Influence of corticotropin-releasing factor on pancreatic and islet blood flow in different regions of the rat pancreas

Corticotropin-releasing factor (CRF) has previously been shown to selectively dilate the mesenteric vascular bed, without affecting other vascular beds. Pancreatic blood flow and islet blood flow were therefore measured separately with a microsphere technique in the two regions of the rat pancreas perfused by the superior mesenteric artery (SMA) or celiac artery (CA) respectively. Intravenous infusion of CRF (0.25 microgram/kg b.w./min) caused an increase in both whole pancreatic blood flow and islet blood flow in the region of the pancreas perfused by the SMA. The fraction of whole pancreatic blood flow diverted through the islets in this part of the pancreas was, however, unaffected by CRF infusion (approximately 10%). CRF did not change either pancreatic or islet blood flow in the CA-perfused part of the pancreas, and did not affect the release of insulin.     

Effect of glutamine on protein synthesis in isolated intestinal epithelial cells

The purpose of this study was to determine the etiologic factors of denture stomatitis. Fifteen subjects with clinical evidence of localized simple denture stomatitis, fifteen subjects without clinical signs of denture stomatitis, and forty-five subjects with clinical evidence of generalized simple denture stomatitis were investigated clinically and mycologically. Subjects were evaluated according to age, sex, duration of denture usage, smoking habits, frequency of denture brushing, overnight denture wearing, pH level of saliva and degree of candidal colonization and candidal formation. Salivary samples and swabs were taken from the palate and the mucosal surfaces of the dentures investigated mycologically in order to identify the yeast colonies. Smears were taken from the palate and investigated in order to identify candidal formation. No statistically significant relationship was found between denture stomatitis and age, sex, duration of denture usage, frequency of denture brushing, overnight denture wearing or pH level of saliva. There was however, a statistically significant relationship between denture stomatitis and denture hygiene, smoking habits, candidal colonization and candidal formation.     

Lack of effects of some individual polybrominated diphenyl ether (PBDE) and polychlorinated biphenyl (PCB) congeners on human lymphocyte functions in vitro

The structural similarities between polybrominated diphenyl ethers and immunotoxic halogenated aromatic compounds suggest that the polybrominated diphenyl ethers might affect the immune system. The present study was undertaken to investigate the immunological effects of some purified PBDE-congeners on human lymphocyte function in vitro. Polychlorinated biphenyl congeners were also included in the study. Mitogen-induced DNA synthesis and immunoglobulin synthesis by lymphocytes from blood donors were examined following polybrominated diphenyl ether or polychlorinated biphenyl exposure in vitro in order to determine the immunotoxic potential of these substances. No effects on mitogen-induced proliferation or immunoglobulin synthesis were observed after exposure of cells to concentrations up to 10(-5) M. The negative findings in this study indicate that certain functions of human peripheral lymphocytes, i.e. proliferation and immunoglobulin synthesis, are insensitive to the direct action of polybrominated diphenyl ethers and polychlorinated biphenyls. Our results are in accordance with other recent studies in which no effects on immunological parameters were demonstrated by exposure of lymphocytes to polyhalogenated aromatic hydrocarbons in vitro.     

Identification of N,N'-dicyclohexylcarbodiimide-reactive glutamic and aspartic acid residues in Escherichia coli transhydrogenase and the exchange of these by site-specific mutagenesis

Pyridine nucleotide transhydrogenase (EC 1.6.1.1) from Escherichia coli was investigated with respect to the role of glutamic and aspartic acid residues reactive to N,N'-dicyclohexylcarbodiimide (DCCD) and potentially involved in the proton-pumping mechanism of the enzyme. The E. coli transhydrogenase consists of an alpha (510 residues) and a beta (462 residues) subunit. DCCD reacts with the enzyme to inhibit catalytic activity and proton pumping. This reagent modifies Asp alpha 232, Glu alpha 238, and Glu alpha 240 as well as amino acid residue(s) in the beta subunit. Using the cloned and overexpressed E. coli transhydrogenase genes (Clarke, D. M., and Bragg, P. D. (1985) J. Bacteriol. 162, 367-373), Asp alpha 232 and Glu alpha 238 were replaced independently by site-specific mutagenesis. In addition, Asp alpha 232, Glu alpha 238, and Glu alpha 240 were replaced to generate triple mutants. The specific catalytic activities of the mutant transhydrogenases alpha D232N, alpha D232E, alpha D232K, alpha D232H, alpha E238K, and alpha E238Q as well as of the triple mutants alpha D232N, alpha E238Q, alpha E240Q and alpha D232H, alpha E238Q, alpha E240Q were in the range of 40-90% of the wild-type activity. Proton-pumping activity was present in all mutants. Examination of the extent of subunit modification by [14C]DCCD revealed that the label was still incorporated into both alpha and beta subunits in the Asp alpha 232 mutants, but that the alpha subunit was not labeled in the triple mutants. Catalytic and proton-pumping activities were nearly insensitive to DCCD in the triple mutants. This suggests that loss of catalytic and proton-pumping activities is associated with modification of the aspartic and glutamic acid residues of the alpha subunit. In the presence of the substrate NADPH, the rate of modification of the beta subunit by [14C]DCCD was increased, and there was a greater extent of enzyme inactivation. By contrast, NADH and 3-acetylpyridine-NAD+ protected the catalytic activity of the transhydrogenase from inhibition by DCCD. The protection was particularly marked in the E238Q and E238K mutants. It is concluded that the Asp alpha 232, Glu alpha 238, and Glu alpha 240 residues are not essential for catalytic activity or proton pumping. The inactivation by DCCD is likely due to the introduction of a sterically hindering group that reacts with the identified acidic residues close to the NAD(H)-binding site.