The use of microfluidic sperm sorting (MFSS) systems in infertility treatment is increasing due to their practicality and ease of use. While often presented as highly effective, their efficacy in patients with varying sperm analysis results remains uncertain. In this study, we evaluated the effectiveness of MFSS compared with the swim-up (SU) technique in terms of oxygen radical levels and spermiogram parameters. Samples from each patient were processed using both methods, followed by assessments of sperm concentration, motility, morphology, DNA integrity, acrosomal status, and mitochondrial membrane potential. Participants were selected based on sperm analysis and categorized as normozoospermic (n = 40) or non-normozoospermic (n = 28). An analysis of separation techniques revealed no significant differences, except for a lower percentage of DNA-fragmented sperm in the MFSS group compared with SU within the non-normozoospermic cohort (SU: 10.0% vs. MFSS: 5.69%, p = 0.027). No differences were observed between SU and MFSS in normozoospermic men. The MFSS method is a simple technique, frequently used in laboratories, that yields good results but does not offer a substantial advantage over SU. The primary benefit of MFSS appears to be a significant reduction in the proportion of sperm with DNA fragmentation compared with SU in patients with abnormal sperm analysis results.

• MeSH
• Semen Analysis methods   MeSH
• Adult MeSH
• DNA Fragmentation MeSH
• Sperm Injections, Intracytoplasmic * methods   MeSH
• Humans MeSH
• Membrane Potential, Mitochondrial MeSH
• Microfluidics * methods   MeSH
• Sperm Motility * MeSH
• Infertility, Male therapy   MeSH
• Cell Separation * methods   MeSH
• Spermatozoa * cytology   metabolism   MeSH
• Check Tag
• Adult MeSH
• Humans MeSH
• Male MeSH
• Publication type
• Journal Article MeSH
• Comparative Study MeSH

Mužská neplodnost je komplexní stav s mnoha etiologickými faktory, včetně hormonálních, anatomických, genetických a vlivů životního stylu. Přestože se mužský faktor podílí až na 40 % případů neplodnosti, přesná příčina zůstává často neznámá (tzv. idiopatická neplodnost). Současné diagnostické metody zahrnují detailní klinické vyšetření, analýzu ejakulátu, hormonální profil, genetické testování a zobrazovací techniky. Výrazný pokrok byl zaznamenán v oblasti testování fragmentace DNA spermií a měření oxidačního stresu, jež poskytují širší pohled na kvalitu spermií. Terapeutické postupy se liší podle příčiny neplodnosti – od farmakologické léčby a chirurgických intervencí až po asistovanou reprodukci. Rychle se rozvíjející výzkum v oblasti regenerativní a genové terapie slibuje nové možnosti léčby. Nedílnou součástí prevence i léčby je rovněž zdravý životní styl a management rizikových faktorů.

Male infertility is a multifactorial condition influenced by hormonal, anatomical, genetic, and lifestyle factors. Although the male factor contributes to up to 40% of infertility cases, the exact etiology often remains unknown (so-called idiopathic infertility). Current diagnostic approaches include detailed clinical evaluation, semen analysis, hormonal assessment, genetic testing, and imaging techniques. Significant progress has been achieved in sperm DNA fragmentation testing and oxidative stress evaluation, offering a broader insight into sperm quality. Treatment strategies vary depending on the underlying cause, ranging from pharmacological therapy and surgical interventions to assisted reproductive technologies. Rapid advances in regenerative medicine and gene therapy show promise for novel therapeutic options. A healthy lifestyle and risk factor management are integral to both prevention and treatment.

• MeSH
• Semen Analysis MeSH
• Andrology trends   MeSH
• Reproductive Techniques, Assisted MeSH
• Diagnostic Techniques and Procedures classification   MeSH
• DNA Fragmentation MeSH
• Humans MeSH
• Infertility, Male * diagnosis   etiology   therapy   MeSH
• Oxidative Stress MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Publication type
• Review MeSH

Role of male factor in recurrent abortion and in vitro fertilization failure has not been fully defined yet and there is much controversy about evaluating male patients with normal semen analysis. One of the factors that might help establish the male role is DNA fragmentation index. However, strong correlation between this factor and quality of semen, has caused many clinicians to believe that it does not help in abortion and implantation failure. We aim to assess this factor in our patients. In a prospective observational study, we assessed age, duration of infertility, undesired fertility related events (assisted reproductive techniques attempts and abortions), semen parameters and DNA fragmentation index in patients with multiple abortions or in vitro fertilization failures and analysed the results by statistical software SPSS version 24. DNA fragmentation index was remarkably correlated with age, duration of infertility and semen parameters. Among all groups in our study, patients with abnormal semen analysis had statistically significant higher level of DNA fragmentation. Ten percent of patients with normal or slightly abnormal semen analysis had abnormally high SDFI (sperm DNA fragmentation index). Checking DNA fragmentation index is recommended in all couples with fertilization problems even in the presence of normal semen analysis. It might be more reasonable to assess it in aged men, long duration of infertility or candidates with remarkable semen abnormality.

