[The effects of preparation "Histochrome" in biochemical parameters of blood for patients with cardiopathologies].

The effects of the small doses of the preparation Gistochrome, containing natural polyhydroxynaphtoquinoine echinochrom A from flat sea urchin Scaphechinus mirabilis, on blood biochemical parameters have been studied in patients with cardiovascular diseases. Gistochrome administration influenced the LPO-antioxidant protection system, indicating reinforcement of antioxidant protection mechanisms. Gistochrome modulated the immune status and the plasma cytokine profile. Thus, Gistochrome may be recommended as means of additional therapy for patients with cardiovascular diseases for correcting the metabolic, immunological and redox impairments.

[Interaction of sea amemone Heteractis crispa Kunitz type polypeptides with pain vanilloid receptor TRPV1: in silico investigation].

Using methods of molecular biology we defined the structures of the 31 sea anemone Heteractis crispa genes encoding polypeptides which are structurally homologous to the Kunitz proteinase inhibitor family. Identified amino acid sequences have point residue substitutions, high degree of homology with sequences of known H. crispa Kunitz family members, and represent a combinatorial library of polypeptides. We generated their three-dimensional structures by homologous modeling methods. Analysis of their molecular electrostatic potential enabled us to divide given polypeptides into three clusters. One of them includes polypeptides APHC1, APHC2 and APHC3, which were earlier shown to possess a unique property of inhibiting of the pain vanilloid receptor TRPV1 in vitro and providing the analgesic effects in vivo in addition to their trypsin inhibitory activity. Molecular docking made possible establishing the spatial structure of the complexes, the nature of the polypeptides binding with TRPV1, as well as functionally important structural elements involved in the complex formation. Structural models have enabled us to propose a hypothesis contributing to understanding the APHC1-3 impact mechanism for the pain signals transduction by TRPV1: apparently, there is an increase of the receptor relaxation time resulted in binding of its two chains with the polypeptide molecule, which disrupt the functioning of the TRPV1 and leads to partial inhibition of signal transduction in electrophysiological experiments.

[Polypeptide toxin from sea anemone inhibiting proton-sensitive channel ASIC3].

Polypeptide toxin pi-AnmTX Hcr 1b-1 with a molecular weight 4537 Da was isolated from the whole body extract of sea anemone by a multistage liquid chromatography. The BLAST search algorithm revealed homology of the novel toxin amino acid sequence to the group of the known sea anemone toxins including BDS and APETx with similarity less then 50%. The toxin pi-AnmTX Hcr 1b-1 inhibited the amplitude of the fast component of integral ASIC3 current in electrophysiological studies on receptors expressed in Xenopus laevis oocytes. The calculated IC50 value was 5.5 +/- 1.0 microM. Among the known polypeptide toxins interacted with ASICs channels, the micro-AnmTX Hcr 1b-1 toxin is the least potent inhibitor that in our opinion correlates with a small amount of charged amino acid residues in its structure.

[Conformational stability of serine proteinase inhibitor from the sea anemone Heteractis crispa].

The influence of different environmental values of the pH and temperature on the spatial organization of serine proteinase inhibitor from the sea anemone Heteractis crispa (=Radianthus macrodactylus) on the level of tertiary and secondary structure was studied by CD spectroscopy. The molecule InhVJ was shown to possess a high conformational thermo- and pH-stability. We determined the point of conformational thermotransition of polypeptide (70 degrees C) after which the molecule gets denaturational stable state with conservation of 80% proteinase inhibitory activity. The significant partial reversible changes of molecule spatial organization were established to occur at the level of tertiary structure in the process of acid-base titration in the range of pH 11.0-13.0. This can be explained by of ionization of tyrosine residues. The molecule InhVJ is conformationally stable at the low pH values (2.0). The quenching of tyrosine residues by acrylamide showed that two of these residues are accessible to the quencher in full, while the third part is available.

[The comparative study of the medical action of lyuteolin, rosmarinic acid and echinochrom A at experimental stress-induced cardiopathology].

