Metoda fluorescenční in situ hybridizace (FISH) umožňuje stanovit frekvenci chromozomálně abnormálních spermií v ejakulátu. V minulosti byly publikovány základní hladiny disomií pro jednotlivé chromozomy (0,03–0,47 %) zjištěné metodou FISH ve spermiích normálních zdravých dárců. Celkově je u zdravých mužů asi 7 % spermií chromozomálně abnormálních, nejčastější jsou aneuploidie chromozomů 21, X a Y. Byla však zaznamenána významná individuální variabilita. Vyšší frekvence numerických aberací se často nacházejí v ejakulátu mužů s abnormálním spermiogramem, u nositelů vrozených chromozomálních abnormalit (numerické aberace pohlavních chromozomů, marker chromozomy, balancované translokace a inverze) a ve spermiích získaných TESE. Největší přínos má vyšetření spermií metodou FISH pro nositele vrozených reciprokých a Robertsonových translokací, příp. pericentrických inverzí. Publikované frekvence chromozomálně nebalancovaných spermií se pohybují v rozmezí 0–37 % u nositelů pericentrických inverzí, 3–40 % u nositelů Robertsonových translokací a až 23–81 % u nositelů reciprokých translokací. Stanovení frekvence chromozomálně abnormálních spermií umožňuje individualizaci rizika u jednotlivých pacientů, informované rozhodování páru při plánování rodičovství a predikci úspěšnosti IVF cyklu.

The fluorescence in situ hybridization (FISH) method allows detection of frequencies of chromosomally abnormal spermatozoa in semen. Baseline levels of disomies for different chromosomes (0.03–0.47%) detected by FISH method in sperm from normal healthy donors have previously been published. Semen of healthy men usually contains approximately 7% of chromosomally abnormal spermatozoa, with aneuploidies of chromosomes 21, X and Y being the most frequent. However, a significant inter-individual variability was observed. Higher frequencies of numerical aberrations are often found in men with abnormal semen characteristics, in carriers of congenital chromosomal abnormalities (numerical sex chromosome aberrations, marker chromosomes, balanced translocations and inversions) and in sperm obtained by TESE. Sperm evaluation by the FISH method is of benefit especially for carriers of congenital reciprocal and Robertsonian translocations, or of some pericentric inversions. Published frequencies of chromosomally unbalanced sperm range from 0–37% in carriers of pericentric inversions, 3–40% in carriers of Robertsonian translocations and even 23–81% in carriers of reciprocal translocations. Assessment of frequencies of chromosomally abnormal spermatozoa allows personalized risk estimation in individual patients, making informed decisions concerning family planning and the prediction of IVF cycle outcome.

• Klíčová slova
• FISH, chromozomové aberace, balancované translokace, inverze,
• MeSH
• chromozomální aberace klasifikace   MeSH
• financování organizované MeSH
• hybridizace in situ fluorescenční metody   využití   MeSH
• lidé MeSH
• spermie cytologie   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH

