RESUMEN

The purpose of the study was to examine the potential of inhibition of cathepsin S as a treatment for autoimmune diseases. A highly selective cathepsin S inhibitor, CSI-75, was shown to upregulate levels of the cathepsin S substrate, invariant chain Lip10, in vitro as well as in vivo in C57Bl/6 mice after oral administration. Functional activity of the compound was shown by a reduction in the OVA-specific response of OVA-sensitized splenocytes from C57Bl/6 mice as well as from OVA-TCR transgenic mice (DO11.10). Since these studies revealed a selective suppression of the Th1 and Th17 cytokines causing a shift to Th2, CSI-75 was tested in the murine HC-gp39-immunization model. Indeed, CSI-75 specifically reduced the circulating HC-gp39-specific IgG2a in these mice indicating selective inhibition of the Th1 type of response in vivo. The importance of especially the Th1 and Th17 cell subsets in the pathology of autoimmune diseases, renders CatS inhibition a highly interesting potential therapeutic treatment of autoimmune diseases. Therefore, CSI-75 was tested in a murine model of multiple sclerosis (i.e. experimental autoimmune encephalomyelitis (EAE)) in a semi-therapeutic setting (ie. oral treatment after initial sensitization to antigen). Finally, in a murine model with features resembling rheumatoid arthritis (the collagen-induced arthritis (CIA) model), CSI-75 was tested in a therapeutic manner (after disease development). CSI-75 caused a significant reduction in disease score in both disease models, indicating a promising role for CatS inhibitors in the area of therapeutic treatments for autoimmune diseases.

Asunto(s)

Enfermedades Autoinmunes/tratamiento farmacológico , Catepsinas/antagonistas & inhibidores , Piperidinas/uso terapéutico , Inhibidores de Proteasas/uso terapéutico , Piridinas/uso terapéutico , Administración Oral , Animales , Células Presentadoras de Antígenos/efectos de los fármacos , Artritis Experimental/tratamiento farmacológico , Enfermedades Autoinmunes/inmunología , Encefalomielitis Autoinmune Experimental/tratamiento farmacológico , Femenino , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Piperidinas/administración & dosificación , Inhibidores de Proteasas/administración & dosificación , Piridinas/administración & dosificación , Células TH1/fisiología

RESUMEN

Past publications have highlighted the influence of postdialysis plasma pH on the measured fraction unbound in plasma (fup). There is disparity in the industry as to which of two main methods is more suitable for controlling postdialysis plasma pH: the use of either a stronger buffer or a CO(2) atmosphere for the incubation. In the current study, it has been found that 10% CO(2) could be too high for the buffering capacities of both 100 mM sodium phosphate (pH 7.40 decreased to pH 6.90 after a 6-h incubation) and plasma (decreased below pH 7.40 after a 6-h incubation). To provide appropriate control over the postdialysis plasma pH, for a range of species, it is proposed that a standard phosphate buffer strength (100 mM) and pH (7.40) in combination with a 5% CO(2) atmosphere be used for equilibrium dialysis. Furthermore, statistically significant differences in fup values obtained with a pH difference of less than 0.32 pH unit have been demonstrated. An acceptance range for postdialysis plasma pH in routine in vitro fup screening assays of pH 7.40 ± 0.10 is recommended.

Asunto(s)

Proteínas Sanguíneas/metabolismo , Preparaciones Farmacéuticas/sangre , Animales , Tampones (Química) , Dióxido de Carbono , Fenómenos Químicos , Diálisis , Perros , Evaluación Preclínica de Medicamentos/métodos , Humanos , Concentración de Iones de Hidrógeno , Macaca fascicularis , Ratones , Concentración Osmolar , Preparaciones Farmacéuticas/química , Preparaciones Farmacéuticas/metabolismo , Unión Proteica , Ratas , Reproducibilidad de los Resultados

RESUMEN

The averaged point-spread function (PSF) estimation of an image acquisition system is important for many computer vision applications, including edge detection and depth from defocus. The paper compares several mathematical models of the PSF and presents an improved measurement technique that enables subpixel estimation of 2D functions. New methods for noise suppression and uneven illumination modeling were incorporated. The PSF was computed from an ensemble of edge-spread function measurements. The generalized Gaussian was shown to be an 8 times better fit to the estimated PSF than the Gaussian and a 14 times better fit than the pillbox model.