An application of the 2-nitrobenzenesulfenyl method to proteomic profiling of human colorectal carcinoma: A novel approach for biomarker discovery

In the development of novel biomarkers, the proteomic approach is advantageous because using it the cancer-associated proteins can be directly identified. We previously developed a 2-nitrobenzenesulfenyl (NBS) method to improve quantitative proteome analysis. Here, we applied this method to proteomic profiling of colorectal carcinoma (CRC) to identify novel proteins with altered expression in CRC. Each pair of tumor and normal tissue specimens from 12 CRC patients was analyzed, and approximately 5000 NBS-labeled paired peaks were quantified. Peaks with altered signal intensities (>1.5-fold) and occurring frequently in the samples (>70%) were selected, and 128 proteins were identified by MS/MS analyses as differentially expressed proteins in CRC tissues. Many proteins were newly revealed to be CRC related; 30 were reported in earlier studies of CRC. Six proteins that were up-regulated in CRC (ZYX, RAN, RCN1, AHCY, LGALS1, and VIM) were further characterized and validated by Western blot and immunohistochemistry. All six were found to be CRC-localized, either in cancer cells or in stroma cells near the cancer cells. These results indicate that the proteins identified in this study are novel candidates for CRC markers, and that the NBS method is useful in proteome mining to discover novel biomarkers.     

Growth and development of colorectal cancer with subclassification on submucosa and proper muscle infiltrating cancer

Colorectal cancer (CRC) infiltrating the submucosa (sm cancer) and the proper muscle (mp cancer) represent an early and an intermediate stage in the development of CRC, and sm and mp cancer are therefore appropriate to discuss the natural history of CRC. A total of 337 sm cancers and 291 mp cancers resected in our series were evaluated. We divided sm and mp cancers into three categories, respectively, sm 1, sm 2, sm 3, mp 1, mp 2, and mp 3, according to the depth of infiltration of the submucosa or the proper muscle. The deeper the invasion, the larger of the tumor size, and the proportion of depressed type in their configuration increases. On the contrary, the proportion of tumors with adenomatous component decreases. A larger proportion of tumors in all categories of infiltration showed PG more of ten than that of NPG in their marginal structure. In this study, 17% of sm cancer and 23% of mp cancer might develop not via the adenoma-carcinoma sequence.     

Immunofluorescence localization of a 25-kDa Tetrahymena EF-hand Ca(2+)-binding protein, TCBP-25, in the cell cortex and possible involvement in conjugation

The Tetrahymena Ca(2+)-binding protein of 25 kDa (TCBP-25) is a member of the calmodulin family containing four EF-hand Ca(2+)-binding loops, but its biological role has not yet been investigated. In this study, TCBP-25 was expressed in Escherichia coli as a glutathione S-transferase fusion protein and then purified. Purified TCBP-25 showed a typical Ca(2+)-dependent shift in electrophoretic mobility, consistent with conformational change caused by Ca(2+)-binding. Localization of TCBP-25 was examined by indirect immunofluorescence using an antiserum specific for TCBP-25. Strong immunofluorescence was observed all over the cell cortex except in and around basal bodies. From the results of immunofluorescence using detergent-extracted cells, TCBP-25 is suggested to exist as an insoluble form in the cell cortex. TCBP-25 appears to be localized in the cortical alveoli or the epiplasm and exists around both the migratory and the stationary gametic pronuclei at the pronuclear exchange stage during conjugation. Therefore, we speculate that TCBP-25 may play crucial roles in Ca(2+)-mediated signaling processes in the cell cortex and in a Ca(2+)-dependent pronuclear exchange process during conjugation.     

Effect of shear thinning on aspiration – Toward making solutions for judging the risk of aspiration

Polysaccharide solutions containing a contrast medium were prepared so that their viscosity at 50 s⁻¹, which was reported as a shear rate in the oral cavity, is the same but different at other shear rates. Preparation procedure of such solutions based on heat treatment using widely used polysaccharides in foods is described, and the rheological properties of these solutions are discussed in relation with flow behaviours in the swallowing process. Videofluorographic observation was performed for healthy persons and patients and compared with rheological characteristics of these solutions to understand the mechanism of aspiration. It was shown that the degree of aspiration depended on the degree of shear thinning of polysaccharide solutions. This study showed the possibility to find what kind of fluid induces the aspiration and to reduce the risk of aspiration in dysphagic patients by controlling the food rheology.     

金属推块V-带型CVT的研究Ⅱ——金属推块和环拉伸之间的压缩力

本研究采用超小型测力仪插入两推块和遥测装置之间的在几种固定速比下测量主动劝带轮上推块压力分布。同时采用一种特别 推则出了环拉力的分布。从实验结果可以作出以下结论。     

金属V-带推块型CVT的研究(Ⅰ)——传递转矩和带轮推力间关系

本文在金属Ｖ－带推块型ＣＶＴ上,测出了主、从动带轮的传递转矩和推力以及峡谷带轮间的轴向力。绘出了不同速比下主,从动带轮间相对于转矩比的比线图并作相互比较。发现推力比与速比几乎和带轮转速及正常转矩下的最大可传递的转矩无关,该推力比是速比的函数,它和转矩比及推块与带轮间的摩擦系数有关,还导出了带轮推力经验平衡方程式,该式以显函数形式表示。它可以简化并可适用于电控ＣＶＴ。     

Effects of inner-phase components of water-in-oil-in-water emulsion on low-pH tolerance of Lactobacillus acidophilus incorporated into inner-water phase

The incorporation of Lactobacillus acidophilus, which has a low acid tolerance, into the inner-water phase of water-in-oil-in-water (W/O/W) emulsion improved the bacterial survival rate in a model gastric juice. The components that enhanced the acid tolerance of the bacterium in W/O/W emulsion were investigated. Although the acid tolerance enhancement induced by the tested components was low at pH 2, some components significantly improved the tolerance at pH 4. In particular, the acid tolerance enhancement induced by yeast extract and sodium caseinate was marked.     

Formation equation of halogenated organic compounds when water is chlorinated

Various halogenated organic compounds are formed by chlorination of water. In this study, formation of organic compounds halogenated from a reagent humic acid and extract of a leaf mold were examined under various conditions. The following overall formation equation was obtained from empirical data under the practical wide range when free chlorine remained.

[TOX]=k_TOX[TOC][Cl₂]_ot^β.

Here, [TOX] is the concentration of total organic halogen after t h in units of mg chlorine per liter; [TOC] and [Cl₂]_o are concentrations of total organic carbon and dosed chlorine in units of mg per liter; k_TOX is the rate constant and and β are parameters. From the values of k_TOX, and β, the character of organic substances i.e. precursor of halogenated organic compounds, in water can be evaluated. The values k_TOX, and β for humic acid are 0.053, 0.28 and 0.13, and the values for extract of the leaf mold are 0.032, 0.36 and 0.15, respectively. The activation energies are 10 kJ mol⁻¹ and 11 kJ mol⁻¹ for the reactions of humic acid and leaf mold extract, respectively.