Fundamentals of protein chemistry at the Institute of Biomedical Chemistry.

Eighty years ago, the Institute of Biomedical Chemistry (IBMC) initially known as the Institute of Biological and Medical Chemistry of the Academy of Sciences of the USSR was founded. During the first decades significant studies were performed; they not only contributed to a deeper understanding of biochemical processes in the living organisms, but also laid the foundation for further development of these fields. The main directions of IBMC were focused on studies of structures of enzymes (primarily various proteases), their substrates and inhibitors, the role of enzymes of carbohydrate metabolism in the development of pathologies, study of the mechanisms of hydrolytic and oxidative-hydrolytic transformation of organic compounds, studies of connective tissue proteins, including collagens, study of amino acid metabolism.

Influence of amino acid and N-terminal protection residue structures on peptide p-nitroanilide adsorption on polystyrene-based support.

Polystyrene-based support Bio-Beads SM-2 was employed for desalting peptide-p-nitroanilides from Oxone. Neither tosyl, 9-fluorenyl(methoxycarbonyl), p-nitroanilide groups nor indolyl or p-hydroxyphenyl side-chains of Trp and Tyr ensured an efficient adsorption of peptide-p-nitroanilides onto Bio-Beads SM-2. Only unsubstituted phenyl-containing protection groups (carbobenzoxy or benzoyl) and Phe residues provided the adsorption of peptides on Bio-Beads SM-2 and their efficient desalting.

Preparation of Affinity Purified Antibodies against ε-Glutaryl-Lysine Residues in Proteins for Investigation of Glutarylated Proteins in Animal Tissues.

The glutarylation of lysine residues in proteins attracts attention as a possible mechanism of metabolic regulation, perturbed in pathologies. The visualization of protein glutarylation by antibodies specific to ε-glutaryl-lysine residues may be particularly useful to reveal pathogenic mutations in the relevant enzymes. We purified such antibodies from the rabbit antiserum, obtained after sequential immunization with two artificially glutarylated proteins, using affinity chromatography on ε-glutaryl-lysine-containing sorbents.

[Application of heat shock proteins as stress markers in aquatic organisms using endemic Baikal amphipods as an example].

When heat shock proteins (HSPs) are used as biomarkers in monitoring studies of aquatic ecosystems, it is necessary to take into account the specificity of synthesis of these proteins in various organisms. This especially applies to endemic species and species with narrow ranges of adaptation for specific conditions in certain water bodies. In this study, we assessed the possibility to use HSPs as molecular stress markers in species with a narrow niche breadth using endemic Baikal amphipods (Crustacea, Amphipoda) as an example.

Evaluation of biochemical responses in Palearctic and Lake Baikal endemic amphipod species exposed to CdCl2.

This study evaluated small heat shock proteins (sHSP) (related to alpha-crystallin) and antioxidant enzymes (POD, peroxidase and CAT, catalase) as possible biomarkers for use in toxicological studies. Biochemical responses to cadmium chloride in two Lake Baikal endemic amphipods (Eulimnogammarus verrucosus, Eulimnogammarus cyaneus) and Palearctic species (Gammarus lacustris) were compared. Our findings showed that cadmium chloride toxicity directly influenced POD activity and sHSP synthesis in all amphipod species.

Natural organic matter (NOM) induces oxidative stress in freshwater amphipods Gammarus lacustris Sars and Gammarus tigrinus (Sexton).

Humic substances comprise the majority of natural organic matter (NOM) on Earth, including dissolved organic matter in freshwater systems. Recent studies show that these substances directly interact with aquatic organisms as chemical stressors. The aim of the present study was to investigate the mode of action of dissolved NOM on the freshwater amphipods Gammarus lacustris Sars and Gammarus tigrinus (Sexton), and in particular, to determine if NOM induces or promotes internal oxidative stress.

Specific antioxidant reactions to oxidative stress promoted by natural organic matter in two amphipod species from Lake Baikal.

