The Electrical Conductivity of Bone Marrow Cells Suspension Associated with Proliferative Activity and Depends on the Donor's Age and the Functional Activity of the Liver.

Copper ions play various roles in mammalian cells. Some investigations indicated that copper correlates with liver fibrosis and changes in bone marrow cells. Here, we explored the role of bone marrow cell parameters in copper liver fibrosis development and possible underlying mechanisms. The age-related features of biochemical indicators, bone marrow cell parameters, and electrical conductivity of 3- and 20-month-old male Wistar rats treated with copper II sulfate pentahydrate (CuSO4 × 5H2O) have been investigated. Based on the obtained earlier experimentally data in our laboratory, dose of 1 mg/100 g (0.58 mg/g of liver) was used. Induced liver fibrosis caused a change in the number and ratio of morphotypes of bone marrow cells, calcium and copper content, redox-system activity. These parameters depended of animals' age and presence of liver fibrosis. We also demonstrated that electrical conductivity can be used as integral parameter. In conclusion, the "adaptive response" modifies the emerging reactions of the organism to new external factors.

[Liver-induced fibrosis in young and old animals is accompanied by age-dependent changes in bone marrow cells.]

The hypothesis of dependence of the functional characterizations of bone marrow cells (BMC) - proliferation rate, direction proliferation, etc. - not only by the age of animals, but also on features of BMC microenvironment was verified. Two methods of changing the microenvironment were used. There were in vivo (induction of liver fibrosis in young and old animals) and in vitro (transfer of young and old animals BMC obtained in intact animals and animals with fibrosis into the same standard culture system). CuSO4-induced liver fibrosis and CCl4-induced liver fibrosis had a different effect on the ratio of cell types in the bone marrow in young and old animals. Thus, in young animals, regardless of the type of liver fibrosis inducer, the relative number of morphologically identifiable cell types decreased. This was accompanied by an increase in the number of identified cell types against the background of CuSO4-induced liver fibrosis and did not change against the background of CCl4-induced liver fibrosis in old animals. The proliferative activity of BMC isolated from old animals and transferred to an in vitro culture was superior to that of young animals. This is due to the large number of lymphocytes in the bone marrow of old animals by 167% and the specific composition and characteristics of the BMC microenvironment in old animals.

[Young and old animals use different strategies for forming an immune response to infectious agents (Pseudomonas aeruginosa и Escherichia coli).]

Features of the immune response to injection of suspension of bacteria (Pseudomonas aeruginosa and Escherichia coli) in young (3 months) and old (20 months) Wistar rats at 3, 5 and 7 days after infection were investigated. The dynamics of the content of circulating immune complexes (CEC), complement C3 fragments, and the activity of oxygen-dependent and oxygen-independent phagocytosis were determined. It was shown that infection was accompanied by a multiple increase in the content of circulating immune complexes in young and, especially, in old animals. It remained at a high level from the 3d to the 7th day of the development of pathology in young animals, while they content decreased to 7 days in old animals. On the background of the development of the infectious process, the content of complement C3 fragments decreased in young animals, and it increased in old animals, in the case of infection with P. aeruginosa and E. coli. The activity of oxygen-dependent and oxygen-independent phagocytosis decreased in different ways in young and old animals on the background of infections, i.e. the ratio of the forms of phagocytosis in old and young animals was different. Old animals were not inferior to the young in terms of the ability of the immune system to respond to the presence of an infectious agent. So they used a different strategy for forming the immune response.

[Donor age affects on the «behavior¼ and the sensibility bone marrow cells in on copper ion of the primary culture].

The changes of bone marrow cells (BMC) number in the primary culture from 0 to 96 hours, the pattern (the distribution of cells) of cells morphotypes and «lifespan¼ (the time of cell life after isolation) of myelocytes, metamyelocytes, band and segmented neutrophils, isolated of the young (3 months) and old (20months) animals, were investigated. The number of the BMC obtained from intact old animals increased faster in primary culture, than from young animals. The Cu induced fibrosis had different influence on the rate of BMC culture growth of old and young animals. The adding of 4 mM and 8 mM CuSO4x5H2O in the BMC culture of young and old animals resulted in a dose-dependent inhibition of growth rate of young animal cells. If copper ions were added into the culture of BMC of old animals, the decreased of the BMC number was described less than for cells of young animals. The adding of 8 mM CuSO4x5H2O inhibited proliferation less, than the adding of 4 mM CuSO4x5H2O. The Cu-induced liver fibrosis had accelerated the BMC rate death of both old and young animals. However, this effect was more pronounced in young animals. It is suggested, that during the ontogenesis the BMC undergo such epigenetic changes, which change functional properties.

