De-phosphorylation of GR at Ser203 in nuclei associates with GR nuclear translocation and GLUT5 gene expression in Caco-2 cells

Glucocorticoid hormones and p44/42 mitogen-activated protein kinase (MAPK) inactivation are considered to be important in small-intestinal differentiation/maturation. In this study, we found that co-treatment with glucocorticoid hormone agonist dexamethasone and p44/42 MAPK inhibitor PD98059 in intestinal cell line Caco-2 strongly induced GLUT5 gene expression. Glucocorticoid hormone receptor (GR) was translocated from the cytoplasm to the nucleus and de-phosphorylated at serine residue 203 in the nucleus, by combined treatment with dexamethasone and PD98059. The binding of GR, as well as acetylated histones H3 and H4, to the promoter/enhancer region of GLUT5 gene was enhanced by combined treatment with dexamethasone and PD98059. These results suggest that the inactivation of p44/42 MAP kinase enhances glucocorticoid hormone-induced GLUT5 gene expression, probably through controlling the phosphorylation at serine 203 and nuclear transport of GR, as well as histone acetylation on the promoter/enhancer region of GLUT5 gene.     

Dietary resistant starch reduces levels of glucose-dependent insulinotropic polypeptide mRNA along the jejunum-ileum in both normal and type 2 diabetic rats

It has been reported that the circulating glucose-dependent insulinotropic polypeptide (GIP) levels were reduced by an intake of some foods/drugs capable of delaying carbohydrate digestion/absorption. In this study, we revealed that feeding rats with dietary resistant starch reduced the GIP mRNA levels along the entire length of the jejunoileum in both Wistar and type 2 diabetic GK rats.     

A study of polyploidy in races of Japanese weevils (Coleoptera: Curculionidae)

The chromosomes of two Japanese weevils, Blosyrus japonicus Sharp and Callirhopalus (= Pseudocneorhinus) obesus Roelofs, were studied. The former consists of four races: one 2n and bisexual (= Blosyrus falcatus Faust) and three parthenogenetic-4n, 5n, and 6n. In general the races are allopatric. The latter species has two races: 4n and 6n, again with distinct geographical distributions.     

Studies on the Behaviors of Impurities on the Crystallization of l-Glutamic Acid

The behaviors of impurities on the crystallizations of the free acid and the hydrochloride, monosodium, zinc, calcium and barium salts of l-glutamic acid were examined, and a tendency was recognized that coexisting impurities were apt to be taken into the crystals when the crystallization proceeded from the zwitterion, i.e., Glu.^± or The adsorption of l-tyro-sine was compared when l, d and dl-glutamic acid were crystallized with coexistence of l-tyrosine, and an effect of more or less extent of steric configuration on its behavior could be recognized.     

Studies on the Polymorphism of l-Glutamic Acid

The enthalpy differences between the α- and β-crystals of l-glutamic acid were measured and ΔH_α?β of ?75, ?95, 8 and 27±20cal./mol. were obtained at temperatures of 5, 15, 30 and 40°C, respectively. From this result, it was found that the α-form is the more stable form in solid phase at temperatures lower than 30°C and vice versa when higher than 30°C, though the α-form had been found to be the less stable form as the saturating body in the aqueous solution in all range of temperature.     

A New Crystalline form of Monosodium l-Glutamate

A new crystalline form of monosodium l-glutamate, which is different from the known monosodium l-glutamate monohydrate crystal in external appearance, density, X-ray diffraction pattern, solubility, and water of crystallization, has been crystallized from ca. 73 weight per cent aqueous methanol solution at ca. 18°C. It was identified as the dihydrate from the determination of water of crystallization based on Karl Fischer′s method. The newly found crystals are extremely unstable in air, aqueous solution, and the mother liquor, because of rapid transformation into the stable monohydrate, whereas they can be kept in a considerably stable condition in absolute ethanol and acetone.     

Alkaline Earth Salts of Glutamic Acid and Optical Resolution of their Racemic Modifications

The solubilities of monocalcium and monomagnesium diglutamate and calcium, magnesium, barium and strontium glutamate in water were measured in respect to both the l-form and the dl-form to conjecture the constitution of the racemic modification of each salt. It was concluded from the result that only magnesium dl-glutamate among six saltsis a congromerate and the others except the barium salt are all racemic compounds. The conclusion was supported by results from measurements of the infrared spectra and the X-ray diffractions. As for barium glutamate, a definite conclusion could not be obtained, but its racemic form seems to be congromerate or a racemic compound with a weak affinity between the isomers. Optical resolution based on the inoculation method was tried for calcium, magnesium, barium and strontium dl-glutamate, and the selective crystallization of the active form from the supersaturated solutions of magnesium and calcium dl-glutamate proceeded, though the process was unstable in the latter case; on the other hand, the resolutions of barium and strontium dl-glutamates were quite unsuccessful.     

Establishment and characterization of SV40 large T antigen-immortalized cell lines derived from fetal bovine brain tissues after prolonged cryopreservation

Bovine brain cell lines with specific characteristics are useful in vitro experimental systems for molecular and cellular investigation of the interactions between bovine specific neuropathogenic agents and the host. Here, we established two novel cell lines from cultures of cryopreserved fetal bovine brain tissue by the transfection of SV40 large T antigen. Both cell lines showed cobblestone morphology in DMEM/F12 medium supplemented with 10% fetal bovine serum, epidermal growth factor and basic fibroblast growth factor. They were immunostained with endothelial marker, Von Willebrand Factor. Endothelial properties, such as capillary-like tube formation on matrigel and the incorporation of DiI-AcLDL were confirmed with these cells. Removal of growth factors increased the number of cells expressing alpha-smooth muscle actin, suggesting the potential of these cell lines to differentiate into smooth muscle cells. This study suggests an efficient protocol to immortalize brain endothelial cell lines from fetal bovine brain tissue culture.