RESUMEN

For evaluation of effects of release-active antibodies to CD4 on cultured lymphocytes from human peripheral blood, we measured intracellular content of lck-kinase cell-based ELISA. In cells treated with release-active antibodies to CD4, the content of intracellular lck-kinase significantly (p<0.01) decreased in comparison with the control (purified water processed in a similar way). Phytohemagglutinin had no effect on the concentration of lck-kinase in cells. The decrease in the content of CD4-associated lck protein suggests that the preparation enhanced intracellular coupling of lck-kinase with T-cell receptor and potentiated T-cell immune response.

Asunto(s)

Anticuerpos/farmacología , Antígenos CD4/genética , Linfocitos T CD4-Positivos/efectos de los fármacos , Proteína Tirosina Quinasa p56(lck) Específica de Linfocito/genética , Adulto , Antígenos CD4/inmunología , Linfocitos T CD4-Positivos/citología , Linfocitos T CD4-Positivos/inmunología , Ensayo de Inmunoadsorción Enzimática , Femenino , Regulación de la Expresión Génica , Humanos , Activación de Linfocitos/efectos de los fármacos , Proteína Tirosina Quinasa p56(lck) Específica de Linfocito/inmunología , Masculino , Fitohemaglutininas/farmacología , Cultivo Primario de Células

RESUMEN

Antiviral activity of Ergoferon was studied in vitro on an experimental model of rotavirus infection in MA-104 cell line. In infected cells treated with Ergoferon, rotavirus titer was shown to decrease by 83 and 90% in comparison with cells treated with solvent used for Ergoferon preparation (p<0.05) and distilled water (p<0.05), respectively. These findings demonstrate high anti-rotavirus activity of Ergoferon.

Asunto(s)

Anticuerpos/farmacología , Antivirales/farmacología , Rotavirus/efectos de los fármacos , Carga Viral/efectos de los fármacos , Animales , Antígenos CD4/inmunología , Línea Celular , Embrión de Mamíferos , Histamina/inmunología , Interacciones Huésped-Patógeno , Interferón gamma/antagonistas & inhibidores , Riñón/efectos de los fármacos , Riñón/patología , Riñón/virología , Macaca mulatta , Pruebas de Sensibilidad Microbiana , Rotavirus/fisiología

RESUMEN

Asthma is among the most common chronic disorders of airways, which affects both children and adults. Asthma being a common disease among different segments of population, it has a high mortality rate and, in the absence of appropriate care, affects the quality of life and leads to economics losses. In a view of continuing growth in the incidence of asthma, it is important to find relevant biological targets for developing new approaches to astma therapy. Recent advances in molecular immunology, genetics, and bioinformatics allowed genes involved in the pathogenesis of asthma to be identified, which provided prerequisites for the development of new types of drugs that can regulate the activity of pathogenically significant genes. To date, a number of technologies for sequence-specific gene regulation (ASO, ribozymes, DNAzymes, EGS, DNA-decoys, U 1-adapters) are available, but RNA interference is the most promising approach in both terms of efficacy and financial cost. This review focuses on the generalization and analysis of experimental data regarding the use of RNA interference technology for the treatment of astma.

Asunto(s)

Asma/terapia , Factor de Transcripción GATA3/antagonistas & inhibidores , Terapia Genética/métodos , Interleucina-13/antagonistas & inhibidores , Interleucina-4/antagonistas & inhibidores , Interferencia de ARN , Adulto , Aptámeros de Nucleótidos/genética , Aptámeros de Nucleótidos/metabolismo , Aptámeros de Nucleótidos/uso terapéutico , Asma/genética , Asma/inmunología , Asma/patología , Niño , Enfermedad Crónica , ADN Catalítico/genética , ADN Catalítico/metabolismo , ADN Catalítico/uso terapéutico , Factor de Transcripción GATA3/genética , Factor de Transcripción GATA3/inmunología , Expresión Génica , Predisposición Genética a la Enfermedad , Humanos , Interleucina-13/genética , Interleucina-13/inmunología , Interleucina-4/genética , Interleucina-4/inmunología , Oligonucleótidos Antisentido/genética , Oligonucleótidos Antisentido/metabolismo , Oligonucleótidos Antisentido/uso terapéutico , Calidad de Vida , ARN Catalítico/genética , ARN Catalítico/metabolismo , ARN Catalítico/uso terapéutico

