Identification of antigenic epitopes recognized by Mac-2 binding protein-specific cytotoxic T lymphocytes for use in cancer immunotherapy

We have previously reported that 90K/Mac-2 binding protein (M2BP) was highly expressed in lung cancer and that M2BP-specific immunity was observed in many of cancer patients. In this study, we analyzed the ability of 11 M2BP-derived oligopeptides with an HLA-A*0201-binding motif to induce M2BP-specific cytotoxic T lymphocytes (CTL) from peripheral blood lymphocytes of normal donors by in vitro stimulation. One of the CTLs that were induced using M2BP216-224 (RIDITLSSV) produced interferon-gamma in response to HLA-A2-positive T2 cells pulsed with the same peptide and lysed MDA-MB-231 cells expressing both M2BP and HLA-A2. The cytolytic activities were blocked by antibodies against HLA class I or CD8. These findings suggest that M2BP216-224 is naturally processed from the native M2BP in cancer cells and recognized by M2BP-specific CTLs in an HLA-A2 restriction. We first identified M2BP-derived CTL epitopes that may be useful as a target antigenic epitope in clinical immunotherapy of cancer.     

Structural and functional analysis of a lycopene beta-monocyclase gene isolated from a unique marine bacterium that produces myxol

A gene coding for lycopene beta-monocyclase, which metabolizes lycopene (psi,psi-carotene) to gamma-carotene (beta,psi-carotene), was isolated for the first time from a unique marine bacterium strain P99-3 that produces myxol (a gamma-carotene derivative). This lycopene beta-monocyclase gene (designated crtYm) was included in the gene cluster which contained carotenoid biosynthetic gene (crtI, crtB, crtZ, crtY, and crtA) homologs. CrtYm, the CrtY homolog, metabolized lycopene to gamma-carotene, which was confirmed by deletion/expression analysis of the crtYm and by subsequent analysis of the metabolites from lycopene based on the retention times on high-performance liquid chromatography, UV-visible absorption spectra, and mass spectrometry.     

Spatial heterogeneity of TNF-alpha-induced T cell migration to colonic mucosa is mediated by MAdCAM-1 and VCAM-1

Relatively little is known about how recirculation of lymphocytes through the inflamed intestinal mucosa is regulated. The aim of this study was to investigate the dynamic process of T lymphocyte-endothelial cell adhesion in TNF-alpha-challenged murine colonic mucosa by intravital microscopy. T lymphocytes from spleen (SPL) and intestinal lamina propria (LPL) were fluorescence labeled, and their adhesion to microvessels in the colonic mucosa was observed. In TNF-alpha (25 microg/kg)-stimulated colonic venules, an enhanced adhesion of SPL and LPL was demonstrated, with dominant recruitment of LPLs. The magnitude of the increased LPL adhesion was more significant in the colon than in the small intestine. These T lymphocyte interactions in the colonic mucosa were significantly reduced by blocking MAbs against either mucosal addressin cell adhesion molecule-1 (MAdCAM-1), VCAM-1, alpha(4)-integrin, or beta(7)-integrin but not by anti-ICAM-1. Immunohistochemistry revealed significant MAdCAM-1 expression in the lamina propria and VCAM-1 expression in the submucosa of TNF-alpha-treated colon. Spatial heterogeneity of MAdCAM-1 and VCAM-1 activation following TNF-alpha challenge may promote specific T lymphocyte recruitment in the inflamed colonic mucosa.     

Cultivation of microalgae in the solution from the desalting process of soy sauce waste treatment and utilization of the algal biomass for ethanol fermentation

The growth of four marine microalgae was examined in a medium prepared from the effluent from the desalting process of soy sauce waste. A strain of Dunaliella that showed abundant growth on soy sauce waste extract was selected, and optimum cultivation conditions were determined. The algal cells produced were disrupted, and saccharified with glucoamylase. The saccharified solution was fermented using Saccharomyces cerevisiae IAM 4140. Stoichiometric study revealed that 11mg of ethanol was produced from 1g (dry cell weight) of Dunaliella cells. This work indicates a new method for removing waste products of the soy sauce production industry.     

