RESUMEN
The article presents the developed and approved test-systems for evaluation of the level of expression of immune system responsible activation and inhibition of T-cell response in recipients of renal transplant under application of extra-corporal photochemotherapy.
Asunto(s)
Trasplante de Riñón , Fotoquimioterapia , Linfocitos TRESUMEN
Nucleic acid-based therapeutics have recently emerged as a new class of next generation agents for treatment and prevention of viral infection, cancer, and genetic disorders, but their wide use is limited by their relatively weak delivery into target cells. Usage of synthetic cationic amphiphiles with peptide hydrophilic domain as agents for non-viral gene delivery is an attractive approach. We developed the schemes for the synthesis of aliphatic peptides with different length of the hydrocarbon chains in hydrophobic domains and different amino acids in polar head. For the obtained derivatives we determined transfection efficiency, critical vesicle concentration, particle size, ζ-potential and aggregates stability. We have found that the transfection efficiency is increased if the ornithine is a part of polar head in an amphiphile. The most promising amphiphile for liposomal formation OrnOrnGlu(C16H33)2 was examined more carefully. It has been shown that the lipopeptide possesses low toxicity (in vitro and in vivo) and high transfection efficiency with pDNA and siRNA in different cell lines. In addition, the production of liposomes based on this lipopeptide is simple, quick and cheap. Thus OrnOrnGlu(C16H33)2 is a promising vehicle for gene delivery and gene silencing.
Asunto(s)
Silenciador del Gen/efectos de los fármacos , Lipopéptidos/administración & dosificación , Lipopéptidos/química , Células A549 , Animales , Células CHO , Cationes/administración & dosificación , Cationes/química , Línea Celular , Línea Celular Tumoral , Cricetulus , ADN/metabolismo , Técnicas de Transferencia de Gen , Terapia Genética/métodos , Células HEK293 , Células HeLa , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Células Jurkat , Liposomas/administración & dosificación , Liposomas/química , Tamaño de la Partícula , Plásmidos/administración & dosificación , Plásmidos/química , ARN Interferente Pequeño/metabolismo , Transfección/métodosRESUMEN
Comparative evaluation of efficiency of several methods of DNA extraction from storage cultures of acidophilic chemolithotrophic microorganism communities isolated from sulfide ores of Shanuch ore deposit (Kamchatka peninsula) was conducted. DNA extraction methods in various combinations of physical (heating to 65-98 degrees C, grinding with SiO2 particles), enzymatic (treatment with lysozyme and proteinase K), and chemical (GuSCN, CTAB and KOH) treatments were tested. The evaluation of efficiency was performed using Real-time PCR. The best result was obtained for the combined method based on GuSCN lysis activity (lysis at 65 degrees C) followed by purification with phenol and chloroform.
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ADN de Archaea/aislamiento & purificación , ADN Bacteriano/aislamiento & purificación , Minerales/química , Cetrimonio , Compuestos de Cetrimonio/química , Crecimiento Quimioautotrófico/fisiología , Cloroformo/química , Endopeptidasa K/química , Industria Procesadora y de Extracción , Asia Oriental , Calor , Consorcios Microbianos/fisiología , Muramidasa/química , Fenol/química , Reacción en Cadena en Tiempo Real de la Polimerasa , Federación de Rusia , Dióxido de Silicio/químicaAsunto(s)
Isquemia Miocárdica/complicaciones , Trombosis/etiología , Enfermedad Crónica , Femenino , Estudios de Seguimiento , Humanos , Incidencia , Masculino , Persona de Mediana Edad , Isquemia Miocárdica/diagnóstico , Prevalencia , Estudios Prospectivos , Factores de Riesgo , Federación de Rusia/epidemiología , Trombosis/epidemiologíaAsunto(s)
Quimiocina CXCL12/genética , Infecciones por VIH/genética , Polimorfismo Genético , Receptores CCR2/genética , Receptores CCR5/genética , Síndrome de Inmunodeficiencia Adquirida/genética , Síndrome de Inmunodeficiencia Adquirida/inmunología , Etnicidad/genética , Frecuencia de los Genes , Infecciones por VIH/inmunología , VIH-1 , Humanos , Inmunidad Innata/genética , Federación de Rusia/epidemiologíaRESUMEN
The existing diversity of the methods for detecting single nucleotide polymorphisms is so great that may perplex an unsophisticated researcher who chooses the appropriate molecular genetic toolkit. In this work, we tried to systematize and briefly describe the state-of-the-art methods for detecting oligonucleotide polymorphisms that are based on allele-specific PCR and hybridization with oligonucleotide probe as well as to characterize the methods considered with respect to their accuracy, cost, and simplicity.
Asunto(s)
Hibridación de Ácido Nucleico , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo de Nucleótido Simple , Alelos , Sondas de Oligonucleótidos/genéticaRESUMEN
The digestion of the milk sugar (lactose) is observed every normal child but not in every adult. The decreased lactase synthesis in some adults results in problems with digestion of the whole milk (primary hypolactasy). An association of lactase activity in adults with carrying of the allele T within the polymorphism C/T-13910 located upstream of the lactase gene and 100% association of hypolactasy with the genotype C/C has recently been shown for a Finnish sample. In the present work we determined the LCT* C/T_13910 genotypes and allele frequencies in populations from Russia. The genotype C/C frequencies varied from 36.6% for Russians to 88.2% for Chukchi and were close to the published medical and epidemiological data on hypolactasy frequencies in respective populations. Genotyping was performed by three different methods to identify the optimal one. Our results have shown that the studied locus is the key determinant for the primary hypolactasy development in various human populations. Consequently, the DNA diagnostics of the C/C genotype carrying is a promising predictive test to detect the primary hypolactasy long before its clinical development. Practical application of this type of diagnostics would be a step towards the individual-oriented medicine.