The aim of the study was to evaluate the effectiveness of placental-specific markers, extracellular fetal DNA (sex-determining region Y and hypermethylated RASSF1A sequences) and circulating C19MC microRNAs (miR-516-5p, miR-517-5p, miR-518b, miR-520a-5p, miR-520h, miR-525, and miR-526a) for the diagnosis and consecutive follow-up of gestational trophoblastic disease/neoplasia. Increased levels of extracellular fetal DNA and C19MC microRNAs were detected in patients with active disease when compared with the period when the patients reached remission of the disease. The positive correlation between plasma levels of hypermethylated RASSF1A sequence, C19MC microRNAs, and human chorionic gonadotropin serum levels was found. MiR-520a-5p had the best performance to detect patients with active disease (a positive predictive value of 100% at a null false positive ratio (FPR)). MiR-516-5p and miR-525 were able to diagnose 100% of women with active disease at the FPR 3.9%/7.7%. The overall predictive capacity of single miR-526a (81.8% at null FPR), miR-517-5p (90.9% at 15.4% FPR), miR-518b (100% at 38.5% FPR), and miR-520h (90.9% at 26.9% FPR) biomarkers to detect active disease cases was slightly lower. Transient increase in C19MC microRNA plasma levels after the first cycle of chemotherapy indicated the decay of placental trophoblast residual tissue. The increased levels of extracellular fetal DNA and placental-specific C19MC microRNAs are associated with gestational trophoblastic disease/neoplasia. Screening of extracellular placental-specific biomarkers may represent an additional option to identify a significant proportion of women with active disease and to monitor the therapy response. Non-invasive follow-up of the decomposing residual tissue in the form of extracellular nucleic acids of placental origin packed into apoptotic bodies derived from placental trophoblasts is available.

• MeSH
• choriogonadotropin krev   MeSH
• DNA krev   MeSH
• dospělí MeSH
• gestační trofoblastická nemoc krev   diagnóza   genetika   MeSH
• lidé středního věku MeSH
• lidé MeSH
• metylace DNA * MeSH
• mikro RNA krev   MeSH
• mladiství MeSH
• nádorové supresorové proteiny genetika   MeSH
• následné studie MeSH
• protein oblasti určující pohlaví na chromozomu Y genetika   MeSH
• těhotenství MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

We determined the feasibility of universal fetal marker detection in maternal circulation. Using real-time PCR, we compared the levels of fetal (SRY and hypermethylated RASSF1A) and total (GLO gene and total RASSF1A) extracellular DNA and fractions of extracellular fetal DNA (SRY/GLO vs. hypermethylated RASSF1A/total RASSF1A) in maternal circulation. Sensitivity and specificity reached 100% as the fetal-specific hypermethylated RASSF1A sequence was detected in all 151 examined plasma samples derived from 70 normal pregnancies with a singleton male (n=51) or female (n=19) fetus sampled throughout gestation and absent in non-pregnant individuals (n=29). A strong positive correlation was observed between fetal-derived hypermethylated RASSF1A and SRY (ρ=0.66, P<0.001), total RASSF1A and GLO (ρ=0.65,P<0.001), SRY/GLO vs. hypermethylated RASSF1A/total RASSF1A ratio (ρ=0.62, P<0.001) in maternal plasma. The results indicate that a universal fetal marker could be useful not only for the confirmation of the presence of fetal cell-free DNA in maternal plasma but could enable quantification of cell-free fetal DNA in pregnancy associated disorders, independently of the sex of the fetus.

• MeSH
• alkoholoxidoreduktasy krev   genetika   MeSH
• DNA krev   MeSH
• lidé MeSH
• metylace DNA genetika   MeSH
• nádorové supresorové proteiny krev   genetika   MeSH
• plod MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• prenatální diagnóza metody   MeSH
• protein oblasti určující pohlaví na chromozomu Y krev   genetika   MeSH
• retrospektivní studie MeSH
• senzitivita a specificita MeSH
• těhotenství krev   genetika   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• těhotenství krev   genetika   MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

