Animal Production Research Center Nitra


Animal Production Research Center Nitra

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Sirotkin A.V.,Constantine the Philosopher University | Sirotkin A.V.,Animal Production Research Center Nitra | Harrath A.H.,King Saud University
Reproductive Toxicology | Year: 2017

The petroleum low-weight aromatic hydrocarbons benzene, toluene, ethylbenzene, m/p-xylene, and o-xylene, also known as BTEX, are among the most common hazardous sources of environmental contamination. This paper reviews the available data concerning the effects of BTEX on different aspects of female reproduction, including the fecundity, ovaries, central nervous system (CNS), oocytes, embryos, oviducts, cytogenetics of somatic and generative cells, intracellular signaling systems, and hypothalamic, pituitary and peripheral reproductive hormones. Analysis of the available literature demonstrates that BTEX can exert negative effects on various female reproductive sites, including the CNS-pituitary-ovarian axis, their signaling molecules and receptors, ovarian follicles, corpora lutea, oocytes, embryos, oviducts, ovarian cycles, fertility, and the viability of offspring. These effects could be due to the ability of BTEX to destroy chromosomes, to affect cell metabolism, including the accumulation of free radicals, and to affect the release of hormonal regulators of reproductive processes and intracellular protein kinases. © 2017 Elsevier Inc.

Spalekova E.,Animal Production Research Center Nitra | Makarevich A.V.,Animal Production Research Center Nitra | Kubovicova E.,Animal Production Research Center Nitra | Ostro A.,University of P.J. Šafarik | And 2 more authors.
Acta Veterinaria Brno | Year: 2014

Caffeine is a well-known sperm motility stimulator, however, its effects on cooling-stored ram semen are unknown. The aim of the study was to examine the effect of caffeine on selected motility and viability indices of cooling-stored ram spermatozoa. Sperm ejaculates from 4 rams were diluted (1:3) in a Triladyl extender. Samples were stored for 96 h at 4-5 °C in two sets. In the first set used for motility analysis, caffeine at concentrations of 1, 2 or 4 mmol·l-1 was added to sperm aliquots on the day of analysis. In the second set used for viability assay, caffeine at the same concentrations (1, 2 or 4 mmol·l-1) was added at the beginning of storage. Control was left without caffeine addition. Sperm motility was analyzed at 0, 24, 48 and 72 h of cooling-storage. Viability assays were done after 72-96 h of cooling-storage. Caffeine significantly (P < 0.05) increased sperm motility and progressive movement and maintained this value for 72 h. Caffeine at the dose of 2 mmol·l-1 and 4 mmol·l-1 significantly (P < 0.05) reduced the proportion of dead/necrotic sperm detected by propidium iodide and proportion of apoptotic sperm detected by Yo-Pro-1, respectively. No effect of caffeine on plasma membrane integrity was noted. Proportion of sperm with membrane destabilization (annexin V-Fluos) was reduced by caffeine given at 1 and 4 mmol·l-1 compared to control. Our study for the first time demonstrates that caffeine maintains motility and viability of cooling-stored ram sperm for longer time compared to control.

Makarevich A.V.,Animal Production Research Center Nitra | Kubovicova E.,Animal Production Research Center Nitra | Sirotkin A.V.,Animal Production Research Center Nitra | Pivko J.,Animal Production Research Center Nitra
Veterinarni Medicina | Year: 2010

The goal of this study was to examine the effect of epidermal growth factor (EGF) on sperm viability using two fluorescent techniques and to analyze the obtained results in relation to sperm motility, determined by subjective estimation. Fresh ram semen diluted in a Biladyl commercial extender was cooling stored (at 4 °C in a fridge) for four days in the presence of EGF at doses of 0, 10, 100, 200 or 400 ng/ml. Thereafter, sperm samples were analyzed for progressive motility (Motility test) and membrane integrity using two fluorescent techniques: SYBR-14/PI (Method 1) or PI/DAPI (Method 2). Application of Method 1 did not detect an effect of EGF at any concentration on sperm membrane integrity. A positive effect of EGF (200 ng/ml) on sperm membrane integrity was found using Method 2 of staining, and this result was confirmed by the sperm motility test, which demonstrated an EGF-stimulating effect (200 or 400 ng/ml) on a percentage of progressively moving spermatozoa. Strong positive correlations between Methods 1 and 2 (r = 0.785), Method 1 and Motility (r = 0.803), Method 2 and Motility (r = 0.699), as well as between both techniques taken together and the Motility test (r = 0.853) were found. Regression analysis confirmed that Method 2 was more exact than Method 1, and the results obtained with Method 2 are comparable with those of the Motility test. Dependence of the viability or motility on EGF concentrations (linear regression function) was significant only for Method 2 or the Motility test. The obtained results suggest a stimulating effect of EGF (at higher concentrations) on ram sperm functions (viability/membrane integrity and motility). Furthermore, they indicate substantial differences between two fluorescent techniques in the determination of sperm membrane integrity. Only the data obtained using PI/DAPI were confirmed by a functional Motility test. These findings suggest that the technique chosen for analysis of sperm viability can influence the conclusion concerning the effects of the treatment on sperm function.

