ASXL1 gene alterations in patients with isolated 20q deletion.

Deletion 20q is a recurrent abnormality in myeloid malignancies. In our previous study, we identified fusion of the additional sex combs-like 1 (ASXL1) and teashirt zinc finger homeobox 2 genes in a patient with myelodysplastic syndrome. The objective of this study was to determine the frequency of ASXL1 breakpoints in a cohort of 36 patients with deletion 20q as the sole cytogenetic aberration. A combination of molecular cytogenetic methods was used to confirm ASXL1 gene alterations in 19 of the 36 patients, and the determination of ASXL1 gene changes in patients with deletion 20q revealed clinical and prognostic impacts.

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.

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.

[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.

The assessment of array comparative genomic hybridization in complex karyotype analyses.

Molecular-cytogenetic methods were used to analyse and specify complex genome rearrangements in malignant cells. Twelve samples of bone marrow cells were collected from patients with myelodysplastic syndromes (MDS). The complex karyotypes were examined by multicolour fluorescence in situ hybridization (mFISH), high-resolution multicolour banding (mBAND) and array comparative genomic hybridization (aCGH). For aCGH, DNA was isolated from fixed bone marrow cells in methanol and acetic acid and amplified by whole-genome amplification. Three samples were analysed by the oligonucleotide array NimbleGen on the basis of full service. BAC-based Haematochips (BlueGnome) were used for the other nine samples. Sensitivity and detection limits of both methods were compared. The results obtained by mFISH/mBAND were in most cases confirmed by the microarray technique. aCGH detected 43 unbalanced chromosomal changes that were also identified by classical cytogenetics and FISH. Moreover, aCGH discovered 14 additional changes. Cryptic amplifications and deletions were characterized with a resolution of 0.5 Mb. In one bone marrow sample with suspected monosomy 5 detected by conventional cytogenetic analysis, aCGH revealed a 22.3 Mb region of chromosome 5 inserted in another autosome within the complex karyotype. Amplified DNA was successfully used for aCGH in 11 out of 12 cases, improving resolution of unbalanced chromosomal aberrations. The combination of both approaches brought more detailed description of complex karyotypes and is highly recommended.

The role of telomeres and telomerase complex in haematological neoplasia: the length of telomeres as a marker of carcinogenesis and prognosis of disease.

Human telomeres (discovery of telomere structure and function has been recently awarded The Nobel Prize) consist of approximately 5-12 kb of tandem repeated sequences (TTAGGG)n and associated proteins capping chromosome ends which prevent degradation, loss of genetic information, end-to-end fusion, senescence and apoptosis. Due to the end-replication problem, telomere repeats are lost with each cell division, eventually leading to genetic instability and cellular senescence when telomeres become critically short. Stabilization of the telomeric DNA through telomerase activation, unique reverse transcriptase, or activation of the alternative mechanism of telomere maintenance is essential if the cells are to survive and proliferate indefinitely. Telomerase is expressed during early development and remains fully active in specific germline cells, but is undetectable in most normal somatic cells. High level of telomerase activity is detected in almost 90% of human tumours and immortalized cell lines. The hematopoietic compartment may develop genetic instability as a consequence of telomere erosion, resulting in aplastic anaemia (AA) and increased risk of myelodysplastic syndrome (MDS) and acute myeloid leukaemia (AML). Genetic instability associated with telomere dysfunction (i.e. short telomeres) is an early event in carcinogenesis. The molecular cytogenetic method telomere/centromere fluorescence in situ hybridization (T/C-FISH) can be used to characterize the telomere length of hematopoietic cells. This review describes recent advances in the molecular characterization of telomere system, the regulation of telomerase activity in cancer pathogenesis and shows that the telomeric length could be a potential clinical marker of hematologic neoplasia and prognosis of disease.

Dasatinib in imatinib-resistant or -intolerant CML patients: data from the clinical practice of 6 hematological centers in the Czech Republic.

