Východisko. Je známo, že různé expozice mužů chemickými látkami mohou významně zhoršit kvalitu i kvantitu produkovaných spermií. Cílem naší studie bylo ověřit, zda znečištěné ovzduší v okrese Teplice má negativní vliv na kvalitu spermií u mužů žijících v tomto okrese. Metody a výsledky. Bylo vyšetřeno 325 18letých mužů žijících v okrese Teplice a v kontrolní oblasti Prachatice. Odběry vzorků proběhly v roce 1992 a 1994 vždy na konci zimy a na podzim. Podle SZO laboratorního manuálu pro vyšetření lidského semene byly stanoveny základní spermatologické parametry: objem semene, pH, pohyb, počet a morfologie spermií. U vybraných skupin mužů byla také vyšetřena frekvence aneuploidií ve spermiích. Vyšetření aneuploidií bylo provedeno pomocí tříbarevné fluorescenční in situ hybridizace s využitím satelitních DNA sond specifických pro chromozómy X, Y a 8. Pro analýzu dat byla použita logistická regrese. Byly stanoveny odd’s ratio (OR’s). Vzrůst OR’s byl zjištěn pro morfologii spermií (4,1 a 10,1 pro expozici střední, resp. vysokou), pro morfologii hlavičky (6,1 a 4,1) a u procenta motilních spermií (9,8 a 3,5). Více exponovaní muži měli zvýšenou frekvenci disomií chromozómů X (p=0,012), XY (p=0,01) a Y (p<0,001). Závěry. Použité bioindikátory toxického a genetického poškození spermií ukazují na zhoršenou kvalitu spermií u teplických mužů.

Backgrounds. It has been described that an exposition of males to chemical substances may significantly impoverish quality and quantity of produced spermatozoa. The aim of our study was to test whether the polluted air in the Teplice district has negative effects on the quality of sperm of males living in this district. Methods and Results. 325 males 18-year-old living in the Teplice district and in the control district of Prachatice were tested. Samples were taken in 1992 and 1994, always at the end of winter and in autumn. According to WHO laboratory manual for investigation of the human sperm, basic parameters were determined: volume of the semen, pH, motility, number and morphology of spermatozoa. In selected groups of males the frequency of aneuploidia of spermatozoa was also examined. Examination of aneuploidia was done using three color fluorescence in situ hybridisation with satellite DNA proves specific for X, Z and 8 chromosomes. Logistic regression was used for the data analysis and Odd’s Ratio was estimated (OR’s). OR’s was found for the morphology of spermatozoa (4.1 and 10.1 for medium and high exposition respectively), for the head morphology (6.1 and 4.1) and in the percentage of motile spermatozoa (9.8 and 3.5). More intensively exposed males had higher frequency of disomy in chromosomes X (p=0.012), XY (p=0.01), and Y (p<0.001). Conclusions. Bio-indicators of toxic and genetic impairment have shown lower quality of sperm in males in Teplice district.

• MeSH
• aneuploidie MeSH
• biologické markery MeSH
• dospělí MeSH
• finanční podpora výzkumu jako téma MeSH
• hybridizace in situ fluorescenční metody   MeSH
• lidé MeSH
• oxid siřičitý MeSH
• roční období MeSH
• spermie anatomie a histologie   patologie   MeSH
• znečištění ovzduší MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• přehledy MeSH
• srovnávací studie MeSH
• Geografické názvy
• Česká republika MeSH

Obesity represents a growing problem due to its impacts on human health and reproduction. In this study, we analysed semen quality, sperm DNA integrity and gene-specific CpG methylation in 116 healthy men from normal population. The men were divided into three groups according to their body mass index (BMI), and their ejaculates were analysed using standard methods, sperm chromatin structure assay (SCSA), methylation next generation sequencing (NGS) and amplicon sequencing. The sperm methylation NGS revealed six significantly differentially methylated regions (DMRs). Using subsequent targeted amplicon sequencing in 116 men, two of the DMRs were proved as differentially methylated in sperm of men with normal BMI vs. BMI ≥ 25. The DMRs were located in the EPHA8 and ANKRD11 gene. Also, we detected a significant decline in the EPHA8, ANKRD11 and CFAP46 gene methylation in association with increasing BMI values. The genes EPHA8 and ANKRD11 are involved in the nervous system and brain development; the CFAP46 gene plays a role in a flagellar assembly and is associated with sperm motility. Significantly lower rates of motile and progressive motile sperm were observed in men with BMI ≥ 30. Our results show that excess body weight can modify CpG methylation of specific genes, affect sperm motility, and compromise sperm chromatin integrity. These factors can stand behind the observed reduced fertility in men with obesity. The methylation changes might be transmitted to their offspring through sperm, and become a basis for possible developmental and reproductive issues in the next generation.

