RESUMEN
Providing food with nutrition and functionality is crucial for sustaining human life. Rice (Oryza sativa L.) is a representative staple crop with high carbohydrate content but low amounts of essential amino acids, micronutrients, and carotenoids such as provitamin A. To improve the nutritional quality, rice endosperm was biofortified to accumulate carotenoids such as ß-carotene through genetic engineering (i.e., using synthetic carotenoid biosynthetic genes, a nonmammalian viral polycistronic sequence, and an optimized promoter and transit peptide) and high-throughput rice transformation (approximately 300 transgenic plants per construct). To facilitate the safety assessment of genetically modified food, molecular characterization was performed to select elite lines equipped with a single intergenic insertion of T-DNA, high transgene expression, in this case leading to high carotenoid content, and with phenotypic and compositional substantial equivalence. In this study, we present ß-carotene-biofortified rice event candidate lines eligible for commercial use and a disclosed molecular protocol for the development of biotech rice crops.
Asunto(s)
Oryza , Plantas Modificadas Genéticamente , beta Caroteno , Oryza/genética , Oryza/metabolismo , beta Caroteno/metabolismo , Plantas Modificadas Genéticamente/genética , Biofortificación , Ingeniería Genética/métodosRESUMEN
INTRODUCTION: Assessing patient expectations in orthopaedic surgery has gained significant importance over time. However, there have been only a few studies on how to measure such expectations in hand surgery. Against the backdrop, the study was designed to develop a valid and reliable expectations survey for patients undergoing hand surgery and to identify the correlations between preoperative expectations and postoperative satisfaction. MATERIALS AND METHODS: This is a three-phase prospective cohort study. In the first phase of the study (146 patients), patient expectations were assessed while developing a draft questionnaire based on frequency and clinical relevance. In the second phase (154 patients newly included), test-retest reliability was measured to ensure test consistency. The Intraclass Correlation Coefficient (ICC) served as a basis for developing the final survey questionnaire. In the third phase, we followed up with patients, who completed the preoperative expectations survey, 3 months after surgery to assess the fulfillment of their expectations. The Pearson correlation method was used to measure the association between preoperative expectations and postoperative satisfaction. RESULTS: In the first phase, 146 patients shared 406 different expectations, which were grouped into nine categories. Then, in the second phase, the final survey was populated by questionnaire items under respective category that have revealed strong test-retest reliability (ICC of 0.91). A significant positive correlation between patient expectations and satisfaction was observed (R = 0.181, p = 0.034). CONCLUSION: The survey was designed to offer a valid and reliable approach for the comprehensive assessment of patient expectations in hand surgery. The survey results show that patients with high expectations tend to be more satisfied with surgical outcomes. It is strongly believed that this approach would serve as a useful tool at a time when patient perspective is taken into account increasingly more in the clinical practice.
Asunto(s)
Mano , Motivación , Humanos , Reproducibilidad de los Resultados , Estudios Prospectivos , Mano/cirugía , Satisfacción del Paciente , Encuestas y CuestionariosRESUMEN
Foot-and-mouth disease virus (FMDV) 2A constructs have been successfully used for the production of "Golden Rice", a ß-carotene producing rice strain. However, to allay public fears and opposition to plants carrying a mammalian pathogenic viral sequence, 2A-like synthetic sequences from Thosea asigna virus and Infectious myonecrosis virus were used to coordinate the coexpression of carotenoid biosynthetic genes. Here, up to four carotenogenic genes encoding PSY, CRTI, BCH and BKT were concatenated and produced ß-carotene, zeaxanthin, and ketocarotenoids (astaxanthin and adonixanthin) in transgenic rice seeds displaying color variation due to the difference in carotenoid content and composition.
RESUMEN
CDR3 of the heavy-chain variable region of immunoglobulin is a region in which somatic mutation occurs heavily after secondary antibody response, resulting in an affinity maturation of antibodies in vivo. The aim of this study was to improve the affinity of a human single-chain variable fragment (scFv) specific for pre-S1 of hepatitis B virus (HBV) by introducing random mutagenesis in CDR3 variable region of heavy chain (V(H)) of the parental scFv clone 1E4. By using a BIAcore for panning and screening, we have selected three clones (A9, B2, and B9) with lower highest affinity (K(D)) than 1E4. Affinities of selected clones ranged from 1.7 x 10(7) mol/L to 6.3 x 10(8) mol/L, which were increased by factors of 1.4 to 4.0, respectively, compared to the parental clone. Binding inhibition assay using flow cytometry and polymerase chain reaction revealed that B2 (6.4 x 10(8) mol/L) had a higher neutralizing activity against pre-S1 or HBV virion binding to liver cell line. This anti-pre-S1 scFv can be considered as a potential therapeutic tool for a passive immunotherapy for HBV infection.
Asunto(s)
Anticuerpos contra la Hepatitis B/inmunología , Virus de la Hepatitis B/inmunología , Virus de la Hepatitis B/metabolismo , Región Variable de Inmunoglobulina/genética , Región Variable de Inmunoglobulina/inmunología , Mutación/genética , Biblioteca de Péptidos , Secuencia de Aminoácidos , Humanos , Inmunoglobulina G , Región Variable de Inmunoglobulina/química , Datos de Secuencia Molecular , Pruebas de Neutralización , Unión ProteicaRESUMEN
Transforming growth factor-beta1 (TGF-beta1) is a pleiotrophic cytokine that mediates differentiation, growth, and apoptosis. As a potent immunosuppressive agent, TGF-beta1 induces apoptosis in primary lymphocytes. However, it has been recognized that TGF-beta1 plays certain roles in development or progression of hematopoietic tumours via inhibition of Fas-mediated apoptotic cell death. Several studies have highlighted the mechanisms of TGF-beta1-induced Fas resistance and its contribution to aggressive tumour behavior. In this study, we have focused on the mechanisms by which TGF-beta1 protected leukaemia/lymphoma cells from Fas-mediated apoptosis. The presented study provides that TGF-beta1 inhibited Fas-mediated apoptosis of leukaemia/lymphoma cells in two distinct pathways. First, TGF-beta1 reduced expression of surface Fas receptors by blockade of trafficking cytoplasmic Fas to the surface, which allowed the leukaemia cells to resist Fas-mediated cell death. However, total Fas levels including both surface and cytoplasmic Fas were not altered, indicating that forced degradation of Fas or transcriptional regulation was not involved. Second, TGF-beta1 up-regulated Fas signaling pathway inhibitor cFLIPL to block the pro-caspase-8 cleavage and thus promoted survival of leukaemia/lymphoma cells. Our findings may partly explain why higher concentration of serum TGF-beta1 in cancer patient was related with poor prognosis.