OBJECTIVE: Ovarian Hyperstimulation Syndrome (OHSS) is a severe health complication observed in some patients undergoing hormonal stimulation during IVF. Some authors present that OHSS is associated with Polycystic Ovary (PCO) and with a high count of growing follicles (more than 18) responding to FSH hyperstimulation, although none of them is sufficient to predict the onset of OHSS. The aim of this study is to analyze the association between PCOS and OHSS the Inhibin B-based TFF/SBM index. DESIGN: Retrospective analysis. SETTING: Assisted Reproduction Center, Department of Obstetrics and Gynecology, 1st Faculty of Medicine, Charles University and General Faculty Hospital in Prague, Sigma-Aldrich, Prague, Immunotech, a.s., Prague. METHODS: Serum and follicular fluid of 36 women (high responders with more than 18 growing follicles) in IVF program were collected at the day of oocyte pick up and used for analysis of Inhibin B. Age: mean 30.2 years (24-35 years, median 31.0 years), BMI: mean 22.34 (18.3-29.7, median 21.6). For every patient, the TFF/SBM index was calculated as follows: [concentration in FF] x [growing follicle count]/ [concentration in serum] x [body mass]. A distribution of the following parameters were compared: OHSS status, TFF/SBM index based on Inhibin B, growing follicle count and the incidence of PCOS. RESULTS: Values of the TFF/SBM index showed an association with the severe form of OHSS but not with the incidence of PCOS. CONCLUSION: These observations suggested that the incidence of PCOS is not associated with the development of severe form of OHSS, but may be still associated with a high count of growing follicles.

• MeSH
• dospělí MeSH
• fertilizace in vitro * MeSH
• folikulární tekutina chemie   MeSH
• indukce ovulace škodlivé účinky   MeSH
• inhibiny analýza   krev   MeSH
• lidé MeSH
• ovariální folikul účinky léků   patologie   MeSH
• ovariální hyperstimulační syndrom etiologie   patologie   MeSH
• syndrom polycystických ovarií komplikace   patologie   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• Názvy látek
• inhibin B MeSH Prohlížeč
• inhibiny MeSH

OBJECTIVE: Ovarian Hyperstimulation Syndrome (OHSS) is a serious complication of In Vitro Fertilisation (IVF) treatment. It is estimated that 3-5% of treated women suffer of severe form of OHSS. OHSS pathogenesis remains unknown and its treatment is only symptomatic. It is difficult to predict the risk of OHSS of an individual woman, since a current criterion--the number of punctured follicles--is not very reliable. Our aim was to find, whether in patients with a high risk of OHSS (more than 18 follicles punctured for oocyte pick-up) is possible to predict the individual risk of OHSS. As a marker we tested concentrations of Pregnancy Associated Plasma Protein-A (PAPP-A) in follicular fluid (FF) and blood serum (S). STUDY DESIGN: Prospective study. SETTINGS: Assisted Reproduction Center, Clinic of Obstetrics and Gynecology, First Faculty of Medicine, Charles University in Prague and General Teaching Hospital in Prague. METHODOLOGY: Follicular fluid and serum of 118 female patients undergoing IVF treatment was collected at the day of oocyte pick-up and analysed for PAPP-A concentration. The resulting data were then correlated with the OHSS status of individual patient. RESULTS: Mean concentration of PAPP-A in FF was 0.81 +/- 0.29 IU/l, while in S it reached only 0.0017 +/- 0.0003 IU/l. Patients with subsequent OHSS grade 1, 2 and 3 reached FF levels of PAPP-A 0.81; 0.52; and 0.73 IU/l, respectively 0.0017; 0.0017 a 0.0017 IU/l in blood serum. No correlation was found between PAPP-A FF or PAPP-A serum concentrations and the degree of OHSS. No correlation was found between PAPP-A serum concentrations and the number of follicles. CONCLUSION: 1. Pathological response on hormonal stimulation leading to OHSS is not related to the concentrations of PAPP-A in either FF or in blood serum. 2. PAPP-A does not pass from follicles to blood in a significant amount.

