[Identification of orthopoxvirus species using oligonucleotide micro-chips]. / Vidovaia identifikatsiia ortopoksvirusov na oligonukleotidnykh mikrochipakh.

A method for describing the Orthopoxviruses that are pathogenic both to man and animals is described in the article. The method is based on hybridization of a fluorescently labelled amplified DNA sample with oligonucleotides, which were immobilized in a microchip. Species-specific regions within the crmB gene encoding a viral analogue of the tumor necrosis factor receptor, i.e. an important gene determining the pathogenicity of the mentioned Orthopoxviruses type, were used as a target for identification. The identification procedure takes around 6 hours and does not demand any costly equipment (a portable fluorescent microscope can be used).

[Thermostability of alkaline phosphatase of Meithermus ruber bacteria: cloning of the gene, expression in Escherichia coli cells and biochemical characterization of the recombinant protein]. / Termostabil'naia shchelochnaia fosfataza bakterii Meiothermus ruber: klonirovanie gena, ékspressiia v kletkakh Escherichia coli i biokhimicheskaia kharakteristika rekombinantnogo belka.

The Meiothermus ruber alkaline phosphatase gene was cloned, expressed in Escherichia coli cells, and sequenced. The enzyme precursor, including the putative signal peptide, was shown to consist of 503 residues (deduced molecular mass 54,229 Da). The recombinant enzyme showed the maximal activity at 60-65 degrees C and pH 11.0 and had K(m) = 0.055 mM as estimated with p-nitrophenyl phosphate (pNPP). The enzyme proved to be moderately thermostable, retaining 50% activity after 6 h incubation at 60 degrees C and being completely inactivated in 2 h at 80 degrees C. In substrate specificity assays, the highest enzymic activity was observed with pNPP and dATP. Vanadate, inorganic phosphate, and SDS inhibited M. ruber alkaline phosphatase, while thiol-reducing agents had virtually no effect. The enzymic activity strongly depended on exogenous Mg2+ and declined in the presence of EDTA.

[Clinical value of microchip technology in determination of drug resistance of Mycobacterium tuberculosis]. / Klinicheskoe znachenie mikrochipovoi tekhnologii opredeleniia lekarstvennoi ustoichivosti mikobakterii tuberkuleza.

The patients with multiresistant tuberculosis were divided into 2 groups: the sensitivity of Mycobacteria tuberculosis to antituberculous drugs was evaluated in Group 1 by the methods of absolute concentrations and in Group 2 by biological microchips determining mutations in the rpo3 gene responsible for rifampicin resistance. The results of the drug sensitivity test were obtained after 3 months of treatment in Group 1 and several days prior treatment in Group 2. By taking into account the test results, reserve drugs was used in Group 2 patients. Subsequently, the results of the drug sensitivity tests carried out by the bacteriological method in Group 2 patients showed that isoniazid resistance was simultaneously noted if there were mutations in the rpo-B gene. Timely treatment with reserve drugs exhibited higher efficiency of treatment with its shorter duration in Group 2 than in Group 1.

[Use of bacterial luciferase for determining monoamine oxidase activity]. / Ispol'zovanie bakterial'noi liutsiferazy dlia opredeleniia monoaminooksidaznoi aktivnosti.

A bioluminescence assay is proposed for measuring monoamine oxidase activity in different biological specimens (platelets, mitochondria). The assay is based on the bioluminescent reaction catalysed by bacterial luciferase and coupled to monoamine oxidase. Two modifications of the bioluminescence assay were used. In the first case, the bioluminescent system was added to monoamine oxidase preincubated with the substrates, while in the second case, all the components of the coupled enzymatic systems were directly mixed in a cell. The proposed bioluminescence assay is simple, highly sensitive and rapid, and could be especially useful for biomedical examinations.