It is well documented that pesticides used in agricultural processes may have detrimental effects upon human health. Moreover, many of these compounds have been indicated as potential endocrine and reproductive disruptors. In the present study, the ability of herbicide acetochlor to affect a growth of estrogen-sensitive MCF-7 mammary epithelial carcinoma cells was studied using E-screen test. Acetochlor alone did not affect proliferation of MCF-7 cells. Significant inhibition of estradiol-induced (10-14-10-8 M) MCF-7 cell growth by the action of acetochlor (10-5 M) and interaction between these chemicals were observed. Estradiol-stimulated MCF-7 cell proliferation was decreased by 41% of positive control (17β-estradiol, 10-8 M, 100%) in the presence of acetochlor (10-5 M). Our results demonstrate that acetochlor might interfere with estradiol signaling conducted to altered proliferation activity of MCF-7 cells and might support endocrine disruptive effects of acetochlor.

The hypothalamic paraventricular nucleus is a key structure in the regulation of the autonomic and neuroendocrine systems response to acute and chronic stress challenges. In this study, we examined the effect of a mechanical posterolateral deafferentation of the PVN on the activity of sympathoadrenal system (SAS) and hypothalamo-pituitary-adrenal (HPA) axis by measuring plasma concentrations of epinephrine (EPI), norepinephrine (NE), and corticosterone (CORT) in rats exposed to acute immobilization (IMO) stress. The surgical posterolateral deafferentation of the PVN (PVN-deaf) was performed by Halasz knife, in brain of the adult male Sprague Dawley rats, according to coordinates of a stereotaxic atlas. Sham-operated (SHAM) animals underwent a craniotomy only. The animals were allowed to recover 14 days. Thereafter, the tail artery was cannulated and the animals exposed to acute IMO for 2 h. The blood samples were collected via cannula at the time points of 0, 5, 30, 60, and 120 min of the IMO. Concentrations of plasma EPI, NE, and CORT were determined by radioimmunoassay. The IMO-induced elevation of plasma EPI concentrations in the PVN-deaf rats reached statistical significance at 60 min of the IMO, when compared to SHAM rats. Similarly, the stress-induced elevation of the NE plasma levels in the PVN-deaf rats was significantly exaggerated at all time intervals of IMO in comparison with SHAM rats, whereas plasma CORT levels were significantly reduced. In contrast to the traditional view of excitatory role of the PVN in response to stress, our data indicate that some projections from the PVN to caudally localized hypothalamic structures, the brainstem or the spinal cord, exert inhibitory effect on the SAS system activity during acute IMO stress. The data indicate that stress-induced activation of the HPA axis is partially dependent on inputs from the brainstem to the PVN.

Effect of estrogenic compounds and 17β-estradiol (E2), which induces endothelial cell motility, was investigated on ROCK-2 expression in rat coronary vascular endothelial cells (CVEC). The CVEC were isolated from the heart of Wistar rats by collagenase (0.04%) and incubated with E2 (1-100 nM), estrogen receptor α (ERα) agonist: propyl pyrazole triol (PPT, 10 nM); ERβ agonists: (2,3-bis(4-hydroxyphenyl)-propionitrile, DPN, 10 nM) and E2-conjugate with bovine serum albumin (E2-BSA, 1 nM); and GPER1 agonist: G1 (100 nM). Furthermore, the effect of combination of E2 with estrogen receptors (ERs) antagonist and GPER1 agonist, ICI-182780 (10 µM), physiological estrogen antagonists: progesterone (P4, 10-100 nM) and testosterone (T, 10-100 nM); transcription inhibitor: actinomycin-D (1 µg/ml); GPER1 antagonist: G-15 (100 nM), superoxide dismutase, (SOD, 500 U/ml); Gi/o protein inhibitor: pertussis toxin (PTX, 100 µg/ml); and epidermal growth factor receptor (EGFR) blocker: AG-1478 (10 µM) was tested. After 24h incubation, ROCK-2 and GPER1 protein expressions were detected in the CVEC by Western-blotting. E2, ICI-182780, and G1 but not E2-BSA significantly up-regulated ROCK-2 expression, which was suppressed by actinomycin-D, PTX, AG-1478, and G-15. However, PPT and DPN had no effects on the ROCK-2 expression. ICI-182780, P4, T or SOD did not antagonize the E2 action. GPER1 expression was demonstrated in the CVEC. Estrogens could up-regulate ROCK-2 in the rat CVEC through GPER1 and EGFR transactivation.