A family pilot study was conducted in the Czech Republic to test the hypothesis that exposure to air pollution with particulate matter (PM) in children results in detectable effects indicated by a number of biomarkers of exposure and early effects. The frequency of micronuclei (MN) in peripheral blood lymphocytes (PBLs) was analysed to assess the cytogenetic effects in children and mothers living in two different areas. From each area two groups of children from a total of 24 families (mean age: 6.0+/-0.6 and 9.0+/-1.2 years) in a total of 47 children and 19 mothers (mean age: 33.6+/-3.9 years) participated. Chromosome aberrations determined with fluorescence in situ hybridization (FISH) painting for chromosomes #1 and #4 were analysed in 39 children and 20 parents. Teplice, a mining district, in Northern Bohemia was selected for the analyses of the effects in a population exposed to high levels of air pollution, especially during winter, and compared with a population from the rural area of Prachatice in Southern Bohemia. Significant higher frequencies of MN were found in the younger children living in the Teplice area as compared with those living in the Prachatice area (7.0+/-2.3 per thousand versus 4.9+/-2.0 per thousand, p=0.04). Higher levels of MN were also measured in the older children and the mothers from the Teplice area (9.2+/-3.7 per thousand versus 6.6+/-4.4 per thousand) and (12.6+/-3.4 per thousand versus 10.1+/-4.0 per thousand). The increased MN frequency may be associated with elevated carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) concentration of the PM(2.5) measured in the ambient Teplice air, but other factors like genotoxic compounds from the diet or protective effect of micronutrients, which was not addressed in this pilot study, may also differ between the two areas. MN frequencies were found to increase with age in children. Lower MN frequency was found in boys as compared to girls. The result of the FISH analyses showed a low number of individuals with detectable levels of aberrations and no significant increases in genomic frequency of stable chromosome exchanges (F(G)/100) were found in children or parents from the Teplice area in comparison with those from the Prachatice area. The family pilot study indicates that MN is a valuable and sensitive biomarker for early biological effect in children and adults living in two different areas characterised with significant exposure differences in c-PAHs concentrations during winter.

• MeSH
• cytogenetika MeSH
• dítě MeSH
• dospělí MeSH
• hybridizace in situ fluorescenční MeSH
• látky znečišťující vzduch toxicita   MeSH
• lidé MeSH
• mikrojaderné testy MeSH
• mikrojádra chromozomálně defektní * chemicky indukované   MeSH
• pilotní projekty MeSH
• předškolní dítě MeSH
• sourozenci MeSH
• vystavení vlivu životního prostředí MeSH
• znečištění ovzduší škodlivé účinky   MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

• MeSH
• aneuploidie MeSH
• hybridizace in situ fluorescenční využití   MeSH
• oocyty fyziologie   ultrastruktura   MeSH
• prasata MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH

To examine interindividual differences in sperm chromosome aneuploidy, repeated semen specimens were obtained from a group of ten healthy men, aged 20-21 at the start of the study, and analyzed by multi-color fluorescence in situ hybridization (FISH) analysis to determine the frequencies of sperm aneuploidy for chromosomes X, Y, 8, 18 and 21 and of diploidy. Semen samples were obtained three times over a five-year period. Statistical analysis examining the stability of sperm aneuploidy over time by type and chromosome identified two men who consistently exhibited elevated frequencies of sperm aneuploidy (stable variants): one with elevated disomy 18 and one with elevated MII diploidy. Differences among frequencies of aneuploidy by chromosome were also seen. Overall, disomy frequencies were lower for chromosome X, 8 and 18 than for chromosomes 21 or Y and for XY aneuploidy. The frequency of chromosome Y disomy did not differ from XY sperm frequency. Also, the frequency of meiosis I (XY) and II (YY + XX) sex chromosome errors did not differ in haploid sperm, but the frequency of MII errors was lower than MI errors in diploid sperm. Frequencies of sperm aneuploidy were similar between the first sampling period and the second, two years later. However, the frequency of some types of aneuploidy (XY, disomy Y, disomy 8, total autosomal disomies, total diploidy, and subcategories of diploidy) increased significantly between the first sampling period and the last, five years later, while others remained unchanged (disomy X, 21 and 18). These findings confirm inter-chromosome differences in the frequencies of disomy and suggest that some apparently healthy men exhibit consistently elevated frequencies of specific sperm aneuplodies. Furthermore, time/age-related changes in sperm aneuploidy may be detected over as short a period as five years in a repeated-measures study. Copyright 2005 S. Karger AG, Basel.

