[Transthyretin gene V30M, H90N, and del9 mutations in cardiomyopathy patients from St. Petersburg].

A search of transthyretin (TTP) gene mutations was conducted in patients with cardiomyopathies from St. Petersburg. Mutations H90N, V30M, G47A, and deletion (del9) of nucleotides GACTTCTCC in position 6776 from the start codon of the TTP gene (in position 98782 according to reference sequence AC079096 (NCBI) was found.

[Characteristics of rat ceruloplasmin from the serum of animals, which received salts of silver with food].

Abiogenic Ag(I) ions have electronic structure, similar to Cu(I) ions and can compete with Cu(I) for binding sites of proteins which transport copper from extracellular media to sites of cuproenzyme formation in the cell. Rodents receiving Ag-salts with food develop extracellular deficiency of copper associated with ceruloplasmin (Cp, the major copper-transporting protein in blood serum of vertebrates). The present work focuses on the studies of biochemical and physicochemical properties of Cp, obtained from blood serum of rats, which received AgCl with food for 4 weeks (Ag-rats).

Effect of a deficiency of ceruloplasmin copper in blood plasma on copper metabolism in the brain.

Copper deficiency in adult rats was induced by addition of silver chloride to the feed. The concentrations of silver, copper, iron, and zinc and relative activity of genes for copper transporting proteins and copper enzymes were measured in the cortex, cerebellum, hippocampus, amygdala, pituitary gland, and hypothalamus. Silver was accumulated only in the hypothalamic-pituitary system.

Changes in copper metabolism in different compartments of the brain in rats with induced fibrillogenesis.

Fibrillogenesis was induced in rats by injection of a fragment of neurotoxic protein, beta-amyloid protein precursor, into the cerebral ventricle. Copper, iron, and zinc concentrations and relative activities of genes of copper-transporting protein and extracellular and intracellular cuproenzymes were evaluated in different brain compartments of these animals. Copper and zinc concentrations decreased significantly in different compartments of the brain of rats with experimental fibrillogenesis, while iron content did not change.

[The study of preimplantation development of mice embryos labelled with green fluorescent protein gene (EGFp)].

One of the crucial problems of developmental biology is the study of mechanisms of regulation of gene expression in early embryogenesis. Here we studied dynamics of mosaic appearance of a marker fluorescent protein in in vitro developing mice embryo derived from zygotes after microinjections to male pronuclei of cloned DNA fragment carrying EGFP under control of different promoters. Main attention was paid to initial stages of development, when structural rearrangements and reprogramming of both parental genomes, activation of zygotic genes, and control of development by embryo genome take place.

[Expression of recombinant actin 5C from Drosophila in the methylotrophic yeast Pichia pastoris].

A system for actin expression in cells of yeast Pichia pastoris was constructed. Drosophila actin 5C, by 90% homologous to beta-actin of higher eukaryotes, was used as a target protein. To improve the procedures of target protein biosynthesis in yeast cells and of extraction and purification of recombinant actin the fusion protein GFP-actin 5C, having fluorescence protein GFP as a reporter part, was expressed and purified.

[The role of quarternary structure in fluorescent protein stability].

The stability of fluorescent proteins (FPs) is of great importance for their use as reporters in studies of gene expression, protein dynamics and localization in cell. A comparative analysis of conformational stability of fluorescent proteins, having different association state was done. The list of studied proteins includes EGFP (monomer of green fluorescent protein, GFP), zFP506 (tetramer GFP), mRFP1 and "dimer2" (monomer and dimmer of red fluorescent protein), DsRed1 (red tetramer).

[Room temperature phosphorescence of amorphous aggregates and amyloid fibrils resulting from protein misfolding].

Using actin, alpha-lactalbumin and insulin as examples, it was shown that the formation of amorphous aggregates of proteins and amyloid fibrils leads to an increase in the rigidity of tryprophan and tyrosine residues micro-environment and, consequently, to the appearance of tryptophan (tyrosine) room temperature phosphorescence (RTP). RTP was used for examining a slow intramolecular mobility of native (G-, F-form) and inactivated (I) rabbit skeletal muscle actin during the process of GdnHCl induced protein unfolding. This method made it possible to confirm that an essentially unfolded intermediate precedes the formation of inactivated actin.

[Isolation of stable human ceruloplasmin and its interaction with salmon protamine].

An interaction was discovered between ceruloplasmin (CP, a ferro-O2-oxidoreductase, EC 1.16.3.

[Kinetics of inactivation of lectin: analysis of a method of tryptophan phosphorescence at room temperature].

The internal dynamics of muscle actin during inactivation induced by guanidine hydrochloride (0.5-1.8 M) was studied by the method of room-temperature tryptophan phosphorescence (RTTP).

[Suppression of nonsense mutations in the Dystrophin gene by a suppressor tRNA gene].

Nonsense mutations in the dystrophin gene are the cause of Duchenne muscular dystrophy (DMD) in 10-15% of patients. In such an event, one approach to gene therapy for DMD is the use of suppressor tRNAs to overcome the premature termination of translation of the mutant mRNA. We have carried out cotransfection of the HeLa cell culture with constructs containing a suptRNA gene (pcDNA3suptRNA) and a marker LacZ gene (pNTLacZhis) using their polymer VSST-525 complexes.

[Tryptophan phosphorescence of nascent and inactivated actin at the room temperature].