• MeSH
• biologické markery analýza   MeSH
• fertilizace in vitro * škodlivé účinky   MeSH
• folikulární tekutina chemie   MeSH
• lidé MeSH
• ovariální hyperstimulační syndrom diagnóza   metabolismus   MeSH
• těhotenský plazmatický protein A analýza   MeSH
• těhotenství MeSH
• Check Tag
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• Názvy látek
• biologické markery MeSH
• těhotenský plazmatický protein A MeSH

Monoclonal antibody ACR.4 recognizing specifically the 28-kDa intra-acrosomal protein was prepared by immunization of mice with acetic acid extract of boar spermatozoa, but cross-reacted also with bull intra-acrosomal protein. This monoclonal antibody was used for immunostaining analysis of bull spermatozoa before and during capacitation and ionophore-induced acrosome reaction. Immunostaining analysis showed changes of 28-kDa protein in the acrosome during capacitation and loss of this protein after induced acrosome reaction by ionophore A23187. Therefore, this monoclonal antibody can be used in the bull spermatozoa as an immunological test for detection of the acrosome state after manipulation with spermatozoa or after freezing/thawing. This test could be useful (apart from morphology and motility) for the selection of suitable spermatozoa for insemination or in vitro fertilization.

• MeSH
• akrozom chemie   imunologie   MeSH
• akrozomální reakce účinky léků   imunologie   MeSH
• calcimycin farmakologie   MeSH
• ionofory farmakologie   MeSH
• kapacitace spermií účinky léků   imunologie   MeSH
• molekulová hmotnost MeSH
• monoklonální protilátky * MeSH
• myši MeSH
• prasata MeSH
• proteiny chemie   imunologie   MeSH
• skot MeSH
• techniky in vitro MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• calcimycin MeSH
• ionofory MeSH
• monoklonální protilátky * MeSH
• proteiny MeSH

Changes of protein tyrosine phosphorylation in ejaculated boar sperm incubated in vitro were examined with the use of antiphosphotyrosine antibodies and immunoblotting. The intracellular levels of cAMP were modulated by treatment with various combinations of caffeine, 3-isobutyl-1-methylxanthine (IBMX), and dibutyryl cyclic AMP (dbcAMP), and acrosome reactions (ARs) were induced via treatment with divalent cation ionophore A23187. Proteins of Mr 34, 38, 40, and 44 (p34 ... p44) were strongly phosphorylated on tyrosine residues in freshly prepared sperm samples and at the same level during all subsequent treatments. Incubation of sperm in vitro for various periods of time induced an increase of tyrosine phosphorylation of p20, p93, and p175. The tyrosine phosphorylation of p93, p175, and several other sperm proteins was up-regulated in a concentration-dependent manner following treatment of the sperm with dbcAMP, caffeine, or IBMX alone, or with combinations of caffeine and IBMX, respectively, with dbcAMP; the tyrosine phosphorylation of p20 was not correlated with treatment of sperm with cAMP-elevating reagents. The percentage of sperm cells undergoing spontaneous ARs was not affected by the manipulation of cAMP levels and was not correlated with protein tyrosine phosphorylation. In contrast, the addition of calcium to the incubation media decreased protein tyrosine phosphorylation and elevated percentage of spontaneous ARs. The induction of ARs with A23187 caused a significant decrease of tyrosine phosphorylation of p93, p175, and p220/230, indicating that dephosphorylation on protein tyrosine residues might be associated with calcium influx during physiological ARs as well. Proteins p93 and p175 were effectively solubilized in greater than 9M urea/1% triton and in SDS sample buffer, but to only a small extent in triton, while p20 was virtually completely extractable with triton. In conjunction with the previously reported isolation of active tyrosine kinase sp42 from triton extracts of noncapacitated boar sperm cells (Berruti and Porzio, 1992: Biochim Biophys Acta 1118: 149-154), our results suggest that a cAMP-dependent event is required for tyrosine phosphorylation of triton-insoluble proteins such as p93 and p175. On the other hand, the tyrosine phosphorylation of p20 (and potentially other triton-soluble substrates) might not strictly require such cAMP up-regulation. We discuss the differences in the regulation of cAMP-dependent tyrosine phosphorylation in mouse, human, and boar sperm, and suggest that sensitivity to calcium and distinct basal levels of cyclic nucleotide PDE might correspond to species-specific reproduction strategies in mammals.

