[The biphasic nature of the phenomenon of structural adaptational stabilization during the long-term adaptation of the body to stress]. / Dvukhfaznyi kharakter fenomena adaptatsionnoi stabilizatsii struktur v protsesse dlitel'noi adaptatsii organizma k stressu.

During long-term (28 days) adaptation of the body to stress, the structural adaptational stabilization phenomenon (SASP) has a marked two-phase pattern: (1) SASP formation during nearly 14 days when protein transcription in heat shock was activated, hsp 70 was substantially accumulated and cardiac resistance to reperfusion damage drastically increased and (2) SASP reduction occurring despite continuous adaptation. By day 28 myocardial adaptation showed only two hsp 70 isoforms of the five revealed on day 14 of stress adaptation. Correspondingly, by day 28, the anticontractural effect of adaptation and its capacity of retaining the high contraction amplitude turned out to vanish in reperfusion. Moreover, ventricular fibrillation and ventricular tachycardia were observed in the control animal hearts during reperfusion, whereas these arrhythmias were seen only in 25 and 50% cases on days 14 and 28, respectively.

[The generalized accumulation of stress proteins during body adaptation to stress exposures]. / Generalizovannoe nakoplenie stress-belkov pri adaptatsii organizma k stressornym vozdeistviiam.

It has been found that accumulation of stress proteins during adaptation to immobilization stress occurs not only in the heart, but in the brain (2 isoforms in the cytoplasm and 2 in the nucleus) and in the liver (1 isoform in the cytoplasm and 1 in the nucleus of hepatocytes). Thus, in heat shock the activation of protein synthesis is generalized during the body's adaptation to stress. This may underlie the generalization of the structural adaptational stabilization phenomenon and the formation of an adaptative defense of the whole organism.

[Isoform pattern of inducible HSP 70 in the rat myocardium after heat shock]. / Izoformnyi sostav indutsibel'nykh HSP 70 v miokarde krysy posle teplovogo shoka.

The hsp 70 content was measured in the myocardium of a control rat group, in the group of rats 24 hours following a heat shock and in the group of rats 48 hours after a heat shock. In 24 hours after the heat shock, a major inducible hsp 70 with molecular weight of 71 kDa and pI about 5.8 occurred which was utterly absent in myocardial cytosol from control animals. In addition, there was an increase in polypeptide fraction with molecular weight of 73 kDa and pI about 5.6 (HSX73). In 48 hours after the heat shock, first the inducible hsp 70 with molecular weight of 71 kDa and pI about 5.8 disappeared which was found in 24 hours; secondly, HSX73 decreased to the control level and, thirdly, several isoforms pronouncedly accumulated with molecular weight of about 71 kDa and pI ranging from 5.9 to 6.3. Thus, the isoform composition of stress proteins induced by heat shock strongly depends on the time after the stress exposure. Furthermore, the accumulation of more acidic isoforms precedes the accumulation of alkaline ones.

[Cardioprotective effects of adaptation to restraint stress and hypoxia]. / Kardioprotektornye éffekty adaptatsii k immobilizatsionnym stressam i gipoksii.

Isolated rat heart experiments have revealed that restraint stress adaptation results in enhanced resistance of the isolated heart to reperfusion. There is also a higher resistance to the autolysis of the organelles isolated from the hearts of stress-adapted animals. This complex of changes is designated as a phenomenon of adaptive stabilization of structures (PhASS). The phenomenon developing in restraint stress adaptation substantially limits arrhythmias, contracture, contraction amplitude depression, and creatine kinase release into the perfusate in thermal damage to the isolated rat heart. Simultaneously, PhASS is accompanied by a multiple increase in five hsp70 isoforms with pI 5.8-6.3 in cytosol and two isoforms with pI about 6.3 in the nucleoplasm. Only two hsp70 isoforms with pI about 5.8 accumulate solely in cytosol during adaptation to intermittent hypoxia. Consistently, the resistance of Ca(2+)-pump and nuclear DNA remains unchanged and the protection against reperfusion and thermal damage are several times less pronounced.

[Phenomenon of adapted stabilization of structures and the role of heat shock proteins in its mechanism]. / Phenomen adaptatsionnoi stabilizatsii struktur i rol' belkov teplovogoshoka v ego mekhanizme.

Mechanisms, responsible for distinct increase in rat heart tissue resistance to reperfusion paradox and toxic concentrations of catecholamines and Ca2+, were developed in the heart after repeated stress actions. Sarcoplasmic reticulum and mitochondria isolated from heart cells of adapted animals were highly resistant to autolysis, while nuclei--to the impairing effect of single-strand exogenous DNA. These alterations were designed as a phenomenon of adaptive stabilization of structures (PhASS). Accumulation of heat shock proteins was found to be of importance in mechanisms of PhASS. Development of PhASS was accompanied by an increase in resistance of myocardium to ischemic necrosis.