• MeSH
• analýza spermatu * MeSH
• chromatin * metabolismus   MeSH
• CpG ostrůvky MeSH
• dospělí MeSH
• index tělesné hmotnosti * MeSH
• lidé MeSH
• metylace DNA * MeSH
• mladý dospělý MeSH
• motilita spermií genetika   MeSH
• obezita genetika   MeSH
• spermie * metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Spermatozoan quality can be evaluated in different ways, here we focus on the analysis of DNA, RNA and epigenetic status of germ cells. These characterizations also can be the bases for explaining sperm quality at other levels, so we will see how some of these molecules could affect other sperm quality markers. Moreover, we consider the possibility of using some of these molecules as predictors of sperm quality in terms of the ability to produce healthy offspring. The relevant effect of different types of RNA molecules in germ line specification and spermatogenesis and the importance of germ cell DNA integrity and a proper epigenetic pattern will be also discussed. Although most studies at this level have been performed in mammals, some information is available for fish; these recent discoveries in fish models are included. We provide a general overview on how these molecules could have a deep influence in the final sperm quality.

• MeSH
• DNA genetika   MeSH
• epigeneze genetická fyziologie   MeSH
• RNA genetika   MeSH
• ryby genetika   fyziologie   MeSH
• spermatogeneze genetika   fyziologie   MeSH
• spermie fyziologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: We examined sperm quality in a cohort of city policemen in Ostrava at the end of a period with high concentrations of air pollutants (winter) and in the same cohort at the end of a relatively low exposure period (summer). METHODS: The study group was comprised of 54 nonsmoking city policemen living and working in Ostrava, Czech Republic. Average daily air-pollutant concentrations recorded by stationary monitoring for 90 days preceding the collection of semen samples were evaluated for different city districts of Ostrava. Standard semen parameters were assessed according to the guidelines of the World Health Organization (2010). The parameters were semen volume, sperm concentration, sperm morphology, sperm motility, acrosome reaction and sperm plasma membrane integrity. Sperm DNA damage was analysed by the sperm chromatin structure assay (SCSA). Sperm motion characteristics were determined by Computer Assisted Semen Analysis (CASA). RESULTS: The concentrations of all monitored pollutants (particulate matter, sulphur dioxide, nitrogen oxides, carbon monoxide, benzo[a]pyrene, benzene) were significantly increased during winter (p < 0.001), except for ozone, the concentration of which was significantly higher during summer. Sperm volume, concentration, % vitality, % sperm morphology (normal form) and % acrosome-intact sperm did not differ significantly between the monitoring periods. The percentages of total motility and progressive motility were significantly higher in March, i.e. at the end of winter (p = 0.001). However, CASA testing showed differences in sperm motion kinetics between spring and autumn samples. In the spring samples, we found a significantly lower % of straightness (p = 0.044) and the length of straight-line path (p = 0.01), while linearity and straight-line velocity were near the borderline value (p = 0.064; p = 0.054, respectively). As compared to summer, high exposure to air pollution during winter significantly increased the extent of sperm chromatin integrity damage (median 22.6 vs. 18.6%) (p = 0.003) and the proportion of immature spermatozoa (median 11.2 vs. 9.9%, p = 0.001). Sperm DNA damage negatively correlated with total motility and progressive motility (r = -0.611, -0.299; p < 0.001). The negative correlation with vitality, normal morphology and acrosome-intact sperm (r = -0.522, -0.550 and -0.511, respectively) was also significant (p < 0.001). CONCLUSION: The examination of the same cohort of city policemen at the end of a period of high air pollution and at the end of relatively low exposure reduced the effects of age, different lifestyles, different occupational exposures, localities and genetic polymorphism on sperm quality impairment associated with air pollution. This study did not demonstrate impaired standard semen parameters in association with exposure. It was shown that sperm chromatin damage and the percentage of immature sperm were highly sensitive to air pollution.

