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1.
Tihomir Todorov Albena Todorova Bilyana Georgieva Vanyo Mitev 《Molecular biotechnology》2010,45(2):150-154
We report on a unified rapid betaine-based-PCR protocol for amplification of the (CAG)n region in Huntington disease (HD) and the (CGG)n region in Fragile X syndrome (FXS), followed by an electrophoretic separation on automated sequencer for precise determination
of the triplet numbers. The high betaine concentration (2.5 M betaine) permits precise amplification of the CAG and CGG repeats.
Ten HD affected patients and 10 healthy individuals from HD families were re-evaluated. For FXS the CGG region in normal individuals
and premutations of about 100 repeats were precisely amplified by this protocol. Ten unrelated FXS premutation carriers and
24 mentally retarded non-FXS affected boys were re-examined by this method. The results totally coincided with the previous
ones. This protocol is a good choice as a fast screening test. Within 24 h we can have preliminary information on the patient’s
genetic status. Normal individuals, CGG premutation carriers up to 100 repeats, as well as HD patients carrying an expansion
up to 50 CAG repeats can be easily clarified. This accounts for a relatively large proportion (about 90%) of the suspected
HD and FXS patients, referred to our laboratory for genetic analysis. The calculation of the repeat’s number is more accurate
for the correct interpretation of the results, screening tests and genetic counselling. 相似文献
2.
Mild micrococcal nuclease treatment of rat and mouse nuclei and fractionation were based on the method of Tata and Baker. Three chromatin fractions, S, P1, P2, were separated, and for each of these fractions the sensitivity to the DNase 1 action was determined. The relative content in these fractions of non-transcribed DNA sequences was established by hydridization with a mouse satellite DNA, and the relative content of transcribed DNA sequences--by hydridization with DNA synthesised on the total poly (A) mRNA. None of the fractions displayed the properties characteristic of active chromatin. 相似文献
3.
Two pesticides, the fungicide Endodan (ethylene thiuram monosulphide) and the insecticide-acaricide Kilacar (bis(parachlorophenyl)cyclopropyl methanol), produced or used in the neighbouring countries of Bulgaria and Greece were investigated in a coordinated research programme for their genotoxic effects in a variety of test systems. This included the Ames test, Aspergillus nidulans for mitotic segregation, in vitro human lymphocyte cell cultures for SCE and chromosomal aberrations, in vivo bone marrow cells in hamsters and rats and the dominant lethal test in rats. The genotoxicity of Endodan was found to range from negative to slightly positive in different test systems. At concentrations of 7.5 and 12.0 micrograms/plate together with S9 mix it induced base-pair substitutions in the TA100 strain of Salmonella typhimurium at a rather low level. At a dose of 93 mg/kg b.w. it also caused chromosomal aberrations in acutely treated hamster bone marrow cells. A significant increase of SCE was also found in human lymphocyte cultures at a concentration of 20.0 micrograms/ml. Endodan was found to be negative in A. nidulans for somatic segregation, lymphocyte cultures for chromosomal aberrations and mitotic activity and in rats for dominant lethals and chromosomal aberrations. Kilacar was found to be a weak mutagen in the TA97 strain of S. typhimurium at concentrations of 2.5 and 5.0 micrograms/plate together with S9 mix. At concentrations of 1.0, 1.5 and 2 micrograms/ml Kilacar increased the number of mitotic segregants in A. nidulans by 160%, 220% and 156% respectively over the control. In Syrian hamster bone marrow cells after acute administration at concentrations of 0, 40, 80 and 160 mg/kg, the MI was 5.50, 4.30, 3.10 and 1.30 respectively, and an increase in chromosomal aberrations of about 300% over the control was observed with a concentration of 80 mg/kg. In human lymphocytes no significant changes were observed in either MI or SCE. In the dominant lethal test after chronic treatment of male rats at doses of 5.1, 10.2 and 102.0 mg/kg b.w. no significant mutagenic effect was found although a decrease was shown in the percentage of females with implants mated with treated males in the first week. 相似文献
4.
5.
mtDNA diversity in rhesus monkeys reveals overestimates of divergence time and paraphyly with neighboring species 总被引:4,自引:0,他引:4
Reconstructions of the human-African great ape phylogeny by using
mitochondrial DNA (mtDNA) have been subject to considerable debate. One
confounding factor may be the lack of data on intraspecific variation. To
test this hypothesis, we examined the effect of intraspecific mtDNA
diversity on the phylogenetic reconstruction of another Plio- Pleistocene
radiation of higher primates, the fascicularis group of macaque (Macaca)
monkey species. Fifteen endonucleases were used to identify 10 haplotypes
of 40-47 restriction sites in M. mulatta, which were compared with similar
data for the other members of this species group. Interpopulational,
intraspecific mtDNA diversity was large (0.5%- 4.5%), and estimates of
divergence time and branching order incorporating this variation were
substantially different from those based on single representatives of each
species. We conclude that intraspecific mtDNA diversity is substantial in
at least some primate species. Consequently, without prior information on
the extent of genetic diversity within a particular species, intraspecific
variation must be assessed and accounted for when reconstructing primate
phylogenies. Further, we question the reliability of hominoid mtDNA
phylogenies, based as they are on one or a few representatives of each
species, in an already depauperate superfamily of primates.
