Autophagy is one of the basic cellular mechanism during preimplantation development of mammalian embryos, and it plays crucial role in several physiological processes. It is induced by interleukin (IL)-1β in mammalian cells. Our present study shows that IL-1β is important for autophagy activation in embryo development. Our in vitro culture system analysis shows effect of IL-1β in medium on the development of mouse embryos and it was found to be concentration dependent. A preimplantation embryo culture using medium containing IL-1β did not improve cleavage and blastocyst development rates of mouse embryos; however, blastocyst quality was significantly improved by increasing total cell number, especially in supplementary 20 ng/mL IL-1β. Furthermore, autophagy activation mainly occurs in 2 to 4 cell embryo and blastocyst, 20 ng/mL IL-1β into culture medium can effectively enhance levels of messenger RNA and protein of autophagy-related-factors in 2 to 4 cell embryos and blastocyst, while these factors reduce in VGX-1027 (IL-1β inhibitor) groups that also reduce the quality of blastocyst. Effects of IL-1β on the development of embryo reduced in 20 ng/mL IL-1β supplemented group when 5 mM 3-methyladenine (3-MA) was also added, which used to inhibit autophagy activation in endogenous PtdIns3Ks signal pathway. Our current results show that exogenous IL-1β can effectively induce autophagy in mouse embryos at stages of 2 to 8 cell and blastocyst, that also help to improve the quality of blastocyst.

• MeSH
• autofagie * MeSH
• blastocysta účinky léků   patologie   MeSH
• embryo savčí účinky léků   patologie   MeSH
• embryonální vývoj účinky léků   MeSH
• interleukin-1beta farmakologie   MeSH
• kultivace embrya MeSH
• myši MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

In both mitosis and meiosis, metaphase to anaphase transition requires the activity of a ubiquitin ligase known as anaphase promoting complex/cyclosome (APC/C). The activation of APC/C in metaphase is under the control of the checkpoint mechanism, called the spindle assembly checkpoint (SAC), which monitors the correct attachment of all kinetochores to the spindle. It has been shown previously in somatic cells that exposure to a small molecule inhibitor, prodrug tosyl-l-arginine methyl ester (proTAME), resulted in cell cycle arrest in metaphase, with low APC/C activity. Interestingly, some reports have also suggested that the activity of SAC is required for this arrest. We focused on the characterization of proTAME inhibition of cell cycle progression in mammalian oocytes and embryos. Our results show that mammalian oocytes and early cleavage embryos show dose-dependent metaphase arrest after exposure to proTAME. However, in comparison to the somatic cells, we show here that the proTAME-induced arrest in these cells does not require SAC activity. Our results revealed important differences between mammalian oocytes and early embryos and somatic cells in their requirements of SAC for APC/C inhibition. In comparison to the somatic cells, oocytes and embryos show much higher frequency of aneuploidy. Our results are therefore important for understanding chromosome segregation control mechanisms, which might contribute to the premature termination of development or severe developmental and mental disorders of newborns.

• MeSH
• anafázi podporující komplex metabolismus   MeSH
• embryo savčí účinky léků   metabolismus   MeSH
• embryonální vývoj účinky léků   MeSH
• kontrolní body M fáze buněčného cyklu * MeSH
• myši MeSH
• oocyty účinky léků   růst a vývoj   metabolismus   MeSH
• prekurzory léčiv MeSH
• skot MeSH
• tosylargininmethylester aplikace a dávkování   farmakologie   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• skot MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Porcine oocytes that have matured in in vitro conditions undergo the process of aging during prolonged cultivation, which is manifested by spontaneous parthenogenetic activation, lysis or fragmentation of aged oocytes. This study focused on the role of hydrogen sulfide (H2S) in the process of porcine oocyte aging. H2S is a gaseous signaling molecule and is produced endogenously by the enzymes cystathionine-β-synthase (CBS), cystathionine-γ-lyase (CSE) and 3-mercaptopyruvate sulfurtransferase (MPST). We demonstrated that H2S-producing enzymes are active in porcine oocytes and that a statistically significant decline in endogenous H2S production occurs during the first day of aging. Inhibition of these enzymes accelerates signs of aging in oocytes and significantly increases the ratio of fragmented oocytes. The presence of exogenous H2S from a donor (Na2S.9H2O) significantly suppressed the manifestations of aging, reversed the effects of inhibitors and resulted in the complete suppression of oocyte fragmentation. Cultivation of aging oocytes in the presence of H2S donor positively affected their subsequent embryonic development following parthenogenetic activation. Although no unambiguous effects of exogenous H2S on MPF and MAPK activities were detected and the intracellular mechanism underlying H2S activity remains unclear, our study clearly demonstrates the role of H2S in the regulation of porcine oocyte aging.

