[PHOSPHOLIPIDS, FATTY ACIDS AND HEMOGLOBIN OF ERYTHROCYTES IN RAT BLOOD UNDER STRESS CONDITIONS (SWIMMING AT A LOW TEMPERATURE)].

The phospholipid and fatty acid content of erythrocytes in rat blood was studied after exposure to stress - swimming up to drowning. This stress was found to increase the content of phospholipids cha- racteristic for the membrane outer layer (phosphatidilcholine by 13 % and sphingomyelin by 23 %). At the same time a decrease in the content of acid phospholipids belonging to membrane inner layer was ob- served (phosphatidylethanolamine by 16 %, phosphatidyl serine by 14 % and monophosphoinositide by 23 %). The data obtained indicate that erythrocyte membrane under stress conditions undergoes certain structural changes which seem to have an effect on its functional properties. At the same time it was fo- und that the content of saturated, unsaturated and ounsaturation index? of fatty acids does not undergo es- sential changes under stress exposure studied thereby, possibly, compensating maintenance of functional properties of the erythrocyte membrane under conditions of the impaired phospholipid structure. On the basis of examining absorption spectra of lipid extracts it was found that stress evokes a two-fold spectrum growth in the range of 390-410 nm appropriate to the hem. The fact of appearance of the hem in the ext- ract indicates to saponification of hemoglobin evoked by changed pH of the erythrocyte internal environ- ment. Actually, in the process of lipid extraction the hemoglobim passes into a disordered state due to the effects of not only temperature and environmental pH but also due to those of organic solvents possessing lower, than water, ability to form hydrogen bonds. Probably, transesterification of a minor portion of phospholipids occurs during their extraction from erythrocytes with the chloroform-methanol mixture.

[PHOSPHOLIPIDS AND FATTY ACIDS IN ERYTHROCYTES OF THE LAMPREY LAMPETRA FLUVIATILIS DURING AUTUMN PRESPAWNING PERIOD AND THE ABSORPTION SPECTRUM OF THEIR LIPID EXTRACT].

The content of some classes of phospholipids and their fatty acid composition in erythrocytes of the lamprey Lampetrafluviatilis during the autumn period of its prespawning migration are investigated. It is found that the phospholipid spectrum of erythrocytes of the lamprey, the oldest representative of vertebrates, is similar to that of many mammals. A four-fold prevalence of phosphatidilcholine content over sphingomyelin content as well as prevalence of (ω3-acids over ω6-acids indicates the <> of lamprey's erythrocyte membranes - an important indicator of deformational ability of lamprey's erythrocytes. Phosphatidilethanol amine and its plasmalogenic form are the most unsaturated phospholipids (their unsaturation indices are 230 and 342, correspondingly). Phosphatidilcholine is the most saturated one (UI is 167). It is found that the basic acid indicators characterizing the fluidity of erythrocyte membranes remain unchanged during the whole period of prespawning migration of lampreys up to spawning. The blood contains several buffer systems, in particular, membrane phospholipids which neutralize acids and alkali incoming into the blood. In the process of organism life a change of pH inside erythrocytes occurs. One can suppose that the base of the system associated with buffer properties of the blood is water dissociation. Inside thin vessels of the circulatory system the hemoglobin attaches and returns molecules of oxygen due to interaction of the buffer systems with water. The property of water to dissociate as well as ion transfer produce in erythrocytes, lying within narrow vessels of the circulatory system, a local pH alteration allowing displacing/attaching the molecule of oxygen from hemoglobin.

[Fatty acid composition of phospholipids of erythrocytes of lamprey, frog, rat, and absorption spectra of their lipid extracts].

The work deals with study of content and fatty acid composition of phospholipids as well as of absorption spectra of lipid extracts of blood erythrocytes poikilothermal and homoiothermal animals of different evolutionary levels. Objects of study were poikilothermal lamprey (Lampetra fluviatilis) consuming oxygen from water and the common frog (Rana temporatia) consuming it both from water and from air. Homoiothermal animals were white rats (Rattus rattus) inhabiting in the air medium. The animals were studied at the winter-spring periods. There was established the twofold predominance of the phospholipid content in the lamprey plasma as compared with erythrocytes. In frog and rat the reverse ratio was observed. Based on study of the fatty acid composition of erythrocyte phospholipids it is suggested the higher density of membranes of lamprey as compared with frog membranes. As to fatty acides of the rat blood erythrocytic fraction, they turned out to be less diverse, with almost twofold predominance of saturated over unsaturated acids and not containing the long-chained (C22) Ω3 acids. All this leads to the low unsaturation index and, accordingly, to a dense packing of fatty acids in membrane structures of rat erythrocytes. Mechanism of reversible binding of O2 molecules by hemoglobin in erythrocytes is discussed. The mechanism of interaction of O2 molecules with water molecules is likely to interfere with exchange interaction electrons of hemoglobin iron atoms and oxygen molecule. This confirms our obtained absorption spectra showing that in the lipid extract practically not containing water the heme isolated from erythrocytes is converted to hemin.

