Does EmbryoGlue transfer medium affect embryo transfer success rate?

OBJECTIVE: Verification of the effect of EmbryoGlue (EG) transfer medium enriched with hyaluronan on the embryo transfer success rate. DESIGN: A prospective study. SETTING: Fertimed, Fertility Treatment Centre, Olomouc. METHODS: 484 patients undergoing IVF + ET were engaged who were divided according to the medium used during transfer: Sage or Vitrolife culture medium (n = 135) and EG transfer medium (n = 349). The influence of other factors was also evaluated: age, number of received oocytes, percentage of fertilised oocytes, endometrium height on the day of transfer, number of preserved embryos, number of transferred embryos and the length of cultivation (48, 72, 96, 120 h). These factors were considered quantitative variables while the method of hormone stimulation, the type of culture medium and the application of EG were considered qualitative (categorical) variables. RESULTS: With the use of EG the chances of conception increased by approximately 9%. The negative effect of higher age on implantation proportion and the positive effect of cultivation prolongation were proven. The effect of EG on miscarriages was not proven. The probability of miscarriage diminished with higher oocyte fertilisation. The culture medium used (Sage or Vitrolife) before the application of EG does not affect the gravidity result. CONCLUSION: Using the transfer medium of EmbryoGlue before embryo transfer affects the pregnancy rate (PR) in a positive manner. The influence on pregnancy loss with the application of EG was not proven.

HCG level after embryo transfer as a prognostic indicator of pregnancy finished with delivery.

OBJECTIVE: To examine patients after embryo transfer for predictive influence of the human chorionic gonadotropin (hCG) level on the probability of finishing pregnancy with delivery. DESIGN: Retrospective study. SETTING: Fertimed, Fertility Treatment Centre, Olomouc. METHODS: 490 patients pregnant after IVF + ET treat-ment placed in the study. The influence of other factors: age (patients 20-50), number of transferred embryos (one, two and more) and the length of cultivation on the probability of finishing pregnancy with delivery or loss and the possibility to predict multiple pregnancy was also related to the known hCG value. The transferred embryos were divided into two groups - early (cultivation 48 and 72 hrs) and prolonged cultivation (PC 96 and 120 hrs). The answer here, therefore, is a multinomial variable with four levels. For that reason the data was analysed through a multinomial logistic model vs. multinomial distribution of a mistake and generalised logistic link function. RESULTS: The hCG level grows exponentially in the course of the 9th to 17th day after an embryo transfer (ET). The probability of one child delivery after the transfer of one embryo overdue after prolonged cultivation (96 or 120 hrs) grows with the average and above-average hCG values on the day of the draw. The hCG value was 678 (564-815) IU/l1 on the 14th day after ET in pregnancy ended in delivery, 321 (216-477) IU/l on average in abortion, 82 (51-132) IU/l in biochemical pregnancy and 1070 (737-1554) IU/l in multiple pregnancy. The probability of multiple pregnancy increased with hCG values greatly above the average and on the other hand, below-average values indicated abortion or biochemical pregnancy. The patients age was not proven to be of significant influence, the hCG level slightly decreased with higher age. On the contrary, an increasing frequency of abortions depending on the increasing age of the mother was once again confirmed. CONCLUSION: The measured hCG values are considerably different depending on the pregnancy result, which is why this value is considered a quality predictive factor of the pregnancy result.

[How is the Czech oocyte donor? The analysis of their profile]. / Jaká je ceská dárkyne oocytu?Analýza profilu.

OBJECTIVE: The aim of our study was to investigate (a) the basic socio-demographic characteristics of Czech oocytes donors, (b) their attitudes towards anonymous or nonanonymous donation and (c) their motivations to donate oocytes. DESIGN: Original article. SETTING: Clinic of Reproductive Medicine and Gynecology Zlin. METHODS: A total of 215 oocytes donors participated in this study, which was undertaken in years 2015-2016. The data were obtained by an anonymous questionnaire. Descriptive and inferential statistics were used to analyzed the data. CONCLUSION: The results show that (a) mean age of oocytes donors was 26 years. Single women (60%) with secondary level education (57%) prevailed. More than half of them (55%) had at least one child on their own. About 39% of donors were fully employed, 29% women on maternity leave and 16% students. (b) For the majority of participants the anonymity is very important. Almost 55% of oocyte donors prefer to stay in anonymity and 20% were not sure. In case that anonymity would be abolished, only 20% of the Czech oocyte donors would be willing to donate. There were no differences based on education (p = 0,358), age (p = 0,112), marital (p = 0,741) and maternal status (p = 0,691). (c) The main motivation to donation was altruism (helping infertile people), no less important factors were the finantial motivation and the testing own fertility. The altruistic motivation to donate was significantly related to the wish to remain anonymous (p = 0,0002). CONCLUSION: The typical Czech oocyte donor is young, mainly altruistic woman with her own child who wants to stay in anonymous relation to the couple whom she is helping as well as to the possible genetic offspring.

