A nonsteroidal glucocorticoid receptor antagonist.

Selective intracellular receptor antagonists are used clinically to ameliorate hormone-dependent disease states. Patients with Cushing's syndrome have high levels of the glucocorticoid, cortisol, and suffer significant consequences from this overexposure. High levels of this hormone are also implicated in exacerbating diabetes and the stress response. Selectively inhibiting this hormone may have clinical benefit in these disease states. To this end, we have identified the first selective, nonsteroidal glucocorticoid receptor (GR) antagonist. This compound is characterized by a tri-aryl methane core chemical structure. This GR-specific antagonist binds with nanomolar affinity to the GR and has no detectable binding affinity for the highly related receptors for mineralocorticoids, androgens, estrogens, and progestins. We demonstrate that this antagonist inhibits glucocorticoid-mediated transcriptional regulation. This compound binds competitively with steroids, likely occupying a similar site within the ligand-binding domain. Once bound, however, the compound fails to induce critical conformational changes in the receptor necessary for agonist activity.

Development and validation of an antibody-dependent cell-mediated cytotoxicity-reporter gene assay.

Humanized monoclonal antibodies (mAbs) are the fastest growing class of biological therapeutics that are being developed for various medical indications, and more than 30 mAbs are already approved and in the market place. Antibody-dependent cell-mediated cytotoxicity (ADCC) is an important biological function attributed to the mechanism of action of several therapeutic antibodies, particularly oncology targeting mAbs. The ADCC assay is a complicated and highly variable assay. Thus, the use of an ADCC assay as a lot release test or a stability test for clinical trial batches of mAbs has been a substantial challenge to install in quality control laboratories. We describe here the development and validation of an alternate approach, an ADCC-reporter gene assay that is based on the key attributes of the PBMC-based ADCC assay. We tested the biological relevance of this assay using an anti-CD20 based model and demonstrated that this ADCC-reporter assay correlated well with standard ADCC assays when induced with the drugable human isotypes [IgG1, IgG2, IgG4, IgG4S > P (S228P) and IgG4PAA (S228P, F234A, L235A)] and with IgG1 isotype variants with varying amounts of fucosylation. This data demonstrates that the ADCC-reporter gene assay has performance characteristics (accuracy, precision and robustness) to be used not only as a potency assay for lot release and stability testing for antibody therapeutics, but also as a key assay for the characterization and process development of therapeutic molecules.

Concentrado proteico do soro melhora a qualidade sensorial de sobremesa láctea diet / Whey protein concentrate improves the sensory quality of free-fat dairy desserts

Alimentos isentos ou de baixo nível de gordura constituem um desafio para a indústria de alimentos. Nesse contexto, este trabalho teve como objetivo estudar o efeito da adição de concentrado proteico de soro (CPS) nas características sensoriais e aceitabilidade de sobremesas lácteas sem gordura. Quatro formulações foram desenvolvidas com diferentes concentrações de CPS (0%, 1,5%, 3,0% e 4,5% m v-1) e caracterizadas sensorialmente por meio da Análise Descritiva. A aceitabilidade das formulações foi avaliada por 94 consumidores que utilizaram escala hedônica de nove pontos. As sobremesas foram caracterizadas por oito atributos sensoriais: cor, firmeza, resistência ao corte, brilho, consistência, gomosidade, aroma e sabor de baunilha. As formulações contendo 3,0 e 4,5% de CPS apresentaram maior intensidade de todos os atributos sensoriais. A adição de CPS foi efetiva na composição da textura das sobremesas lácteas diet, além de realçar a cor, sabor e aroma do produto. As formulações contendo 1,5 e 3,0% de CPS foram as mais aceitas, evidenciando que a utilização do CPS nessas concentrações favoreceu a aceitabilidade do produto, proporcionando características sensoriais agradáveis aos consumidores.

Foods free or low-fat constituted are a challenge for the food industry. Thus, the objective of this research was to study the effect of the addition of whey protein concentrate (WPC) in the sensory profile and acceptability of dairy desserts free-fat. Four formulations had been developed with different WPC concentrations (0%, 1.5%, 3.0% or 4.5%, mass%) and sensory characterized by Descriptive Analysis. The acceptability of the formulations was evaluated by 94 consumers who had used the nine-point hedonic scale. Dairy desserts were characterized by eight sensory attributes: color, firmness, resistance to cut, brightness, thickness, gumminess, aroma and vanilla flavor. The formulations containing 3.0 and 4.5% of WPC presented greater intensity of all the attributes. The addition of WPC was effective in the composition of the texture of free-fat dairy desserts and enhances color, flavor and aroma of the product. The formulations with 1.5 and 3.0% WPC had been most accepted, showing that the use of WPC in these concentrations favored the acceptability of the product, providing pleasant sensory characteristics to consumers.