Analysis of glycosyl bond cleavage and related isotope effects in collision-induced dissociation quadrupole-time-of-flight mass spectrometry of isomeric trehaloses

The configuration isomers alpha,alpha-, alpha,beta-, and beta,beta-trehalose are distinguishable by a relative ion abundance analysis using collision-induced dissociation MS/MS measurements in electrospray ionization quadrupole-time-of-flight mass spectrometry. The relative abundance of the Y-type fragment ion of alpha,alpha-trehalose is the highest and that of beta,beta-trehalose is the lowest, indicating that alpha-glycosyl bonds cleave more easily than beta-glycosyl bonds. The relative ion abundance depends on both the alpha- and beta-glycosyl linkage type and the number of alpha-glycosyl bonds. The reaction path of glycosyl bond cleavage is calculated computationally using the molecular orbital method in the form of Hartree-Fock theory in conjunction with the 6-31G(d) basis set. The results are consistent with the experimental data. Isotope effects on the fragmentation of the glycosyl bonds are detected in the experiments of the H2O/D2O solvent systems. Furthermore, the isotope effect regarding beta,beta-trehalose is larger than those of alpha,alpha- and alpha,beta-trehalose, indicating that the isotope effect on the beta-glycosyl bond cleavage is larger than that on the alpha-glycosyl bond cleavage. The thermal energy increase in trehalose-d8 molecules over the corresponding trehalose molecules is calculated from the vibrational modes.     

Development of multianode photomultiplier tube

We have developed a multianode photomultiplier tube (MAPMT) which has a sensitive cathode diameter of 36 mm and an anode with 88 segments. The position sensitivity of the tube was studied by using light from a LED and scintillation light from a scintillation-fiber bundle. We observed particle tracks for the first time by using a single photomultiplier tube.     

Altered response to histamine in brain tumor vessels: the selective increase of regional cerebral blood flow in transplanted rat brain tumor

The authors studied the effect of intracarotid administration of histamine on the regional cerebral blood flow (rCBF) in transplanted rat C6 glioma by the hydrogen clearance method. Histamine infusion at doses of 1 and 10 micrograms/kg/min produced an increase of rCBF in the tumor (24.6% +/- 16.4%, p < 0.002, and 37.6% +/- 18.2%, p < 0.0001, respectively) and also in brain surrounding the tumor (26.8% +/- 16.2%, p < 0.002, and 34.9% +/- 9.2%, p < 0.0001, respectively) without any significant changes in the ipsilateral hemisphere. Intravenous administration of pyrilamine (H1 antagonist) and cimetidine (H2 antagonist) reduced blood flow responses to histamine; cimetidine was a more effective blocking agent than pyrilamine. Intracarotid infusion of histamine (1 and 10 micrograms/kg/min) with intravenous injection of Evans blue dye disclosed the selective extravasation of dye in the tumor and the brain surrounding the tumor. These results indicated that brain tumor vessels could respond to histamine differently than normal brain capillaries. The mechanism of selective response to histamine could be explained either by increased permeability or by altered characteristics of histamine receptors in the tumor vessels.     

Evidence that a phosphatidylinositol 3,4,5-trisphosphate-binding protein can function in nucleus

PIP3BP is a phosphatidylinositol 3,4,5-trisphosphate-binding protein (PIP3BP) abundant in brain, containing a zinc finger motif and two pleckstrin homology (PH) domains. Staining of rat brain cells with anti-PIP3BP antibody and determination of localization of PIP3BP fused to the green fluorescent protein (GFP-PIP3BP) revealed that PIP3BP was targeted to the nucleus. Targeting was dependent on a putative nuclear localization signal in PIP3BP. Generation of PIP3 in the nucleus was detected in H2O2-treated 293T cells, nerve growth factor (NGF)-treated PC12 cells, and platelet-derived growth factor (PDGF)-treated NIH 3T3 cells. Translocation of phosphatidylinositol 3-kinase (PI 3-kinase) to the nucleus and enhanced activity of PI 3-kinase in the nucleus fraction were observed after H2O2 treatment of 293T cells, suggesting that PI 3-kinase can be activated in the nucleus as well as in the membrane after appropriate stimulation of the cells. Co-expression of the constitutively active PI 3-kinase with PIP3BP resulted in exportation of the protein from the nucleus to the cytoplasm, suggesting that PIP3BP can function as a PIP3-binding protein in the intact cells. These results imply that there may be an unknown function of PI 3-kinase in the nucleus.     

Effective and Green Synthesis of Methyl Pyrrole-1-carboxylate with Dimethyl Carbonate Over Solid Base

The methyl pyrrole-1-carboxylate (1-MPC) was synthesized by highly selective one-pot N-methoxycarbonylation of pyrrole with dimethyl carbonate (DMC). Solid bases were found to be effective catalysts and the formation of 1-MPC was closely related to their basicity. The in-situ FTIR indicated that the role of solid base catalysts was to activate pyrrole via the formation of O–H bond and the weakening of N–H bond, which was attributed to the interaction between pyrrole and surface oxygen anion of solid base. Furthermore, the reaction temperature and the reactants molar ratio were found to be the main factors for the improvement of pyrrole conversion.     