[Current trends in gamete donation - psychosocial and ethical issues]. / Soucasné trendy v psychosociální a etické problematice dárcovství gamet.

OBJECTIVE: To overview contemporary knowledge of legal and psychosocial rules in gamete donation. Previously, anonymous donation was preferred and recommended by experts but currently, with respect to the right to know the genetic origin of individuals, the relation to donor anonymity was reconsidered in many countries. There is a growing tendency to introduce the open identity system in gamete donation. Such system may guarantee that the child born after gamete donation may have receive the identification data of the donor of gametes. DESIGN: A review. SETTING: Clinic of Reproductive Medicine and Gynecology Zlin. METHODS: An overview of recent literature evaluating the influence of donor anonymity vs. open identity on psychosocial development of children born after gamete donation as well as on the quality of the relationship between parents and children in such families. CONCLUSION: New medical technologies usually overtake the developmental speed of ethics and psychology, and their impact on human society. Current trend to open identity is strong but there is no clear evidence that the open identity is of real importance for the healthy psychosocial development of a child born after gamete donation. Furthermore, there is no evidence that anonymity and secrecy of the gamete donation is harmful. In case of the consideration of the change in legal regulation in anonymity/open identity in gamete donation we would suggest the thorough consideration of all consequences.

[Influence of hCG glycosylation on its functions in female reproduction]. / Vliv glykosylace hCG na jeho funkce v zenské reprodukci.

OBJECTIVE: To review contemporary knowledge of the hCG molecule, its isoforms and the importance of glycosylation. Biologic variants and glycoforms of hCG have different biological activities and functions related to the control of menstrual cycle, conception, gestation as well as gynaecologic and non-gynaecologic malignancies. DESIGN: A review. SETTING: Department of Obstetrics and Gynaecology, University Hospital Olomouc. METHODS: To present own experience and an overview of recent literature in molecular biology, clinical biochemistry and clinical practice. CONCLUSION: Recent knowledge of the role of hCG glycosylation in physiologic and pathologic events in female organism will provide a better understanding of regulation of processes like ovulation (co-operation of pituitary hCG with LH), implantation and hemochorial placentation (invasivity of hyperglycosylated hCG). Some biologic variants and isoforms of hCG are important for the prediction of certain pathologies of pregnancy, prenatal screening of inborn errors (free beta hCG) as well as in the treatment of infertility.

[The influence of warmed infusion during C-section on the mother and child--a pilot randomised prospective study]. / Vliv ohrátých infuzních roztoku pri plánovaném císarském rezu na matku a plod - pilotní randomizovaná prospektivní studie.

AIM OF THE STUDY: To evaluate the influence of warmed infusions on the mothers and newborns well-being after C-section. DESIGN: Prospective randomized study. SETTING: Dept. of Anaesthesiology, Resuscitation and Intensive Care Medicine, Dept. of Obstetrics and Gynaecology, Faculty of Medicine and Dentistry Palacký University and University Hospital Olomouc. METHODS: Prospective randomized study including 29 pregnant women between 38th and 42nd week of pregnancy, that were indicated to elective caesarian section (SC) and from all of which the fetal distress was excluded. The women were randomized into four groups according to the temperature of applied infusion solution - colloids and afterwards crystalloids (pre-heated and non pre-heated), and according to the type of anaesthesia (general or SAB). These values were monitored in perioperative period: body temperature, differences of hemoglobin and hematocrit levels, tremor and pain level after the operation. In newborns the acid-base balance parameters and Apgar score were monitored. RESULTS: The study is still going on, however up to now some of the results suggest that usage of pre-heated infusion solutions is effective both for mother and child in increasing of their comfort in the perioperative period. In the group of actively heated mothers we have noticed: lower perioperative blood loss - differences in haemoglobin levels were 8 g/l compared to 15 g/l in the group of non heated mothers. The umbilical cord blood of the newborns whose mothers were not actively heated and were operated in subarachnoid anaesthesia had the lowest pO2 (0.9 kPa). The lowest level of umbilical cord blood lactate was noticed in the group of newborns whose mothers underwent the surgery under general anaesthesia and were actively heated. CONCLUSION: Up to now results suggest that the heating of mothers during the SC lowers the loss of body temperature, blood loss, and perception of pain by the mother. In combination with general anaesthesia it increases pO2 and lowers the levels of lactate in umbilical arteria in the newborns.