Aquatic organisms are exposed to a variety of natural chemical stressors such as humic substances. The aim of this study was to investigate the mode of action of natural organic matter (NOM, roughly 80% of which is humic substances) on two freshwater amphipods from Lake Baikal, Eulimnogammarus verrucosus (Gerstf.) and Eulimnogammarus cyaneus (Dyb.

Non-phosphorylating bypass of the plant mitochondrial respiratory chain by stress protein CSP 310.

Recently, it has been reported that the cold-stress protein CSP 310, discovered in the cytoplasm of cold-resistant winter cereals, causes uncoupling of oxidative phosphorylation during cold stress. To understand how the uncoupling mechanism of CSP differs from that of cyanide-insensitive alternative oxidase and plant mitochondrial uncoupling protein, we determined the effect of respiratory-chain inhibition on winter wheat (Triticum aestivum L. cv.

[An analysis of using the graphic monitoring of ventilation for an optimal choice of respiratory management parameters in patients with acute respiratory distress syndrome].

The potentialities of graphic ventilation monitoring (graphic monitor "Servo Screen 390", Siemens Elema, Sweden) were analyzed for optimizing the respiratory management parameters in 48 obstetric and gynecology patients with acute respiratory distress syndrome (ARDS). The ventilation loops and curves, ALV parameters, mechanical lung properties, gas blood composition and gas indices were dynamically evaluated during examination stages. The graphic ventilation monitoring, when used for respiratory management in patients with ARDS, provides for optimizing, in the real time mode, the PEEP and Vt levels, which is in line with the AVL "safety" concept.

[Diagnostics and correction of thrombohemorrhagic impairments in patients with purulent-septic complications in obstetrics and gynecology].

The clinical course of thrombohemorrhagic complications was studied in 56 patients, aged 18 to 38, with sepsis, which developed due to obstetric and gynecologic processes on days 1, 3, 5, 7 and 10 after surgery. The patients were treated, 1999-2003, at the obstetric intensive-care and consultation center, MUZA Maternity Hospital No. 1, Krasnoyarsk.

Influence of stress protein CSP 310 and antiserum against this protein on oxygen uptake, lipid peroxidation, and temperature of winter wheat seedling shoots during cold stress.

It is determined that infiltration of winter wheat seedling shoots by anti-CSP 310 antiserum caused a significant decrease of oxygen uptake in winter wheat shoots during short-term cold stress. On the other hand, infiltration of winter wheat seedling shoots by stress protein CSP 310 caused an increase of oxygen consumption. The comparison of the influence of infiltration of winter wheat shoots by CSP 310 and anti-CSP 310 antiserum on the rate of lipid peroxidation showed that, if infiltration by CSP 310 caused a decrease of conjugated diene formation, infiltration by anti-CSP 310 antiserum did not cause any significant changes in the rate of lipid peroxidation.

Stress-induced protein CSP 310: a third uncoupling system in plants.

Addition of the cold-stress-related protein CSP 310 to mitochondria isolated from winter wheat ( Triticum aestivum L. cv. Zalarinka), winter rye ( Secale cereale L.

Influence of CSP 310 and CSP 310-like proteins from cereals on mitochondrial energetic activity and lipid peroxidation in vitro and in vivo.

Background: The development of chilling and freezing injury symptoms in plants is known to frequently coincide with peroxidation of free fatty acids. Mitochondria are one of the major sources of reactive oxygen species during cold stress. Recently it has been suggested that uncoupling of oxidation and phosphorylation in mitochondria during oxidative stress can decrease ROS formation by mitochondrial respiratory chain generation.

Stress protein CSP 310 causes oxidation and phosphorylation uncoupling during low-temperature stress only in cereal but not in dycotyledon mitochondria.

It was found that an addition of antiserum obtained against stress protein 310 kD increased coupling of oxidation and phosphorylation in mitochondria isolated from cold-stress winter rye shoots and had no influence on dycotiledon mitochondria (pumpkins and sunflower). The data obtained showed a difference between molecular weights of dycotyledon polypeptides with immunochemical affinity to CSP 310 and CSP 310 subunits. It was shown that low-temperaturestress caused a transition to a low-energy state ("cold uncoupling") of free from endogenous free fatty acid cereal mitochondria.

