Biomedical subjects

T Maruyama

Publications and source records attributed to T Maruyama.

At least 127 records · Page 7Linked to original sources

PubMed2001

Small heat shock protein of a hyperthermophilic archaeum, Thermococcus sp. strain KS-1, exists as a spherical 24 mer and its expression is highly induced under heat-stress conditions.

Small heat shock proteins (sHsps) are the most ubiquitous molecular chaperones. Several sHsps have been shown to exhibit chaperone activity and protect proteins from thermal and chemical aggregation. We have characterized a small heat shock protein from a hyperthermophilic archaeum, Thermococcus sp. strain KS-1. Electron microscopy revealed that the protein exists as a spherical oligomer with a diameter of 14+/-1 nm. The molecular weight of the oligomer was determined to be 478.6 kDa by size exclusion chromatography-multiangle laser light scattering. Thus, the Thermococcus sHsp is likely to exist as a spherical 24meric oligomer with almost the same structure as the Methanococcus jannaschii sHsp. The Thermococcus sHsp homo-oligomer protected porcine heart citrate synthase from thermal aggregation. It also slightly enhanced the refolding of acid-denatured green fluorescent protein. While the Thermococcus sHsp could not be detected in cells grown at the optimal growth temperature or lower, the expression of the protein was highly induced when the cells were grown at temperatures higher than the optimal growth temperature. Since only group II chaperonins and sHsps exist in hyperthermophilic archaea as molecular chaperones, sHsps should have an important role in protecting cells from lesions caused by aggregates of thermally denatured cellular proteins.

Journal Article↗

PubMed2001

Effect of hydrocarbon-water interfaces on synthetic and hydrolytic activities of lipases.

We have developed a new method of lipase activation for interesterification, in which lipase, in contact with a hydrocarbon-water interface, has been found to have high interesterification activity in an anhydrous solvent. We have applied this activation method to various lipase and obtained high synthetic activity in n-hexane, and have investigated the effect of various hydrocarbon-water interfaces on the synthetic and hydrolytic activities of lipases. The esterification and/or interesterification activity of lipases tested was improved by this activation method, using an n-tetradecane-water interface. From the initial group of lipases, three representative lipases (from Rhizopus japonicus, Chromobacterium viscosum and porcine pancreas) were selected for further study. The effect of various hydrocarbon-water interfaces on synthetic (interesterification or esterification) activity was studied. We demonstrated that the resulting synthetic activity was affected by the choice of hydrocarbon-water interface and that there were differences in the effects of interfaces on the synthetic activity of these lipases.

Journal Article↗

PubMed2001

Small-angle X-ray scattering analysis of stearic acid modified lipase.

Stearic acid modified lipase (from Rhizopus japonicus) exhibited remarkable interesterification activity in n-hexane, but crude native lipase did not. The structure of the fatty acid modified lipase had not been analyzed until now. We analyzed the modified lipase by small-angle X-ray scattering (SAXS) measurements in order to clarify the structure. SAXS measurements showed that the modified lipase consisted of a lipid lamellar structure and implied that the lipase was incorporated into the lamellar structure of stearic acid. The long spacings in the lamellar structures of the modified lipase and stearic acid were measured.

Algorithms↗

PubMed2001

A cross-protection experiment in pigs vaccinated with Haemophilus parasuis serovars 2 and 5 bacterins, and evaluation of a bivalent vaccine under laboratory and field conditions.

Cross-protection between Haemophilus parasuis serovars 2 and 5 was examined in pigs using a bacterin based vaccine, and subsequently the safety and efficacy of a bivalent vaccine were evaluated. Upon intratracheal challenge of a serovar 2 or 5 strain, pigs immunized with a monovalent vaccine were protected against challenge with a homologous serovar strain, but not with a heterologous serovar strain. Immunization with a bivalent vaccine containing both serovars 2 and 5 bacterins conferred protection in pigs against lethal challenge with each of the serovar strains. A total of 86 pigs from two SPF herds were injected with the bivalent vaccine intramuscularly twice at a four-week interval. No adverse reactions following the vaccination were observed. On day 7 after the second vaccination, vaccinated and non-vaccinated control pigs from herd A were transferred to herd B, where Glasser's disease had broken out. Pigs in the control group developed clinical signs of the disease, and 6 of 8 (75%) pigs died until slaughter, in contrast with only 4 of 46 (9%) pigs in the vaccinated group. In herd C, where there was no outbreak of Glasser's disease, complement fixation antibody titer was raised only in the vaccinated group. A challenge experiment on days 20 and 79 after the second vaccination showed that only the vaccinated pigs were protected. From these findings, the safety and efficacy of the bivalent vaccine were confirmed under laboratory and field conditions.