[Influence of the length of cultivation of no early cleavage embryos on the IVF success rate]. / Vliv délky kultivace embryí s pomalým nástupem delení na úspesnost IVF.

OBJECTIVE OF THE STUDY: To evaluate the success of treatment cycle after transfer of no early cleavage embryos in relation to the length of in vitro cultivation. TYPE OF THE STUDY: Retrospective study. SETTING: Dept. of Obstetrics and Gynecology, Faculty of Medicine and Dentistry, Palacky University and University Hospital Olomouc. METHODS: The IVF cycles with transfer of no early cleavage (NEC) embryos, ie. embryos with a delayed cleavage were evaluated in relation to the length of in vitro cultivation. The total of 338 embryos were cultured for three days (group 3D) and 204 embryos were cultured for 5 days (group 5D). The input parameters of the patients in these two groups were: their age, the percentage of fertilized oocytes and the number of transferred embryos. As the output parameters the number of cancelled cycles, clinical pregnancy rate (PR/ET), implantation rate (IR), and the number of pregnancy losses (AB) were evaluated. Finally, the results of these pregnancies were also evaluated. RESULTS: Both study groups had the same input parameters, except that fewer embryos were transferred in the group 5D than in 3D (1.8 ± 0.5 vs. 2.0 ± 0.5,p = 0.000). No cycles were cancelled in the group 3D while 33 cycles were cancelled in 5D (p = 0.000). The number of clinical pregnancies and implantation rate in the group 5D was significantly higher than in 3D (PR/ET 50% vs. 37%, IR 36% vs. 25%, p = 0.001). The groups did not statistically significantly differ in the number of pregnancy losses, or in the number of pregnancies that resulted in childbirth. CONCLUSION: In the 5D group with the longer in vitro cultivation statistically more pregnancies and implanted embryos were achieved, although the number of pregnancies that resulted in childbirth did not differ between groups.

[The occurence of genetic trombophilic markers in patients evaluated for infertility]. / Výskyt vybraných geneticky podmínených trombofilních markeru u pacientek podstupujících lécbu neplodnosti.

OBJECTIVE: To assess and compare the frequency of selected gene mutations of thrombophilic markers (FV Leiden, FII prothrombin G20210A and MTHFR C677T) in patients with primary and secondary infertility. DESIGN: Retrospective study. SETTING: Institute of normal anatomy, Faculty of Medicine and Dentistry, Palacky University Olomouc. METHODS: The study included 92 patients with primary infertility and 89 patients with secondary infertility. Indications for examination of these mutations were following: a positive family or personal history, a positive obstetrical history or a repeated failure of assisted reproduction treatment. RESULTS: According to our anticipation, women with the secondary infertility were significantly older(p < 0.0005) than those with primary infertility. No mutations of genes of examined thrombophilic markers (FV, FII and MTHFR), either alone or in combination, were found in only 8.7 % patients with primary infertility and in 5.6 % patients with secondary infertility. Significantly higher frequency of factor Leiden(p < 0.02) was observed in women with secondary infertility. There were no significant differences in the frequency of detected mutations of the remaining factors. CONCLUSION: Based on our findings we suggest that the assessment of selected gene mutations of thrombophilic markers should be a part of the diagnostic algorithm in patients with positive history for thrombophilic disorders.

Increased lipid peroxidation and abnormal fatty acid profiles in seminal and blood plasma of normozoospermic males from infertile couples.