[Effect of stress actions on some hematologic and biochemical parameters of rat blood and on energetic intermolecular interactions in lipid extract under effect of light radiation].

Comparative study has been carried of effect of the three-day long starvation, running, and their combination on morphological parameters of rat blood, lipid metabolism, and activity of blood Na,K-ATPase. Different effect has been shown of these stress factors on the blood erythrocyte composition. Starvation is accompanied by the most pronounced release of stored erythrocyte into blood, which results in a significant decrease both of the total amount of reticulocytes and the complete absence of reticulocytes of the I stage of maturity (the youngest). The running on treadmill led to a significant increase of the total amount of blood reticulocytes and to multiple increase of immature reticulocytes (RC-I and RC-II), which can indicate some stress of the bone marrow erythroid stem line. The curve of acid resistance of blood reticulocytes has shown the animal to experience the greatest stress at a combination of starvation and running. Starvation and running produced different effects on blood lipid characteristics. The content of triacylglycerides (TAG) in blood rose by 40% at starvation and decreased by 30% at running, a similar tendency being found for index of atherogeneity. The fatty acid composition of blood phospholipids at running and its combination with starvation practically did not differ from control. A change of Na,K-ATPase, which is so characteristic of reaction to various kinds of stress, sharply fell at starvation (by 22%), but increased at running (by 13%) and decreased markedly at combination of these actions. Absorption spectra of lipid extracts of the whole blood of the rats submitted to various stress actions showed that extracted from blood (at different amount depending on the kind of action) is an organic substance with coupled bonds, which absorbs light in the diapason of 360-620 nm. The absorption of light in the diapason of 400-410 nm has been found to belong to the Soret band of ferroheme and ferriheme. The shift of the Soret band indicates electron transitions in the iron cation. By the change and disappearance of the Soret band, it is possible to judge about the processes occurring in the lipid extract. The disappearance of the Soret band in the lipid extract indicates formation in it of steady radicals as a result of the ferriheme disintegration due to accumulation of energy in porphyrin, which does not seem to occur in the blood cell membranes. The iron atom in the ferriheme molecule is known to accept electron and yields a part of energy probably to porphyrin. Then ferriheme yields electron and becomes ferriheme with excess of energy in porphyrin. Hence, at admission of the next electron to the iron atom the porphyrin molecule is to get rid of the energy obtained earlier to prevent its disintegration. The heme is possible to be an accumulator and distributor of energy in tissue.

[Bounds of change in unsaturation index of fatty acid composition of phospholipids at adaptation of molluscs to biogenic and abiogenic factors of external medium].

Comparative study of fatty acid composition of total phospholipids, as well as of phosphatidylcholine and phosphatidylethanolamine from hepatopancreas and leg muscle was performed on several representatives of gasteropods (Gastropoda) molluscs and bivalve (Bivalvia) mussel (Mytilus edulus). The objects of our study were marine litorins (Littorina saxsatilis) adapted to different temperature conditions of White Sea and Barents Sea, freshwater lymnaea (Lymnaea stagnalis) infested by Trematoda and mussels from White Sea and Black Sea. It was shown that depending on the existence conditions of studied tissue or lipid, the maximal change is observed in the percentage of saturated acids (4-83 %), the percentage of unsaturated acids was less expressed (1-14 %) and the changes in unsaturation index (UI) did not exceed 20 % on average. It was supposed that observed quantitative bounds of UI change under the action of different external factors is utmost for maintenance of membrane fluidity necessary for normal vital activity of cell, particularly in studied ectothermic molluscs.

[Effect of total hypothermia on the fatty acid composition of blood phospholipids of rats and sousliks and light irradiation on chemical processes in lipid extract].

Effect of hypothermia on the fatty acid composition of rat and souslik blood phospholipids is studied. Different reaction of these animals to cooling is revealed: in rats no changes were observed in the fatty acid composition of blood phospholipids, whereas in the hibernating there were significant changes in the content of individual fatty acids (FA). The content of monoenic acids in sousliks decreased almost by 50%, while the content of saturated acid (C18) and of polyenic acids C18 : 2omega6 and C20 : 4omega6 rose significantly. Such changes seem to be the mechanism that promotes maintenance of the organism viability under conditions of a decreased level of metabolism, heart rhythm, and body temperature and is evolutionarily acquired. At the same time, the observed changes in the content of individual FA do not lead to sharp changes in such integrative parameters as the total non-saturation of phospholipids, which determines liquid properties of chylomicrons and other lipolipoprotein transport particles of the souslik blood. There are studied absorption spectra of blood lipid extracts of rats and sousliks under effect of light as well as effect of light upon the FA composition of lipid extracts of these animals. The FA composition of lipid extracts has been established to remain practically constant, whereas the character of changes of spectra under action of light indicates the presence in the extracts of oxidation-reduction reactions. The obtained data allow suggesting that in the lipid extract there occurs cooperation both of the phospholipid molecules themselves and of them with other organic molecules, which makes it possible for fatty acids to participate in processes of transport both of electrons and of protons. This novel role of FA as a participant of the electron transfer might probably be extrapolated to chemical reactions (processes) occurring inside the membrane.

