NR AVFD
AU Zhou,X.B.; Gao,J.M.; Gao,C.; Zhang,B.Y.; Yuan,Y.K.; Dong,X.P.
TI [Influence of ultrasonic processing on the aggregation of PrP-Sc in the brain extracts of the scrapie-infected hamsters]
QU Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese Journal of Experimental and Clinical Virology 2004 Jun; 18(2): 118-21
PT journal article
AB BACKGROUND: To evaluate the influence ultrasonic processing on the aggregation of PrPsc in the brain extracts prepared from the scrapie-infected hamsters, and to seek for the way to prepare lower molecular PrPsc polymer. METHODS: The extracts of infected brains were prepared with a lysis solution, and treated with ultrasonics at various conditions during the different phases. The distribution and aggregation state of PrPsc were analyzed by proteinase K treated. Western blot, and afterwards, quantitatively calculated with a commercially supplied software Image Totaltech. RESULTS: The amount of PrPsc in the supernatant of brain homogenates was 1.29-to 1.58-fold increased with appropriate sonication (15 s for 30 times). At the same conditions of ultrasound, the PrP amount in the supernatant prepared from the scrapie-infected hamster brain was significantly increased, whereas that prepared from healthy animal used as normal control showed little change. Comparative analyses of PrPsc pellets prepared by high-speed centrifugation revealed that about 90% PrPsc released into supernatant after ultrasound processing. CONCLUSION: Appropriate sonication of homogenate of scrapie-infected brain increases the extracted amount of PrPsc, being favorable to laboratory diagnosis. Larger molecular PrPsc aggregates can be crashed by ultrasonic processing, engendering lower molecular PrPsc polymers.
MH Animals; Blotting, Western; *Brain Chemistry; Cricetinae; English Abstract; Image Processing, Computer-Assisted; PrPsc Proteins/*analysis; Research Support, Non-U.S. Gov't; Scrapie/*metabolism; *Sonication
AD Institute for Viral Disease Control and Prevention, China CDC, Beijing 100052, China.
SP chinesisch
PO China