[Stanford Longevity Laws.]

The paper presents the statements and conclusions of the World first unique prospect cohort clinical study on longevity factors - «Genetic Studies of Genius¼, namely, the Stanford study of longevity of «the Termаn's children¼, performed primarily by Dr. Lewis Terman, and later on by Drs. Howard Friedman and Lesli Martin at the Stanford University campus and in City-of-San-Francisco, USA. It was started a century (100) years ago, in 1922. The next statements on longevity were concluded from the study: 1) people («Terman's children¼) with high personal organization live longer; 2) professionally successful people live, on average, ~5 years longer; 3) moderate physical activity and sports contribute to a longer life; 4) activity and high motivation in old age prolong significantly life; 5) people at happy marriages live longer; 6) the sexual satisfaction of husband and wife in a happy marriage predicts a long life for both of spouses; 7) children of divorced parents live, on average, ~5 years less than children in surviving families. It is suggested to call the conclusions of this world's first clinical study on longevity, 1 528 children being involved, as «the Stanford LongevityLaws¼.

[Transdisciplinary approach in free radical theory of ageing. Contribution of Nikolay M. Emanuel and his scientific school to gerontology.]

The work is aimed to review the results of scientific studies of the effect of antioxidants-geroprotectors on the aging of experimental animals and the replicative aging of human diploid cells, carried out in the Department of Kinetics of Chemical and Biological Processes «KHIMBIO¼ of the Institute of Chemical Physics of the USSR Academy of Sciences under the leadership of academician Nikolay Markovich Emanuel in the 1960-1980s after pioneer work by D.Harman. By N.M.Emanuel and colleagues, it was established a previously unknown phenomenon of radical interaction of inhibitors in the oxidation of organic substances, which consists in the regeneration of a more effective inhibitor due to the transfer of a hydrogen atom to its free radical from a molecule of a less effective inhibitor. Antioxidants are polyvalent and can simultaneously affect many stages of aging processes. Data from the N.M.Emanuel scientific school on the increase of the average lifespan of mice by 25,3% and their maximum lifespan by 55,8% using antioxidants, discovered at the Institute of Chemical Physics of the USSR Academy of Sciences as a result of well-founded experimental and theoretical studies, became a powerful argument in favor of the free radical theory of aging in 1970-ties. This was further promoted by approaches based on the theory of reliability, the damage theory, and as well as an approach based on oxidative activation of the Nrf2 signaling pathway, which maintains the «nucleophilic tone¼ of protective oxidoreductases.

[Correlation between Haemostatic and Vegetate Nervous System Parameters under Examination Stress].

The work is aimed to an assessment of the correlation between of autonomic nervous system tone and the level of reactivity of the students' cardiovascular system and hemostasis system under examination stress. It was found that the students under exam stress are characterized with high values of respiration rate and with lower ones of tidal volume. The value of respiratory minute volume decreases in male students due to the bronchoconstrictor effects of stress at the tracheobronchial tree. This finding confirms a statistically significant reduction in the flow of small, medium and large bronchi, indicating an increased parasympathetic nervous system tone. A statistically significant increase in the power of fast waves as a result of determining heart rate variability in students of both hender during the exam also testifies the activation of the vagus nerve and can be used as a marker of exam stress. While exam stress, trombocrite value decreases and only femail students show a statistically significant reduction in the platelet number and increase of their volume. The tendency to increased thrombotic events under exam stress is demonstarated, being reached statistically significant differences in the case of female students--an increase in the initial thrombosis rate. In the case of mail students, an increase of prothrombin time value is testified under exam stress.

[Emotional stress as a clinical model to study the pathogenesis of the initial phase of the general adaptation syndrome].