• MeSH
• analýza spermatu * MeSH
• DNA MeSH
• lidé MeSH
• motilita spermií MeSH
• sperma MeSH
• spermie MeSH
• znečištění ovzduší * statistika a číselné údaje   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Hlavní stanovisko: Vyšetření spermiogramem neposkytuje dostatek informací o kvalitě spermií. Pomocí průtokové cytometrie je možné stanovit zastoupení apoptotických spermií v ejakulátu, fragmentaci jejich DNA a integritu akrozomu. Tyto testy umožňují diagnostikovat pacienty s rizikem nižší fertility, kteří by měli být vyšetřeni andrologem. Cíl: Přehled výsledků vyšetření kvality spermií pomocí průtokové cytometrie Soubor pacientů a metody: Vyšetření ejakulátu bylo provedeno u 800 mužů (ve věku 21-66 let) léčených v Centrech asistované reprodukce GENNET. Soubor zahrnoval 366 normozoospermiků a 434 pacientů s patologickým spermiogramem. Pomocí průtokové cytometrie byl u všech pacientů stanoven počet živých a apoptotických spermií, u 213 pacientů byl stanoven počet spermií s nízkou integritou akrozomu a u 65 pacientů byl stanoven počet spermií s fragmentací DNA metodou TUNEL. Pacienti byli rozděleni do skupiny ApoHigh s vysokým zastoupením apoptotických spermií (>50 % apoptotických spermií) a ApoLow s nízkým zastoupením apoptotických spermií (<50 % apoptotických spermií). Výsledky: U normozoospermiků bylo zjištěno 16,9 % ApoHigh vzorků, u pacientů s patologickým spermiogramem 53,9 % ApoHigh vzorků. Čím více je apoptotických spermií v ejakulátu, tím vyšší je výskyt fragmentace DNA a nízké integrity akrozomu. Průměrná hodnota fragmentace DNA byla vyšší u vzorků ApoHigh než u ApoLow jak u normozoospermiků (24,0 % vs. 13,5 %), tak u pacientů s patologickým spermiogramem (33,3 % vs. 18,0 %). Vyšší průměrný počet spermií s nízkou integritou akrozomu byl zjištěn u ApoHigh vzorků než u ApoLow jak u normozoospermiků (40,2 % vs. 23,9 %), tak u pacientů s patologickým spermiogramem (53,1 % vs. 35,2 %). Závěr: Vyšší zastoupení apoptotických spermií v ejakulátu snižuje jeho kvalitu. Test apoptózy spermií, fragmentace DNA a stanovení integrity akrozomu doplňují informace získané ze spermiogramu a umožňují vytipovat rizikové subfertilní pacienty, pro které je vhodné podrobné andrologické vyšetření.

Semen analysis does not provide enough information about sperm quality. Flow cytometry tests allow us to analyse percentage of apoptotic sperm, DNA fragmentation and acrosome integrity. Sub‑fertile patients are better diagnosed using these tests. Objective: Overview of sperm quality data measured by flow cytometry. Material and method: 800 sub‑fertile men (aged 21–66 years) were treated at GENNET – assisted reproduction centres. 366 patients had normal results and 434 patients had pathological parameters of semen analysis. Flow cytometry analyses of apoptotic sperm were done in all patients. DNA fragmentation rate was measured by TUNEL assay in 65 samples. The acrosome integrity was measured in 213 samples. Two groups of patients were set according to the number of apoptotic sperm in the semen samples – ApoHigh group included samples with > 50 % of apoptotic sperm and ApoLow group included samples with < 50 % of apoptotic sperm. Results: 16.9 % of normal semen samples were ApoHigh as well as 53.9 % pathological semen. Higher average values of DNA fragmentation were detected in the ApoHigh group than in the ApoLow group both in normal semen (24.0 % vs. 13.5 %) and in pathologic semen too (33.3 % vs. 18.0 %). Higher average values of the acrosome integrity were detected in the ApoHigh group than in the ApoLow group both in normal semen (40.2 % vs. 23.9 %) and in the pathologic samples as well (53.1 % vs. 35.2 %). Conclusion: The presence of apoptotic sperm decreases the semen quality. The examination of apoptosis, DNA fragmentation and acrosome integrity together with microscopic semen analyses provide a better diagnostic tool how to identify sub‑fertile men, than simple semen analysis. These tests might be helpful for selection of patients for further uro‑andrological examination and treatment.