RESUMEN

The nucleotide sequences of three DNA fragments (total size 30574 bp) of the plasmid p19 from the Bacillus subtilis 19 soil strain have been determined. Thirty open reading frames (ORFs) have been identified in these fragments. oriT of the plasmid has also been identified. As shown by the search for homologs of hypothetical protein products of these ORFs in databases, such homology exists for 18 ORFs. The protein products of nine ORFs can be assumed to have specific functions. Several ORFs were inactivated via insertional mutagenesis, and the conjugation capacity of the mutant plasmids was estimated. According to the data on homology of protein products and the results of ORF inactivation, regions of a total size of about 20 kb from the DNA fragments sequenced by us were inferred to belong to the tra region of p19. As follows from the analysis of the identified ORFs of the p19 tra region, it differs from the earlier described tra regions of other plasmids, irrespective of a certain similarity with the corresponding regions of plasmids of gram-positive bacteria from the genera Bacillus, Clostridium, and Listeria.

Asunto(s)

Bacillus subtilis/genética , Conjugación Genética , Plásmidos/genética , Microbiología del Suelo , Bacillus/genética , Secuencia de Bases , Clostridium/genética , Listeria/genética , Datos de Secuencia Molecular , Sistemas de Lectura Abierta/genética , Origen de Réplica , Análisis de Secuencia de ADN

RESUMEN

Interfering RNA (RNAi) is a powerful tool to silence gene expression on the level of mRNA. To knock-down gene expression by using RNAi two major methods of mRNA silencing exist. First method utilizes siRNA (small interfering RNA), a readily processed dsRNA, that enters RISC complex and destroy target mRNA after transfection into the cells. The second method based on the construction of plasmid DNA that expresses shRNA (short harpin RNA) from U6 or CMV promoter. shRNA gets processed by Drosha and Dicer RNAses inside the cell before it translocates to the cell cytoplasm and affects the level of target RNA. In this study we modified lentiviral vector pGIPZ expressing tFP-IRES-Puro-shRNA(mir30) cassette by introducing BamH I restriction site downstream of this cassette. This modification makes possible to clone specific shRNA sequences in pGIPZ vector using XhoI/BamHI restriction sites instead of the original recombination. Three shRNAs against phosphoprotein P of respiratory sinthitial virus (RSV) and shRNA against human CD43 as a control were generated and cloned into modified so-called pCIPD vector. Monkey kidney cells MA-104 were stably transduced with four shRNA constructs. In conclusion, the generated lentiviral vector pCIPD can be successfully used for efficient gene silencing and virus replication in a broad variety of cells.

Asunto(s)

Genes Virales , Vectores Genéticos , Lentivirus , Interferencia de ARN , Virus Sincitiales Respiratorios/genética , Virus Sincitiales Respiratorios/metabolismo , Animales , Línea Celular , Humanos , Macaca

RESUMEN

Two fragments of conjugative plasmid p19 (95 kb) from the soil strain Bacillus subtilis 19 were cloned and sequenced; these fragments carry genes, products of which are indispensable for the conjugative transfer. One of the fragments 4518 bp in size carries five open reading frames and their fragments (ORF1-ORF5). The protein corresponding to ORF4 is homologous to proteins from the family VirD4. Inactivation of ORF4 and ORF1 by insertional mutagenesis caused a three-to-fivefold decrease in the frequency of plasmid p19 conjugative transfer. Another 2932-bp fragment of p19 was shown to possess a rep region homologous to the rep region of plasmid pBS72 from the soil strain B. subtilis 72 and a novel ORF (ORF6); the protein corresponding to this ORF contains the HTH motif typical for DNA-binding proteins.

Asunto(s)

Bacillus subtilis/genética , Conjugación Genética , Genes Bacterianos , Plásmidos , Microbiología del Suelo

RESUMEN

A conjugal retrotransfer-retromobilization of a small nonconjugative plasmid pUB110 was established in Bacillus subtilis. This process involves a large conjugative plasmid p19.

Asunto(s)

Bacillus subtilis/genética , Conjugación Genética , Plásmidos/genética , ADN Bacteriano/genética