Young and old animals use various strategies of metabolic adaptation to Cu-induced liver fibrosis.

The role of the prooxidant-antioxidant and immune systems in formation of age-related adaptive responses to chronic effects of toxic substances (copper ions) was studied. In this purpose in young and old animals by multiple sequential administration of copper sulfate was induced liver fibrosis. Using this model the characteristics of prooxidant-antioxidant and immune systems, and physiological characteristics of were assessed. The Cu-induced liver fibrosis was accompanied by the oxidative stress and a significant reduction in cell immune activity. In the case of elimination of oxidative stress by introducing exogenous antioxidants (mix factor) in animals with fibrosis was recovered efficiency, body weight, the content of red blood cells. In old animals the restoring of the changed parameters of cellular immunity was more effective than in the young. Young and old animals used different metabolic strategies of adaptation to toxic environmental factors.

Thermogenesis and longevity in mammals. Thyroxin model of accelerated aging.

Development of experimental models of life span regulation is an important goal of modern gerontology. We proposed a thyroxin model of accelerated aging. Male Wistar rats at the age of 17 months received thyroxin in drinking water at a concentration of 6 mg/L for 2 months as a model of induced hyperthyroidism (IH). Administration of thyroxin resulted in a decrease in life span and a 2°C increase in body temperature that was accompanied by a 2 fold increase in thyroxin level and a 40% increase in triiodothyronine in blood serum. Induced hyperthyroidism can be used as a model of accelerated aging. We also found that thyroxin administration acts as uncoupler of oxidative phosphorylation as treatment was accompanied by an increase in the generation of superoxide radicals by 50%. Antioxidant enzyme activity remained unchanged (glutathione peroxidase, glutathione reductase mitochondrial) or was reduced (glutathione-S-transferase by 1.7 times) as compared with the control. The activity of glucose-6-transferase was increased by 2.8 times as compared with control, and malate dehydrogenase activity in liver increased by 6.8 times. Induced hyperthyroidism in rats resulted in distinct epigenotype which was accompanied by a decrease in life span.

[Cyclic feeding regime--a new model for experimental gerontology].

The influence of specific cyclic feeding regime (CFR) providing 30% loss of body weight for 14 days and 14 days recovery period followed by the standard feeding diet in the course of 30 days, on rat life span was investigated. It has been shown that young and old animals differed in intensity loss and recovery of body weight after three cycles CFR. The mass of liver changes cyclically as well as body weight. It was found that the content of glucose and protein in serum was cyclically changed only in the first cycle of CFR; during the second cycle of the same feeding regime a completely different trend of these parameters was observed. Such nature of the dynamics on the same type of repetitive exposure indicates change in adaptation strategies. This indicates the usage of alternative metabolic pathways that provide survival in extreme conditions. Each subsequent cycle of weight loss-restoration differed from the previous one and had a pronounced age-related character. Animals that have successfully passed three consecutive CFR beginning from 19-months of age had longer life span as compared with the control animals group. CFR can be used as a new model in gerontology providing increased life when transferred on a diet not only young but also old animals.

[Metabolic memory enhances hormesis effect to the copper ions in age-depended manner].

The ability of young and old rats to manifest the hormesis effect to lethal doses of copper sulphate and the ability to save the induced "adaptive" pattern of redistribution of copper ions after the transfer of animals in the standard conditions is the mechanism of metabolic memory. It was found that pretreatment of animals with low-dose (1 mg per 100 g body mass, i.e. 33% of the lethal dose) of copper sulfate induced the formation of their resistance to lethal doses (3 mg per 100 g), so the hormesis effect was manifested. Hormesis effect depended on the number of pre injections of small doses of copper sulphate in an S-shaped manner. The protective effect increased after 1 to 3 of preliminary injections of copper sulfate, and after four or more injections the hormesis effect decreased. It is shown that the cardinal role in intracellular pattern of copper ion redistribution play heat-stable copper binding proteins 12 kDa cytosolic proteins. The formed "adaptive" pattern of intracellular distribution of the copper ions may be reproduced, after at least, one month. The prolonged hormesis effect can be attributed to the forming metabolic memory. The intracellular distribution pattern of the copper ions was age-dependent. Age-related differences were found in hormesis effect induced by copper ions, which results in increased binding capacity of copper binding proteins in old animals, with a higher content of copper ions in the mitochondria and microsomes as compared to young animals.