The method of the physical load in condition of the coronary circulation of the blood disturbance, caused mesaton injection, induced development rat cardiopathology, bring about of the heart function decompensation and 40% death of experimenthal animals. Under electronic-microscopic study of rat cardiomyocytes are discovered signs to disorganizations of mitochondrial apparatus of these cells. Administration to therapeutic mode of luteolin and echinochrome A preparations has provided to 100% animal probability of survival. At the same time, mitochondrial apparatus of cardiomyocytes was characterized by the normal parameter i.e. given preparations have provided of defensive adative effect at cardiomyocytes level. Similar activities for rosmarinic acid have not shown. Study some metabolic parameter and endocrine status animal has also allowed revealing of therapeutic effect of luteolin and echinochrome A. Findings be evidence of that echinochrome A and luteolin capable to play the important positive role in metabolism of cardiomyocytes by stimulating of mitochondrial biogenesis and by changing of adaptative mechanisms of the organism cardiovascular system protection.

[Investigation of luromarin efficacy on mice with experimental tick-borne encephalitis].

Oral use of Luromarin, extracted from sea algae Zostera asiatica, was efficient in protection of mice from lethal infection induced by highly pathogenic strain of TBE virus, by extending their average lifespan. Luromarin demonstrated potentiating action in combination with ribavirin and cycloferon.

[In vitro activity of luromarin against tick-borne encephalitis virus].

Luromarin, extracted from the sea alga Zostera asiatica, showed no cytotoxic activity when incubated with the SPEV cell culture and was characterized by virulicidal properties against highly pathogenic strain of TBE virus and the ability to inhibit the virus replication at the early stages of the infection.

[Effect of ginsenoside Rh2 on the development of transferred tumors and chemotherapy efficiency].

Experiments on C57B1/6 mice showed that ginsenoside Rh2 inhibited the growth and metastasis process of Lewis lung tumor and increased the antitumor and antimetastatic effects of cyclophosphamide. On the model of transferred melanoma B-16, ginsenoside Rh2 showed a tendency to increase the antiblastome effect of the cytostatic drug.

[A serine protease inhibitor from the anemone Radianthus macrodactylus: isolation and physicochemical characteristics].

A serine protease inhibitor with a molecular mass of 6106 +/- 2Da (designated as InhVJ) was isolated from the tropical anemone Radianthus macrodactylus by a combination of liquid chromatography methods. The molecule of InhVJ consists of 57 amino acid residues, has three disulfide bonds, and contains no Met or Trp residues. The N-terminal amino acid sequence of the inhibitor (19 aa residues) was established. It was shown that this fragment has a high degree of homology with the N-terminal amino acid sequences of serine protease inhibitors from other anemone species, reptiles, and mammals. The spatial organization of the inhibitor at the levels of tertiary and secondary structures was studied by the methods of UV and CD spectroscopy. The specific and molar absorption coefficients of InhVJ were determined. The percentage of canonical secondary structure elements in the polypeptide was calculated. The inhibitor has a highly ordered tertiary structure and belongs to mixed alpha/beta or alpha + beta polypeptides. It was established that InhVJ is highly specific toward trypsin (Ki 2.49 x 10(-9) M) and alpha-chymotrypsin (Ki 2.17 x 10(-8) M) and does not inhibit other proteases, such as thrombin, kallikrein, and papain. The inhibitor InhVJ was assigned to the family of the Kunitz inhibitor according to its physicochemical properties.

[Interaction investigation of trypsin inhibitor from sea anemone Radianthus macrodactylus with proteases].

The interaction of inhibitor VJ (InhVJ), isolated from sea anemone R. macrodactylus, with different proteases was investigated. The following enzymes were tested: serine proteases (trypsin, alpha-chymotrypsin, plasmin, thrombin, kallikrein), cysteine protease (papain) and aspartic protease (pepsin). Inhibitor VJ interacted only with trypsin and 6-chymotrypsin. Kinetic and thermodynamics parameters of intermolecular complexes formation were determined: KD = 7,38 x 10(-8) M and 9,93 x 10(-7) M for pairs InhVJ/trypsin and InhVJ/alpha-chymotrypsin, respectively.

[Primary structures of actinoporins from sea anemone Oulactis orientalis].

Partial amino acid sequences of the actinoporins Or-A (136 aa) and Or-G (144 aa) isolated from the Sea of Japan sea anemone Oulactis orientalis were determined by sequencing the clones obtained by the amplification of genomic DNA and cDNA with specific primers to the N-terminal regions of the 0. orientalis actinoporin sequences and to the C-terminal region, which is known to be highly conservative in all the known actinoporin sequences. The complete structures of Or-A (165 aa) and Or-G (173 aa) were established by sequencing the cDNA clones obtained by the fast amplification of 3'-ends of cDNA. A comparative analysis of the amino acid sequences of the Oulactis actinoporins with those of actinoporins from tropical species revealed considerable differences in the structures of their N-terminal fragments and their membrane-binding sites. We believe that these differences could explain lower hemolytic activities of Or-A and Or-G than that of actinoporins from the tropical species.

