PLAGUE INFECTION SIMULATING IN CASE OF INOCULATION WITH AVIRULENT YERSINIA PESTIS STRAINS

Biological method of investigation is specified for the laboratory diagnostics of plague. Mastering of this method by the trainees within the frames of further vocational education is associated with the use of avirulent Yersinia pestis strains and vaccine Y. pestis strain EV line, which while providing safety does not allow for typical pathomorphological pattern on biomodels, as well as for isolation of microorganisms from internal organs. Objective of the study is to select avirulent Yersinia pestis strains and to conduct comparative analysis of the simulation techniques for plague on biomodels. Materials and methods. Utilized were Y. pestis strains. Virulence was evaluated both, in vitro (polymerase chain reaction) and in vivo (LD50 for white mice). Results and conclusions. Set forward have been avirulent Y. pestis strains, prospective in terms of mastering biological method of laboratory diagnostics of plague, and means of their application for simulating plague in biomodels. The designed approach allows for exercising biological methods of plague investigation to the fullest extent, enhancing biological safety of practical studies and reducing the time line for isolation and accumulation of pure bacterial culture

Антимикробная активность препаратов прополиса при проведении экспертизы качества лекарственных средств

The present article describes the analysis of the results of pharmaceutical expert evaluation of propolis preparations, performed in the laboratory of microbiology of the FSBI «SCEMP» of the Ministry of Health of the Russian Federation for the period of 2006-2014 in terms of «Antimicrobial activity». During the mention period 43 batches of medicinal products were analyzed, a significant part o which were solid domestic drugs (93%). Experimentally determined the effect of individual factors, such as the type of microorganism, the concentration and choice of nutrient medium, on the result of the analysis for this terms.Представлен анализ результатов фармацевтической экспертизы качества препаратов прополиса, проведенной в лаборатории микробиологии ФГБУ «НЦЭСМП» Минздрава России в период 2006-2014 гг. по показателю «Антимикробная активность». За указанный период проанализировано 43 серии лекарственных средств, значительную часть из которых составили препараты отечественного производства (93%). Экспериментально определено влияние отдельных факторов, таких как вид микроорганизма, его концентрация и выбор питательной среды, на результат анализа по данному показателю

Intraspecific Differentiation of <i>Francisella tularensis</i> Strains Using Multilocus Real-Time Polymerase Chain Reaction

The aim of the study was to develop a method for intraspecific differentiation of the tularemia microbe: subspecies tularensis (subpopulations AI and AII), holarctica (biovars japonica, EryS/R), mediasiatica, and novicida using multilocus real-time PCR. Materials and methods. We used 48 strains of F. tularensis of various subspecies, biovars, and subpopulations. Intraspecific appurtenance of the strains was carried out on the basis of the analysis of the RD-1 region variability applying PCR, the sdhA gene by Sanger fragment sequencing and by the disk diffusion method using disks with erythromycin. The selection of primers and probes was performed using the software available at www.genscript.com and GeneRunner 6.5.52. Sequence homology was assessed using the BLAST algorithm and the GenBank NCBI database. Results and discussion. New data on the structure and occurrence of the differentiation regions RD-8, RD-12, RD-28 of FTT1122c gene and its homologous sequences in strains of tularemia microbe of various subspecies have been obtained. Novel RDhm 346 bp in size, characteristic of strains of the subsp. mediasiatica, holarctica, which is deleted in subsp. tularensis and absent in subsp. novicida has been detected. Based on the detection of the FTT1670, FTT1122с, FTT1067, FTW_2084 loci, a multilocus real-time PCR has been developed – “F. tularensis 4c”, providing for identification of all subspecies of the tularemia microbe, separately for the biovar japonica of the Holarctic subspecies and subpopulations AI, AII of the subspecies tularensis. The PCR specificity was confirmed in the study of strains of tularemia microbe from the fund of the “State Collection of Pathogenic Bacteria” at the premises of the Russian Reserarch Anti-Plague Institute “Microbe”. The results obtained expand the concept of intraspecific genetic heterogeneity of tularemia microbe and possibilities of identifying the causative agent of tularemia using molecular-genetic methods. They are important for understanding the processes of adaptation of the pathogen to circulation in the host organism and environmental objects, the course of evolution and formation of new species of Francisella

Development of a Method for Determination of brucella suis Biovars Using Multilocus Real-time PCR

The aim of the study was to develop a methodological approach to determination of Brucella suis biovars through multilocus PCR with real-time registration of results.Materials and methods. We used 16 strains of B. suis of various biovars, B. neotomae and B. canis – 2 strains of each. Determination of the taxonomic affiliation of Brucella strains was carried out according to the Bruce-ladder, Suis-ladder, BRU-DIF protocols. The selection of primers and probes was performed using the software on the website www.genscript.com and the GeneRanner 6.5.52 program. Fragment sequencing according to Sanger was performed on a 3500 XL genetic analyzer in accordance with the manufacturer’s recommendations. Nucleotide sequence homology was assessed using the BLAST algorithm and the GenBank NCBI database.Results and discussion. An analysis of the structural organization of IncP and GI-3 genomic islands has been carried out in B. suis strains of various biovars. It has been established that in strains of B. suis II, IV biovars and B. canis, the terminal part of the BRA0368 gene, comprising 21 nucleotides (repeated in the BRA0367 gene) and the “TAA” stop codon, as well as almost the entire sequence of the BRA0367 gene were lost, owing to homologous recombination in the IncP genome island. A 21-nucleotide direct repeat and the “TGA” stop codon of the BRA0367 gene replaced the analogous region of the BRA0368 gene which resulted in the deletion the size of 185 bp. No differences have been noted in the structure of GI-3 in biovars. The evidence obtained made it possible to develop the approach (SuisDIF) for differentiating B. suis biovars, based on the amplification of genes located in the IncP and GI-3 genomic islands using real-time PCR. Its specificity was confirmed in the study of B. suis strains from the fund of the State Collection of Pathogenic Bacteria of the Russian Research Anti-Plague Institute “Microbe”. The conducted studies expand and supplement the data on the genetic heterogeneity of Brucella species and biovars. The proposed method for differentiating biovars of B. suis using multilocus PCR with real-time registration of results enhances the capacities for Brucella identification using molecular-genetic methods

