbonyl\r\nproteins, SH groups, glutathione (GSH) and superoxide dismutase (SOD) were determined\r\nfrom the serum, while MDA, carbonyl proteins, SH groups and GSH were determined from\r\nthe thyroid tissue homogenates.\r\nThe results showed increased levels of carbonyl proteins (1.31?0.33 nmol/mg protein,\r\np=0.0001) in serum in thyrotoxic group versus control, while MDA levels did not differ\r\nsignificantly from the control. Significantly low values of the SH groups, GSH and SOD were\r\nfound (p<0.001) in the plasma of Thyroxin treated rats. Vitamin E supplementation\r\nsignificantly increased plasma MDA levels in the Thyroxin treated group as compared with\r\nthe control group (p=0.01) and with the animals treated only with Thyroxin (p=0.04).\r\nCarbonyl protein levels in plasma of the hyperthyroid supplemented rats were also increased\r\nas compared to controls (p=0.0002). Antioxidant capacity markers in plasma of group 3 were\r\ndecreased compared with group 1. The marker of lipid peroxidation (MDA) significantly\r\ndecreased in thyroid homogenates of the group 2 as compared with group 1 (p=0.004).\r\nSignificantly high levels of the SH groups (p=0.0006) and low levels of GSH (p=0.0001) were\r\nfound in thyroid homogenates of the L-Thyroxin treated group as compared with controls.\r\nThese results suggest that experimental hyperthyroidism is accompanied with\r\nincreased oxidative stress and with the consumption of antioxidant enzymes in induced\r\noxidative aggressions. No protective effects of vitamin E on oxidative stress induced by\r\nexcessive administration of thyroid hormones were detected.

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. In vitro studies of the changes about osteoblastogenesis and adipogenesis potential of BMSCs were not clear. As it is the critical pathway for osteogenic differentiation and bone formation, whether or not Wnt/β- catenin signalling is involved in the changes of osteogenic and adipogenic potential of BMSCs and participates in bone content decrease of ovariectomized (OVX)osteoporosis rats has been rarely reported. Material/Methods. BMSCs from femurs of ovariectomzed rats were isolated and cultured in vitro. The proliferation potential of BMSCs was analysed by CCK-8 assays . Osteoblastic and adipogenic differentiation potential of the BMSCs was assessed by ALP activity assay, Alizarin red S staining, Oil red O staining and RT-PCR analysis. Results. The results demonstrated that BMSCs from bilateral ovariectomization rats were endowed with lower proliferation and osteoblastic differentiation potential but higher adipogenic potential than the control group in vitro. In addition, β-catenin was found to have been decreased in OVX BMSCs, indicating that Wnt/β-catenin signalling pathways were suppressed in OVX BMSCs . Conclusions. Results suggested that changes in the Wnt canonical signalling pathway may be related to imbalances of osteogenic and adipogenic potential of BMSCs, and this may be an important factor related to bone content decrease in ovariectomized osteoporosis rats.

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.

Introduction: The impaired transport of leptin into the brain through a decreased permeability of the blood-brain barrier (BBB) for leptin in obesity represents one of the important mechanisms involved in leptin resistance which is characteristic in human obesity. Some pituitary tumors can increase the blood-cerebrospinal fluid barrier (BCB) permeability for peptides. BCB is a part of BBB.\r\nObjectives: The aim of our study was to search if the by-pass of BCB for pituitary hormones produced by adenomas does influence the transport of leptin into the central nervous system in obese patients.\r\nMaterials and methods: We investigated 20 males with pituitary adenomas: group A (11 patients) had cerebrospinal fluid (CSF) to serum ratio more than one for prolactin (PRL) and in some patients for growth hormone (GH) and follicle stimulating hormone (FSH), suggesting an increased permeability of BCB and a control group C (9 patients), which had CSF/serum ratio less than one for GH, PRL or FSH, suggesting an intact BCB. Both A and C groups contain subgroups of patients with obesity (body mass index, BMI>30 kg/m2) and normal body weight (BMI<25 kg/m2). In these patients we measured the CSF to serum leptin ratio in order to clinically evaluate the leptin transport into the brain. Rapid fluoroimmunoassay method with europium was used. Leptin was assayed by ELISA method.\r\nResults: The patients of group A with pituitary adenomas show a higher level of pituitary peptides, PRL and in some cases GH, FSH in CSF as compared to serum (ratio CSF/serum over 1), both in obese and non-obese. By contrast, in the same patients, there is\r\na low level of CSF leptin as compared to serum leptin (ratio CSF/serum less than 1). In the subgroup of obese patients from group A we found even less ratio of CSF to serum leptin, than in non-obese. There is a well known higher leptin concentration in the plasma of obese patients with pituitary adenomas as compared to non-obese ones (26.4?3.8ng/ml vs 12.4?3.4ng/ml, p<0.05). In the control group C, both pituitary peptides (PRL, or GH, FSH) and leptin showed a ratio CSF/serum less than 1, in all patients.\r\nConclusions: These data show a decrease in hemato-encephalic barrier permeability for leptin in obese patients through a specific mechanism, not influenced by other peptides passing through injuries of BBB produced by pituitary adenomas. It is tempting to suggest that there is a specific by-pass of BCB for pituitary peptides, in some pituitary adenomas.

Introduction. KCNJ11 gene activating mutations play a major role in the development of neonatal diabetes mellitus (NDM). KCNJ 11 gene encodes the Kir 6.2 subunit of ATP- sensitive potassium channel which is a critical regulator of pancreatic beta-cell insulin secretion. Aim. To study KCNJ11 gene mutations in infants with NDM and the effect of sulfonylurea treatment on the glycemic control in patients with KCNJ11 gene mutation. Subjects and methods. Thirty infants with NDM were screened for KCNJ11 gene mutations by DNA sequencing, insulin therapy was replaced by sulfonylurea treatment in patients with mutations. Results. R201C heterozygous mutation was found in one patient who was successfully shifted from insulin therapy to sulfonylurea treatment, while E23k, I337V, and S385C polymorphisms were detected in 14 patients. Conclusion. Screening for KCNJ 11 gene mutations could lead to identification of patients with mutations who can be successfully shifted from insulin therapy to sulfonylurea treatment improving their quality of life.