RESUMO
Cecropin A is a natural antimicrobial peptide that exhibits fast and potent activity against a broad spectrum of pathogens and neoplastic cells, and that has important biotechnological applications. However, cecropin A exploitation, as for other antimicrobial peptides, is limited by their production and purification costs. Here, we report the efficient production of this bioactive peptide in rice bran using the rice oleosin 18 as a carrier protein. High cecropin A levels were reached in rice seeds driving the expression of the chimeric gene by the strong embryo-specific oleosin 18 own promoter, and targeting the peptide to the oil body organelle as an oleosin 18-cecropin A fusion protein. The accumulation of cecropin A in oil bodies had no deleterious effects on seed viability and seedling growth, as well as on seed yield. We also show that biologically active cecropin A can be easily purified from the transgenic rice seeds by homogenization and simple flotation centrifugation methods. Our results demonstrate that the oleosin fusion technology is suitable for the production of cecropin A in rice seeds, which can potentially be extended to other antimicrobial peptides to assist their exploitation.
Assuntos
Peptídeos Catiônicos Antimicrobianos/biossíntese , Gotículas Lipídicas/química , Oryza/metabolismo , Sementes/metabolismo , Sequência de Aminoácidos , Peptídeos Catiônicos Antimicrobianos/genética , Genoma de Planta , Espectrometria de Massas , Dados de Sequência Molecular , Oryza/genética , Fenótipo , Óleos de Plantas/química , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Regiões Promotoras Genéticas , Proteínas Recombinantes de Fusão/metabolismo , Sementes/genética , TransgenesRESUMO
OBJETIVO: Realizar una revisión bibliográfica para identificar las diferentes metodologías empleadas en el proceso de conciliación de los tratamientos farmacológicos que sean aplicables a pacientes pluripatológicos. DISEÑO: Revisión sistemática. Fuentes de datos: Se realizó una revisión bibliográfica (febrero de 2012) en las bases de datos Pubmed, EMBASE, CINAHL, PsycINFO e Índice Médico Español de métodos de conciliación del tratamiento en pacientes pluripatológicos, o en su defecto, ancianos o polimedicados. Selección de estudios: Se recuperaron 273 artículos, de los que se seleccionaron 25. Extracción de datos: Se extrajo información relativa a la metodología empleada: nivel asistencial en el que se realiza, fuentes de información, uso de formulario, tiempo establecido, profesional responsable, información recogida y variables registradas como errores de conciliación. RESULTADOS: La mayoría de estudios fueron al ingreso y al alta hospitalarios Como principales fuentes de información destacan la entrevista y la historia clínica. En la mayoría de trabajos no se especifica un tiempo preestablecido, ni se usa formulario, y el principal responsable es el farmacéutico clínico. Además de la medicación domiciliaria, los hábitos de automedicación y la fitoterapia también son registrados. Se recogen como errores de conciliación desde omisiones de fármacos hasta interacciones medicamentosas. CONCLUSIONES: Existe gran heterogeneidad en la metodología empleada para la actividad de la conciliación. No existe ningún trabajo realizado específicamente en el paciente pluripatológico, que por su complejidad y susceptibilidad a errores de conciliación requiere una metodología estandarizada
OBJECTIVE: To carry out a ibliographic review in order to identify the different methodologies used along the reconciliation process of drug therapy applicable to polypathological patients. DESIGN: We performed a literature review. Data sources: The bibliographic review (February 2012) included the following databases: Pubmed, EMBASE, CINAHL, PsycINFO and Spanish Medical Index (IME). The different methodologies, identified on those databases, to measure the conciliation process in polypathological patients, or otherwise elderly patients or polypharmacy, were studied. Study selection Two hundred and seventy three articles were retrieved, of which 25 were selected. Data extraction Specifically: the level of care, the sources of information, the use of registration forms, the established time, the medical professional in charge and the registered variables such as errors of reconciliation. RESULTS: Most of studies selected when the patient was admitted into the hospital and after the hospital discharge of the patient. The main sources of information to be highlighted are: the interview and the medical history of the patient. An established time is not explicitly stated on most of them, nor the registration form is used. The main professional in charge is the clinical pharmacologist. Apart from the home medication, the habits of self-medication and phytotherapy are also identified. The common errors of reconciliation vary from the omission of drugs to different forms of interaction with other medicinal products (drugs interactions). CONCLUSIONS: There is a large heterogeneity of methodologies used for reconciliation. There is not any work done on the specific figure of the polypathological patient, which precisely requires a standardized methodology due to its complexity and its susceptibility to errors of reconciliation
Assuntos
Humanos , Quimioterapia Combinada , Reconciliação de Medicamentos/métodos , Doença Crônica/tratamento farmacológico , Coleta de Dados , Comorbidade , Doença Crônica/epidemiologiaRESUMO
OBJECTIVE: To carry out a bibliographic review in order to identify the different methodologies used along the reconciliation process of drug therapy applicable to polypathological patients. DESIGN: We performed a literature review. Data sources The bibliographic review (February 2012) included the following databases: Pubmed, EMBASE, CINAHL, PsycINFO and Spanish Medical Index (IME). The different methodologies, identified on those databases, to measure the conciliation process in polypathological patients, or otherwise elderly patients or polypharmacy, were studied. Study selection Two hundred and seventy three articles were retrieved, of which 25 were selected. Data extraction Specifically: the level of care, the sources of information, the use of registration forms, the established time, the medical professional in charge and the registered variables such as errors of reconciliation. RESULTS: Most of studies selected when the patient was admitted into the hospital and after the hospital discharge of the patient. The main sources of information to be highlighted are: the interview and the medical history of the patient. An established time is not explicitly stated on most of them, nor the registration form is used. The main professional in charge is the clinical pharmacologist. Apart from the home medication, the habits of self-medication and phytotherapy are also identified. The common errors of reconciliation vary from the omission of drugs to different forms of interaction with other medicinal products (drugs interactions). CONCLUSIONS: There is a large heterogeneity of methodologies used for reconciliation. There is not any work done on the specific figure of the polypathological patient, which precisely requires a standardized methodology due to its complexity and its susceptibility to errors of reconciliation.
Assuntos
Reconciliação de Medicamentos/métodos , HumanosRESUMO
The Aspergillus giganteus antifungal protein (AFP), encoded by the afp gene, has been reported to possess in vitro antifungal activity against various economically important fungal pathogens, including the rice blast fungus Magnaporthe grisea. In this study, transgenic rice ( Oryza sativa ) constitutively expressing the afp gene was generated by Agrobacterium -mediated transformation. Two different DNA constructs containing either the afp cDNA sequence from Aspergillus or a chemically synthesized codon-optimized afp gene were introduced into rice plants. In both cases, the DNA region encoding the signal sequence from the tobacco AP24 gene was N-terminally fused to the coding sequence of the mature AFP protein. Transgenic rice plants showed stable integration and inheritance of the transgene. No effect on plant morphology was observed in the afp -expressing rice lines. The inhibitory activity of protein extracts prepared from leaves of afp plants on the in vitro growth of M. grisea indicated that the AFP protein produced by the trangenic rice plants was biologically active. Several of the T(2) homozygous afp lines were challenged with M. grisea in a detached leaf infection assay. Transformants exhibited resistance to rice blast at various levels. Altogether, the results presented here indicate that AFP can be functionally expressed in rice plants for protection against the rice blast fungus M. grisea.