Role of N-acetyltransferase 2 gene polymorphism in the human pathology

Nowadays multiple heterogeneous chemicals affect the human body. They include drugs, household chemicals, dyes, food supplements and others. The human organism can modify, inactivate, and eliminate the chemicals by biotransformation enzymes. But it is well known that biotransformation can lead to toxification phenomenon. Individuals differ from each other by the rate of chemical modification that promotes accumulation of toxins and carcinogens in some patients. An N-acetyltransferase 2 enzyme participates in the aromatic amines second phase metabolism. This work reviews the acetyltransferase gene polymorphism possible role in diseases development including drug-induced organs damage.Gene of acetyltransferase has polymorphisms associated with two haplotypes of  fast and slow substrate acetylation. Gene alleles combine in three genotypes: fast, intermediate, and slow acetylators. Acetylation rate plays a significant role in side effects development during tuberculosis treatment and cancer pathogenesis. Recently, new data described the role of enzyme in development of non-infectious diseases in the human. Scientists consider that slow acetylation genotype in combination with high xenobiotic load result in accumulation of toxic substances able to damage cells.Therefore, acetyltransferase genotyping helps to reveal risk groups of cancer and non-infectious disease development and to prescribe more effective and safe doses of drugs

COMPARATIVE ANALYSIS OF CRISPR-CAS SYSTEM STRUCTURES OF YERSINIA PSEUDOTUBERCULOSIS IP32953 AND IP31758

Background. Pseudotuberculosis is still relevant problem in medical science and public health of Russia and other countries. Typing of Y. рseudotuberculosis strains by their CRISPR systems is a perspective tool for monitoring of Yersinia populations as was shown in Y. pestis.Aims. Here we describe and compare CRISPR-Cas systems of Yersinia pseudotuberculosis strains IP32953 and IP31758 causing classic pseudotuberculosis and Far-East scarlet-like fever (FESLF) respectively.Materials and methods. Complete genomes of Y. pseudotuberculosis IP329353 and IP31758 (NC_006155 and NC_009708 respectively) were obtained from NCBI Nucleotide Database. Search; identification; and analysis of CRISPR systems were carried out by online-tools CRISPROne; CRISPRDetect; and CRISPRTarget.Results and discussion. Analyzed strains have CRISPR-Cas systems that include one set of cas-genes and arrays situated at the long distances from each other. We defined three CRISPR arrays in Y. pseudotuberculosis IP32953 by the combination of program methods. CRISPR-Cas system of this strain consist of array YP1 located near cas-genes; arrays YP2 and YP3. CRISPR-Cas system of Y. pseudotuberculosis IP31758 includes two arrays – YP1 and YP3. CRISPR systems do not share similar spacers. CRISPR systems of the analyzed strains differ in CRISPR loci and cas-protein structures that can be used as specific marks of analyzed strains.Conclusions. We suggest that acquisition of certain spacers may play a role in evolution and divergence of Y. pseudotuberculosis strains

Bioinformational analysis of Yersinia pseudotuberculosis IP32953 CRISPR/cas system

The results of this study include Yersinia pseudotuberculosis CRISPR/Cas system structure analysis. CRISPR/Cas system is a specific adaptive protection against heterogeneous genetic elements. The object of research was the complete genome of Y. pseudotuberculosis IP32953 (NC_006155). CRISPR/Cas system screening was performed by program modelling methods MacSyFinder ver. 1.0.2. CRISPR loci screening and analyzing were carried out by program package: CRISPR Recognition tool (CRT), CR1SP1: a CRISPR Interactive database, CRISPRFinder, and PilerCR. Spacer sequences were used in order to find protospacers in ACLAME, GenBank-Phage and RefSeq-Plasmid databases by BLASTn search algorithm. Protospacer sequences could be found in genomes of phages, plasmids and bacteria. In last case complete genomes of bacteria were analyzed by online-tool PHAST: PHAge Search Tool. Y. pseudotuberculosis IP329353 has CRISPR/Cas system that consists of one sequence of cas-genes and three loci. These loci are far away from each other. Locus YP1 is situated in close proximity to cas-genes. Protospacers were found in genomes of Y. pseudotuberculosis PB1/+, Y. intermedia Y228, Y. similis str. 228, Salmonella phage, Enterobacteria phage, Y. pseudotuberculosis 1P32953 plasmid pYV and plasmid of Y. pseudotuberculosis 1P31758. Thus, the combination of four program methods allows finding CRISPR/Cas system more precisely. Spacer sequences could be used for protospacer screening

BIOINFORMATION SEARCH AND ANALYSIS OF STRUCTURES OF CRISPR/CAS SYSTEMS IN PHAGE STAPHYLOCOCUS AUREUS GENOME AND ESTIMATION OF PROFILES OF PHAGE DETECTED THROUGH CRISPR-CASSETTE BACTERIA

