Biomedical subjects

J Dostál

Publications and source records attributed to J Dostál.

At least 19 recordsLinked to original sources

PubMed2006

[Embryo quality evaluation according to the speed of the first cleavage after IntraCytoplasmic Sperm Injection (ICSI)].

OBJECTIVE: To evaluate morphological parameters of embryos obtained in the process of ICSI. DESIGN: A prospective study. SETTING: Centre of Assisted Reproduction, Dept. of Obstetrics and Gynecology, Palacký University Medical School, Olomouc. METHODS: In the present study 1116 embryos developing after ICSI (IntraCytoplasmic Sperm Injection) procedure in the period of 2001-2004 were evaluated. The beginning of the mitotic cleavage was assessed within the interval of 22-27 hours after insemination. The embryos were divided into three groups according to the speed of their division as Early Cleavage (EC) embryos, where two blastomeres were present at the time of assessment, Break Down ProNuclei stage (BDPN) where the pronuclei had already disappeared, and ProNuclei (PN) embryos, where both pronuclei were still present. In these groups the degree of fragmentation was evaluated on day two of cultivation and embryos were divided into four categories as: A--regular blastomeres, without fragmentation, B--irregular blastomeres or fragmentation below 30%, C--fragmentation 30-50%, D--fragmentation above 50%. The speed of further cleavage and average number of blastomeres were evaluated on day two and three of cultivation. Statistical analysis was preformed at the Palacky University Computer Centre. The chi2 test and t-test for independent samples were used. RESULTS: EC embryos were found in 37.4%, BDPN in 33.1% and PN in 29.5%. The degree of fragmentation between evaluated groups of embryos were statistically significant (p = 0.000). EC embryos were less fragmented (p = 0.000), had more blastomeres at the time of evaluation (p = 0.000) and their speed of cleavage was faster (p = 0.000). The cleavage of EC embryos was faster in comparison with the PN group (p = 0.000), but there were no significant differences between the EC and BDPN groups on day two of cultivation. On day three significant differences were found also between the EC and BDPN groups (p = 0.000). The embryonic developmental arrest was found only in PN embryos. CONCLUSION: The speed of the first cell cleavage is a useful additional criterion for the embryo selection for embryotransfer. EC embryos usually have better morphology and more blastomeres than the BPDN and PN ones.

Cleavage Stage, Ovum↗

PubMed2005

[Prevention of multiple pregnancy after IVF/ICSI by elective single embryo transfer--pilot study].

OBJECTIVE: To reduce the number of multiple pregnancies after IVF/ICSI treatment by an elective single embryo transfer, without significant reduction of pregnancy rate (PR). DESIGN: A pilot study. SETTING: Department of Obstetrics and Gynecology, University Hospital, Olomouc. METHODS: The patients were devided into the three groups according to number and quality of transferred embryos: elective Single EmbryoTransfer (eSET, n = 14), Single EmbryoTransfer (SET, n = 21), and Double EmbryoTransfer--control group (DET, n = 85). After 3 days of cultivation the embryos were transferred. In all groups the following parameters were compared: age of the patient, number of oocytes obtained, fertilization rate, number of frozen embryos, pregnancy rate (PR), implantation rate (IR), abortions (AB) and the number of multiple pregnancy. RESULTS: In eSET group PR and (or IR) was 42.8%, in SET group PR and (or IR) was 19%. There were no monozygotic twins in these groups. In DET group PR was 40.7% and IR 25.9%. Nine of these 33 pregnancies were twins (27%). Because of small number of the patients detailed statistical analyses were not performed. CONCLUSION: An elective transfer of one "TOP" embryo can reduce the number of multiple pregnancies without significant decrease of pregnancy rate.

Adult↗

PubMed2004

Secreted aspartate proteinases, a virulence factor of Candida spp.: occurrence among clinical isolates.

