Cinnamtannin B-1 (CNB-1) is a naturally occurring trimeric A-type proanthocyanidin contained in several plants such as cinnamon (Cinnamomum zeylanicum). It is considered to be a potent antioxidant. The protective effect of CNB-1 against oxidative stress was assessed in red deer epididymal sperm incubated at 37 °C. Cryopreserved sperm from six stags were thawed, pooled and extended to 400 × 106 sperm/ml in BGM (bovine gamete medium). After being aliquoted, the samples were supplemented with different concentrations of CNB-1 (0, 0.1, 1, 10 and 100 μg/mL), with or without induced oxidative stress (100 μM Fe2+/ascorbate). The samples were evaluated after 0, 2 and 4 h of incubation at 37 °C. This experiment was replicated six times. Spermmotility (CASA), viability, mitochondrial membrane potential, acrosomal status, lipoperoxidation (C11 BODIPY 581/591), intracellular reactive oxygen species (ROS) production and DNA status (TUNEL) were assessed. After 4 h of incubation, CNB-1 prevented the deleterious effects of oxidative stress, thus improved sperm progressivity and velocity (P<0.05). Furthermore, 1 and 10 μM CNB-1 improved sperm linearity, even when compared to those samples that had not been subjected to oxidative stress (P<0.05). The greatest concentration, 100 μM, prevented sperm lipoperoxidation and reduced ROS production in samples subjected to oxidative stress.

• MeSH
• antioxidancia farmakologie   MeSH
• fragmentace DNA MeSH
• motilita spermií účinky léků   MeSH
• oxidační stres účinky léků   MeSH
• peroxidace lipidů MeSH
• proantokyanidiny farmakologie   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• spermie účinky léků   MeSH
• uchování spermatu veterinární   MeSH
• viabilita buněk účinky léků   MeSH
• vysoká zvěř * MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

In many fish species, sperm cryopreservation has deleterious effects and leads to a significant decrease in spermatozoa viability. However, the effect of cryopreservation on sperm cells that survive this process and are still viable is not fully understood. The objective of this study was to compare the viability and proteomes of fresh and cryopreserved sterlet (Acipenser ruthenus) sperm samples before and after live-dead cell separation using Percoll density gradient centrifugation. Both fresh and cryopreserved sperm samples were divided into two groups (with or without application of Percoll separation). At each step of the experiment, sperm quality was evaluated by video microscopy combined with integrated computer-assisted sperm analysis software and flow cytometry for live-dead sperm viability analysis. Sperm motility and the percentage of live cells were reduced in the cryopreserved group compared to the fresh group from 89% to 33% for percentage of motility and from 96% to 70% for live cells. Straight line velocity and linearity of track were significantly lower in cryopreserved samples than in those separated by Percoll before and after cryopreservation. However, the percentages of motile and live spermatozoa were higher than 90% in samples subjected to Percoll separation. Proteomic analysis of spermatozoa by two-dimensional differences in-gel electrophoresis coupled with matrix-assisted laser-desorption/ionization time-of-flight/time-of-flight mass spectrometry revealed that 20 protein spot abundances underwent significant changes in cryopreserved samples compared to fresh ones. However, only one protein spot was significantly altered when samples before and after cryopreservation followed by Percoll separation were compared. Thus, the results of this study show that cryopreservation leads to minimal proteomic changes in the spermatozoa population, retaining high motility and viability parameters. The results also suggest that global differences in protein profiles between unselected fresh and cryopreserved samples are mainly due to protein loss or changes in the lethal and sublethal damaged cell subpopulations.

