Tetrabromobisphenol A (TBBPA) degradation was investigated using white rot fungi and their oxidative enzymes. Strains of the Trametes, Pleurotus, Bjerkandera and Dichomitus genera eliminated almost 1 mM TBBPA within 4 days. Laccase, whose role in TBBPA degradation was demonstrated in fungal cultures, was applied to TBBPA degradation alone and in combination with cellobiose dehydrogenase from Sclerotium rolfsii. Purified laccase from Trametes versicolor degraded approximately 2 mM TBBPA within 5 h, while the addition of cellobiose dehydrogenase increased the degradation rate to almost 2.5 mM within 3 h. Laccase was used to prepare TBBPA metabolites 2,6-dibromo-4-(2-hydroxypropane-2-yl) phenol (1), 2,6-dibromo-4-(2-methoxypropane-2-yl) phenol (2) and 1-(3,5-dibromo-4-hydroxyphen-1-yl)-2,2',6,6'-tetrabromo-4,4'-isopropylidene diphenol (3). As compounds 1 and 3 were identical to the TBBPA metabolites prepared by using rat and human liver fractions (Zalko et al., 2006), laccase can provide a simple means of preparing these metabolites for toxicity studies. Products 1 and 2 exhibited estrogenic effects, unlike TBBPA, but lower cell toxicity.
Tetrabromobisphenol A (TBBPA) is the main flame retardant used in printed circuit boards and laminates. The human population is highly exposed to TBBPA as it is used in consumer electronics as well as office and communication equipment. The main use of hexabromocyclododecane (HBCD) is in insulation foam boards, which are widely used in the construction sector. Brominated flame retardants may possess endocrine disrupting activity and thus represent a threat to the environment, including humans and their reproduction. The aim of this work was to evaluate the oestrogenic effects of TBBPA and HBCD in vitro on MCF-7 cells. We used the proliferation test (E-screen assay) in MCF-7 breast cancer cells and reverse transcription quantitative polymerase chain reaction analysis of TFF1 gene expression to analyse oestrogenicity of the studied compounds. RT-qPCR has proved to be a fast and valuable molecular technique in gene expression quantification. HBCD but not TBBPA increased cell proliferation in MCF-7 cells and up-regulated TFF1 gene expression in a concentration-dependent manner. Anti-oestrogen ICI 182,780 inhibited up-regulation of TFF1 by HBCD. We have shown that HBCD displays oestrogen- like effects on MCF-7 cells. TBBPA, on the other hand, has not shown any oestrogenic effect mediated by the oestrogen receptor ?.
- MeSH
- alfa receptor estrogenů metabolismus MeSH
- antagonisté estrogenu toxicita MeSH
- bromované uhlovodíky toxicita MeSH
- endokrinní disruptory toxicita MeSH
- estriol analogy a deriváty metabolismus MeSH
- estrogeny analýza MeSH
- exprese genu MeSH
- financování organizované MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nádorové supresorové proteiny metabolismus MeSH
- polybrombifenylové sloučeniny toxicita MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- proliferace buněk MeSH
- retardanty hoření toxicita MeSH
- upregulace MeSH
- Check Tag
- lidé MeSH
The effect of a low dose of vinclozolin within the development of the reproductive tract during gestation (VIN-GD 15-22) and puberty (VIN-PND 23-44) in CD1 mice was tested. We found a decrease in the anogenital distance, prostate weight and pathology of testes in both experimental groups. Sperm counts decreased to 46% (VIN-GD) and to 81% (VIN-PND), and also the acrosomal state (evaluated by antiacrosomal antibody) decreased in both groups to 89% in comparison to the control group (100%). Sperm head abnormalities increased by approximately 18% and 13%, respectively. In this connection, the expression of some genes was changed (arosome-related gene (Acr), apoptosis related genes (p53, p21)). In conclusion, a low dose of vinclozolin affected the reproductive tract, sperm parameters and expression of selected genes in both experimental groups.
