[Genomic instability in atherosclerosis].

A comparative study of the level of genomic instability, parameters of quantitative and structural mutations of chromosomes (aberration, aneuploidy, polyploidy) in lymphocyte cultures from patients with atherosclerosis of age 80 years and older (control group - 30-35 years old) was conducted. The possibility of correction of disturbed genomic indicators by peptide bioregulators - Livagen (Lys-Glu-Asp-Ala) and cobalt ions with separate application or in combination was also studied. Control was lymphocyte culture of two healthy respective age groups. It was also shown that patients with atherosclerosis exhibit high level of genomic instability in all studied parameters, regardless of age, which may suggest that there is marked increase in chromatin condensation in atherosclerosis. It was also shown that Livagen (characterized by modifying influence on chromatin) separately and in combination with cobalt ions, promotes normalization of altered genomic indicators of atherosclerosis in both age groups. The results show that Livagen separately and in combination with cobalt ions has impact on chromatin of patients with atherosclerosis. The identified protective action of Livagen proves its efficacy in prevention of atherosclerosis.

[Genome instability in pulmonary tuberculosis before and after treatment].

Pulmonary tuberculosis is classified as a disease with a genetic predisposition, and therefore, as with other pathologies related to this group of diseases, by pulmonary tuberculosis, special importance is given to finding those markers that enable early identification of risk groups, such as skrinnig in general population and relatives of patients with tuberculosis, which in turn can provide the basis for preventive measures. One of this markers is the level of genome stability. The aim of this study was a comparative evaluation of the functional parameters of the genome variability in patients with sensitive form of pulmonary tuberculosis before and after treatment, and the possibility of its correction with anti-stress peptide bioregulator - epitalon. The studies were conducted using short-term mitoge -stimulated cell cultures of TB patients, before and after treatment. As an indicator of genome stability has been studied the frequency of structural and numerical chromosome aberrations and fragile sites. It is shown, that in intact cultures from patients with pulmonary tuberculosis, before treatment was significantly higher level of frequency of cells with structural chromosome aberrations, that still retained after the treatment. As for epithalon, it appears that was shown a pronounced protective effect after treatment, on the test of chromosome aberrations, by reducing both overall mean frequency aberrant cells and indicators for all individuals. In the study of fragility of chromosomes in patients with primary tuberculosis was found, that in intact cultures, the proportion of cells with chromosomal fragile sites was higher than in control group of healthy individuals, befor and after treatment. High frequency of chromosome fragility persisted by treatment with peptide bioregulator in both cases - before and after treatment. It is suggested that the identified patterns can be correlated with a high incidence of re- TB.

[Deheterochromatinization of the chromatin in old age induced by oligopeptide bioregulator (Lys-Glu-Asp-Pro)].

In this work is presented the data on the variability of the functional characteristics of the chromosomes in the cells exposed by oligopeptide bioregulator - Prostamax from old individuals (75-86 years). Evaluated: the frequency of sister chromatid exchanges (SCE); Ag-positive NORs (in associations and nonassociations), as well as the variability of the structural C-pericentromeric heterochromatin. Prostamax changed the chromosomal parameters: 1) increased the frequency of SCE to 12,0±0,28 exchange in per cell (in intact cells - 5,9±0,2); 2) increased the frequency of Ag-positive NORs to 2.5 per cell (in intact cells - 0.95) 3) reduced in the frequency of large segments of the options from the pericentromeric heterochromatin for the 1st and 9th chromosomes. Comparison of the results indicates the ability of Prostamax to decondensation, deheterchromatinization the chromatin during aging, and thus release by heterochromatinization repressed genes. On the other hand, the data obtained in this work suggest that the basis for the protective action of Prostamax its modifying effect on chromatin.

Effect of the peptide bronchogen (Ala-Asp-Glu-Leu) on DNA thermostability.

Thermodynamic parameters of DNA melting in the presence of a peptide bronchogen in various concentrations were estimated on a differential scanning microcalorimeter. Bronchogen was shown to serve as a DNA-stabilizing agent. Bronchogen increased the melting temperature of DNA from calf thymus and mouse liver by 3.1°C in a narrow range of r (molar ratio of bronchogen/DNA b.p., 0.01-0.055). A further increase in r was not accompanied by changes in the melting temperature. The complex melting enthalpy (ΔH(melt)) remained unchanged in this range of r (0.01-1.0). ΔH(melt) for DNA from the thymus and mouse liver was 11.4 and 12.7 cal/g, respectively. Our results indicate that bronchogen is not an adenine-thymine-specific or guanine-cytosine-specific ligand. The type of binding is considered as strong and occasional. The binding occurs with both strands of DNA (mainly with nitrogen bases).

