[Application of SNP array method in prenatal diagnosis]. / Aplikace metody SNP array v prenatální diagnostice.

OBJECTIVES: SNP array (array method using Single Nucleotide Polymorphisms) enables to detect cytogenetically undetectable submicroscopic alterations (microdeletions, microduplications), which could be also causative for ultrasonographic anomalies of fetus. This article describes the principle, advantages, disadvantages and application possibilities of the SNP array method in prenatal diagnosis. The ten month experience with SNP array use in prenatal diagnosis is presented. DESIGN: Prospective study. SETTINGS: Gennet, Prague. MATERIAL AND METHODS: During the period from April 2010 to January 2011 we performed 110 SNP array analyses of fetal DNA: 14 chorionic villi samples (CVS), 88 amniotic fluid samples (AMC), 1 cord blood sample and 7 miscarriage samples. Laboratory tests were carried out on DNA from both cultured and uncultured fetal cells. Examinations were performed in fetuses with sonographic abnormal findings having normal karyotype. In addition 14 fetal cytogenetic abnormalities were solved. SNP array analysis was performed using Illumina InfiniumHD HumanCytoSNP-12 chip. All data were analysed by Illumina KaryoStudio and GenomeStudio software. RESULTS: SNP array analysis was performed in 108 fetuses (only 2 examination failures, 1.8%). In total, we detected CNV (copy number variation) in 29 samples (29/108 = 27%). 15% (16/108) of fetuses with abnormal ultrasound findings were found to carry clinically relevant CNV. Probably benign CNVs were found in 8 samples (8/108 = 7%) and in additional 5 CNVs parental samples have not been analysed yet. Excluding karyotypically abnormal cases clinically relevant CNVs were found in 10% of fetuses (9/94). In all cases with de novo chromosomal aberration the clinical relevancy was clarified (imbalances in 50%). CONCLUSION: Our data suggest that SNP array analysis is a relevant and useful technique in prenatal diagnosis.

[Abortion induction in IInd trimester]. / Indukce potratu ve II. trimestru na Gynekologicko porodnické klinice FN na Bulovce.

STUDY AIM: Analysis of the set of women that underwent the termination of pregnancy in IInd trimester at the OBGYN clinic, Teaching Hospital Na Bulovce at interval of years 2006-2007. We appreciated the effect of method and compared with references in the literature. TYPE STUDY: Retrospective descriptive study. SEATTING: OBGYN clinic of the 1st faculty of Charles University, teaching hospital Na Bulovce, Prague. METHOD: Retrospective analysis of the set of women that underwent termination of pregnancy in the IInd trimester. Data are obtained from medical documentation and statistically processed. The analysis treats with 50 cases, in all of them, cervix was prepared with hydrophile dilators. In 37 cases were subsequently handed up prostaglandins intraamnially, in two cases generally intravenously, in two cases vaginally and 9 pregnancies were finished directly by dilatation and curettage without endeavour about expulsion. RESULTS: From 37 women after intraamnial administration of prostaglandins, 35 (94.6%) aborted successfully. Average time from amniocentesis to expulsion was 13 hours 45 minutes, 23 women aborted to 24 hours (62.2%). Undesirable effects were present in 12 cases (32.4%). CONCLUSION: Our record of local intraamnial administration of prostaglandins appears to be effective method. On the other hand, other methods are described in recent literature, which appear to be more efficient and have smaller occurrence of adverse effects.

[Development of the pregnancy and labor in patient with systemic lupus erythematosus]. / Prubeh tehotenství a porodu u pacientky se systémovým lupus erythematodes.