[Fatty acids of phospholipids of brain cell nuclei in rat ontogenesis].

Experiments were carried out on the 19-day old rat embryos, the 5- and 45-day old rat pups, and the 1.4-1.5-year old rats. Phospholipids and their fatty acid composition in brain cell nuclei were studied using methods of extraction, two-dimensional thin-layer chromatography, and spectrofluorimetry. In the course of ontogenesis, the percentage of different classes of nuclear phospholipids was changed; at the postembryonic period, the unsaturation index of lipids (phosphatidylcholine and phosphatidylethanolamine) and the content of unsaturated (especially polyenic) fatty acids decreased. Microviscosity of nuclear membranes increased; this changes were also shown earlier in phylogenesis of vertebrates. Thus, the facts revealed in the present work correspond to the recapitulation law. It is suggested that such change of lipid ratio and of composition of their fatty acids as well as of the membrane microviscosity serve for regulation of functions of membrane proteins and have adaptive character.

[Significance of electron interactions of fatty acids of phospholipid molecules in the organism adaptation to habitation temperature].

Data in the fatty acid composition of muscle tissue phospholipids of some representatives of gastropod molluscs (Gastropoda) have been presented for the first time. In the lake phytophagues Lymnaea stagnalis and Lymnaea ovata the long-chained C22-acid was not detected, whereas in the predator common whelk Buccinum undatum, C22:6omega3 was present. Comparison of absorption spectra (240-720 nm) of lipid extracts of the studied invertebrates and of rat has been performed. The obtained data are discussed from the point of view of participation of pi-electrons of phospholipid fatty acid molecules in adaptation of membranes to the habitation temperature, which arises owing to interelectron attraction and to the process of formation of Cooper's pairs.

[About effect of habitat and motor activity of molluscs on fatty acid composition of triglycerides and phospholipids].

A comparative analysis of fatty acids (FA) in neutral and phospholipids of digestive gland and pedal muscle has been performed in molluscs from various ecological groups differing by belonging to sea or fresh water, trophic types or the associated motor activity. In freshwater pulmonary gastropods Lymnaea stagnalis and Limnaea ovalis and marine prosobranchial molluscs Buccinum undatum and Littorina littorea the total content of omega3-acids in phospholipids of the studied tissues differed more than twice, predominantly due to the combined effect of temperature and salinity of the habitat. The lower viscosity of cell membranes in marine species (omega3/omega6 < 1) is determined to the greatest degree by the presence of eicosapentaenoic acid that accounts for 22-25 % of the FA sum in marine species. Comparison of the molluscs by their trophic belonging has revealed the presence of linoleic acid in triglycerides in digestive glands of phytophages (8-12 %), but the practically complete absence of this acid in the predator B. undulatum (<0.8 %. By mobility, L. littorea inhabiting the high-low tide littoral was inferior to freshwater pulmonary gastropods and to marine predator, as it stops moving twice a day during the low tide. In phospholipids of pedal muscle of this mollusc the amount of long-chain polyunsaturated C:22 FA was 3-6 times lower than that in other studied species, which might possibly indicate the role of these acids in functioning of the pedal muscle contractile tissue. On the whole, use of the FA characteristics as parameters determining belonging to certain ecological group requires a certain caution due to a complex action of biotic and abiotic factors on the animal metabolism. The exception is the omega3/omega6 ratio in total phospholipids of freshwater and marine gastropods.

[About paired correlation of pi-electrons tissue lipid extracts of animals of different evolutionary levels].

With aid of optical methods, the presence of the paired correlations of pi-electrons has been revealed in phospholipids as well as in triacylglyceride molecules. Used for analysis were lipid extracts of individual representatives of animals of various evolutionary levels--cartilaginous and bony fish and mammals differing by the content of unsaturated fatty acids in lipids. It has been established that the necessary condition for formation of electron pairs is interaction of lipid molecules with each other. An opinion is put forward that in the liquid crystal structure of the membrane monolayer there are two zones able to form electron pairs--the zone of location of ester bonds and the zone in the region of double bonds. Besides, the paired correlation in the phospholipid molecule electron system is accompanied by the absence of electric resistance of the membrane monolayer, which provides the monolayer superconductivity at low rates of movement of the "electron fluid". It is to be noted that the very fact of the presence of the electron pair implies transfer of energy by small portions, which does not allow excitation of individual phospholipid molecules in the monolayer and promotes stability of the native membrane. Our data agree with the known statements of A. Pulman and B. Pulman that the life dynamicity is determined by dynamicity of the electron cloud in coupled or partially coupled systems.