相似文献
6.
Effects of the whole extract and the chromatographic fractions of the pig placenta on lymphocyte proliferation and humoral immune response 总被引:4,自引:0,他引:4
The immunoregulatory properties of pig fetal placenta extracts (PE) from 1 st, 2nd and 3rd month of pregnancy and five fractions (F1 to F5), isolated on Sephadex G-200 and additionally characterized by fast performance liquid chromatography, FPLC (Superose 12 HR) were studied in order to clarify the local immune regulation in diffuse epitheliochorial placentation. The obtained substances were added at 6.25 to 100 microg in cultures of Concanavalin A-stimulated mouse splenocytes and Phytohemagglutinin-stimulated pig and human PBL to monitor their influence on [(3)H]Thimidine uptake in proliferating lymphocytes. Their effects on the number of plaque-forming cells in spleen cell suspensions from mice treated ip simultaneously with sheep red blood cells and with 100 microg protein of PE, respectively, of each fraction were also investigated: PE and F1 had no effect while F4 and F5 suppressed the mitogen-induced lymphocyte proliferation in all studied species. F2 and F3 stimulated mouse and pig lymphocyte proliferation. The effects were dose-dependent and the suppression was not due to cytotoxic effects. The FPLC data allowed the suggestion that 110 kD protein(s) were involved in stimulation and 7 kD substance(s) - in suppression of cell proliferation. The PE from the 3 studied periods as well as the 5 fractions increased significantly the primary humoral immune response against T-dependent antigen. The results revealed that trophoblast of epitheliochorial placenta produces simultaneously immuno-stimulatory and -suppressive factors acting across the species barrier. Their presence at the feto-maternal interface may contribute to the regulation of local immune reactions and survival of the allogenic fetuses despite the morphological specificities of this type of placentation. 相似文献
7.
G Brosch E I Georgieva G López-Rodas H Lindner P Loidl 《The Journal of biological chemistry》1992,267(29):20561-20564
Mono Q ion exchange high performance liquid chromatography (HPLC) reveals that the main histone deacetylase activity (HD1) of germinating Zea mays embryos consists of multiple enzyme forms. Chromatography of HD1 after treatment with alkaline phosphatase yields two distinct histone deacetylase forms (HD1-A, HD1-B). The same is true for chromatography after phosphatase treatment of a total cell extract. One of these enzyme forms (HD1-A) is subject to phosphorylation, which causes a change in the substrate specificity of the enzyme, as shown with HPLC-purified individual core histone species; the substrate specificity for H2A increases more than 2-fold after phosphorylation, whereas the specificity for H3 decreases to about 60%. The total histone deacetylase activity is quantitatively released from isolated nuclei after extraction with moderate ionic strength buffers; no significant residual enzyme activity could be detected in the nuclear matrix. 相似文献
8.
Dessislava Staneva Ekaterina Peycheva Milena Georgieva Toni Efremov George Miloshev 《Antonie van Leeuwenhoek》2013,103(1):143-152
Kluyveromyces lactis, also known as dairy yeast, has numerous applications in scientific research and practice. It has been approved as a GRAS (Generally Recognized As Safe) organism, a probiotic, a biotechnological producer of important enzymes at industrial scale and a bioremediator of waste water from the dairy industry. Despite these important practical applications the sensitivity of this organism to genotoxic substances has not yet been assessed. In order to evaluate the response of K. lactis cells to genotoxic agents we have applied several compounds with well-known cyto- and genotoxic activity. The method of comet assay (CA) widely used for the assessment of DNA damages is presented here with new special modifications appropriate for K. lactis cells. The comparison of the response of K. lactis to genotoxins with that of Saccharomyces cerevisiae showed that both yeasts, although considered close relatives, exhibit species-specific sensitivity toward the genotoxins examined. 相似文献
9.
Vesela Yancheva Elenka Georgieva Stela Stoyanova Iliana Velcheva Dóra Somogyi Krisztián Nyeste László Antal 《Acta zoologica》2020,101(4):431-446
The present study aimed to investigate for the first time the health status of the Caucasian dwarf goby Knipowitschia caucasica (Berg, 1916, Fishes of freshwaters of Russian Empire, p. 563, Moscow, Russia: Dep. Zemledeliya) (Teleostei: Gobiidae) from an anthropogenically loaded site in Hungary using histopathological analyses on multiple tissues. For that purpose, fish were collected from the public beach at Tiszafüred near the River Tisza. Gills, liver and kidney were subjected to histopathological analyses, and the results showed different alterations in each organ, which also differed in their extent and severity. In addition, we also found lesions in the reproductive organs of both, male and female fish which, overall, we hypothesized could be due to untreated municipal wastewaters, most likely contaminated with endocrine-disrupting chemicals. The multi-organ histopathological analyses of Caucasian dwarf gobies revealed different lesions, prevalence and severity in each target organ, as follows: liver>gills>kidney>gonad (testes and ovaries). These histopathological lesions can be assessed as good indicators of contamination by endocrine-disrupting chemicals of freshwater ecosystems. 相似文献
10.