Biomedical subjects
V N Kalinin
Publications and source records attributed to V N Kalinin.
[A new point mutation in the mitochondrial gene ND1, detected in a patient with type II diabetes].
A novel mutation in a mitochondrial gene was identified in a patient with type II diabetes mellitus. G-to-A transition was localized at the nt3316 position of gene ND1 and resulted in alanine threonine replacement at position 4 of mitochondrial NAD-H-dehydrogenase.
[Analysis of various polymorphic markers of the CFTR gene in cystic fibrosis patients and healthy donors from the Moscow region].
Allelic frequencies at three polymorphic markers in the CFTR gene were detected on chromosomes derived from cystic fibrosis (CF) patients and healthy donors from Moscow and the Moscow region. These polymorphic markers are tetranucleotide tandem repeats GATT in intron 6B, M470V in exon 10, and T854T in exon 14 (fragment A). Frequencies at allele 1 of the M470V marker, along with allele 2 of GATT and T854T, are two times higher for CF patients without delta F508 mutation than for healthy donors, and there is linkage disequilibrium of these alleles of the polymorphic markers analyzed with the CF gene. Allele 1 of M470V and T854T markers, as well as allele 2 of the GATT marker (six repeats), are absolutely linked to mutation F508 of the CFTR gene. Using the polymorphic markers studied, family analysis of CF was carried out in two families.
[Molecular-genetic analysis of certain mutations of the "cystic fibrosis gene" in Moldavia. Characteristics of molecular markers and their linkage with various mutations].
Sixty-one patients with cystic fibrosis (CF) from Moldova were tested for mutations delta F508, G551D, and R553X. Frequencies of various alleles of the repeated GATT sequence in intron 6B of the CFTR gene, their linkage to other polymorphic markers, and various mutations were determined. The frequency of occurrence of mutation delta F508 was only 25%. An absolute majority of CF patients (80%) had pancreatic insufficiency. Mutations G551D and R553X were not found in our sample. Each of 31 chromosomes with mutation delta F508 carries the 6-GATT allele. Most "non delta F508" (78%) and normal (80%) chromosomes were marked by 7-GATT allele. Twenty-seven delta F508 chromosomes (96.4%) belong to haplotype B6, and only one to D6. Most chromosomes with "non delta F508" mutations are associated with haplotypes D7 (26.3%) and C7 (21%). In addition, a significant portion of chromosomes from this subgroup were associated with haplotypes A7 (23.7%), A6 (10.5%), and C6 (2.7%), which are not yet described for mutant chromosomes. The results obtained demonstrate that CF in Moldova is mainly associated with mutations other than delta F508, G551D, and R553X. Severe forms of the disease, with pancreatic insufficiency, are more frequently caused by these mutations; moreover, our data provides strong evidence about the presence of at least seven additional CF mutations in Moldova, apart from delta F508, G551D, and R553X. Some of these are probably not described.
[Maxaquin and ciprofloxacin in the treatment of complicated and recurrent urinary infections in adults].
Clinical efficacy of maxaquin versus ciprofloxacin against complicated or/and recurrent urinary infections (62 patients primarily with chronic pyelonephritis with nephrolithiasis) proved higher (92.3% vs 80.0%) in equal microbiological activity (73.2% vs. 73.4%) of the drugs. Maxaquin was less active in enhancing the culture resistance. Both quinolones had no negative effects on the studied systems of the body's antiinfectious resistance and promoted partial correction of dysbiotic manifestations on the colon mucosa. Side effects of both drugs occurred with similar frequency and severity, starting earlier in administration of maxaquin. The authors offer to manage urinary infections in adults effectively with maxaquin in a dose 400 mg once a day and ciprofloxacin in a dose 500 mg twice a day by 7-14-day courses.
[The creation of an SV40 virus-based vector as an experimental model for the study of gene expression].
The purpose of the studies is to design the recombinant virus SV 40 where the C-end of the basic structural protein of virus P-1 was replaced by a synthetic sequence of the neuropeptide bradykinin. The recombinant virus SV (SV 40/Brd) was obtained. In this virus 60 n.p. with 3'-end of VP-1 gene was substituted for 36 n.p. synthetic gene of bradykinin without impairing the frame of translation. The biological activity of SV 40 (Brd) was tested on the cultured cells CV-1 permissive for this virus. An immunofluorescence method was used to detect T antigen and to examine the cytopathic action of this recombinant. The gene engineering design does not make the recombinant loose its biological properties typical of wild virus.