The aim of the present study was to compare the prevalence of metabolic syndrome (MS) according to six definitions and evaluate the agreement between them in an urban sample in Qazvin, Iran. In a cross sectional study, 529 men and 578 women in age of 20-78 years were attended from September 2010 to April 2011 in Qazvin, Iran. Standardized measurements were available for waist circumference, blood pressure, fasting serum high-density lipoprotein cholesterol, triglycerides, glucose, and insulin. The diagnostic criteria proposed by World Health Organization (WHO), National cholesterol education program third Adult Treatment Panel (ATPIII 2001, 2004), International Diabetes Federation (IDF), American Heart Association/National Heart, Lung and Blood Institute (AHA/NHLBI), and new Joint Interim Societies (JIS) were applied. The degree of agreement between different definitions was assessed by kappa statistic. The prevalence of MS was found to be 28% (WHO), 26.2% (ATPIII 2001), 30.6% (ATPIII 2004), 34.2% (IDF), 33% (AHA/NHLBI), and 39.3% (JIS). With regards to the gender, the prevalence of MS according to ATPIII 2001 and IDF was significantly greater in women than men. However, using other definitions, the prevalence of MS was similar in both genders. The agreement of JIS criteria with IDF and AHA/NHLBI definitions was excellent (IDF: 0.902; AHA/NHLBI: 0.862), while the agreement of IDF criteria with ATPIII 2004 and AHA/NHLBI was good (ATPIII 2004:0.719; AHA/NHLBI: 0.756). Although JIS definition resulted in a higher prevalence of MS than other definitions, an excellent agreement has been seen between these criteria. Widespread acceptance of JIS definition may lead to management and therapy in a greater part of the population.

Bisphenol A (BPA) with its estrogenic properties is intensively studied since its presence in the environment and human body. Besides other adverse effects, the compound is suspected of contributing to hormone-related cancers. The present study was aimed to investigate short time (24 h) effects of BPA on the important genes/proteins involved in apoptosis and the cell cycle progression in the breast cancer cells MCF7. The experimental design covered cell treatment with a broad BPA concentration scale: 10-12M corresponding to ubiquitous exposure, 10-9M relevant to human levels, and 10-6M as experimentally usual. We further investigated the combined effects of low BPA dose (10-12M) with physiological concentration of estradiol (E2) (10-9M). The expression of particular proteins and genes was studied by Western blotting and real time RT-PCR, respectively. Estrogenic effect of BPA was confirmed in the following checkpoints: mRNA expression of estrogen receptor α, expression of cyclin D1 and A2 proteins and CCNA2 gene, Bax and Bcl2 protein levels. For both cyclins protein levels, the maximum stimulation was present at 10-9M BPA and the effects resembled the "inverted U"-shape, a nonmonotonic dose-response curve reported for the action of xenoestrogens. The combined effect of low BPA dose with physiological E2 concentration differ from those of individual compounds, the character of stimulatory response is neither additive nor synergistic. The results obtained strongly support the evidence of BPA and BPA+E2 proliferation-promoting effects in human breast carcinoma cells, even after short time exposure, partially via reduced rate of apoptosis by the action of BPA+E2.

The essential role of vitamin D (VD) in bone metabolism and mineral homeostasis is well established knowledge. Research indicates that classical and non-classical pathways of VD affect also cell proliferation and differentiation, the immune system, infection, and cancer. VD receptor (VDR) and VD metabolizing enzymes have been detected in female reproductive tissues, such as ovary, uterus and placenta. The presence of VD metabolites was demonstrated in follicular fluid (FF) in women undergoing in vitro fertilization and embryo transfer (IVF-ET). The recent studies show that VD regulates the expression of a large number of genes in reproductive tissues implicating a role for VD in female reproduction and pregnancy outcomes. There is increasing human and animal data suggesting that VD status may be associated with impaired fertility, endometriosis, polycystic ovary syndrome (PCOS), and ovarian cancer. The presence of VDR in both animal and human ovarian tissue has raised the question of a possible direct role for 1α,25-dihydroxyvitamin D [1α,25(OH)2D3] in the regulation of steroid hormone synthesis and secretion. Our recent data have demonstrated that 1α,25(OH)2D3 may affect in vitro insulin- and follicle-stimulating hormone (FSH)-induced progesterone secretion by porcine ovarian granulosa cells. The molecular mechanisms of this action should be further investigated.