• MeSH
• centrifugace - gradient hustoty metody   MeSH
• kryoprezervace metody   MeSH
• motilita spermií fyziologie   MeSH
• oxid křemičitý chemie   MeSH
• povidon chemie   MeSH
• proteomika MeSH
• ryby fyziologie   MeSH
• spermie fyziologie   MeSH
• uchování spermatu metody   MeSH
• viabilita buněk fyziologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The target of this study was to evaluate the effect of extract of the European mistletoe - Viscum album quercus L. on spermatozoa motility and viability in vitro. The CASA system was used to determine the spermatozoa motility parameters at different time intervals (0, 1, 2 and 3 h) and spermatozoa viability was determined in five different doses of Viscum album quercus L [10 (QA), 6.6 (QB), 3.3 (QC), 2.5 (QD) and 2 (QE) mg/ml]. Results in experimental groups detected a significant deterioration on rabbit spermatozoa after 1, 2 and 3 hours, compared to the control. The initial total spermatozoa motility showed increased value for all doses of Viscum album quercus in comparison to control. After in vitro culture a dose-dependent decrease (QA: reduction of 69.7 %, QB: reduction of 40.9 %) was found. For the progressive spermatozoa most significant decrease (86.8 % for QA vs. 48.5 % for QB) was detected compared to the control after 3 hours of culture. Spermatozoa viability (MTT test) was decreased in all experiment groups at the end of experiment, but the differences were not significant. Significant alterations of membrane integrity were found in groups with the highest Viscum album quercus concentration (QA, QB), but acrosome integrity showed no significant changes. Results suggest negative dose- and time-dependent effect of Viscum album quercus at higher doses on spermatozoa motility and viability parameters in vitro.

• MeSH
• časové faktory MeSH
• dub (rod) * MeSH
• králíci MeSH
• motilita spermií účinky léků   fyziologie   MeSH
• rostlinné extrakty izolace a purifikace   farmakologie   MeSH
• spermie účinky léků   fyziologie   MeSH
• viabilita buněk účinky léků   fyziologie   MeSH
• Viscum album * MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

In most teleost fishes, sperm cells are quiescent in the seminal plasma and are activated by either a drop (fresh water fish) or an increase in osmolality (marine fish) when released in the water. It is most interesting to examine how the mechanisms of sperm motility activation can adapt to a broad range of salinities, as applies to some euryhaline species, and particularly to the tilapia Sarotherodon melanotheron heudelotii, which can reproduce at salinities from 0 up to 120 in the wild. Here, the gonado-somatic index, semen characteristics, and the osmotic and ionic requirements of sperm motility activation were compared in S. m. heudelotii reared in fresh water (FW), sea water (SW), or hypersaline water (HW; salinities of 0, 35, and 70, respectively). No salinity-dependent differences were found in gonado-somatic index or semen characteristics, except for an increase of seminal plasma osmolality with increasing salinity (from 318 to 349 mOsm kg(-1) in FW and HW fish, respectively). The osmolality range allowing the highest percentages of sperm activation broadened and shifted toward higher values with increasing fish ambient salinity (150-300, 300-800, and 500-1200 mOsm kg(-1), for FW, SW, and HW fish, respectively). Nevertheless, at the three fish rearing salinities, sperm could be activated in media that were hypotonic, isotonic, or hypertonic relative to the seminal plasma, at least when some calcium was present above a threshold concentration. The [Ca(2+)] required for the activation of S. m. heudelotii sperm is (1) higher in fish reared at a higher salinity (2) higher in hypertonic than that in hypotonic activation media, whatever the fish rearing salinity, and (3) higher in the presence of Na(+) or K(+), the negative effects of which increased with an increase in fish rearing salinity. The [Ca(2+)]/[Na(+)] ​ ratios allowing for maximal sperm motility in SW or HW fish are close to those observed in natural environments, either in sea or hypersaline waters. In comparison to most teleosts with external fertilization, the total duration of sperm motility in S. m. heudelotii was exceptionally long (>2 hours regardless the fish rearing salinities). The decrease in sperm activity with increasing time since activation did not result from limiting energy reserves, as the addition of calcium in the activation medium caused most spermatozoa to become motile again. The comparison of sperm characteristics of S. m. heudelotii acclimated from FW to SW or HW with those of fish maintained all lifelong at their native salinity showed that adaptive responses were completed within 2 months or less.

