Regular structures on the microvillar surface membrane of ileal epithelial cells in suckling rat intestine

Freeze-fracture deep-etch studies were done to examine regular patterns of surface membrane particles in the suckling rat ileal epithelium by using quick-freezing method. In addition to the presence on the luminal surfaces of well-developed endocytic complexes, latticed particles were consistently observed on almost the entire apical membrane between microvilli and frequently on the microvilli. These particles seemed to be partly integrated in the outer half layer of membranes. After rinsing with phosphate-buffered saline, patterns on the microvilli appeared to be parallel circle lines rather than latticed particles found in fresh preparations. Filipin treatment showed the presence of filipin-sterol complexes on most of the particle-covered membrane areas except small tubules and vesicles. Possibilities were suggested that these latticed particles were transferred to the apical surface along the outer half layer of membranes, and released or secreted into the intestinal lumen.     

Inflammation-driven senescence-associated secretory phenotype in cancer-associated fibroblasts enhances peritoneal dissemination

1. Download : Download high-res image (264KB)
2. Download : Download full-size image

     

Determination of collagen band positions on polyacrylamide gel without staining

Protein band positions on a slab polyacrylamide gel were determined without staining, by blotting the bands onto a nitrocellulose membrane which was then stained to visualize the electrophoretic pattern. The unfixed, unstained bands were cut from the gel according to the copy pattern placed underneath the slab gel. Collagen alpha 1 and alpha 2 chains were separately extracted from the gel using this procedure.     

Calcitonin inhibits the growth of human gastric carcinoma cell line KATO III.

Calcitonin has a wide variety of actions on gastrointestinal function. In this study, we investigated the effects of calcitonin on the growth of human gastric carcinoma cell line KATO III in comparison with those of calcitonin gene-related peptide (CGRP). Calcitonin, but not CGRP, significantly and dose-dependently inhibited the growth of KATO III cells. This inhibition of cell growth was accompanied by an increase in cyclic AMP production. The proliferation of KATO III cells was also inhibited by forskolin and dibutyryl cyclic AMP, although agents which do not stimulate cyclic AMP production had no effect. Furthermore, in the presence of GTP, calcitonin stimulated adenylate cyclase activity in KATO III cell membranes, and this increase was reduced in the absence of GTP. On the other had, neither calcitonin nor CGRP enhanced the turnover of inositolphospholipid or the intracellular Ca2+ level. In addition, 125I-labeled human calcitonin was specifically bound to KATO III cell membranes, and this binding was dose-dependently displaced by unlabeled calcitonin but not CGRP. Furthermore, the specific binding of 125I-labeled human calcitonin to KATO III cell membranes was significantly reduced by addition of GTP but not ATP. These results suggest that calcitonin inhibits the growth of human gastric carcinoma cell line KATO III by stimulating cyclic AMP production via a GTP-dependent process coupled to specific calcitonin receptors.     

Isolation and characterization of 3-N-trimethylamino-1-propanol-degrading Rhodococcus sp. strain A2

The aerobic degradation of 3- N -trimethylamino-1-propanol (homocholine) as a sole source of carbon and nitrogen has been found for a Rhodococcus sp. bacterium isolated from soil. The isolate was identified as Rhodococcus sp. strain A2 based on its phenotypic features, physiological and biochemical characteristics, and results of phylogenetic analysis. The washed cells of strain A2 completely degraded homocholine within 6 h, with concomitant formation of several metabolites. Analysis of the metabolites using capillary electrophoresis, fast atom bombardment–MS, and GC–MS showed that trimethylamine was the major metabolite, in addition to β-alanine betaine (β-AB) and trimethylaminopropionaldehyde. Therefore, the possible degradation pathway of homocholine in the isolated strain is through consequent oxidation of the alcohol group (-OH) to aldehyde (-CHO) and acid (-COOH). Thereafter, the cleavage of β-AB C–N bonds yielded trimethylamine and alkyl chain.