• MeSH
• Semen Analysis MeSH
• Fertilization in Vitro methods   MeSH
• DNA Fragmentation MeSH
• Abortion, Habitual * MeSH
• Humans MeSH
• Infertility, Male * diagnosis   genetics   MeSH
• Aged MeSH
• Semen MeSH
• Spermatozoa MeSH
• Pregnancy MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Aged MeSH
• Pregnancy MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Observational Study MeSH

• MeSH
• Semen Analysis MeSH
• Anti-Bacterial Agents adverse effects   therapeutic use   MeSH
• DNA Fragmentation MeSH
• Infertility * etiology   prevention & control   MeSH
• Leukocytes MeSH
• Humans MeSH
• Infertility, Male etiology   prevention & control   MeSH
• Male Urogenital Diseases diagnosis   etiology   drug therapy   complications   therapy   MeSH
• Practice Guidelines as Topic MeSH
• Semen cytology   immunology   MeSH
• Spermatozoa immunology   MeSH
• Urogenital Diseases * diagnosis   etiology   drug therapy   complications   therapy   MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Publication type
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH

At present, nuclear condensation and fragmentation have been estimated also using Hoechst probes in fluorescence microscopy and flow cytometry. However, none of the methods used the Hoechst probes for quantitative spectrofluorometric assessment. Therefore, the aim of the present study was to develop a spectrofluorometric assay for detection of nuclear condensation and fragmentation in the intact cells. We used human hepatoma HepG2 and renal HK-2 cells cultured in 96-well plates treated with potent apoptotic inducers (i.e. cisplatin, staurosporine, camptothecin) for 6-48 h. Afterwards, the cells were incubated with Hoechst 33258 (2 µg/mL) and the increase of fluorescence after binding of the dye to DNA was measured. The developed spectrofluorometric assay was capable to detect nuclear changes caused by all tested apoptotic inducers. Then, we compared the outcomes of the spectrofluorometric assay with other methods detecting cell impairment and apoptosis (i.e. WST-1 and glutathione tests, TUNEL, DNA ladder, caspase activity, PARP-1 and JNKs expressions). We found that our developed spectrofluorometric assay provided results of the same sensitivity as the TUNEL assay but with the advantages of being fast processing, low-cost and a high throughput. Because nuclear condensation and fragmentation can be typical markers of cell death, especially in apoptosis, we suppose that the spectrofluorometric assay could become a routinely used method for characterizing cell death processes.

• MeSH
• Apoptosis drug effects   MeSH
• Bisbenzimidazole chemistry   MeSH
• Cell Death drug effects   MeSH
• Cell Nucleus drug effects   metabolism   MeSH
• Cell Line MeSH
• Hep G2 Cells MeSH
• Cisplatin pharmacology   MeSH
• Microscopy, Fluorescence methods   MeSH
• Spectrometry, Fluorescence methods   MeSH
• DNA Fragmentation drug effects   MeSH
• Camptothecin pharmacology   MeSH
• Humans MeSH
• Antineoplastic Agents pharmacology   MeSH
• Flow Cytometry methods   MeSH
• Reproducibility of Results MeSH
• Staurosporine pharmacology   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

S neplodností, tedy s neschopností otěhotnět v průběhu jednoho roku pravidelného nechráněného pohlavního styku, se potýká cca 15 % párů ve vyspělých zemích. Jelikož se jedná o diagnózu konkrétního páru, a nikoli jedince, je namístě přistupovat k páru jako celku, ne odděleně k ženě či muži. Pro posouzení fertilního potenciálu muže je za zlatý standard již několik desetiletí považován spermiogram, tedy nativní vyšetření ejakulátu. Spermiogram je ve své základní podobě pouze morfologickým, nikoli funkčním vyšetřením, proto nemusí vždy spolehlivě informovat o skutečném fertilním potenciálu daného muže. Z tohoto důvodu jsou do praxe zaváděny nové metody pro zlepšení diagnostiky a následné terapie. Uvádíme možnosti vyšetření ejakulátu průtokovou cytometrií a vliv asymptomatických urogenitálních infekcí na plodnost.

