An analytical method for studying DNA degradation by electrophoresis after cell lysis and visualization of DNA fragments with fluorescent dye, comet assay, was used to evaluate the viability of the endothelial layer of human arterial grafts with the aim of identifying the procedure that will least damage the tissue before cryopreservation. Four groups of samples were studied: cryopreserved arterial grafts that were thawed in two different ways, slowly lasting 2 hours or rapidly for approx. 7 minutes. Arterial grafts that were collected as part of multiorgan procurement with minimal warm ischemia time. Cadaveric grafts were taken as part of the autopsy, so they have a more extended period of warm ischemia. The HeadDNA (%) parameter and others commonly used parameters like TailDNA (%). TailMoment, TailLength, OliveMoment, TailMoment to characterize the comet were used to assess viability in this study. The ratio of non-decayed to decayed nuclei was determined from the values found. This ratio for cadaveric grafts was 0.63, for slowly thawed cryopreserved grafts 2.9, for rapidly thawed cryopreserved grafts 1.9, and for multi-organ procurement grafts 0.68. The results of the study confirmed the assumption that the allografts obtained from cadaveric donors are the least suitable. On the other hand, grafts obtained from multiorgan donors are better in terms of viability monitored by comet assay. Keywords: Arterial grafts, Cryopreservation, Cadaveric, Multiorgan procurement, Viability, Comet assay.
- MeSH
- arterie transplantace MeSH
- kometový test * MeSH
- kryoprezervace * MeSH
- lidé MeSH
- mrtvola MeSH
- přežívání štěpu fyziologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
The myocardial extracellular matrix plays an important role in maintaining the structural and functional integrity of the heart and is centrally involved in post-myocardial infarction repair processes. We analysed some genetic and proteomic aspects that could play an important role in the development of myocardial infarction. Matrix metalloproteinases are enzymes that contribute strongly to the degradation of extracellular matrix components. In this study the serological levels of MMP-2 and MMP-9 were investigated using immunological testing in 34 patients with myocardial infarction and 34 matched control subjects. The serum levels of MMPs were determined by ELISA. Changes in serum levels were characterized within 24 h and after 6 months post myocardial infarction. Significantly higher levels of MMP-2 (299.47 ± 117.61 ng/ml) and MMP-9 (93.56 ± 53.74 ng/ml) were determined in patients with myocardial infarction compared to the controls, in both cases P < 0.001. MMP-9 levels decreased significantly in the 6 months after cardiac event, whereas the levels of MMP-2 were almost equal to the post-infarction ones. While comparing the results from four patients that died of cardiovascular cause within 6 months we found significantly higher MMP-2 (435.00 ± 55.83 ng/ml, P = 0.003) and MMP-9 (166.25 ± 41.07 ng/ml, P = 0.018) values. Microarray analysis was used to determine the gene expression of selected genes for MMPs and their regulators from peripheral blood. The selected genes did not show satisfactory results that could have a potential implication for diagnostics of tissue degeneration.
- MeSH
- infarkt myokardu krev enzymologie genetika MeSH
- lidé středního věku MeSH
- lidé MeSH
- matrixová metaloproteinasa 2 krev MeSH
- matrixová metaloproteinasa 9 krev MeSH
- regulace genové exprese MeSH
- senioři MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- matrixová metaloproteinasa 2 MeSH
- matrixová metaloproteinasa 9 MeSH
- MMP2 protein, human MeSH Prohlížeč
- MMP9 protein, human MeSH Prohlížeč
Non-Governmental Organization Archaia (http://www.archaia.cz) carried out the rescue archaeological research at Knezeves near Prague in 1998. Most of dating objects in Knezeves come from the period of Late and Final Bronze Age. The approximately 3,000 years old set, which included 11 human remains from three settlement features, was collected for the study. First, gender was determined according to anthropological characteristics. Ancient DNA from bones was extracted by the phenol-chloroform procedure and N-phenacetylthiazolum bromide reagent. Polymerase chain reaction amplification of AMEL XY, part of amelogenin gene, with subsequent polyacrylamide gel electrophoresis and Short Tandem Repeats analysis followed. DNA profiles of skeletal remains were obtained by the fragmentation analysis of autosomal short tandem repeat markers. Genetic profiles showed us whether individuals from Knezeves were in mutual relationship (parent - descendant). The congruence of results in sex determination supported reliability of genetic methods, which are suitable for sex determination of fragmental and subadult skeletal remains.