OBJECTIVE: Pregnancy and its outcome in a woman with systemic lupus erythematosus. SUBJECT: Case report. SETTING: Obstetrics and Gynecology Department, Charles University, 2nd Medical Faculty and Faculty Hospital Motol, Prague, Cardiocentrum, University Hospital Motol, Prague. SUBJECT: Patient with systemic lupus erythematosus (SLE) was admitted to our department at 31 weeks of gestation with diagnosed fetal bradycardia. Detailed prenatal echocardiography revealed complete atrioventricular block (AVB III) with atrial rate of 140 b.p.m. and ventricular rate of 70 b.p.m., fetoplacental failure was not present at initial examination. Patient was hospitalised for the remainder of the pregnancy, which continued to full-term without complications. During delivery, fetal pulse oxymetry was utilized to monitor fetal state saturation levels. Labour proceeded without complications with an Apgar score of 9-9-9. The newborn was observed at the ICU and the Cardiac center and deemed to be in stable condition with AVB III without any signs of heart failure and without the necessity for pacemaker implantation. CONCLUSION: In cases of diagnosed fetal bradycardia, it is beneficial to expand the methods of fetal monitoring during labour. Pre-, peri-, and postnatal observation of pregnant women diagnosed with SLE and fetal bradycardia should be done in specialised facilities that are able to provide the appropriate level of care for mother and infant.

[Diagnosis and therapy of gestational diabetes mellitus]. / Diagnostika a terapie gestacního diabetes mellitus.

In 1999-2000 at the Medical Clinic Motol Faculty Hospital in collaboration with the Gynaecological Clinic of the Hospital a group of 55 pregnant women with the diagnosis of gestational diabetes (GDM) were followed up. The objective of the investigation was to find out how in the investigated area (detachment area of the Gynaecological and Obstetric Clinic Motol Faculty Hospital) GDM is diagnosed at present, how it is treated and what is the percentage of perinatal morbidity in the investigated group. The mean age of the investigated women was 32.3 +/- 4.5 years. The presence of risk factors for the development of GDM was found in 59.8% of the examined women. 65.7% women had a positive gynaecological case-history. GDM was detected most frequently during the 30th week of pregnancy, in 25% women in the 35th and later week of gestation. In 52% the diagnosis of GDM was established only on hospital admission on account of complications of pregnancy. The mean HbA1C level during detection of gestational diabetes was 6.81 +/- 0.41%. The majority of women -91.1%--were treated by diet, 8.9% women had insulin treatment. The prevalence of diabetic foctopathy was 48.3%. The mean weight of the offspring of diabetic mothers was 3350 g +/- 248 g, the mean length was 49.6 +/- 6.3 cm. No stillbirth was recorded. One infant suffered from an inborn developmental defect (morbus Down). The results provide evidence not only of late diagnosis of GDM (after the 28th week of gestation) but also of inadequate screening in the field, as GDM is frequently detected only during complications of pregnancy.

[Medical genetics in reproductive medicine]. / Vyuzití lékarské genetiky v reprodukcní medicíne.

