Anabolic Steroids-Driven Regulation of Porcine Ovarian Putative Stem Cells Favors the Onset of Their Neoplastic Transformation.

Nandrolone (Ndn) and boldenone (Bdn), the synthetic testosterone analogues with strong anabolic effects, despite being recognized as potentially carcinogenic compounds, are commonly abused by athletes and bodybuilders, which includes women, worldwide. This study tested the hypothesis that different doses of Ndn and Bdn can initiate neoplastic transformation of porcine ovarian putative stem cells (poPSCs). Immunomagnetically isolated poPSCs were expanded ex vivo in the presence of Ndn or Bdn, for 7 and 14 days.

Porcine ovarian cortex-derived putative stem cells can differentiate into endothelial cells in vitro.

Endothelial cells (ECs), the primary component of the vasculature, play a crucial role in neovascularization. However, the number of endogenous ECs is inadequate for both experimental purposes and clinical applications. Porcine ovarian putative stem cells (poPSCs), although not pluripotent, are characterized by great plasticity.

Efficient generation of neural-like cells from porcine ovarian putative stem cells - morphological characterization and evaluation of their electrophysiological properties.

Until recently, the mammalian ovary was considered to consist of fully differentiated tissues, but evidence for the presence of adult stem cells in this organ appeared. The differentiation potential of these cells, referred to as putative stem cells, is not well defined. Porcine ovarian putative stem cells (poPSCs) were immunomagnetically isolated from postnatal pig ovaries based on the presence of the SSEA-4 surface marker protein.

Proteolytically Degraded Alginate Hydrogels and Hydrophobic Microbioreactors for Porcine Oocyte Encapsulation.

In reproductive biology, the biotechnology revolution that began with artificial insemination and embryo transfer technology led to the development of assisted reproduction techniques such as oocyte in vitro maturation (IVM), in vitro fertilization (IVF) and cloning of domestic animals by nuclear transfer from somatic cell. IVM is the method particularly of significance. It is the platform technology for the supply of mature, good quality oocytes for applications such as reduction of the generation interval in commercially important or endangered species, research concerning in vitro human reproduction, and production of transgenic animals for cell therapies.

Effects of vinclozolin exposure on the expression and activity of SIRT1 and SIRT6 in the porcine ovary.

Within the mammalian reproductive system sirtuin 1 and 6 (SIRT1, SIRT6) are considered to contribute to steroid hormone signaling and control of reproductive physiology. Therefore, the specific question is whether and how a commonly used dicarboximide fungicide with antiandrogenic activity, vinclozolin (Vnz) alters SIRT1 and SIRT6 expression and whether both investigated sirtuins positively affect survival of the follicles after vinclozolin exposure. Immunocytochemistry and immunohistochemistry were performed to localize SIRT1 and SIRT6 expression in cultured granulosa cells (GCs; 48 hours) and whole ovarian follicles (24 hours) after treatment with two androgens, testosterone (T; 10 M) and dihydrotestosterone (DHT; 10 M), and an antiandrogen, Vnz (1.

Androgen receptor-mediated non-genomic effects of vinclozolin on porcine ovarian follicles and isolated granulosa cells: Vinclozolin and non-genomic effects in porcine ovarian follicles.

The present study investigated the influence of the androgen receptor (AR) agonists testosterone (T) and dihydrotestosterone (DHT), and vinclozolin (Vnz), a fungicide with antiandrogenic activity, on non-genomic signal transduction within ovarian follicles. Porcine granulosa cells (GCs) isolated from mature follicles were cultured for 48h. For the last 24h of culture, they were exposed to T (10(-7)M), DHT (10(-7)M), Vnz (1.

Comparison of Selected Morphological, Rheological and Biochemical Parameters of Winter Swimmers' Blood at the End of One Winter Swimming Season and at the Beginning of Another.

The aim of the study was to examine potential differences in the morphological, rheological and biochemical blood parameters of winter swimmers who remained physically active during the period between the end of one winter swimming season and the beginning of another. The study included a group of healthy winter swimmers (n = 17, all between 30 and 60 years of age). Six months following the end of winter season, the levels of mean corpuscular hemoglobin concentration and mean corpuscular hemoglobin turned out to be significantly higher, while erythrocyte count and hematocrit level significantly lower than at the baseline.

Effects of testosterone and prolactin on steroidogenesis in post-ovulatory cumuli oophori and on in vitro oocyte fertilisation in the rat.

