RESUMEN
G-quadruplexes (G4), non-canonical secondary DNA structures, are intensively investigated for a long time. In eukaryotic organisms they play an important role in the regulation of gene expression and DNA repair. G4 have also been found in the genomes of numerous bacteria and archaea, but their functional role has not yet been fully explored. Nevertheless, their participation in the formation of antigenic variability, pathogenicity, antibiotic resistance and survival in extreme conditions has been established. Currently, many tools have been developed to detect potential G4 sequences and confirm their formation ability. Since the controlled formation and resolution of the quadruplex are significant means for the regulation of genes critical for survival, a promising direction is the search for ligands - compounds that can have a stabilizing effect on the quadruplex structure and thereby alter gene expression. Currently, a number of ligands are already known, their use stops the growth of pathogenic microorganisms. G4 ligands are of interest as potential antibiotics, which are extremely relevant due to the wide spread of drug resistant pathogens.
Asunto(s)
G-Cuádruplex , Bacterias/genética , ADN/química , LigandosRESUMEN
Comparative proteomic profiling of M. tuberculosis H37Rv strains cultured on two different nutrient media, Levenstein-Jensen and Middlebrook 7H11, was performed using a label-free LC-MS/MS approach. It was shown that results obtained from two media possessed high convergence. The only difference was observed in the representation of fumarate reductase FrdB, its abundance was higher in the mycobacterial cells cultured on Levenstein-Jensen medium. The correlation analysis of biological repeats revealed the high convergence of the results obtained from Middlebrook 7H11 medium. Thus, we can conclude that the use of the Middlebrook 7H11 medium is most appropriate in the scientific laboratory.
Asunto(s)
Mycobacterium tuberculosis/metabolismo , Proteoma/metabolismo , Proteínas Bacterianas/metabolismo , Cromatografía Liquida , Medios de Cultivo , Succinato Deshidrogenasa/metabolismo , Espectrometría de Masas en TándemRESUMEN
The physiology of Mycobacterium tuberculosis, the causative agent of tuberculosis, is being studied with intensity. However, despite the genomic and transcriptomic data available today, the pathogenic potential of these bacteria remains poorly understood. Therefore, proteomic approaches seem relevant in studying mycobacteria. This review covers the main stages in the proteomic analysis methods used to study mycobacteria. The main achievements in the area of M. tuberculosis proteomics are described in general. Special attention is paid to the proteomic features of the Beijing family, which is widespread in Russia. Considering that the proteome is a set of all the proteins in the cell, post-translational modifications of mycobacterium proteins are also described.
RESUMEN
INTRODUCTION: Pancreatic necrosis - acute surgical disease accompanying high rates of mortality and complications. MATERIAL AND METHODS: The analysis of the treatment of 256 patients with pancreatic necrosis moderate and severe degrees. In the control group was conducted basic therapy, the main (n=158), which further were administered high doses of octreotide and endoscopic stenting of the main pancreatic duct to decrease the intraductal pressure, prevention and treatment of internal pancreatic fistula. RESULTS AND DISCUSSION: In the main group in contrast to the control is marked by a more rapid decline in the level of amylase, lipase and leukocyte index of intoxication. The infection occurred in the basic - 11%, control 20% died in the study group - 3%, control - 9%.
Asunto(s)
Endoscopía Gastrointestinal , Octreótido/administración & dosificación , Páncreas , Fístula Pancreática , Pancreatitis Aguda Necrotizante , Complicaciones Posoperatorias/prevención & control , Terapia Combinada/métodos , Descompresión Quirúrgica/instrumentación , Descompresión Quirúrgica/métodos , Endoscopía Gastrointestinal/efectos adversos , Endoscopía Gastrointestinal/instrumentación , Endoscopía Gastrointestinal/métodos , Femenino , Fármacos Gastrointestinales/administración & dosificación , Humanos , Masculino , Persona de Mediana Edad , Evaluación de Procesos y Resultados en Atención de Salud , Páncreas/patología , Páncreas/cirugía , Fístula Pancreática/etiología , Fístula Pancreática/prevención & control , Pancreatitis Aguda Necrotizante/diagnóstico , Pancreatitis Aguda Necrotizante/mortalidad , Pancreatitis Aguda Necrotizante/cirugía , Federación de Rusia/epidemiología , Índice de Severidad de la Enfermedad , StentsRESUMEN
Steadily growing resistance of the tuberculosis causative agent towards a broad spectrum of anti-tuberculosis drugs calls for rapid and reliable methods for identifying the genetic determinants responsible for this resistance. In this study, we present a biochip-based method for simultaneous identification of mutations within rpoB gene associated with rifampin resistance, mutations in katG, inhA, ahpC genes responsible for isoniazid resistance, mutations within the regions of gyrA and gyrB genes leading to fluoroquinolones resistance, and mutations in the rrs gene and the eis promoter region associated with the resistance to kanamycin, capreomycin and amikacin. The oligonucleotide microchip, as the core element of this assay, provides simultaneous identification of 99 mutations in the format "one sample--one PCR--one microchip", and it makes it possible to complete analysis of multi-drug-resistant and extensively drug-resistant tuberculosis within a single day. The tests on 63 Mycobacterium tuberculosis clinical isolates with different resistance profiles using the developed approach allows us to reveal the spectrum of drug-resistance associated mutations, and to estimate the significance of the inclusion of extra genetic loci in the determination of M. tuberculosis drug resistance.
Asunto(s)
Antituberculosos/farmacología , Farmacorresistencia Bacteriana Múltiple/genética , Regulación Bacteriana de la Expresión Génica , Ensayos Analíticos de Alto Rendimiento/instrumentación , Mycobacterium tuberculosis/efectos de los fármacos , Mycobacterium tuberculosis/genética , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Técnicas de Tipificación Bacteriana , Catalasa/genética , Catalasa/metabolismo , Girasa de ADN/genética , Girasa de ADN/metabolismo , ARN Polimerasas Dirigidas por ADN , Tuberculosis Extensivamente Resistente a Drogas/microbiología , Fluoroquinolonas/farmacología , Humanos , Isoniazida/farmacología , Kanamicina/farmacología , Procedimientos Analíticos en Microchip , Mutación , Mycobacterium tuberculosis/crecimiento & desarrollo , Mycobacterium tuberculosis/aislamiento & purificación , Oxidorreductasas/genética , Oxidorreductasas/metabolismo , Peroxirredoxinas/genética , Peroxirredoxinas/metabolismo , Rifampin/farmacologíaRESUMEN
A minisequencing method based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF MS) was developed for rapid identification of single nucleotide polymorphisms at bla(TEM) gene codons 104, 164 and 238 associated with extended-spectrum activity on TEM-type beta-lactamases. The method was validated by testing the Escherichia coli and Klebsiella pneumoniae strains possessing the known bla(TEM) gene sequences.