[A role of annexin-А5 in pathogenesis of ischemic stroke]. / Rol' anneksina-A5 v patogeneze ishemicheskogo insul'ta.

Objective. To study the association between ischemic stroke (IS) and rs11575945 polymorphism of gene encoding an apoptotic marker annexin-A5 protein (ANXV), to evaluate the serum content of annexin-A5 protein in patients with IS on the first day of stroke compared to healthy subjects and the relationship between the levels of annexin-A5 protein and rs11575945 polymorphism. Material and methods. The study included 94 patients in the acute stage of IS. Polymerase chain reaction with sequence-specific primers and enzyme-linked immunosorbent assay were used. Results and conclusion. ANXV mutant allele frequency was in average 2.6 times higher in patients (p<0.00001) than in healthy subjects, and annexin-A5 mean concentration in the blood serum of IS patients was 3 times higher compared to healthy subjects. Moreover, the concentration of annexin-A5 in the serum of the rs11575945*T mutant allele carriers of the ANXV gene was 4 times higher in comparison to individuals homozygous for the rs11575945C standard allele.

[The involvement of abnormal apoptosis in the disturbance of synaptic plasticity in posttraumatic stress disorder].

Authors determined the levels of marker proteins for apoptosis and synaptic plasticity - annexin-А5 and complexin-2, correspondingly, as well as the proinflammatory cytokine tumor necrosis factor-α (TNF-α) in the blood serum of patients with posttraumatic stress disorder (PTSD) in comparison to healthy controls using the enzyme-linked immunosorbent assay. An analysis of correlations between these parameters was performed. According to the results obtained, the levels of annexin-А5 and complexin-2 were significantly lower, and the levels of TNF-α were significantly higher in PTSD patients. In addition, a positive correlation between the levels of annexin-А5 and complexin-2, on the one hand, and a negative correlation between the levels of annexin-А5 and TNF-α, on the other hand, were detected in PTSD. It has been concluded that PTSD is characterized by the lower apoptosis rate associated with the defects in synaptic plasticity. It is proposed that abnormal apoptosis may be also one of the factors responsible for development of PTSD-associated chronic inflammation.

[Markers of apoptotic dysfunction in schizophrenia].

According to modern concepts, alterations of the apoptosis processes and its genetic regulation are involved in etiopathogenesis of schizophrenia. This is observed on the levels of both the brain and peripheral blood. However, studies in this aspect are on initial stage of development, and molecular and cellular mechanisms of abnormalities of apoptosis in schizophrenia are not clear. In the present study we determined the levels of apoptotic markers, annexin-A5 and ficolin-H proteins, in the serum of patients with chronic and first-episode schizophrenia and healthy controls. The potential association of functional single nucleotide polymorphism rs11575945 (-1C/T) of "Kozak" consensus sequence in the regulatory region of the annexin-A5 gene with schizophrenia was examined. In this study the enzyme linked immunosorbent assay and polymerase chain reaction with allele-specific primers were used. The results suggest that the pathogenesis of schizophrenia is characterized by increase rate of apoptosis, which is more pronounced in case of first-episode neuroleptic-free patients than in case of chronic patients treated with typical neuroleptic haloperidol. It was also shown that rs11575945 polymorphism of the annexin-A5 gene is associated with schizophrenia, and its minor allele is responsible for higher levels of the annexin-A5 protein in the blood and represent one of the risk factors for this disease.

[The influence of aromatic amino acid derivatives on the complement system under ionizing radiation].

Activities of the complement classical, alternative and lectin pathways were determined under conditions of X-radiation in the blood of rats treated and none-treated with synthetic Schiffbase aromatic amino acid derivatives, nicotinyl-L-tyrosinate or nicotinyl-L-tryptophanate, before irradiation. In case of activities of the alternative and lectin pathways no significant changes between irradiated animals and none-irradiated control animals were detected. However, the data obtained demonstrate significantly elevated activity of the classical complement cascade in the blood of irradiated animals (1 day after irradiation), as compared to those none-irradiated. This effect was less pronounced in rats treated with nicotinyl-L-tyrosinate or nicotinyl-L-tryptophanate 1 hour before irradiation. Based on the results obtained the ability of nicotinyl-L-tyrosinate and nicotinyl-L-tryptophanate to act as cyto-protectors is concluded.

[Circulating immune complexes in families with positive history of ischemic stroke].

The concentrations and protein composition of immune complexes circulating in the blood of patients with residuals of ischemic stroke and their healthy relatives from families with positive history of stroke have been determined. The data obtained have been compared with the results of our previous study on determination of concentration and protein composition of immune complexes circulating in the blood of patients with acute ischemic stroke and healthy subjects. Basing on the results obtained we conclude that the elevated level of immune complexes in the blood of patients with residuals of stroke is not genetically determined but rather reflects alterations developing as the result of previous stroke. However, the protein composition of the immune complexes, particularly the presence of C-reactive protein, may, to a certain degree, reflect genetic predisposition to stroke.

[The role of cytokines in ischemic stroke].