• MeSH
• cichlidy fyziologie   MeSH
• motilita spermií účinky léků   MeSH
• osmotický tlak MeSH
• salinita * MeSH
• sperma fyziologie   MeSH
• spermie fyziologie   MeSH
• tolerance k soli MeSH
• viabilita buněk MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• finanční podpora výzkumu jako téma MeSH
• fluorescenční mikroskopie MeSH
• obrazová cytometrie metody   MeSH
• počet spermií metody   MeSH
• ryby fyziologie   MeSH
• spermatocyty cytologie   fyziologie   MeSH
• viabilita buněk fyziologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• srovnávací studie MeSH

The effects of air pollution on men's reproductive health can be monitored by evaluating semen quality and sperm DNA damage. We used real-time PCR to analyse the effects of air pollution on sperm mitochondrial DNA copy number (mtDNAcn) and deletion (mtDNAdel) rates in semen samples collected from 54 men in two seasons with different levels of industrial and traffic air pollution. MtDNAdel rates were significantly higher following the high exposure period and were positively correlated with mtDNAcn. However, we did not find any difference in mtDNAcn between the two seasons. MtDNAcn was positively correlated with the DNA fragmentation index and the rates of sperm with chromatin condensation defects, previously assessed by sperm chromatin structure assay, and negatively correlated with sperm concentration, progressive motility, viability, and normal morphology. This indicates that mtDNAcn is more closely associated with male fertility than mtDNAdel rates. In contrast, mtDNAdel might be a more sensitive biomarker of air pollution exposure in urban industrial environments.

• MeSH
• analýza spermatu * MeSH
• chromatin MeSH
• lidé MeSH
• mitochondriální DNA genetika   MeSH
• motilita spermií MeSH
• spermie MeSH
• variabilita počtu kopií segmentů DNA MeSH
• znečištění ovzduší * škodlivé účinky   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Cíl studie: Zhodnotit vliv rychlosti nástupu prvního buněčného dělení embryí získaných metodouICSI na úspěšnost IVF + ET.Typ studie: Prospektivní studie.Název a sídlo pracoviště: Centrum asistované reprodukce, Porodnicko-gynekologická klinika FNa LF UP, Olomouc.Metodika: U 115 cyklů IVF + ET (In Vitro Fertilizace a EmbryoTransfer) jsme za 23-26 hodin odvpichu spermie do oocytu (IntraCytosplasmic Sperm Injection, ICSI) kontrolovali rychlost nástupuprvního buněčného dělení embryí. „Early Cleavage“ embrya (EC), u kterých již proběhloprvní buněčné dělení, byla kultivována odděleně od tzv. pomalých embryí (No Early Cleavage,NEC). Po 3denní kultivaci byla transferována max. 3 embrya v 6-8buněčném stadiu. EC embryabyla transferována ve 43 cyklech, NEC embrya také ve 43 cyklech. U zbývajících 29 cyklů bylproveden tzv. kombinovaný transfer (EC + NEC embryí) a tyto cykly nebyly dále hodnoceny. Mezisoubory byly porovnávány tyto vstupní parametry: věk pacientek, počet oplozovaných oocytů,procento oplozených oocytů (fertilization rate, FR), podíl zamražených embryí, počet hodnocenýchembryí, počet zaznamenaných EC embryí a počet transferovaných embryí. Jako výstupníparametry byly hodnoceny: počet získaných těhotenství na embryotransfer (pregnancy rate, PR),počet gestačních váčků s akcí srdeční na počet přenášených embryí (implantation rate, IR), těhotenskéztráty (AB) a četnost získaných těhotenství. Statistické hodnocení bylo provedeno pomocít-testu, χ2 testu a analýzy rozptylu podle Sheffeho.Výsledky: EC embrya byla zjištěna u 57 % cyklů (n=66) po ICSI. Z celkového počtu 563 hodnocenýchembryí bylo 198 EC embryí (35,17 %). Při porovnávání vstupních parametrů jsme v cyklechs přenosem EC embryí potvrdili statisticky průkazně pouze více získaných oocytů (P