[Hepatoprotective properties of balm Herbamarin and hydrolysates from marine invertebrates in toxic hepatitis and ethanol intoxication].

Protective properties of a syrup balm "Herbamarin" and food hydrolysates of scallop, octopus and crab were investigated using experimental toxic hepatitis and ethanol intoxication. Preventive administration of the balm and hydrolysates to animals subjected to an intoxications by 40% alcohol and CCl4 normalized clinical-diagnostic parameters of liver and blood plasma of experimental animals.

[Actinoporins from the Sea of Japan anemone Oulactis orientalis: isolation and partial characterization].

Two cytolytic toxins (cytolysins Or-A and Or-G) were isolated from the Sea of Japan anemone Oulactis orientalis and characterized. Their purification scheme involved a hydrophobic chromatography on Polychrom 1, a gel filtration on Akrilex P-4, a cation-exchange chromatography on CM-32 cellulose, and a reversed-phase HPLC on a Nucleosil C18 column. The molecular masses of Or-A and Or-G were determined by SDS-PAGE in 14% PAG to be ca. 18 kDa. The absence of Cys residues and a high content of basic amino acid residues are characteristic of their amino acid compositions. The hemolytic activities of Or-A and Or-G were found to be 295.86 and 322.58 HU/mg, respectively; these are by three orders of magnitude lower than those of sphingomyelin-inhibitable cytolysins from the tropic sea anemones. The amino acid sequences of the N-terminal fragments of Or-A and Or-G were determined to be ATFRVLAK and GAIIAGAA, respectively. Action of the cytolysins on the erythrocyte membrane is inhibited by exogenous sphingomyelin. They form ion channels in bilayer lipid membranes with the conductivity of 16, 32, and 40 pSm in 0.1 M NaCl and 168, 240, and 320 pSm in 1 M NaCl at pH 7.2. Therefore, they were attributed to the group of actinoporins.

[Effects of Gerbamarin T balms on functions of the cardiovascular and hepatobiliary systems]. / Vliianie bal'zamov "Gerbamarin" T na funktsional'noe sostoianie serdechno-sosudistoi i gepato-biliarnoi sistem.

The effects two balms of "Herbamarin" T series on some parameters of cardiovascular and hepatobiliary systems were studied in mice either kept on the cholesterol diet or treated with CCl4. The results allow to recommend these balms as additional components to the therapy of various cardiovascular and hepatic diseases.

[Effect of biologically active substances from marine organisms on lipid peroxidation-antioxidant defence system in gastroduodenitis, associated with hyperlipidemia]. / Sostoianie sistemy perekisnoe okislenie lipidov--antioksidantnaia zashchita pri gastroduodenite, assotsiirovannom s giperlipidemiei, pod deistviem biologicheski aktivnykh veshchestv morskikh gidrobiontov.

An experimental model of gastroduodenitis combined with hyperlipidemia was used to study the effects of the product Zolotoi rog (Golden Horn) which is a composition of biologically active substances of marine organisms and honey. It was found that a course administration of Zolotoi rog in a dose 2.5 mg/kg b.w. improves histomorphology of gastric mucosa, acts hypolipidemically, raises reserves of the antioxidant system of the body and suppresses intensity of lipid peroxidation.

[A specialized product with antioxidant action in the treatment of lipid disorders in experimental hyperlipoproteinemia]. / Spetsializirovannyi produkt antioksidantnogo deistviia v korrektsii lipidnykh narushenii pri éksperimetal'noi giperlipoproteidemii.

At an experimental model of alimentary hyperlipoproteinemia the medical effect of specialized product "Zolotoi Rog", a composition of biologically active substances, antioxidants and honey, isolated from marine organisms, was analysed. The hypolipidemic effect of this product was defined at violations of lipid metabolism of blood and liver of animals. Deterioration of the processes of lipids peroxidation and raising activity of antioxidant system of an organism, were revealed.

[Isolation and characteristics of high molecular weight cytolysins from the sea anemone Radianthus macrodactylus]. / Vydelenie i kharakteristika vysokomolekuliarnykh tsitolizinov morskoi aktinii Radianthus macrodactylus.