[Variability of C-structural heterochromatin in cells derived from the patients with cardiomiopathy and from their relatives].

Variability of C-structural heterochromatin segments in chromosome 1,9 and 16 has been studied in lymphocyte cultures of peripheral blood taken from the patients with the hypertrophy (HC) and dilatate (DC) forms of cardiomiopathy and their 1st degree relatives (32 individuals, in total). 10 healthy individuals composed the control group. C-segments were sorted according to Patil and Lubs: a<0.5 x 16p; b>0.5-1 x 16p; c>1.5 x 16p; d>1.5-2 x 16p; e>2 x 16p. The total amount of C-heterochromatin in all the studied chromosomes was tended to increase for DC patients and the relatives of the patients with the two forms of cardiomiopathy. Individual specificity within the group was found when the c-variants were assessed in chromosomes. In particular, the results obtained in cells of HC patients and their relatives did not differ from the control values, while the distribution pattern of C-segments within the group of DC patients and their relatives underwent changing. The elevated induces of pericentromeric inversion were found in all the patients with both forms of the disease and their relatives indicating C-structural heterochromatin polymorphism in the tested individuals.

[The effect of heavy metal ions and peptide bioregulators on the expression of chromosome fragile sites in the individuals of different age groups and breast cancer patients].

Expression rates of chromosome fragile sites in peripheral blood lymphocytes have been studied in clinically healthy individuals of different age groups (20-38 yrs and 75-86 yrs) and breast cancer patients (8 cases). In individuals with a normal check-up of different age groups the heavy metal (nickel, zinc and cobalt) ions were also examined on their influence on the expression of the fragile sites and the peptide bioregulators (Livagen and Epithalon) were tested on their ability to correct the pattern of expression. Short-term lymphocyte cultures were used as tested material. The analysis showed that the chromosomes of people from young and old age groups differ from each other by the expression pattern of fragile sites - the chromosomes of young individuals were found to be more active by spontaneous formation of fragile sites. They were also sensitive to their induction by heavy metals. Both tested bioregulators lessen heavy metals effect that was statistically reliable only for the young people group. As for the patients with breast cancer general elevated fragility of chromosomes and specific distribution of the fragile sites along the chromosomes were revealed.

[Variability of radiation-induced adaptive response in old age individuals and their correction by Peptide bioregulator -Livagen].

Effect of aging on adaptive response of cellular systems to low (stimulated) dozes of gamma-rays (0, 2 and 0, 5 Gy) and to disturbing dozes of radiation (1 and 2 Gy) has been investigated. PHA-stimulated cells were from 72-86 year-old individuals; control - 30-40 year-old individuals. The potentialities of induction of adaptive response in cells exposed to previously irradiated by stimulating dozes of gamma-rays with subsequent damaging effect of copper chloride (10(-3)M) has been investigated. The correcting activity of the peptide bioregulator Livagen was tested. The investigation showed that cells from aged individuals maintained radiation adaptive feature. Preliminary exposure to radiation caused stimulation of adaptive response in copper-treated cells. Corrective activity of Livagen was observed.

[Chromosome instability in patients with different forms of cardiomyopathy and their relatives].

Comparative studies on spontaneous and induced by two agents (mytomycin C and Nickel chloride) chromosome instability have been conducted in cultured lymphocytes derived from patients with hypertrophic and dilated forms of cardiomyopathy (HCMP and DCMP) and their relatives compiling a high-risk group to develop mentioned pathologies. In patients with HCMP and DCMP, as well as in their relatives higher levels of spontaneous chromosome instability were found in comparison with the control group (comprising healthy individuals without cardiomyopathic anamnesis). Almost all the affected patients and their relatives revealed tendency to increased aneuploidy and significant elevation of polyploidy when compared with control values. It was demonstrated that nickel chloride-induced chromosome disorders registered more frequently in lymphocytes of the patients and their relatives than in cells of control subjects, but mytomycin C did not have such an affect. We assume to explain the similarity of studied parameters in patients with cardiomyopathy and their relatives as a consequence of identical rearrangements in genome functioning. The revealed analogy should be interpreted as a pre-illness condition for the individuals included in the risk-group.

