[Relationships between aging indicators, central hemodynamics and arterial stiffness in men in the European North of Russia.]

With age, the incidence of cardiovascular diseases is steadily increasing worldwide. Living in the harsh climatic and geographical conditions of the Arctic is associated with premature aging, earlier and more frequent development of age-associated diseases. Apparently, these processes are caused by functional changes in large vessels, the state of which has not yet been studied. Aim and methods: to identify correlations of aging and biological age indicators with the parameters of central hemodynamics and arterial stiffness measured using applanation tonometry by the «Sphygmocor¼ device in middle-aged men living in the European North of Russia. For the first time, positive associations of parameters of peripheral and central blood pressure, augmentation index, and negative correlation of the time of return of the reflected wave and pulse pressure amplification with indicators of aging were noted, which indicates an age-related decrease in arterial elasticity. There was also a negative association of the subendocardial viability ratio with the northern experience and biological age. Theoretically considered, the results demonstrate the potential involvement of blood vessels in the pathogenesis of premature aging. The practical significance of the work is related to the need to develop preventive measures aimed at controlling blood pressure, reducing arterial stiffness and cardiovascular risk in middle-aged northerners.

Effect of Liposome-Encapsulated Dextrazide on Local Regulation of Extracellular Matrix Metabolism in Liver and Lungs of Mice with during Chronic BCG-Induced Granulomatosis.

The liposomal form of isonicotinic acid hydrazide conjugate with oxidized dextran (liposomeencapsulated dextrazide, LEDZ) injected intraperitoneally for 3 months to BALB/c mice with chronic BCG-induced granulomatosis (6 month after infection) down-regulated activities of protease (MMP) and antiprotease (TIMP-1, TIMP-2, α2-macroglobulin) components of the regulation system, which indicates a drop of destructive and inflammatory potential of BCGinduced granulomatosis. The persistent enhanced hyaluronidase activity in the liver and its reduced (normalized to control level) activity in the lungs after administration of LEDZ indicates a greater hydrolysis of hyaluronan in the liver than in the lungs. LEDZ-induced changes in MMP/TIMP system in mouse liver and lungs are characterized with relative elevation of protease activity over that of antiprotease one.

Content of the Major Extracellular Matrix Components of the Liver and Lungs in Mice with Chronic BCG-Granulomatosis Treated with Liposome-Encapsulated Dextrazide.

Administration of the liposome-encapsulated dextrazide (LEDZ) to mice 6 months after infection with mycobacterium BCG vaccine for 3 months modulated metabolism of collagens and the intensity of fibrosis of internal organs. In the liver, we observed a redistribution of glycosaminoglycans towards sulfated forms, a decrease in the content of hyaluronan, a change in the ratio of hydroxyproline fractions indicating a decrease in fibrosis via two mechanisms: suppression of synthesis and increased degradation of collagens. In the lungs, administration of LEDZ did not affect the content of sulfated glycosaminoglycans, but significantly reduced the content of hyaluronan, stimulated degradation of collagen, and reduced its synthesis, but these processes were insufficient for significant reduction of fibrotic complications in the lungs. In animals treated with LEDZ, the decrease in collagen synthesis in the liver was 2-fold more pronounced than in the lungs.

[Age-related features of the relationship between the content of vascular endothelial growth factor and indicators of lipid metabolism and extracellular matrix metabolism in men in the European part of the Russian Arctic.]

The content of vascular endothelial growth factor-A (VEGF-A) in blood plasma and its relationship with lipid and extracellular matrix metabolism in working-aged men (19-69 years), living and working in the European part of the Arctic zone of the Russian Federation (Russian Arctic), were studied. No age dependence of the plasma VEGF-A content was found. The correlation analysis, performed in different age groups, revealed significant associations of VEGF-A level with lipid parameters (CS, LDL-C, Apo B, atherogenicity coefficient, Apo B /Apo A1 ratio) and extracellular matrix metabolism (blood TIMP-4, MMP-2, MMP-3, MMP-9, hyaluronan, total and peptide-bound hydroxyproline, glycosaminoglycans). The established correlations indicate the formation of relationships between angiogenesis, atherogenesis and fibrosis at a specific period of life of northerners in the Russian Arctic.