Dasatinib is effective second line treatment for patients with chronic myeloid leukemia (CML) resistant or intolerant to imatinib. We report here the first experiences with dasatinib therapy in 71 CML patients resistant or intolerant to imatinib from the real clinical practice of 6 hematological centers in the Czech Republic. Dose 100 mg daily and 70 mg twice daily was administered to patients with chronic phase (CP) and advanced phases (AP) CML. In chronic phase (n=46), complete hematological reponse (CHR) was achieved in 97%, major cytogenetic reponse (MCgR) in 77% and complete cytogenetic response (CCgR) in 67%. Major molecular reponse (MMR) was achieved in 19/31 patients in median of 10 months. In advanced phase (n=25), CHR was attained in 77%, MCgR in 39%, CCgR in 33% and MMR in 2/18 patients. Eleven different baseline mutations were followed up in 15 patients. Dasatinib eliminated mutations in most of the patients, but 3 patients acquired a new one. Novel mutations were detected under dasatinib therapy in 2 patients. Dasatinib was well tolerated, cytopenias were common and was managed by dose modification. The estimated progression free survival (PFS) at 12 months was 97+/-3% in CP and 62+/-21% in AP. The median time to treatment failure was 605 days in AP while it was not reached in CP patients. Our clinical experiences, described here, confirmed that dasatinib is associated with high response rates especially in imatinib resistant or intolerant CML patients in chronic phase.

Telomere length, molecular cytogenetic findings, and immunophenotypic features in previously untreated patients with B-chronic lymphocytic leukemia.

UNLABELLED: Telomere length was evaluated by terminal repeat fragment method in 66 previously untreated patients with B-chronic lymphocytic leukemia (B-CLL) to ascertain whether telomere shortening was associated with genomic aberrations, immunoglobulin variable heavy chain (IgVH) mutational status, CD38 and ZAP-70 expression, and telomerase activity. Chromosomal aberrations were present in peripheral blood cells of 73% patients (48/66), no difference in telomere length between patients with good and intermediate prognosis according to cytogenetics was found. Association between telomere length and IgVH mutational status, ZAP-70 and CD38 expression was proved as significantly shorter telomeres in patients with unmutated IgVH status (p=0.01) and ZAP-70 positivity (p=0.01) and CD38 positivity (p=0.05) were detected. Telomerase activity was positive in 11 patients out of 21 examined, correlation between telomere length and telomerase activity was found (p=0.05). Telomere length and telomerase activity in combination with other prognostic parameters complete the risk profile of B-CLL patients and might serve for an easy decision on optimal treatment strategy. KEYWORDS: B-chronic lymphocytic leukemia, telomere length, telomerase activity, chromosomal aberrations, prognosis.

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.

Clonal evolution in chronic lymphocytic leukemia studied by interphase fluorescence in-situ hybridization.

The results of repeated interphase fluorescence in-situ hybridization (I-FISH, FISH) examination of 97 CLL patients and correlation of these findings with IgVH hypermutation status, ZAP-70 and CD38 expression are presented. The appearance of new, FISH-detectable, genomic aberrations during disease course, described as clonal evolution (CE), was observed in 26% of patients. The most frequent newly acquired cytogenetic abnormality was 13q deletion in 64% (16/25). In contrast to earlier studies, there was no correlation found between CE and either one of single negative prognostic factors (unmutated IgVH; CD38 positivity; ZAP-70 positivity). However, the combination of all three negative factors correlated with CE highly significantly (p=0.005) and moreover, also with a shift from lower to higher FISH risk category (p=0.010). As the prognostic data were known in all patients, this study represents the complete insight on the association of CE and other risk parameters in CLL.

[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.

Combined molecular biological and molecular cytogenetic analysis of genomic changes in 146 patients with B-cell chronic lymphocytic leukemia.

