[Epidemiology and risk factors of chronic renal diseases: a regional level of the problem].

AIM: Assessment of chronic renal disease (CRD) prevalence and morbidity rate and approaches to early CRD in one of the regions of the RF (Tyva Republic). MATERIAL AND METHODS: A population study in the Tyva Republic performed from 01.07.2003 to 30.06.2004 included patients with glomerular filtration rate (GFR) < 30 ml/min, nephrosclerosis signs at autopsy, terminal GFR < 30 ml/min, on replacement renal therapy (RRT). CRD prevalence and morbidity were estimated (stage IV-V). Examination of 374 Tyva citizens was made for estimation of early CRD and risk factors of developing CRD (the sectional study). The participants of the sectional study were examined clinically and biochemically with measurement of albuminuria, calculation of urine albumin/creatinine (ACR) and study of some molecular-genetic characteristics. RESULTS: Prevalence of CRD stage IV-V in Tyva population was 493 cases per million, prevalence of RRT--126 cases per million. Elevated ACR was found in 4.7% of healthy subjects and 15.9% of hypertensive subjects. Initial lowering of GFR occurred in some healthy subjects and in 1 of 12 hypertensive patients. Significant predictors of albuminuria were serum albumin concentration (p < 0.00001), GFR (p < 0.0002), a male sex (p < 0.004), diabetes mellitus (DM) (p = 0.0057) and left ventricular myocardial mass index (p = 0.0253). GFR depended significantly on age (p < 0.000001), male sex (p < 0.000001), uric acid concentration in the serum (p < 0.000001), presence of DM (p = 0.000019), ACR (p = 0.0059), diastolic pressure (p = 0.0347), triglyceridemia (p = 0.0369). Citizens of Tyva had more frequently than citizens of other regions of Russia IlI-genotype of angiotensin 1-converting enzyme (ACE) (p < 0.0001), T-allele of methylentetrahydrofolatereductase (p < 0.0001), E3-allele of gene of apoprotein E (p < 0.0001). Prevalence of aa, ab, bb genotypes of eNO-synthetase was 82.3%, 15% and 2.7% in the group of Tyva examinees vs 62, 31 and 7% in European Russians (p < 0.01). CONCLUSION: Prevalence of both early and advanced stages of CRD among population of Tyva Republic is rather high. CRD morbidity may depend, besides conventional risk factors, some genetic specific features. Screening studies require continuation for early detection of CRD and timely planning of therapeutic and preventive measures.

[Dendrimers based lysine and their "starburst" polymeric derivatives: prospects of use in compacting DNA and in vitro delivery of genetic constructs]. / Dendrimery na osnove lizina i ikh "zvezdoobraznye" polimernye proizvodnye: vozmozhnost' ispol'zovaniia dlia kompaktizatsii DNK i dostavke ékspressiruiushchikh geneticheskikh konstruktsii in vitro.

We attempted to find some compounds for the effective delivery of gene constructs into cells and obtained two trispherical dendrimers on the basis of lysine, (Lys)8-(alpha, epsilon-Lys)4-(alpha, epsilon-Lys)2-(alpha, epsilon-Lys)-Ala-NH2 (D1) and (Lys)8-(alpha, epsilon-Lys)4-(alpha, epsilon-Lys)2-(alpha, epsilon-Lys)-Ala-[Lys(Plm)]2-Ala-NH2 (D2), as well as the starburst polymeric derivatives of D1, (pVIm)8-D1 and (pLys)n-D1, containing poly(N-vinylimidazole) and polylysine chains bound at a single point to the dendrimer amino groups. The conditions of dendrimer-plasmid DNA complex formation were studied. The intracellular localization of these complexes and the expression of gene constructs delivered with their help were analyzed in transfection experiments on the HeLa cell cultures of human epithelial carcinoma and on C2C12 mouse myoblasts. It was found that the chemical structure of dendrimer D1 and its derivatives significantly affected the structure and properties of complex. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 1; see also http://www.maik.ru.

[Expression in vivo of the lacZ gene, delivered to mouse lungs using synthetic microspheres]. / Ekspressiia in vivo gena lacZ, dostavlennogo v legkie myshei pri pomoshchi sinteticheskikh mikrosfer.