Antifibrotics Effect of Liposome-Encapsulated Composition of Oxidized Dextran and Isonicotinic Acid Hydrazide in Mice with BCG-Induced Granulomatosis Depends on Administration Route.

We studied the response of the extracellular matrix of the lungs and liver in mice with BCGinduced granulomatosis (3 months) after inhalation and intraperitoneal administration of liposome-encapsulated dextrazide (LEDZ): a conjugate of oxidized dextran (40 kDa) and isonicotinic acid hydrazide (INH). LEDZ inhalation proved to be more effective in reducing fibrosis severity, both in the lungs and liver. However, the mechanisms of the antifibrotic effect were different: increased degradation and reduced collagen synthesis in the lungs and reduced collagen synthesis and collagen degradation in the liver. This suggest that drug administration routes and delivery to the target organs are crucially important in the therapy of tuberculosis. The antifibrotic effect depended on LEDZ administration route and was more potent after LEDZ inhalation.

Peculiarities of Collagen Turnover in Aging BALB/c Mice.

Examination of collagen turnover in the liver, lungs, and spleen of BALB/c mice revealed the age-related differences in the levels of hydroxyproline and its fractions. The highest level of total hydroxyproline was observed in the lungs and spleen of young (2-month-old) mice. In the liver, this level attained maximum at the age of 6 months at the expense of elevation of protein-bound hydroxyproline relatively its level in 2-month-old mice. At the age of 12 months, the levels of total hydroxyproline in the liver and spleen were lower than in 6-month-old mice. The decrease in the collagen turnover rate in the liver of 12-month-old mice reflected lower levels of hydroxyproline fractions in comparison with the corresponding values in 6-month-old mice. The rates of collagen turnover in organs differed in mice of different ages: it was maximum in the lungs and spleen of young animals and in the liver of middle-aged mice.

Effect of Liposomal Delivery of Oxidized Dextran with Isonicotinic Acid Hydrazide on Component Profile of Pulmonary Extracellular Matrix in Mice with BCG-Induced Granulomatosis.

Intraperitoneal injections of isonicotinic acid hydrazide (INH), dextrazide (oxidized dextran+INH), or liposomes loaded with dextrazide (INH dose of 14 mg/kg) over 2 months to mice with BCG-induced granulomatosis started from postinfection day 90 induced qualitative and quantitative changes in composition of pulmonary extracellular matrix. Both dextrazide and its liposomal form decreased the levels of sulfated glycosaminoglycans and uronic acids. In contrast to INH, both preparations did not decrease the levels of total glycosaminoglycans, proteins, and galactose. This difference is explained by the fact both free and liposomal dextrazide activated MMP, but did not increase the content of TIMP-1 and TIMP-2, whereas injection of INH was followed by an increase in TIMP-2 content and a decrease in the level of free hydroxyproline, which attested to down-regulation of collagen degradation and maintenance of the conditions for pulmonary fibrosis in mice of this group.

Remodeling of the Extracellular Matrix of the Liver in Mice with BCG-Induced Granulomatosis after Administration of Liposome-Encapsulated Composition of Oxidized Dextran and Isonicotinic Acid Hydrazide.

Remodeling of the extracellular matrix of the liver in mice with BCG-induced granulomatosis after 2-month course of intraperitoneal injections of isonicotinic acid hydrazide, oxidized dextran conjugated with isonicotinic acid hydrazide, dextrazide, and its liposome-encapsulated form manifested in modification of the proteoglycan composition and hydroxyproline fractions associated with changes in hyaluronidase and MMP activities. The antifibrotic effect of dextrazide and its liposome-encapsulated form manifested in reduction of hydroxyproline fractions that reflected the process of collagen synthesis. Administration of isonicotinic acid hydrazide was followed by a decrease in hydroxyproline fractions reflecting collagen synthesis (antifibrotics effect) and degradation (profibrotic effect), which can be explained by its hepatotoxic activity.

Correlations between the Levels of the Main Extracellular Matrix Components in Mouse Liver in Chronic BCG-Induced Granulomatosis.