Reproductive genetics (RG) is another new field of medical genetics, integrated with reproductive medicine, assisted reproduction and developmental genetic. RG is closely linked to the perioconceptional prevention, perinatology, ultrasound and biochemical screening in the end of the first and beginning of the second trimesters. RG is based on the system of specialized genetic counseling, clinical cytogenetics, molecular cytogenetics and molecular genetics to provide prefertilization, preimplantation and classical prenatal diagnosis in the Ist to IIIrd trimesters. Thus, RG is part of the fetal medicine and therapy. The six years experience with RG is summarized. A system of the specialized health care, organized, if possible in one integrated center of RG and reproductive medicine (RM) is presented. Reproductive medicine provides all necessary clinical gynecological and andrological surveillance, with assisted reproduction and further obstetrical ultrasound examinations, including nuchal translucency measurements and 2D, 3D ultrasound, echocardiography examinations, if indicated, as well as the invasive method of prenatal diagnosis and perinatology care. Specialized genetic counseling and cytogenetic analysis, if indicated, should be offered to all partners with reproductive disorders as well as to oocyte donors. Chromosome anomalies are disclosed in 6% of men with abnormal sperm analysis as well as in women with severe reproductive disorders. In males with severe oligo, azoospermia, the sperm aneuploidy analysis by molecular cytogenetic methods is recommended. Advised is also the molecular genetic detection of Y chromosome microdeletions, which is detected in 9% of our azoospermic men with deletions in AZFb region. CFTR gene mutations and intron 8 and 10 polymorphism examination is provided not only in men with obstructive azoospermia (CBAVD), but also if severe oligospermy with less than 1 x 10(6) sperm/ml is detected. Molecular genetic analysis of thrombophilic mutations of factor II., V. (Leiden) and MTHFR gene in unexplained recurrent abortions and in cases with unsuccessful IVF is part of the diagnostic strategy. The population frequencies of carriers of mutations of factor II. (2.3%), factor V.-Leiden (5.7%) and MTHFR gene (38%) were determined. The laser biopsy of the first polar body and of blastomeres was introduced for FISH analysis of chromosome aneuploidies. Quantitative fluorescent PCR (QFPCR) detection is used for testing of the most frequent delta F508 CFTR gene mutation and the most frequent aneuploidies of chromosome 13, 18, 21, X and Y. QFPCR was successfully tested for male fetal sex examination from partially purified fetal cells in the maternal blood. The first trimester ultrasound and biochemical screening is recommended to all successful pregnancies after different IVF methods. If borderline levels of first trimester biochemical screening of PAPP-A protein and beta hCG are detected without pathological ultrasound findings, classical triple test of biochemical screening in 16th week of gestation is recommended. If pathological results of ultrasound and biochemical screening are disclosed, invasive prenatal genetic diagnosis is indicated as well as in pregnancies after ICSL, if there is not any obstetrical contraindication.

[Intraumbilical therapy of fetal supraventricular tachycardia]. / Intraumbilikální lécba supraventrikulární tachykardie plodu.

The supraventricular tachycardia represents the life threatening disease, which may cause severe heart failure or even during foetal life. The authors present case report of the foetus aged 23 weeks of gestation in whom the supraventricular tachycardia was resistant to standard transplacental treatment by using digoxin and sotalol. The successful rhythm conversion was achieved by intracordal infusion of amiodarone. Further uncomplicated course of pregnancy reached term and healthy boy was subsequently born without having additional psychomotoric complications.

[Regression of hCG in various types of molar pregnancies--clinical course and prognosis]. / Regrese hCG u ruzných typu molárních tehotenství--klinický prubeh a prognóza.

OBJECTIVE: To evaluate spontaneous regression curves of hCG serum positivity in patients with surgically treated molar pregnancies. Comparison of complete, partial and invasive mole. The study should result in optimalisation of follow up criteria of molar pregnancies in respect to their potential malignant change. DESIGN: Retrospective comparative clinical study. SETTING: Obst. Gyn. Dpt., Oncogynecology div., 2nd Medical Faculty, FNM, Charles University Prague, Pathology Dpt., 2nd Medical Faculty, Institute of Biology and Medical Genetics. METHODS: Evaluation of spontaneous regression curves of serum hCG levels in 104 molar pregnancies. 46 patients with partial hydatiform mole, 48 patients with complete hydatiform mole, 10 patients with invasive mole. Serum hCG levels were detected by radioimunoassay (RIA) in the first period and imunochemoluminisent assay (LIA) in the second period. Regression curves of hCG positivity in particular moles were statistically evaluated by Fischer test and t-test. RESULTS: There is statistically significant difference in spontaneous regression of hCG positivity in different types of molar pregnancies. Recommended criteria for gestational trofoblastic disease (GTD) diagnosis and follow up are fully applicable in clinical practice. There is exception in partial hydatiform moles, where plateau in hCG regression does not necessarily implicate chemotherapy in patient with good compliance. CONCLUSION: Early diagnosis of GTD predominantly due to the widespread use of ultrasonography changes classical clinical features of molar pregnancies. Spontaneous regression in hCG positivity in serum is more rapid in patients with partial hydatiform mole, slower in complete hydatiform mole and invasive mole. There is no significant change in malignant potential regarding early detection and treatment.