Effects of short peptides on lymphocyte chromatin in senile subjects.

Effects of synthetic short peptides (Vilon, Epithalon, Livagen, Prostamax, and Cortagen) on activity of ribosome genes, parameters of common heterochromatin melting, polymorphism of structural heterochromatin (C segments) of chromosomes 1, 9, and 16, and variability of facultative heterochromatin were studied in leukocytes of subjects aged 75-88 years. All the studied peptides induced activation of ribosome genes, decondensation of densely packed chromatin fibrils, and release of genes repressed as a result of age-specific condensation of the cellular euchromatin regions (deheterochromatinization of facultative chromatin). Treatment with Epithalon, Livagen, and Prostamax led to decondensation of chromosome 1 pericentromeric structural chromatin, while Epithalon and Livagen treatment led to changes in chromosome 9 as well. Hence, short peptides activate heterochromatin and heterochromatinized regions of cell chromosomes in senile subjects.

Effects of Livagen peptide on chromatin activation in lymphocytes from old people.

We studied the effects of the synthetic peptide Livagen on activity of ribosomal genes, denaturation parameters of heterochromatin, polymorphism of structural C-heterochromatin, and variability of facultative heterochromatin in lymphocytes from old people. Livagen induced activation of ribosomal genes, decondensation of pericentromeric structural heterochromatin, and release of genes repressed due to age-related condensation of euchromatic regions in chromosomes. Our results indicate that Livagen causes de-heterochromatinization (activation) of chromatin, which is realized via modification of heterochromatin and heterochromatinized regions in chromosomes from old people.

[Human chromosome functional characteristics and aging]. / Funktsional'nye osobennosti khromosom cheloveka i starenie.

The MUTATION level (chromosome aberrations covering telomere regions), MODIFICATIONS of CHROMOSOME STRUCTURE (level of condensed chromatin identified by the methods of electron microscopy and differential scanning microcalorimetry; level of C-banding constitutive heterochromatin; transcriptional activity of DNA-dependent RNA polymerase; Ag-positive NORs and associations of acrocentric chromosomes) and REPARATION (intensity of unscheduled DNA synthesis and the frequency of sister chromatid exchanges) have been studied in lymphocyte cultures from individuals at the age of 70-114 to reveal the chromosome functional organization at late stages of ontogenesis and to find explanations of some senile pathologies. The analysis of obtained results showed: 1) chromosome progressive heterochromatinization (condensation of eu- and heterochromatin regions) occurs at aging; 2) decrease of repair processes and increase in frequency of chromosome aberrations in aging are secondary to the progressive heterochromatinization. Chromosome heterochromatinization is a key factor of aging; 3) chromosome heterochromatinization may be the reason for some senile pathologies; 4) chromosome heterochromatinization is an area where one should seek the ways for prolonging the lifespan.

[Structural mutations of chromosome mutations induced by heavy metal salts during aging in vivo and in vitro]. / Izuchenie strukturnykh mutatsii khromosom indutsirovannykh soliami tiazhelykh metallov pri starenii in vivo i in vitro.

The cytogenetic effect of inorganic copper (CuSO4) and cadmium (CdCl2) salts was studied in cells of long-term (144-hour) human cultures (in vitro model of aging) and in elderly individuals (80-93 years old). Copper sulfate increased the incidence of aberrant cells both in elderly individuals (14.25 +/- 1.74%) and during in vitro aging (12.20 +/- 1.62%) (3.94 +/- 1.96% and 5.25 +/- 1.10% in the controls, respectively), whereas treatment with cadmium chloride did not induce any changes in the background index. Differences in the effect of the studied salts may be due to their different effects on chromatin modification.

[Characteristics of the cis- and trans-positions of chromatids of human acrocentric chromosomes in extreme old age]. / Kharakteristika tsis- i trans-polozhenii assotsiiruiushchikh khromatid akrotsentricheskikh khromosom cheloveka v glubokoi starosti.