This study evaluated quantification of fetal extracellular DNA in maternal plasma for differentiation between cases at risk of onset of placental-insufficiency-related complications and normal pregnancies. Using real-time polymerase chain reaction, fetal (sex-determining region Y [SRY] and hypermethylated RASSF1A sequence) and total (beta-globin [GLO] gene) extracellular DNA was examined in 70 normal pregnancies, 18 at risk of placental-insufficiency-related pregnancy complications, 24 preeclampsia with or without (w or w/o) intrauterine growth retardation (IUGR) (median 34.0 week), and 11 IUGR (median 28.5 week). IUGR was diagnosed when estimated fetal weight was below the 10th percentile for evaluated gestational age. Although increased levels of extracellular DNA were detected in pregnancies with preeclampsia w or w/o IUGR relative to controls (RASSF1A, p < 0.001; SRY, p = 0.009; GLO, p < 0.001), quantities of fetal extracellular DNA in IUGR were not statistically significant (RASSF1A, p = 0.21; SRY, p = 0.2). RASSF1A, SRY, and GLO achieved 93.1%, 93.6%, and 92.1% accuracy for differentiation between normal pregnancy and preeclampsia w or w/o IUGR. Lower sensitivity was observed for pregnancies with onset of IUGR (RASSF1A, 60.0%; SRY, 80.0%; GLO, 72.7%), but did not influence final accuracy (RASSF1A, 91.6%; SRY, 92.5%; GLO, 89.5%). Among 18 patients at risk, 8 pregnancies involving 3 female and 5 male fetuses developed preeclampsia (n = 4), IUGR (n = 3), and chronic placentopathy causing hypoxia (n = 1). Elevation of extracellular DNA was demonstrated in 3/5 (SRY), 1/8 (hypermethylated RASSF1A), and 4/8 (GLO) patients at the earliest 26 weeks and at the latest 2 weeks before the onset of symptoms. These data indicate that fetal and total extracellular DNA concentrations can be significantly elevated in plasma of patients who later developed placental-insufficiency-related pregnancy complications. However, this is strongly individualized, and not a rule for all cases, and probably depends on the actual occurrence of excessive placental trophoblast apoptosis.

• MeSH
• beta-globiny genetika   metabolismus   MeSH
• časové faktory MeSH
• DNA genetika   MeSH
• extracelulární prostor genetika   MeSH
• genetické markery genetika   MeSH
• lidé MeSH
• matky MeSH
• metylace DNA MeSH
• nádorové supresorové proteiny krev   genetika   MeSH
• placentární insuficience krev   diagnóza   genetika   MeSH
• plod cytologie   MeSH
• protein oblasti určující pohlaví na chromozomu Y krev   genetika   MeSH
• riziko MeSH
• studie případů a kontrol MeSH
• těhotenství MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Poruchy pohlavního vývoje se vyskytují přibližně u 1:4500 živě narozených dětí. Jejich etiologie je velmi pestrá a dosud ne zcela objasněná. S přibývajícími poznatky vyvstala potřeba nové klasifikace a nomenklatury, které byly navrženy jako takzvaný Chicagský konsensus v roce 2005. Velkou pozornost v současné době vzbuzují poruchy pohlavního vývoje, jejichž podstatou jsou mutace genů řídicích vývoj gonád. Ve vyvíjejících se pohlavních žlázách se exprimuje přes 200 genů; u deseti z nich byl již popsán vliv patogenníchmutací nebo neadekvátního počtu genových kopií na poruchy pohlavního vývoje u člověka. Přesné určenímolekulárně- genetické podstaty těchto poruch umožní zpřesnit diagnózu, dopad na následný pohlavní vývoj a genetická rizika pro další členy rodiny.

The disorders of sex development occur in about 1:4,500 live born children. Their etiology is varied and not yet clearly understood. As the knowledge increased the need of a new classification has become apparent and the nomenclature was recommended as a so called Chicago Consensus in 2005. Considerable attention is presently devoted to disorders in sex development, which are based on mutations of genes controlling gonadal development. In the developing gonads about 200 genes are expressed; in ten of them the effects of pathogenic mutations or inadequate gene number copies on the human sexual development have already been described. A precise determination of molecular-genetic basis will make a more precise diagnosis possible as well as the elucidation of possible impact on subsequent sexual development and genetic risk for other members of the family.