BACKGROUND: Reactive oxygen species (ROS) are essential for sperm function. However, excessive ROS production can impair sperm function and might be a factor contributing to male infertility. METHODS: We investigated the levels of arachidonic acid (AA) and docosahexaenoic acid (DHA) as well as lipid peroxidation, as represented by thiobarbituric acid reactive species (TBARS), in blood and seminal plasma of 38 normozoospermic males from infertile couples (NSI-males), compared with that of 17 fertile volunteers (FV-males). RESULTS: TBARS levels in blood and seminal plasma were higher in NSI-males than in FV-males (P < 0.0002, P < 0.0003, respectively), as were AA levels (P < 0.0003, P < 0.00004, respectively). On the contrary, the blood and seminal plasma levels of DHA were lower in NSI-males than in FV-males (P < 0.02 and P < 0.05, respectively). The AA/DHA ratios in blood and seminal plasma were higher in NSI-males than in FV-males (P < 0.003, P < 0.0007, respectively). Significant correlations between seminal and blood plasma levels of TBARS (P < 0.0001, r = 0.548), AA (P < 0.0001, r = 0.571) and DHA (P < 0.0001, r = 0.506) were found. CONCLUSIONS: Our data provide new insight into lipid metabolism in male infertility and indicate that systemic oxidative stress resulting in increased lipid peroxidation and an altered fatty acid profile may be, at least in part, responsible for infertility even in normozoospermic males.

[Comparison of reactive oxygen species production in neat semen and washed spermatozoa]. / Porovnání produkce reaktivních kyslíkových cástic v nativním ejakulátu a v suspenzi spermií.

OBJECTIVE: To determine Reactive Oxygen Species (ROS) production in neat semen and spermatozoa suspension using chemiluminescence and to examine correlation between both methods. SUBJECT: Prospective laboratory study. SETTING: Department of Obstetric and Gynecology, University Hospital, Olomouc. METHODS: The study included fertile volunteers (FV, n = 17), men from infertile couples (NM, n = 19) and men with idiopathic infertility (NMI, n = 15). ROS levels were determined by the same method in neat and washed semen samples. RESULTS: The ROS production in neat semen was lower than that in spermatozoa suspension. There was no significant diference in ROS production between volunteers and males from infertile couples. There was a significant correlation between log ROS in neat semen and in spermatozoa suspension in studied groups (FV r = 0.85, p = 1.5 x 10(-5); NM r = 0.76, p < 2 x 10(-4); NMI r = 0.75, p < 1.5 x 10(-3)). CONCLUSIONS: Measurement of ROS in neat semen is simpler, faster and better reflecting the actual level of oxidative stress than the same measurement in spermatozoa suspension. The implementation of this method can complement the algorithm of diagnostics and treatment of male infertility and be helpful in selection of patients for antioxidant or antibiotic treatment.

[Oxidative stress in diagnosis and treatment of idiopathic male infertility]. / Oxidacní stres v diagnostice a lécbe idiopatické muzské neplodnosti.

OBJECTIVE: To summarize available data concerning reactive oxygen species in sperm physiology and male infertility. DESIGN: Review article. SETTING: Dept. of Obstetrics and Gynecology, Faculty of Medicine and Dentistry, Palacky University and University Hospital Olomouc. METHODS: Compilation of published data from scientific literature. CONCLUSIONS: The article describes the importance of reactive oxygen species and their role in male reproductive health.

[EmbryoGlue the transfer medium with hyaluronan in the IVF+ET program]. / Transferové médium s hyaluronanem embryoGlue v programu IVF+ET.

