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Males-females differences in the spectrum of chromosomal aberrations in the group of nanocomposites production workers
Rössnerová, Andrea ; Pelcová, D. ; Ždímal, Vladimír ; Elzeinova, Fatima ; Margaryan, Hasmik ; Chvojková, Irena ; Topinka, Jan ; Schwarz, Jaroslav ; Ondráček, Jakub ; Koštejn, Martin ; Komarc, M. ; Vlčková, Š. ; Fenclová, Z. ; Lischková, L. ; Dvořáčková, Š. ; Rössner ml., Pavel
An increase in the use of nanomaterials (NM) has been witnessed in many areas of human life. Therefore, assessment of genotoxicity of NM and nanoparticles (NP) is one of the main objectives of genetic toxicology. Despite this fact, human cytogenetic studies following the exposure to NP are still rare. Moreover, no relevant information on possible differences in sensitivity to NP related to gender is available.\n\nIn this study we periodically (in September 2016, 2017 and 2018; pre-shift and post-shift each year) analyzed a group of workers (both genders), working long time in nanocomposites research, and matched controls. Aerosol exposure monitoring of particulate matter including nano-sized fractions was carried out during working shift. Micronucleus assay using Human Pan Centromeric probes, was applied to distinguish, besides the frequency of total MN in binucleated cells (BNC), also other types of chromosomal damage (losses and breaks). Moreover, whole-chromosome painting (WCP) for autosome #1 and both gonosomes (X and Y) were applied in third sampling period (2018) with the aim to identify the particular structural and numerical chromosomal aberrations.\n\nObtained results showed: (i) differences in the risk of exposure to NP related to individual working processes (welding, smelting and machining); (ii) differences in chemical composition of nano-fraction; (iii) no effect of chronic exposure of NP (total MN) opposite to significant effect of acute exposure; (iv) gender-related DNA damage differences (females seem to be more sensitive to chromosomal losses). Additional data from WCP suggested increased frequency of numerical aberrations in gonosomes.
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DNA damage induced by occupational exposure to copper oxide nanoparticles
Rössner st., Pavel ; Pelcová, D. ; Elzeinová, Fatima ; Mikuška, Pavel ; Večeřa, Zbyněk ; Coufalík, Pavel ; Vlčková, Š. ; Fenclová, Z. ; Rössnerová, Andrea
Copper oxide nanoparticles (CuO NPs) have a widespread use in industry, chemistry, in production of electronic devices and as an antimicrobial agent. Although copper is an important biogenic element, CuO NPs are toxic with the ability to induce oxidative stress, apoptosis, cell cycle arrest or DNA damage. For humans, the inhalation route is the most common way of exposure to CuO NPs. In the body, CuO NPs may be either deposited in the lungs, or transported to other organs. Their presence usually causes oxidative stress or inflammatory responses, consequently leading to DNA damage. In this study, we investigated the effect of CuO NPs inhalation on DNA damage in a group of researches conducting animal exposure experiments. The subjects were exposed to various metal oxide nanoparticles, including CuO NPs, by inhalation for an average of 4.9 ± 0.4 years. The average mass concentration of Cu in the air during the experiment was 7.3 ± 3.2 ng/m3. Subjects not exposed to nanoparticles served as a control group. We applied micronucleus assay using Human Pan Centromeric probes to detect DNA damage and to distinguish between the frequency of centromere positive (CEN+) and centromere negative (CEN−) micronuclei (MN) in the binucleated cells. We\ndid not find differences between both groups for either mean MN frequency (10.38 ± 2.50 vs. 11.88 ± 3.01 MN/1000 binucleated cells), or CEN+/CEN- ratio (58%/42% vs. 55%/45%), for the exposed and controls, respectively. In conclusion, inhalation of CuO NPs at this low-level exposure had no effect on chromosomal losses and/or breaks.