• MeSH
• cystathionin-beta-synthasa metabolismus   MeSH
• cystathionin-gama-lyasa metabolismus   MeSH
• embryo savčí účinky léků   MeSH
• inhibitory enzymů farmakologie   MeSH
• kultivace embrya MeSH
• kultivované buňky MeSH
• oocyty účinky léků   fyziologie   MeSH
• partenogeneze účinky léků   MeSH
• prasata MeSH
• stárnutí buněk účinky léků   MeSH
• sulfan metabolismus   farmakologie   MeSH
• sulfurtransferasy metabolismus   MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Endothelin (ET) signalling is essential for normal embryonic development. Disruption of this pathway leads to defects in the development of subsets of cranial and cephalic neural crest derivatives. Endothelin-converting enzyme 1 (ECE-1) is a ratelimiting step in the biosynthesis of ET-1. Recently, there has been considerable interest in the protective role of folic acid (FA) against congenital anomalies via increasing the expression of ET-1. We have tested whether FA supplementation can rescue craniofacial and cardiac defects observed in the ECE1-/- embryos. ECE1+/- mice were caged together to obtain litters containing embryos of all possible genotypes. The treatment group had the diet supplemented with 20 mg/kg of FA from the day of discovery of the vaginal plug. FA supplementation did not result in modified proportions of the genotypes, indicating no rescue of the embryonic mortality. There was also no effect on the litter size. Craniofacial and cardiac defects were likewise identical in the ECE1-/- embryos of both groups. There was a mild but significant reduction in the embryo size in wild-type and heterozygous FA-supplemented embryos, and there were haemorrhages in the wild-type supplemented embryos at ED14.5. Expression of ET receptor A detected by immunohistochemistry was up-regulated in the ECE1-/- embryos, but FA supplementation had no effects on the distribution of staining intensity. We conclude that FA is not able to rescue the phenotype in this model, suggesting an alternative pathway for its action. These results also caution against indiscriminate use of dietary supplements in attempts to prevent congenital anomalies.

• MeSH
• aspartátové endopeptidasy nedostatek   genetika   metabolismus   MeSH
• embryo savčí účinky léků   MeSH
• genotyp MeSH
• imunohistochemie MeSH
• kyselina listová farmakologie   MeSH
• metaloendopeptidasy nedostatek   genetika   metabolismus   MeSH
• myši knockoutované MeSH
• myši MeSH
• potravní doplňky MeSH
• regulace genové exprese enzymů účinky léků   MeSH
• těhotenství MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• asistovaná reprodukce MeSH
• embryo savčí účinky léků   MeSH
• embryonální vývoj účinky léků   MeSH
• fertilita MeSH
• infertilita * chemicky indukované   MeSH
• komplikace těhotenství * chemicky indukované   MeSH
• kouření * škodlivé účinky   MeSH
• lidé MeSH
• matka - expozice noxám MeSH
• mimoděložní těhotenství chemicky indukované   MeSH
• oocyty účinky léků   MeSH
• ovarium účinky léků   MeSH
• těhotenství účinky léků   MeSH
• Check Tag
• lidé MeSH
• těhotenství účinky léků   MeSH
• ženské pohlaví MeSH

• MeSH
• diferenciální diagnóza MeSH
• embryo savčí účinky léků   MeSH
• farmakoterapie MeSH
• komplikace těhotenství MeSH
• léčivé přípravky klasifikace   kontraindikace   MeSH
• lékové předpisy MeSH
• lidé MeSH
• nežádoucí účinky léčiv MeSH
• plod účinky léků   MeSH
• první trimestr těhotenství účinky léků   MeSH
• riziko MeSH
• těhotenství MeSH
• teratogeny farmakologie   MeSH
• Check Tag
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH

The antiproliferative and antitumor effect of wheat leaf ribonuclease was tested in vitro on the human ML-2 cell line and in vivo on athymic nude mice bearing human melanoma tumors. The antiproliferative activity of this plant ribonuclease was negligible in comparison with bovine seminal ribonuclease. In the experiments in vivo, a significant decrease of the tumor size, however, was observed in the mice treated with wheat leaf ribonuclease (27 kDa) compared with the control RNase A and polyethylene glycol. In nude mice injected intratumoraly with wheat leaf ribonuclease, the tumor size decreased from 100% in the control mice to 39% in treated mice. In the mice treated with polyethylene glycol-conjugated wheat leaf ribonuclease, the tumor reduction was observed from 100 to 28%, whereas in counterparts treated with polyethylene glycol-conjugated bovine seminal ribonuclease the tumor inhibition was reduced from 100 to 33%. Certain aspermatogenic and embryotoxic activity of wheat leaf ribonuclease and bovine seminal ribonuclease also appeared, but was lower in comparison with the effect of onconase. Mutual immunological cross-reactivity between wheat leaf ribonuclease antigens on one side and animal RNases (bovine seminal ribonuclease, RNase A, human HP-RNase and onconase) on the other side proved a certain structural similarity between animal and plant ribonucleases. Immunogenicity of wheat leaf ribonuclease was weaker in comparison with bovine seminal ribonuclease (titer of antibodies 160-320 against 1280-2560 in bovine seminal ribonuclease). Interestingly, immunosuppressive effect of wheat leaf ribonuclease tested on mixed lymphocyte culture-stimulated human lymphocytes reached the same level as that of bovine seminal RNase. The antibodies against wheat leaf ribonuclease produced in the injected mice did not inactivate the biological effect of this plant RNase in vivo. This is probably the first paper in which plant ribonuclease was used as antiproliferative and antitumor drug against animal and human normal and tumor cells and tissues in comparison with animal ribonucleases.

• MeSH
• antitumorózní látky fytogenní izolace a purifikace   terapeutické užití   toxicita   MeSH
• embryo savčí účinky léků   MeSH
• financování organizované MeSH
• imunosupresiva MeSH
• injekce intraperitoneální MeSH
• lidé MeSH
• listy rostlin chemie   MeSH
• lymfocyty imunologie   účinky léků   MeSH
• myši nahé MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádory farmakoterapie   terapie   MeSH
• polyethylenglykoly MeSH
• proliferace buněk účinky léků   MeSH
• pšenice enzymologie   MeSH
• ribonukleasy MeSH
• spermatogeneze účinky léků   MeSH
• těhotenství MeSH
• testis patologie   účinky léků   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH

Previously we have shown that monomeric RNase A has no significant biological activity, whereas its oligomers (dimer to tetramer) prepared by lyophilizing from 50% acetic acid solutions, show remarkable aspermatogenic and antitumor activities. Furthermore, conjugates prepared by chemical binding of native RNase A to polyethylene glycol (PEG) have shown a significant aspermatogenic and antitumor activities. In this work we show that the chemical conjugation of PEG to the RNase A C-dimer, and to the two RNase A trimers (NC-trimer and C- trimer) decreases the aspermatogenic activity of the oligomers while increasing their inhibitory activity on the growth of the human UB900518 amelanotic melanoma transplanted in athymic nude mice. Moreover, the PEG-conjugated RNaseA oligomers are devoid, like the free oligomers, of any embryotoxic activity.

• MeSH
• antispermatogenní látky farmakologie   MeSH
• antitumorózní látky farmakologie   chemie   MeSH
• dimerizace MeSH
• embryo savčí účinky léků   MeSH
• lidé MeSH
• melanom experimentální farmakoterapie   MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• pankreatická ribonukleasa farmakologie   chemie   MeSH
• peptidové fragmenty farmakologie   chemie   MeSH
• polyethylenglykoly farmakologie   chemie   MeSH
• spermatogeneze účinky léků   MeSH
• transplantace nádorů MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH

• MeSH
• antispermatogenní látky farmakologie   MeSH
• antitumorózní látky farmakologie   MeSH
• embryo savčí účinky léků   MeSH
• endonukleasy specifické pro jednořetězcové nukleové kyseliny * farmakologie   imunologie   MeSH
• experimentální nádory * farmakoterapie   MeSH
• fazol * enzymologie   MeSH
• lidé MeSH
• myši nahé MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• pankreatická ribonukleasa farmakologie   imunologie   MeSH
• proliferace buněk účinky léků   MeSH
• spermatogeneze účinky léků   MeSH
• spermie účinky léků   MeSH
• teratogeny farmakologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH

Kazuistika informuje o léčbě inzulínem glargin (Lantus) u 5 diabetiček 1. typu a podrobně se zabývá průběhem těhotenství u jedné z nich. Ve všech 5 případech byla léčba inzulínem glargin úspěšná a přispěla k narození zdravého dítěte. V diskusi je poukázáno na výhody, ale i možná úskalí využití inzulínových analog u gravidních diabetiček.

• Klíčová slova
• Lantus,
• MeSH
• diabetes mellitus 1. typu diagnóza   farmakoterapie   komplikace   MeSH
• embryo savčí účinky léků   MeSH
• finanční podpora výzkumu jako téma MeSH
• inzulin analogy a deriváty   terapeutické užití   MeSH
• léky s prodlouženým účinkem aplikace a dávkování   terapeutické užití   MeSH
• lidé MeSH
• těhotenství nechtěné MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• kazuistiky MeSH