The emergence of resistance among the most important bacterial pathogens is generally recognized as one of the major public health problems. The most important of these organisms are penicillin-resistant Streptococcus pneumoniae, vancomycin-resistant enterococci and methicillin- and vancomycin-resistant Staphylococcus aureus. These antibiotic resistance in common pathogens have made antimicrobial therapy of many infections. Scientists need to look for new ways of treating bacterial infections in the work, using the developed algorithm from the methods of search software in the genomic structure of Staphylococcus aureus subsp. aureus ST228, the CRISPR/Cas locus and the division structures of its CRISPR cassette. The results of the bacteriophage search through the decoded spacer sequences of CRISPR-cassettes of this strain were also obtained using the developed algorithm of the software methods of bioinformatics. It was determined that the CRISPR/Cas system of strain of ST228 of S. aureus was of type IIIA. It is shown that cas-genes are in the immediate vicinity of CRISPR cassettes. The spacer structures in the detected CRISPR cassette are the Staphylococcus, Mycobacterium, Streptococcus, Bacillus, Gordonia, Arthrobacter, Streptomyces. The implementation of the algorithm of program methods for locating CRISPR/Cas-loci can be applied to many other decoded bacterial genomes to return bacteriophage therapy

In silico сравнительный анализ crispr-систем штаммов Yersinia pseudotuberculosis, вызывающих различные клинические проявления псевдотуберкулёза

The aim of this research was to analyze and compare CRIPSR loci and cas-proteins of Yersinia pseudotuberculosis strains isolated in different territories from patients with various clinical manifestations of pseudotuberculosis.Materials and Methods. Complete genomes of Y. pseudotuberculosis IP329353 (NC_006155) and IP31758 (NC_009708) were obtained from NCBI Nucleotide Database. Strains were isolated from patients with gastroenteritis and systemic infection respectively. Search, identification, and analysis of CRISPR systems were carried out by onlinetools CRISPROne, CRISPRDetect, and CRISPRTarget.Results. Analyzed strains have CRISPR-Cas systems that include one set of cas-genes and arrays situated at the long distances from each other. We defined three CRISPR arrays in Y. pseudotuberculosis IP32953: array YP1 located near cas-genes, arrays YP2 and YP3. CRISPR-Cas system of Y. pseudotuberculosis IP31758 includes two arrays – YP1 and YP3. CRISPR systems do not share similar spacers.Conclusion. CRISPR systems of the analyzed strains differ in CRISPR loci and cas-protein structures that can be used as specific molecular marks of analyzed strains during the study of intra-species variability and evolution of Y. pseudotuberculosis.Цель: сравнить CRISPR-системы двух штаммов, выделенных на различных территориях от пациентов с разными клиническими проявлениями псевдотуберкулеза, и определить специфические различия в спейсерном составе и в структуре cas-белков.Материалы и методы: проанализированы полногеномные последовательности штаммов Y. pseudotuberculosis IP329353 (NC_006155) и IP31758 (NC_009708) различного географического происхождения, выделенные от больных с псевдотуберкулезом с симптомами гастроэнтерита и системными проявлениями инфекции соответственно. Поиск, идентификация и анализ CRISPR систем выполнены с использованием онлайн-приложений CRISPROne, CRISPRDetect и CRISPRTarget.Результаты: в геноме исследуемых штаммов обнаружены CRISPR-Cas системы, включающие один набор cas-генов и несколько CRISPR-локусов, значительно удаленных друг от друга. В геноме штамма Y. pseudotuberculosis IP329353 присутствует три локуса: YP1, находящийся в непосредственной близости от cas-генов, YP2 и YP3. CRISPR-Cas система Y. pseudotuberculosis IP31758 представлена только двумя кассетами: YP1 и YP3. CRISPR системы исследуемых штаммов не имеют одинаковых спейсеров.Заключение: CRISPR-Cas системы исследованных штаммов отличаются количеством CRISPR-локусов, их спейсерным составом и структурой cas-белков. Полученные результаты определяют перспективу использования CRISPR-локусов в качестве специфических молекулярных маркеров штаммов при изучении внутривидового разнообразия и эволюции Y. pseudotuberculosis

CRISPR-Cas Loci of Yersinia pseudotuberculosis Strains with Different Genetic Determinants

Relevance. Yersinia pseudotuberculosis is a causative agent of pseudotuberculosis, a disease with polymorphism of clinical manifestation that is determined by the presence of specific virulence determinants: plasmid pVM82, pathogenicity islands HPI and YAPI, and superantigen YPM. Occurrence of new determinants depends on horizontal transfer of mobile genetic elements, hence, systems regulating horizontal transfer participate in evolution of pathogenic species. CRISPR-Cas is and adaptive protection system of prokaryotes against mobile genetic elements. Aim. The study analyzed an interaction between CRISPR-loci of Y. pseudotuberculosis and virulence determinants. Results. 86% of strains includes three CRISPR-loci: YP1, YP2, and YP3. Length of locus YP3 mostly depends on presence of virulence determinants in strains of Y. pseudotuberculosis serotype O:1b. Strains with virulence genes are able to cause a severe form of pseudotuberculosis and have longer locus than strains without determinants. Conclusion. Therefore, CRIPSRCas system of Y. pseudotuberculosis may participate in formation of a certain strain genotype that defines clinical manifestation of pseudotuberculosis