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Various animal models are used in the study of alcoholism, with the honeybee (Apis mellifera L.) among them. Here, we tested the hypothesis that foragers show higher intoxication resistance to alcohol than nurses, an issue thus far not investigated. To this end, we measured the latency to full sedation when exposed to alcohol in foragers, nurses and reverted nurses. In addition, we measured alcohol dehydrogenase (ADH) levels in these worker castes. Caste status was confirmed by comparison of the size of their hypopharyngeal glands. We detected high intoxication resistance to alcohol and presence of ADH in foragers. In nurses, we detected significantly lower intoxication resistance to alcohol and no ADH. These between-caste differences cannot be explained by the age difference between castes as in reverted nurses, characterized by similar age to foragers, we detected an intermediate intoxication resistance to alcohol and no ADH. Our results suggest possible natural exposure to alcohol in different castes of workers. As such, we further develop the honeybee as a model in alcoholism-related research and open new research avenues.

BACKGROUND Notch signaling pathway is involved in contact-dependent communication between the cells of seminiferous epithelium, and its proper activity is important for undisturbed spermatogenesis. OBJECTIVES The aim was to assess the effect of Notch pathway inhibition on the expression of nuclear (AR) and membrane (ZIP9) androgen receptors and androgen-regulated genes, claudin-5 and claudin-11, in TM4 mouse Sertoli cell line. MATERIALS AND METHODS DAPT (γ-secretase inhibitor) treatment and recombination signal binding protein silencing were employed to reduce Notch signaling, whereas immobilized ligands were used to activate Notch pathway in TM4 cells. To reveal specific effect of each androgen receptor, AR or ZIP9 silencing was performed. RESULTS Notch pathway inhibition increased the expression of AR and ZIP9 mRNA and proteins (p

Our present knowledge on interrelation between morphology/ultrastructure of mitochondria of the Leydig cell and its steroidogenic function is far from satisfactory and needs additional studies. Here, we analyzed the effects of blockade of androgen receptor, triggered by exposure to flutamide, on the expression of steroidogenic proteins (1) and ultrastructure of Leydig cells’ constituents (2). We demonstrated that increase in the expression level of steroidogenic (StAR, CYP11A1, 3β-HSD, and CYP19A1) proteins (and respective mRNAs) in rat testicular tissue as well as elevation of intratesticular sex steroid hormone (testosterone and estradiol) levels observed in treated animals correspond well to morphological alterations of the Leydig cell ultrastructure. Most importantly, up-regulation of steroidogenic proteins’ expression apparently correlates with considerable multiplication of Leydig cell mitochondria and subsequent formation of local mitochondrial networks. Interestingly, we showed also that the above-mentioned processes were associated with elevated transcription of Drp1 and Mfn2 genes, encoding proteins implicated in mitochondrial dynamics. Collectively, our studies emphasize the importance of mitochondrial homeostasis to the steroidogenic function of Leydig cells.

Background: A review of epidemiological data demonstrates relationships between defined health behaviours linked with religious affiliation and a reduced risk of developing and dying from Cardiovascular Disease (CVD). The aim of the study was to compare the lifestyle of Seventh-day Adventists (SDA) and Catholics (CA), to determine the relationship between the lifestyle of SDA, CA and the level of serum homocysteine as a risk factor for CVD. Methods: A cross-sectional study was conducted on 252 SDA and CA. The following tools were used: interview questionnaire, anthropometric measurement, the International Physical Activity Questionnaire (IPAQ), the Inventory of Health Behaviours (IHB), the Perceived Stress Scale (PSS-10), laboratory tests (homocysteine level), and the Fagerstr&ouml, m Test for Nicotine Dependence (FTND). Results: Selected lifestyle elements, such as smoking cigarettes, drinking alcohol, physical activity, diet, Body Mass Index (BMI), health behaviours on the IHB, psychosocial factors and level of stress for CA were significantly different in comparison to SDA. The religion professed by the respondents was not significantly associated with the increased level of homocysteine as a risk factor for cardiovascular diseases (CVD). The level of homocysteine for CA were significantly different in comparison to SDA. The studied group of CA was significantly influenced by socio-demographic factors: gender, age, education, place of residence, BMI and lifestyle: drinking alcohol and smoking cigarettes, consumption of dark bread, pasta, and groats. For the studied group of SDA, the level of homocysteine was significantly influenced by socio-demographic factors such as gender, age, professional activity, and consumption of legumes. Conclusions: Public health professionals and nurses should develop culturally specific educational interventions.

