RESUMEN
Oversulfated chondroitin sulfate (OSCS), an impurity found in some porcine intestinal heparin samples was separated from intact heparin by capillary electrophoresis (CE) using a 600mM phosphate buffer, pH 3.5 as the background electrolyte in a 56cm x 25microm i.d. capillary. This method was confirmed in two separate labs, was shown to be linear, reproducible, robust, easy to use and provided the highest resolution and superior limits of detection compared to other available CE methods. Glycosoaminoglycans such as dermatan sulfate and heparan sulfate were separated and quantified as well during a single run. The heparin peak area response correlated well to values obtained using the official assay for biological activity. A high speed, high resolution version of the method was developed using 600mM lithium phosphate, pH 2.8 in a 21.5cm x 25microm i.d. capillary which provided limits of detection for OSCS that were below 0.1%.
Asunto(s)
Anticoagulantes/química , Sulfatos de Condroitina/química , Dermatán Sulfato/química , Contaminación de Medicamentos , Heparina/química , Heparitina Sulfato/química , Fosfatos/química , Animales , Anticoagulantes/análisis , Tampones (Química) , Calibración , Sulfatos de Condroitina/análisis , Sulfatos de Condroitina/aislamiento & purificación , Dermatán Sulfato/análisis , Dermatán Sulfato/aislamiento & purificación , Electrólitos/química , Electroforesis Capilar/instrumentación , Electroforesis Capilar/métodos , Heparina/análisis , Heparitina Sulfato/análisis , Heparitina Sulfato/aislamiento & purificación , Concentración de Iones de Hidrógeno , Estructura Molecular , Concentración Osmolar , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Sus scrofa , Temperatura , Factores de TiempoRESUMEN
Epidemiological evidence suggests a link between chronic oxygen starvation and fat accumulation/obesity, however the underlying mechanism remains unclear. Using Caenorhabditis elegans we found extended oxygen deprivation resulted in activation of SBP-1, the worm homologue of SREBP1, a transcription factor important in maintaining lipid homeostasis. SBP-1 knockdown prevented hypoxia-induced fat accumulation and the associated increase in worm width/length ratio, demonstrating that SBP-1/SREBP1 plays an essential role in hypoxia-induced lipid accumulation and body shape alteration. This study provides the first evidence suggesting that activation of SREBP1 may be a critical pathogenic factor contributing to chronic hypoxia associated excessive fat accumulation/obesity in humans.