Three cytolytic toxins (RTX: RTX-A, RTX-S, and RTX-G) were isolated from the sea anemone Radianthus macrodactylus and characterized. The purification scheme involved hydrophobic chromatography on Polychrom-1, batch-chromatography on CM-23 cellulose, gel filtration on Akrilex P-4, cation-exchange chromatography on CM-32 cellulose, and HPLC on an ion-exchange Ultropac TSK CM-3SW column and a reversed-phase Silasorb C18 column. The molecular masses of RTXs (ca. 20 kDa) were determined by SDS-PAGE in a density gradient of PAG. They are highly basic polypeptides (pI of 9.8 for RTX-A and RTX-S and 10.5 for RTX-G) containing similar amino acid compositions with a high content of basic and hydrophobic residues and the absence of Cys residues. The hemolytic activities of RTX-A, RTX-S, and RTX-G were determined to be 3.5, 5.0, and 1.0 x 10(4) HU/mg, respectively. Exogenous sphingomyelin inhibits their action on the erythrocyte membrane. The N-terminal sequence of RTX-A was determined to be ALAGAIIAGAGL/KGLKI/FLIEVLGEG--V/NKVKI-.

[Low-molecular cytolysins and trypsin inhibitors from sea anemone Radianthus macrodactylus. Isolation and partial characterization]. / Nizkomolekuliarnye tsitoliziny i ingibitory tripsina iz morskoi aktinii Radianthus macrodactylus. Vydelenie i chastichnaia kharakteristika.

Two low-molecular cytolytic toxins (RmI and RmII) and four trypsin inhibitors were isolated from the aqueous extract of sea anemone Radianthus macrodactylus. The method of isolation involved precipitation with acetone, gel filtration on acrylex P-4, ion-exchange chromatography on CM-32 cellulose, affinity chromatography on trypsin-binding sepharose 4B, ion exchange chromatography on an Ultrapore TSK CM-3SW column, and reversed phase HPLC on a Silasorb C18 column. RmI, RmII, and JnI inhibitor displayed molecular masses 5100, 6100, and 7100 Da, respectively, when subjected to SDS-PAGE. The isoelectric points were 9.2 and 9.3 for RmI and RmII, respectively. The amino acid composition and N-terminal amino acid residue (glycine) were determined for RmI, RmII, and JnI. Both proteins were nontoxic to mice and crabs. Hemolytic activity was determined to be 25 and 20 HU/mg for RmI and RmII, respectively, and their action on erythrocyte membrane was not inhibited by exogenous sphingomyelin. RmI and RmII exhibited antihistamine activity.

[The effect of chemical modification on the immunochemical activity of Rm-III neurotoxin from the anemone Radiantus macrodactylus]. / Vliianie khimicheskoi modifikatsii na immunokhimicheskuiu aktivnost' neirotoksina Rm-III iz aktinii Radiantus macrodactylus.

A chemical modification was used for studying the organization of antigenic determinants of neurotoxin Rm-III from the sea anemone Radianthus macrodactylus. Immunochemical experiments were performed using competitive enzyme-linked immunosorbent assay (ELISA) with polyclonal antibodies to Rm-III. The modification affected N-terminal amino group of Gly1, Lys4, Arg13, Trp30 residues, a residue in the Lys46-Lys47-Lys48 sequence, two different residues in the Asp6-Asp7-Glu8 sequence in two samples of the toxin, and two disulphide bonds. Only the modification of the disulphide bonds led to a considerable change in the toxin's affinity to antibodies. The modification of Trp30 resulted in two-fold decrease of the toxin concentration necessary for 50% inhibition of the test-system, whereas upon modification of any other amino acid residue this concentration increased but not more than by 2.2 times. It is suggested that Rm-III sequence lacks individual residues which are of great importance for the toxin's antigenic activity, its conformation being of vital importance for the formation of the toxin's antigenic determinants.

[Immunochemical study of neurotoxins from Radianthus macrodactylus actinin]. / Immunokhimicheskoe izuchenie neirotoksinov iz aktinin Radianthus macrolus.

Polyclonal antibodies to neurotoxin Rm-III from sea anemone Radianthus macrodactylus have been obtained. Constants of inhibition of the Rm-III binding to its antibodies by the homologous toxins have been determined. Antigenic activity of the second type toxins is shown to depend not only on the degree of homology but also no the type of substitution in the amino acid sequence. As shown by the calculation methods, the antigenic determinants of all the homologues have similar positions. Amino acid residues at positions 2, 11, 20, 28, and 46-48 seem to be included into the antigenic sites of the toxins studied.