ЛЕКАРСТВЕННЫЕ СРЕДСТВА, СОДЕРЖАЩИЕ ЗМЕИНЫЙ ЯД: ИСТОРИЯ РАЗВИТИЯ, НОМЕНКЛАТУРА, ОЦЕНКА ПОДЛИННОСТИ

The article presents general data on the chemical composition and properties of snake venoms, as well as the history of their use in medical practice. It considers the differences characteristic of the venom of snakes belonging to different systematic groups. It demonstrates that venoms remain a valuable source of new medicines. The article compares existing methods of venom analysis (immunological, chromatographic, electrophoretic, mass spectrometric). Particular attention is paid to specific metho-dological aspects of identification testing depending on the object and purpose of the analysis. The article also compares different modifications of the precipitation reaction. It highlights the advantages and disadvantages of existing methods. The affordability and simplicity of immunological methods accounts for their wide use in the analysis of poisons in biological media which helps to determine the cause of poisoning. While greater informativeness of instrumental methods makes them a perfect toll for detailed examination of poison composition during poisons research and comparison. The authors of the article performed  a retrospective analysis of data obtained in identification testing of snake venoms during the period from 2006 to 2017. It has been established that manufacturer specifications for individual medicinal products in some cases do not contain instructions on the use of a specific type of precipitation reaction or the description of test procedures. It was concluded that some modifications should be made to the existing method, the choice of conditions for sample preparation should be considered, or alternative methods for identification of snake venoms in medicinal products should be developed.Представлены общие данные о химическом составе, свойствах ядов змей и истории их применения в медицинской практике. Рассмотрены отличия, характеризующие яд змей той или иной систематической группы. Отмечено, что яды продолжают оставаться ценным источником новых лекарств. Проведено сравнение существующих методов анализа ядов (иммунологического, хроматографического, электрофоретического, масс-спектрометрического). Особое внимание уделено методическим особенностям определения их подлинности в зависимости от объекта и цели анализа. Выполнено сравнение различных модификаций реакции преципитации между собой. Выявлены преимущества и недостатки существующих методов. Показано, что иммунологические методы ввиду своей доступности и простоты исполнения чаще всего применяются при анализе ядов в биологических средах для диагностики источника отравления. А инструментальные методы благодаря большей информативности незаменимы для детальных исследований состава ядов при их изучении и сравнении. Проведен ретроспективный анализ данных, полученных при испытании подлинности змеиных ядов за период с 2006 по 2017 г. Установлено, что нормативная документация на отдельные лекарственные средства в ряде случаев не содержит указаний на использование конкретного вида реакции преципитации или описание методик. Сделан вывод о целесообразности модификации существующего метода, выбора условий пробоподготовки или разработки альтернативных методов для определения подлинности змеиных ядов, входящих в состав лекарственных средств

Ultrafast laser micro-nano structuring of transparent materials with high aspect ratio

Ultrafast lasers are ideal tools to process transparent materials because they spatially confine the deposition of laser energy within the material's bulk via nonlinear photoionization processes. Nonlinear propagation and filamentation were initially regarded as deleterious effects. But in the last decade, they turned out to be benefits to control energy deposition over long distances. These effects create very high aspect ratio structures which have found a number of important applications, particularly for glass separation with non-ablative techniques. This chapter reviews the developments of in-volume ultrafast laser processing of transparent materials. We discuss the basic physics of the processes, characterization means, filamentation of Gaussian and Bessel beams and provide an overview of present applications

Modularity in Protein Complex and Drug Interactions Reveals New Polypharmacological Properties

Recent studies have highlighted the importance of interconnectivity in a large range of molecular and human disease-related systems. Network medicine has emerged as a new paradigm to deal with complex diseases. Connections between protein complexes and key diseases have been suggested for decades. However, it was not until recently that protein complexes were identified and classified in sufficient amounts to carry out a large-scale analysis of the human protein complex system. We here present the first systematic and comprehensive set of relationships between protein complexes and associated drugs and analyzed their topological features. The network structure is characterized by a high modularity, both in the bipartite graph and in its projections, indicating that its topology is highly distinct from a random network and that it contains a rich and heterogeneous internal modular structure. To unravel the relationships between modules of protein complexes, drugs and diseases, we investigated in depth the origins of this modular structure in examples of particular diseases. This analysis unveils new associations between diseases and protein complexes and highlights the potential role of polypharmacological drugs, which target multiple cellular functions to combat complex diseases driven by gain-of-function mutations