OBJECTIVES:Implementation and introducing of first trimester screening using nuchal translucency (NT), nasal bone (NB), maternal serum pregnancy associated plasma protein-A and free beta-human chorionic gonadotropin (PAPP-A and fbeta-hCG) as a new method of screening for Down's Syndrome in a One-Stop-Clinic for Assessment of Risk for chromosomal abnormalities in the Czech Republic. To prove its effectiveness and make it part of our daily practice. Modify and/or withdraw the existing current practice.METHODS:Prospective study of 5010 singleton pregnancies who attended our clinic for prenatal screening for chromosomal and congenital abnormalities between 11-13+6 weeks gestation from January 2005 to December 2007. Pregnancies were followed up to ascertain live birth of Down syndrome babies.CONCLUSION:The uptake was 100% for screening and around 95% for invasive testing in the screen positive group. All expected trisomies 21 were detected along with many other aneuploidies for false positive rate of 4.4%. The study demonstrated the feasibility and efficacy of introducing OSCAR for chromosomal abnormalities in a University Hospital setting. The introduction of an examination of nasal bone reduced our false positive results and increased our sensitivity. Women who participated in our screening strategy were no longer advised to undergo the triple test.

OBJECTIVEMolecular techniques focused on detection of common aneuploidies--FISH and QFPCR--provide a quick result in prenatal diagnosis. There is a trend to apply these rapid tests as 'stand-alone' tests which would lead to substantial economical savings. The purpose of the retrospective study is to determine the frequency of chromosomal aberrations (CA) which would be missed by this tool in particular indication groups--residual cytogenetic risk.DESIGNRetrospective study.SETTINGDepartment of Medical Genetics and Foetal Medicine University Hospital Olomouc.METHOD5305 prenatal samples were examined and the frequency of QFPCR undetected CA (structural and rare aneuploidies) was observed.RESULTSThe residual risk in patients referred for abnormal results of current prenatal screening programs for Down syndrome or for maternal age, without any ultrasound (US) pathological findings, was 0.9%. It was 6.9% in the group where US pathological findings or family history of CA were present.CONCLUSIONThe rapid test can replace karyotyping if there is a risk for CA based exclusively on abnormal results of current screening programes for Down syndrome or age related risk, providing that US is normal.

AIM:The aim of this study was to evaluate TSPY (testis specific protein on the Y chromosome) gene and 5'UTR (UnTranslated Region) polymorphisms in men with impaired fertility compared to fertile controls.METHODS:We analyzed 72 infertile men and 31 fertile controls usingconventional sequencing analysis to find crucial SNPs (single nucleotide polymorphism) and other changes.RESULTS:The most remarkable changes were found in the 1(st) exon only. In one half of the both infertile men and fertile controls, the most frequent finding was 26 SNPs with a similar pattern. In the other half we found highly relevant changes, generating a stop codon in the first third of exon 1. Early termination cut down the protein by 78.5%. This kind of change was not found in the fertile controls. No correlation was found between the spermiogram and the changes leading to the stop codon. The distribution of men with deletions, insertion and higher gene copy number was not statistically different.CONCLUSION:The changes found in exon 1 in infertile men could fundamentally affect the process of spermatogenesis. These findings could significantly enhance our understanding of the molecular-genetic causes of male infertility.

Shifting our strategies of screening from second to first trimester in an OSCAR mannar and make it one of our daily practices. We present our experience in the University Hospital, Olomouc and the Regional Hospital Zlin. Our decision was based on a one year pilot study at the department and the published data in the field. Pregnant women were booked for in first trimester instead of the second and biochemical screening in the first trimester to substitute the triple test. Anomaly scan was organized for 20–22 weeks of pregnancy including screening of the heart. Minimum investment was made to purchase the software (Astraia) and a blood analyzer (Kryptor). If a women had a screen positive test we immediately progress to CVS and use PCR for the diagnosis, results are available in less than 36 hours. Confirmed pathological cases are terminated according to the wish of the women. We started the clinic in January 2004 in Olomouc and October 2007 in Zlin, we had about 18000 screening, we detected all 51 cases of trisomy 21 (except one) and 67 anther cases of other chromosomal abnormalities for 3.5% false positive rate. Shifting the strategy of prenatal screening required only a minimum investment in terms of money in a large district hospital, with a good use of existing resources including manpower. The new screening method proved a big success with high detection rates, low false positive rates and is very well accepted by women.

