[Amyloid-like fibrils forming and fibroblasts destruction in Tenon's capsule in progressive myopia as a result of pigment epithelium derived factor resistance to restricted proteolysis].

We have shown previously the presence of full length (50 kD) and truncated proteolytic form (45 kD) of pigment epithelium derived factor (PEDF) in the eye Tenon's capsule in progressive myopia. The full length PEDF is prevalent in myopia that correlates with breach in collagen fibrils forming. Immunohistochemical analysis of Tenon's capsule with polyclonal antibodies to PEDF revealed PEDF in control group being exclusively inside fibroblasts, whereas in myopia, PEDF was distributed extracellularly as halo around blasted fibroblasts. By means of atomic force microscopy and immunodot analysis with anti amyloid fibrils antibodies the ability was studied of recombinant PEDF fragments to form fibrils. Only full length PEDF was shown to form amyloid like fibril structures, but not the truncated form. Accumulation offibrils results in fibroblasts destruction and might be the cause of changes in biochemical and morphological structure of Tenon's capsule observed in myopia.

[Identification in the rat olfactory epithelium new subgroup YM-1 chitinase-like protein].

Novel protein with a molecular mass of ~43 kDa from rat olfactory epithelium in pathophysiological conditions was discovered. Its amino acid sequence and affiliation with the family 18 glycohydrolase subgroup of chitinase-like proteins YM-1 were determined.

The novel approach to the protein design: active truncated forms of human 1-CYS peroxiredoxin.

The object of the present study is the verification of a new approach to the design of the active truncated forms of enzymes. The method is based on a new way of investigating the protein sequences--the ANalysis of Informational Structure (ANIS). The analysis of informational structure allows to determine the hierarchically organized structures (IDIC-trees) formed by the sites with the Increased Degree of Informational Coordination between residues. The proposed approach involves the consequent removal of the fragments corresponding to the individual IDIC-trees from the wild-type enzyme sequences. The described procedure was applied to the design of the active truncated form of human 1-CYS peroxiredoxin (PrxVI). Two variants of the PrxVI truncated sequences were proposed according to ANIS method. These truncated forms of the enzyme were expressed in E. coli and purified. The respective antioxidant activities were measured. It was shown that one of the truncated recombinant proteins retains more than 90% of the wild-type PrxVI enzymatic activity. According to the results of our study we can assume that ANIS method can be an effective tool for the design of the active truncated forms of the enzymes or the chimeric proteins which combine the enzymatic activities of their wild-type prototypes.

[Identification and expression of haponin, a new protein from HL-60 cells].

We found a new protein haponin (an HLDF-alike protein) in promyelocyte HL-60 cells that is immunoreactive to polyclonal antibodies against HLDFbeta. Determination of the partial primary structure of the protein allowed us to reveal an immunogenic peptide of haponin and, on the basis of the amino acid sequence of this peptide, the degenerate primers were synthesized, which enabled us to clone the full-size cDNA of haponin. The stable heterologous expression of this cDNA in E. coli cells (Rosetta strain) was obtained. Preparations of natural and recombinant proteins exhibited antigenic cross-reactivity to polyclonal antibodies against this peptide.

[Binding of phosphatidylinositol-3,4,5-triphosphate to 45-kDa sec14-like protein from the rat olfactory epithelium in liposomes].

The binding of phosphatidylinositol-3,4,5-triphosphate to a protein with molecular mass of 45 kDa from rat olfactory epithelium (p45) was investigated using a model membrane system. Liposomes containing a mixture of phospholipids (phosphatidylethanolamine, phosphatidylcholine, and phosphatidylinositol-3,4,5-triphosphate) were used in the study. The binding of the protein to liposomes caused by its interaction with phosphatidylinositol-3,4,5-triphosphate was confirmed by cosedimentation and immunoblotting with chemiluminescent detection using monoclonal antibodies to the native protein p45. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2006, vol. 32, no. 3; see also http://www.maik.ru.

[A proteomic approach to study of the function of the Sec14p-like p45 protein].

We undertook a search for proteins interacting with protein p45 by the method of two-hybrid screening in order to determine the function of the Sec14p-like protein p45. A screening of the yeast library of rat lung cDNA, six proteins specifically activating the reporter genes of a two-hybrid system and 21 unlikely protein partners of protein p45 were identified. The most likely candidate for the role of a p45 partner is the surfactant protein C (Sftpc). These results and previous studies led us to the hypothesis that protein p45 fulfills its protective function by participating in the biogenesis of cell membranes. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 3; see also http://www.maik.ru.

Functional expression and properties of Sec14p-like protein with molecular mass 45 kD from rat olfactory epithelium.

cDNA of Sec14p-like water-soluble protein with molecular mass 45 kD from rat olfactory epithelium was expressed in Escherichia coli Rosetta cells. The expression product was purified by a two-step chromatographic procedure on DEAE-Sepharose and Sephacryl S-200. The identity of structural and functional characteristics of the recombinant and native proteins was demonstrated by CD, mass spectrometry, and Western blotting. Using several lipids immobilized on nitrocellulose membranes, it was shown that phosphatidylinositol-3,4,5-triphosphate is the specific ligand for the studied protein.