An influence of antiserum against winter wheat stress uncoupling protein, CSP 310, on energetic activity of some plant species mitochondria.

It is determined that addition of an anti-CSP 310 antiserum to isolated mitochondria of cereals (winter rye, winter wheat, and maize) caused an increasing of mitochondrial respiratory control. In a similar manner, addition of this antiserum to isolated pea mitochondria did not cause this effect. It is shown that coupling effect of antiserum is not dependent upon the presence of bovine serum albumin in mitochondria incubation media.

An influence of stress protein CSP 310 and antiserum against this protein on lipid peroxidation in cereal mitochondria.

It is determined that an addition of an anti-CSP 310 antiserum to isolated winter wheat and maize mitochondria caused more significant increasing of spontaneous lipid peroxidation than the addition of stress protein CSP 310. It is shown that, at function of different mitochondrial respiratory chain complexes, the lipid peroxidation in winter wheat and maize mitochondria take place with different intensities. Under the functioning of mitochondrial respiratory chain complex IV, the maximum output of lipid peroxidation products, dienic conjugates is detected.

The search for proteins with immunochemical affinity to plant stress proteins at cold-adapted endemic Baikal fishes.

The search for proteins with immunochemical affinity to plant stress proteins in endemic Baikal fishes shows the presence of proteins, immunochemically related to plant heat-stabile proteins and plant uncoupling protein CSP 310. Western blotting showed that among the native cytoplasmic proteins of endemic Baikal fishes there are proteins immunochemically related to heat-stabile plant proteins with molecular weights about 480, 200-290, 150, 140 and about 90-100kD. SDS-electrophoresis showed the presence of polypeptides with molecular weights 23, 17 and 14kD in all species investigated and an additional 35kD polypeptide in Cottocomephorus grewingki.

The comparison of uncoupling activity of constituently synthesised and stress-induced forms of winter rye stress uncoupling protein CSP 310.

A difference between the uncoupling action of constituently synthesised and stress-induced forms of winter rye stress uncoupling protein CSP 310 on winter wheat mitochondria in vitro was found from the initiation of incubation. The uncoupling activity of CSP 310 depended on its concentration in the incubation media. The addition of anti-CSP 310 antiserum to isolated mitochondria from stressed winter rye shoots caused coupling of oxidation and phosphorylation.

Complex I of winter wheat mitochondria respiratory chain is the most sensitive to uncoupling action of plant stress-related uncoupling protein CSP 310.

The influence of stress uncoupling protein CSP 310 on functional stability of different mitochondrial respiratory chain complexes was analysed using various substrates of the tricarboxylic acid cycle. Complex I was the most sensitive to CSP 310 uncoupling action whilst other complexes were more stabile. It is proposed that the key point of CSP 310 uncoupling action is complex I of plant mitochondrial respiratory chain.

Plant stress-related uncoupling protein CSP 310 caused lipid peroxidation in winter wheat mitochondria under chilling stress.

The effect of CSP 310 on lipid peroxidation in winter wheat mitochondria was studied by the measurement of primary lipid peroxidation products - dienic conjugates. It was found that some concentrations of CSP 310 caused lipid peroxidation in isolated winter wheat mitochondria in all systems investigated at different concentrations during chilling stress.

[Characteristics of hemostatic disorders in septic shock in children].

Changes at different stages of coagulation cascade have been assessed during intensive therapy of septic shock in 40 children aged 1 to 14 years. Progressing septic shock is accompanied by chronometric and structural hypocoagulation with potential hypercoagulation in transfer samples, thrombocytopenia and thrombocytopathy. Terminal stages of septic shock are characterized by profound hypocoagulation without potential hypercoagulation, predominance of antithrombin and antiaggregant blood activity with persistent depletion of antithrombin III and plasminogen.