- MeSH
- antagonisté androgenů toxicita MeSH
- fungicidy průmyslové toxicita MeSH
- mužské pohlavní orgány embryologie účinky léků MeSH
- myši inbrední ICR MeSH
- myši MeSH
- oxazoly toxicita MeSH
- plod účinky léků MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- spermatogeneze MeSH
- těhotenství MeSH
- tělesná hmotnost účinky léků MeSH
- testis metabolismus účinky léků MeSH
- ubikvitin metabolismus MeSH
- velikost orgánu účinky léků MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
The effect of low dose of fungicide vinclozolin within development of reproductive tract during gestation (GD) and puberty (PND) in outbred CD1 mice was tested. We found a decrease in the anogenital distance, prostate weight and pathology of testes in both experimental groups. Marked negative influence has had on sperm parameters. Sperm counts decreased to 46% (GD) and to 81% (PND) and also acrosomal state (evaluated by antiacrosomal antibody) decreased in both groups to 89% in comparison to control group (100%). Sperm head abnormalities increased about 18% and 13%, respectively. In this connection the expression some genes was changed (arosome– related gene (Acr), apoptosis related genes (p53, p21)). In conclusion, low dose of vinclozolin affected the reproductive tract, sperm parameters and expression of selected genes in both experimental groups. The work was supported by the Grants of the Ministry of Education of the Czech Republic, grant No. 1M06011 and 2B06151 and in part by the Institutional Research Support AVOZ 50520701.
BACKGROUND: Antibodies to human sperm are useful diagnostic reagents for detection of changes in sperm protein expression and their relationship with sperm defects and male infertility. The specificity of Hs-16 monoclonal antibody (mAb) and the localization and frequency of the occurrence of Hs-16-recognized protein on human spermatozoa were investigated. METHODS: Samples from 30 fertile men with normal spermiograms and 30 men with pathological spermiograms were studied. The specificity of Hs-16 mAb was analysed by the western blotting technique and matrix-assisted laser desorption/ionization mass spectrometry. Indirect immunofluorescence with Hs-16 antibody was used to test sperm ejaculates. RESULTS: The Hs-16 antibody detected a human sperm and seminal plasma protein, which was determined to be secretory actin-binding protein (SABP). This specificity was also verified by co-localization of SABP and actin on spermatozoa with Hs-16 and anti-actin antibodies, and partial co-localization of these proteins was found. SABP was localized on the sperm tail, mainly in the midpiece of the tail. Other parts of spermatozoa were labelled with lower frequency. A significant difference was found in SABP labelling between men with normal spermiograms and donors with asthenozoospermia or oligoasthenoteratozoospermia (both P < 0.01), and asthenozoospermia versus oligoasthenoteratozoospermia (P < 0.05). Increased expression of SABP was observed in men with pathological spermiograms. CONCLUSIONS: Hs-16 antibody reacts specifically with SABP. SABP can serve as a marker of defective sperm and may be associated with fertility failure.
- MeSH
- aktiny analýza MeSH
- dospělí MeSH
- financování organizované MeSH
- fluorescenční protilátková technika nepřímá MeSH
- lidé MeSH
- mikrofilamentové proteiny biosyntéza sekrece MeSH
- monoklonální protilátky MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- spermie imunologie metabolismus MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
Seminal plasma proteins bind the sperm surface at ejaculation and may modulate several aspects of sperm activity during reproduction. DQH sperm surface protein, present in boar seminal plasma, shows affinity to phoshorylcholine, acidic polysaccharides, oviductal epithelium and zona pellucida glycoproteins. Monoclonal antibodies (MAbs) against DQH protein were prepared and used for determination of the DQH protein origin in boar reproductive organs, its localization on boar spermatozoa, and for investigation of its binding abilities in the porcine oviduct and to the zona pellucida of the oocyte. The mRNA transcript of DQH protein was found in seminal vesicles and not in the testis, epididymis and prostate. Its translated products were immunodetected by MAbs in seminal vesicle extract and fluid, in seminal vesicle tissue sections and on the membrane-associated acrosomal part of ejaculated spermatozoa. These results confirm the ability of DQH protein to bind the sperm surface at ejaculation and to participate in formation of the sperm reservoir in the porcine oviduct. Moreover, monoclonal antibodies reduced binding of sperm to oocytes and proved the role of DQH protein in the sperm-zona pellucida primary binding.
