Our investigation is one of the first to critically analyze the factors in presentations at annual meetings that predict successful publication.
The acute phase response to inflammation induces changes in the secretion of hepatic proteins. To examine the time course of an acute phase protein response in broiler chickens, the plasma levels of hemopexin (HX) and alpha1-acid glycoprotein (AGP) and liver HX mRNA were measured at various time points from 3 hr to 336 hr after an intraabdominal injection of either lipopolysaccharide (LPS), complete Freund's adjuvant, incomplete Freund's adjuvant, phytohemagglutin, or mineral oil. Uninjected chicks served as controls. The accumulation of liver HX mRNA began within 3 hr of stimulation and peaked at 12 hr. Relative to control levels, plasma HX and AGP levels increased by 6-12 hr postchallenge and peaked at 24 hr. Complete Freund's adjuvant and LPS treatments induced the greatest increase in plasma HX (threefold; P < 0.05). Plasma levels of HX and AGP returned to control levels at 336 and 168 hr postinjection, respectively. A second experiment demonstrated that turpentine induced a similar AGP response as LPS and that albumin is a negative acute phase protein. The results suggest that plasma levels of HX or AGP could be used as an indicator of the systemic component of a local inflammatory response in chickens.
Western blots, enzyme assays, protein glycosylation studies, and immunohistochemical staining were used to characterize cathepsin B expression at successive stages of colorectal tumor progression. In normal colon mucosa and premalignant adenomas, cathepsin B expression was predominantly due to mature two-chain protein detected on Western blots as the nonglycosylated 27-kDa form, with overexpression of this protein occurring in only 4 of 18 adenomas. Overexpression increased significantly in Dukes A and B carcinomas (26 of 37 cases), with cathepsin B protein generally detectable in carcinomas as a combination of both 27-kDa nonglycosylated and 28-kDa glycosylated mature two-chain forms. Glycosylated cathepsin B protein in carcinoma extracts was sensitive to PNGase F but resistant to Endo H, indicating a pattern consistent with complex rather than high mannose type glycosylation. When sorted by advancing tumor stage, peak expression of cathepsin B protein occurred in carcinomas involved in local invasion compared with adenomas or metastatic cancers. At all stages, cathepsin B activity correlated significantly with the levels of heavy chain mature cathepsin B protein (r ؍ 0.6682, p < 0.0001) irrespective of glycosylation. Immunohistochemical staining of cathepsin B protein revealed fine diffuse cytoplasmic staining in both adenomas and carcinomas compared with coarse granular cytoplasmic staining (typical of lysosomes) seen in matched normal mucosa. Our results demonstrate several sequential, apparently independent changes in cathepsin B expression during colorectal tumor progression including early changes in subcellular localization, up-regulation of cathepsin B protein and activity in invasive cancers, and altered protein glycosylation detected in malignant tumors at all stages.Among lysosomal cysteine proteinases, increased or altered cathepsin B expression has been particularly well documented in a variety of tumor cell types including colon, breast, pancreas, lung, and brain tumors (1-9). Cathepsin B has also been shown to be relocated to the plasma membrane (9) or secreted from tumor cells (10, 11), where it is believed to aid in degradation of components of the extracellular matrix and basement membrane (12). Furthermore, tumor cathepsin B has also been shown to retain greater proteolytic activity at or above neutral pH (1, 12) than the normal enzyme.Previous studies of cathepsin B expression using a primary tumor model of colorectal cancer progression found cathepsin B enzyme activity to be frequently and significantly elevated in the early invasive stages of tumor growth (1, 2, 4). This finding was confirmed by Northern analyses of cathepsin B mRNA in colorectal tumors that also demonstrated greater increases in earlier than later stage carcinomas (3). Early elevation in cathepsin B expression, followed by an inverse correlation with cancer progression, suggested that cathepsin B activity might be particularly important in cancers associated with invasion of the colonic wall. These findings have been su...
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