Background Onset of spermatogenesis at puberty is critically dependent on the activity of hypothalamic-pituitary-gonadal axis and testosterone production by Leydig cells. The aim of this study was to examine whether activation of Notch receptors and expression of Notch ligands and effector genes in rat seminiferous epithelium are controlled by androgen signaling during puberty. Methods Peripubertal (5-week-old) Wistar rats received injections of flutamide (50 mg/kg bw) daily for 7 days to reduce androgen receptor (AR) signaling or a single injection of ethanedimethane sulphonate (EDS; 75 mg/kg bw) to reduce testosterone production. Gene and protein expressions were analyzed by real-time RT-PCR and western blotting, respectively, protein distribution by immunohistochemistry, and steroid hormone concentrations by enzyme-linked immunosorbent assay. Statistical analyses were performed using one-way ANOVA followed by Tukey’s post hoc test or by Kruskal-Wallis test, followed by Dunn’s test. Results In both experimental models changes of a similar nature in the expression of Notch pathway components were found. Androgen deprivation caused the reduction of mRNA and protein expression of DLL4 ligand, activated forms of Notch1 and Notch2 receptors and HES1 and HEY1 effector genes (p p p p p p Conclusions Androgens and androgen receptor signaling may be considered as factors regulating Notch pathway activity and the expression of Hes and Hey genes in rat seminiferous epithelium during pubertal development. Further studies should focus on functional significance of androgen-Notch signaling cross-talk in the initiation and maintenance of spermatogenesis.

Adipokines influence energy metabolism and have effects on male reproduction, including spermatogenesis and/or Sertoli cell maturation, however, the relationship between these active proteins and androgens in testicular cells is limited. Here, we studied the impact of short-term exposure to flutamide (an anti-androgen that blocks androgen receptors) on the expression of chemerin, apelin, vaspin and their receptors (CCRL2, CMKLR1, GPR1, APLNR, GRP78, respectively) in adult rat testes. Moreover, the levels of expression of lipid metabolism-modulating proteins (PLIN1, perilipin1, TSPO, translocator protein) and intercellular adherens junction proteins (nectin-2 and afadin) were determined in testicular cells. Plasma levels of adipokines, testosterone and cholesterol were also evaluated. Gene expression techniques used included the quantitative real-time polymerase chain reaction (qRT-PCR), Western blot (WB) and immunohistochemistry (IHC). The androgen-mediated effects observed post-flutamide treatment were found at the gonadal level as chemerin, apelin, and vaspin gene expression alterations at mRNA and protein levels were detected, whereas the cellular targets for these adipokines were recognised by localisation of respective receptors in testicular cells. Plasma concentrations of all adipokines were unchanged, whereas plasma cholesterol content and testosterone level increased after flutamide exposure. Differential distribution of adipokine receptors indicates potential para- or autocrine action of the adipokines within the rat testes. Additionally, changes in the expression of PLIN1 and TSPO, involved in the initial step of testosterone synthesis in Leydig cells, suggest that testicular cells represent a target of flutamide action. Increase in the gene expression of PLIN1 and TSPO and higher total plasma cholesterol content indicates enhanced availability of cholesterol in Leydig cells as a result of androgen-mediated effects of flutamide. Alterations in adherens junction protein expression in the testis confirm the flutamide efficacy in disruption of androgen signalling and presumably lead to impaired para- and autocrine communication, important for proper functioning of adipokines.