Cis- and trans-positions of chromatid associations of human acrocentric chromosomes were examined at extreme old age. Lymphocyte cultures were prepared by the usual method, from peripheral blood of 9 subjects aged 80-90 years (analysis of 179 metaphases) and 7 subjects aged 20-48 years (analysis of 124 metaphases). The functional difference between stalks of the sister chromatids was found. In the subjects at the age 80-90 years satellite stalks of chromatids-1 (in all DNA strands thymidine was substituted by 5-BrdU) of the D chromosome in cis-position are included into associations with lower frequency, as compared with the satellite stalks of chromatids-2 (thymidine+ is only substituted by 5-BrdU in a half DNA strands of the chromosome). This apparently reflects variability of regulation of functional activity of satellite stalks of sister chromatids.

Heterochromatinization as a key factor in aging.

An analysis of observations on modifications during aging of the characteristics of chromosomes during the mitotic cycle of their mutations and unscheduled aging leads to the conclusion that enhanced heterochromatinization during aging prevents the action of reparative enzymes, and results in increased numbers of cells with chromosomal aberrations.

The activity of nucleolar organizer regions of human chromosomes in extreme old age.

Ag-positive nucleolar organizer regions (NORs) and associations of silver-stained acrocentric chromosomes were studied in 400 metaphases from 20 male and female subjects aged 20-50 and 80 or over. The number of Ag-positive NORs and the frequency of chromosome associations were found to be decreased significantly at 80 and thereafter in comparison with middle age. The use of G stain revealed a significant age-dependent decline in the numbers of argent-affine 13th and 21st chromosomes and the frequency of associations formed by the 13th and 22nd chromosomes. These phenomena seem to be determined by heterochromatinization of satellite stalks of the 13th, 21st and 22nd chromosomes during aging.

[Nucleolus organizer region activity of human chromosomes in relation to age]. / Aktivnost' iadryshkoobrazuiushchikh raionov khromosom cheloveka v zavisimosti ot vozrasta.

Ag-stained acrocentric chromosomes were cytogenetically studied in 20 individuals aged 20-50, 80 and in older ones. It is revealed that the number of Ag-positive nucleolus regions and association frequency of Ag-stained acrocentric chromosomes statistically decreased in the individuals of old age as compared with the middle-aged persons. This is supposed to be conditioned by heterochromatinization of the satellite stalks of the 13th, 21st and 22 d chromosomes in the process of ageing.

[Sister chromatid exchanges in human lymphocytes in extreme old age]. / Sestrinskie khromatidnye obmeny v limfotsitakh cheloveka v glubokoi starosti.

After the treatment with 5-bromodeoxyuridine (BrdU), the frequency of sister chromatid exchanges (SCE) in the lymphocyte culture cells of peripheric blood tends to be lower in very aged persons, from 80 to 88 years (7.69 +/- 0.34), than in middle aged ones (8.40 +/- 0.33). The reliable decrease in SCE level was assessed in chromosomes A1 and C from the old group persons as compared with the appropriate values in the middle age group. The level of SCE falls in telomeric and centromere regions of chromosomes A1 and A2.

[Human acrocentric chromosomal associations in old age]. / Assotsiatsii akrotsentricheskikh khromosom cheloveka v starcheskom vozraste.

Associations of satellite chromosome fibres of acrocentric chromosomes of the DD, GG and DG types were studied in persons aged from 80 to 114 years. It was found that the value of P (probability of joining of satellite fibres of two chromosomes in a group) is considerably less than the corresponding value for persons aged from 20 to 48 years. It is supposed that the decrease of the frequency of association at senile age is caused by heterochromatization of satellite fibres in the 13th and 14 chromosome pairs.

[Weakening of the ultraviolet ray-induced unscheduled DNA synthesis in human lymphocytes in extreme old age]. / Oslablenie indutsirovannogo ul'trafioletovymi luchami vneplanovogo sinteza DNK v limfotsitakh cheloveka v glubokoi starosti.

The intensity of unscheduled DNA synthesis was studied in UV-irradiated (10--15 J/m2) peripheral blood lymphocytes of 80--90 years old persons. In these extreme old age persons, reparative DNA synthesis was found sufficiently reduced in comparison with that in middle aged (20--43 years old) ones. The role of DNA repair processes in ageing is under discussion.