ACTA ENDOCRINOLOGICA (BUC)

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.

Testosterone influences cancer development. This in vitro experiment was exerted to determine the association of testosterone with human colorectal cancer(HT29), glioblastoma (A172) and human embryonic kidney(HEK293) cells proliferation. HT-29, A172 and HEK293 cell lines were cultured in standard growth medium, then randomly divided into control group (not exposed to testosterone) and groups exposed to 1, 10, 100 and 1000 μg/mL of testosterone. Cell viability was quantified by MTT assay. Statistical analysis was performed using ANOVA. Viability of HEK293 cells significantly increased in groups exposed to 1 μg/mL and decreased in groups exposed to 100 and 1000 μg/mL of testosterone compared to control group (P<0.05, P<0.05 and P<0.001, respectively). Viability of HT29 cells significantly increased in groups exposed to 10 and 100 μg/mL of testosterone and significantly decreased when exposed to 1000 μg/mL of testosterone compared to control group (P<0.05, P<0.001 and P<0.001, respectively). Viability of A172 cells significantly decreased in groups exposed to 100 and 1000 μg/mL of testosterone compared to control group (P<0.001). In conclusion, different doses of testosterone have enhancing or suppressive effects on HEK293, HT29 and A172 cells proliferation; according to which, considering clinical use of testosterone therapy for cancer treatment is a highly controversial issue.

Background. Resistin, as an adipokine, has been shown to be increased in serum plasma of gastric cancer patients and suggested to be a major factor in gastric carcinogenesis. However, it is still not clear how Resistin influences gastric cancer progression. The aim of this study was to evaluate Resistin effect on cell proliferation and expression of telomerase gene in gastric cancer cell line (AGS). Methods. In this study, the proliferating activity of AGS cells stimulated with Resistin was also evaluated by using 2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2HTetrazolium- 5-Carboxanilide (XTT) assay and trypan blue staining method. To investigate telomerase gene expression affected by Resistin, total RNA was extracted, cDNA was synthesized and expression of hTERT mRNA was carried out by real-time reverse transcription polymerase chain reaction. Results. Exogenous Resistin has induced gastric cancer cells proliferation in a dose-dependent manner and could improve cell viability. Also the expression of Human Telomerase Reverse Transcriptase (hTERT) was upregulated in 24 hours, after Resistin treatment. Conclusions. This study has shown Resistin induces exogenously gastric cancer cell proliferation and increases hTERT gene expression. These findings may clarify the role of Resistin in gastric carcinogenesis. Therefore blocking Resistin signaling and limiting its secretion may be valuable for the treatment of gastric cancer.

Leptin is much more than a simple sensor of body fat levels. Its biological activities, include angiogenesis, osteogenesis and modulation of the immune and reproductive systems. There are no reports on fluctuations of the level in normal human functional glandular epithelium during the menstrual cycle. Objective. We aimed to determine the expression level of the leptin receptor by immunohistochemistry and to correlate this with estrogen and progesterone receptor levels in normal endometrial tissue throughout the menstrual cycle. Study design. Thirty-seven paraffin wax blocks of biopsy specimens of histologically normal endometrium were evaluated and classified according to the stage of the menstrual cycle during the 2004–2006 period. Results. The OB-R level underwent fluctuations during the menstrual cycle in the different tissue compartments; these were most marked in the functional glandular epithelium and during the proliferative stage. OB-R levels were correlated positively with the expression levels of estrogen and negatively with those for progesterone receptors, suggesting that these steroids modulate the expression of OB-R in vivo. Conclusion. This finding underscores the importance of considering the menstrual cycle stage for studies seeking possible impacts of the OB-R on reproductive pathology, because its expression level varies with the hormonal environment.

The effects of daily evening melatonin (MT) injections on plasma T3 and T4 and pineal thyroxin 5&#8217;-deiodinase (5&#8217;-D) activity in euthyroid and hypothyroid rats were investigated. Circulating levels of thyroid hormones were monitored and 5&#8217;-D activity was measured in pineal homogenates throughout the daily light-dark cycle. In the euthyroid group, T3 and T4 concentrations and pineal 5&#8217;-D activity gradually increased during the L-phase of the L/D cycle to reach maximum levels early at night. The lowest values for pineal 5&#8217;-D activity and T4 were obtained later at night when endogenous MT production was the highest. MT treatment induced an opposite circadian variation of plasma T3, T4 and pineal 5&#8217;-D activity with significant increases later at night and decreases early at night vs. the control group. In the hypothyroid group, the serum T4 and T3 concentrations significantly decreased at all moments assayed. Treatment with MT did not lead to significant changes in the propylthiouracil effect on T4 and T3 levels, but maintained the biphasic response, observed in the MT treated euthyroid group. The increases induced by PTU in pineal 5&#8217;-D activity during the light phase, were reduced from 43.61 ? 2.35 ng T3/mg protein / h to 33.36?2.87 ngT3/mg protein/h (p=0.01) by MT injections. In conclusion, the results rendered the presence of the 5&#8217;-D in the rat pineal, its activity showing a circadian pattern similar to the circulating T4 levels. The MT treatment induced an opposite circadian variation of serum T3, T4 and pineal 5&#8217;-D activity suggesting an interaction between the light/dark cycle, 5&#8217;-D activity and responsiveness to MT.