In the everyday life, stress is deemed as something unfavorable that may enhance the risk for the development or worsen a disease. However, in its nature, stress is adaptive reaction of the body. Its main characteristic is the activation of the hypothalamic-pituitary-adrenocortical (HPA) axis. Previously, we have shown that activation of the HPA axis plays a gastroprotective role during acute stress. The aim of our study was to clarify the effects of chronic stress and chronically elevated basal corticosterone levels on the gastric ulceration and cardiovascular vulnerability in rats. Male Wistar rats were repeatedly restrained 60 min daily for 14 days and examined on day 15th. The gastric ulceration was induced by a s.c. injection of indomethacin (35 mg/kg). The cardiovascular vulnerability was examined in urethane-anaesthetized rats in an experimental angina pectoris model (epinephrine, 10 µg/kg, 30 s later phentolamine, 15 mg/kg, both i.v.). We confirmed the development of chronic stress consequences by changes in several somatic parameters (body weight decrease, thymus involution, adrenal gland hypertrophy), and elevated resting corticosterone levels. However, the gastroprotective effect of chronic stress was not manifested and there was no aggravation of indomethacin-induced gastric ulceration, either. In the experimental angina pectoris model, previous chronic stress did not have any profound effect on the blood pressure, heart rate, and electrocardiogram changes. In contrast to the general view on the harmfulness nature of the stress, we were unable to find a harmful effect of chronic stress on the internal diseases (gastric ulceration and angina pectoris). However, its protective effect was also missing among present experimental conditions.

It was aimed to investigate the effect of standardized ginseng extract on fertility parameters in diabetic rats. Thirty male rats were randomly allocated into three groups of 10 rats each: 1. controls, 2. diabetes (D) and 3. diabetes + ginseng (DG). The latter two groups were rendered diabetic by i.p. injection of streptozotocin (STZ; 50 mg/kg). Standardized ginseng extract (Dansk Droge A/S, Copenhagen, Denmark) was administered per os (100 mg/kg BW) by stomach tube daily for 90 days starting one week after STZ. Ninety days post STZ the rats were sacrificed, and testis, epididymis, prostate, and seminal vesicles were weighed and subjected to histological examination. In addition, spermiogram, testicular enzyme markers, intratesticular steroid hormonal profile and testicular antioxidant status were estimated. The administration of ginseng extract resulted in a significant improvement of fertility parameters and testicular antioxidants together with a decrease in malondialdehyde and testicular pathological signs including degenerative changes of the seminiferous tubules. Ginseng extract may be a beneficial adjuvant therapy for diabetics suffering from infertility as a complication.

The aim of the study is to evaluate the changes of bone mineral density (BMD), incidence of pathological fractures and to asses the effect of bisphosphonate therapy in prostate cancer patients (PCa) on androgen deprivation therapy (ADT) with the use of LHRH. In this prospective study bone mass density (BMD) was assessed by dual x-ray absorptiometry (DXA) in 97 PCa patients and 89 patients of compared group. DXA was examined at baseline and patients in the study group were subjected to ADT. PCa patients with osteoporosis were treated by calcium, vitamin D, and bisphosphonate and the subsequent DXA was made after 10 months. All other PCa patients (non-osteoporotic) had DXA examined every 12-14 months. Patients of the study group had significantly lower baseline L1-L4 and total hip BMD (p=0.028, p=0.022). BMD was significantly lower in L1-L4 and total hip (p=0.004, p<0.001, resp.) after 10-14 months and in L1-L4, femoral neck, and total hip (p=0.001, p=0.037, p< 0.001, resp.) after 20-26 months of ADT. After the treatment for osteoporosis with bisphosphonate a significant increase of BMD (p=0.04) was found in a total of 23 patients. Overall, the incidence of fractures after 20-26 months of ADT was 8.5 %. Osteopenia is very common in hormone naive PCa patients. There was a significant loss of BMD after 12 months of ADT which was progressive while the patients were on ADT. Bisphosphonate therapy was effective after 20 months of treatment. The incidence of pathological fractures was 7-fold higher in the study group.

The objective of this study was the detection of circulating tumor cells (CTCs) in metastatic breast cancer patients. Since only small numbers of circulating tumor cells (CTCs) are found in peripheral blood, at first we performed immunomagnetic separation as a concentration method suitable for selecting circulating tumor cells in peripheral blood. This was followed by analysis of isolated cells with the aid of laser scanning cytometry (LSC). Twenty eight patients with metastatic breast cancer were enrolled in the study and the control group consisted of 19 clinically healthy women. Six milliliters of peripheral blood was drawn for the analyses, but only in two patients the blood has been drawn twice. Blood samples were taken when no chemotherapy was administered, but hormonal therapy has been allowed. The positivity for CTCs was found in 20 (50.0 %) patients with metastatic breast cancer patients, while in 6 (31.6 %) healthy controls false positive circulating epithelial-like cells were detected. Because we did not use CD45 staining, we could not distinguish these circulating epithelial-like cells from CTCs. In a majority of metastatic breast cancer patients we found a mixed population of HER-2 gene expressing CTCs. We found that HER2+ CTCs in high numbers are CK19 + CTCs, while almost all HER2-CTCs are CK19- CTCs. The described method was found promising for estimating HER2 status on CTCs from peripheral blood in metastatic breast cancer patients.