[Changes produced by trematode larvae in the phospholipid fatty acid composition of digestive gland of the mollusc Littorina saxatilis].

Lipids of the digestive gland of the mollusc Littorina saxatilis from the White and Barents Seas were studied. Changes of its biochemical composition are discussed in the connection with different temperature of the habitat and with infestation with trematode larvae. Comparative analysis of the fatty acid (FA) composition of each of phospholipids in intact molluscs has revealed essential differences. Phosphatidylcholine and monophosphatidylinositol (MPI) FA did not differ in the omega 3/omega 6 ratio, which is due to their tolerance to the temperature factor, whereas more unsaturated phospholipids--phosphatidylethanolamine (FEA), its plasmalogen form (pFEA), and phosphatidylserine--differed 1.5-2 times in the studied molluscs. Predominance of omega 3 acids in the Borents Sea molluscs undoubtedly is due to the lower habitat temperatures, as it provides a higher fluidity of membrane phospholipids. Infestation affected to the greatest degree the quantitative FA composition in pFEA and MPI. At infestation, out of all considered phospholipids, only in MPI there was revealed a threefold decrease of the content of eikosenoic acid C20 : 1, whereas in all other phospholipids, in the contrary, it increased. Monophosphatidylinositols also differed essentially from other phospholipids by the saturated FA amount, which changed the unsaturation index of these phospholipids. Since the functional significance of this minor phospholipid is determined by its participation in the so-called phosphatidylinositol system of the hormonal signal transduction, it seems interesting to elucidate whether an increase of this membrane phospholipid saturation at invasion affects the reflex connection between signals from receptors located in a parasite and enzymatic processes.

[Combined effect of environmental temperature and trematodes on fatty acids composition of lipids of Littorina saxatilis (Olivi 1792) (Gastropoda, Prosobranchia)]. / O sovmestnom vliianii prirodnykh temperatur i trematod na zhirno-kislotnyi sostav lipidov u Littorina saxatilis (Olivi 1792) (Gastropoda, Prosobranchia).

An effect of environmental temperatures and invasion by helminthes larvae on fatty acids composition of digestive gland lipids of marine littoral gastropod Littorina saxatilis from White Sea and Barents Sea was investigated. We have compared gastropods from boreal and subarctic populations. It was found that gastropods from waters of lower temperature have increased omega3/omega6 fatty acids ratio. However, saturation index of individual membrane phospholipids was not affected. Also, content of eicosenoic acid (20:1) in individual phospholipids was not affected by temperature. Invasion increases the omega3/omega6 ratio of common lipids but not the omega3/omega6 ratio of common and individual phospholipids with the exception of phosphatidilcholine of cold water mollusks from Barents Sea that had this ratio doubled. In contrast to temperature, invasion affects the content of eicosenoic acid that was increased in the investigated organs. Adaptability of these effects is discussed regarding parasite-host system.

[Lipids of the bone tissue in bodies of thoracic vertebrae in scoliosis in rabbits]. / Lipidy kostnoi tkani tel pozvonkov grudnogo otdela pozvonochnogo stolba krolikov pri éksperimental'nom skolioze.

The content of total lipids, triglycerides, cholesterol and phospholipids in the bone tissue of the bodies of the thoracic vertebrae was investigated in 4.5-, 7- and 12 month old rabbity. The development of scoliosis in two month old animals was induced by the one-sided blockade of the afferent links of the innervation in thoracic region at the extent of four segments (TH6-9). In: experimental scoliosis there were important alternations in the content of the total lipids comparing with the control which are observed only on the level of the primary and of the anterior compensatory curves in 12 months old animals. The content of triglycerides changed even at early stages of the development of scoliosis. A decrease of triglycerides content was observed at the level of the primary curve in rabbits of all ages with experimental scoliosis. Probably it is due to the alteration of neurotrophic processes in the bone tissue. There were similar age-related changes of triglyceride content in both groups of animals. Phospholipids have absolutely another than triglycerides dynamics of the content changes during the scoliosis development. Time-course of phospholipid changes in scoliosis differed from that of triglycerides. In older rabbits with experimental scoliosis the cholesterol content in the bone tissue of vertebral bodies was reduced by 1.5-2 times while in normal animals it remained almost unchanged. The most substantial changes in cholesterol content of the vertebral bone tissue in rabbits with the experimental scoliosis occur in early age (4.5 months) on the level of the primary curve and the posterior compensatory curve. This may be considered as one of the biochemical indicators of the progressing scoliosis.