[Adaptation of the body to stress effects increases resistance of nuclear DNA of cardiac cells due to accumulation of heat shock proteins in the nucleus]. / Adaptatsiia organizma k stressornym vozdeistviiam povyshaet ustoichivost' iadernoi DNK serdechnykh kletok za schet nakopleniia belkov teplovogo shoka v iadre.

It was shown that adaptation to stress exposure increased the resistance of nuclear DNA in myocardial cells to the damaging action of exogenous one-chain DNA (50 micrograms/ml). This protective effect was accompanied by a pronounced accumulation of heat shock proteins (hsp) 70 in nucleoplasm of myocardial cells from adapted animals. Possible mechanism of the DNA protective effect of adaptation and the role of hsp 70 are under discussion.

[Systematic approach to studying proteins from human platelets. Two-dimensional mapping]. / Sistamaticheskii podkhod k izucheniiu belkov trombotsitov cheloveka. Postroenie dvumernoi karty.

Total proteins of human platelets and their membranes were studied by two-dimensional electrophoresis using the lysis in sodium dodecyl sulfate solution. Analysis of platelets from 30 donors revealed the presence of 105 repeating fractions on the electrophoregrams. A two-dimensional map of platelet proteins in the molecular mass versus relative electrophoretic mobility plot was constructed. This map made it possible to localize membrane proteins and albumin as well as a protein immunologically related to phenylalanine hydroxylase. Electrophoretic variants of 13 platelet polypeptides were identified.

[Dynamics of DNA-binding activity of cytoplasmic proteins in autoproteolysis]. / Dinamika DNK-sviazyvaiushchei aktivnosti tsitoplazmaticheskikh belkov pri autoproteolize.

The quality proteolysis changing of DNA-binding proteins of cytosol mice liver was studied by affinity chromatography on DNA-cellulose. It is shown that neutral proteolysis leads to the second peak of DNA binding.

[The use of highly sensitive immunoblotting for detecting the phenylalanine hydroxylase antigen in human platelets]. / Primenenie immunoblottinga povyshennoi chuvstvitel'nosti dlia vyiavleniia fenilalaningidroksilaznogo antigena v trombotsitakh cheloveka.

Incubation of nitrocellulose filters containing proteins in solutions of organic alcohols (ethanol, methanol, isopropanol) enabled to increase the immunoblotting sensitivity after denaturating electrophoresis in presence of SDS. Maximal elevation of the label sensitivity (4-6-fold) was observed after incubation of these filters in 30% isopropanol within 2 hrs. The effect of sensitivity elevation appears to be caused by the antigen renaturation due to SDS washing off. The modified procedure of immunoblotting allowed to detect phenylalanine hydroxylase antigen in human thrombocytes. The antigen had electrophoretic mobility similar to that of phenylalanine hydroxylase from liver tissue; its concentration constituted less than 0.1 microgram per 1 mg of protein in thrombocytes.

[Detection of phenylalanine hydroxylase protein in human platelets. Immunochemical detection]. / Obnaruzhenie belka fenilalaningidroksilazy v trombotsitakh cheloveka. Immunokhimicheskaia identifikatsiia.

A protein reactive with anti-phenylalanine hydroxylase monoclonal antibody PH8 has been recovered from human platelet extracts. Two bands corresponding to molecular masses of about 60 kDa and 55 kDa were revealed by immunoblotting after electrophoresis according to Laemmli. Using the same antibody, a single band with a molecular mass of 60 kDa was demonstrated in extracts from human pineal gland; two similar antigens were found in human liver extracts and no antigen was found in adrenal gland extracts. Monoclonal antibodies, PH1 and PH3, did not react with these antigens during immunoblotting. Monoclonal antibodies, PH7 and PH9, reacted with the 55 kDa antigen in platelet extracts. The antigen content in platelet extracts was measured by ELISA with monoclonal antibodies relative to its content in the liver. The antigen content in platelet extracts was about 100 times as low as that in liver extracts and amounted to 100 ng/mg of protein. These findings suggest that the phenylalanine hydroxylase antigen is present in human platelets.

[Effect of gamma-oxybutyrate on energy metabolism and the cyclic nucleotide level in nerve tissue in experimental intracerebral hemorrhage]. / Vliianie gamma-oksibutirata na énergeticheskii metabolizm i uroven' tsiklicheskikh nukleotidov v nervnoi tkani pri éksperimental'nom vnutrimozgovom krovoizliianii.

Experimental intracerebral hemorrhage (EICH) was shown to cause a disorder of the functional state of the brain mitochondria. The rate of oxidation of NAD- and FAD-linked substrates in 3 and 4 states decreased 1 and 24 hours after hemorrhage. An injection of gamma-hydroxybutyrate 1 hour before decapitation increased the rate of mitochondrial respiration. EICH was found to increase cGMP level and to decrease cAMP level. A combined action of hemorrhage and gamma-hydroxybutyrate produced a greater lowering of cAMP level and an elevation of cGMP level.