W kręgu zainteresowań specjalistów zdrowia publicznego i pielęgniarek jest identyfikacja tych elementów stylu życia, które zapobiegają chorobom w określonych populacjach ludzkich, w tym w grupach religijnych. Określenie związku stylu życia katolików ze stężeniem homocysteiny w surowicy krwi, jako czynnika ryzyka chorób układu krążenia (CVD). Badania przekrojowe przeprowadzono wśród 134 katolików. Wykorzystano kwestionariusz ankiety własnego autorstwa, Międzynarodowy Kwestionariusz Aktywności Fizycznej, Inwentarz Zachowań Zdrowotnych (IZZ), Skalę Odczuwanego Stresu PSS-10, badania laboratoryjne (stężenie homocysteiny), Test Uzależnienia od Nikotyny Fagerströma. U ponad połowy badanych katolików zdiagnozowano podwyższone stężenie homocysteiny. Wiedza niespełna połowy badanych na temat CVD była w pełni zadowalająca. Wybrane elementy stylu życia katolików, jak aktywność fizyczna, nałogi, odczuwany stres, zachowania zdrowotne w IZZ, czynniki psychospołeczne oraz dieta odbiegały od zaleceń istotnych w prewencji CVD i nie były istotnie powiązane z wynikami stężenia homocysteiny w surowicy krwi, z wyjątkiem istotnej zależności pomiędzy jej prawidłowym poziomem a spożywaniem mleka, płatków wielozbożowych, makaronów i kasz pszennych oraz niepaleniem papierosów. Specjaliści w zakresie zdrowia publicznego i pielęgniarki powinni rozwijać specyficzne kulturowo interwencje edukacyjne, uwzględniające styl życia i stężenie homocysteiny, jako czynniki ryzyka chorób układu krążenia wśród wyznawców różnych religii. Public health specialists and nurses are interested in identifying the elements of lifestyle that prevent disease in specific human populations, including religious groups. The term lifestyle catholic compound with the level of homocysteine in blood serum, as a risk factor for cardiovascular disease (CVD). The cross-sectional study was conducted among 134 catholics. Use of a questionnaire of own authorship, the International Physical Activity Questionnaire, Inventory Behavioral Health (HBI), Scale Experienced Stress PSS-10, laboratory tests (homocysteine), Test Fagerström Nicotine Dependence. In more than half of catholics diagnosed increased homocysteine levels. Knowledge of about halves of the respondents on CVD was fully satisfactory. Selected elements of life style catholics, like physical activity, addictions, perceived stress, health behaviors in the HBI, psychosocial factors and diet deviate from the recommendations relevant to the prevention of CVD and were not significantly linked to the performance level of homocysteine in the blood serum, with the exception of a significant relationship between its proper level and the consumption of milk, whole wheat flakes, pasta and wheat groats and also not smoking cigarettes. Public health professionals and nurses should develop culturally specific educational interventions, taking into the life style and homocysteine level account cardiovascular risk factors among followers of different religions.

Leydig cell tumors (LCT) are the most common type of testicular sex cord-stromal tumor. In this report, we implicate the G-coupled estrogen receptor (GPER) and peroxisome proliferator receptor (PPAR) in regulation of lipid homeostasis and the expression of steroidogenesis-controlling molecules in clinical specimens of LCTs and cell line (mouse tumor Leydig cells; MA-10). We also show the general structure and morphology of human LCTs with the use of scanning electron microscopy and light microscopy, respectively. In LCTs, protein immunoblotting and immunohistochemical analysis revealed increased expression of GPER and decreased expression of PPARα, β and γ. Concomitantly, changes in expression pattern of the lutropin receptor (LHR), protein kinase A (PKA), perilipin (PLIN), hormone sensitive lipase (HSL), steroidogenic acute regulatory protein (StAR), translocator protein (TSPO), HMG-CoA synthase (HMGCA), and HMG-CoA reductase (HMGCR) were observed.Using MA-10 cells treated with GPER and PPAR antagonists (alone and in combination), we demonstrated there is a GPER-PPAR mediated control of cholesterol concentration. In addition, GPER-PPARα regulated estradiol secretion, while GPER-PPARγ affected cGMP concentration. It is assumed that GPER and PPAR can be altered in LCT, resulting in a perturbed lipid balance and steroidogenesis. In LCTs, the phosphatidylinositol-3-kinase (PI3K)-Akt-mTOR signaling pathway was disturbed. Thus, PI3K-Akt-mTOR, together with cGMP, can play a role in LCT proliferation, growth, and metastasis as well as lipid balance control.In conclusion, we discuss the implications of GPER-PPAR interaction with lipid metabolism and steroidogenesis controlling-molecules in LCT biology that can be used in future studies as potential targets of diagnostic and therapeutic implementations.