Millisecond internal dynamics of native and inactivated actin from rabbit skeletal muscle was examined using room temperature phosphorescence. Inactivated actin was prepared by incubation of G-actin at 70 degrees C, by treatment with 4 M urea or 1.5 M guanidinium hydrochloride, renaturation from fully unfolded state or by Ca2+ ion removal.

[Interaction of lactoferrin with plasmid DNA studied by laser correlation spectroscopy].

The interactions of lactoferrin with plasmid DNAs substantially differing in the number of affine regions were studied. The dissociation constants for protein-nucleic acid complexes were determined. The possibility of standardizing the conditions for preparing multicomponent systems for gene-substituting therapy by laser correlation spectroscopy is discussed.

[Expression of marker genes in muscle fibers after transfection in vivo, mediated by lactoferrin].

The capacity of milk iron-transporting human protein lactoferrin (LF) to deliver genetic constructions into cells was studied in an effort to correct hereditary defects. The purified LF and LF conjugates containing either polylysine (C-1) or both polylysine and ficoll (C-2) were bound to plasmid DNA. These complexes were injected into mouse muscles, and the expression of the marker genes was tested immunochemically.

[Spectrofluorometric analysis of protein fractions of blood serum of health donors and patients with kidney diseases].

A comparison was made of ultra-violet fluorescence characteristics of albumin enriched protein fractions of serum blood of 35 glomerulonephritis (GN) patients, 30 uremic haemodialysis patients, and 40 donors. It has been found that fluorescence spectra of the albumin enriched fractions of serum from GN patients are blue shifted as compared with those of donors' serum preparations. At the same time fluorescence spectra of the albumin enriched fractions of donors' serum are similar, while those of GN patients vary significantly.

[Analysis of molecular microheterogeneity of plasma glycoproteins in a patient with sialidosis 1].

Using specific polyclonal monovalent antibodies, the molecular microheterogeneity of acute phase proteins: orosomucoid, alpha1-antitrypsin and ceruloplasmin, circulating in peripheral blood of a healthy donor and a patient with a hereditary deficiency of lysosomal neuraminidase (syalidosis I or the cherry stone syndrome), was analyzed by use of 2D electrophoresis. The specific distinctions due to a deficiency [caused by a deficiency] of lysosomal neuraminidase were revealed in the population of ceruloplasmin molecules, but not in the molecules of alpha1-antitrypsin and orosomucoid (alpha 1-acid glycoprotein). The molecular genetic bases of molecular microheterogeneity of some plasma glycoproteins and the possible use of natural models in studies of GP's functional role and the pathways of its transfer after internalization into non-hepatocytic (?) cells are discussed.

[Influence of ceruloplasmin on the embryotoxic effect of silver ions].

The effect of alimentary administration of silver salts upon embryogenesis in rats has been studied. Feeding of AgCl to pregnant female rats throughout gestation did not result in any alterations in their physiological functions, although the active copper-containing ceruloplasmin (Cp) was eliminated from the blood stream. However, anomalous development of embryos, their prenatal death or total mortality of newborn rats within the first 24 hours after birth were evidenced.

[The cytophotometric determination of the transferrin content of rat hepatocytes by using monoclonal antibodies].

Monoclonal antibodies specifically recognizing rat transferrin were produced. Optimal conditions of fixation and carrying out immunocytochemical reaction were determined. A combined method is proposed based on the indirect immunoperoxidase reaction and Feulgen procedure to be used for studying transferrin contents in hepatocytes with different ploidy.

[Various properties of the ceruloplasmin receptor, isolated from human erythrocyte membranes].

An electrophoretically pure preparation of ceruloplasmin (CP) receptor which retains its ability to bind to CP was isolated from human erythrocyte membranes. It was found that in terms of molecular mass, number and size of spontaneously proteolytic fragments as well as antigenicity, the CP receptor molecule strongly resembles that of CP. A comparative analysis of two-dimensional peptide maps of full tryptic digests of the both protein revealed that about 30% of CP peptides are identical in respect of their electrophoretic and chromatographic mobilities which points to the genetic independence of these proteins.

[Properties of silver-containing rat ceruloplasmin].

Ceruloplasmin (Cp) was isolated from the sera of albino rats fed with silver nitrate (60 mg/kg of body weight). The oxidase activity of the enzyme was sharply decreased, while its concentration in the blood (as assayed immunologically) was slightly lower than in controls. The drop in the oxidase activity was caused by the replacement of several coppers by silver ions in the Cp molecule.

[Etiologic significance of hereditary deficiency of proteinase alpha1-inhibitor in the formation of respiratory diseases].

A study was made of the content of protease alpha 1-inhibitor and of the phenotyping of protease alpha 1-inhibitor subtypes in 666 patients with different chronic non-specific pulmonary diseases. It is concluded that pronounced deficiency of protease alpha 1-inhibitor is of importance in the formation of primary emphysema, chronic obstructive bronchitis and bronchial asthma. The clinical characteristics of obstructive pulmonary diseases marked by protease alpha 1-inhibitor deficiency have been investigated.

[Synthesis and secretion of ceruloplasmin by isolated rat hepatocytes].

The synthesis and secretion of ceruloplasmin (Cp) by isolated rat hepatocytes were investigated. Cp released by liver cells appeared to have properties similar to those of the blood-circulating protein, i. e.