Production of secreted aspartate proteinases was determined in a set of 646 isolates of Candida and non-Candida yeast species collected from 465 patients of the University Hospital in Olomouc (Czechia) in the period 1995-2002, and Candida samples obtained from 64 healthy volunteers using solid media developed for this purpose. Using random amplified polymorphic DNA analysis (RAPD) 79 Candida isolates from blood were analyzed to show potential relationships between clustering of the fingerprints and extracellular proteolytic activity of these strains. C. albicans, C. tropicalis and C. parapsilosis possess always proteolytic activity while non-Candida species did not display any proteolysis. A tight relationship between fingerprints and extracellular proteolysis in the Candida isolates was not shown. A remarkable consistency between fingerprint clusters and proteolysis occurred in a subset of C. parapsilosis samples. Suboptimal pH of the growth medium was shown to facilitate the investigation of potential co-incidence of genotypic and phenotypic traits.

Aspartic Acid Endopeptidases↗

PubMed2004

Boar seminal immunosuppressive fraction attenuates the leptin concentration and restores the thymus mass during pregnancy in mice.

The immunosuppressive fraction (ISF) of boar seminal vesicle fluid was recently demonstrated to inhibit production of T helper (Th)1 cytokines and enhance production of Th2 cytokines. The present study shows the effect of the ISF on leptin concentrations in blood plasma and adipose tissue in mice during pregnancy. The ISF effect on thymus weight during pregnancy is also demonstrated. The leptin concentration in blood plasma and adipose tissue increased and remained high in the latter half of pregnancy. ISF treatment at the beginning of pregnancy significantly lowered the leptin concentration both in blood plasma and adipose tissue of pregnant mice. Thymus involution has been described previously in pregnant mice. ISF treatment compensated for the loss of thymus mass during the whole pregnancy in the ISF-treated mice. The treatment of pregnant mice with ISF did not affect pregnancy and litter size.

Adipose Tissue↗

PubMed2004

[Efficiency of cryoembryotransfer using embryos frozen in the pronuclear stage].

OBJECTIVE: To evaluate results of cryoembryotransfers in cycles with estrogen-progestin preparation of the endometrium. DESIGN: Prospective study. SETTING: Centre of Assisted Reproduction, Dept. of Obstetrics and Gynecology, Palacký University Medical School, Olomouc. METHODS: The study included 51 patients in whom in the period of January to August 2003 altogether 42 cycles the embryotransfer with frozen and thawed embryos were performed. The embryos for cryopreservation were obtained in the classical IVF (n = 21) or ICSI (n = 30) cycles and were cryopreserved in the pronuclear stage (PN). The storage interval varied from 3 months to 5 years. The endometrial preparation was performed in all cycles with oral estrogen--progestin substitution. Embryos in the PN stage were examined for survival (rate of intact embryos after thawing) and progress of their further development. Endometrial thickness, number of transferred embryos, pregnancy rate (PR), implantation rate (IR) and abortions (AB) were evaluated as outcome parameters. RESULTS: A total of 210 thawed embryos were evaluated. Survival was 79% without regard to the method of fertilization. For 42 cryoembryotransfers 131 embryos (62%) in 6 to 10 blastomeres after 48 hours cultivation in G 1.2 and G 2.2 fy Vitrolife were selected. Further development was more regular in embryos obtained in classical IVF cycles, where 41% of the embryos were in 4 cells stage after 24 hours of cultivation and 36% of the embryos were in 8 cells stage after 48 hours while after ICSI 32% 4 cells embryos and 26% 8 cells embryos respectively. Endometrial thickness in the day of transfer in all patients was 10.9 +/- 1.3 mm, the average number of transferred embryos was 2.4 embryos/transfer, PR was 28.5 (IVF 42.9% and ICSI 20.7%), IR 12.0% (IVF 13.0% and ICSI 11.3%) and AB 41.6% (IVF 33.3% and ICSI 50.0%). CONCLUSION: Improvement of cryopreservation procedures, freezing of embryos in the pronuclear stage and last but not least the optimal endometrial preparation can positively influence the cumulative efficiency of assisted reproduction.

Cryopreservation↗

PubMed2004

[Posthumous sperm procurement and use--ethical and legal dilemmas].