The study was designed to examine the effects of model plastic derived compounds, bisphenol A (BPA) and dibutyl phthalate (DBP), on juxtacrine communication in adult rat testis, by evaluating the expression of Notch pathway components. Testicular explant were exposed in vitro to BPA (5 × 10−6 M, 2.5 × 10−5 M, 5 × 10−5 M) or DBP (10−6 M, 10−5 M, 10−4 M) for 24 h. To determine the expression of Notch1, Dll4, Hey1, Hes1 and Hey5 real-time RT-PCR was used. Protein levels and localization of NOTCH1 receptor, its ligand DLL4 as well as HEY1, HES1 and HEY5 factors were detected by western blot analysis and immunohistochemistry, respectively. Upregulation of Notch1, Dll4 and Hey1 at the mRNA and protein level was demonstrated in testis explants after BPA and DBP treatment (p

This study aimed to investigate rapid effect of anti-androgen 2-hydroxyflutamide (HF) on cadherin/catenin complex and androgen receptor (AR) phosphorylation in prostate cancer cell lines. In addition, a role of PI3K/Akt and MAPK/ERK1/2 pathways in mediating these effects was explored. We have demonstrated that in androgen-sensitive LNCaP cells HF induced rapid increase of E-cadherin phosphorylation at Ser 838/840 (p0.05) in MAPK/ERK1/2-dependent manner, whereas phosphorylation of β-catenin at Tyr 654 was unchanged. Concomitantly, the reduction of the level of AR phosphorylated at Ser210/213 was found (p0.01). In androgen-independent PC3 cells HF decreased Tyr 860 N-cadherin and Tyr 645 β-catenin phosphorylation (p0.01), acting via both MAPK/ERK1/2 and PI3K/Akt pathways. Further, we evidenced that MAPK/ERK1/2 and PI3K/Akt pathways were differentially influenced by HF in LNCaP and PC3 cells. In LNCaP cells, both Akt (p0.01) and ERK1/2 (p0.001) phosphorylation were negatively regulated and this effect was mediated by Raf-1 (p0.05). In contrast, in PC3 cells HF stimulated Akt (p0.001) and ERK1/2 (p0.001) activation, but had no effect on the crosstalk between PI3K/Akt and MEK/ERK1/2 pathways at the Raf-1 kinase level. Our findings expand the role of anti-androgen into non-genomic signaling, creating a link between anti-androgen action and phosphorylation of adherens junction proteins in prostate cancer cells.

We report the effects of polychlorinated napthalanes (PCNs) on the mRNA expression of estrogen-related receptors (ERRs) α, β and γ, calcium (Ca2 + ) concentration, and sex steroid secretion in mouse primary and tumor Leydig cells. The cells were exposed to a mixture of PCNs (10 nM) alone or in combination with one of sex steroid receptor antagonists; 182,780 (ICI; 10 μM); hydroxyflutamide (HF; 10 −4 M) and G-coupled estrogen receptor antagonist (G15; 10 nM) respectively. The expression of mRNAs and protein for ERRα, β, and γ was detected in primary and tumor Leydig cells. The expression of ERRs was always lower in primary Leydig cells. Exposure of Leydig cells to PCNs significantly increased the expression of ERRs mRNA irrespective of the cell type. Concomitantly, an increased concentration of Ca2+ and sex steroids was revealed in exposed cells. After ICI, HF or G15 was added no changes in expression of ERRs was found. In Leydig cells changes in ERRs expression at mRNA level are clearly linked to changes in Ca2+ level and steroid secretion. Estrogen and androgen receptors are not involved in PCNs action in Leydig cells. The effect of PCNs on mouse Leydig cells is independent on the cell of origin (primary or tumor).