INTRODUCTION: General adaptation syndrome (GAS), the basis of the development of which is stress phenomenon, is an essential component of the pathogenesis of many diseases and syndromes. However, the patho genesis of GAS hitherto is considered exclusively from the endocrinological viewpoint. This relates primarily to the initial phase of the GAS, a clinical model for the study of which may be psycho-emotional stress (PES), which we studied using three groups of volunteers. METHODS: The first one consists of 25 students who were waiting for unaccustomed physical activity (17 men) and play debut on the stage (8 women). The second group consists of 48 children (2-14 years) who expected for "planned" surgery. The third group of volunteers is made up of 80 students (41 women and 39 men) during the first exam. The concentration of cortisol, endotoxin (ET), the activity of antiendotoxin immunity (AEI) and the haemostatic system parameters were determined in the blood serum of volunteers in various combinations. RESULTS: We found laboratory evidence for PES at 92% of students of the first group, 58% of children of the second one and in 21% of students of the third group of volunteers (mostly women). The concentration of ET increased at 13 (52%) volunteers of the first group with a significant increase of average indicators in the whole group (from 0.84 ± 0.06 to 1.19 ± 0.04 EU/ml). At children of the second group, the average concentration of ET increased even more significantly (from 0.42 ± 0.02 to 1.63 ± 0.11 EU/ml), which was accompanied by the activation of the hemostasis system. A degree of the activation was directly dependent on the level of ET in the general circulation and on an activity of AEI. Examination stress in the third group of volunteers is accompanied by activation of plasma hemostasis (increased initial thrombosis rate and reduced the time it starts, lag-period) in 26% of female students and 15% of male students. CONCLUSION: We suggest that it is possible to use the PES as a clinical model for studying the initial phase of the GAS, examine the role of excess of intestinal ET in the general blood circulation (endotoxin aggression) in the induction of systemic inflammation, which is very likely participated in the initiation of the GAS.

[Fatty acid profile of Pseudomonas aurantiaca DNA-bound lipids according to ESI-LC-MS mass-spectrometry].

Approach to the study of prokaryotic chromatin lipidome has been realized based on analysis of fatty acid profile of DNA-bound lipids using electrospray ionization mass spectrometry ESI-LC-MS. By this method, we found 16 : 0 and 18 : 1 fatty acids, which are contained in the first fraction (weakly bound to DNA), and 14 : 0, 16 : 1 and 18 : 2 fatty acids, which are contained in the second fraction (strongly bound to DNA).

[Stress and allostasis: problems, outlooks and relationships].

Concepts of stress and allostasis are discussed in the paper in their interrelation and interpenetration. General adaptation syndrome is considered in the historical aspect of Claude Bernard's, William Cannon's and Hans Selye's contributions to the field, and from the standpoint of an interdisciplinary approach. Key challenges and contradictions of the stress concept are disclosed and discussed. First, the terminology ones, since this term might be understood as stress response, stressor, state of the organism and even the consequences of stress itself. Second, it is ambivalent and contradictory nature of the stress response. From the perception of stress as a reaction to the strong negative impact of environment, researchers became to divide stress onto two types: distress as a non-specific basis of disease, and eustress as a favorable factor that extends the adaptability of human organism. The third problem is connected to the interdisciplinary nature of the stress response, the effects of which being manifested in, at least, four areas: physiology, behavior, subjective experience, and cognitive functions. Particularily, physiologists are very often ignored of a role of cognitive factors in their stress research. The authors in terms of the allostasis theory, more complex adaptation process than homeostasis, resolve contradictories of stress conception observed. The paper proposes a model of allostatic states to describe distress and eustress phenomena.

[Lipid and fatty acid profiles of Pseudomonas aurantiaca DNA-bound lipids determined by mass spectrometry].

An approach used on investigation of the lipid composition of loosely (fraction 1) and tightly (fraction 2) DNA-bound lipids of Pseudomonas aurantiaca cells by electrospray ionization using mass spectrometry (ESI-LC-MS) was used for determination of the lipidom of a prokaryotic cell. Free fatty acids C16:0, C18:1 (fraction 1), C14:0, C16:0, and C18:2 (fraction 2) were detected. Both fractions of DNA-bound lipids were characterized by the presence of phosphatidylglycerol, phosphatidylserine, and lyso-phosphatidylinositol. The alcohol-soluble fraction 1 could also contain phosphatidylcholine and phosphatidylinositol, while fraction 2 probably contained triacylglycerides. Compared to gas chromatography, ESI-LC-MS provides new possibilities for investigation of the nucleoid lipidome, providing for more detailed investigation of DNA-bound lipids in bacterial cells.

A spectroscopic and surface plasmon resonance study of oleic acid/DNA complexes.