Diabetes mellitus negatively affects male reproductive parameters in vivo
Valášková, Eliška ; Žatecká, Eva ; Pavlínková, Gabriela ; Bohuslavová, Romana ; Dorosh, Andriy ; Elzeinová, Fatima ; Kubátová, Alena ; Margaryan, Hasmik ; Pěknicová, Jana
According to the World Health Organization (WHO), 15% of couples in reproductive age suffer from infertility problems, and up to 60% of cases are caused by male factor. This could be caused by genetic background, environmental factors and various diseases, including diabetes mellitus (DM). However, the impact of DM on male fertility is not fully understood. . The aim of this study was to investigate the effects of DM on reproductive parameters and sperm quality, using mouse model. DM (type 1) was induced by Streptozotocin in FVB inbred mouse strain. Mice with blood sugar levels higher than 13.9 mmol/L were considered diabetic. After 4 weeks of diabetes exposure, diabetic males were bred with wild type females and transgenerational effect of DM was assessed. Selected morphological, cellular, and molecular parameters of diabetic males and their male offspring were compared to appropriate controls. There was an increased in sperm fragmentation and abnormalities of sperm morphology in diabetic mice in both generations. An increased staining with apoptotic marker annexin V was also detected in the diabetic groups. Furthermore, a presence of protamines as major sperm nuclear proteins was analysed. Protamine 1 to protamine 2 ratio (P1/P2), a marker of male fertility, was altered in sperms of experimental diabetic animals in both generations. Our findings indicate that DM type 1 negatively affects sperm quality and P1/P2 ratio and this negative effect is transmitted to the progeny
Book of abstract of XXIInd Symposium of immunology and biology of reproduction
Pěknicová, Jana ; Elzeinová, Fatima ; Kubátová, Alena
Book of abstracts. The Symposium was focused on immunology of reproduction and specific problems in reproduction (mainly in human infertility).
Book of abstract XXIst Symposium of biology and immunology of reproduction
Pěknicová, Jana ; Kubátová, Alena ; Elzeinová, Fatima
The Symposium was focused on immunology of reproduction and specific problems in reproduction (mainly in human infertility).
Book of abstracts XXth Symposium of Biology and Immunology of Reproduction with international participation
Pěknicová, Jana ; Kubátová, Alena ; Elzeinová, Fatima
Book of abstracts. The Symposium was focused on immunology of reproduction and specific problems in reproduction (mainly in human infertility).
Effect of diabetes mellitus on reproductive parameters in mice
Margaryan, Hasmik ; Elzeinová, Fatima ; Kubátová, Alena ; Strolená, Eva ; Pěknicová, Jana
The decrease in population fertility has become a major concern in many developed countries. Recent studies show that infertility is affecting an estimated 15% of all couples (World Health Organization, WHO, 2010). Male infertility is the primary or contributing cause in 60% of cases. Male infertility is caused by a number of factors, such as genetic background, various environmental factors and disease. Diabetes mellitus (DM), a serious health problem on its own, is also suspected to be a contributing factor to male infertility. The aim of this project was to analyze the cellular, molecular and genetic effects of diabetic environment on spermatogenesis and sperm quality and to determine the impact of DM on the in vivo reproduction, using the mouse model (Mus musculus) inbred FVB. Diabetes was induced using streptozotocin. We used our knowledge and tools (unique monoclonal antibodies developed by our group) to determine the status of reproductive organs, anogenital distance, and the quality of sperms. Genetic analysis was performed by a quantitative Reverse Transcription Polymerase Chain Reaction (qPCR). We tested selected genes which are expressed in testicular tissue and thus can influence process of spermatogenesis and consequently the sperm quality. Our preliminary data strongly suggest that DM impairs male fertility. We have found significant changes in the body and reproductive organ weight of mice with DM. We have identified qualitative and quantitative changes in the expression of proteins in epididymal fluid and sperms. We have also detected an increased number of apoptotic cells in sperm of diabetic mice compared to the control group. To our knowledge, there is no study assessing the correlation between DM and “unexplained infertility”. In view of this, it is essential to analyze the effects of DM on male fertility, sperm quality, and reproduction parameters.