OBJECTIVE: To assess, if the replacement of serum albumin by recombinant human albumin and hyaluronan in EmbryoGlue transfer medium effects embryo implantation within IVF+ET programme. DESIGN: A prospective study. SETTING: Department of Obstetric and Gynecology, University Hospital, Olomouc. METHODS: 297 IVF cycles with the embryotransfer (ET) were evaluated. The patients were divided into 2 groups according to the type of transfer medium (culture medium; G2.2, n = 112 and the transfer medium EmbyoGlue (EG) n = 185) and to three subgroups according to the age of the patients (< 30, 30-38, > 38). In both groups the following entry parameters were compared: average age, number of oocytes obtained, fertilization rate (FR), number of frozen embryos, number of transferred embryos. The outcome parameters compared were: pregnancy rate (PR), implantation rate (IR), abortions (AB) and the number of multiple pregnancies. RESULTS: In the group of patients up to 30 (EG = 72 vs. G2.2 = 51) and over 38 years of age EG = 18 vs. G2.2 = 18) no differences in input and output parameters have been discovered, while in the group of patients between 30 and 38 years of age (EG = 94 vs. G2.2 = 44) the results of EG group embryo implantation have been improved significantly (IR 27.7% vs. 153%, p < 0.01). No differences in any other output parameters have been encountered (PR 43.6% vs. 39.5%, AB 7.3% vs. 11.7%). CONCLUSION: Human serum albumin can be replaced by hyaluronan as a sole macromolecule in a human embryo transfer medium and by recombinant human albumin, without reduction of pregnancy rate (PR) and implantation rate (IR). Not only similar results were achieved in EG medium compared to currently used G2.2 medium, but in the group of 30-38 year old patients a statistically significant positive influence of EG on the implantation of embryos has been observed.

[Lycopene therapy in male infertility]. / Lykopen v terapii muzské neplodnosti.

OBJECTIVE: To summarize available data concerning lycopene and male infertility treatment. DESIGN: Review article. SETTING: Dept. of Obstetrics and Gynecology, University Hospital Olomouc. METHODS: Compilation of published data from scientific literature. CONCLUSIONS: The article describes occurrence, biochemistry, metabolism of lycopene and its role in male reproductive health.

Endometrial expression of the estrogen-sensitive genes MMP-26 and TIMP-4 is altered by a substitution protocol without down-regulation in IVF patients.

BACKGROUND: The aim of this study was to analyse the effects of an estradiol (E(2))-progesterone substitution protocol on the endometrial expression of estrogen-sensitive genes during the peri-implantation period. METHODS: Peripheral blood and endometrial biopsies were obtained from 13 infertile women both in a natural cycle (NC), on days 5 and 7 after ovulation (NC5, NC7), and in an artificial (substituted) cycle (AC), on days 5 and 7 of progesterone addition (AC5, AC7). Estrogen receptor-alpha (ERalpha) and progesterone receptor (PR) were assayed by immunohistochemistry. Matrix metalloproteinase-26 (MMP-26) mRNA and tissue inhibitor of metalloproteinase-4 (TIMP-4) mRNA were semiquantitatively assessed in tissue sections using in situ hybridization (ISH) and quantified in tissue extracts using real-time PCR. RESULTS: Levels of both E(2) and progesterone were higher in the peripheral blood in AC than in NC. Also on day AC5, expressions of ERalpha, PR and MMP-26 mRNA (focally) were increased in the epithelium and TIMP-4 mRNA in the stroma. Expression levels of these genes dropped significantly between AC5 and AC7, but not between NC5 and NC7. Abnormally high levels in AC5 samples suggest overstimulation with E(2), and the rapid decrease between AC5 and AC7 suggests overstimulation with progesterone. CONCLUSIONS: In ACs, increased levels of E(2) in the blood exaggerate the endometrial expression of estrogen-sensitive genes, whereas higher levels of progesterone in the blood in the secretory phase exaggerate the drop in expression of these genes. Dramatic variations in the gene expression may not be optimal for the implantation process.

[Embryo quality evaluation according to the speed of the first cleavage after IntraCytoplasmic Sperm Injection (ICSI)]. / Kvalita embryí po ICSI v závislosti na rychlosti nástupu prvního bunecného delení.

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.

[Embryo quality evaluation according to the speed of the first cleavage after conventional IVF]. / Kvalita embryí po klasickém IVF v závislosti na rychlosti nástupu prvního bunecného delení.