- MeSH
- archeologie metody MeSH
- dějiny starověku MeSH
- DNA analýza MeSH
- kosti a kostní tkáň chemie MeSH
- lidé MeSH
- mikrosatelitní repetice MeSH
- polymerázová řetězová reakce metody MeSH
- sekvence nukleotidů MeSH
- sekvenční analýza DNA metody MeSH
- Check Tag
- dějiny starověku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- historické články MeSH
- publikace stažené z tisku MeSH
- Názvy látek
- DNA MeSH
Diabetes mellitus type 2 represents a heterogenous disease characterized by impaired glucose homeostasis. The disorder clusters in families suggesting genetic disposition, however the mechanism underlying is unknown. Many studies show more frequent maternal transmission of diabetes in the families. One of huge range of explanation is exclusively maternal transmission of mitochondria. Mitochondria are power organelles which produce ATP molecules by oxidation-reduction reactions via the respiratory chain. They contain their own genome which codes subunits of the respiratory chain and proteosynthetic apparatus for proteins encoded by this genome. Pathogenic mutations of mitochondrial DNA can affect the activity of the respiratory chain and result in various phenotypes. Mitochondrial diabetes is commonly associated with neuromuscular disorders and often presents with nonautoimmune beta cell failure. Although mitochondrial mutations are associated with diabetes, their low frequency does not explain reported more frequent maternal transmission of diabetes mellitus type 2.
- MeSH
- diabetes mellitus 2. typu genetika MeSH
- lidé MeSH
- mitochondriální DNA genetika MeSH
- mitochondriální nemoci diagnóza genetika MeSH
- mutace MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
- Názvy látek
- mitochondriální DNA MeSH
In the recent past, application of DNA genotyping techniques has enabled researchers to more accurately test relationships between dominance rank (DR), mating success (MS) and reproductive success (RS). Paternity studies often reveal that reproductive outcome does not always correlate with male DR and/or MS and thus open room for discussion and interpretation of alternative reproductive tactics of both sexes. In this study, we analysed male DR, MS and RS in a group of bonobos at Twycross Zoo (UK). Genetic relationships were determined using 8 tetrameric microsatellite loci. Despite clear and asymmetric dominance relationships, analysed using normalised David's scores based on a dyadic index of dominance among the group's 3 mature males, we found that the most dominant male did not sire the most offspring. In fact, both infants conceived during the observation period were found to be sired by the lower-ranking males. Although the alpha male had almost exclusive mating access to one of the females during the time she was showing a maximal anogenital swelling, her infant was sired by the lowest-ranking male who mostly mated with her when outside the maximal swelling period. This result suggests that either sperm competition operates and/or ovulation is decoupled from the phase of maximal anogenital swelling which could allow greater female choice.
- MeSH
- DNA fingerprinting MeSH
- DNA analýza MeSH
- fertilita fyziologie MeSH
- mikrosatelitní repetice MeSH
- Pan paniscus genetika fyziologie psychologie MeSH
- paternita MeSH
- rozmnožování fyziologie MeSH
- sexuální chování zvířat fyziologie MeSH
- sociální chování MeSH
- sociální dominance * MeSH
- zvířata v ZOO MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Názvy látek
- DNA MeSH
DNA profiling--inclusive sex determination-- with microsatellite markers is currently a commonly used genetic method of studying humans. An efficient technique of producing the genetic data is amplification of multiple microsatellites in a single PCR reaction. Here we introduce a novel PCR-multiplex system for analysis of four polymorphic Y-STRs. Specifically, these are DYS449, DYS456, DYS458, and DYS464. These loci were chosen because of their reported high diversity in Euroamerican population (10), as well as their absence in the commercial analytical kits at the time of beginning of this study. Our objective was to design this PCR-multiplex for use of fragmentation analysis by electrophoresing samples on a capillary semi-automated genetic analyzer applying only one fluorescent dye. The PCR system we propose, may be notably used in fields such as forensic and human population genetics.
The AB0 blood group system typing remains one of the basic laboratory tasks in a forensic practice. However, problems arise when the analysed samples are seriously degraded. The DNA analysis promised to solve this but an unexpected complication was encountered. For the AB0 blood group system typing a Polymerase Chain Reaction method was used to amplify glycosyltransferase gene, when DNA had been isolated from artificially created blood stains, followed by their subsequent artificial thermal degradation. In the B0 genotype an aberrant genotype was suprisingly found and its structure was confirmed by sequencing. This meant that a newly formed B00 (not the original BO) genotype was present. Such a finding, to our best knowledge, had not been observed yet and we were unable to find any references in the professional literature. The explanation of this result thus remains unclear.
