[Comparative cytogenetic study of the tetraploid Matricaria chamomilla L. and Matricaria inodora L].

A comparative cytogenetic study of the autotetraploid breed of Matricaria chamomilla L. (M. recutita L.) and Matricaria inodora L. was carried out by DAPI-banding, fluorescent hybridization in situ (FISH) with 26S and 5S rDNA probes, and analysis of meiosis. All chromosomes were identified in both karyotypeson the basis of DAPI-banding images and 26S and 5S rDNA distribution, and species-specific idiograms were composed for both M. chamomilla and M. indora taking into account the polymorphous variants of DAPI-banding images, showing the location of the 26S and 5S rDNA sites.

[Scientific approaches to development of medicinal formulation based on biotechnological substance].

The study demonstrated possible design of a medicinal formulation in the form of suppositories comprising human recombinant interferon-alpha2 and dry aloe extract. The approaches to the development of the suppositories were technology-derived. No interaction between the active and auxiliary components was proved by solid state 1H-NMR spectroscopy. The specific activity of the drug was investigated.

[Comparative cytogenetic study of the forms of Macleaya cordata (Willd.) R. Br. from different localities].

A comparative cytogenetic study of two introduced forms of Makleaya cordata (Willd.) R. Br. = syn. Bocconia cordata Willd. grown in different ecological and geographical regions (Moscow and Donetsk areas) was carried out. In the study, a complex of methods utilizing various chromosomal markers, i.e., C- and DAPI-banding technique, fluorescence in situ hybridization (FISH) with probes of26S and 5S rDNA, as well as estimation of the total area of C-positive regions (C-HCH) in prophase nucleoli and meiosis analysis, was used. In the karyotypes (2n = 20), each chromosome was identified on the basis of C-banding and FISH patterns and the chromosome ideograms were built. Pericentrometric and telomeric C-positive bands in chromosomes of the Moscow form karyotype were found to be smaller and intercalary bands, larger than the corresponding bands in the M. cordata form grown in Donetsk. It was found that the content of C-HCH in prophase nucleoli in the form of M. cordata grown in Donetsk was higher than in the form grown in Moscow. In both forms sites of 26S rDNA and 5s rDNA were localized on satellite chromosome 1 and on chromosome 4 respectively but the signals were more intensive in the plant form grown in Donetsk. The results of this study enable selecting M. cordata forms for use in pharmacology and recommending them for cultivation in various ecological and geographical regions.

[Modification of collagen during action light].

In work chemical modification of collagen during action visible spectrum sunlight. These changes of collagen were found to indicate a photodegradation and photooxidation processes in collagen.

[The use of nanoparticles for the study of conformations submembrane hemoglobin].

The advantages and features of the integrated application of methods of atomic force microscopy, laser interference microscopy and Raman microscopy in the study of erythrocytes was discussed. For the successful application of Raman microscopy in surface enhanced Raman spectroscopy mode the silver colloids was using. The dependence of the enhancement of Raman signals from silver colloids size was demonstrated. The using of developed methods for clinic diagnostic was discussed.

[Composite phytoadaptogens in oncology and gerontology].

The article describes results of research devoted to Phytomix-40, a mixture of plant adaptogens. It focuses on immunobiological criteria for its formulation, chemical composition and manufacture procedures, biological standartization tests, in vitro and in vivo preclinical studies, clinical trials in patients with non-malignant tumours (benign prostatic hyperplasia), precancer (oral leukoplakia), advanced cancer (malignant gastric cancer), and age-related neurodegenerative disease (parkinsonism). Prospects for the development of other plant preparations for non-toxic prevention and treatment of cancer and prolongation of life span of the affected subjects are discussed.

[Liquid-crystalline dispersions formed by complexes of linear double-stranded DNA molecules with dendrimers].

The formation of liquid-crystalline dispersions as a result of interaction of linear, double-stranded DNA molecules with poly(propyleneimine) dendrimers in water-salt solutions was studied. It was shown, that this process does not depend on the ionic strength of solution and molecular structure of dendrimer. By means of the atomic force microscopy, it was established, that in the case of the dendrimer molecules of the 4th generation (G4), the mean size of particles of (DNA-dendrimer G4) liquid-crystalline dispersion is equal to 300-400 nm. The "boundary" conditions (ionic strength of solutionand molecular mass of dendrimer) of formation of optically active (cholesteric) and optically inactive of the (DNA-dendrimer) dispersions were determined using circular dichroism spectroscopy. The interaction of dendrimers of 1st, 2nd, 3rd and 5th generations with DNA molecules results in the obtaining of the optically inactive dispersions. Dendrimer molecules of 4th generation induce the formation of two types dispersions: in solutions of high ionic strength (micro > 0.4) they induce the formation of cholesteric liquid-crystalline dispersions, and in solutions of low or intermediate ionic strength (micro < 0.4) they can form the optically inactive one. The "molecular crowding" affects both the efficiency of binding of dendrimer molecules of 4th generation to DNA, and the mode of spatial packing of (DNA-dendrimer G4) complexes in particles of liquid-crystalline dispersion. The possible reasons capable of explaining the structural polymorphism of (DNA-dendrimer) liquid-crystalline dispersions are discussed.