• MeSH
• aneuploidie MeSH
• dospělí MeSH
• financování organizované MeSH
• hybridizace in situ fluorescenční MeSH
• lidé MeSH
• motilita spermií MeSH
• počet spermií MeSH
• sperma cytologie   fyziologie   MeSH
• spermie cytologie   fyziologie   MeSH
• uniparentální disomie genetika   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH

The efficient detection of chromosomal aberrations in childhood acute leukaemias presents a significant component in the diagnostics of this frequent malignant disease. We used comparative genomic hybridization (CGH) and high-resolution comparative genomic hybridization (HR-CGH) to determine the frequency of chromosomal changes in 33 children with acute leukaemia (AL). The yields of chromosomal abnormalities were compared with the results obtained using conventional cytogenetics (G-banding) and fluorescence in situ hybridization (FISH). Conventional cytogenetics revealed chromosomal changes in 17 (52 %) of studied patients. The employment of FISH together with G-banding analysis identified chromosomal changes in 27 (82 %) of the AL patients investigated. CGH detected changes in DNA copy numbers in 24 (73 %) patients, 40 losses and 67 gains were found in total. HR-CGH disclosed 98 losses and 97 gains in 26 (79 %) patients. In comparison with CGH, HR-CGH analyses unveiled 88 new chromosomal aberrations: 58 losses and 30 gains. The most commonly gained chromosomes were 21 (22.5 %), X (15 %), 18 (12,5 %) and 17 (10 %). The most common losses involved sub-regions or arms of chromosomes 7 (15 %), 9 (12.5 %), 16, 19 and 1 (10 % each). Cytogenetic and molecular cytogenetic analyses of 33 childhood acute leukaemias revealed chromosomal changes in total 31 (94 %) patients. The evaluation of HR-CGH sensitivity proved that the minimal cell population of malignant cells in which a certain chromosomal change could be found was close to the 20 - 30 % level. Our results confirm the benefits of HR-CGH in detecting chromosomal changes in childhood AL. Supplementing G-banding and FISH with the HR-CGH diagnostic method increases the detection of unbalanced structural chromosomal rearrangements and can reveal small cell clones with gains and losses of whole chromosomes in hyperdiploid AL.

• MeSH
• akutní nemoc MeSH
• chromozomální aberace MeSH
• dítě MeSH
• financování organizované MeSH
• hybridizace in situ fluorescenční MeSH
• hybridizace nukleových kyselin metody   MeSH
• kojenec MeSH
• leukemie genetika   MeSH
• lidé MeSH
• mladiství MeSH
• předškolní dítě MeSH
• pruhování chromozomů MeSH
• Check Tag
• dítě MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• hodnotící studie MeSH
• srovnávací studie MeSH

Data on the frequency of aneuploidy in farm animals are lacking and there is the need for a reliable technique which is capable of detecting all chromosomes simultaneously in a single cell. With the employment of comparative genomic hybridization coupled with the whole genome amplification technique, this study brings new information regarding the aneuploidy of individual chromosomes in pigs. Focus is directed on in vivo porcine blastocysts and late morulas, 4.7% of which were found to carry chromosomal abnormality. Further, ploidy abnormalities were examined using FISH in a sample of porcine embryos. True polyploidy was relatively rare (1.6%), whilst mixoploidy was presented in 46.8% of embryos, however it was restricted to only a small number of cells per embryo. The combined data indicates that aneuploidy is not a prevalent cause of embryo mortality in pigs.

• MeSH
• aneuploidie MeSH
• blastocysta cytologie   metabolismus   fyziologie   MeSH
• embryo savčí MeSH
• gestační stáří MeSH
• nemoci prasat diagnóza   embryologie   genetika   MeSH
• oocyty cytologie   metabolismus   fyziologie   MeSH
• prasata genetika   MeSH
• srovnávací genomová hybridizace metody   veterinární   MeSH
• těhotenství MeSH
• ztráta embrya diagnóza   genetika   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH

BACKGROUND: Alanced translocations are associated with infertility, spontaneous abortions and birth defects. METHODS: We report the analysis, by multicolour fluorescence in situ hybridization (FISH), of meiotic segregation and aneuploidy of chromosomes X, Y, 7, 8 and 21 in sperm from three men who are carriers of two different translocations involving chromosomes 11 and 18. A control group comprised ten young, healthy normospermic men. RESULTS: The higher prevalence of alternate segregation followed by adjacent 1, adjacent 2 and 3:1, and other segregants was observed in all three patients. Two carriers of the same translocation differed only in the frequency of adjacent 2 segregation (P < 0.01). The carrier of the other translocation showed significantly higher frequency of alternate (P < 0.01) and less adjacent 1 and 3:1 segregation products (P < 0.01). An increased frequency of XY (P < 0.01), YY (P < 0.05) and diploid (P < 0.01) sperm was also detected in the group of translocation carriers compared with the control group. This difference was caused by elevated frequencies of disomy and diploidy in two of our carriers. CONCLUSIONS: The incidence of chromosomally unbalanced or aneuploid gametes varies in the individual translocation carriers even if the same chromosomes are included in the translocation. FISH analysis provides information useful for genetic counseling and assisted reproduction.

• MeSH
• aneuploidie MeSH
• dospělí MeSH
• financování organizované MeSH
• hybridizace in situ fluorescenční MeSH
• lidé MeSH
• lidské chromozomy X genetika   MeSH
• lidské chromozomy, pár 11 genetika   MeSH
• lidské chromozomy, pár 18 genetika   MeSH
• lidské chromozomy, pár 21 genetika   MeSH
• lidské chromozomy, pár 7 genetika   MeSH
• lidské chromozomy, pár 8 genetika   MeSH
• lidský chromozom Y genetika   MeSH
• meióza fyziologie   MeSH
• segregace chromozomů MeSH
• spermie cytologie   MeSH
• translokace genetická genetika   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH

PROBLEM: The aim of this study was to investigate frequencies of eight antiphospholipid antibodies (aPLs) in serum, four genetic thrombophilic factors and their mutual relation in 206 patients with repeated pregnancy loss (RPL). METHOD OF STUDY: Enzyme-linked immunosorbent assay was used for detection of aPLs against ph-serine, ph-ethanolamine, ph-inositol, DL-glycerol, phosphatidic acid, anti-annexin V, cardiolipin, and beta2-GPI. FV 1691G>A (Leiden mutation), FII 20210G>A mutation, MTHFR 677C>T and MTHFR 1298A>C variant genotypes were determined using a melting curve analysis of the PCR amplification product detected by the fluorescence resonance energy transfer. Genotypic distribution and allelic frequencies were calculated. Correlation between aPLs and thrombophilic factors was tested by chi-square and Fisher exact test. RESULTS: Our results show significantly increased prevalence of aPLs against ph-inositol (17-19.6% dependent on number of spontaneous miscarriages) and against ph-serine (18-25%). aPLs in IgG prevail. In 96% of the studied group, at least one risk factor was found (either aPLs positivity or thrombophilic factor). Both aPLs and thrombophilic factors were present in 43%. In the group of women with three or more RPLs, strong positive correlation of aPLs positivity and thrombophilic risk factors was observed. CONCLUSION: Antiphospholipide antibodies and genetic thrombophilic factors are important risk factors in the pathogenesis of RPL. Both autoantibodies against various kinds of phospholipides and genetic thrombophilic factors must be studied together in diagnosis of RPL for appropriate treatment

• MeSH
• antifosfolipidové protilátky biosyntéza   genetika   imunologie   krev   MeSH
• DNA sondy MeSH
• dospělí MeSH
• ELISA MeSH
• frekvence genu imunologie   MeSH
• genotyp MeSH
• habituální potrat epidemiologie   genetika   imunologie   MeSH
• kauzalita MeSH
• lidé středního věku MeSH
• lidé MeSH
• mutace MeSH
• průřezové studie MeSH
• rizikové faktory MeSH
• těhotenství MeSH
• trombofilie genetika   imunologie   MeSH
• výsledek těhotenství MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