Roychoudhury S.,Assam University | Bulla J.,Slovak University of Agriculture | Sirotkin A.V.,Animal Production Research Center Nitra | Kolesarova A.,Slovak University of Agriculture
Journal of Environmental Science and Health - Part A Toxic/Hazardous Substances and Environmental Engineering | Year: 2014

Objective of this in vitro study was to examine the secretion activity (progesterone and insulin-like growth factor I) of porcine ovarian granulosa cells after copper (Cu) addition and to outline a potential intracellular mediator (cyclin B1) of its effects. It also aimed at investigating the apoptotic potential of Cu on porcine ovarian granulosa cells after addition in vitro. Ovarian granulosa cells were incubated with copper sulphate (CuSO 4·5H2O) at the doses 0.33, 0.40, 0.50, 1.0 and 2.0 μL mL-1 for 18 h and compared with control group without Cu addition. Release of progesterone (P4) and insulin-like growth factor I (IGF-I) by granulosa cells was assessed by RIA, expression of cyclin B1 by immunocytochemistry and apoptosis by TUNEL assay. Observations show that P 4 release by granulosa cells was inhibited while the release of IGF-I and cyclin B1 was stimulated significantly (P < 0.05) by CuSO 4·5H2O addition at the dose 2.0 μL mL -1. Also, addition of CuSO4.5H2O at the lowest dose used in the study (0.33 μL mL-1) significantly (P < 0.05) decreased apoptosis in granulosa cells. In conclusion, results indicate dose dependent effect of Cu on (1) secretion of steroid hormone progesterone and growth factor IGF-I, (2) expression of cyclin B1 as marker of proliferation of porcine ovarian granulosa cells, (3) apoptosis of porcine ovarian granulosa cells and, (4) that the effect of Cu on ovarian cell proliferation could be mediated by IGF-I and cyclin B1. Obtained data suggest interference of Cu in the pathways of proliferation of porcine ovarian granulosa cells through hormonal and intracellular peptide cyclin B1. © 2014 Copyright Taylor and Francis Group, LLC.

Bahelka I.,Animal Production Research Center Nitra | Gondekova M.,Animal Production Research Center Nitra
Journal of Central European Agriculture | Year: 2016

The present study aimed at comparison of chemical composition, meat quality and sensory parameters of cow’s meat and meat from bulls produced under Slovak conditions and sold in retail of Slovakia. The analysis was performed on 181 cows and 78 bulls. Cows were also divided in two groups according to age at slaughter-over (n = 135) and/or under 4 years (n = 46). The meat samples were taken in eight slaughter houses located in the western, central and eastern part of Slovakia. The age of cows slaughtered had significant effect (P < 0.05) on the most meat quality and sensory traits which were evaluated much more worse in older cows with comparison to bulls than in younger cows. Older cows had lower content of proteins and water in meat, lower pH7 and colour parameter “L” as well as worse evaluation of all sensory parameters in comparison to bulls (P < 0.05). On the other hand, intramuscular fat content, energetic value of meat, marbling, pH48, colour parameter “a” and cooking loss were higher in meat of older cows than bulls (P < 0.05). Differences in traits observed between younger cows and bulls were statistically significant (P < 0.05) only for content of proteins and water, pH48, colour parameter “a”, cooking loss and evaluation of odour. The hypothesis of significantly poorer meat and eating quality is justified in the case of cows over 4 years. The study did not confirm the decreasing age of cows at slaughter as suggested previous studies. © 2016, University of Zagreb - Faculty of Agriculture. All rights reserved.

