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Department for Biochemistry of Cold Adaptation
Research Fellows
of the Department:

Department for Biochemistry of Cold Adaptation was organised in 1989. The department is headed by Prof. Gulevsky A.K., DSc (Biol). One of the aspects of department activity is the study of molecular mechanisms of winter hibernation. The investigation of peculiarities of biomembranes, protein-synthesising apparatus, enzyme functioning will allow to create the theory on a natural adaptation of different organ and tissue cells of hibernating animals under natural hypothermia conditions, and in a practical respect will be an approach for scientific substantiation of a long-term storage of cell suspensions, organs and tissues of homoiothermal animals assigned for transplantation.

Head of the Department
Alexandr K. Gulevskiy
Prof., DSc(Biol)

An important direction of department's investigations is also the study of biomolecules-adaptogenes of cells in poikilothermal and heterothermal animals to cooling. Using the current methods in biochemistry and molecular biology, such as chromatography, gel-filtration, electrophoresis ultracentrifuging, radioisotope analysis we search for biomolecules of cold-resistant animals, modifying physical and chemical properties of intercellular and intracellular medium, antifreezes, proteins of temperature hysteresis, ice proteins-initiators, as well as biomolecules, modifying physical-chemical properties of the most important macromolecular complexes of membranes, enzymes and chromatin towards cell stabilisation to low temperature effect.

As a result of the investigations conducted the erythrocytes of Citellus undulatus ground squirrels, being in hibernation were established to have an increased stability to a stress effect, in particular, to a long-term storage under hypothermia conditions (1 month), that manifested in hemolysis reduction, preservation of membrane barrier properties, ion homeostasis and its controlling systems. The Citellus undulatus erythrocytes were shown to have a maximum resistance in the beginning of baut in January - February.

The application of 1-10 kD protein fraction, isolated from tissues of cold-adapted and hibernating animals, allowed to found out the series of pharmacological effects. Under 10-7 M concentration the fraction causes a manifested antipyretic and antiinflammatory effect, has a protective effect on myocardium of homoiothermal animals (rats) under hypothermic storage conditions, causes a statistically significant reduction of cold sensitivity of skin thermoreceptors, inhibits biosynthesis of proteins, DNA, RNA in cells of Erlich's ascites carcinoma.

The 1-10 kD protein fraction, isolated from the Yakut horse's brain (during winter) within the concentration range from 10-7 M to 3x10-5 M causes the rate decrease in calcium passive transport and an increase in an active transport rate in the inverted sarcolemma vesicles of cardiomyocytes under calcium physiological concentration in the incubation medium. It should result in a decrease in Ca2+ concentration in the intact cardiomyocytes.

Under low temperatures (2-4C) conditions of acclimation of T. molitor cold-resistant bug larvae the biosynthesis of proteins and RNA occurs, that is testified by the inclusion of 14C-aminoacids and 3H-uridine into the proteins and RNA, correspondingly. Specific inhibitors of protein and RNA synthesis: cycloheximide and amanitin were shown to suppress these processes.

During T.molitor larvae acclimation the change in protein composition occurs, in particular, there is a considerable increase in probability of appearance of 65 kD molecular mass protein. The protein-synthesising cell system was found out to take part in the acclimation process.

When studying the lipid synthesis de novo by means of a radioactive marker it was revealed that during low temperature acclimation there were synthesised the fractions of both neutral and polar lipids. There is the most considerable increase in synthesis of neutral lipids, free fatty acids and sterol ethers. Of special note is the fact of lipid synthesis itself under 4-6C, moreover the level of a radioactive precursor inclusion is not quantitatively lower, than under 20-22C, but slightly higher in some cases.

As a result of performed study of cell stabilisation mechanisms to the action of a non-penetrating cryoprotectant and low temperatures there were substantiated the new approaches for a targeted modification of structural and functional cell state at the initial stages before cryopreservation with the aim to increase their resistance to freeze-thawing. These approaches consist in the formation of certain temperature-osmotic conditions when the stabilisation of plasmatic membranes and cytoskeletal proteins occurs. Basing on the principle of "cold stabilisation" there was developed the method without washing-out for erythrocyte cryopreservation under protection of a non-penetrating PEO-1500 cryoprotectant, which allows to increase in a considerable extent their resistance to freeze-thawing and to preserve structural and functional cell indices at a quite high level. This method is more perspective in comparison with the actual ones and can be widely applied in practical medicine.

For the last period in our department there were defended 3 doctor's degree thesis and 8 PhD thesis, 3 collaborators obtained the Professor title. Now the former collaborators of the department for biochemistry of cold adaptation head the chairs in the higher institutions: Prof. Zhegunov G.F. is the Head of the Chair of Chemistry and Biochemistry at Kharkov State Veterinary Academy of the Ministry of Agricultural Industry of Ukraine, Prof. Subbota N.P. is the Head of Valeology Chair at Kharkov Pedagogical University.

Now the department develops a new scientific direction. The investigations deal with a biological activity of cord blood and its component on different biochemical, immunological and other indices of mammalian organism. At the same time it is supposed to increase the adaptive properties of an organism, in particular, to the long-term physical loads.

23, Pereyaslavskaya str., Kharkov, Ukraine 61016
Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine
Tel:Tel: +380 57 373 4143,   373 3807,   373 3039;     Fax: +380 57 373 3084
E-mail: cryo@online.kharkov.ua