Random disturbance and diversity of competitive systems

Population size distribution within a community of species with a competitive hierarchy, is studied. With the introduction of a stochastic model of invasion and extinction due to random environmental disturbance, the stationary population size distribution is derived as a function of the parameter, which denotes the ratio of rates of random extinction and invasion. From this result it is shown that the Shannon-type diversity takes the maximum value under the situation that 10 20 percent of habitat sites remain unoccupied, and the number of species also becomes maximum at intermediate levels of disturbance. The dynamical properties of a one-sided competitive system are also discussed by use of the Lotka-Volterra equations.     

Ultrastructural distribution of growth hormone and prolactin mRNAs in normal rat pituitary cells: a comparison between preembedding and postembedding methods

In situ hybridization (ISH) at the electron microscopic level is essential for elucidating the intracellular distribution and role of mRNA in protein synthesis. We describe our electron microscopic ISH method using biotinylated oligonucleotide probes for rat growth hormone and prolactin mRNAs and compare the preembedding method with the postembedding method. Preembedding electron microscopic ISH localized rat growth hormone and prolactin mRNAs on the polysomes of the rough endoplasmic reticulum (RER). Rat growth hormone mRNA was distributed diffusely on the RER, whereas rat prolactin mRNA was scattered and distributed focally. Thus there might be a specific translational site for prolactin mRNA on the RER. Rat growth hormone mRNA signals were also recognized on the polysomes of the RER, using the postembedding method with streptavidin gold conjugate. The hybridization signal intensity using the postembedding method was lower, and non-specific signals were more frequent, in comparison with the preembedding method. The preembedding method thus appears to be easier and better than the postembedding method from the viewpoint of utility and preservation of mRNA. Electron microscopic ISH is considered to be an important tool for evaluating the intracellular localization of mRNA and the site of specific hormone synthesis on the RER.     

Increasing glutaminase activity in shoyu koji using a mixed culture of two koji moulds

For improved fermentation of shoyu (soy sauce), a useful koji-making system has been developed using a mixed tane-koji of two shoyu koji moulds, namely Aspergillus oryzae K2 (length of conidiophores about 350 m) and the late-conidiation strain, A. oryzae HG (length of conidiophores about 2500 m). The mixed culture of strains K2 and HG had about twice the glutaminase activity of the single-strain cultures. In addition, the number of conidia in the mixed culture was about 10% of that in a culture of strain K2 alone.     

Facilitative effect of a novel AVP fragment analog, NC-1900, on memory retention and recall in mice

In order to determine the mechanism of action of a new AVP(4-9) analog, NC-1900, on memory processes, memory retention and retrieval tests were conducted in a step-through passive avoidance (PA) task in mice. The administration of NC-1900 facilitated memory retention and retrieval in the PA task through vasopressin1A (V1A) receptors but not V2 receptors. The effect of NC-1900 on memory retention test performance appeared to be due to activation of the protein kinase C (PKC) signaling pathway via V1A receptors; however, the modulation of PKC was not essential for the facilitative effect of the new peptide in the retrieval test. The facilitation of memory retrieval by NC-1900 may also be mediated by other non-PKC-dependent signaling pathways, such as the phospholipase C-inositol trisphosphate pathway.     

Advantage of dichromats over trichromats in discrimination of color-camouflaged stimuli in nonhuman primates

Due to a middle- to long-wavelength-sensitive (M/LWS) cone opsin polymorphism, there is considerable phenotypic variation in the color vision of New World monkeys. Many females have trichromatic vision, whereas some females and all males have dichromatic vision. The selective pressures that maintain this polymorphism are unclear. In the present study we compared the performance of dichromats and trichromats in a discrimination task. We examined tri- and dichromatic individuals of two species: brown capuchin monkeys (Cebus apella) and long-tailed macaques (Macaca fascicularis). We also examined one protanomalous chimpanzee (Pan troglodytes). The subjects' task was to discriminate a circular pattern from other patterns in which textural elements differed in orientation and thickness from the background. After they were trained with stimuli of a single color, the subjects were presented with color-camouflaged stimuli with a green/red mosaic overlaid onto the pattern. The dichromatic monkeys and the protanomalous chimpanzee selected the correct stimulus under camouflaged conditions at rates significantly above chance levels, while the trichromats did not. These findings demonstrate that dichromatic nonhuman primates possess a superior visual ability to discriminate color-camouflaged stimuli, and that such an ability may confer selective advantages with respect to the detection of cryptic foods and/or predators.