RESUMEN
Purpose: To assess the occurrence and diagnostic performance of nine single-nucleotide variants (SNVs) in the TCF4, SLC4A11, LOXHD1, and AGBL1 genes and the CTG18.1 trinucleotide repeat expansion in a Russian cohort of Fuchs' endothelial corneal dystrophy (FECD) patients. Methods: This retrospective case-control study included 100 patients diagnosed with FECD (cases) and 100 patients with cataracts (controls). Blood DNA was used to perform PCR and subsequent Sanger sequencing of rs613872 and rs17595731 in TCF4, c.99-100delTC, rs267607065, rs267607064, and rs267607066 in SLC4A11, rs113444922 in LOXHD1, and rs181958589 and rs185919705 in AGBL1. The number of CTG18.1 trinucleotide repeats was determined by a combination of conventional PCR or triplet primed PCR with fragment analysis. Results: At least one rs613872 marker allele was found in 78% of FECD patients and 21% of controls, and at least one rs17595731 marker allele was found in 14% and 2%, respectively. CTG18.1 trinucleotide expansion (>40 repeats) was detected in 72% of FECD patients and 5% of controls. Marker alleles of the tested SNVs in SLC4A11, LOXHD1, and rs185919705 in AGBL1 were not found in our FECD cohort. One FECD patient carried the marker allele of the rs181958589 SNV. Analysis of the diagnostic performance of individual markers in TCF4 and their combinations showed that the CTG18.1 repeat expansion was the best classifier for FECD (AUC = 0.84). Conclusions: Patients carrying CTG18.1 repeat expansion constituted a high proportion of the Russian FECD cohort; therefore, this marker is suitable for development of diagnostic and therapeutic approaches.
Asunto(s)
Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/genética , Distrofia Endotelial de Fuchs/genética , Marcadores Genéticos/genética , Expansión de Repetición de Trinucleótido/genética , Anciano , Anciano de 80 o más Años , Proteínas de Transporte de Anión/genética , Antiportadores/genética , Proteínas Portadoras/genética , Estudios de Casos y Controles , Femenino , Distrofia Endotelial de Fuchs/etnología , Técnicas de Genotipaje , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Polimorfismo de Nucleótido Simple , Estudios Retrospectivos , Federación de Rusia/epidemiología , Factor de Transcripción 4/genética , Población Blanca/etnologíaRESUMEN
The Mycoplasma hominis vaa gene encodes a highly variable surface antigen involved in adhesion to host cells. We studied 15 clinical isolates of Mycoplasma hominis with three types of the vaa gene. These vaa versions determine various forms of Vaa protein, which are characterized by different quantity and structure of homologous replaceable cassettes. Each cassette contains heptad repeats and sites for adherence. The differences on single nucleotides were observed in the primary sequences of the homologous modules of the vaa gene. A high frequency of nucleotide replacements in V module of the vaa gene (first and/or second position in codon) was determined. This region with various clusters of direct and indirect repeats of nucleotide sequences is incorporated into the area of the vaa gene. Amino-acid sequences corresponding to the hyper-variable region of the vaa gene are associated with the sections of coiled-coils and loops of Vaa. These bacterial regions involved in interaction with the host cell membranes could yield useful indications for more insights into the mechanism of mycoplasma persistence in humans.
Asunto(s)
Adhesinas Bacterianas/genética , Variación Genética , Infecciones por Mycoplasma/microbiología , Mycoplasma hominis/genética , Variación Antigénica , ADN Bacteriano/química , ADN Bacteriano/genética , Humanos , Modelos Moleculares , Mycoplasma hominis/química , Mycoplasma hominis/aislamiento & purificación , Estructura Secundaria de Proteína , Estructura Terciaria de Proteína , Secuencias Repetitivas de Ácidos Nucleicos , Análisis de Secuencia de ADNRESUMEN
The in vivo action of the antimicrobial peptide melittin, expressed from a recombinant plasmid vector, on chickens experimentally infected with Mycoplasma gallisepticum was studied. The plasmid vector pBI/mel2/rtTA includes the melittin gene under the control of an inducible tetracycline-dependent human cytomegalovirus promoter and the gene coding for the trans-activation protein rtTA. Aerosol administration of the vector, followed by infecting the chickens with M. gallisepticum 1226, is shown to inhibit development of infection. The inhibitory action was confirmed by a complex of clinical, pathomorphological, histological and serological studies, and also by comparing the M. gallisepticum reisolation frequency from the respiratory tract and internal organs. The data suggest that plasmid vectors expressing genes of antimicrobial peptides can be considered as potential agents for the prevention and treatment of mycoplasma infections in poultry farming.