Complex chromosomal rearrangements (CCR) represent rare structural chromosome abnormalities frequently associated with infertility. In this study, meiotic segregation in spermatozoa of an infertile normospermic carrier of a 4-breakpoint t(1;3;6) CCR was analysed. A newly developed array comparative genomic hybridization protocol was used, and all chromosomes in 50 single sperm cells were simultaneously examined. Three-colour FISH was used to analyse chromosome segregation in 1557 other single sperm cells. It was also used to measure an interchromosomal effect; sperm chromatin structure assay was used to measure chromatin integrity. A high-frequency of unbalanced spermatozoa (84%) was observed, mostly arising from the 3:3 symmetrical segregation mode. Array comparative genomic hybridization was used to detect additional aneuploidies in two out of 50 spermatozoa (4%) in chromosomes not involved in the complex chromosome rearrangement. Significantly increased rates of diploidy and XY disomy were found in the CCR carrier compared with the control group (P < 0.001). Defective condensation of sperm chromatin was also found in 22.7% of spermatozoa by sperm chromatin structure assay. The results indicate that the infertility in the man with CCR and normal spermatozoa was caused by a production of chromosomally unbalanced, XY disomic and diploid spermatozoa and spermatozoa with defective chromatin condensation.

• MeSH
• analýza jednotlivých buněk MeSH
• body zlomu chromozomu * MeSH
• dospělí MeSH
• genová přestavba * MeSH
• heterozygot MeSH
• hybridizace in situ fluorescenční MeSH
• lidé MeSH
• mužská infertilita etiologie   MeSH
• poruchy sexuálního vývoje s karyotypem 46, XY diagnóza   genetika   patologie   patofyziologie   MeSH
• profáze meiózy I MeSH
• segregace chromozomů * MeSH
• spermie patologie   MeSH
• srovnávací genomová hybridizace MeSH
• translokace genetická * MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

Hybridization and introgression can impact the evolution of natural populations. Several wild canid species hybridize in nature, sometimes originating new taxa. However, hybridization with free-ranging dogs is threatening the genetic integrity of grey wolf populations (Canis lupus), or even the survival of endangered species (e.g., the Ethiopian wolf C. simensis). Efficient molecular tools to assess hybridization rates are essential in wolf conservation strategies. We evaluated the power of biparental and uniparental markers (39 autosomal and 4 Y-linked microsatellites, a melanistic deletion at the β-defensin CBD103 gene, the hypervariable domain of the mtDNA control-region) to identify the multilocus admixture patterns in wolf x dog hybrids. We used empirical data from 2 hybrid groups with different histories: 30 presumptive natural hybrids from Italy and 73 Czechoslovakian wolfdogs of known hybrid origin, as well as simulated data. We assessed the efficiency of various marker combinations and reference samples in admixture analyses using 69 dogs of different breeds and 99 wolves from Italy, Balkans and Carpathian Mountains. Results confirmed the occurrence of hybrids in Italy, some of them showing anomalous phenotypic traits and exogenous mtDNA or Y-chromosome introgression. Hybridization was mostly attributable to village dogs and not strictly patrilineal. The melanistic β-defensin deletion was found only in Italian dogs and in putative hybrids. The 24 most divergent microsatellites (largest wolf-dog FST values) were equally or more informative than the entire panel of 39 loci. A smaller panel of 12 microsatellites increased risks to identify false admixed individuals. The frequency of F1 and F2 was lower than backcrosses or introgressed individuals, suggesting hybridization already occurred some generations in the past, during early phases of wolf expansion from their historical core areas. Empirical and simulated data indicated the identification of the past generation backcrosses is always uncertain, and a larger number of ancestry-informative markers is needed.

• MeSH
• beta-defensiny genetika   MeSH
• chromozom Y MeSH
• genetická variace MeSH
• genetické markery * MeSH
• genotyp MeSH
• hybridizace genetická * MeSH
• mikrosatelitní repetice MeSH
• mitochondriální DNA MeSH
• molekulární evoluce MeSH
• multilokusová sekvenční typizace * MeSH
• populační genetika MeSH
• psi MeSH
• shluková analýza MeSH
• vlci MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• psi MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Itálie MeSH