• MeSH
• AMP cyklický metabolismus   MeSH
• fosforylace MeSH
• kultivační média MeSH
• kultivované buňky MeSH
• lidé MeSH
• myši MeSH
• prasata MeSH
• signální transdukce * MeSH
• spermie metabolismus   MeSH
• tyrosin metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• AMP cyklický MeSH
• kultivační média MeSH
• tyrosin MeSH

The finding that flagellar movement in detergent-permeabilized sperm cells is restored when Mg ATP and cAMP are added implicated detergent-resistant protein kinase A (PKA) in the regulation of sperm motility. It is widely believed that only the PKA regulatory subunit RII can associate with the cytoskeleton and/or organelles. In this paper we used monoclonal antibodies against the PKA catalytic subunit and RI subunit and demonstrated that PKA type I is also associated with the sperm cytoskeleton. To our knowledge, this is the first report showing anchored PKA type I. This association was found in sperm of nonrodent mammalian species and, to a lesser extent, also in mouse sperm. The PKA catalytic subunit is bound to the cytoskeleton secondarily via its complex with the regulatory subunit. The detergent-resistant complexes of RI and catalytic subunits localize predominantly to the flagellum. Ultrastructural immunogold labeling revealed the association of detergent-resistant PKA type I with outer dense fibers (ODF) and the fibrous sheath (FS) but not with microtubules. This location is consistent with a proposed role of PKA in regulation of FS sliding on underlying ODF.

• MeSH
• detergenty farmakologie   MeSH
• lidé MeSH
• oktoxynol farmakologie   MeSH
• prasata MeSH
• proteinkinasy závislé na cyklickém AMP metabolismus   MeSH
• savci MeSH
• skot MeSH
• spermie účinky léků   enzymologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• detergenty MeSH
• oktoxynol MeSH
• proteinkinasy závislé na cyklickém AMP MeSH

Monoclonal antibodies Ds-1 and Ds-2 specifically labelling dog sperm acrosome were prepared by immunization of mice with acetic acid extracts of dog spermatozoa. Electron microscopy and indirect immunofluorescence localized the site of Ds-1 and Ds-2 proteins inside the acrosomal vesicle. Ds-1 antibody detected 55, 76, 115, 120 and 190 kDa proteins under non-reducing conditions, and 73 kDa and 54 kDa proteins after reduction (p73/Ds-1 and p54/Ds-1). 92 kDa and 40 kDa proteins recognized by Ds-2 (p92/Ds-2 and p40/Ds-2) migrated at > 200 kDa in the absence of reducing agent. In vivo, p73/Ds-1 and p54/Ds-1 are therefore likely to be present both in free and complexed form, while all of p92/Ds-2 and p40/Ds-2 form disulfide-bonded complexes. Decrease in the rate of acrosomes stained with Ds-1 and Ds-2 was correlated with the progress of capacitation resulting in the increased rate of spontaneous acrosome reactions, as suggested by a dramatic effect of A23187. Monoclonal antibody to boar acrosin (ACR-2) recognized dog sperm acrosin homologue. A higher rate of ACR-2-negative spermatozoa was observed after capacitation and A23187 treatment compared to Ds-1 and Ds-2, indicating that proteins recognized by Ds-1 and Ds-2 are localized in a specific compartment of acrosome, distinct from acrosin and possibly representing fraction of acrosomal matrix.

