The use of filamentous phage M13 in protein engineering

M13B1 vector based on the filamentous phage M13 has been constructed. M13B1 phage carries the gene of resistance to ampicillin and contains the unique site of recognition for BamHI restriction endonuclease in gene VIII coding for the major coat protein. BamHI restriction site has been inserted into the gene of the major coat protein by means of oligonucleotide directed mutagenesis. The synthetic DNA fragment coding for the model peptides has been inserted through BamHI site into the M13B1 DNA. The possibility of inserting foreign peptides into the N-terminus at maintaining the viability of hybrid phages has been shown. The differences in specificity of the recombinant phage maturation have been determined by analysing the amino acid sequence of B-protein.     

Directed reconstruction of the influenza virus hemagglutinin gene

A new approach to create chimeric genes by directed exchange of oligonucleotide fragments was developed. By oligonucleotide-directed mutagenesis a few deletion mutants of the influenza virus hemagglutinin (HA) gene were obtained. These variants of HA gene contain unique restriction sites in DNA regions coding for the A and B epitopes of the HA molecule. The obtained special vectors may be used for cloning DNA fragments coding for new amino acid sequences in internal sites of the HA gene.     

Electroencephalographic analysis of inter-central relations in hypothalamo-reticulo-limbic brain structures during treatment with corticosteroids and neuropeptides

Intercentral relations between hypothalamus, limbic system and reticular formation were studied in rabbits and rats under systemic and central action of DSIP, ACTH, corticosteroids and stress (aggressive-defensive behaviour). The results obtained demonstrate changes in the adrenal cortex resulting from stress-inducing adrenocortical hormone content. The increase was achieved by the rise in ACTH level resulting in corticosteroid level elevation (endogenous elevation-aggressive behaviour) and by corticosteroid injections (exogenous elevation). Correlation analysis of structural interrelations after ACTH and corticosteroid injections demonstrated an increased correlation between hypothalamo-reticular-limbic structures. DSIP was shown to have an opposite effect. Correlation analysis revealed the potentials for the formation of new functional interrelations between hypothalamo-reticular-limbic structure in the motivation of aggression (stress) and the levels of corticosterone and DSIP. DSIP action depends on the initial corticosteroid blood level and is more marked in stress-inducing concentrations.     

Construction of original artificial immunogens--candidate vaccines against HIV-1

An original method for making effective artificial vaccines has been developed. Immunogens are virus-like particles containing a DNA molecule covered with recombinant proteins carrying the pathogen epitopes. The recombinant proteins are exposed on the surface of the particle and are attached to DNA via spermidine-polygluquine-glutathione or galactopyranoside conjugates (depending on the hybrid protein composition). Two variants of artificial immunogens-candidates for antiHIV vaccine have been prepared.     

Use of the luciferase reporter constructs for investigation of the capacity of Noggin2 protein to inhibit cell signaling pathways in Xenopus laevis embryos

Noggin (Noggin1) protein inhibits Smad1-dependent TGF-?? (BMP) signaling cascades by extracellular binding of BMP proteins. Recently, we identified two previously unknown members of the Noggin family, Noggin2 and Noggin4 proteins. In this work, using luciferase reporter constructs, it was shown that Noggin2 is able to inhibit in addition to the BMP-signaling cascade, the Activin/Nodal and Wnt signaling pathways in Xenopus laevis early embryos. The inhibition efficiency of Noggin2 is comparable to well-known extracellular inhibitors Cerberus and Follistatin. In addition to revealing novel properties of Noggin2, this work demonstrates that luciferase reporter constructs are a convenient tool for studying the regulation of molecular signaling cascades in the model of Xenopus laevis embryos.     

The interaction of secreted proteins Noggin4 and Wnt8 from <Emphasis Type="Italic">Xenopus laevis</Emphasis> embryos

We demonstrated that the secreted protein Noggin4 from Xenopus laevis was capable of the in vitro binding to the secreted factor Wnt8, one of the ligands of the Wnt/betaCatenin signaling pathway. It was also shown that posttranslational modifications occurring during secretion of these proteins from the embryonic cells were necessary for their effective interaction. Also, we proposed a method for the preparation of physiologically active secreted morphogenic proteins from the intercellular space of the Xenopus laevis embryos.     

The specific features of coronary artery lesion in patients with signs of high cardiac risk in non-ST segment elevation acute coronary syndrome

The paper covers risk stratification for cardiac events in patients with non-ST segment elevation acute coronary syndrome. It defines an association between risk factors, the specific features of coronary artery lesions, and the outcomes of the disease in relation to the used treatment policy. Particular emphasis is laid on the factors that are a handicap to the detection of a symptom-associated coronary artery in this category of patients.