RESUMEN
Rising healthcare for elder and disabled people can be controlled by offering people autonomy at home by means of information technology. In this paper, we present an original and sensorless alert management solution which performs multimedia and home automation service discrimination and extracts highly regular home activities as sensors for alert management. The results of simulation data, based on real context, allow us to evaluate our approach before application to real data.
Asunto(s)
Servicios de Salud para Ancianos , Monitoreo Ambulatorio/instrumentación , Anciano , Algoritmos , Automatización , Análisis por Conglomerados , Personas con Discapacidad , Electrónica Médica , Diseño de Equipo , Servicios de Atención de Salud a Domicilio , Humanos , Modelos Estadísticos , Monitoreo Ambulatorio/métodos , Programas Informáticos , Factores de TiempoRESUMEN
Expanded bed absorption chromatography (EBA) was used to improve and simplify the purification of several wheat recombinant proteins. Binding and elution conditions were set to allow the purification of the over expressed protein in a single step. In comparison with our previous multi step protocol, same purity was obtained while EBA required less time (one day instead of five) and gave a higher yield (63% instead of 10%). This new procedure was then used for the successful purification of five other wheat ns-LTP. Despite their important polymorphism (identity from 44 to 97 %-pHi from 8 to 10), the EBA protocol allowed their purification in a single step.
Asunto(s)
Cromatografía de Afinidad/métodos , Proteínas de Plantas/aislamiento & purificación , Proteínas Recombinantes/aislamiento & purificación , Triticum/metabolismo , Adsorción , Proteínas de la Membrana/aislamiento & purificación , Proteínas de Transferencia de Fosfolípidos/aislamiento & purificación , Pichia/metabolismoRESUMEN
The thioredoxin h system has the specific capability to reduce intramolecular disulfide bonds of proteins, thereby modifying their tertiary structure. It is involved in many processes: in the activation or deactivation of enzymes and enzyme inhibitors and in the germination process. This system can be used to improve the breadmaking quality of wheat by strengthening the dough. It can also decrease the epitope accessibility, then modifying the response of the IgE immune system. Transgenic barley and wheat have been created to confirm the functionality of the NADP-dependent thioredoxin h system.
Asunto(s)
Grano Comestible/genética , Industria de Alimentos/tendencias , Germinación/fisiología , Plantas Modificadas Genéticamente , Tiorredoxinas/metabolismo , Alérgenos/química , Alérgenos/inmunología , Pan , Grano Comestible/metabolismo , Hordeum/genética , NADP/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/inmunología , Tiorredoxina h , Tiorredoxinas/genética , Triticum/genéticaRESUMEN
Postgenomic studies have led to an increasing demand for isotope-labeled proteins. We present a method for producing large quantities of truly native (15)N-labeled protein. Based on the secretion capabilities of the yeast Pichia pastoris, the recombinant protein is easily purified in a single step as it is secreted. Control of all nitrogen sources permits very high labeling yields. As a result, accumulation and folding of the recombinant protein can be monitored by heteronuclear NMR without purification. Comparison of sample spectra with the spectrum of the purified recombinant protein allows detection of the secreted protein in the culture and monitoring of its folding, from the start of the induction phase. The detection limit for a (15)N-labeled protein is estimated as 20 microM and corresponds, for a 10-kDa protein, to a load of 40 mg/liter in the fermentor. This concentration is reached by most reported preparations in P. pastoris. Further concentration by ultrafiltration would compensate for lower production. This procedure may be useful in many structural genomics and combinatorial chemistry screening projects where most protein productions meet the requirements for this method.
Asunto(s)
Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Pichia/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Pliegue de Proteína , Triticum/genética , Antígenos de Plantas , Proteínas Portadoras/biosíntesis , Técnicas Químicas Combinatorias/métodos , Peso Molecular , Isótopos de Nitrógeno , Resonancia Magnética Nuclear Biomolecular/métodos , Pichia/metabolismo , Proteínas de Plantas/biosíntesis , Protones , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/aislamiento & purificación , Triticum/químicaRESUMEN
Durum wheat (Triticum durum, Desf.) endosperm of mature kernels contained a single form of glutathione reductase (GR); it appeared about the 18th day after anthesis while another isoform, present at the early stages of grain development, disappeared between the 20th and 30th days after flowering. The form that was present at grain maturity was isolated and characterized. It was composed of two monomers, each one having an apparent molecular mass of about 60 kDa. The K(m) values for NADPH and for GSSG were 3.7 and 9.1 microM, respectively, and the V(m) values for NADPH and for GSSG were 594 and 575 microkat.mg(-)(1) protein, respectively. The pH(i) of the enzyme was situated between pH 4.4 and 4.5. At a constant temperature of 25 degrees C, the optimum GR activity was found to be between pH 7.5 and 8.0. It was relatively resistant to high temperatures and was very resistant to very low temperatures.