• MeSH
• akrosin analýza   MeSH
• akrozom chemie   účinky léků   fyziologie   MeSH
• calcimycin farmakologie   MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• fluorescenční protilátková technika nepřímá MeSH
• imunoblotting MeSH
• imunoelektronová mikroskopie MeSH
• kapacitace spermií * MeSH
• monoklonální protilátky * MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• proteiny analýza   imunologie   MeSH
• psi MeSH
• spermie ultrastruktura   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• akrosin MeSH
• calcimycin MeSH
• monoklonální protilátky * MeSH
• proteiny MeSH

Bovine cumulus-enclosed oocytes, initially cultured up to diakinesis (8 h of initial culture) or metaphase I (12 h of initial culture), were subsequently co-cultured for 6 h in contact with pig membrana granulosa (PMG) cells and then assayed for histone H1 and MAP kinase activities. In addition, the phosphorylation state of ERK 1,2 proteins was determined by Western blotting. The alterations in nuclear envelope breakdown, meiotic spindle formation and the patterns of chromosome condensation were analysed by immunofluorescence and transmission electron microscopy. The diakinesis-stage oocytes (initially cultured for 8 h) already possessed high histone H1 kinase and MAP kinase activities that were correlated with condensed and partially individualised chromosomes. The ERK 1 and most ERK 2 proteins were partly phosphorylated. Following the 6 h co-culture of these oocytes with PMG a rapid decrease in MAP kinase activity and a slower decrease in histone H1 kinase occurred, as well as ERK 1 and ERK 2 dephosphorylation. Both kinase activities and ERK 1,2 phosphorylation were fully restored following the release of the oocytes from co-culture and a subsequent culture in the absence of PMG. Moreover, the clumped bivalents were reindividualised and 56% of these oocytes reached metaphase II after 20 h of culture without PMG. The metaphase I oocytes, initially cultured for 12 h, displayed a fusiform meiotic spindle and a metaphase array of chromosomal bivalents, accompanied by high levels of both histone H1 and MAP kinase activity. Co-culture of MI oocytes with PMG abolished the activity of both kinases and caused the dephosphorylation of ERK 1 and ERK 2. Furthermore, the spindle microtubules were depolymerised and the chromosomal bivalents clumped into a single mass. Neither of the protein kinase activities nor the meiotic spindle were restored following subsequent culture in the absence of PMG for up to 20 h. These observations indicate that under in vitro conditions membrana granulosa cells can cause a prompt decrease in histone H1 and MAP kinase activities, and metaphase I oocytes. While these events are fully reversible in late diakinesis oocytes, metaphase I oocytes did not complete maturation after release from co-culture.

• MeSH
• chromozomy MeSH
• folikulární buňky fyziologie   MeSH
• fosforylace MeSH
• histonkinasa metabolismus   MeSH
• jaderný obal metabolismus   MeSH
• kokultivační techniky MeSH
• meióza fyziologie   MeSH
• metafáze fyziologie   MeSH
• mikrotubuly MeSH
• mitogenem aktivovaná proteinkinasa 1 MeSH
• mitogenem aktivovaná proteinkinasa 3 MeSH
• mitogenem aktivované proteinkinasy * MeSH
• oocyty cytologie   enzymologie   fyziologie   MeSH
• prasata MeSH
• proteinkinasa CDC2 metabolismus   MeSH
• proteinkinasy závislé na vápníku a kalmodulinu metabolismus   MeSH
• skot MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• histonkinasa MeSH
• mitogenem aktivovaná proteinkinasa 1 MeSH
• mitogenem aktivovaná proteinkinasa 3 MeSH
• mitogenem aktivované proteinkinasy * MeSH
• proteinkinasa CDC2 MeSH
• proteinkinasy závislé na vápníku a kalmodulinu MeSH

In this study we used previously characterized monoclonal antibodies to acrosin (ACR.2) and to an acrosomal matrix antigen (ACR.4) to analyze the acrosin-binding activity of a 28-kDa putative acrosin-binding protein from the acrosomal matrix. The 28-kDa protein bound proacrosin and the 49-kDa form of acrosin (alpha-acrosin) but it did not bind the 36-kDa acrosin form (beta-acrosin). The acrosin-binding activity of the 28-kDa protein was stimulated by Ca2+, inhibited by Mg2+, and removed by disulphide bond reduction. Induction of the acrosome reaction by a calcium ionophore resulted in proteolytic cleavage of the 28-kDa protein, giving rise to a 12-kDa degradation product that was the only form of ACR.4 antigen released to incubation media; the release of the ACR.4 antigen was closely correlated with that of acrosin. The release of alpha-acrosin to incubation media was accelerated in the presence of ACR.4 antibody. In a cell-free system, a limited cleavage of the purified 28-kDa protein into immunoreactive degradation products was catalyzed by acrosin but not by trypsin or chymotrypsin. The data suggest that the 28-kDa acrosomal protein helps to maintain acrosomal matrix integrity and controls the acrosin release from acrosome-reacted cells.