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.

Background. Warthin-like tumor of the thyroid (WALTT) is a very rare variant of papillary thyroid cancer.We want to draw attention to this rare condition by reporting two cases. Patient reports. Patient 1 was a 24 year-old woman presented with 14×12 mm solid nodule on the left lobe of the thyroid gland. Fine needle aspiration biopsy of the nodule was reported as suspicious for papillary thyroid carcinoma. Patient 2 was a 40 year-old woman who had multinodular thyroid gland with a 31×26 mm major nodule on the left lobe. On fine neddle aspiration, cytologic findings were consistent with papillary thyroid carcinoma. Both underwent total thyroidectomy and histological examination of the cases revealed Warthin-like tumor of the thyroid. Summary and conclusion. We report two patients with WALTT. This rare variant of papillary thyroid cancinoma has four main histologic criteria: papillary architecture, oxyphilic cytoplasmic changes, papillary nuclear features and dense lymphoid infiltrate. WALTT can be distinguished from other aggressive variants with these distinct histological features. Since variants show different clinical behaviour, classification of these might be helpful to predict patient prognosis.

The aim of this study is to investigate the possible association between hormonal status and adipose tissue characteristics in obese and non-obese subjects. Fourteen obese and 15 nonobese premenopausal female patients were enrolled in the study. Stromal vascular cells were isolated and cultured using modified procedures described by Entenmann and Hauner. In the non-obese group, omental SVCs seeded at a density of 4.12?1.1x103/cm2 in 25-cm2 in culture flasks for measuring cell proliferation and subcutaneus SVCs seeded at a density of 2.05?0.76x103/cm2 in 25-cm2 at culture flasks. In the obese group, omental SVCs seeded at a density of 6.11?1.98x103/cm2 in 25-cm2 at culture flasks for measuring cell proliferation and subcutaneous SVCs seeded at a density of 2.94?0.75x103/cm2 in 25-cm2 in culture flasks. Mean GPDH activity levels were significantly higher in SVCs from the omentum in obese compared to those from the omentum in nonobese (651.9?65.7 vs 405.1?60.1 mU/mg protein). However, GPDH activities were similar in SVCs from the subcutaneous SVCs in obese subjects, compared to those from the subcutaneous SVCs in non-obese subjects (303.5?63.2 vs 367.4?73.7 mU/mg protein). In obese group, omental SVCs number was positively correlated with plasma estradiol (E2) (r=0.604, p=0.017), and fasting insulin levels (r=0.843, p=0.01). It was negatively correlated with plasma progesterone (r=-0.793, p=0.006), prolactin (r=-0.655, p=0.008) and free T3 (FT3) levels(r=-0.630, p=0.01). These findings suggest that there are differences in adipose tissue proliferation capacity and metabolic activity between obese and non-obese subjects. In obese group, the number of omental stromal vascular cells was positively correlated with plasma estradiol and insulin levels.

Context. Heat Shock Protein 60 (HSP60) is a chaperone protein which is involved in proteins transfer and re-folding of proteins. Objective. Importance of HSP60 in sperm capacitation and facility of sperm-oocyte membrane binding was confirmed, therefore in this study the effect of HSP60 on the rate of in vitro fertilization and the cleavage rate in mouse embryo was investigated. Design. Ten male mice and twenty five female mice were involved to collect sperms and oocytes required for this study. Subjects and Methods. Sperms were collected from the epididymis of male mouse and oocytes were collected from the oviduct of female mouse following ovarian hyperstimulation. Then, capacitated sperms and oocytes were placed together in fertilization medium in four groups in the presence of different concentrations of HSP60 (10, 50 and 100 ng/mL) and in the absence of HSP60. After calculation of the fertilization rate, zygotes were transformed into the other medium for development and the cleavage rate was monitored to blastocyst stage. Results. There was not a significant difference in the rate of fertilization between 10 ng/mL HSP60 group and the control group. The rate of fertilization and two-cell embryo development decreased significantly (P≤0.05) in 100 ng/mL HSP60 compared to other experimental and control groups. Further, the rate of two-cell embryo development increased significantly (P≤0.05) in 10 ng/mL HSP60 compared to other experimental and control groups. Conclusions. The present study demonstrated that HSP60 in low dose had a positive effect on two-cell embryo development, however it did not have any significant effect on the fertilization rate. Conversely, HSP60 had adverse effects on the fertilization and cleavage rates at higher doses.

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.