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We compared protein profiles of the synovial fluid of patients with rheumatoid arthritis and osteoarthritis by using surface-enhanced laser desorption/ionization mass spectrometry technology. With this approach, we identified a protein expressed specifically in the synovial fluid of the patients with rheumatoid arthritis. During the investigation, we found several reproducible and discriminatory biomarker candidates for distinction between rheumatoid arthritis and osteoarthritis. Among these candidates, a 10 850 Da protein peak was the clearest example of a single signal found specifically in the rheumatoid arthritis samples. This candidate was purified using a size-exclusion spin column followed by gel electrophoresis and subsequently identified by peptide mapping and post-source decay (PSD) analysis. The results clearly indicate that the protein is myeloid-related protein 8, which was verified by the enzyme immunoassay. It is known that the myeloid-related protein 8 level in serum and synovial fluid is related to disease activity in juvenile rheumatoid arthritis. The results suggest that the ProteinChip platform is useful to detect and identify protein biomarkers expressed specifically in diseases or in some stage of diseases.

Gastric parietal cells migrate from the luminal to the basal region of the gland, and they gradually lose acid secretory activity. So far, distribution and function of K ؉ -Cl ؊ cotransporters (KCCs) in gastric parietal cells have not been reported. We found that KCC3a but not KCC3b mRNA was highly expressed, and KCC3a protein was predominantly expressed in the basolateral membrane of rat gastric parietal cells located in the luminal region of the glands. KCC3a and the Na ؉ ,K ؉ -ATPase ␣1-subunit (␣1NaK) were coimmunoprecipitated, and both of them were highly localized in a lipid raft fraction. The ouabainsensitive K ؉ -dependent ATP-hydrolyzing activity (Na ؉ ,K ؉ - ATPase activity) was significantly inhibited by a KCC inhibitor (R-(؉)-[(2-n-butyl-6,7-dichloro-2-cyclopentyl-2,3-dihydro-1-oxo-1H-inden-5-yl)oxy]acetic acid (DIOA)). The stable exogenous expression of KCC3a in LLC-PK1 cells resulted in associ-ation of KCC3a with endogenous ␣1NaK, and it recruited ␣1NaK in lipid rafts, accompanying increases of Na ؉ ,K ؉ -ATPase activity and ouabain-sensitive Na ؉ transport activity that were suppressed by DIOA, whereas the total expression level of ␣1NaK in the cells was not significantly altered. On the other hand, the expression of KCC4 induced no association with ␣1NaK. In conclusion, KCC3a forms a functional complex with ␣1NaK in the basolateral membrane of luminal parietal cells, and it up-regulates ␣1NaK in lipid rafts, whereas KCC3a is absent in basal parietal cells.

Mice were fed a control diet or a diet supplemented with hyodeoxycholic acid, the most abundant bile acid contained in pig bile, for 4 weeks, after which their serum and livers were collected. The contents of total fatty acids of serum and liver cholesteryl esters, and of liver triglycerides, were reduced following the administration of the hyodeoxycholic acid-supplemented diet, which was mainly due to the reductions in the contents of monounsaturated fatty acids. Free cholesterol contents in the serum and liver were not changed by hyodeoxycholic acid administration. Hyodeoxycholic acid administration reduced the gene expression levels of sterol regulatory element binding protein 1c, acetyl-CoA carboxylase, fatty acid synthase, and stearoyl-CoA desaturase-1. Hyodeoxycholic acid administration markedly changes the ratio of FXR-antagonist/FXR-agonist bile acids in the enterohepatic tissues of the mice (1.13 and 7.60 in hyodeoxycholic acid and control diet groups, respectively). Our findings demonstrate that hyodeoxycholic acid administration exerts the hypolipidemic effect in mice, in which downregulations of de novo lipogenesis and desaturation of saturated fatty acids are suggested to play important roles. In addition, regulation of FXR activation through the selective modification of the enterohepatic bile acid pool may be involved in the hypolipidemic effect of hyodeoxycholic acid administration.

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Highlights d Both the ciliate host and the algal endosymbiont gain fitness benefits d The major benefit for the host is prolonged survival under starvation d The endosymbiont benefits from host's phagotrophy and predation on the competitors d Fitness feedback between the partners evolves cooperation in algal endosymbioses

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