The main objective of these studies was to determine the in vitro effects of prolactin (PRL) and testosterone (T) on steroidogenic function in post-ovulatory cumuli oophori containing unfertilised (ufCOCs) or fertilised (fCOCs) oocytes and to determine the differences between ufCOCs and fCOCs. In vivo, progesterone (P4) content was distinctly higher in isolated ampullae containing ufCOCs than in those containing fCOCs. Moreover, the expression of androgen (ARs) and prolactin (PRL-Rs) receptors was distinctly higher in ufCOCs than in fCOCs.

Analysis of porcine granulosa cell death signaling pathways induced by vinclozolin.

Recent studies suggest that disturbing androgen-signaling pathways in porcine ovarian follicles may cause granulosa cell (GC) death. For this reason, we investigated which apoptotic pathway is initiated after GC exposure to an environmental antiandrogen, vinclozolin (Vnz), in vitro. Immunocytochemistry, Western blots, and fluorometric assays were used to quantify caspase-3 and -9 expression and activity.

The effects of the environmental antiandrogen vinclozolin on the induction of granulosa cell apoptosis during follicular atresia in pigs.

The aim of this study was to investigate whether the androgens testosterone and dihydrotestosterone (DHT) and the antiandrogenic fungicide vinclozolin (Vnz) exert proapoptotic effects on porcine granulosa cells (GCs), and to examine the roles of these compounds in follicular atresia. Granulosa cells isolated from pig follicles were cultured for 24 hours, and then exposed to 0.1 μM testosterone, 0.

The impact of antiandrogen 2-hydroxyflutamide on the expression of steroidogenic enzymes in cultured porcine ovarian follicles.

We used our model system for agonism and antagonism of the androgen receptor (AR), in which the porcine ovarian follicles were exposed on the excessive concentration of an AR agonist- testosterone (T) or an AR antagonist- 2-hydroxyflutamide (2-Hf) to: (1) analyze the spatiotemporal expression of ovarian 3β-hydroxysteroid dehydrogenase (3β-HSD), cytochrome P450 17α-hydroxylase/c17,20-lyase (P450c17) and cytochrome P450 aromatase (P450arom); (2) to determine the contribution of AR-mediated action during steroidogenesis and (3) to establish some correlations between the onset and expression pattern of the investigated proteins. Whole follicles (6-8 mm in diameter) isolated from mature porcine ovaries have been incubated (for 24 h) in an organ culture system in the presence of T (10(-7 )M), 2-Hf (1.7 × 10(-4) M) or both T and 2-hydroxyflutamide (T+2-Hf, at the same concentrations as when added separately).

The influence of winter swimming on the rheological properties of blood.

The aim of this study was to analyze the changes in blood rheology resulting from regular winter swimming. The study was carried out on 12 male winter swimmers. Venous blood for morphological, biochemical and rheological analysis was sampled twice from each winter swimmer - at the beginning of the season and after its completion.

Effects of testosterone and 2-hydroxyflutamide on progesterone receptor expression in porcine ovarian follicles in vitro.

The purpose of the study was to test the possible role of the androgen receptor (AR) agonist (testosterone; T), an AR antagonist (2-hydroxyflutamide; 2-Hf) or combination of both (T+2-Hf) on progesterone receptor (PGR) expression in cultured porcine granulosa cells (GCs) or whole follicles. GCs isolated from mature pig follicles (6-8mm in diameter) were cultured for 48 h. Experimental cultures were carried out with the addition of T (10(-7)M), 2-Hf (1.

The influence of the antiandrogen 2-hydroxyflutamide on the androgen receptor expression in the porcine ovarian follicles - an in vitro study.

Androgens are one of the most important agents influencing ovarian follicles growth and development. The biological action of androgens is primarily exerted through transcriptional regulation by the androgen receptor (AR), a member of the steroid hormone receptor superfamily. The purpose of this study was to test the role of androgen receptor agonist testosterone (T) or antagonist 2-hydroxyflutamide (2-Hf) and in combination on AR expression in cultured porcine granulosa cells (GC) or whole follicles.

Steroid concentrations and immunoexpression of steroidogenic enzymes in ovaries of aged bank voles: effect of photoperiod.

The main objective of the present study was to establish morphological and steroidogenic changes occurring in the ovaries of senescent bank voles, with respect to the photoperiod of rearing. Obtained results revealed less pronounced changes in the ovaries of females reared in a long photoperiod (LD). Their gonads still possessed some healthy follicles and old corpora lutea (CLs).

Does 2-hydroxyflutamide inhibit apoptosis in porcine granulosa cells? - An in vitro study.

In mammalian ovaries, the majority of follicles are lost before ovulation by atresia. This degenerative process is initiated or caused by granulosa cell apoptosis. To reveal the androgen-dependent mechanism of selective follicular atresia, the culture model system for agonism and antagonism of the androgen receptor has been established.