OBJECTIVE: To present a review of bioethical discussion and recommendations concerning posthumous sperm procurement and postmortem parenthood. DESIGN: Review article. SETTING: Department of Obstetrics and Gynecology, Masaryk University, Brno, Department of Obstetrics and Gynecology, Palacky University, Olomouc. SUBJECT AND METHOD: Literature search in Database of Abstracts of Reviews of the Evidence (DARE) and MEDLINEplus. Posthumous sperm procurement and cryopreservation must be performed within 36 hours after death. To established appropriate medical practice, it is important to consider all stakeholders in the decision-making process: the deceased, the requesting party, the child, the physician and the society. There are only few legislative measures concerning postmortem parenthood and posthumous sperm procurement. The essential elements for postmortem reproduction are: judicial order, ethics committee approval, bereavement period of at least 6 month before use. CONCLUSION: Posthumous sperm procurement is fraught with ethical and legal implications. All stakeholder should be considered. Society for reproductive medicine should prepare acceptable standard protocol.

Cryopreservation↗

PubMed2004

[Sonographic evaluation of endometrial preparation with oral estrogen-progesterone substitution before cryogenic embryo transfer].

OBJECTIVE: To compare the changes of sonographic features of endometrium throughout natural and artificial cycles in which oral estrogen-progesterone substitution were used in the same group of patients and discuss their clinical significance. DESIGN: A prospective study. SETTING: Dept. of Obstetrics and Gynaecology, University Hospital, Olomouc. METHODS: In 24 women with regular menstrual cycle ovulation was detected (LH, sonography) and the following sonographic features: endometrial thickness, echogenity and pulsatility index (PI) of uterine artery were evaluated in the late proliferative, early and mid secretory phase. The same sonographic examination was performed in the artificial cycle with oral estrogen-progesterone substitution used for endometrial preparation for transfer of cryopreserved-thawed embryos. RESULTS: In the late proliferative phase of the natural cycle the thickness of endometrium was higher than in the arteficial cycle (p=0.05). Triple-line echogenity prevailed in both cycles. The PI in natural cycle was lower (p=0.01) on the side of the growing Graafian follicle compared to the other side and lower (p=0.05) than in the artificial cycle. In the secretory phase there were no differences in endometrial thickness within cycles. Homogenous hyperechoic endometrium prevailed in both cycles and the lowest PI was found on the 7th day both after ovulation and after initiation of progesterone substitution. CONCLUSION: Progesterone addition after endometrial preparation with estrogen should be instituted when the endometrium thickness reached at least 9mm and isoechoic or triple line echogenity is found.

Administration, Oral↗

PubMed2003

Effects of boar seminal immunosuppressive fraction on production of cytokines by Concanavalin A-stimulated spleen cells and on proliferation of B lymphoma cell lines.

PROBLEM: The immunosuppressive fraction (ISF) of boar seminal vesicle fluid has recently been demonstrated to inhibit mitogen-stimulated proliferation of lymphocytes and antibody response to corpuscular and soluble antigens. The effects of ISF on in vitro and in vivo production of cytokines as well as its possible inhibitory effect on proliferation of B lymphoma cells remain to be elucidated. METHODS: The effect of ISF on proliferation of normal mouse spleen cells stimulated by Concanavalin A (Con A) and on mouse B lymphoma cells was measured by 3H-thymidine incorporation. Cytokines were determined in the supernatants of mouse spleen cells stimulated with Con A in the presence or absence of ISF by enzyme-linked immunosorbent assay (ELISA). In vivo cytokine production in the sera samples of mice treated with ISF and immunized with keyhole limpet hemocyanin (KLH) was followed by ELISA, too. RESULTS: We confirmed the inhibitory effect of ISF on Con A-stimulated lymphocyte proliferation. ISF affected cytokine production in the Con A-stimulated spleen cells: production of interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) was lowered, but production of IL-4, IL-6, and IL-10 was enhanced. Similarly, in the sera samples of mice immunized with keyhole limpet hemocyanin (KLH), IL-2 and IFN-gamma levels were decreased by ISF. ISF inhibited proliferation of Ag 8 and X 63-IL-2 B lymphoma cells as well. CONCLUSIONS: ISF inhibited production of T helper1 (Th1) cytokines (IL-2 and IFN-gamma) and enhanced production of Th2 cytokines (IL-4, IL-6, and IL-10). ISF seems to shift the Th1/Th2 pattern in favor of Th2. ISF exhibited an antiproliferative activity on mouse B lymphoma cells.

