RESUMEN
Recent data suggest a possible relationship between cystic fibrosis (CF) pharmacotherapy, Aspergillus fumigatus colonization (AC) and/or allergic bronchopulmonary aspergillosis (ABPA). The aim of this study was to determine if anti-fungal defence mechanisms are influenced by CF pharmacotherapy, i.e. if (1) neutrophils form CF and non-CF donors differ in their ability to produce chitotriosidase (CHIT-1); (2) if incubation of isolated neutrophils with azithromycin, salbutamol, prednisolone or rhDNase might influence the CHIT-1 activity; and (3) if NETosis and neutrophil killing efficiency is influenced by rhDNase. Neutrophils were isolated from the blood of CF patients (n = 19; mean age 26·8 years or healthy, non-CF donors (n = 20; 38·7 years) and stimulated with phorbol-12-myristate-13-acetate (PMA), azithromycin, salbutamol, prednisolone or rhDNase. CHIT-1 enzyme activity was measured with a fluorescent substrate. NETosis was induced by PMA and neutrophil killing efficiency was assessed by a hyphae recovery assay. Neutrophil CHIT-1 activity was comparable in the presence or absence of PMA stimulation in both CF and non-CF donors. PMA stimulation and preincubation with rhDNase increased CHIT-1 activity in culture supernatants from non-CF and CF donors. However, this increase was significant in non-CF donors but not in CF patients (P < 0·05). RhDNase reduced the number of NETs in PMA-stimulated neutrophils and decreased the killing efficiency of leucocytes in our in-vitro model. Azithromycin, salbutamol or prednisolone had no effect on CHIT-1 activity. Stimulation of isolated leucocytes with PMA and treatment with rhDNase interfered with anti-fungal defence mechanisms. However, the impact of our findings for treatment in CF patients needs to be proved in a clinical cohort.
Asunto(s)
Fibrosis Quística/inmunología , Desoxirribonucleasas/uso terapéutico , Hexosaminidasas/metabolismo , Neutrófilos/enzimología , Neutrófilos/patología , Adolescente , Adulto , Anciano , Albuterol/farmacología , Albuterol/uso terapéutico , Aspergillus fumigatus/aislamiento & purificación , Azitromicina/farmacología , Azitromicina/uso terapéutico , Bacterias/aislamiento & purificación , Fibrosis Quística/tratamiento farmacológico , Fibrosis Quística/microbiología , Desoxirribonucleasas/genética , Trampas Extracelulares/efectos de los fármacos , Femenino , Hongos/aislamiento & purificación , Hexosaminidasas/análisis , Hexosaminidasas/biosíntesis , Humanos , Masculino , Persona de Mediana Edad , Neutrófilos/efectos de los fármacos , Neutrófilos/fisiología , Ésteres del Forbol/farmacología , Prednisolona/farmacología , Prednisolona/uso terapéutico , Esputo/microbiología , Adulto JovenAsunto(s)
Genética de Población/efectos de los fármacos , Mutación/efectos de los fármacos , Animales , Permeabilidad de la Membrana Celular/efectos de los fármacos , Aberraciones Cromosómicas , Electroforesis , Femenino , Genes Dominantes/efectos de los fármacos , Genes Letales/efectos de los fármacos , Técnicas Genéticas , Agencias Gubernamentales , Humanos , Agencias Internacionales , Concentración Máxima Admisible , Mutágenos , Mutación/efectos de la radiación , Efectos de la Radiación , Reproducción , Estadística como Asunto , Estados Unidos , United States Food and Drug AdministrationRESUMEN
A new plasma-based micropatterning technique, here referred to as plasma printing, combines the well known advantages given by the nonequilibrium character of a dielectric barrier discharge (DBD) and its operation inside small gas volumes with dimension between tens and hundreds of micrometres. The discharge is run at atmospheric pressure and can be easily implemented for patterned surface treatment with applications in biotechnology and microtechnology. In this work the local modification of dielectric substrates, e.g. polymeric films, is addressed with respect to coating and chemical functionalisation, immobilisation of biomolecules and area-selective electroless plating.