Altered ratio of collagen chains in bone of a patient with non-lethal osteogenesis imperfecta.
Bone from a patient with osteogenesis imperfecta contained type III collagen which was absent in control bone. The ratio of alpha 1(I)/alpha 2(I) in type I collagen of patient's bone was increased (2.9 vs. 2.3 +/- 0.2 in controls) and the ratio of dimers beta 11/beta 12/beta 22 was altered due to the increased beta 22 content. No abnormality was observed in collagen from the patient's skin. The altered composition of collagen in bone, but the normal composition in skin suggests that the disease in the patient is due to impaired regulation of the synthesis of collagens in bone, rather than by a mutation in one of the two type I collagen genes. Unlike in skin, all the type III collagen in patient's bone was pepsin-soluble indicating an inability of the bone to incorporate type III collagen into mature highly cross-linked extracellular matrix.
Frequency of the F508 deletion in cystic fibrosis patients from the European part of the USSR.
The frequency of the F508 deletion (delta F508) has been analyzed in 189 cystic fibrosis (CF) patients from the European part of the USSR, viz. 127 northern Slavonians (Leningrad region), 30 southern Slavonians (the Ukraine), 10 central Slavonians (Moscow region), 14 Moldavians (Kishenev region) and 8 Lithuanians (Vilnius region). The distribution of CF+ chromosomes with and without delta F508 varied significantly in the different ethnic groups studied and correlated with the clinical manifestation of CF. The overall frequency of delta F508 in Slavonian patients is equal to 62.5%, approximately 90% of them being heterozygous or homozygous for this mutation. The frequency of the deletion among 99 Slavonian patients with severe disease manifestation (pancreatic insufficiency, PI) is equal to 67.5%, only 12 patients having pancreatic sufficiency (PS, 17.5%). The highest value of delta F508 (77.4%) is registered in PI/CF patients of the southern Slavonian group; it is much less frequent (about 57%) in relevant groups of Slavonians from the northern and central parts of the country. Unusually low frequencies (24% and 26%) of delta F508 are detected in a few samples of Lithuanian and Moldavian CF patients, respectively. All delta F508+CF-chromosomes of Slavonian origin are associated with haplotypes 2.2.2. defined by the restriction fragment length polymorphism sites KM19/PstI, CS.7/Hin6I and MP6d-9/MspI, although a high proportion (about 25%) of unknown mutations is associated with the same haplotype. Haplotype B (allele 1XV2c/TaqI; allele 2 KM19/PstI) accounts for 91% of delta F508+CF chromosomes. Our data are consistent with the hypothesis of a single origin and subsequent diffusion of this major CF mutation; however, its interpopulational dissemination in Eastern Europe does not follow the suggested south-east to north-west gradient in Western Europe. The significance of these data for prenatal diagnosis and carrier screening of CF mutations is briefly discussed.
Restriction fragment length polymorphisms of three collagen genes in Russians and Buryats.
The restriction fragment length polymorphisms (RFLPs) of 3 collagen genes have been studied in two USSR populations. Four high-frequency alleles, also reported in other populations, were observed. The frequencies of alleles for the polymorphic sites MspI and EcoRI of the COL1A2 gene, HindIII of the COL2A1 gene and EcoRI of the COL3A1 gene were found to be 0.836, 0.398, 0.447 and 0.786, respectively, in Russians from Moscow (Caucasians), and 0.883, 0.519, 0.648 and 0.809, respectively, in Buryats from the Baikal region (Mongoloids). The RFLPs showed a fair agreement with the Hardy-Weinberg equilibrium. The frequencies were compared with those of other populations.
A crude lysate of cells immobilized on solid support can serve as a matrix for enzymatic DNA amplification.
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Preliminary results on the frequency of the delta F508 mutation in cystic fibrosis patients from the USSR.
The frequency of the delta F508 mutation in a sample of 29 cystic fibrosis (CF) families from the USSR is equal to 44.8%, in agreement with previously reported haplotype data. If confirmed on a larger sample of CF patients from the USSR, this result might be indicative of the presence of a larger heterogeneity of CF mutations in the Soviet population than in other European populations.