• MeSH
• alfa-talasemie genetika   MeSH
• dítě MeSH
• geny sry fyziologie   genetika   MeSH
• klasifikace metody   MeSH
• lidé MeSH
• mutace genetika   MeSH
• pohlavní orgány abnormality   patofyziologie   patologie   MeSH
• poruchy gonád diagnóza   genetika   patologie   MeSH
• protein oblasti určující pohlaví na chromozomu Y genetika   MeSH
• proteiny hedgehog genetika   MeSH
• proteiny Wnt genetika   MeSH
• proteiny WT1 genetika   MeSH
• steroidogenní faktor 1 genetika   MeSH
• vrozené, dědičné a novorozenecké nemoci a abnormality genetika   MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• Publikační typ
• přehledy MeSH

The aims of our research involved to investigate DYS-14 copy number variations in healthy males, to quantify extracellular DNA in maternal circulation in normal versus complicated pregnancies, and to study variations in the DYS-14 copy number in extracellular male fetal DNA. Fifty-five healthy males, 43 uncomplicated male singleton pregnancies (23 sampled at the 16th week and 20 sampled at the 36th week), and 15 pregnancies with placental insufficiency (PI)-related complications (mean 34.1 weeks) were analyzed using real-time PCR with DYS-14 sequence, sex determining region Y (SRY), and beta-globin (GLO) genes used as markers. Increased levels of extracellular DNA were detected in PI-related complications relative to gestational age-matched controls (SRY, p < 0.001; DYS-14, p = 0.007; GLO, p < 0.001). When the mean + 2SD (standard deviation) of controls was used as a cutoff, SRY, DYS-14, and GLO achieved 91.7%, 68.8%, and 94.4% accuracy, respectively, for differentiation between normal and complicated pregnancies. Considerable variations in the DYS-14 copy number in healthy males (mean 52.6) and extracellular DNA were found. A lower DYS-14 copy number was observed in PI-related complications (mean 83.5) compared to uncomplicated pregnancies (16th week: mean 114.2, p = 0.02; 36th week: mean 142.8, p = 0.04). The DYS-14 copy number was higher in extracellular DNA throughout gestation relative to healthy males. We concluded that, regarding interindividual copy number variations, the DYS-14 sequence is not an optimal marker for extracellular fetal DNA quantification for differentiation between normal and complicated pregnancies.

• MeSH
• dítě MeSH
• DNA krev   MeSH
• dospělí MeSH
• genová dávka MeSH
• gestační stáří MeSH
• kojenec MeSH
• lidé středního věku MeSH
• lidé MeSH
• lidský chromozom Y genetika   MeSH
• matky MeSH
• mladiství MeSH
• mladý dospělý MeSH
• placentární insuficience diagnóza   genetika   MeSH
• plod MeSH
• předškolní dítě MeSH
• prenatální diagnóza MeSH
• protein oblasti určující pohlaví na chromozomu Y genetika   MeSH
• proteiny buněčného cyklu MeSH
• studie případů a kontrol MeSH
• těhotenství MeSH
• výsledek těhotenství MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• kojenec MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• práce podpořená grantem MeSH
• srovnávací studie MeSH

The transplacental cell transfer naturally takes place during pregnancy and occurs bi-directionally between the mother and fetus. Using real-time polymerase chain reaction (PCR) assay and sex determining region Y (SRY) gene as a marker, we examined the presence of male fetal cells in cell cultures derived from synovial tissues and skin dermis in women with prior pregnancy history suffering from rheumatoid arthritis (RA) who underwent synovectomy. Male DNA was detected in synovial cell samples derived from carpal, hip, metacarpophalangeal and metatarsophalangeal joints in five out of 13 (38.5%) patients with RA in a frequency range of 0.02-62.55 (mean 12.17) male cells per 10,000,000 total cells. SRY gene positivity was found as well in skin fibroblast cultures in four out of 10 (40.0%) RA patients in a frequency range of 3.26-43.47 (mean 15.42) male cells per 10,000,000 total cells, respectively. The difference in a frequency of fetal-derived male cells between both the cohorts did not achieve the statistical difference (p=0.77). We conclude that persisting male fetal cells are able to grow from non-inflamed tissues as well as from those which have many features characteristic of a stressed tissue. We conclude that persisting male fetal cells are also able to proliferate in cell culture since their presence was detected even in consecutive passages.

• MeSH
• chimérismus MeSH
• DNA analýza   MeSH
• dospělí MeSH
• fibroblasty chemie   MeSH
• genetické markery genetika   MeSH
• kultivované buňky MeSH
• kůže cytologie   MeSH
• lidé MeSH
• mezoderm cytologie   růst a vývoj   MeSH
• mladý dospělý MeSH
• plod cytologie   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• protein oblasti určující pohlaví na chromozomu Y genetika   MeSH
• revmatoidní artritida komplikace   MeSH
• synovektomie MeSH
• synoviální membrána cytologie   chemie   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• práce podpořená grantem MeSH