[Sex behavior of women seeking hormonal contraception and women seeking induced abortion]. / Sexuální chování zadatelek o hormonální kontracepci a zadatelek o umelé prerusení tehotenství.

Using an anonymous questionnaire, the authors assessed the sexual and contraceptive behaviour of 130 applicants for induced abortions and 330 applicants for hormonal contraception. Between the two groups these main differences were found: applicants for abortion used less frequently effective contraception and used more frequently unreliable contraceptive methods. This difference was obvious during the first intercourse as well as during later life. The use of contraception in this group is more limited also as compared with the nationwide average. In the case-history of applicants for abortion previous abortions were more frequent, as well as prostitute behaviour and the prevalence of venereal diseases. As regards heterosexual behaviour the applicants for abortion and applicants for hormonal contraception did not differ substantially. According to the authors the differences between the examined groups depend more on personality traits than sexuological characteristics.

[Results and complications of intrauterine transfusion in fetuses with and without hydrops]. / Výsledky a komplikace intrauterinní transfuze u hydropických a nonhydropických plodu.

The group comprises 50 intrauterine transfusions in 15 patients with a severe form of foetal erythroblastosis. The authors describe the results and complications in hydropic and non-hydropic foetuses, diagnosed by ultrasound before transfusion treatment was started. In the group of non-hydropic foetuses the success rate of transfusion treatment was 90.1% and 10 neonates were born with a mean weight of 2030 g. The mean gestation period was 34 weeks. From four hydropic foetuses it proved possible to save only one, the success rate in this group was only 25%. The mean weight of hydropic foetuses was 1650 g and the gestation period at delivery 29 weeks. The rate of Caesarean sections in both groups was as high as 80%. Serious complications included thrombosis of the umbilicus, thromboembolism of the a. axillaris in a hydropic neonate and acute hypoxia of the foetus after transfusion. Four of six serious complications were recorded in hydropic foetuses.

[Successful completion of pregnancy in a dialyzed patient]. / Uspesne ukoncené tehotenství u dialyzované pacientky.

The authors describe the first case of a successful pregnancy in the Czech Republic in a patient treated by continuous ambulatory peritoneal dialysis. The 22-year-old patient became pregnant ten months after the onset of treatment by peritoneal dialysis. During pregnancy deterioration of arterial hypertension occurred, deterioration of anaemia and from the 29th week onwards cholestasis gravidarum developed. The development of the foetus was within normal limits. Because of suspected preeclampsia the pregnancy was terminated during the 35th week by Caesarean section. The patient was delivered of a healthy eutrophic boy without any congenital abnormalities.

[Is chorionic biopsy a dangerous and outdated method?]. / Je choriová biopsie nebezpecnou a prekonanou metodou?

The authors evaluated the risk of chorion biopsy used within the framework of prenatal diagnosis of the foetus. The incidence of abortions following transabdominal CVS (0.6%) did not differ from abortions after amniocentesis in the second trimester (0.5%). In a group of 1002 diagnostic biopsies of the chorion the authors did not record in the born infants reduction deformities of the extremities. Chorion biopsies were performed in 98% between the 10th and 12th week of gestation. The incidence of mosaicism--1.59%--was not significantly higher than reported in the literature. The authors recommend: to increase the size of the sample of chorion tissue by the use of a manual aspirator, the use of a 30 ml plastic syringe or the double needle method. They consider CVS a suitable method which serves the diagnosis of IUGR and the prenatal diagnosis of the foetus before the 20th week of gestation when cordocentesis involves risk.

The direct early diagnosis of cystic fibrosis by the detection of the delta F508 CFTR gene mutation in a prematurely delivered boy.

