RESUMEN
Studies of two recent decades provided ample data on antibacterial peptides protecting animal and human epithelium of different types; however, no reports about the presence of these compounds in hair keratinocytes appeared up to the present time. Peptides were extracted from specimens of normal intact hair with citric acid solution in 50% ethanol. Antibacterial activity of the resultant extracts was evaluated on Candida albicans test culture by staining and microscopy, by evaluation of growth inhibition zones, and by inoculations. Direct contact with the extract destroyed the greater part of Candida albicans cells up to complete destruction of membranes. Application of the extract to dishes with agar with Candida albicans led in the formation of apparent zones of growth inhibition. The percentage of killed cells increased with prolongation of incubation with the extract. Electrophoresis of hair extract showed bands characteristic of antibacterial peptides RNase, psoriasin, and ß-defensins. Removal of the peptides from the extract by filtration through membrane filter led to loss of its activity. The results indicate that human hair keratinocytes possess congenital antibacterial immunity.
Asunto(s)
Antiinfecciosos/farmacología , Antifúngicos/farmacología , Candida albicans/efectos de los fármacos , Cabello/inmunología , Adolescente , Adulto , Antiinfecciosos/aislamiento & purificación , Niño , Femenino , Cabello/química , Humanos , Persona de Mediana EdadAsunto(s)
Hidrocarburo de Aril Hidroxilasas/metabolismo , Sistema Enzimático del Citocromo P-450/metabolismo , Epóxido Hidrolasas/metabolismo , Xenobióticos/metabolismo , Factores de Edad , Animales , Femenino , Hígado/enzimología , Pulmón/enzimología , Masculino , Ratas , Estaciones del Año , Factores SexualesRESUMEN
We developed a scheme of consecutive replacement of complex components of a known Brucella medium containing peptones and blood with simple analogs and created a synthetic medium for Helicobacter pylori culturing. H. pylori cells require hemic iron for their growth; an appreciable increment in biomass was ensured by hemoglobin, but not simpler hemocontaining compounds (hemin and cytochrome C). Glutamine (20 g/liter) was used as the main nitrogen-containing component, and other amino acids were added in trace amounts. Adhesion was provided by adding agarose gel (0.1%) also promoting the increase in biomass. The proposed medium of a certain chemical composition differs from the known foreign analogs by the presence of hemocontaining component (hemoglobin), short period of exponential growth, and appreciable accumulation of cell protein.
Asunto(s)
Técnicas Bacteriológicas , Medios de Cultivo/química , Helicobacter pylori/metabolismo , Animales , Sangre/metabolismo , Citocromos c/metabolismo , Helicobacter pylori/citología , Hemina/metabolismo , Hemoglobinas/metabolismo , HumanosRESUMEN
The presence of immunoglobulins to Malassezia spp. surface proteins in the sera from patients with atopic dermatitis and healthy subjects was studied. It was found that 28% of 25 examined patients with atopic dermatitis had IgE antibodies to Malassezia spp. surface protein preparation. All patients and 5 healthy subjects had IgG antibodies to this preparation. The presence and concentration of specific IgE antibodies in patients with atopic dermatitis correlated with reverse titers of IgG antibodies to this preparation (r=0.782). The medians of values reciprocal to IgG antibody titers in patients with atopic dermatitis with and without specific IgE antibodies to the preparation and in healthy subjects were 64, 1024, and 16, respectively. The preparation derived from Candida albicans (candidine) and previously derived preparation from Malassezia did not cross-react. According to immunoblotting data, the preparation contains allergens presented by proteins with molecular weights 15, 36, 52-56, and 78.4 kDa.
Asunto(s)
Extractos Celulares/inmunología , Dermatitis Atópica/inmunología , Inmunoglobulina E/inmunología , Inmunoglobulina G/inmunología , Malassezia/inmunología , Adolescente , Adulto , Extractos Celulares/química , Proteínas Fúngicas/inmunología , Humanos , Persona de Mediana Edad , Estadística como AsuntoRESUMEN
Malassezia spp. yeast habituating the skin of healthy humans can be a source of allergens for patients with atopic dermatitis. We proposed a method for obtaining allergen-containing preparation by trimming outer cell wall proteins with 1% sodium dodecyl sulfate. The resultant preparation contained 36 and 67 kD proteins known as Malassezia allergens. IgE antibodies to these proteins were detected in the sera of young people with atopic dermatitis.
Asunto(s)
Alérgenos/aislamiento & purificación , Antígenos Fúngicos/aislamiento & purificación , Proteínas Fúngicas/aislamiento & purificación , Malassezia/química , Adulto , Alérgenos/inmunología , Antígenos Fúngicos/inmunología , Pared Celular , Niño , Preescolar , Dermatitis Atópica/sangre , Dermatitis Atópica/inmunología , Dermatitis Atópica/microbiología , Proteínas Fúngicas/inmunología , Humanos , Malassezia/inmunología , Dodecil Sulfato de SodioRESUMEN
A fast, simple, and sensitive technique is described for the biochemical analysis of human MHC class I antigens. It is based on 2-D electrophoresis or 1-D isoelectric focusing, followed by immunoblotting. The need for radiolabelling of the cells is eliminated, and because whole cell lysates can be used the number of cells needed for analysis is reduced tenfold over conventional immunoprecipitation techniques. The method can also be used to analyze certain MHC class II antigens.