Adjuvants, Immunologic↗

PubMed2003

[Celiac disease and fertility disorders in women].

OBJECTIVE: To determine incidence of subclinical forms of celiac disease in women with decreased fertility. DESIGN: Screening test. SETTING: Department of Gynecology and Obstetrics, University Hospital, Palacký University, Olomouc. METHODS: 137 patients with fertility problems were included to the study. There were divided into two groups, patients with infertility and patients with repeated pregnancy loss. Screening test for celiac disease, serum level of antibodies tissue transglutaminase (tTGA) were performed in all of them. Positive test was confirmed by serological level of anti endomysium antibodies (EmA) and final diagnosis of silent celiac disease was done by enterobiopsy. RESULTS: Celiac disease was found in two infertile patients (1.67%). In one patient the silent form was diagnosed by enterobiopsy. The second disease was latent form. In patients with repeated pregnancy loss we did not observe positive screening test. CONCLUSION: We confirmed higher incidence of celiac disease in women with impaired fertility.

Celiac Disease↗

PubMed2003

[Multiple pregnancy--a significant complication of in vitro fertilization protocols].

OBJECTIVE: To evaluate the history of pregnancies after IVF-ET, to define multiple pregnancy risk factors. DESIGN: A retrospective analysis of the period of 1992-2000. SETTING: Department of Gynecology and Obstetrics, University Hospital, Palacký University, Olomouc. METHODS: 343 pregnancies were evaluated, divided into 4 groups according to the patients' ages: < or = 29 yrs. (n = 150), 30-34 yrs. (n = 122), 35-39 yrs. (n = 66), > or = 40 yrs. (n = 5). The number of transferred embryos and frequency of pregnancies in relation to the patient' ages were compared as well as the multiple pregnancy and spontaneous abortion rates. The dependence of preterm labour rate, foetus weight and the incidence of operative delivery on the frequency of pregnancies were evaluated (n = 276). The data were analysed using the chi 2 and Sheffe tests. RESULTS: Three or four transferred embryos markedly increase the number of multiple pregnancies (P < 0.01). No relation to the patient's ages was found. An increased incidence of operative deliveries (P < 0.01) as well as preterm deliveries (P < 0.05) in multiple pregnancies was found. CONCLUSIONS: The number of transferred embryos is the most important risk factor for multiple pregnancy development. It is recommended to decrease the number of transferred embryos to two or one per ET.

Abortion, Spontaneous↗

PubMed2002

Reverse effect of indomethacin on the immunosuppressive activity of boar seminal immunosuppressive fraction.

The inhibitory activity of seminal immunosuppressive fraction (ISF) on mitogen-stimulated lymphocyte proliferation and on production of antibody to a soluble antigen was modified by indomethacin or monoclonal antibody to ISF. The ability of indomethacin or monoclonal antibody to ISF to reverse the ISF-induced inhibition of mitogen-stimulated lymphocyte proliferation was estimated by measuring bromodeoxyuridine incorporation into replicated DNA. Splenocytes from mice treated with indomethacin or monoclonal antibody to ISF prior to the application of ISF were tested. The ability of indomethacin or monoclonal antibody to ISF to reverse ISF-induced suppression of antibody production was estimated by measuring antibody titers by ELISA in the blood sera from mice immunized with keyhole limpet hemocyanin (KLH). These animals were treated with indomethacin or monoclonal antibody to ISF prior to the application of ISF. The results showed that both indomethacin and monoclonal antibody to ISF reversed the inhibitory effect of ISF on mitogen-stimulated lymphocyte proliferation as well as on antibody production.Recently, we have identified ISF as a complex of the major seminal glycoproteins PSP I and PSP II. PSP II is the part that is responsible for immunosuppressive properties of the complex. To learn whether the ISF immunosuppressive effect is associated with its protein or saccharide part, we examined the deglycosylated PSP II for its antiproliferative effect on mitogen-stimulated mouse lymphocytes. The results suggest that deglycosylation of PSP II did not affect its antiproliferative activity. This suggest that PSP II immunosuppressive properties are associated with the protein and not the saccharide part of the molecule.