[Is it possible to "cancel" aging process of cell cultures under optimal conditions for cultivation?].

The characteristics of the cells epigenotypes Dunaliella viridis Teod. in the process of chronological and replicative aging were investigated. By 40th day of accumulative cultivation (which coincided with the stationary growth phase) DNA content in the cells of Dunaliella viridis increased 2 times, triacylglycerides 3 times, beta-carotene and carbonyl proteins 2 times, RNA content decreased in comparison with cells in exponential growth phase, i. e., the 40th day of growth of culture forms the age-related epigenotype. 4 received subcultures were being transplanted during 2 years in mid-logarithmic growth phase (subculture-10), early stationary phase of growth (subculture-20), in the mid-stationary growth phase (subculture-30), and late stationary growth phase (subculture-40). It is shown that epigenotype of subculture-10 remained unchanged over 2 years of cultivation, i. e., it does not manifest replicative aging. At the same time, the subculture-20, although long enough (at least 40 passages), maintained epigenotype characteristic of young cultures, and showed age-related changes. Pronounced age-dependent changes of epigenotype in the course of cultivation were identified for subculture-30, and subculture-40 was characterized by unstable epigenotype. Thus, cultivation conditions determine the intensity of replicative aging in Dunaliella viridis.

Therapeutic efficacy of silymarin and naringenin in reducing arsenic-induced hepatic damage in young rats.

We investigated the effects of silymarin and naringenin in counteracting arsenic-induced hepatic oxidative stress post exposure. Male wistar rats were chronically exposed to sodium arsenite for eight months followed by oral treatment with silymarin and naringenin (50 mg/kg each) for 15 consecutive days to evaluate hepatic damage and antioxidant potential. Our results demonstrate a significant decrease in hepatic GSH levels, SOD and catalase activities and an increase in GST and TBARS levels after arsenic administration. Silymarin or naringenin administration increased GSH levels and was beneficial in the recovery of altered SOD and catalase activity besides significantly reducing blood and tissue arsenic concentration. Our results point to the antioxidant potential of these flavonoids, which might be of benefit in the clinical recovery of subject exposed to arsenic. These flavonoids can be incorporated into the diet or co-supplemented during chelation treatment, and thus may afford a protective effect against arsenite-induced cytotoxicity.

[Imprinting effect on the pattern of intracellular distribution of copper ions in the liver following repeated administration of copper sulfate].

Fractions of copper-binding protein (CBP) specifically bound copper ions were extracted from the rat liver cell cytosole. These fractions of 10-14 kDa proteins are involved in specific pattern of intracellular distribution of copper ions. The imprinting effect of specific pattern of copper ions intracellular distribution has been found. The effect was detected 30 days after sequented injections of copper sulfate into the body. It was shown, that after primary injection of copper the ability of CBP to bind copper ions could increase tenfold, regardless of schemes of copper sulfate injection.

[Supposed role of "metabolic memory" in formation of response reaction to stress-factors in young and adult organisms].

The influence of the combined long-lasted influences of sulfur sulfate and diet restriction in young (3 month age) and adult (21 month age) Vistar rats on activity of glucose-6-phospatase, alaninaminotranspherase (ALT), aspartataminotranspherase (AST), and on phosphorilating activity of liver mitochondria was studied to investigate the role of metabolic memory on the peculiarities of response reaction. The young animals not differed from adult ones in the possibility of inducing activity of glucose-6-phospatase, ALT, and on phosphorilating activity after the influence of sulfur sulfate and diet restriction. The age-related differences in glucose-6-phospatase and transpherases and phosphorilating activity existing in control disappeared after the long-lasted action of sulfur sulfate and diet restriction. The answer reaction in enzyme activity to stress factors applied many times depends upon the metabolic memory formed in the process of adaptation, and the age of animals have no influence on it. In some relation the ontogenesis may be considered as a result of adaptation genesis. The metabolic memory can change the answer of the system to the stress influence. There are three types of modification of the answer to stress factors: the answer remains unchanged (metabolic memory), "paradox answer" formation, and super activation of the metabolic system.