Expression of estrogen receptor beta (ERβ) In murine male reproductive tract and sperm
Dostálová, Pavla ; Děd, Lukáš ; Dorosh, Andriy ; Elzeinová, Fatima ; Pěknicová, Jana
Estrogens are steroid hormones that play an important role in reproduction of both sexes. In male, the main source of estrogens are testes where both somatic and germ cells are responsible for testosteron conversion to estrogens. Estrogens are involved in control of spermatogenesis, fluid reabsorption in rete testis and epididymis, and in later maturation steps that sperm undergo in female genital tract (capacitation, acrosome reaction). Generally, estrogen action is mediated through binding to estrogen receptors (ERs) which than lead to classical genomic or rapid non-genomic signaling. Nowadays, two classical estrogen receptors are known – ERα and ERβ. ERβ is a predominant variant in testes, while ERα is more abundant in rete testis and initial segment of epididymis. In addition to classical ERs, several splice variants that can differ in their ligand- or DNA-binding properties were detected in different tissues and cell lines. ERs mostly work as a dimer (homo- and hetero-) and splice variants often „only“ modulate function of classical full-length ERs. Therefore, estrogen action seems to be a very complex. To contribute to understanding of estrogen action in male, we detected ERβ and its potential splice variants in mice testis, epididymis and sperm. According to our results, two variants are present in all analysed tissues and cells. These variants differ in one exon in ligand binding domain which leads to different affinity for estrogens. To analyse these variants also at a protein level, we prepared specific monoclonal antibodies recognizing particular variant of ERβ. Both atibodies detected band(s) in protein extracts from testes or epididymis. Taking together, there are at least two variants of ERβ in mice testes, epididymis and sperm and it seems that both variants are similar in abundance within the same organ or sperm.
Effect of exposure to bisphenol A on gene expression in the testicular tissue in male mice
Dorosh, Andriy ; Elzeinová, Fatima ; Žatecká, Eva ; Kubátová, Alena ; Pěknicová, Jana
Laboratory of Reproductive Biology, Institute of Biotechnology, Academy of Sciences of the Czech Republic, v.v.i., Prague, Czech Republic Bisphenol A (BPA) is a synthetic, endocrine-disrupting compound able to directly bind estrogen receptors. Free BPA has been detected in human samples indicating that humans are internally exposed to BPA. The purpose of this study was to analyse the effect of BPA on the male reproductive system and testicular gene expression in germ cells. We studied the influence of long-term low concentration BPA exposition on male fertility in vivo in a two-generation study in C57BL/6 mouse strain. In this work, BPA was added with water at two environmentally relevant concentrations: 0, 4 and 4 µg/l. We measured the reproductive organs weight and sperm cells morphology and quality. Expression of genes involved in endocrine regulation and energy metabolism in testis was analysed after BPA exposure. Next, the epigenetic mechanisms of gene expression and regulation during the germ cell differentiation and effect of BPA will be analysed.
The effect of tetrabromobisphenol a on protamination and DNA quality of mouse sperm
Žatecká, Eva ; Castillo, J. ; Elzeinová, Fatima ; Kubátová, Alena ; Děd, Lukáš ; Pěknicová, Jana ; Oliva, R.
Tetrabromobisphenol (TBBPA) is a widely used brominated flame retardant, currently its consumption is 210,000 tons / year and is still growing. In our previous multigenerational in vivo study we have demonstrated that TBBPA is able to induce apoptosis of testicular cells and changes in the expression of genes important for proper spermatogenesis. However the potential effect of TBBPA on epidydimal spermatozoa had not yet been investigated. Therefore, we performed further study to evaluate the effect of on sperm DNA integrity and on the protamines as the major nuclear proteins. C57Bl/6J mice pups (n=10) were exposed to TBBPA (experimental group) during the gestation, lactation, pre-pubertal and pubertal periods up to the age of 70 days and compared control mice pups (n= 10) which were not exposed. Our results demonstrate that TBBPA treatment results in a significantly decreased P1/P2 ratio, increased total protamine/DNA ratio and increased DNA fragmentation observed between TBBPA and control mice, respectively. Protamines have recently been connected to the epigenetic marking of sperm chromatin in human and mouse spermatozoa. Thus, our findings suggest that TBBPA exposure, in addition to result in increased sperm DNA damage, may also alter the epigenetic marking of sperm chromatin.

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