Animals↗

PubMed2001

Protopine hydrochloride.

Protopine hydrochloride (5,6,14,14a-tetrahydro-14a-hydroxy-7-methyl-8H-bis[1,3]benzodioxolo[5,6-a:4,5-g]quinolizinium chloride, C20H20NO5(+)-Cl(-)) is the salt of the isoquinoline alkaloid protopine. It is formed by the action of dilute hydrochloric acid on the protopine free base. The N-methyl and hydroxyl groups are in a trans configuration in the quinolizine ring and the central quinolizine N-C bond is unusually long [1.579 (2) A]. The crystal is a racemate.

Alkaloids↗

PubMed2001

Secreted aspartic proteases of Candida albicans, Candida tropicalis, Candida parapsilosis and Candida lusitaniae. Inhibition with peptidomimetic inhibitors.

The frequency of Candida infections has increased in recent years and it has been accompanied by a significant rise in morbidity and mortality. The secretion of aspartic proteases by Candida spp. was demonstrated to be one of the virulence determinants. Candida albicans is classified as the major human pathogen in the genus Candida. However, other species of this genus have been found to cause an increasing number of candidiases. We isolated secreted aspartic proteases (Saps) of C. albicans (Sap2p), C. tropicalis (Sapt1p), C. parapsilosis (Sapp1p), and C. lusitaniae (Saplp) from culture media. All the isolated proteases were N-terminally sequenced. Their specific proteolytic activities and sensitivity to series of peptidomimetic inhibitors modified in the type of scissile bond replacement as well as in the N- and C-termini were analyzed. The most divergent substrate specificity was observed for the Sap of C. tropicalis. The specificity of Sap of C. lusitaniae is most closely related to that of Sap of C. parapsilosis. We designed and prepared an inhibitor containing phenylstatine isoster that was equipotent towards all four proteases within the range of 10-10-10-9 M. The HIV-1 protease inhibitors ritonavir, saquinavir, indinavir, and nelfinavir were also tested for the inhibition of four Saps. Only ritonavir and saquinavir inhibited Sap2p, Sapt1p, Sapp1p, and Saplp in micromolar concentrations.

Amino Acid Sequence↗

PubMed2001

(-)-Argemonine hemihydrate.

(-)-Argemonine hemihydrate [systematic name: (6S,12S)-2,3,8,9-tetramethoxy-13-methyl-13-azadibenzo[b,f]bicyclo[3.3.1]nona-2,6-diene hemihydrate], C21H25NO4.0.5H2O, is a tertiary pavinane alkaloid. Both partially saturated nitrogen heterocycles adopt twisted half-chair conformations. The angle between the two aromatic rings is 86.90 (5) degrees.

Alkaloids↗

PubMed2001

[A shortened exposure of oocytes to sperm in standard in vitro fertilization].

OBJECTIVE: To find out if the shortened exposure of oocytes to sperm can influence the embryo quality and the implantation and pregnancy rate of the classical IVF also in Centre for Assisted Reproduction. DESIGN: A prospective study including 104 patients undergoing the classical IVF procedure. SETTING: Centre for Assisted Reproduction, Clinic of Gynaecology and Obstetrics, Palacký University Medical School and the University Hospital in Olomouc. METHODS: In the study group with the shortened exposure (54 patients) the oocytes were washed one hour after the insemination and transferred into the fresh medium. In the control group (50 patients) the classical procedure (e.g. transfer into the fresh medium 17 to 20 hours after insemination) has been used. The following parameters have been evaluated in both groups: the stimulation protocol, sterility cause, number of previous IVF cycles, age, serum estradiol level on the day of hCG application, number of retrieved oocytes, fertility rate (FR), embryo quality, pregnancy rate (PR) and the implantation rate (IR) expressed as the number of gestational sacs with heart action related to the number of transferred embryos. T-Test, Mann-Whitney Test and chi 2 were used for statistical evaluation. RESULTS: There were no statistically significant differences between the study and the control group in any of the parameters compared. CONCLUSION: In our study group the shortened exposure of oocytes to sperm did not influence the results of the classical IVF.