Calorie restricted diet induces alternative pathways of lipid metabolism for support of proliferative processes in regenerating liver.

This study deals with quality and quantity lipid composition of blood serum and liver, DNA synthesis activity (incorporation of H3-thymidine) in liver in 24 h after partial hepatectomy (PH) in 22-month-old Wistar rats, maintained for 21 months on calorie restricted diet (CRD) and on standard diet ad libitum (SD). The contain of lipids in blood serum and activity of RA-label incorporation (14C-Na-acetate) in serum lipids in 24 h after PH were the same in CRD-fed and SD-fed animals. Quantitative and qualitative composition of lipids in microsomes of regenerating liver also was the same for both groups of rats. In regenerating liver of CRD-fed animals lipid contain in cytosol was 1,8-fold more, but pool of lipid droplets (LD) was 1.5-fold less than in regenerating liver of SD-fed animals. Activity of RA-label incorporation in lipids of microsomes, cytosol and LD pool of regenerating liver of CRD-fed animals was significantly higher, than in SD-fed ones. Activity of RA-label incorporation in lipid fractions and its distribution among cytosol lipids and LD pool lipids differed significantly between SD- and CRD-fed animals. Activity of DNA synthesis in regenerating liver of 22-month-old animals on CRD and SD was the same. It is supposed that calorie restriction induces alternative pathways of lipid metabolism to support proliferation processes in liver after PH.

[The growth constraining diet causes various strategy of adaptation in young and adult animals].

The glucose-6-phosphatase activity of microsomes and phosphorylative mitochondrial activity of liver cells, alanine- and aspartate aminotransferase activities in plasma of blood of young (3 month) and adult (21 month) male rats kept under three various diets resulting in a various degree of a growth inhibition and losses of weight of a body have been studied. There were selected diets giving to animals the food full-valued by composition but limited by quantity. Experiment proceeded 50 days. It was found out various metabolic strategy of adaptation in young and adult animals in reply to this influence. Experimentally this phenomenon revealed itself in various correlations between researched metabolic parameters at young and adult rats. Activity of researched enzymes in adult animals changed in a greater degree in comparison with a base level than at young ones. Researchers suppose that the revealed distinctions of metabolic strategy of adaptation in young and adult animals can be attributed to their generated epigenotypes.

[Does the liver ability to regeneration decrease with age? The dynamics of functional activity of mitochondria in process of the liver regeneration]. / Snizhaetsia li sposobnost' pecheni k regeneratsii s vozrastom? Dinamika funktsional'noi aktivnosti mitokhodrii v protsesse regeneratsii pecheni.

The ability of liver of 1-, 3-, 12- and 24-month rats to mass restoration, mass gain and activity of cell energy system from 1 to 30 days after ectomy of liver medial and left lateral lowels were investigated. It was shown that the old animals did not rebate to young ones in respect of regenerant gain and exceeded them with mitochondria functional activity at the early stages of regeneration (2-24 h after operation). We detected a little lag in old rats (12 and 24 monthes) in comparison with 1-3 ones in rate of liver mass reset to corresponding age groups control level. It may be explained by the difference between young and old animals in strategy of organism homeostasis after ectomy of 2/3 liver mass. The rhythmic character of activities for the most of indices of mitochondria functional activity in liver regenerating up to 24 h with maximum at 2-3 h and 12-18 h after operation was detected. The detected rhythm was typical mostly to mitochondria of the old animals. The high functional activity of mitochondria of the old animals and on the late stages of liver regeneration was shown.

[Metabolism of endogenous aldehydes: participation in oxidative stress-induced lesions and its age aspects]. / Metabolism éndogennykh al'degidov: uchastie v realizatsii povrezhdaiushchego deistviia oksidativnogo stressa i ego vozrastnye aspekty.