- MeSH
- fertilizace MeSH
- financování organizované MeSH
- imunohistochemie MeSH
- interakce spermie a vajíčka MeSH
- lidé MeSH
- membránové glykoproteiny genetika imunologie metabolismus MeSH
- monoklonální protilátky MeSH
- mužské pohlavní orgány metabolismus MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- prasata MeSH
- proteiny semenné plazmy metabolismus MeSH
- sperma MeSH
- spermie metabolismus MeSH
- vazba proteinů MeSH
- vejcovody metabolismus MeSH
- zona pellucida metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
The enzyme creatine kinase (CK) plays a key role in energy homeostasis of cells with high and fluctuating energy requirements. As for spermatozoa, the activity of phosphocreatine shuttle, which directs energy from the mitochondria to sites of ATP consumption, is dependent on individual species. High activities of CK are observed in spermatozoa of nonvertebrate, fish, and birds, contrary to the low-level CK activity in mammalian spermatozoa. A new monoclonal antibody (MAb) to carp sperm creatine kinase was prepared. This antibody is applicable to large-scale immunochemical techniques. In this study, it was applied to the study of carp sperm motility, and the evaluation of the influence of CK on the quality and fertilization ability of carp spermatozoa.
OBJECTIVE: To determine whether varied human spermatozoa, as detected with monoclonal antibodies against acrosomal proteins, have an influence on fertilization, transfer, pregnancy, and implantation rates when intracytoplasmic sperm injection is used. DESIGN: A retrospective study. SETTING: A private IVF center and academic research laboratory. PATIENT(S): One thousand two hundred forty men participating in the intracytoplasmic sperm injection program. INTERVENTION(S): Sperm were divided into seven groups: oligozoospermia, oligoasthenozoospermia, and oligoasthenoteratozoospermia and fresh and frozen-thawed epididymal and fresh and frozen-thawed testicular sperm. Fertilization, transfer, pregnancy, and implantation rates were recorded in each category. Sperm were tested with antibodies for detection of the of the sperm acrosome. MAIN OUTCOME MEASURE(S): Fertilization, transfer, pregnancy and implantation rates, and percentage of acrosome-reacted cells. RESULT(S): The fertilization rate and statistical evaluation showed differences between morphologically normal and pathological sperm and other groups. The freezing-thawing procedure had no influence on the fertilization of testicular sperm, but epididymal frozen-thawed sperm had a higher fertilization rate. Immunofluorescence proved decreasing sperm quality in all groups compared with the control group. This difference is not manifested in other parameters (transfer, pregnancy, implantation rates). CONCLUSION(S): The spermatozoa with varied semen characteristics and good quality, also detected with specific antibodies, gave the best fertilization rates. The paternal effect is not proved in other parameters.
- MeSH
- akrozom imunologie MeSH
- financování organizované MeSH
- hodnocení výsledků zdravotní péče metody MeSH
- imunoanalýza metody MeSH
- imunokomplex analýza MeSH
- intracytoplazmatické injekce spermie statistika a číselné údaje MeSH
- kultivované buňky MeSH
- lidé MeSH
- mužská infertilita epidemiologie imunologie terapie MeSH
- retrospektivní studie MeSH
- sperma cytologie imunologie MeSH
- těhotenství MeSH
- úhrn těhotenství na počet žen v reprodukčním věku MeSH
- výsledek terapie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