The effects of exercise in water at 4°C and 25°C on the rheological properties of blood and the composition of fatty acids in the erythrocyte membranes of laboratory rats.

The aim of this study was to assess the influence of a single session of maximal exercise performed in water (4°C or 25°C) on blood rheological properties and the composition of fatty acids in the erythrocyte membranes of laboratory rats. This study will permit better understanding of the reactions occurring in the organism during rapid cooling in cold water, especially in regards to the hemorheological and biochemical parameters of blood. When compared to the control group, exercise performed in water at 4°C led to an increase in the elongation index (EI, from 0.

Effects of cold water swimming on blood rheological properties and composition of fatty acids in erythrocyte membranes of untrained older rats.

This is the first report on the effects of a single bout of swimming to exhaustion in cold water on rat erythrocyte deformability, aggregation and fatty acid composition in erythrocyte membranes. The results indicate that there was a significant decrease in body temperature of experimental rats swimming in water at 4 degrees C and 25 degrees C when compared to the control. Erythrocyte aggregation indices did not change after swimming in water at 4 degrees C whereas erythrocyte deformability increased at shear stress 1,13 [Pa] and 15,96 [Pa].

Luteal macrophage conditioned medium affects steroidogenesis in porcine granulosa cells.

The purpose of the study was to examine the effect of luteal macrophage conditioned medium (LMCM) on progesterone and estradiol production by cultured granulosa cells. Porcine granulosa cells were cultured for 48 h with or without LMCM in the absence or presence of 100 ng/ml LH, FSH or prolactin. Progesterone and estradiol concentrations were measured by radioimmunoassay.

Steroid levels and the spatiotemporal expression of steroidogenic enzymes and androgen receptor in developing ovaries of immature rats.

Immunoexpression of 3β-hydroxysteroid dehydrogenase (3β-HSD), cytochrome P450c17 (P450c17), androgen receptor (AR), and steroid contents were studied in the ovaries of immature female Wistar rats killed between postnatal days 1 and 30. During days 1-7, ovarian somatic structures lacked AR, 3β-HSD and P450c17, except for the surface epithelium, which featured the presence of these three proteins, suggestive of its androgen responsiveness and steroidogenic function. On day 10, AR appeared in many somatic structures, including the granulosa layers, which coincided with the P450c17 immunoexpression in some theca/interstitial cells, and an increase in ovarian androgen concentration.

Immunohistochemical study on differential distribution of progesterone receptor A and progesterone receptor B within the porcine ovary.

Progesterone plays a central role in the regulation of ovarian functions. Progesterone receptor (PGR) is expressed as two isoforms, PGRA and PGRB, which have been shown to have different functional activities. This immunohistochemical study describes the diverse localization of PGRA and PGRB in the porcine ovarian components: small and large antral follicles, early, midluteal and regressing corpora lutea.

Ectopic bone formation after treatment with colchicine.

We followed changes occurring within bone tissue and marrow cells during the process of colchicine-induced ectopic bone development and its resorption inside the marrow cavity of the rat tibia. To stimulate ectopic bone formation male Wistar rats were i.p injected with 0.

Development of rat tibia innervation: colocalization of autonomic nerve fiber markers with growth-associated protein 43.

Development of autonomic innervation of the tibia was investigated in rat fetuses on gestational days (GD) 17-21 and in juvenile animals on postnatal days (PD) 1-28. Double immunofluorescence combined with confocal microscopy was applied to study colocalization of neuronal growth- associated protein 43 (GAP-43) and panneuronal marker protein gene product 9.5 (PGP) with markers of the autonomic nervous system: neuropeptide Y (NPY) and dopamine beta-hydroxylase (DbetaH) for adrenergic, as well as vasoactive intestinal polypeptide (VIP) and vesicular acetylcholine transporter (VAChT) for cholinergic fibers.

Haematopoiesis in snakes (Ophidia) in early postnatal development.

The occurrence of haematopoiesis has been studied in various parts of the spine and in the ribs in four species of snakes (Boa constrictor L., Elaphe guttata L., Lamprophis fulaginosus Boie.

Androgen receptor in early apoptotic follicles in the porcine ovary at pregnancy.

Localization of androgen receptor (AR) was investigated in ovarian follicles developing and undergoing atresia during pregnancy in the pig. Immunohistochemical staining was conducted on ovarian antral follicles isolated on different days of gestation: 10, 18, 32, 50, 70, and 90. Paraffin sections were also subjected to in situ DNA labeling.