• MeSH
• akrosin metabolismus   MeSH
• akrozom imunologie   fyziologie   MeSH
• antigeny metabolismus   MeSH
• calcimycin farmakologie   MeSH
• chymotrypsin metabolismus   MeSH
• kapacitace spermií MeSH
• kationty dvojmocné MeSH
• molekulová hmotnost MeSH
• monoklonální protilátky MeSH
• prasata MeSH
• prekurzory enzymů metabolismus   MeSH
• rozpustnost MeSH
• spermie účinky léků   fyziologie   MeSH
• transportní proteiny chemie   metabolismus   MeSH
• trypsin metabolismus   MeSH
• vápník farmakologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• akrosin MeSH
• antigeny MeSH
• calcimycin MeSH
• chymotrypsin MeSH
• kationty dvojmocné MeSH
• monoklonální protilátky MeSH
• prekurzory enzymů MeSH
• proacrosin MeSH Prohlížeč
• transportní proteiny MeSH
• trypsin MeSH
• vápník MeSH

In this study we used a previously characterized monoclonal antibody to analyze the molecular conversions of acrosin during the acrosomal exocytosis induced by ionophore A23187. Before sperm exposure to the ionophore, most of the sperm acrosin was in the form of proacrosin (55-kDa and 53-kDa forms). Upon exposure to the ionophore, the concentration of proacrosin in sperm samples decreased rapidly and was negatively correlated with the progression of exocytosis. After 1 h of ionophore treatment, proacrosin was quantitatively converted into the two active acrosin forms, alpha-acrosin (49 kDa) and beta-acrosin (36 kDa). However, products of further acrosin conversions were not found after this treatment. As compared with the speed of acrosin activation during sperm contact with the ionophore, the ionophore-induced release of acrosin from the sperm cells into the soluble fraction was apparently delayed, and only the active acrosin forms (49 kDa and 36 kDa) were found in sperm incubation media. External Ca2+ influenced the speed of proacrosin conversion in a concentration-dependent manner. The ionophore-induced activation of proacrosin and acrosome reaction were partially inhibited by trypsin inhibitors. The results suggest that proacrosin activation is an essential step in the mechanism of the acrosomal exocytosis.

• MeSH
• akrosin chemie   MeSH
• calcimycin farmakologie   MeSH
• exocytóza účinky léků   MeSH
• inhibitory trypsinu farmakologie   MeSH
• prasata MeSH
• prekurzory enzymů chemie   MeSH
• spermie účinky léků   enzymologie   MeSH
• vápník farmakologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• akrosin MeSH
• calcimycin MeSH
• inhibitory trypsinu MeSH
• prekurzory enzymů MeSH
• proacrosin MeSH Prohlížeč
• vápník MeSH

Sperm coating proteins of 16, 17, and 19 kDa have been purified from boar seminal plasma. The 17 kDa protein has been identified as an antigen recognized by monoclonal antibody ACR.3 and is thus identical to low molecular mass zona pellucida binding protein from boar spermatozoa (Moos et al., 1990). The 17 and 19 kDa proteins are glycosylated and tend to form hetero-complexes. The 17 kDa ACR.3 antigen is sequentially released from the sperm cell surface during capacitation and, after induction of the acrosome reaction, the 16 kDa form was also observed. Immunocytochemical studies on boar reproductive tissues have suggested that the seminal vesicle epithelium may be the source of these proteins.

• MeSH
• akrozom fyziologie   MeSH
• antigeny povrchové imunologie   izolace a purifikace   MeSH
• ejakulace MeSH
• epididymis MeSH
• kapacitace spermií MeSH
• monoklonální protilátky imunologie   MeSH
• myši MeSH
• orgánová specificita MeSH
• prasata metabolismus   MeSH
• sperma chemie   MeSH
• zona pellucida metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigeny povrchové MeSH
• monoklonální protilátky MeSH