A capacity of MF-2 synthetic microspheres to serve as the vehicle for transfer of the marker LacZ gene to mouse lung epithelial cells was studied after a single intranasal administration of the MF-2/gene complex. Two types of plasmids carrying marker gene LacZ were used in the experiments: with cytoplasmic (pCMV-LacZ) and nuclear (pCMV-nlsLacZ) localization of the gene product (beta-galactosidase). As early as 7 days after the complexes MF-2/pCMV-LacZ and MF-2/pCMV-nlsLacZ were administered, specific staining for beta-galactosidase revealed this enzyme activity in the epithelial cells of bronchi, bronchioli, and alveoli. The maximum in vivo of the marker gene in the MF-2/pCMV-LacZ complex was observed at day 14 to 21 after administration and the corresponding gene product was detected during the following two months. The MF-2-mediated gene transfer led to a twofold increase in beta-galactosidase activity relative to the case when the "unbound" pCMV-LacZ plasmid was administered. These results suggest that the synthetic microsphere-mediated transfer of alien genes to the lung of experimental animals is promising. Microspheres may be used in gene therapy for pulmonary affections, in particular cystic fibrosis.

[Population analysis CTG trinucleotide repeats in the myotonin-protein kinase I gene]. / Populiatsionnyi analiz trinukleotidnykh CTG-povtorov v gene miotonicheskoi proteinkinazy I.

CTG trinucleotide repeat length polymorphism within the 3' region of the myotonin protein kinase I (MP-I) gene was examined with the use of the polymerase chain reaction (PCR) technique. A total of 159 DNA samples from healthy donors from five ethnic groups including Russians (n = 33), Azerbaijanians (n = 29), Uzbeks (n = 31), Moldavians (n = 31), and Georgians (n = 35) were analyzed. The number of CTG repeats varied from 5 to 24, and allele distribution was bimodal in all populations; alleles with 6, 7 and 9 repeats were not found. Statistical treatment of the data was performed by the contingency table collapsing procedure. Allele distribution in the populations of Russians, Azerbaijanians, Uzbeks and Georgians appeared to be statistically homogenous and significantly (P < 0.001) different from that in Moldavians as well as in Japanese, African, and Western European populations. High heterozygosity levels of all populations, studied (72-86%) indicate the usefulness of this polymorphism in population and genome fingerprinting studies.

[Correlation of results of immunocytochemical, electrophysiologic, and molecular studies in patients with cystic fibrosis]. / Korreliatsiia rezul'tatov immunotsitokhimicheskikh, élektrofiziologicheskikh i molekuliarnykh issledovanii u bol'nykh mukovistsidozom.

In 13 cystic fibrosis (CF) patients of 5 to 23 years of age with a known mutation spectrum of gene CFTR, sweat chloride values and nasal-potential differences (NPD) were measured and localization characteristics of the protein product of gene CFTR in the cells of nasal epithelium were studied. Sweat Chloride values were normal or boundary (24 to 62 mM/l) in six CF patients. In seven CF patients, these values were significantly above the estimates for the control group. On average, the NPD values were -44.7 +/- 2.2 mV (from -32.5 to -68.9 mV) and -17.2 +/- 1.8 mV (from -6.8 to -30.2 mV) in CF patients and the control group, respectively. Histochemical studies clearly revealed the localization of the CFTR protein on the apical membrane of the nasal epithelium. Depending on the type of mutation, the protein product of gene CFTR was either absent or regularly distributed in the cytoplasm in CF patients; it was not detected in the apical membrane. Thus, NPD measurements and the analysis of the localization of the protein product of gene CFTR in scrapes of nasal epithelium were shown to be additional, highly informative methods of CF diagnostics.

[Expression of marker genes in muscle fibers after transfection in vivo, mediated by lactoferrin]. / Ekspressiia markernykh genov v myshechnykh voloknakh posle transfektsiiin vio, oposredovannoi laktoferrinom.

The capacity of milk iron-transporting human protein lactoferrin (LF) to deliver genetic constructions into cells was studied in an effort to correct hereditary defects. The purified LF and LF conjugates containing either polylysine (C-1) or both polylysine and ficoll (C-2) were bound to plasmid DNA. These complexes were injected into mouse muscles, and the expression of the marker genes was tested immunochemically. Mice were transfected with either pDMD1 plasmid carrying a full-size cDNA for human dystrophin gene or pCMVLacZ plasmid carrying the gene of bacterial beta-galactosidase. The marker gene expression was detected in the muscular fibers. The dystrophin-positive muscular fibers (DPMF) were revealed in mdx mice (a model of Duchenne's dystrophy) in the regions of administration and in muscles of the other limbs. beta-Galactosidase activity was revealed only in the injected muscles. The highest amount of DPMF (9%) was recorded in mice who received the complex of DNA with nonmodified LF. Specific LF-mediated human transfection as a means of stimulating the receptor-mediated endocytosis of genetic constructions and addressed gene transfer to human muscles are discussed.