The interaction of synthetic polynucleotide double strands with a natural lipid, oleic acid, was examined in diluted aqueous solutions by circular dichroism spectra, UV-absorption measurements, and surface plasmon resonance biosensor investigations. The investigations were performed with defined double and triple stranded oligo- and polydeoxyribonucleotides. Whereas duplexes are influenced by oleic acid ligandation, which could not be removed by ethanol dialysis procedure, no binding occurs to triple stranded DNA. The spectroscopic results indicate that oleic acid shows molecular recognition to AT b.p. motifs by groove binding. GC tracts - in particular alternating d[G-C] motifs - are strongly influenced by ligand interaction up to a ratio of one molecule per two base pairs. Likewise, the spectroscopic and morphologic changes in the supramolecular association of the complexes after treatment occur even after dialysis procedure. This was monitored with scanning force microscopy (SFM) as well. Additionally, monolayers of biotinylated DNA duplexes were immobilized on a streptavidin sensor-layer for surface plasmon resonance (SPR) observations. Small portions of the ligand were injected in continuous flow. Loosely bound molecules were removed by washing procedure. Injections of sodium hydroxide denature the DNA, releasing the tightly bound effectors. The amount of tightly bound oleic acid molecules was determined at one molecule per 2-3 base pairs. As consequence, a new mechanism of regulation of gene expression at nuclear membrane or by lipids inside DNA double helix has to be discussed.

DNA-bound lipids of normal and tumor cells: retrospective and outlooks for functional genomics.

By very soft phenol method, the high-molecular-mass natural DNA complexes (10(8)-10(9) Da), which contain 1-3% specific lipids, were isolated from different eukaryotic and prokaryotic cells. Two pools of DNA-bound lipids were isolated: loosely bound (extracted with 35% ethanol) and tightly bound lipids (extracted after additional treatment DNAse I). The composition of these two lipid pools of different sources (rat thymus, liver, regenerating liver, loach sperm, pigeon erythrocytes, Zajdel ascites hepatoma, Ehrlich ascites carcinoma, sarcoma 37, Escherichia coli B, T2 phage) was studied. The DNA-bound lipid pools consist of neutral lipids (NL) and phospholipids (PL), moreover NL is always in a few fold more than PL. The composition of these lipid pools of eukaryotes distinguishes between themselves, mainly, by free cholesterol (minor fraction), cardiolipin (major fraction), and by phosphatidylcholine. Only the tightly bound lipid pool was present in T2 phage DNA. The dramatic redistribution effect between all fractions of NL pools (free and ester cholesterol, free fatty acids, diglycerides) was observed in DNA synthesis phase of cell cycle on the background of the unchanged composition of PL pools. Comparative analysis of DNA-bound lipid pools of normal and cancer cells was carried out. The DNA-bound lipid pools of transformed cells significantly differ from the same normal cells both by PL composition (cardiolipin) and by the presence of additional fractions (mono- and triglycerides) as well. The possible functions of DNA-bound lipid pools, especially of cardiolipin and cholesterol at the attachment of DNA loops to the nuclear matrix, DNA replicon organization, replication, and transcription are discussed.

Cationic lipid-DNA complexes-lipoplexes-for gene transfer and therapy.

Cationic lipid-mediated gene transfer and delivery still attract great attention of many gene therapy laboratories. From the point of view of the most important characteristics of lipoplex particles, e.g. its charge and size, we reviewed recent studies available. In general, the paper deals with non-viral systems of gene transfer into eukaryotic cell based on various lipids. Having usually less efficiency in gene transfer, lipid-based gene transfer vehicles (lipoplexes/genosomes) are characterized with certain advantages even over viral ones: they are less toxic and immunogenic, could be targetable and are easy for large-scale production, a size of transferred DNA being quite high. Conditions of DNA condensation during interactions with lipids are described. Results of the studies of mechanism of DNA-lipid complex interactions with the cell membrane and their transport into the nucleus are summarized. Dependence of efficiency of gene transfer on lipoplex structure and physical-chemical properties is reviewed. Advantages and disadvantages of different macromolecule complexes from the point of view of transfection efficiency, possibility of use in vivo, cytotoxicity and targeted gene transfer in certain organs and tissues are also discussed. Results of transfection of different cells using neutral, anion and cation liposomes are reviewed. The conclusion reached was that efficiency and specificity of gene transfer may grow considerably when mixed macromolecule lipid systems including polycations and glycolipids are used.

Specific natural DNA-bound lipids in post-genome era. The lipid conception of chromatin organization.

Two pools of DNA-bound lipids were isolated from DNA supramolecular complex (SC-DNA): loosely bound (extracted with 35% ethanol) and tightly bound lipids (extracted after additional treatment DNase I). The compositions of the two lipid pools from different sources (rat thymus, liver, loach sperm, pigeon erythrocytes, Zajdel ascites hepatoma, Ehrlich ascites carcinoma, sarcoma 37, Escherichia coli B and T2 phage) were studied. The possible functions of DNA-bound lipids, especially of cardiolipin and cholesterol, at the attachment of DNA loops to the nuclear matrix, in DNA replicon organization, replication and transcription are discussed.