Sirotkin A.V.,Constantine the Philosopher University | Sirotkin A.V.,Animal Production Research Center Nitra | Dekanova P.,Constantine the Philosopher University | Harrath A.H.,King Saud University | And 2 more authors.
Cell and Tissue Research | Year: 2014

The roles of the mTOR system enzyme sirtuin 1 (SIRT1), the transcription factor p53 and the nuclear factor kappaB (NF-κB) and their interrelationships in the control of ovarian function have not been well studied. We examine, in vitro, the involvement of SIRT1, p53 and the p65 and p50 subunits of NFκB and their interrelationships in the control of the apoptosis and proliferation of porcine ovarian granulosa cells. Monolayers of primary granulosa cells were transfected with cDNA constructs encoding SIRT1, p53, p65 or p50 alone or were co-transfected with gene constructs for SIRT1 together with p53, p65 or p50. The accumulation of SIRT1, markers of proliferation (mitogen-activated protein kinase or extracellular-signal-regulated kinases 1,2) and a marker of apoptosis (caspase 3) was detected by immunocytochemistry. Transfection of cells with a SIRT1 gene construct alone promoted the accumulation of SIRT1 and decreased the accumulation of proliferation markers but did not affect the marker of apoptosis. Transfection of cells with gene constructs encoding p53, p50 or p65 decreased the expression of proliferation markers but not the apoptosis marker. Co-transfection of cells with SIRT1 cDNA changed the action of p65 on cell proliferation from inhibitory to stimulatory. SIRT1 overexpression induced the pro-apoptotic action of p53 and p50 but not of p65 constructs. Thus, SIRT1, p53 and NF-κB are involved in the control of both the proliferation and the apoptosis of ovarian cells. These novel data on the cross-talk between the mTOR/SIRT1 system and the transcription factors p53 and NF-κB show both the inhibitory (proliferation) and stimulatory (apoptosis) influences of SIRT1 on transcription factor action in ovarian cells. © 2014, Springer-Verlag Berlin Heidelberg.

Sirotkin A.V.,Animal Production Research Center Nitra | Sirotkin A.V.,Constantine the Philosopher University | Benco A.,Constantine the Philosopher University | Tandlmajerova A.,Constantine the Philosopher University | Vasicek D.,Animal Production Research Center Nitra
Cell Proliferation | Year: 2012

The aim of our in vitro experiments was to examine the role of transcription factor p53 and the metabolic hormone leptin, in controlling basic functions (proliferation, apoptosis and secretory activity) of ovarian cells, as well as involvement of p53 in mediating or modulating actions of leptin, on ovarian cells. Porcine ovarian granulosa cells, transfected and non-transfected with a gene construct encoding p53, were cultured with leptin (at concentrations of 0, 1, 10 or 100ng/ml). Accumulation of p53 and of apoptosis-related (bax) and proliferation-related (PCNA, cyclin B1) substances was evaluated by SDS-PAGE-western blotting. Secretion of progesterone (P4) was measured by RIA. Transfection with the p53 gene construct promoted accumulation of this transcription factor within cells. It also stimulated expression of bax (which can be thought of as a marker of apoptosis), and reduced accumulation of proliferation-related substances PCNA and cyclin B1. Overexpression of p53 resulted in reduced P4 secretion. Leptin, when added alone, increased accumulation of p53, bax and PCNA, decreased accumulation of cyclin B1 and had no effect on P4 secretion. Transfection of cells with p53 gene construct reversed effects of leptin on cyclin B1 and induced stimulatory effects of leptin on P4 release, but did not modify leptin action on p53, bax and PCNA. These multiple effects of the p53 gene construct on granulosa cells, cultured with and without leptin, (i) demonstrate that leptin can be involved in control of porcine ovarian cell proliferation, apoptosis and expression of p53, but not on P4 release; and (ii) confirm involvement of p53 in promoting apoptosis and suppression of proliferation and P4 secretion in these cells. (iii) The similarity of p53 and leptin's actions on bax and cyclin B1, and inability of p53 to further promote leptin action on this parameter suggest that p53 can be a mediator of leptin's action on ovarian cell apoptosis. (iv) On the other hand, p53 can modulate, but probably not mediate the effects of leptin on ovarian cell proliferation and P4 release. © 2011 Blackwell Publishing Ltd.

Sirotkin A.V.,Animal Production Research Center Nitra | Sirotkin A.V.,Constantine the Philosopher University | Sirotkin A.V.,Institute of Animal Genetics and Reproduction
International Journal of Biochemistry and Cell Biology | Year: 2011

The present short review demonstrates an important role of different cytokines (colony stimulating factors, tumor necrosis factors, interleukins, anti-Mullerian hormone, inhibin, activin, follistatin, bone morphogenetic proteins, growth and differentiation factors) in the control of different ovarian functions - ovarian cell proliferation, apoptosis, folliculogenesis, luteogenesis, oogenesis, release of hormones, response to upstream hormonal regulators, fertility and, in some cases, in development of ovarian disorders. The possibility of practical application of these molecules for characterization, prediction and regulation of the ovarian state including treatment of ovarian disorders is demonstrated. © 2011 Elsevier Ltd.