Adult↗

PubMed2001

[Ascitic fluid puncture in the treatment of severe forms of ovarian hyperstimulation syndrome].

OBJECTIVE: Evaluate the contribution and complications of ascitic fluid puncture in the treatment of severe ovarian hyperstimulation syndrome. DESIGN: Retrospective analysis of 18 patients. SETTING: Department of Gynaecology and Obstetrics, University Hospital, Palacky University, Olomouc. METHODS: 18 patients with the diagnosis of severe ovarian hyperstimulation syndrome admitted to intensive care unit from January 1996 to June 2000 were analysed. As a part of the therapy of severe OHSS 11 punctures of ascitic fluid was performed. In 3 patients the reinfusion of the ascites into the blood circulation was performed. RESULTS: The transabdominal paracentesis under the ultrasound control in 11 patients was performed. In 6 patients the puncture had to be repeated. The amount of removed ascitic fluid in single procedure was between 600-3750 ml. Total amount of fluid in different patients was between 1500-17,300 ml. Sterility of ascites and high contents of proteins was proved. In 3 patients with the refractory ascites the reinfusion of the ascitic fluid to the circulation was performed. No serious complications of the punctures were observed, 4 patients had a slight rise of body temperature, which fell down without antibiotics therapy. CONCLUSION: Puncture of the ascitic fluid is a save and effective part of the therapy of severe OHSS. In the treatment of the refractory ascites the reinfusion of the ascitic fluid should be used.

Ascites↗

PubMed2000

Inhibition of the antibody responses to rat blood transfusion antigens in mice by boar seminal immunosuppressive fraction.

PROBLEM: Immunosuppressive fraction of boar seminal vesicle fluid (ISF) was tested to muffle primary and secondary antibody responses to xenotranfusions. Contemporaneously, heparin non-binding fraction of seminal plasma (H- fraction), presumed to be identical to ISF, was used to support the results. METHOD: To study their similarity, ISF and H- fraction were analyzed by high-performance liquid chromatography and the separated proteins by N-terminal sequencing. In sera of mice treated with ISF or H- fraction, the productions of antibodies against rat erythrocytes and blood serum were evaluated by enzyme-linked immunosorbent assay (ELISA). The productions of IgM, IgA, and IgG subclasses were followed by sandwich ELISA. RESULTS: ISF and H- fraction were proved to be equal complexes of porcine seminal plasma (PSP) proteins PSP I and PSP II. Both inhibited antibody responses to rat erythrocytes and serum and the concentrations of IgM, IgG, IgG1 and IgG2 after the first transfusion with a long-lasting effect. Both suppressed the secondary antibody production if applied before the second transfusion. IgA and IgG3 stayed uninfluenced. ISF and H- fraction had an equal immunosuppressive effect. CONCLUSIONS: ISF was characterized biochemically, found to be identical to H- fraction, and determined to be powerful in overcoming unwanted exaggerated antibody responses to xenotransfusion.

Animals↗

PubMed2000

Immunolocalization of the boar seminal immunosuppressive fraction infused via uterus on the lymphocytes populating mouse genital tract tissues.

Intrauterine deposition of the immunosuppressive fraction from boar seminal vesicle fluid (ISF) led to a suppression of antibody response to soluble and corpuscular antigens in mice. By means of an immunofluorescent method using specific monoclonal antibody, ISF was detected on the membranes of white blood cells and splenocytes of mice subjected to intrauterine treatment from the third day to the thirteenth day after its deposition. ISF was also detected on the lymphocytes populating the mucosal tissues of vagina, cervix, oviduct and uterus from day 1 to 13 after its intrauterine administration. The antibody to soluble and corpuscular antigens was inhibited in the mice treated with ISF, but after the cessation of the ISF application, a normal immune response was restored within 40 days. Sandwich immunosorbent assay revealed that intrauterine infusion of ISF decreased significantly the concentration of IgG and IgM in the sera of immunized mice both after the primary and the secondary immunizations. These findings indicate that the intrauterine infusion of semen may influence the immune defense reactions and may be an important factor in the development of viral and bacterial infections of the female reproductive tract.

Adsorption↗