• MeSH
• fertilita * MeSH
• lidé MeSH
• mužská infertilita genetika   prevence a kontrola   MeSH
• průtoková cytometrie statistika a číselné údaje   MeSH
• spermie abnormality   cytologie   patologie   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• pozorovací studie MeSH

• MeSH
• fertilita fyziologie   MeSH
• finanční podpora výzkumu jako téma MeSH
• lidé MeSH
• motilita spermií fyziologie   MeSH
• počet spermií MeSH
• spermie cytologie   fyziologie   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Geografické názvy
• Česká republika MeSH

The aim of this study was to compare the spermatozoa quality parameters in spermatozoa of RAS (Recirculating Aquaculture System; RAS group) cultured (commercial pellets) and natural condition cultured (WILD group) burbot Lota lota (live prey, Pseudorasbora parva). Seven of nine fish of the RAS group produced sperm, with sperm from only four of the fish having a motility of >5%. Sperm were collected from all nine fish of the WILD group, and sperm of six of the fish from the WILD group had motility of about 100% and three had sperm with 50% to 60% motility. Spermatozoa from the RAS group had a delay in activation compared to the WILD group. Fish from the RAS group also had a lesser volume of sperm (1.8 ± 1.2 mL) collected compared to the WILD group (3.6 ± 1.2 mL). Compared to the RAS group, sperm of the WILD group had a greater proportion of saturated fatty acids (SFA), as well as the phospholipid, phosphatidylethanolamine. The findings indicate that fish grown in natural conditions may be more suitable as broodstock. Ongoing research to develop methods of enhancing reproductive performance of burbot broodstock cultured in RAS is needed to investigate whether the quality of sperm can be improved by adjusting environmental conditions, diet, or combination of these factors.

• MeSH
• analýza spermatu * MeSH
• divoká zvířata MeSH
• Gadiformes metabolismus   fyziologie   MeSH
• lipidy analýza   MeSH
• mastné kyseliny analýza   metabolismus   MeSH
• metabolismus lipidů * MeSH
• spermie chemie   metabolismus   MeSH
• vodní hospodářství metody   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

Knowledge of conditions affecting sperm quality is essential for efficient culture of fish for commercial purposes and conservation of species. Two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time of flight mass spectrometry were used to characterize the proteomic profile of Acipenser dabryanus spermatozoa relative to motility and fertilization capacity. There were differential amounts of protein in 313 spots in spermatozoa of males classified to have relatively greater or lesser spermatozoa quality. The functions of 43 of 50 selected proteins were identified. The proteins in 14 spots were involved in metabolism, and of these, proteins in 11 spots were highly abundant in spermatozoa of males categorized to have spermatozoa of greater quality, including pyruvate kinase, enolase B, phosphoglycerate kinase, lactate dehydrogenase, cytosolic malate dehydrogenase, brain creatine kinase b, Ckmb protein, and nucleoside diphosphate kinase. The proteins involved in mechanics of flagellum movement were identified, including the dynein intermediate chain, radial spoke head 1 homolog; ropporin-1-like, Bardet-Biedl syndrome 5, ADP-ribosylation factor-like protein 3, tektin-4, gamma-actin, and tubulin cytoskeleton proteins to be differentially abundant in spermatozoa that were classified relatively greater or lesser quality. Heat shock proteins, copper/zinc superoxide dismutase and peroxiredoxins, which are involved in stress response were of differential abundance in spermatozoa from males with spermatozoa in the two different classification groups. Proteins were also detected that are involved in protein folding and binding, or hydrolase activity. The results are valuable for the prediction of sperm quality and for reproduction management in A. dabryanus and other threatened species.