BACKGROUND Hemolytic disease in the newborn with its typical signs and poor prognosis has been known for centuries. Historically it can be divided into three pathological states which are fetal hydrops (hydrops fetus universalis), neonatal jaundice (icterus neonati gravis familiaris) and fetal anemia (anemia neonati). Almost 70 reports with quite accurate descriptions were found up to the end of 19th century. The patho physiological basis of the condition began to be studied at the beginning of the last century and the development of our knowledge is an example of the cooperation between pathologists, pediatricians, hematologists and later, obstetricians, immunologists and geneticists. Despite all the advances in this field it remains a serious disease up to this time. It is not managed successfully in all cases and despite successful immunological prophylaxis there are cases when we need to administer intrauterine transfusion based on the information received by dopplerometric measurement of arteria cerebri perfusion and fetal blood sampling. METHODS Review of lover cited literature. CONCLUSION The history of the hemolytic disease in the newborn, its condition and approaches to it has not been recently compiled in the Czech Republic.

AIM OF STUDY:To assess cell free fetal DNA (cffDNA) fragmentation rate in pregnant women during the course of gravidity.STUDY DESIGN:QF PCR efficiency in cffDNA and quantitative analyses in particular cffDNA molecular size fractions.SETTING:The study was performed at Department of Medical Genetics and Fetal Medicine, University Hospital Olomouc.METHOD:1. 363 plasma DNA samples from women in different week of pregnancy (from 4th w.g. to 37th w.g.) were tested for QF PCR efficiency in particular STRs and AMELX/Y. 2. Size fractionated cff DNA (150-300 bp, 300-500 bp, 500-760 bp) was quantified by QF PCR in 91 pregnant women (from 9th w.g. to 40th w.g.). 3. Size fractionated cff DNA from male fetuses was quantified by real time PCR (SRY/internal control) in 22 pregnant women (from 9th w.g. to 36th w.g.).RESULTS:1. QF PCR efficiency decreased from longer to shorter molecules. 2. The only 500 -760 bp fraction showed cffDNA increase in relation to week of gravidity. 3. Indirect relation between amount of cffDNA and week of gravidity was found in 150-300 bp fraction by Real-time PCR.CONCLUSION:Assembling of all 3 approaches indicates increase of longer cffDNA molecules during the gravidity while level of the short cffDNA molecule fragments probably remains from the approximately 9th w.g. the same.

BACKGROUND:About 20 percent of the population in developing countries is composed of women of reproductive age. These women face one of the catastrophic risks of pregnancy "uterine rupture". Studies conducted in the developing world give strong evidence that uterine rupture is a major health problem in these countries with the rate being high in rural areas.AIM:The purpose of the study was to estimate the incidence and determine the risk factors and outcome of uterine rupture among women using the referral hospital Al-thawra in Sana'a City, Yemen republic and to extrapolate the data to the whole of Yemen.METHODS:The data was collected retrospectively; by interviewing, examining and following up all the cases of uterine rupture coming to the hospital during a period of 9 months between September 1996 and May 1997. A descriptive analysis and distribution frequency of the commonest causes of uterine rupture in 37 cases are presented taking into account medical, reproductive, health services provided and sociodemographic factors.RESULTS:Incidence of uterine rupture in Yemen was found to be (0.63), obstructed labor 83 %, contracted pelvis 19 %, previous surgery in 48 %, Oxytocine infusion in 42 %. Grand-multiparty was in 65 % and maternal age over 35 years in 50 %. Antenatal care was only in 34 %.CONCLUSION:The high percentage of malpresentation, cephalopelvic disproportion, previous uterine surgery accompanied by the high percentage of use of Oxytocin in this study highlights very clearly the role of this medication in increasing the risk of uterine rupture in Yemen.

The goal of the study is to inform about clinical and genetic aspects of tuberuos sclerosis, to point out the benefit of genetic testing as well as its limits, and to inform about the optimised genetic testing procedure. Tuberous sclerosis is an autosomally dominant hereditary neurocutaneous disease with extremely variable clinical manifestations. The disease results from mutation in one of the two tumour-suppressor genes, i.e. TSC1 or TSC2. A wide range of causal mutations with random distribution in the responsible genes has been described. The proposed DNA examination procedure was designed on the basis of knowledge obtained from the analysis of 65 samples from Czech patients and by tests performed on 53 positive control samples from partner centres abroad. The basic criteria for the selection of the best procedure for DNA diagnosis in TSC patients was economy, efficiency and laboratory capacity.