OBJECTIVE: To evaluate morphological parametres of embryos obtained in the process of conventional IVF. DESIGN: A prospective study. SETTING: Centre of Assisted Reproduction, Dept. of Obstetrics and Gynecology, Palacký University Medical School, Olomouc. METHODS: In the present study 549 embryos developing after conventional IVF in the period of 2001-2004 were evaluated. The beginning of the mitotic cleavage was assessed within the interval of 24 - 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 - with regular blastomeres, without fragmentation, B - with irregular blastomeres or with fragmentation below 30%, C - with fragmentation 30-50%, D - with fragmentation above 50%. The speed of further cleavage and average number of blastomeres were evaluated on days two and three of cultivation. Statistical analysis was preformed in the Palacký University Computer Centre. The chi-square test and t test for independent samples were used. RESULTS: EC embryos were found in 45.5%, BDPN in 33.5% and PN in 21.0%. 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 comparisson 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 difference was found also between the EC and BDPN groups (p = 0.000). CONCLUSION: The speed of the first cell cleavage is a useful additional criterion for the embryo selection for ET. EC embryos usually have better morphology and more blastomeres that the BPDN and PN ones.

[Prevention of multiple pregnancy after IVF/ICSI by elective single embryo transfer--pilot study]. / Prevence vícecetného tehotenství po metodé IVF/ICSI elektivním transferem jednoho embrya--pilotní studie.

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.

[The analysis of preliminary spermatological parameters in potential donors of semen]. / Analýza spermiologických parametru vstupního vysetiení potenciálních dáircu spermí.

OBJECTIVE: To contribute to the contemporary discussion on the male factor infertility through the evaluation of the basic spermatological parameters in men recruiting for semen donation. DESIGN: A retrospective demographic study. SETTING: Dept. of Obstetrics and Gynaecology, Dept. of Biology, Faculty of Medicine, Palacký University, Olomouc. METHODS: A total of 108 men aged 18 to 35 years came to the preliminary examination for the potential semen donors between January 2000 and April 2004. Spermatological analysis of the fresh ejaculates obtained by masturbation was performed according to WHO guidelines. The following parameters were evaluated: the volume of ejaculate, sperm concentration and total percent motility. RESULTS: Normospermia was found in only 46 men (42.6%). Three males (2.8%) were azoospermic. The most frequent problem was asthenospermia, which was documented in 49 (46.6%) men. CONCLUSION: The analysis of the obtained data may suggest a marked semen quality decline in the Czech male population. In our opinion, further multicentric epidemiological studies concerning fertility potential in Czech male population seem highly needed.

[Protein bcl-2 expression in the secretory endometrium after oral estrogen-progesterone replacement]. / Exprese proteinu bcl-2 v sekrecním endometriu po perorální estrogen-progesteronové substituci.

OBJECTIVE: To compare the changes of the protein bcl-2 expression, regulatory mechanism in the process of apoptosis, in the secretory endometrium throughout natural and artificial (estrogen-progesterone substitution) cycles in the same group of patients. DESIGN: A prospective study. SETTING: Dept. of Obstetrics and Gynecology, University Hospital Olomouc, Dept. of Histology and Embryology, Faculty of Medicine, Palacký University in Olomouc, Dept. of Obstetrics and Gynecology, Bata Hospital, Zlín. METHODS: Endometrial samples (n=56) were obtained on days 5 and 7 after ovulation, progesterone addition, resp. Patients (n=14) included in the study had regular menstrual cycle, age under 40, BMI range 20-32 and basal FSH level range 5-9 IU/l. The collected samples were processed routinely and bcl-2 was estimated by indirect three level imunohistochemic method. RESULTS: We demonstrated the bcl-2 expression in all evaluated layers (epithelial surface, stroma, glands) of the mid-secretory endometrium in both cycles. No difference was found in bcl-2 expression between days 5 and 7 in the secretory phase of the natural cycle. In artificial cycles higher bcl-2 expression was found only in epithelial surface (p<0.05) between days 5 and 7. On day 7 higher bcl-2 expression was found in the artificial cycle in the endometrial epithelial surface (p<0.001) as well as in spongy layer (p<0.01) and compact layer (p<0.05) comparing to day 7 of the natural cycle. CONCLUSION: The bcl-2 expression in the mid-secretory endometrium is significantly higher in the cycle with estrogen-progesterone substitution comparing to the natural cycle. These changes were more significant in endometrial and glandular epithelium than in stromal cells.

[Efficiency of cryoembryotransfer using embryos frozen in the pronuclear stage]. / Uspesnost cyklu s prenosem embryí zamrazených ve stadiu prvojader.

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.