This paper describes a subculture system of rice calli (cultivar J-104) that maintains the potential for long-term regeneration. We examined the effects of 2.4-D (1.0, 2.0, 3.0, 4.0 and 5.0 mg/L) and time of subculture on the morphogenesis and regeneration potential. Calli were subcultured for two years on semisolid medium, in the dark. During this time, calli with similar weight were monthly transferred to the regeneration medium, under photoperiod regime. Plant regeneration took place through two morphogenic pathways: indirect somatic embryogenesis and indirect organogenesis. The concentration of 2.4-D used in the callogenesis determined the prevalence of one path or another. In the first ten months of subculture, embryogenesis prevailed over organogenesis. However, from this point to the final stages, the shoots were organogenic. Callus lines retained their regeneration potential by organogenesis until the end of the second year, without presenting a significant number of abnormal plants. These results indicate that it is possible to maintain for two years J-104 rice callus lines potentially morphoge-nic, which can be applied to a plastid transformation protocol, specifically to the design of an efficient and long-stage selection, for reaching of homoplasmy during the callus period, before the regeneration of the plants.

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<p>Se analizó la expresión constitutiva del gen reportero de la ß-Glucuronidasa (GUS) fusionado a tres promotores: el 35S del virus del mosaico de la coliflor (CaMV), el promotor quimérico A9 que contiene la actina-1 de arroz y el promotor ubiquitina-1 de maíz. La actividad de los promotores fue analizada cualitativa y cuantitativamente en diferentes tejidos y estadíos de crecimiento de plantas de arroz (variedad J-104) transformadas mediante biobalística. Se demostró la expresión constitutiva de GUS bajo los promotores estudiados, con distintos patrones de actividad relativa en hojas, tallos y raíces de plantas in vitro y ex vitro, y en plantas de la progenie T 1. Bajo el promotor quimérico A9 se lograron los mayores niveles de expresión GUS en todos los tejidos y fases de crecimiento de las plantas.</p>

This work describes the validation of an enzyme-linked immunosorbent assay (ELISA) for detection of anti-Saccharomyces cerevisiae antibodies (ASCA) in diabetic patients with foot ulcers, after the treatment with Heberprot-P®. Validation followed regulatory guidelines of US FDA and European Medicine Agency. Minimum required dilution of samples and quality controls were defined using pools of sera from diabetic patients and from healthy donors. Parameters such as cut point, specificity, precision, selectivity, robustness and sample stability were analyzed. The repeatability and intermediate precision percent ranged between 7.93-10.61% and 7.93-11.43 %, respectively, indicating low intra- and inter-assay variation. The specificity was proved by background noise suppression, reaching 100% of inhibition as strong criterion for the specificity of the immunoassay. The validated ELISA is a reliable tool for ASCA detection in human serum after the administration of Heberprot-P®, in order to find immunological reactions associated with latent contamination by host cell proteins from Saccharomyces cerevisiae.