This review summarizes the data regarding the synthesis and utilization of endogenous aldehydes and mechanisms of their cytotoxic effect. Peroxidation of polyunsaturated fatty acids is the main source of endogenous aldehydes. There are many different aldehydes generated in the cell. The most abundant is 4-hydroxy-2,3-nonenal synthesized from linoleic acid. Aldehydes may react with proteins and nucleic acids and change their functional properties. Aldehyde utilization mainly occurs in reactions catalysed by aldehyde dehydrogenase, aldehyde reductase and glutathione-S-transferase. The major pathway of their catabolism is accompanied with their conjugation with glutathione. Endogenous aldehyde utilization has its tissue- and age-dependent specificity. The status of aldehyde catabolism can modulate free radical alteration effect on the cell. An adequate stimulation of endogenous aldehyde utilization in the situation of enhancement of free radical generation may promote increasing the cell resistance to oxidative stress injury. Senescence is accompanied by a decrease in endogenous aldehyde utilization intensity in tissues. This could be important in the pathogenesis of age-dependent pathology.

[Effect of combination of high-voltage pulses and other physical factors on the intensity of reproduction of Anabaena flos aquae]. / Vliianie kompleksa vysokovol'tnykh impul'sov i drugikh fizicheskikh faktorov na intensivnost' razmnozheniia Anabaena flos aquae.

It was shown that the influence of high-voltage pulses (intensity of electric field 40-120 kV/cm) on cyanobacteria Anabaena flos aquae can be different and depends on the mode of treatment. It was found that in the range of pulse durations less than 1 microsecond with a pulse repetition rate of up to 500 Hz, and final temperature in the working chamber (60 +/- 1) degree C, a gradual full and irreversible inactivation of cyanobacteria independent of the pulse shape takes place. At the same time, a rapid (but slower than by the action of high-voltage pulses) heating without treatment with high-voltage impulses at the same final temperature leads to an inactivation of 80% of initial cyanobacteria followed by growth of bacteria up to 700% and more.

[Effect of human fetal liver cells on the rate of DNA and nRNA synthesis in regenerating rat liver]. / Vliianie émbrional'nykh kletok pecheni cheloveka na sintez DNK i iaRNK v regeneriruiushchei pecheni krys.

The model of partial hepatectomy was utilized to investigate the influence of human fetal liver cells (FLCs) and of human embryo tissue (PCF) postnuclear cytoplasm fraction injection on the rate of DNA and nRNA synthesis in regenerating rat liver cells. Single infusion of FLCs and PCF into the spleen pulp of rats has been shown to increase the DNA synthesis 24 hours after the operation in 2.5 +/- 0.4 and 3.2 +/- 0.5 times respectively. A group of rats has been identified with no influence of the infusion on the DNA synthesis 24 hours after partial hepatectomy, this process having been even retarding in 48 hours after the operation. Meanwhile the FLCs and PCF infusion enhanced the intensity of nRNA synthesis in 72 hours after the operation in all the animal groups. The effect demonstrated is probably caused by the biologically active substances contained in the fetal tissues.

[A low-calories diet as a model of life span expansion and study of mechanisms of aging]. / Nizkokaloriinaia dieta kak model' uvelicheniia prodolzhitel'nosti zhizni i issledovaniia mekhanizmov stareniia.

The model of experimental augmentation of life expectancy animal with the help of a low-calorie diet designed empirically more 50-[symbol: see text] of years back. The translation animal on such diet is accompanied by development of stress--response and the further adaptation to restricted power supply. As a result of it occurs complex change in all metabolic systems of an organism (energy system of a cell, endocrine, immune system, system of toxicity release, connecting tissue, activity genetic and system which synthesizes an protein). The orientation of these changes provides retardation of development, delay of formation age-dependent of pathologies, and as the consequence, augmentation average life and, that is especially important, maximal life expectancy. The effect of augmentation of life expectancy at use of low-calorie diets depends on quantitative restriction of a calorific within the framework of 40-75%, sex and genotype animal, age animal at the moment of translation on a diet. The proposition expresses, that the prolongation of life at use to a low-calorie diet can be surveyed as specific response of some organisms in a population (not everything, the part animal perishes in a start of experiment) on such stressful action as restriction of nutrition.

[Pharmacokinetics of lipophen--a novel combined phospholipid liver protective agent of natural origin]. / Farmakokinetika lipofena--novogo kombinirovannogo fosfolipidnogo gepatoprotektora prirodnogo proiskhozhdeniia.

Study of the labeled lipophen distribution in organs and tissues of experimental animals showed evidence of a nonlinear drug pharmacokinetics. The maximum amount of labeled lipophen accumulated in liver, spleen, and kidneys, while the drug content in heart, muscles, and brain was very small. The products of lipophen metabolism are excreted with both urine and feces.