[Biochemical and photometric studies of modification of collagen structure under UV radiation].

The influence of UV irradiation (270-380 nm) on the biochemical, fluorescence and colorimetric properties of collagen was studied. The long-term UV irradiation (120 h) was accompanied by the increase of structural stability of collagen to specific and nonspecific proteolytic enzymes, by formation of new additional fluorophore-containing molecules, by the increase in quantity of carbonyl groups in the collagen and by strong changing of the distribution pattern of alkaline hydrolysis products during gel-chromatography. The coordinates of colour of the collagen films are also changed. These changes of collagen suggest that UV irradiation induces photomodification and photooxidation processes in collagen.

Comparative study of the state of water in various human tissues.

We studied the dependence of water exchange in various human tissues (skin, Achilles tendon, various types of hyaline cartilage, muscle, and muscle fascia) on the content of collagen and proteoglycans. Carbohydrates of the connective tissue do not play the major role in tissue water exchange. Collagen fibers and proteoglycan aggregates probably retain the main part of bound and free water, respectively.

Tissue specific expression of hepatitis B virus surface antigen in transgenic plant cells and tissue culture.

The tobacco plants (Nicotiana tabacum L.) carrying the HBsAg gene controlled by (Aocs)3AmasPmas, the hybrid promoter that includes regulatory elements of the agrobacterial octopine and mannopine synthase genes, as well as plants controlled by the same promoter and adh1, maize alcohol dehydrogenase gene intron were obtained. The presence of the adh1 gene intron did not significantly change the level of expression of the HBsAg gene in plants. The analysis of expression of hepatitis B virus surface antigen (HBs-antigen) in transformed plants expressing the HBsAg under the control of different promoters was made. The level of HBs-antigen in plants carrying the HBsAg gene controlled by (Aocs)3AmasPmas, the hybrid agrobacterium-derived promoter, was the highest in roots and made up to 0.01% of total amount of soluble protein. The level of HBs-antigen in plants carrying the HBsAg gene controlled by the dual 35S RNA cauliflower mosaic virus promoter was the same in all organs of the plant and made up to 0.06% of the total amount of soluble protein. Hairy root and callus cultures of plants carrying the HBsAg gene and expressing the HBs-antigen were obtained.

[Investigation of ribosomes of E. coli and T. maritima by atomic force microscopy].

Subunits 70S, 50S, and 30S of ribosomes of E. coli and T. maritima have been studied by atomic force microscopy. A considerable heterogeneity of structures was visualized when 70S and 30S subunits were sorbed on mica. The linear size and the height of molecules were estimated. It was found that the heights of ribosomes of E. coli and T. maritima substantially differ. The average height of 70S ribosomes of E. coli was 9.4 + 0.01 nm and that of T. maritima was 10.35 +/- 0.02 nm. The differences in the dimensions were probably determined by special organization of the mobile ribosomal element the L7/L12-stalk.

[A quantitative analysis of complex formation between IgM and immobilized ligand using atomic force microscopy]. / Kolichestvennyi analiz obrazovaniia kompleksov IgM s immobilizovannym ligandom s pomoshch'iu atomno-silovoi mikroskopii.

Specific interaction between human IgM and polyclonal antibodies immobilized on support was studied by atomic force microscopy. Human IgMs are responsible for a number of side effects arising during the xenotransplantation of mammalian organs to man. On the basis of atomic force microscopy, a quantitative analysis of complexes with IgM was performed. The data of the analysis agree well with the results of enzyme immunoassay. It was shown that the method of detection of immune complexes based on atomic force microscopy is able to detect specific antibodies/antigens in serum.

[Detection of immune complexes using atomic force microscopy]. / Vyiavlenie immunnykh kompleksov s pomoshchiu atomno-silovoi mikroskopii.

Complex formation between immunoglobulins and ligands immobilized on mica was studied by atomic force microscopy in two different systems. In the first system, 60-kDa ligands possessing only one site for antibody recognition were used. In the other system, a more complex interaction of human immunoglobulin with immobilized polyclonal antibodies was studied. In both systems, specific complexes with proper ligand appeared, and unspecific interaction was not detected. The method of revealing immunocomplexes by image atomic force microscopy can be used in the development of modern diagnostic systems.

Expression and characterization of hepatitis B surface antigen in transgenic potato plants.