Role of lipid membrane-nucleic acid interactions, DNA-membrane contacts and metal (II) cations in origination of initial cells and in evolution of prokaryotes to eukaryotes.

The problems of the origin of primary cells and eukaryotic cells are discussed in terms of possible role of interactions between nucleic acids with lipid membrane according to corresponding original hypothesis. We propose that there are two main hypotheses of the origin of primary cells: (a). RNA appeared before proteins and DNA [Nature 213 (1967) 119]; (b). it is needed for the appearance of a primary cell, the volume closed by the lipid membrane. There was no information about the ways on how RNA appeared inside that volume for saving the reaction products around. Our hypothesis suggests that one of the starting points in the origination of primary cells was the interaction of nucleic acid and lipid membrane bubbles in the presence of metal (II) ions (which existed in high concentrations in prebiotic conditions), and this resulted in the enclosing of the pro-RNAs inside the lipid membrane. This hypothesis is formulated by us on the basis of experimental biochemical and biophysical studies of the DNA/RNA-phospholipid vesicles interactions in the presence of metal ions (II) fulfilled in the Institute of Biomedical Chemistry, RAMS, Moscow and Institute of Biophysics, RAS, Pushchino. Our belief is that DNA-membrane contacts (DNA-MCs) played an important role in the prokaryotes-to-eukaryotes transition. The model of the confluence of four prokaryotic cells may explain the prokaryotes-to-eukaryotes transition by the way of eukaryotic nuclear pore formation from prokaryotic Bayer' contacts. The main requirement for the following fusion of prokaryotic cells must be their mutual orientation. After possible association, the division of the formed cell is begun. The great advantage of the model of four prokaryotic cells is the profit in the metabolism and the possibility of the intensive growth of intercellular membrane structures.

DNA-phospholipid recognition: modulation by metal ion and lipid nature. Complexes structure and stability calculated by molecular mechanics.

The structures and formation energies of nucleic acid-phospholipid complexes both in the absence and in the presence of Mg(2+) ions were calculated taking double-stranded trinucleoside diphosphates NpNpN or heptanucleotides ApAp(NpNpN)pApA, composed of 64 possible combinations of genetic code, and phosphatidylcholine (PC) and sphingomyelin (SM) as model compounds. The dependence of intramolecular interactions on the primary structure of nucleic acid molecules and on the presence of a cationic bridge was revealed. The formation energies and structure of oligonucleotides were found by molecular mechanics calculations with the AMBER force field. The structures of phospholipid and MgCl(2) molecules were calculated by the semiempirical PM3 method, while the energies of phospholipid-oligonucleotide complexes were calculated by the molecular mechanics method. Calculations of complexes were carried out with consideration of solvation effects. Considerable gain in the formation energy of triple complexes is achieved due to the presence of the electroneutral metal bridge. A tendency toward increasing the stability of "triple" PC complexes (but not SM ones), containing guanosine- and cytidine-enriched triplets was revealed. Depending on the structure of NpNpN trinucleotides, the formation energy values of NpNpN-MgCl(2)-PC and ApAp(NpNpN)pApA-MgCl(2)-PC complexes differ by 1.7-2.6 kcal mol(-1), which can be considered as the atomic-scale manifestation of the recognition phenomenon. Presence of metal (II) ion bridge results in a greater stabilization of the phospholipid-nucleic acid complexes for SM in comparison to PC (the total energy difference equals to 4-16 kcal mol(-1)). Depending on the structure of NpNpN trinucleotides, the formation energies of NpNpN-MgCl(2)-SM and ApAp(NpNpN)pApA-MgCl(2)-SM complexes differ by 1.7-2.1 kcal.mol(-1), which is essential at physiological conditions and can also be considered as the recognition effect.

[Metallonucleoliposome complexes as a vehicle for gene delivery to mouse skeletal muscles in vivo]. / Metallonukleoliposomnye kompleksy kak sredstvo dostavki genov v skeletnye myshtsy myshi in vivo.

A simple new method for preparing plasmid DNA and preformed zwitterionic liposome complexes is proposed. The ability of these metallonucleoliposome complexes to serve as a vehicle for gene delivery to mammalian cells in vivo was studied. A high level of expression of the reporter gene introduced was observed in mouse skeletal muscles in vivo.

[Interaction of spin-labeled methacyne analog with butyrylcholinesterase]. / O vzaimodeistvii spin-mechenogo analoga metatsina s butirilkholinésterazoi.