[Effect of allotransplantation of embryonic pluripotent progenitor cells on dynamics of systemic arterial blood pressure in spontaneously hypertensive rats]. / Vplyv alotransplantatsiï embrional'nykh pliurypotentnykh prohenitornykh klityn na dynamiku systemnoho arterial'noho tysku u shchuriv zi spontannoiu hipertenziieiu.

The influence of embryonic pluripotent progenitor cells on the dynamics of the systemic arterial pressure was examined in rats with genetically determined arterial hypertension. It has been established that the single intravenous administration of the embryonic pluripotent and progenitor cells to spontaneously hypertensive rats in the amount of 5 x 10(7) ml lead to the lowering of the systemic arterial pressure for one month.

Recombinant human peroxiredoxin VI: preparation and protective properties in vitro.

cDNA of human peroxiredoxin VI, one of the recently discovered novel antioxidant proteins, was expressed in Escherichia coli cells. The expression product was obtained in water-soluble form and purified by a two-step chromatographic procedure using DEAE-Sepharose and Sephacryl S-200. According to CD data, the polypeptide chain of the recombinant human peroxiredoxin VI contains approximately 40% alpha-helical region and 30% beta-structure, which is the same as for native rat peroxiredoxin VI. The protective properties of the recombinant protein determined as its ability to prevent the inactivation of glutamine synthetase from E. coli in a model oxidation system were comparable with the protective properties of native rat peroxiredoxin VI.

[Effect of irbesartan, an antagonist of AT-1 receptors for angiotensin II, on L-arginine metabolism in arterial hypertension]. / Diia irbezartanu--antahonista AT1-retseptoriv anhiotenzynu II na obmin arhininu za umov arterial'noï hipertenziï.

Effects of an antagonist of AT-1 receptors for angiotensin-II (Ang-II) irbezantane on the NO-synthase and arginase ways of the metabolism of L-arginine were studied in plasma and erythrocytes of the patients with arterial hypertension. The intensity of the non-oxidative arginase way of L-arginine metabolism in plasma and erythrocytes has been shown to be inhanced at hypertension versus the normotensive patients, while the activity of the alternative oxidative NO-synthase way was reduced. Inhibiting AT-1 receptors for Ang-II with high-affinity antagonist irbezantane normalized the ratio between two alternative ways of L-arginine metabolism through inhibiting the arginase way and reciprocal activating the NO-synthase way both in human plasma and erythrocytes.

[Effect of irbesartan--angiotensin II type I receptor inhibitor on oxidative metabolism of lipids in essential hypertension]. / Diia irbezartanu--prolonhovanoho antahonista AT1-retseptoriv anhiotenzynu II na okysnyi metabolizm lipidiv za umov hipertonichnoï khvoroby.

We evaluated the alters in plasma vasoconstriction eicosanoids (LTC4, TXB2) and diene conjugates (DC) levels in patients with essential hypertension (EH) after chronic (30 days, 235 mg per day) of irbezartane (inhibitor of ANG II type 1 receptor with prolongation action "Aprovel" from "Sanofi") oral administration. Patients with EH have significantly higher plasma both LTC4, TXB2 and DC levels then healthy controls. This imbalance can be beneficially modulated by chronic irbezartane ("Aprovel") administration. It is concluded that ANG II type 1 receptors can be involved in regulation of free arachidomc acid oxidation.

[The effect of the long-term treatment with calcium antagonists on myocardial function in patients with hypertension]. / Vliianie dlitel'nogo lecheniia antagonistami kal'tsiia na funktsional'noe sostoianie miokarda u bol'nykh gipertonicheskoi bolezn'iu.

Prolonged effect (6-12 months) of verapamil and nifedipine on the systolic and diastolic function of the myocardium was studied in 48 patients with hypertensive disease (stage II). It was established that in patients with a high ejection fraction (55% and more) the two drugs produce a distinct reduction of the pumping heart function. The two drugs increase the rate of early diastolic filling. They also may reduce the arterial pressure via different hemodynamic mechanisms: vasodilatation or via reduction of cardiac output.

[A comparative analysis of the antihypertensive activity of calcium antagonists and adrenoblockers in long-term treatment]. / Sravnitel'nyi analiz antigipertenzivnoi aktivnosti antagonistov kal'tsiia i adrenoblokatorov pri dlitel'nom lechenii.

Essential hypertension stage II was treated by calcium antagonists, alpha- and beta-adrenoblockers in 362 patients. The drugs were compared for hypotensive efficiency at rest and exercise. A significant hypotensive effect was achieved in 81%, 58%, 44%, 43% and 37% of patients treated with labetalol, nifedipine, nadolol, propranolol, diltiazem, respectively. An increase in verapamil daily dose from 240 to 600 mg led to a rise in efficacy from 27 to 75% without an increase in the number of side effects. The antihypertensive effect of the drugs is shown to persist in long-term treatment as shown by exercise tests and static loads.