The subjects of the study were patients with acute ischemic stroke (IS), patients with residual effects of IS, their healthy relatives in families with IS background, and healthy controls; blood levels of cytokines in these groups were compared. This included measurement of levels of tumor necrosis factor a, interleukins (IL-beta, IL-6), and chemokins (monocyte chemotoxic factor-1 and cytokine-inducible neutrophile chemoattractant.) The study presents experimental evidence of involvement of cytokines in molecular pathological mechanisms of generation and development of inflammatory immune response in patients with IS. The results show that therapeutic correction directed towards modulation of inflammatory immune response at the level of cytokine expression, is a necessary part of prevention and treatment of stroke, as well as successful rehabilitation of stroke patients.

[Concentration and protein composition of circulating immune complexes in the blood of patients with schizophrenia and subjects with positive familial history of disease].

A comparative study of blood serum concentrations and pathogenic properties of circulating immune complexes (CIC), along with identification of their protein composition, was conducted in 45 patients with schizophrenia, 15 their healthy relatives and 39 normal controls. In patients and their relatives, mean concentration of small CIC was within the normal range, while concentrations of giant, large and middle CIC were higher than those of the controls (p<0.001). Over 80% of schizophrenic patients and their relatives had pathogenic immune complexes in the circulation. Clinical and immunologic analysis of patients with schizophrenia revealed a correlation between the illness duration and CIC concentration for all sizes. Smokers had significantly lower levels of small CIC comparing to non-smokers. Determination of CIC composition in patients and relatives revealed a presence of specific proteins in the immune complexes with molecular weights of 36 and 25 kDa. The results suggest genetic determination of autoimmune processes in schizophrenia.

[Complement componenets, C3 and factors B, in the blood of patients with acute ischemic stroke].

Several studies suggest that activation of compliment cascade contributes to the development of the inflammatory immune response in ischemic stroke and results in tissue injury by initiating apoptosis and necrosis. It is proposed that suppression of the compliment activation may have positive effect on stroke severity and outcome. However, the majority of data relevant to involvement of the compliment in the pathogenethic mechanisms of stroke have been obtained in animal models of stroke that does not allow to extrapolate these data to humans. Our previous studies were the first ones demonstrated the involvement of both the classical and the alternative complement activation pathways in the pathogenetic mechanisms of generation and development of stroke in humans. In the present work, using immunoblotting, we determined the levels of key components of the classical and alternative pathways of complement activation, C3 and factor B, in the blood of patients with ischemic stroke. Comparing to healthy controls, patients with ischemic stroke are characterized by the increased level of C3 and decreased level of factor B.

[Prostaglandin D2-synthase from cattle brain: purification and physico-chemical properties]. / Prostaglandin D2-sintaza iz mozga byka: ochistka i fiziko-khimicheskie svoistva.

Prostaglandin D2-synthase has been purified from bovine brain for the first time and characterized. The enzyme has a molecular mass of 21 kDa and a pH optimum at 9.0 and is activated by dithiothreitol. The presence in the enzyme molecule of carbohydrates and lipids has been established and a preliminary conclusion made about the qualitative composition of the lipids. The ability of the enzyme to form aggregates under the influence of the detergent has been demonstrated. The role of the lipid component in the preservation of the enzyme monomeric structure as well as the possible mechanisms of action of the above inhibitors and activators are discussed.

[Purification of cytochrome B-561 from chromaffin granules and its physico-chemical properties]. / Ochistka tsitokhroma B-561 iz membran khromaffinnykh granul i ego fizikko-khimicheskie svoistva.

A soft method of purification of cytochrome-561 from the membranes of chromaffin granules has been developed. It permits isolating a protein in its natural microsurroundings, i.e. a complex with lipids, provided that a buffer with high ionic force is used without a detergent. This method helps obtaining an electrophoretically homogeneous preparation as a high-molecular lipoprotein hexamer whose molecular weight is about 400 kDa. Basic physicochemical parameters of this preparation (subunit composition, content and composition of lipids, heme content, spectra of optical absorption of the oxidized and reduced forms) are determined. Possible presence of two forms of cytochrome b-561 in the chromaffin granules is discussed.

[Acidic copper-containing proteins--an intermediate link to the electron transfer stage from cytochrome b-561 to dopamine-beta-monooxygenase]. / Kislyi med'soderzhashchii belok--promezhutochnoe zveno na stadii perenosa élektronov ot tsitokhroma b-561 k dofamina-beta-monooksigenaze.

The interaction of acidic copper-containing protein from the membranes of chromaffin granules has been investigated with cytochrome b-561 and dopamine-beta-monooxygenase from the same source. By the use of spectral and polarographic measurements it was demonstrated that the acidic copper-containing protein acts as an electron acceptor for cytochrome b-561 and as electron donor in the reactions, catalyzed by dopamine-beta-monooxygenase. According to the data obtained the possible function of the acidic copper-containing protein in vivo on the area of electron transfer chain between cytochrome b-561 and dopamine-beta-monooxygenase are discussed. The activation or inhibition of the electron transfer reactions by a variety of phospholipids, analogs of membrane lipids of chromaffin granules has been established. The experiments were performed in a model systems by the use of highly purified preparations of proteins and bilamellar liposomes and micelles, prepared from the corresponding phospholipids.