- MeSH
- ABO systém krevních skupin genetika MeSH
- alely MeSH
- fenotyp MeSH
- genotyp MeSH
- lidé MeSH
- polymerázová řetězová reakce MeSH
- sekvenční analýza DNA MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
- Názvy látek
- ABO systém krevních skupin MeSH
The AB0 blood group system typing remains one of the basic laboratory tasks in a forensic practice. However, problems arise when the analysed samples are seriously degraded. We took blood samples from six volunteers (three men, three women) and made blood stains on pieces of sterile cotton cloth. Blood stains were incubated at three different temperatures (22 degrees C, 37 degrees C, 56 degrees C) for various periods of time (1 day, 1 week, 14 days, 1 month, 3 months, 6 months, 1 year). For blood stains degraded at 22 degrees C we also analysed the samples after 3.5 hours of incubation. Moreover, we tried to determine the AB0 blood group system after thermal degradation at high temperature, accurately at 200 degrees C for 10 min. For the AB0 blood group system typing a Polymerase Chain Reaction method was used to amplify glycosyltransferase gene, when DNA had been isolated from artificially created blood stains, followed by their subsequent artificial thermal degradation. For serological AB0 typing the mixed agglutination and the Therkelsen method were used. The DNA analysis seemed to solve problems with seriously degraded blood stains but we found out that classical serological methods were even better in some cases.
- MeSH
- ABO systém krevních skupin * genetika MeSH
- aglutinační testy MeSH
- krevní skvrny * MeSH
- lidé MeSH
- polymerázová řetězová reakce MeSH
- sekvenční analýza DNA * MeSH
- soudní lékařství MeSH
- určování krevní skupiny a křížové zkoušky * MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
- Názvy látek
- ABO systém krevních skupin * MeSH
During a check-up of a driver by a police patrol in the Czech Republic by means of a breath analyzer of Dräger Co. an alcohol blood level of 0.50 g/kg was assessed. Later the driver reported that before the test he used at 30- and 5-minute intervals the drug Stopangin spray. In the Institute of Forensic Medicine the authors made an experiment which revealed that 22 minutes after administration of the drug the apparatus gave a negative result. Agreement with these conclusions was expressed also by a representative of Dräger Co. and the results were published as an expert opinion with a recommended procedure for the police of the Czech Republic in the South Moravian region.
- MeSH
- aerosoly MeSH
- dechové testy přístrojové vybavení MeSH
- ethanol krev terapeutické užití MeSH
- faryngitida farmakoterapie MeSH
- lidé MeSH
- otrava alkoholem krev diagnóza MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
- kazuistiky MeSH
- Názvy látek
- aerosoly MeSH
- ethanol MeSH
UNLABELLED: A missence mutation Trp64Arg in the beta3-adrenergic receptor gene is associated with obesity, insulin resistance, a lower metabolic rate and the earlier onset of NIDDM but the published results are controversial. We investigated the effect of this mutation on insulin resistance (euglycemic hyperinsulinemic clamp), on fat mass and fat distribution (anthropometry, bioimpedance, CT) and resting metabolic rate (indirect calorimetry), lipid spectrum and other metabolic disturbances in Czech juveniles recruited from juvenile hypertensives (H, n = 68) and controls (C, n = 81). The frequency of this mutation (determined by digestion of 210 bp PCR product with MvaI) was double in H than in C (14.7%, vs. 7.4%) and the carriers of Arg64 allele had sig. higher fasting glucose (H: p = 0.002. C: p = 0.025). Four Trp64/Arg64 and six Trp64/Trp64 men (age 23 +/- 4.2, vs. 22.5 +/- 1.9 y, BMI 26 +/- 5.5, vs. 22.9 +/- 5.1 kg/m2) took part in a detailed pilot study. But no signif. differences (Horn's method) in fasting glucose (4.6 +/- 0.6, vs. 4.9 +/- 0.4 mmol/l), in parameters of insulin resistance (M-value150-180 min. 9.1 +/- 1.1, vs. 8.9 +/- 1.5 mg glucose/kg.min(-1)), resting metabolic rate/lean body mass (RMR/kg LBM: 78.6 +/- 4.6, vs. 85.6 +/- 23.2 kJ/kg), lipid spectrum and other screened parameters were found. The lowest resting metabolic rate (RMR/kg LBM 55.4; 62.6 kJ/kg) was found in brothers (both C, Trp64/Trp64) who highly differ in body constitution (BMI 19.0 resp. 32.4 kg/m2). We suppose that in this case the energy metabolism is probably determined by other genetic loci and does not correlate with body fat mass. CONCLUSION: Our pilot study does not confirm the influence of Trp64Arg mutation in heterozygous carriers on insulin resistance, energy metabolism and lipid spectrum.
- MeSH
- beta-3-adrenergní receptory genetika MeSH
- dospělí MeSH
- energetický metabolismus genetika MeSH
- heterozygot MeSH
- index tělesné hmotnosti * MeSH
- inzulinová rezistence genetika MeSH
- lidé MeSH
- lipidy krev MeSH
- metabolický syndrom genetika MeSH
- mutace * MeSH
- substituce aminokyselin MeSH
- tuková tkáň patologie MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- beta-3-adrenergní receptory MeSH
- lipidy MeSH