Large-Scale Shake Table Experiment and Numerical Simulation on the Nonlinear Behavior of Pile-Groups Subjected to Large-Scale Earthquakes

This paper describes the results of large-scale shake-table experiments involving a 3×3 pile-group. The pile-group was embedded in dry sand and subjected to sinusoidal waves and an earthquake motion recorded from the 1995 Hyogo-ken Nanbu (Kobe) earthquake. The load transfer between soil and pile was derived and the group effect was captured. Numerical simulations were also performed using a Beam-on-Nonlinear-Winkler-Foundation approach with a new hysteretic p-y curve. A comparison of the experimental and numerical results revealed that the numerical simulation is capable of accounting for the soil-pile interaction observed in the experiment.     

Discrepancy between provocative test and clinical results following endovascular obliteration of spinal arteriovenous malformation

The isolated and perfused kidney of the spontaneously hypertensive rat (SHR) exhibits an increased vascular reactivity to serotonin when compared to normotensive Wistar-Kyoto (WKY) rats. Experiments were designed to determine the involvement of a prostanoid constricting factor in the augmented response to serotonin in the SHR kidney. Kidneys taken from male (12 months) SHR and WKY rats were studied in parallel and perfused with Tyrode's solution at constant, optimal flow rates. Vasoconstrictor response were recorded as increases in perfusion pressure. The dose-response curves to serotonin obtained in the SHR were shifted to the left compared to the WKY. Indomethacin decreased the responses to the smaller doses of serotonin in the SHR, but increased those to the higher doses of the monoamine in the WKY. The responsiveness to the monoamine was no longer significantly different in the two strains in the presence of the inhibitor of cyclooxygenase. Dazoxiben, an inhibitor of thromboxane synthetase, did not alter the responses to serotonin in either the WKY and SHR kidneys. These experiments suggest that a prostanoid, but not thromboxane A2, may play a role in the augmented response to serotonin in the kidney of aged SHR.     

Effects of buprenorphine and Ro 15-4513 on delayed death and brain beta-endorphin levels in rats treated with cocaine or cocaine-ethanol

The present study was aimed at elucidating the relationship between brain beta-endorphin, which was estimated by the immunofluorescence method, and fatal drug toxicities due to cocaine and combined cocaine-ethanol administration, including the late fatal toxicities clinically noted. beta-endorphin is an endogenous opioid peptide, and its secretion has been suggested to be influenced by physiological stresses. Furthermore, since protection against these fatal toxicities has been previously reported to be provided by buprenorphine (a ligand for opioid receptors) and Ro 15-4513 (a ligand for benzodiazepine receptors), this study also focused on the relationship between the effects of these two ligands and the changes in brain beta-endorphin immunoreactivity. In the fatal toxicity study, a toxic dose (75 mg/kg, i.p.) of cocaine combined with and without ethanol (3 g/kg, i.p.) was administered to the rats, with and without buprenorphine (0.25, 0.5, 1 mg/kg, i.p.) or Ro 15-4513 (5, 10, 15 mg/kg, i.p.). All of the deaths that occurred in these animals were divided into two groups: early deaths with early toxic symptoms in which the drugs were detected in the tissue samples, and late deaths with late toxic symptoms in which no drugs were detected in the samples. Without the administration of buprenorphine or Ro 15-4513, the frequency of late deaths was higher in the cocaine group as compared to the cocaine-ethanol group. The total mortality rate was effectively attenuated by treatment with 0.25 mg/kg buprenorphine or 10 mg/kg Ro 15-4513. Following treatment with 1 mg/kg buprenorphine or 15 mg/kg Ro 15-4513, the frequency of late deaths was significantly enhanced in the cocaine group. The brain and liver cocaethylene concentrations were also attenuated in those groups in which the total mortality rates were attenuated. In the brain beta-endorphin immunoreactivity study, the number of beta-endorphin immunoreactive nerve cells at the arcuate nucleus was counted at 3 minutes or 24 hours after the drug treatment. At 3 minutes after the drug treatment, the number of weakly immunoreactive cells with photographic light absorption values greater than 50% was enhanced in the groups in which the frequency of late deaths had been increased. In the cocaine-ethanol groups treated with buprenorphine or Ro 15-4513, this enhancement of weakly immunoreactive cells was observed when the total mortality rate was increased, regardless of the type of death. At 24 hours after the drug treatment (50 mg/kg cocaine), an enhancement of the weakly immunoreactive cells only was observed in all of the groups in which the occurrence of toxicities had been enhanced, regardless of the type of toxicity. Therefore, it can be concluded that the enhancement of total brain beta-endorphin immunoreactivity was closely correlated with the increase in the frequency of total fatal toxicities, and that the enhancement of weakly immunoreactive cells was closely correlated with the increase in the frequency of delayed fatal toxicities.