In developed countries, approximately 15% of couples suffer from infertility, i.e. they do not conceive within one year of a regular unprotected sexual intercourse. Since infertility is the only one diagnosis of a couple, and not of an individual, it is essential to examine the couple as the unit. Sperm analysis, i.e. native microscopic evaluation, has been used for decades as a golden standard for male fertile potential assessment. Sperm analysis, in its fundamental form, has been only morphological, and not functional evaluation of ejaculate, thus it might not give us reliable information about actual fertile potential of an individual male. On that account, new methods are being introduced to the clinical practice with a goal to improve diagnostics and subsequent treatment. The article presents these new methods, namely flow cytometry, and the impact of asymptomatic urogenital infections on fertility.

• Keywords
• antispermatické protilátky,
• MeSH
• Semen Analysis * MeSH
• Apoptosis MeSH
• DNA Fragmentation MeSH
• Urinary Tract Infections drug therapy   pathology   MeSH
• Humans MeSH
• Infertility, Male * diagnosis   MeSH
• Flow Cytometry MeSH
• Spermatogenesis MeSH
• Spermatozoa abnormalities   pathology   MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Publication type
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH

Coronavirus disease 2019 (COVID-19) has emerged as a new public health crisis, threatening almost all aspects of human life. Originating in bats, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is transmitted to humans through unknown intermediate hosts, where it is primarily known to cause pneumonia-like complications in the respiratory system. Organ-to-organ transmission has not been ruled out, thereby raising the possibility of the impact of SARS-CoV-2 infection on multiple organ systems. The male reproductive system has been hypothesized to be a potential target of SARS-CoV-2 infection, which is supported by some preliminary evidence. This may pose a global threat to male fertility potential, as men are more prone to SARS-CoV-2 infection than women, especially those of reproductive age. Preliminary reports have also indicated the possibility of sexual transmission of SARS-CoV-2. It may cause severe complications in infected couples. This review focuses on the pathophysiology of potential SARS-CoV-2 infection in the reproductive organs of males along with their invasion mechanisms. The risks of COVID-19 on male fertility as well as the differences in vulnerability to SARS-CoV-2 infection compared with females have also been highlighted.

• MeSH
• COVID-19 immunology   pathology   virology   MeSH
• Cytokines metabolism   MeSH
• DNA Fragmentation MeSH
• Humans MeSH
• Lymphocytes metabolism   virology   MeSH
• Oxidative Stress MeSH
• Reproductive Health * MeSH
• SARS-CoV-2 isolation & purification   pathogenicity   MeSH
• Spermatozoa physiology   virology   MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Publication type
• Journal Article MeSH
• Review MeSH

DNA damage caused by exogenous or endogenous factors is a common challenge for developing fish embryos. DNA damage repair (DDR) pathways help organisms minimize adverse effects of DNA alterations. In terms of DNA repair mechanisms, sturgeons represent a particularly interesting model due to their exceptional genome plasticity. Sterlet (Acipenser ruthenus) is a relatively small species of sturgeon. The goal of this study was to assess the sensitivity of sterlet embryos to model genotoxicants (camptothecin, etoposide, and benzo[a]pyrene), and to assess DDR responses. We assessed the effects of genotoxicants on embryo survival, hatching rate, DNA fragmentation, gene expression, and phosphorylation of H2AX and ATM kinase. Exposure of sterlet embryos to 1 µM benzo[a]pyrene induced low levels of DNA damage accompanied by ATM phosphorylation and xpc gene expression. Conversely, 20 µM etoposide exposure induced DNA damage without activation of known DDR pathways. Effects of 10 nM camptothecin on embryo development were stage-specific, with early stages, before gastrulation, being most sensitive. Overall, this study provides foundational information for future investigation of sterlet DDR pathways.

• MeSH
• Benzo(a)pyrene toxicity   MeSH
• Embryo, Nonmammalian drug effects   embryology   metabolism   MeSH
• Embryonic Development drug effects   genetics   MeSH
• Etoposide toxicity   MeSH
• DNA Fragmentation drug effects   MeSH
• Camptothecin toxicity   MeSH
• Comet Assay MeSH
• Mutagens toxicity   MeSH
• DNA Repair * MeSH
• DNA Damage * MeSH
• Fishes embryology   genetics   MeSH
• Mutagenicity Tests methods   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

