The cooperative induction of macrophage activation by fucoidan derived from Cladosiphon okamuranus and ß-glucan derived from Saccharomyces cerevisiae.

The present study investigated immune stimulatory effects of Cladosiphon okamuranus-derived fucoidan to activate murine macrophage-like cell line RAW264, and the functional relationship with zymosan, a Saccharomyces cerevisiae-derived ß-glucan. The production of nitric oxide (NO) and tumor necrosis factor-α (TNF-α) in RAW264 cells were remarkably enhanced in the presence of 10 µg/mL fucoidan, and the stimulatory effects of fucoidan were maximally augmented in combinational treatment with 500 ng/mL zymosan, whereas any TLR ligands had no those effects. Confocal microscopic analyses suggested that fucoidan bound on plasma membrane, and it was estimated that some cell surface molecules acted as receptor for fucoidan because cytochalasin D, an inhibitor of phagocytosis, did not affect the immune enhancing activities, whereas methyl-ß-cyclodextrin (MßCD), a general agent for disruption of lipid rafts, diminished that. Furthermore, it was revealed that the additive effects of zymosan on the immune activation with fucoidan was thought to be mediated by dectin-1 based on the results with dectin-1-knockdown RAW264 cells. All of results suggested that fucoidan and some kinds of ß-glucan would cooperatively reinforce the activity of innate immune cells via interactive receptor crosstalk.

Ex vivo induction of mRNA in human whole blood as a new platform of drug and dietary supplement development.

PURPOSE: We introduced a new concept of ex vivo gene expression analysis (Mitsuhashi, Clin Chem 53:148-149, 2007), where drug action was simulated under physiological conditions. This model system was applied to study various fields of drug development. MATERIALS AND METHODS: Heparinized human whole blood was incubated with drugs for less than 4h. The changes of specific mRNA were then quantified using the method we developed (Mitsuhashi, Tomozawa, Endo, and Shinagawa, Clin Chem 52:634-642, 2006). RESULTS: The mRNA quantitation method was used as a model system to study the following areas: (1) identification of respondents and non-respondents, (2) ex vivo compound screening, (3) determination of individually optimized doses, (4) drug-to-drug comparison, (5) assessment of leukocyte toxicity, (6) discovery of molecular targets, (7) assessment of the action of dietary supplements, and (8) characterization of respondents and non-respondents for various dietary supplements. CONCLUSION: Since ex vivo assays are safe, a large number of healthy donors and disease patients can be recruited to identify individual-to-individual variations, which is not available from current preclinical study models. Although each system should be validated using a large number of samples, the ex vivo analysis will be a new tool for the development of drugs and dietary supplements in future.

A clinicopathological investigation of "tumor nodules" in colorectal cancer.

PURPOSE: Tumor nodules (tn) have been histologically identified within the fatty tissue or the detached fatty tissue around dissected lymph nodes, or else picked up as lymph nodes from resected specimens with no lymph node components. The TNM classification of malignant tumors provides a description of how to deal with tn, but there has so far been no description within the Japanese classification of colorectal carcinoma. The aim of this study was to determine whether we should regard tn as metastatic lymph nodes from the viewpoint of prognosis. METHODS: A total of 544 patients who underwent a resection of colorectal adenocarcinoma between 1985 and 1995 were reviewed. RESULTS: Tumor nodules were found in 54 (17.6%) of 307 colon cancer patients, and in 41 (17.3%) of 237 rectal cancer patients. We classified the curability A patients into four groups for both colon and rectal cancer; positive lymph nodes with tn (Group A), negative lymph nodes with tn (Group B), positive lymph nodes without tn (Group C), and negative lymph nodes without tn (Group D). The prognosis was not significantly different between Groups A, B, and C, but it was significantly different between Group D and Groups A, B, and C (P < 0.01) in both the colon and the rectum. CONCLUSION: From the viewpoint of prognosis, it thus appears justifiable to regard tn as lymph node metastasis.