ПРИМЕНЕНИЕ MALDI МАСС-СПЕКТРОМЕТРИИ В ДИАГНОСТИКЕ ОСОБО ОПАСНЫХ ИНФЕКЦИОННЫХ БОЛЕЗНЕЙ: СОВРЕМЕННОЕ СОСТОЯНИЕ И ПЕРСПЕКТИВЫ ФКУЗ «Российский научно-исследовательский противочумный институт «Микроб», Саратов, Российская ФедерацияМасс-спектрометрия является современным физико-химическим методом анализа, позволяющим проводить качественный и количественный анализ состава вещества, основанный на предварительной ионизации входящих в его состав атомов или молекул. Одним из новых методов ионизации, благодаря которому масс-спектро метрическое исследование макромолекул получило широкое распространение, является разработанная матричноактивированная лазерная десорбция/ионизация (MALDI), представляющая собой импульсное лазерное облучение исследуемого вещества, смешанного с матрицей. В обзоре представлены современные данные о применении метода MALDI масс-спектрометрии для проведения родо-и видоспецифической идентификации микроорганизмов в практике диагностических лабораторий. Рассмотрены преимущества MALDI-TOF идентификации по сравнению с бактериологическими, иммунологическими и молекулярно-генетическими методами исследования. Обозначено место масс-спектрометрии в системе лабораторной диагностики инфекционных болезней, в том числе особо опасных на территории Российской Федерации.Ключевые слова: MALDI масс-спектрометрия, MALDI-TOF, идентификация, патогенные биологические агенты. A.N.Spitsyn, D.V.Utkin, V.E.Kuklev, S.A.Mass spectrometry is a modern physical-chemical analytical method that provides for qualitative and quantitative assessment of the substance composition. It is based on pre-ionization of the atoms and molecules included into it. One of the advanced methods of ionization, due to which mass-spectrometry investigation of macromolecules has become a frequent practice, is matrix-assisted laser desorption/ionization (MALDI). The essence of it is the pulsed laser irradiation of the matter under study, mixed with the matrix. The review discusses current data on MALDI mass-spectrometry application for the performance of species-specific and genus-specific identification of microorganisms at the premises of diagnostic laboratories. Considered are the basic advantages of MALDI-TOF identification as compared to bacteriologic, immunologic, and molecular-genetic methods of assessment. Allocated is the mass-spectrometry position in the system of laboratory diagnostics of infectious diseases, including particularly dangerous ones, in the territory of the Russian Federation.
Acute poisoning with alcohols and cholinotropic preparations carboxyphosphamide and atropine (0.8 LD(50)) was modeled on male outbred mice weighing 18-24 g. The decrease in activity of natural killer cells was most pronounced after injection of atropine, but insignificant after treatment with ethanol. The inhibitory effect of ethylene glycol, methanol, and methanol on functional activity of natural killer cells in vitro directly depended on their concentration. The effects of alcohols in equimolar concentrations of 10, 100, and 500 mM were similar. Therefore, immunotoxicity of alcohols was associated with the action of their metabolites. The ability of products formed after biotransformation of ethylene glycol, methanol, and ethanol in equimolar concentrations to cause damage to natural killer cells decreased in the following order: glyoxylic acid>formic acid>acetaldehyde>glycolaldehyde>glycolic acid. T-Activin injected subcutaneously in doses of 2.5 and 5.0 microg/kg for 3 days normalized activity of natural killer cells suppressed after acute poisoning with alcohols and cholinotropic preparations.
Experiments on Wistar rats showed that acute poisoning with organophosphorus compound dimethyl dichlorovinyl phosphate (0.5 LD(50)) was accompanied by two opposite effects: inhibition of acetylcholinesterase in T lymphocytes leading to suppression of thymus-dependent antibody formation (predominant effect) and acetylcholine-induced stimulation of antibody production. Acetylcholine activated acetylcholinesterase in intact T cells.
Acute poisoning with tetrachloromethane in a dose of 0.75 LD(50) suppressed humoral and cell immune reactions in Wistar rats. Immunotoxicity of tetrachloromethane is realized via initiation of lipid peroxidation and inhibition of acetylcholine esterase in T lymphocytes and alpha-naphthyl AS-acetate esterase and alpha-naphthyl butyrate esterase in splenocytes.
Experiments on Wistar rats showed that subacute poisoning with anticholinesterase toxicants zarin and agent VX (daily subcutaneous injections in 1/7 LD50 for 6 days) led to suppression of cellular and humoral immune reactions and to a decrease in blood concentrations of cytokines (IL-2, IL-4, IFN-gamma) with a reduction of the IFN-gamma/IL-4 and IL-2/IL-4 ratios, which attests to more pronounced decrease in Th1 lymphocyte function in comparison with Th2 cells.
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