• MeSH
• motilita spermií MeSH
• proteomika MeSH
• rybí proteiny chemie   metabolismus   MeSH
• ryby fyziologie   MeSH
• spermie chemie   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Pro detekci molekulárních změn vyvolaných diabetes mellitus jsme použili panel protilátek proti vybraným proteinům testes a spermií. Detegovali jsme změny v morfologii a histologii varlat a změny v expresi důležitých proteinů, které jsou zapojeny do spermatogeneze a mezibuněčné komunikace. Monoklonální protilátky proti intra -akrozomálním proteinům prokázaly zhoršenou kvalitu spermií u diabetických pacientů i spermií diabetických myší. Panel vybraných protilátek lze použít k testování změn v testes a kvalitě spermií vyvolaných diabetem. Tyto výsledky jasně dokumentují vliv diabetického prostředí na morfologii a histologii varlat a kvalitu spermií.

We used a panel of antibodies against selected testicular and sperm proteins to detect molecular changes induced by diabetes mellitus. We detected changes in morphology and histology of the testes, and changes in the expression and distribution of proteins involved in spermatogenesis and intercellular communication. Monoclonal antibodies against intra -acrosomal proteins demonstrated the impaired quality of sperms from diabetic patients as well as sperms from diabetic mice. In conclusion, the panel of selected antibodies can be used to test diabetes -induced changes in the testes and sperm quality. These results clearly document the effect of the diabetic environment on the morphology and histology of the testes, and sperm quality.

• MeSH
• analýza spermatu * metody   MeSH
• diabetes mellitus 1. typu komplikace   MeSH
• fertilita * MeSH
• modely nemocí na zvířatech MeSH
• myši MeSH
• protilátky MeSH
• spermie chemie   patologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH

Chromatin remodeling, including histone post-translational modifications, during spermatogenesis can affect sperm quality and fertility, and epigenetic marks may therefore be useful for clinical evaluations of sperm. Together with histone hyperacetylation, the dimethylation of histone H3 on lysine K4 (H3K4me2) is also required during protamination. Accordingly, we evaluated the utilization of this epigenetic mark for the identification of sperm with decrease quality and immature chromatin. In this study, 99 semen samples, including 22 normozoospermic (N), 63 asthenozoospermic (A), and 14 oligoasthenozoospermic (OA) samples, were comprehensively analyzed with respect to H3K4me2 levels, DNA damage (DNA fragmentation index, DFI), and sperm immaturity (high DNA stainability, %HDS), as determined by a sperm chromatin structure assay using flow cytometry. We detected a significant relationship between H3K4me2 and %HDS (r = 0.47; p < 0.001). Furthermore, we observed negative correlations between H3K4me2 and sperm concentration, motility, and mitochondrial activity (p < 0.05). The increase in immaturity as semen quality decreased (N > A > OA) indicates the importance of chromatin immaturity and histone code deviations in sperm evaluations. Using various approaches, our study elucidated H3K4me2 as a molecular marker of sperm quality with potential use in reproductive medicine.Abbreviations: A: asthenozoospermic; AO: acridine orange; ART: assisted reproductive therapy; BWW: Biggers-Whitten Whittingham; DAPI: 4',6' -diamidino-2-phenylindole; DFI: DNA fragmentation index; H3K4me2: dimethylation of lysine K4 on histones H3; HDS: high DNA stainability; HRP: horseradish peroxidase; MACS: magnetic-activated cell sorting; N: normospermic; NGS: normal goat serum; OA: oligoasthenozoospermic; PTM: post-translational modification; SCSA: sperm chromatin structure assay; SUTI: sperm ubiquitin tag assay; TBS-T: TBS with 0.5% Tween-20.

• MeSH
• analýza spermatu MeSH
• astenozoospermie metabolismus   MeSH
• biologické markery metabolismus   MeSH
• dospělí MeSH
• histonový kód * MeSH
• histony metabolismus   MeSH
• lidé MeSH
• metylace MeSH
• oligospermie metabolismus   MeSH
• restrukturace chromatinu * MeSH
• spermie metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH