A Novel Reaction of Sugars with Anion Radical of Carbon Dioxide Produced from Formate

Radiolysis of some monosaccharides (fructose, glucose and ribose) in air-free condition was markedly enhanced by the addition of formate at concentrations above 20 mm, while it was inhibited at concentrations below 20 mm. The following compounds were detected in the irradiated sugar solutions containing excess formate (100mm): 1-Deoxy-d-arabinohexulose (1, G=4.4) and 1,3- dideoxy-d-erythrohexulose (2, G= 1.3) from fructose; 2-deoxy-d-ribose (3, G=2.3) and 2-deoxyribitol (4, G =0.6) from ribose; and 2-deoxy-d-glucose (5, G=0.5) and 2-deoxy-d-glucitol (6, G=0.4) from glucose. A mechanism for radiolytic formation of the products was proposed, based on interaction of - formed from formate with sugars.     

Red Pigment Formation by the Reaction of Oxidized Ascorbic Acid and Protein in a Food Model System of Low Moisture Content

Ascorbic acid-protein mixtures of low moisture content stored for 2~3 days in aerobic conditions at 60°C produced a red coloration, which was shown to have resulted from an amino-carbonyl reaction of oxidized ascorbic acid (dehydroascorbic acid, DHA) by the facts that DHA-casein or ovalbumin systems yielded a rapid red coloration in low moisture conditions as well as in an ethanol suspension. Zein showed only a weak red coloration with DHA. An apparent decrease in the free amino group, as determined by the TNBS method, was observed to be associated in parallel with the red pigment formation. The electrophoretic pattern of ovalbumin changed significantly upon incubation with DHA, together with the formation of the red pigment. The red pigment extracted from a DHA-casein system showed an absorption spectrum, TLC Rf value, and hydrolyzed products (DHA and scorbamic acid) identical to those of 2,2′-nitrilo di-2(2)-deoxy-l-ascorbic acid mono ammonium salt, produced from DHA and amino acid. Formation of the red pigment was also observed in the reaction of DHA with N_α-acetyl lysine. These results indicate that the ε-amino group of lysine in protein can be attributed to the formation of a red pigment identical with NDA.     

Use of lac Gene Fusions to Study Regulation of Tyramine Oxidase,Which is Involved in Derepression of Latent Arylsulfatase in Escherichia coli

Strains with lac fused to each of the arylsulfatase (ats) and tyramine oxidase (tyn) operons in Escherichia coli were isolated. Synthesis of β-galactosidase in strains with tyn:: lac fusions was induced by tyramine, histamine, tryptamine, dopamine and octopamine, and the induction of the tyn operon was subject to catabolite and ammonium repressions. These repressions were relieved when the cells were grown with a poor carbon or nitrogen source. No arylsulfatase activity is detected in E. coli strains. Synthesis of β-galactosidase in strains with ats:: lac fusions was repressed by sulfur compounds. The repression was relieved by monoamine compounds, which induced tyramine oxidase synthesis. The inhibition of tyramine oxidase activity by cysteine resulted in a decrease of the derepressed synthesis of β-galactosidase in the ats:: lac fusion. Repressing and derepressing conditions for the tyn operon prevented and stimulated, respectively, expression of the ats operon. Thus, the expression of latent arylsulfatase in E. coli seems to be regulated by expression of the tyn operon.     

Study on the Correlation of Chemical Structure and Ovicidal Activities of 2-Bromoethylthiobenzenes

Formation of a sulfonium-like intermediate was assumed in the hydrolysis of the 2-bromoethylthiobenzenes. A linear free energy relationship was found between the hydrolysis rate of a certain substituted 2-bromoethylthiobenzene and the molar fraction of water in the solvent. The effect of the substituent on the rate constant was attributed not only to the activation energy but also to the entropy change of activation. The negative ρ-value in the formation of the sulfonium-like intermediate in aqueous solution was comparable with that obtained in the ρ-σ-π analysis for ovicidal activity of the compounds.

For the reaction of the substituted 2-bromoethylthiobenzenes with highly excess amount of 4-(p-nitrobenzyl)-pyridine, the ρ-value was found to be negative, which means that the formation of the sulfonium-like intermediate is a rate determining step. Whichever might be more important, the hydrolysis or alkylation, as to the ovicidal action of the compounds, the formation of the sulfonium-like intermediate, could be considered to be an essential step.     

Formation of Nicotinamide Ribose Diphosphate Ribose,a New Metabolite of NAD,by Aspergillus niger

Aspergillus niger (AKU 3302) degraded NAD to form Compound X. This compound was identified as nicotinamide ribose diphosphate ribose (NAmRDPR) by hydrolysis with alkaline or phosphodiesterase followed by chemical analysis of the products. NAmRDPR showed absorption maxima at 265~266 nm in 0.1 n HCI and 325 nm in 1.0 n KCN. Optimal pH for NAmRDPR formation by the enzyme preparation from this organism was around 4.0. Formation of NAmRDPR proceeded stoichiometrically with degradation of NAD. Some of other strains of A. niger formed NAmRDPR, but production of this compound was not demonstrated in other mold genera.     

Decomposition of Allyl Isothiocyanate in Aqueous Solution

Allyl isothiocyanate was gradually decomposed in aqueous solution to produce a garlic like odor. The decomposition of this isothiocyanate was not based on hydrolysis of R-NCS as in the case of p-hydroxybenzyl isothiocyanate, but the addition reaction on –N=C=S. Four substances formed with the decomposition of the isothiocyanate were isolated, and their chemical constitutions were clarified. Allyl isothiocyanate was decomposed to allyl allyldithiocarbamate (III), which was degradated to diallyl tetra- and penta-sulfide (II), and this poly-sulfide was further degradated to paraffin like hydrocarbon (I) and sulfur. Moreover, N,N′-diallylthiourea (IV) was produced by the addition reaction of allylamine, formed from the isothiocyanate by action of water, to residual isothiocyanate.     

Exposure and risk assessment of 1,3-butadiene in Japan

1,3-Butadiene is on the list of Substances Requiring Priority Action published by the Central Environmental Council of Japan in 1996. Emission of 1,3-butadiene has been controlled by a voluntary reduction program since 1997. Although the industrial emission of 1,3-butadiene in Japan has decreased in recent years, primarily due to a voluntary industrial emissions reduction program, the risks of exposure to it remain largely unknown. We assessed the risks and consequences of exposure to 1,3-butadiene on human health. A remarkable advantage of our risk assessment approach is the detailed assessment of exposure. Previously, we developed two different models that can be applied for the assessment of exposure: the first, the AIST-ADMER model estimates regional concentration distributions, whereas the second, the METI-LIS model estimates concentration distributions in the vicinity of factories. Both models were used for the assessment of exposure to 1,3-butadiene. Using exposure concentration and carcinogenic potency determined and reported by Environment Canada and Health Canada, we evaluated the excess lifetime cancer risk for persons exposed to 1,3-butadiene over the course of a lifetime. The results suggested that the majority of the population in Japan has an excess lifetime cancer risk of less than 10(-5), whereas a small number of people living close to industrial sources had a risk of greater than 10(-5). The results of the present assessment also showed that 1,3-butadiene in the general environment originates primarily from automobile emissions, such that a countermeasure of reducing emissions from cars is expected to be effective at reducing the total cancer risk among Japanese. On the other hand, individual risks among a population living in the vicinity of certain industrial sources were found to be significantly higher than those of the population living elsewhere, such that a reduction in emissions from a small number of specific industrial sources should be realized in order to reduce the high level of individual risk. Based on the results of our assessment, the Industrial Structure Council of the Ministry of Economy, Trade and Industry (METI) in Japan decided to announce that the voluntary reduction program had been successful, and that emissions reductions should no longer be targeted across all industries in general, but instead that such reductions should be carried out in a small number of selected factories that emit excessively large amounts of emissions.     

Accumulation of 8-oxo-deoxyguanosine in cardiovascular tissues with the development of hypertension

Accumulation of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dG) in DNA is associated with mutagenesis and cell death. Little attention has been given to the biological significance of 8-oxo-dG accumulation in cardiovascular tissues during the different stage of hypertension and its prevention. We thus investigated the levels and localization of both 8-oxo-dG accumulation and expression of MTH1, which hydrolyzes 8-oxo-dGTP to prevent its incorporation into DNA, in the thoracic aorta prepared from stroke-prone spontaneously hypertensive rats (SHRSP) and age-matched Wister-Kyoto rats (WKY), aged 5-32 weeks. HPLC-MS/MS analysis revealed that the levels of nuclear 8-oxo-dG in the aorta increased significantly in SHRSP, but not WKY, with aging. Immunohistochemical study revealed that both TUNEL reactivity and 8-oxo-dG immunoreactivity were increased in smooth muscle cells (SMC) and endothelial cells (EC) of the aorta with aging, and they exhibited similar distributions in serial sections. The number of 8-oxo-dG and TUNEL positive cells in EC, but not in SMC, was significantly higher in SHRSP than WKY at 32 weeks of age. In contrast, the expression levels of Mth1mRNA and MTH1 protein in the aorta were similarly decreased both in SHRSP and WKY with aging. However, the number of MTH1 expressing EC was remarkably increased in the older SHRSP compared to the younger ones or age-matched WKY. Hypertension significantly increased not only 8-oxo-dG accumulation but also the expression of MTH1 in EC of the aorta during aging. While accumulation of 8-oxo-dG in SMC of the aorta was slightly increased, the expression of MTH1 protein in SMC was rather decreased by hypertension. We thus suggest that MTH1 may protect EC in the aorta from the oxidative damage increased by hypertension.     

14-3-3 is involved in p75 neurotrophin receptor-mediated signal transduction

The low affinity neurotrophin receptor (p75NTR) has been shown to mediate the apoptosis signaling to neural cells. However, the specific mechanisms of intracellular signal transduction of this process are largely unknown. To understand p75NTR-mediated signal transduction, we previously identified a protein that interacts with the intracellular domain of p75NTR, and we named it p75NTR-associated cell death executor (NADE). To elucidate further the signaling mechanisms utilized by p75NTR and NADE, we screened for NADE-binding protein(s) with the yeast two-hybrid method, and we identified 14-3-3epsilon as a NADE-binding protein in vivo. To examine whether 14-3-3epsilon affects the induction of p75NTR-mediated apoptosis, wild type or various deletion mutant forms of 14-3-3epsilon were co-expressed in HEK293, PC12nnr5, and oligodendrocytes. Interestingly, transient expression of the mutant form of 14-3-3epsilon lacking the 208-255 amino acid region blocked nerve growth factor-dependent p75NTR/NADE-mediated apoptosis, although this mutant form of 14-3-3epsilon continued to associate with NADE. These results suggest that 14-3-3epsilon plays an important role in the modulation of nerve growth factor-dependent p75NTR/NADE-mediated apoptosis.     

Evaluation of a high-affinity QCM immunosensor using antibody fragmentation and 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer

This study evaluated construction of a highly affinitive quartz crystal microbalance (QCM) immunosensor using anti-C-reactive protein (CRP) antibody and its fragments for CRP detection. Three types of antibody were immobilized on the surface of a QCM via covalent-bounding. Then affinity was evaluated through antigen-antibody binding between CRP and its antibody. Affinity between antigen-antibody was shown to be highest when anti-CRP F(ab')2-IgG antibody (70 microg/mL) was immobilized on the QCM. In case of anti-CRP F(ab')2-IgG antibody, affinity which was attributable to antigen-antibody binding was almost twice that of anti-CRP IgG antibody, which is used conventionally for QCM immunosensors. In addition, when it was treated with 2-methacryloyloxyethyl phosphorylcholine-co-n-butyl methacrylate, so-called MPC polymer, highly affinitive and selective immunosensing for CRP was achieved without non-specific binding from plasma proteins in human serum. When anti-CRP F(ab')2-IgG antibody was immobilized on the QCM, the detection limit and the linearity of CRP calibration curve were achieved at concentrations from 0.001 to 100 microg/dL even during investigation in serum samples. Experimental results verified the successful construction of a highly affinitive and selective QCM-immunosensor which was modified with anti-CRP F(ab')2-IgG antibody and MPC polymer.