Correlations between extracellular matrix components in mouse liver revealed the pathogenetically determined dynamic structure of these interrelations (some correlations appeared while others disappeared) at various terms of BCG-induced granulomatosis. The correlations between the studied parameters were strong during the first (postinfection days 3-10) and the second (postinfection days 60-90) periods but became moderate during the third period manifested by chronic inflammation (postinjection day 180). The greatest number of correlations was observed at the beginning of the stabilization period (postinjection day 60). At this period, the content of all structural components of hepatic proteoglycans closely correlated with that of hydroxyproline. These findings attested to systemic changes in metabolism of extracellular matrix components and interrelation between some structural units of proteoglycans in the liver and the components of extracellular matrix in the lungs. The common and typical organ-specific correlations in the liver and lungs suggest the existence of correlations between different organs during progression of BCG-induced granulomatosis.

[Age-related dynamics of the contents of matrix metalloproteinases (MMP-1, -2, -3, -9) and tissue inhibitors of matrix metalloproteinases (TIMP-1, -2, -4) in blood plasma of residents of the European part of the Arctic zone of the Russian Federation.]

Here we present the study of enzymes involved in the regulation of extracellular matrix metabolism. It was the first study of this kind in the European part of the Arctic zone of the Russian Federation (AZRF). The contents of matrix metalloproteinases (MMP-1, -2, -3, -9) and tissue inhibitors of matrix metalloproteinases (TIMP-1, -2, -4) were measured in blood plasma of 91 men in AZRF (67o N) and 14 men in Western Siberia. The contents of MMP-1, MMP-9, TIMP-1 and TIMP-4 in plasma of northerners were higher compared to those of the residents of Western Siberia (55o N). Age-related dynamics of MMP and TIMP contents had a mixed trend in northerners. The maximum contents of MMP-1 and MMP-9 and of MMP-3 were observed in 30-39 and 40-49 years old groups, respectively. The contents of these enzymes tended to decease with age. The maximum contents of TIMP-1 and TIMP-2 and of TIMP-4 were in 50-59 and 60-69 years old groups, respectively, while the minimum contents of these enzymes were in the group of young men up to 29 years old. Hyaluronidase activity was minimal in 30-39 years old group and increased with age up to maximum values at 50-59 years old. Age-related imbalance of MMP/TIMP system (MSE content reduction with age vs TIMP content increase in older age groups) can be considered one of the reasons of the identified age-related increase of interstitial fibrosis and premature aging of the northerners.

Correlations between Extracellular Matrix Components in Mouse Lungs during Chronic BCG-Induced Granulomatosis.

Correlations between extracellular matrix components in mouse lungs were examined during various terms of BCG-induced granulomatosis (on postinfection days 3, 30, 60, 90, and 180). During the development of pathological process, the revealed dynamic interrelations between structural units of proteoglycans and hydroxyproline weakened. Most correlations were observed on postinfection day 180. They reflect the relationships not only between the structural units of proteoglycans but also between collagens, presumably determining the maximum degree of fibrosis at this period. The established correlations characterize the systemic nature of reactions in extracellular matrix and its versatile implications determined by the processes going on in the organs and tissues during the onset and development of generalized pathology.

[Melatonin and man aging in the European part of the Arctic zone of the Russian Federation.]

The analysis of 6-sulfatoxymelatonin (6-SMT) levels in working-age men (aged 19-69) who live in the European part of the Arctic zone of the Russian Federation is presented. Their urine 6-SMT levels were similar to those of comparison group (residents of Western Siberia). The level of 6-SMT decreased with age and was the lowest in 40-49-year group. Age group 60-69 tended to demonstrate increasing 6-SMT levels. Thus, they were insignificantly different (p>0,05) compared to those of people under 29. We showed that alcohol influenced 6-SMT level. The level was lower in alcohol-consuming north men compared to non-consumers. Free testosterone level in plasma of northerners decreased with age. In 60-69-year group, the level was 50% of that in under-29-year group. Compared to controls, we observed the activation of lipid peroxidation that was not compensated by increased activity of glutathione reductase and plasma SH groups. Age-related decrease in the melatonin metabolite, as an endogenous antioxidant, was accompanied by a gradual accumulation of lipid peroxidation products and lower antioxidative protection mediated by SH groups.

[Interrelation between the melatonin level and indicators of aging and fibrosis in men in the European part of the Arctic zone of Russian Federation.]

The results of the correlation analysis between the metabolite melatonin - 6-sulfatoxymelatonin (6-SMT) in the urine and the indicators of aging, interstitial fibrosis, the system of antioxidant protection in men in the Arctic are presented. Negative correlation of medium strength between 6-SMT with calendar age, biological age and molecular marker of aging p16INK4a in leukocytes was noted. A relationship of 6-SMT to fibrosis indices was found. It manifested itself in a negative correlation with sulfated glycosaminoglycans in the urine, with collagen markers: free hydroxyproline, peptide-linked hydroxyproline, and total hydroxyproline. Age-related decrease in the level of 6-SMT may be associated with an age-related increase in interstitial fibrosis in northerners. A positive correlation of medium strength between 6-SMT and hormones: free testosterone level (r=0,53, p=0,0002), total testosterone (r=0,43, p=0,003) and cortisol (r=0,33, p=0,007) was shown. Negative correlations of 6-SMT with the content of ceruloplasmin (r=-0,63, p=0,001) and SCORE scale (r=-0,52, p=0,001) indicate the relationship between melatonin and the antioxidant defense system and the risk of cardiovascular pathology.The results of the correlation analysis between the metabolite melatonin - 6-sulfatoxymelatonin (6-SMT) in the urine and the indicators of aging, interstitial fibrosis, the system of antioxidant protection in men in the Arctic are presented. Negative correlation of medium strength between 6-SMT with calendar age, biological age and molecular marker of aging p16INK4a in leukocytes was noted. A relationship of 6-SMT to fibrosis indices was found. It manifested itself in a negative correlation with sulfated glycosaminoglycans in the urine, with collagen markers: free hydroxyproline, peptide-linked hydroxyproline, and total hydroxyproline. Age-related decrease in the level of 6-SMT may be associated with an age-related increase in interstitial fibrosis in northerners. A positive correlation of medium strength between 6-SMT and hormones: free testosterone level (r=0,53, p=0,0002), total testosterone (r=0,43, p=0,003) and cortisol (r=0,33, p=0,007) was shown. Negative correlations of 6-SMT with the content of ceruloplasmin (r=-0,63, p=0,001) and SCORE scale (r=-0,52, p=0,001) indicate the relationship between melatonin and the antioxidant defense system and the risk of cardiovascular pathology.

[Age-related dynamics of the main extracellular matrix components in residents of the Russian Arctic].

The main extracellular matrix components in Arctic residents were studied. Northerners had increased levels of total glycosaminoglycans, hyaluronan, and collagen IV in plasma and both general and peptide-bound hydroxyproline in urine, which indicates an accelerated metabolism of the main extracellular matrix components compared with comparison group (residents of Siberia). Age-related remodeling of extracellular matrix in northerners manifested in changing ratio and quantity of its main components. Levels of total glycosaminoglycans, hyaluronan, fibronectin, hydroxyproline and its fractions increased with age while the level of collagen IV changed insignificantly. Average positive correlation between extracellular matrix components and biological aging indicators is suggestive of relationship between these two processes: aging - which is accelerated in the Arctic and pathological remodeling of extracellular matrix as it is associated with accelerated aging. Changes in local regulation system including those related to matrix metalloproteinases and their tissue inhibitors may be one of the reasons for pathological remodeling of extracellular matrix.

Altered Liver Proteoglycan/Glycosaminoglycan Structure as a Manifestation of Extracellular Matrix Remodeling upon BCG-induced Granulomatosis in Mice.

Experimental BCG-induced granulomatosis in mice was used to study changes in the dynamics of individual liver proteoglycan components reflecting phasic extracellular matrix remodeling, determined by the host-parasite interaction and associated with granuloma development. In the early BCG-granulomatosis period, the increase in individual proteoglycan components promotes granuloma formation, providing conditions for mycobacteria adhesion to host cells, migration of phagocytic cells from circulation, and cell-cell interaction leading to granuloma development and fibrosis. Later, reduced reserve capacity of the extracellular matrix, development of interstitial fibrosis and granuloma fibrosis can lead to trophic shortage for cells within the granulomas, migration of macrophages out of them, and development of spontaneous necrosis and apoptosis typical of tuberculosis.

Age-Related Changes in the System Metalloproteinases/Tissue Metalloproteinase Inhibitors and Proteoglycan Components in Mouse Organs.