Objective: To evaluate the infl uence of early cleavage in embryos obtained using ICSI on the successrate of IVF - ET.Design: A prospective study.Setting: Centre of Assisted Reproduction, Department of Obstetrics and Gynecology, MedicalFaculty, Palacký University, Olomouc.Methods: The rate of the fi rst cleavage of embryos was checked between 23 to 26 hours after ICSIin 115 cycles IVF - ET. „Early Cleavage“ (EC) embryos which were already cleaved were cultivatedseparately from the „slow“ (No Early Cleavage, NEC) embryos. After 3 days cultivation, no morethan three 6- to 8-cell embryos were transferred. Only EC embryos were transferred in 43 cyclesand only NEC embryos were transferred also in 43 cycles. The remaining 29 cycles, where bothEC and NEC embryos had to be transferred, were not evaluated. In both groups the followingentry parameters were compared: age of the patient, number of fertilized oocytes, fertilizationrate (FR), number of frozen and evaluated embryos, number of EC embryos recorded and numberof transferred embryos. The outcome parameters compared were: pregnancy rate (PR), numberof gestational sacs with heart beats per number of transferred embryos (implantation rate, IR),abortions (AB) and the frequency of multiple pregnancy. Statistical evaluation was performed byt-test, χ2 test and by Sheffe analysis.Results: EC embryos were found in 57 % of cycles (n=66) after ICSI. Early cleavage was observedin 198 of 563 embryos (35%). Among entry parameters the only statistically signifi cant differences(P0.05) becauseof small numbers.Conclusion: The speed of the fi rst cell cleavage is a useful additional criterion for better evaluationof quality and viability of embryos.

• MeSH
• buněčné dělení MeSH
• časové faktory MeSH
• dospělí MeSH
• fertilizace in vitro metody   MeSH
• finanční podpora výzkumu jako téma MeSH
• lidé MeSH
• přenos embrya metody   MeSH
• řízení kvality MeSH
• těhotenství MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• těhotenství MeSH
• Publikační typ
• přehledy MeSH
• srovnávací studie MeSH

Persons living in industrial environments are exposed to levels of air pollution that can affect their health and fertility. The Czech capital city, Prague, and the Ostrava industrial agglomeration differ in their major sources of air pollution. In Prague, heavy traffic produces high levels of nitrogen oxides throughout the year. In the Ostrava region, an iron industry and local heating are sources of particulate matter (PM) and benzo[a]pyrene (B[a]P), especially in the winter. We evaluated the effects of air pollution on human sperm mitochondrial DNA (mtDNA). Using real-time PCR, we analysed sperm mtDNA copy number and deletion rate in Prague city policemen in two seasons (spring and autumn) and compared the results with those from Ostrava. In Prague, the sperm mtDNA deletion rate was significantly higher in autumn than in spring, which is the opposite of the results from Ostrava. The sperm mtDNA copy number did not show any seasonal differences in either of the cities; it was correlated negatively with sperm concentration, motility, and viability, and with sperm chromatin integrity (assessed with the Sperm Chromatin Structure Assay). The comparison between the two cities showed that the sperm mtDNA deletion rate in spring and the sperm mtDNA copy number in autumn were significantly lower in Prague vs. Ostrava. Our study supports the hypothesis that sperm mtDNA deletion rate is affected by the composition of air pollution. Sperm mtDNA abundance is closely associated with chromatin damage and standard semen characteristics.

• MeSH
• dospělí MeSH
• látky znečišťující vzduch toxicita   škodlivé účinky   MeSH
• lidé MeSH
• mitochondriální DNA * genetika   MeSH
• motilita spermií účinky léků   MeSH
• pevné částice toxicita   škodlivé účinky   MeSH
• policie MeSH
• roční období MeSH
• spermie * účinky léků   MeSH
• variabilita počtu kopií segmentů DNA * MeSH
• znečištění ovzduší * škodlivé účinky   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Geografické názvy
• Česká republika MeSH

• Publikační typ
• abstrakty MeSH

• Publikační typ
• abstrakt z konference MeSH