Interaction between spin-labeled methacyne (I) and butyrylcholinesterase (BChE) was studied by ESR and enzyme kinetic methods. The compound (I) was shown to be a competitive reversible inhibitor, the value of Ki appeared to be 1.3 X 10(-5) M. Insertion of nitroxyl fragment in the methacyne molecule results in a two-fold increase of its inhibitory activity. The ESR spectrum of the enzyme-inhibitor complex was registered. This complex dissociates under the action of eserine, tetramethylammonium and hexamethonium. Scatchard plot reveals two different types of binding sites with Kdiss values 1.5 X 10(-5) M and 2.6 X 10(-4) M. One type of binding sites is identified as the enzyme active centre. The restricted motion of (I) in complex with BChE proves the assumption that the enzyme active centre is located in the split of macromolecule surface.

[Binding of spin-labeled palmitic acid by bovine serum albumin]. / Sviazyvanie spin-mechenoi pal'mitinovoi kisloty bych'im syvorotochnym al'buminom.

Investigation of the binding of spin-labeled palmitic acid and its esters with bovine serum albumin is presented. Both probes are shown to bind strictly to strongly and loosely binding centers of protein. The quantitative study allows to determine the binding constant and number of protein binding sites. It is found that the total binding constant for esterified derivative is several times as much as that of palmitic acid.

[The effect of orthofen (diclofenac) on the embryogenesis of laboratory mice]. / Vliianie ortofena (diklofenaka) na émbriogenez laboratornykh myshei.

The effect of ortophen on the embryogenesis of laboratory mice of two inbred lines BALB/c and C57B1/6 and of outbred stock NMR was studied. The drug was injected intraperitoneally on days 1-6, 6-16, and 16-18 of pregnancy in a therapeutic dose (1 mg/g) and 10-fold doses. General development of the fetuses was delayed and the condition of the pregnant females of both lines and the stock deteriorated on injection of the drug in a 10-fold dose on days 16-18 of pregnancy. BALB/c mice proved to be most sensitive to the unfavorable effects of sodium diclophenac.

[Target delivery of functional genes in gene therapy using carbohydrate containing vectors]. / Adresnaia dostavka funktsional'nykh genov v genoterapii s pomoshch'iu uglevod-soderzhashchikh vektorov.

Various aspects of use of specific interactions with a participation of carbohydrate and oligosaccharide ligands to increase an efficiency of gene transfer into eukaryotic cells (including in vivo experiments) are considered in details. Data on addressed gene delivery with applying carbohydrate-containing ligands (such as asialoglycoproteins and galactosides) are discussed in the paper. Results on the usage of glycoside ligands, containing lactose, mannose, glucose residues, for receptor-mediated gene transfer, are analysed. Special attention is paid to application of chitosans for functional gene transfer into eukaryotic cells, which is considered by authors as a case of receptor-mediated gene transfer. It is notice that neo-oligosaccharide vectors, recognizing surface lectins, represent very perspective type of gene delivery systems.

[Nonviral methods of gene transfer in gene therapy]. / Nevirusnye metody perenosa genov v gennoi terapii.

A number of viral and non-viral vector systems have been developed nowadays for gene therapy applications. The advantages and shortcomings of the following non-viral methods of transfection are discussed in this review: calcium phosphate technique, ballistic transfection using "gene gun", electroporation, microinjection into the nucleus, receptor-mediated gene transfer, and artificial macromolecular complexes (polycations, hydrophobic polycations, polymers, cationic and neutral liposomes). Special attention is paid to methods of lipofection based on the usage of cationic and neutral liposomes as well as targeted gene delivery with the emphasis on the works which were out in author's laboratories.

[Nonviral gene transfer in vivo in gene therapy]. / Nevirusnyi perenos genov in vivo v gennoi terapii.

Recent findings connected with in vivo use of artificial macromolecular complexes (genosomes) for functional gene transfer and delivery are discussed in the paper. Non-viral methods are the most safe for the purpose of human gene delivery. The cationic liposomes containing cholesterol are the most suitable for this purpose, because they possess high biodegradability and stability in blood stream. The DNA-liposome complexes should: (i) contain DNA in the condition at most protected from environmental influence, (ii) be rather homogeneous and of small size (40-80 nm). Injections of complexes into blood are the most effective; in a respect of organospecifity may be achieved by appropriate ligand selection. It is the most perspectively to increase the expression level by combining liposomes with viral peptides.