RESUMEN
A multijet and multistage aerosol concentrator was designed and fabricated with two virtual impactors in a series. Collection efficiency, internal loss, and concentration factors were calculated at ambient conditions for each stage. The total inlet flow rate of the aerosol concentrator was set at 1000 L/min(-1), while the minor flow rate for the first stage was at 6.0% of the total inlet flow and the minor flow rate of the second stage was at 6.7% of the first stage minor flow. The aerosol concentrator was calibrated using polystyrene latex particles in aerodynamic sizes ranging from 0.5 to 10 microm. Several configurations of the multijet acceleration nozzles and multitube receptors were designed in this study. The effects of the different designs were subsequently evaluated through experimentation. It was found that a properly designed multijet and multistage aerosol concentrator can significantly improve aerosol concentration performance. Results showed that the concentration factor increases from 1 to 240 over the particle size range studied. Applications of the multijet and multistage aerosol concentrator with high-volume flow rate can vary widely, from detection of biological aerosols at low concentration, laboratory aerosol sampling, clean room monitoring, and ambient aerosol measurements.
Asunto(s)
Aerosoles , Evaluación Preclínica de Medicamentos/instrumentación , Monitoreo de Drogas/instrumentación , Monitoreo del Ambiente/instrumentación , Movimientos del Aire , Evaluación Preclínica de Medicamentos/normas , Monitoreo de Drogas/normas , Monitoreo del Ambiente/normas , Diseño de Equipo , Humanos , Ensayo de Materiales , Tamaño de la Partícula , Distribución TisularRESUMEN
Transforming growth factor beta (TGF-beta) has been considered as a candidate for gene therapy of orthopedic diseases. The possible application of cell-mediated TGF-beta gene therapy as a new treatment regimen for degenerative arthritis was investigated. In this study, fibroblasts expressing active TGF-beta 1 were injected into the knee joints of rabbits with artificially made cartilage defects to evaluate the feasibility of this therapy for orthopedic diseases. Two to 3 weeks after the injection there was evidence of cartilage regeneration, and at 4 to 6 weeks the cartilage defect was completely filled with newly grown hyaline cartilage. Histological analyses of the regenerated cartilage suggested that it was well integrated with the adjacent normal cartilage at the sides of the defect and that the newly formed tissue was indeed hyaline cartilage. Our findings suggest that cell-mediated TGF-beta 1 gene therapy may be a novel treatment for orthopedic diseases in which hyaline cartilage damage has occurred.
Asunto(s)
Cartílago/química , Fibroblastos/metabolismo , Terapia Genética/métodos , Hialina/metabolismo , Factor de Crecimiento Transformador beta/biosíntesis , Células 3T3 , Animales , Artritis/metabolismo , Northern Blotting , Cartílago/metabolismo , Diferenciación Celular , División Celular , Condrocitos/metabolismo , ADN Complementario/metabolismo , Vectores Genéticos/genética , Inmunohistoquímica , Imagen por Resonancia Magnética , Ratones , Unión Proteica , ARN Mensajero/metabolismo , Conejos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor de Crecimiento Transformador beta1 , TransgenesRESUMEN
S-Adenosyl-L-methionine (SAM): coclaurine N-methyltransferase (CNMT), which catalyzes the transfer of a methyl group from S-adenosyl-L-methionine to the amino group of the tetrahydrobenzylisoquinoline alkaloid coclaurine. was purified 340-fold from Coptis japonica cells in 1% yield to give an almost homogeneous protein. The purified enzyme, which occurred as a homotetramer with a native Mr of 160 kDa (gel-filtration chromatography) and a subunit Mr of 45 kDa (SDS-polyacrylamide gel electrophoresis), had an optimum pH of 7.0 and a pI of 4.2. Whereas (R)-coclaurine was the best substrate for enzyme activity, Coptis CNMT had broad substrate specificity and no stereospecificity CNMT methylated norlaudanosoline, 6,7-dimethoxyl-1,2,3,4-tetrahydroisoquinoline and 1-methyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline. The enzyme did not require any metal ion. p-Chloromercuribenzoate and iodoacetamide did not inhibit CNMT activity, but the addition of Co2+, Cu2+ or Mn2+ at 5 mM severely inhibited such activity by 75, 47 and 57%, respectively. The substrate-saturation kinetics of CNMT for norreticuline and SAM were of the typical Michaelis-Menten-type with respective Km values of 0.38 and 0.65 mM.