Asunto(s)
Glutatión Reductasa/química , Glutatión Reductasa/aislamiento & purificación , Triticum/enzimología , Western Blotting , Electroforesis en Gel de Poliacrilamida , Concentración de Iones de Hidrógeno , Peso Molecular , Proteínas de Plantas/química , Proteínas de Plantas/aislamiento & purificaciónRESUMEN
Remodelling of bone, regularly observed after the implantation of a hydroxyapatite coated Furlong hip prosthesis, stimulated the authors to analyse the stress distribution and relative magnitude in photoelastic models, of 1. the upper half of an intact femur; 2. of a femur equipped with a prosthesis free of glue to simulate the implantation of a cementless non coated prosthesis; and 3. of a femur in which a prosthesis was glued to simulate the implantation of a hydroxyapatite coated prosthesis. The results match precisely the remodelling thus observed in the X-rays.
Asunto(s)
Remodelación Ósea/fisiología , Prótesis de Cadera , Modelos Biológicos , Fenómenos Biomecánicos , Elasticidad , Humanos , Hidroxiapatitas , Diseño de Prótesis , Estrés Mecánico , Soporte de PesoRESUMEN
The reactivity of human thioredoxin (HTR) was tested in several reactions. HTR was as efficient as E. coli or plant and algal thioredoxins when assayed with E. coli ribonucleotide reductase or for the reduction of insulin. On the other hand, HTR was poorly reduced by NADPH and the E. coli flavoenzyme NADPH thioredoxin reductase as monitored in the DTNB reduction test. When reduced with dithiothreitol (DTT), HTR was much less efficient than thioredoxin m and thioredoxin f, the respective specific thioredoxins for the chloroplast enzymes NADP-malate dehydrogenase (NADP-MDH) and fructose 1,6 bisphosphatase (FBPase). Finally, HTR could be used in the photoactivation of NADP-MDH although less efficiently than thioredoxin m, proving nevertheless that it can be reduced by the iron sulfur enzyme ferredoxin thioredoxin reductase in the presence of photoreduced ferredoxin. Based on sequence comparisons, it was expected that HTR would display a reactivity similar to chloroplast thioredoxin f rather than to thioredoxin m. However the observed behavior of FTR did not exactly fit this prediction. The results are discussed in relation to the structural data available for the proteins.
Asunto(s)
Fructosa-Bifosfatasa/metabolismo , Oxidorreductasas/metabolismo , Tiorredoxinas/farmacología , Secuencia de Aminoácidos , Tiorredoxinas en Cloroplasto , Cloroplastos/efectos de los fármacos , Cloroplastos/enzimología , Ditiotreitol/farmacología , Activación Enzimática/efectos de los fármacos , Escherichia coli/enzimología , Fructosa-Bifosfatasa/química , Humanos , Malato Deshidrogenasa/química , Malato Deshidrogenasa/metabolismo , Malato-Deshidrogenasa (NADP+) , Datos de Secuencia Molecular , NADP/farmacología , Oxidorreductasas/química , Ribonucleótido Reductasas/química , Ribonucleótido Reductasas/metabolismo , Relación Estructura-Actividad , Reductasa de Tiorredoxina-Disulfuro/química , Reductasa de Tiorredoxina-Disulfuro/metabolismo , Tiorredoxinas/químicaRESUMEN
Photoelastic studies were performed to compare the effects of meniscectomy, longitudinal meniscal tear, and retention of the meniscal rim on the magnitude and distribution of the stresses in the medial compartment of the knee. The results are consistent with the concepts that the meniscus protects the articular cartilage of the knee from stress concentrations and that retention of a torn, undisplaced meniscus or the presence of an outer meniscal rim are preferable to the absence of the meniscus.