Cikos S.,Slovak Academy of Sciences | Fabian D.,Slovak Academy of Sciences | Makarevich A.V.,Animal Production Research Center Nitra | Chrenek P.,Animal Production Research Center Nitra | And 2 more authors.
Human Reproduction | Year: 2011

Background The involvement of biogenic monoamines in early ('preneural') embryogenesis has been well documented in lower vertebrates, but much less information is available about the role of these molecules in the earliest stages of development in mammals, including humans. Methods Databases (PubMed, ISI Web of Knowledge and Scopus) were searched for studies relating to biogenic monoamines functioning in early embryos. The available data on the expression of histamine, serotonin and adrenergic receptors during mammalian preimplantation development were summarized, and the potential roles of biogenic monoamines in very early pregnancy were discussed. Results The roles of biogenic monoamines in mammalian preimplantation embryo development can be diverse, depending on the embryo developmental stage, and the physiological status of the maternal organism. Several receptors for biogenic monoamines are expressed and biologically functional in cells of preimplantation embryos. Activation of histamine receptors can play a role in embryo implantation and trophoblast invasion. Activation of adrenergic and serotonin receptors can influence proliferation and survival of early embryonic cells. Conclusions Biogenic monoamines can play an important role in physiological conditions, contributing to embryo-maternal interactions, or can influence the early embryo under unfavorable or pathological conditions (e.g. in maternal stress, or in women taking certain antidepressants, anti-migraine or anti-ulcer drugs). © 2011 The Author.

Sirotkin A.V.,Animal Production Research Center Nitra | Sirotkin A.V.,Constantine the Philosopher University | Bauer M.,Animal Production Research Center Nitra | Bauer M.,Constantine the Philosopher University
Cell Stress and Chaperones | Year: 2011

The present studies aimed to understand the interrelationships between stress, hormones and heat shock proteins (HSPs) in the ovary. We examined (1) whether HSP70.2, HSP72 and HSP105/110 can be produced and accumulated in porcine ovarian tissue, (2) whether these HSPs could be indicators of stress, i.e. whether two kinds of stress (high temperatures and malnutrition/serum deprivation) can affect them, and (3) whether some hormonal regulators of ovarian functions (insulin-like growth factor (IGF)-I, leptin and follicle-stimulating hormone (FSH)) can affect these HSPs and response of ovaries to HSP-related stress. We analysed the expression of HSP70.2, HSP72 and HSP105/110 mRNA (by using real-time reverse transcriptase polymerase chain reaction) in porcine ovarian granulosa cells, as well as the accumulation of HSP70 protein (by using sodium dodecyl sulphate polyacrylamide gel electrophoresis-Western) in either whole ovarian follicles and granulose cells cultured at normal (37.5°C) or high (41.5°C) temperature, with and without serum and with and without IGF-I, leptin and FSH. Expression of mRNA for HSP70.2, HSP72 and HSP105/110 in ovarian granulosa cells and accumulation of HSP70 protein in whole ovarian follicles and granulosa cells were demonstrated. In all the groups, addition of either IGF-I, leptin and FSH reduced the expression of HSP70.2, HSP72 and HSP105/110 mRNA. Both high temperature, serum deprivation and their combination resulted in increase in mRNAs for all three analysed HSPs. Additions of either IGF-I, leptin and FSH prevented the stimulatory effect of both high temperature and serum deprivation on the transcription of HSP70.2, HSP72 and HSP105/110. In contrast, high temperature reduced accumulation of peptide HSP70 in both ovarian follicles and granulosa cell. Serum deprivation promoted accumulation of HSP70 in granulosa cells, but not in ovarian follicles. Addition of IGF-I, leptin and FSH was able to alter accumulation of HSP70 in both follicles and granulosa cells. The present observations suggest (1) that HSPs can be synthesised in ovarian follicular granulosa cells; (2) that hormones (IGF-I, leptin and FSH) can inhibit, whilst stressors (both high temperature and malnutrition/serum deprivation) can stimulate transcription of HSP70.2, HSP72 and HSP105/ 110 genes, whilst heat stress, but not malnutrition, can promote depletion of HSP70 in ovarian cells, and (3) that hormones (IGF-I, leptin and FSH) can prevent stress-related changes in HSPs. The application of HSPs as indicators and mediators of stress and hormones on ovarian functions, as well as use of hormones and HSPs as anti-stressor molecules, are discussed.. © 2010 Cell Stress Society International.

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