Interphase fluorescence in situ hybridization was used to detect common deletions in B-CLL patients as well as trisomy 12 and aberrations of IgH gene complex at 14q32.33 where we evaluated not only translocation-like signal pattern but also deletions. 120 (82%) patients showed genetic changes - del(13)(q14) 95 (62%), deletion of ATM gene 22 (15%), deletion of p53 gene 25 (17%) and trisomy 12 was proved in 18 (12%) cases. IgH rearrangements were detected in 45 (31%), split of the signals in 11 (8%), deletion of 3' segment flanking IgH gene in 5 (3%) and deletions of variable segment in 29 (20%) patients. Although deletions of 3' segment flanking IgH gene complex are supposed to have an adverse prognostic impact and the genetic background of variable segment deletions is believed to be most probably physiological, we assumed a detailed mapping of the 14q32.33 region will be needed to unravel these mysteries.

[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.

[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.

Prognostic relevance of the FAB morphological criteria in chronic lymphocytic leukemia: correlations with IgVH gene mutational status and other prognostic markers.

Morphological examination is the routine first step in the diagnosis of hematological malignancies, including chronic lymphocytic leukemia (CLL). Atypical cell morphology according to the FAB criteria is known to herald disease progression. Several years ago, it was proposed that FAB morphology at diagnosis had a considerable prognostic impact. However, this proposal has not been widely adopted in practice. Thus we questioned the prognostic value of the morphological examination, which was performed retrospectively in 88 patients out of our 110 institutional registry patients (70 males and 40 females, median age 57 yrs) with CLL at diagnosis. We related the results to the more modern prognostic markers. Atypical FAB morphology was shown to correlate with IgVH gene mutation status, trisomy of chromosome 12 and deletion of 17p detected either by conventional G-banding or by fluorescence in situ hybridization (FISH) analysis. The correlation of FAB morphology with CD38 antigen expression or with the histopathological pattern of bone marrow infiltration was not significant. Overall survival (OS) data were available for 84 morphologically examined patients. The patients with atypical morphology (64 patients) had a significantly shorter OS (103 months) than the 20 patients presenting with typical CLL morphology (237 months; p=0.03). Only the mutation status of IgVH genes correlated more closely with OS (p=0.002). Of note, there was no leukemia-related death within "unmutated" cases who had typical FAB morphology (p=0.14), and vice versa, the mutation status had a significant prognostic impact within the morphologically atypical cases (p=0.01). Thus FAB morphology and the mutation status may yield complementary prognostic information. OS was affected both by the presence of cytogenetic aberrations (p=0.03) - most adversely by deletions of 17p and 11q, and by CD38 expression (p=0.003). We conclude that careful examination of peripheral blood smears according to FAB is a simple, cheap and valuable tool in the first-line assessment of prognosis of CLL patients and should not be overlooked even in 3rd millennium when more sophisticated prognostic markers are at hand. This ought to be confirmed in larger prospective studies with multivariate analysis of data.

[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.

[Factors influencing success rate of FISH in pre-implantation genetic diagnostics]. / Faktory ovlivnující úspesnost FISH pri preimplantacní genetické diagnostice.

OBJECTIVE: Information on preimplantation diagnostics (PGD) by means of the method of fluorescent in situ hybridization (FISH) and some factors, which principally influence the results obtained with this technique from the point of view of a molecular cytogeneticist. Critical steps of work with one cell in PGD-FISH are summarized based on data from literature as well as from the authors, own experimental experience. The individual steps are analyzed and possibilities of execution are presented. TYPE OF STUDY: A review. SETTING: Institute of Medical Genetics and Fetal Medicine of Faculty Hospital and Medical Faculty of Palacky University, Center of Assisted Reproduction at Obstetric-Gynecological Clinic, Faculty Hospital, Olomouc. SUBJECT AND METHODS OF STUDY: Introduction into the PGD problem. A survey and analysis of factors decisive for the success of the FISH method for PGD. - Selection of embrya for the experimental phase of PGD-FISH. - Selection of fixation method and pretreatment, application of suitable probes. - Localization of the fixed cell. - Interpretation of the results obtained. Recommendation of the mode of their successful solution. CONCLUSION: The paper presents a survey of the PGD-FISH method. Some critical aspects in the analysis of unicellular preparation are pointed out and evaluated and possibilities and procedures of the optimal solution are suggested.