Activity of MMP and content of TIMP and some proteoglycan components were measured in the liver, lungs, and spleen of BALB/c mice aging 2, 6, and 12 months. The increase in the content of proteoglycan components in mouse organs was associated with age-related changes in MMP activity and TIMP-1 and TIMP-2 levels. The ratio between MMP activity and content of TIMP-1 and TIMP-2 differed in the studied organs. The levels of TIMP-1 and TIMP-2 in mouse blood serum and their concentration in studied organs changed asynchronously with age. They are important regulators that determine MMP activity in the analyzed age periods in mice, which can be determined as periods of growth, development, and ageing.

Dynamic Structure of Proteoglycans/Glycosaminoglycans in the Lungs of Mice with Chronic Granulomatous Inflammation.

Structure of proteoglycans in the lungs and total glycosaminoglycan content in blood serum were studied on mouse model of BCG-induced granulomatous inflammation in mice (without destructive processes in the lung parenchyma and granulomas). The maximum level of sulfated glycosaminoglycans in the lungs was detected on postinfection day 30 and was related to their involvement in initiation granulomogenesis and development of granulomas. The maximum level of total glycosaminoglycans in mouse serum on postinfection day 90 coincided with minimum level of sulfated glycosaminoglycans in the lungs. This blood/lungs ratio of glycosaminoglycans can be related to the prevalence of low-molecular-weight hyaluronan fragments promoting inflammation and fibrosis in the lungs observed at the end of the experiment (postinfection day 180).

[Quality of life for men of different ages in the russian European North and its relationship with self-reported health and hormonal status].

Assessment of the quality of life for male inhabitants of the Russian European North demonstrated decreased indices of physical health component (the scale of role functioning determined by physical condition) and mental health component (the scales of social functioning and role functioning determined by emotional state and mental health) compared to the inhabitants of Siberia as a comparison group. Men aged up to 29 had the highest values at all scales. The values decreased with age. The most prominent decrease was observed for the scales of general well-being and pain intensity. There was a moderate decrease for the scales of physical functioning, role functioning determined by emotional state, and role functioning determined by physical condition. Quality of life was closely related to self-reported health of northerners, pathological index, molecular marker of aging, and biological age. Negative correlation between these values implies the deterioration of physical and mental health with biological age. The quality of life for northerners also correlated with hormonal status. There was a relationship between testosterone and three scales of physical health components; between cortisol and one scale of physical component; and between both dehydroepiandrosterone sulfate and insulin and the scales of physical and mental components of health. The effect of various hormones on different age groups of northerners was demonstrated.

Serum, liver, and lung levels of the major extracellular matrix components at the early stage of BCG-induced granulomatosis depending on the infection route.

Experiments on the model of mouse BCG-induced granulomatous showed that the content of glycosaminoglycans and proteoglycans in the extracellular matrix of the liver and lungs are changed at the early stages of inflammation (days 3 and 30 postinfection) before cell destruction in the organs begins. This is related to degradation of extracellular matrix structures. Their high content in the blood and interstitium probably contributes to the formation of granulomas, fibroblast proliferation and organ fibrosis. These processes depend on the infection route that determines different conditions for generalization of the inflammation process. Intravenous method of vaccine injection is preferable to use when designing the experiments simulating tuberculosis granulomatosis, especially for the analysis of its early stages.

[The content of fibronectin in blood serum in patients with myocardium infarction depending on age].

The fibronectin, being one of main components of extracellular matrix of myocardium, is involved into post-infarction repair process. However, the role of fibronectin in this process in patients with myocardium infarction in various periods of their life continues to be a controversial issue. To decide this point the study was organized concerning analysis of content of fibronectin in blood serum in 75 males of three different age groups (prior to 49 years, 50-59 years, 60 -69 years). It is demonstrated that in two groups (prior to 49 years and 50-59 years) the maximal content of fibronectin was marked at second phase regarding to data at first and third phase of reparation process. Whereas in patients aged 60-69 years the maximal content of fibronectin was detected at first phase of reparation process regarding to such in patients aged 50-59 years. The lower content of fibronectin was detected in third phase of reparation process regarding to analogous indicator in other two age groups of patients. These dynamics of content of fibronectin in patients of elder age apparently is an evidence of lower reparation potential of extracellular matrix.