Cell‐free fetal (cff) DNA analysis by short tandem repeats (STR) has the advantage of better recognizing the different genotypes. However, quantitative examination by quantitative fluorescent (QF) polymerase chain reaction (PCR) by STRs is limited to only a rough approximation. This project focuses on a more precise calculation of the relative cff DNA amount tested in the STRs' loci.

OBJECTIVE:Objective of our study was optimization of noninvasive fetal sex detection from maternal plasma in pregnant women.STUDY DESIGN:Molecular DNA quantitative analyses in gonosomal loci.SETTING:The study was performed at Department of Medical Genetics and Fetal Medicine, University Hospital Olomouc.METHODS:Together 475 DNA samples isolated from maternal plasma in different weeks of pregnancy ranging from 4th w.g. to 37th w.g. Y chromosomal sequences in AMELY and TSPY were tested by refined quantitative fluorescent PCR using capillary electrophoresis.RESULTS:The method is able to distinguish 1% of Y chromosomal sequences of artificial mixtures. Investigation and assessment in cell free fetal DNA samples achieved 4.05% of false positivity and 7.15% of false negativity in Y sequence detection.CONCLUSION:Established method allows detecting fetal sex with high sensitivity and specificity. The method is possible to use also for quantitative purposes.

Paternity testing is nowadays mostly based on the analysis of DNA short tandem repeats (STR). Number and selection of STR loci differ according to identification kit producers and also particular laboratories. This article refers a revision of formerly excluded paternity. The cause of the mismatch was revealed by the enlargement of STR loci panel and reciprocal evaluation of samples and paternity was on the contrary confirmed. Different possibilities of failures, their consequences and preventions are also discussed.

To determine the possible association between single umbilical artery (SUA) in the second trimester of pregnancy and the incidence of chromosomal abnormalities. To determine whether the presence of chromosomal defects in fetuses with SUA is related to the side of the missing artery.

OBJECTIVES:The aim of this study is to evaluate the significance of nasal bone as a marker for trisomy 21 in the group of women that underwent invasive procedures in our center at 11 to 14 weeks' gestation.METHODS:The data of 181 women who had undergone the invasive procedures were evaluated for the presence or absence of nasal bone retrospectively and were correlated with fetal karyotype.RESULTS:A successful view of the fetal profile was obtained in 135 fetuses. The nasal bone was absent in 5 of 8 fetuses with trisomy 21 and in 3 of 4 fetuses with trisomy 18. In the group of chromosomally normal fetuses the nasal bone was absent in 4 of the 123 cases. The false positive rate of our screening program dropped from 4.5% to 2.5% after the introduction of the nasal bone evaluation into our risk calculation model for trisomy 21.CONCLUSION:Nasal bone evaluation improved the detection of trisomy 21 in the first trimester in our screening program and reduced the need for invasive procedures in our department. Absence of the nasal bone showed a sensitivity of 63% for a 5% false positive rate for trisomy 21 in our study. It is rarely observed in chromosomally normal fetuses (2.5%). An appropriate training is mandatory in order to achieve acceptable results.

The human TSPY (testis-specific protein, Y-linked) gene family (30-60 copies) is situated in the MSY (male-specific) region of the Y chromosome. Testis-specific expression indicates that the gene plays a role in spermatogenesis. Refined quantitative fluorescence PCR (polymerase chain reaction) was applied to evaluate the relative number of TSPY copies compared with AMELY/X (amelogenin gene, Y-linked) genes in 84 stratified infertile men and in 40 controls. A significantly higher number of TSPY copies was found in infertile men compared with the controls (P = 0.002). The diagnostic discrimination potential of the relative number of TSPY copies was evaluated by receiver operating characteristic curve analysis. TSPY/AMELY was unambiguously found to be powerful in the diagnostic separation of both the control samples and the infertile men, reaching a good level of specificity (0.642) and sensitivity (0.732) at a cut-off point of 0.46. The findings were supported by independently repeated studies of randomly selected positive samples and controls. Evaluation of the TSPY copy number offers a completely new diagnostic approach in relation to the genetic cause of male infertility. The possible effect of the copy number of TSPY genes on spermatogenesis may explain indiscrete pathological alterations of spermatid quality and quantity.