The present study reports on a comprehensive investigation of mechanisms of in vitro cytotoxicity of high aspect ratio (HAR) bundles formed from anodic TiO2 nanotube (TNT) layers. Comparative cytotoxicity studies were performed using two types of HAR TNTs (diameter of ∼110 nm), differing in initial thickness of the nanotubular layer (∼35 μm for TNTs-1 vs. ∼10 μm for TNTs-2). Using two types of epithelial cell lines (MDA-MB-231, HEK-293), it was found that nanotoxicity is highly cell-type dependent and plausibly associates with higher membrane fluidity and decreased rigidity of cancer cells enabling penetration of TNTs to the cell membrane towards disruption of membrane integrity and reorganization of cytoskeletal network. Upon penetration, TNTs dysregulated redox homeostasis followed by DNA fragmentation and apoptotic/necrotic cell death. Both TNTs exhibited haemolytic activity and rapidly activated polarization of RAW 264.7 macrophages. Throughout the whole study, TNTs-2 possessing a lower aspect ratio manifested significantly higher cytotoxic effects. Taken together, this is the first report comprehensively investigating the mechanisms underlying the nanotoxicity of bundles formed from self-organised 1-D anodic TNT layers. Except for description of nanotoxicity of industrially-interesting nanomaterials, the delineation of the nanotoxicity paradigm in cancer cells could serve as solid basis for future efforts in rational engineering of TNTs towards selective anticancer nanomedicine.

• MeSH
• Apoptosis drug effects   MeSH
• Cell Line MeSH
• Electrodes MeSH
• DNA Fragmentation MeSH
• Humans MeSH
• Mice MeSH
• Nanotubes toxicity   MeSH
• Necrosis chemically induced   MeSH
• Lipid Peroxidation MeSH
• Reactive Oxygen Species metabolism   MeSH
• Titanium toxicity   MeSH
• Cell Survival drug effects   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Celkově se zvyšující podíl neplodných párů je v současné době závažným problémem. Mužský faktor je příčinou v přibližně 60% a tento podíl bude narůstat. Jedním z rizikových faktorů u mužské neplodnosti je i diabetes mellitus (DM), který se projevuje jako persistentní hyperglykémie u obou svých typů. Nárůst četnosti DM a snižující věk diagnózy se negativně kombinuje se zvyšujícím se věkem reprodukce. Epidemiologické studie prokazují asociaci persistentní hyperglykémie se sníženou kvalitou spermií a zvířecí modely přinesly první data ohledně jejího dopadu na spermatogenezi. Dosud byl dopad DM při vyhodnocování mužské infertility opomíjen a plný rozsah molekulárně patogenetických změn u DM na lidskou spermatogenezi nebyl probádaný. Tento projekt je proto zaměřen na: 1) zhodnocení vlivu persistentní hyperglykémie u idiopatické mužské neplodnosti a při úspěšnosti asistované reprodukce (ART) u DM, 2) výběr vhodné andrologické metody při ART u DM, a 3) objasnění epigenetických a molekulárních změn při spermatogenezi indukovaných persistentní hyperglykémií na zvířecím modelu; The increasing frequency of infertility has become a major public health concern. Male infertility is the primary or contributing cause in approximately 60% of all cases. Diabetes mellitus (DM) reflected by persistent hyperglycaemia (PH) is suspected to be a contributing factor in male infertility. Increase in DM frequency together with its lower age at diagnosis is negatively compounded by the increasing age at reproduction. Epidemiological data together with analyses in animal models provided evidence of the association of PH on decreased sperm quality and impaired spermatogenesis. The role of PH in male infertility has been under-diagnosed thus far, while the exact molecular pathogenesis related to PH has not been studied in human subjects. This project therefore focuses on the 1) pathogenetic role of PH in male infertility and success in ART in these patients, 2) selection of an optimal andrologic dg. method in such instances, and 3) elucidation of the mol. pathogenesis and epigenetic disturbances due to PH in an animal model.

• MeSH
• Andrology MeSH
• Reproductive Techniques, Assisted MeSH
• Diabetes Mellitus, Type 2 MeSH
• DNA Fragmentation MeSH
• Genomics MeSH
• Diabetes Complications MeSH
• Disease Models, Animal MeSH
• Infertility, Male diagnosis   genetics   MeSH
• Mice MeSH
• Spermatogenesis genetics   MeSH
• Spermatozoa MeSH
• High-Throughput Nucleotide Sequencing MeSH
• Check Tag
• Mice MeSH

• Conspectus
• Patologie. Klinická medicína
• NML Fields
• genetika, lékařská genetika
• reprodukční lékařství
• diabetologie
• NML Publication type
• závěrečné zprávy o řešení grantu AZV MZ ČR