[The restriction polymorphism of the gene locus for human growth hormones in different populations of the USSR].
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Modified method for peptide mapping of collagen chains using cyanogen bromide-cleavage of protein within polyacrylamide gels.
A highly efficient method for cyanogen bromide (CNBr)-mapping of collagen peptides is described. This method was developed based on polypeptide cleavage by CNBr within gel slices according to Barsh et al. [1981) Collagen Relat. Res. 1, 543-548). The proposed method has the following advantages: (i) Analysis of both radiolabeled and unlabeled collagens is possible; (ii) CNBr-cleavage of polypeptides is performed in gel pieces which contain individual bands; (iii) The peptide losses are minimized, offering a more complete analysis of collagens including the low molecular weight CNBr-peptides.
The absence of type II collagen and changes in proteoglycan structure of hyaline cartilage in a case of Langer-Saldino achondrogenesis.
Structural analysis of hyaline cartilage extracellular matrix components from the ribs and knee joint of a stillborn female with type II achondrogenesis was carried out. The absence of type II collagen, a decrease in the amount of proteoglycans (PG), and structural changes in PG, namely, increased electrophoretic mobility of PG, lower relative content of chondroitin 4-sulfate (Ch4-S), lower molecular weight and decreased total chondroitin sulfate (ChS) sulfation, were detected. Increased amounts of type I and type III collagens, atypical for hyaline cartilage, were revealed. Among the link proteins (LPs), a large protein with a mol. wt. of 48 kDa was predominant. Molecular and cellular mechanisms of the pathogenesis of achondrogenesis ("chondrogenesis imperfecta") are discussed. The data obtained suggest that the primary defect in type II achondrogenesis involves ChS or type II collagen synthesis.
[Isolation of high molecular weight eukaryotic DNA with the use of potassium acetate].
The simple technique for isolation of high-molecular eucaryotic DNA has been proposed. It includes cell or nuclei lysates by sodium dodecylsulfate in the presence of pronase, proteins precipitation by potassium acetate, DNA precipitation by ethanol. The DNA isolated by this technique is easily cleaved by restriction endonucleases and can be used for analysis of the unique genes by blot-hybridization. The yield of DNA is similar or somewhat higher than that in case of using the standard methods including phenol extraction or phenol-chloroform extraction.
[Optimization of the method of cyanogen bromide mapping of polypeptides separated by polyacrylamide gel electrophoresis].
Two highly efficient methods of CNBr-peptide mapping of polypeptides divided by polyacrylamide gel electrophoresis are described. The first is elaborated on the basis of peptide mapping of collagen proposed by G. Barsh et al. The following three modifications diminish wasting the material essential for the method. 1. CNBr treatment takes place in the absence of CNBr solution outside the gel, excluding the peptides elution from the gel fragments in the process of mapping. 2. After CNBr treatment the solution of CNBr is substituted by the samples buffer before electrophoresis by means of drying and subsequent addition of minimal volumes of the buffer. The latter procedures substitute the gel washing out by the buffer solution. 3. The step of washing the gel fragments by the 70% strong solution of formic acid before CNBr treatment is excluded. The second method of CNBr-peptide mapping is notable for extracting peptides from the gel fragments in the process of CNBr-treatment and permits obtaining of the high quality peptide electrophoregrams.
[Isolation of a restriction endonuclease with HindIII-specificity from Bordetella bronchiseptica].
Site-specific restriction endonuclease BbrI has been found in bacteriophage resistant strain B. bronchioseptica 4994. The technique was elaborated for purification of BbrI to the stage free of nuclease and phosphatase contamination. The yield of purified enzyme is 6000-20 000 units per 10 g of biomass. BbrI recognises and cleaves the same DNA sequence as HindIII with the formation of four-nucleotide cohesive ends. The simplicity of cultivation, security for human, presence of the single restriction endonuclease and the high level of its production make B. bronchioseptica 4994 a promising producer of BbrI restriction endonuclease, isoshizomeric to HindIII, for use in experimental practice in industry.
[Effect of deletions in the ori-Tc region of plasmid pBR322 on its compatibility with replicon pMB1].
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