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
- analýza spermatu MeSH
- fertilizace in vitro metody MeSH
- fragmentace DNA MeSH
- habituální potrat * MeSH
- lidé MeSH
- mužská infertilita * diagnóza genetika MeSH
- senioři MeSH
- sperma MeSH
- spermie MeSH
- těhotenství MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- pozorovací studie MeSH
- MeSH
- analýza spermatu MeSH
- antibakteriální látky škodlivé účinky terapeutické užití MeSH
- fragmentace DNA MeSH
- infertilita * etiologie prevence a kontrola MeSH
- leukocyty MeSH
- lidé MeSH
- mužská infertilita etiologie prevence a kontrola MeSH
- mužské urogenitální nemoci diagnóza etiologie farmakoterapie komplikace terapie MeSH
- směrnice pro lékařskou praxi jako téma MeSH
- sperma cytologie imunologie MeSH
- spermie imunologie MeSH
- urogenitální nemoci * diagnóza etiologie farmakoterapie komplikace terapie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy 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
- apoptóza účinky léků MeSH
- bisbenzimidazol chemie MeSH
- buněčná smrt účinky léků MeSH
- buněčné jádro účinky léků metabolismus MeSH
- buněčné linie MeSH
- buňky Hep G2 MeSH
- cisplatina farmakologie MeSH
- fluorescenční mikroskopie metody MeSH
- fluorescenční spektrometrie metody MeSH
- fragmentace DNA účinky léků MeSH
- kamptothecin farmakologie MeSH
- lidé MeSH
- protinádorové látky farmakologie MeSH
- průtoková cytometrie metody MeSH
- reprodukovatelnost výsledků MeSH
- staurosporin farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem 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.
- Klíčová slova
- antispermatické protilátky,
- MeSH
- analýza spermatu * MeSH
- apoptóza MeSH
- fragmentace DNA MeSH
- infekce močového ústrojí farmakoterapie patologie MeSH
- lidé MeSH
- mužská infertilita * diagnóza MeSH
- průtoková cytometrie MeSH
- spermatogeneze MeSH
- spermie abnormality patologie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy 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 imunologie patologie virologie MeSH
- cytokiny metabolismus MeSH
- fragmentace DNA MeSH
- lidé MeSH
- lymfocyty metabolismus virologie MeSH
- oxidační stres MeSH
- reprodukční zdraví * MeSH
- SARS-CoV-2 izolace a purifikace patogenita MeSH
- spermie fyziologie virologie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy 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
- benzopyren toxicita MeSH
- embryo nesavčí účinky léků embryologie metabolismus MeSH
- embryonální vývoj účinky léků genetika MeSH
- etoposid toxicita MeSH
- fragmentace DNA účinky léků MeSH
- kamptothecin toxicita MeSH
- kometový test MeSH
- mutageny toxicita MeSH
- oprava DNA * MeSH
- poškození DNA * MeSH
- ryby embryologie genetika MeSH
- testy genotoxicity metody MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články 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
- apoptóza účinky léků MeSH
- buněčné linie MeSH
- elektrody MeSH
- fragmentace DNA MeSH
- lidé MeSH
- myši MeSH
- nanotrubičky toxicita MeSH
- nekróza chemicky indukované MeSH
- peroxidace lipidů MeSH
- reaktivní formy kyslíku metabolismus MeSH
- titan toxicita MeSH
- viabilita buněk účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Závěrečná zpráva o řešení grantu Agentury pro zdravotnický výzkum MZ ČR
Nestr.
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.
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
- MeSH
- andrologie MeSH
- asistovaná reprodukce MeSH
- diabetes mellitus 2. typu MeSH
- fragmentace DNA MeSH
- genomika MeSH
- komplikace diabetu MeSH
- modely nemocí na zvířatech MeSH
- mužská infertilita diagnóza genetika MeSH
- myši MeSH
- spermatogeneze genetika MeSH
- spermie MeSH
- vysoce účinné nukleotidové sekvenování MeSH
- Check Tag
- myši MeSH
- Konspekt
- Patologie. Klinická medicína
- NLK Obory
- genetika, lékařská genetika
- reprodukční lékařství
- diabetologie
- NLK Publikační typ
- závěrečné zprávy o řešení grantu AZV MZ ČR
Genome size and chromosome number of five Cimicidae species were compared with the similar data recently received from Cimex lectularius parasitizing human. The average nuclear DNA content (males) was 2C = 1.47 pg in C. hemipterus, 2C = 1.61 pg in C. hirundinis, 2C = 1.80 pg in C. lectularius from bats, 2C = 1.68 pg in C. pipistrelli, and 2C = 1.22 pg in Paracimex cf. chaeturus. In the genomes of all cimicid species analyzed, the average GC content ranged from 32.74% in C. pipistrelli to 35.87% in P. cf. chaeturus. Chromosome variability with two male cytotypes, 2n = 28 + X1 X2 Y and 28 + X1 X2 X3 Y, was confirmed in C. pipistrelli. In addition, intraspecific variability in chromosome number was revealed in C. lectularius from bats with 2n = 26 + X1 X2 Y and 26 + X1 X2 X3 Y. We suggest that the origin of intraspecific variability in chromosome number of C. lectularius from bats and C. pipistrelli is not only the result of simple fragmentation, but additive rearrangements like duplications are probably also involved. © 2019 International Society for Advancement of Cytometry.
- MeSH
- buněčné jádro genetika metabolismus MeSH
- Chiroptera MeSH
- chromozomy genetika MeSH
- cytogenetické vyšetření MeSH
- délka genomu MeSH
- fragmentace DNA MeSH
- gonády cytologie MeSH
- lidé MeSH
- ploidie MeSH
- pohlavní chromozomy genetika MeSH
- průtoková cytometrie MeSH
- štěnice genetika metabolismus MeSH
- zárodečné buňky chemie metabolismus MeSH
- zastoupení bazí genetika MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
A compromised detection of radiation-induced plasmid DNA fragments results in underestimation of calculated damage yields. Electrophoretic methods are easy and cheap, but they can only detect a part of the fragments, neglecting the shortest ones. These can be detected with atomic force microscopy, but at the expense of time and price. Both methods were used to investigate their capabilities to detect the DNA fragments induced by high-energetic heavy ions. The results were taken into account in calculations of radiation-induced yields of single and double strand breaks. It was estimated that the double strand break yield is twice as high when the fragments are at least partially detected with the agarose electrophoresis, compared to when they were completely omitted. Further increase by 13% was observed when the measured fragments were corrected for the fraction of the shortest fragments up to 300 base pairs, as detected with the atomic force microscopy. The effect of fragment detection on the single strand break yield was diminished.