Transgenic potato plants expressing the gene of hepatitis B surface antigen (HBsAg) under the control of the double promoter of 35S RNA of cauliflower mosaic virus (CaMV 35SS) and the promoter of patatin gene of potato tubers have been obtained. Biochemical analysis of the plants was performed. The amount of HBsAg in leaves, microtubers, and tubers of transgenic potatoes growing in vitro and in vivo was 0.005-0.035% of the total soluble protein. HBsAg content reached 1 microg/g in potato tubers and was maximal in plants expressing the HBsAg gene under the control of CaMV 35SS promoter. In transgenic plants expressing HBsAg gene under the control of tuber-specific patatin promoter, HBsAg was found only in microtubers and tubers and was absent in leaves. Western blot analysis of HBsAg eluted from immunoaffinity protein A-Sepharose matrix has been performed. The molecular weight of HBsAg peptide was approximately 24 kD, which is in agreement with the size of the major protein of the envelope of hepatitis B virus. Using gel filtration, it was determined that the product of HBsAg gene expression in potato plants is converted into high-molecular-weight multimeric particles. Therefore, as well as in recombinant HBsAg-yeast cells, assembling of HBsAg monomers into immunogenic aggregates takes place in HBsAg-transgenic potato, which can be used as a source of recombinant vaccine against hepatitis B virus.

[Phagocytosis of polymer microspheres immobilized with bioligands by peripheral blood leukocytes in patients with acute pancreatitis]. / Fagotsitirovanie polimernykh mikrosfer, immobilizovannykh bioligandami, leikotsitami perifericheskoi krovi u bol'nykh ostrym pankreatitom.

Phagocytosis of polymer microspheres both modified and unmodified by bioligands (gelatin, fibronectin and A protein) by leukocytic blood cells of patients with acute pancreatitis. The presence of bioligands in the surface of polymer microspheres and their activity were monitored by the reaction of latex agglutination with an appropriate ligand being involved. The study resulted in designing a method of phagocytosis implementation that can be used to evaluate the content of certain receptors in cellular membranes and, subsequently, to define the changed receptor cell apparatus caused by a disease. The nature of polymer microspheric bioligands immobilized to the surface was shown to influence significantly the activity of leukocytic blood cells.

[Structural features of proteins by intermittent-contact atomic force microscopy]. / Issledovanie strukturnykh osobennostei belkov preryvisto-kontaktnym metodom atomno-silovoi mikroskopii.

Optimal conditions of protein scanning by atomic force microscopy were developed. Proteins of different molecular masses (950-11.5 kDa) and different three-dimensional organization were used to investigate the structural features of proteins. The most distinct images of proteins were obtained using a tip with the free amplitude in the range of 5-15 nm and with the set-point amplitude in the regime of repulsion from the sample. The method allowed one to clearly recognize the structural details of large molecules such as immunoglobulins IgM and IgG1 and Ricinus agglutinin. The revealing of the structural properties of proteins with molecular masses of 60 kDa and less was limited by the sharpness of probe tips used in the present study. It was shown that, by a quantitative analysis of the geometric parameters of molecules, it is possible to distinguish IgG1, Ricinus agglutinin, and ricin.

Characteristics of collagenolytic enzymes secreted by deuteromycete fungi Aspergillus flavus.

Collagen induced the synthesis of collagenolytic enzymes in the culture of Aspergillus flavus. Enzyme activity of the culture increased during storage and passage on a medium with this inductor. We developed a method for isolation and purification of collagenolytic enzymes and obtained two electrophoretically homogenous enzyme preparations belonging to neutral thermolabile collagenolytic metalloproteinases.

Water status and LPO in rat tissues during massive blood loss and irradiation with He-Ne laser.

We revealed a relationship between water balance and LPO in the myocardium, liver, and blood plasma during massive blood loss and irradiation with He-Ne laser. Low-intensity laser irradiation of the plasma inhibits LPO and normalizes water balance in rat tissues during massive blood loss.

Phospholipid composition of granulation and fibrous tissues in rats after local application of melatonin.

We studied the effect of melatonin applied locally in doses of 1.5 and 15 mg/ml on the phospholipid composition of granulation and fibrous tissues in rats at various stages of regeneration. Melatonin (especially in a dose of 1.5 mg/ml) increased the total phospholipid content on day 8 of regeneration, which was primarily related to accumulation of ethanolamine- and serine-containing fractions. Melatonin in a dose of 15 mg/ml increased the concentration of choline-containing fractions on day 5 of regeneration.

Comparison of cytotoxicity of aminoglycoside antibiotics using a panel cellular biotest system.

The cytotoxicity of four aminoglycoside antibiotics was studied by estimation of the dose-effect relationship using a panel cellular biotest system including cell cultures for test objects. The cultures represented 4 differentiation types: normal human fibroblasts and myoblasts, human or Syrian hamster hepatoma cells, and mouse/mouse hybridoma cells. It was found that three widely used antibiotics gentamicin, kanamycin, and neomycin exhibit similar, but not identical cytotoxicity parameters and differ distinctly from geneticin. Hence, the proposed panel biotest system helps to quantitatively evaluate and differentiate the effects of bioactive substances with similar chemical structure.