The suspicion of prenatal meconium ileus syndrome was raised in a pregnancy in a family with no history of cystic fibrosis because of significantly higher maternal serum alpha-fetoprotein in the 16th and 19th week of gestation, dispersed areas with increased echogenity in the fetal abdomen, slight fetal ascites in the 24th-25th weeks of gestation, decreased amniotic fluid gamma-glutamyltranspeptidase (GGT) activity and alpha-fetoprotein level in the 25th-26th weeks, and normal 46,XY karotype of the fetus. The detection of a homozygous deltaF508 cystic fibrosis transmembrane regulator (CFTR) gene mutation, by means of PCR from a small amount of white blood cells and urine sediment cells, substantiated the diagnosis of cystic fibrosis in a prematurely delivered boy in the 28th week of gestation. The repeated sweat test was unsuccessful. The autopsy examination confirmed the diagnosis of cystic fibrosis. Fetal meconium ileus syndrome was complicated by peritonitis and by formation of a meconium pseudocyst. Direct PCR typing improves postnatal diagnostic possibilities in the early neonatal period in prematurely delivered babies when the sweat test is difficult to perform.

[Biochemical monitoring of the fetoplacental unit in pregnant women with a hypotrophic fetus]. / Biochemické monitorování fetoplacentární jednotky tehotné zeny s hypotrofickým plodem.

The authors investigate the importance of examination of total urinary oestrogens (ET), oestriol (E3) trophoblast-specific beta-1-globulin (SP1) and alpha-1-antrypsin (A1AT) in the blood in a group of 56 pregnant women with suspection of a hypotrophic foetus. They made 192 analyses of ET, 144 E3; 153 SP1; 124 A1AT. With regard to the state of the neonate after delivery they divided the group into sub-groups with a eutrophic neonate and the group with a hypotrophic neonate, and the group with a hypotrophic neonate. They proved by statistical methods a different incidence of pathological values of ET, E3 and SP1 in the two sub-groups. A1AT rises in both sub-groups with advancing pregnancy. The incidence of pathological values was not significant. Assessment of ET is indispensible in any obstetric department; repeated examination is necessary, as one result does not provide adequate information on the function of the placenta and state of the foetus. To confirm the diagnosis, several biochemical parameters should be assessed.

The experience with the foetal diagnosis of the cystic fibrosis in the second and first trimester.

The amniotic fluid activity of gamma glutamyl transpeptidase (GGT), leucine aminopeptidase (LAP) and alcaline phosphatase (AP) and disacharidases was examined in 66 pregnancies with the risk of cystic fibrosis (CF) in the 17th-21st weeks of gestation. So far 28 pregnancies continue. The prenatal diagnosis was confirmed in all so far delivered children or aborted foetuses if the GGT activity was higher than 400 U/1 (10th percentile) or lower than 190 U/1 (3rd percentile) in the 17th-18th weeks. The results of other microvillar and ultrasound examinations were consistent with it. From 3 pregnancies with GGT activity in the range of 3-5 percentiles and abnormal activities of other microvillar enzymes, the CF was confirmed only in one aborted foetus with meconium ileus and with abnormal ultrasound examination. In other 2 pregnancies with normal ultrasound, healthy children were delivered. In 3 pregnancies with the GGT in the range of 5-10 percentiles and abnormal other microvillar enzymes, one false negative GGT and ultrasound examination was disclosed. The other 2 aborted foetuses did not exhibit the signs of CF in necropsy examinations. The meconium ileus was found in 2/4 of aborted foetuses with GGT lower than 3 percentiles, abnormal activities of other microvillar enzymes and abnormal ultrasound examination. The ultrasound examination was correct in 2/10 of pregnancies with GGT lower than 3 percentiles or abnormal activities of other microvillar enzymes. The GGT examination in 19th-21st weeks provided similarly reliable diagnostic results. The importance of fetal karyotyping and ultrasound elimination of other severe congenital anomalies is pointed out for critical interpretation of microvillar enzyme activities testing.(ABSTRACT TRUNCATED AT 250 WORDS)