Animals↗

PubMed2001

Effects of increased physical activity and mild calorie restriction on heart rate variability in obese women.

The effects of exercise and mild calorie restriction on heart rate variability (HRV) were investigated in 12 mildly obese, normotensive Japanese women aged 45.8+/-4.2 (SEM) years with a body mass index (BMI) of 27.3+/-0.4 kg/m2. The subjects participated in a 3-month program aimed at increasing physical activity and modifying eating behavior (intervention group). The control group consisted of 12 women (age 50.1+/-4.8 years, BMI 27.2+/-0.6 kg/m2) who did not attend the program. The frequency domain of HRV was calculated from 5-min Holter recordings while the subjects rested in a supine position. After 3 months, BMI decreased to 25.0+/-0.5 kg/m2 (p<0.001 vs baseline) in the intervention group, which was accompanied by decreases in body fat mass, waist circumference, serum total cholesterol and triglycerides, and improvement in insulin sensitivity. The mean and SD of the RR intervals, total power, and low and high frequency power of HRV significantly increased after the intervention, whereas no significant changes were seen for the controls. The changes in these HRV variables (calculated by subtracting the baseline values from the follow-up values) negatively correlated with the change in waist circumference, with the Pearson correlation coefficients being between -0.50 and -0.62 (p<0.05). A negative correlation was also seen between the changes in high frequency power and insulin resistance estimated by homeostasis model assessment (r=-0.49, p<0.05). The combination of exercise and mild calorie restriction led to changes in HRV indicative of an improvement in parasympathetic modulation.

Body Mass Index↗

PubMed2001

NeuroD/beta2 gene G-->A polymorphism may affect onset pattern of type 1 diabetes in Japanese.

OBJECTIVE: The majority of type 1 diabetes is considered to be autoimmune with, for the most part, abrupt development. However, type 1 diabetes with slow onset, or the so-called slowly progressive type 1 diabetes or latent autoimmune diabetes in adults, has been recently recognized and is considered to be autoimmune-related. Although some investigators tried to explain the difference in onset pattern by the genetic background, including HLA type, it has not been established thus far. We hypothesized that the difference in onset pattern may relate to regeneration or differentiation of pancreatic beta-cells, and we therefore focused on the NeuroD/BETA2 gene, which encodes a transcription factor for the insulin gene and beta-cell differentiation. RESEARCH DESIGN AND METHODS: We examined the NeuroD/BETA2 gene polymorphism in 105 Japanese type 1 diabetic patients and in 122 nondiabetic Japanese subjects in a case-control study, and we stratified the patients according to their onset pattern and islet-associated autoantibody positivity. RESULTS: Regardless of the existence of islet-associated autoantibody, we found a significant difference in A allele frequency between type 1 diabetic patients with acute-onset type and control subjects. However, no difference was found between type 1 slow-onset diabetic patients and control subjects. CONCLUSIONS: These results support our hypothesis that NeuroD/BETA2 may affect the ability of regeneration of beta-cells, leading to a difference in the onset pattern and clinical course of type 1 diabetes.

Adolescent↗

PubMed2001

No association of type 1 diabetes with a microsatellite marker for CTLA-4 in a Japanese population.

Susceptibility to insulin-dependent (type 1) diabetes mellitus is determined by both environmental and genetic factors. The primary gene associated with predisposition to type 1 diabetes is the human leukocyte antigen (HLA) class II gene (IDDM1). Recent studies have described linkage and association of type 1 diabetes to the cytotoxic T lymphocyte antigen-4 (CTLA-4) gene (IDDM12)in Caucasians. CTLA-4 is a candidate gene for T-cell-mediated autoimmune diseases because it is a negative regulator of T-cell proliferation. We investigated distribution of a CTLA-4 (AT)n microsatellite marker in 118 Japanese patients with type 1 diabetes and 195 control subjects. We also investigated association between this CTLA-4 gene polymorphism and GAD65 antibody positivity in 103 of the patients. CTLA-4 microsatellite marker loci were determined by polymerase chain reaction amplification of genomic DNA and resolution of the products on sequencing gels. GAD65 antibody was detected by radioligand binding assay. There was no significant difference in the distribution of CTLA-4 alleles between patients and controls, and no difference was observed in the prevalence of CTLA-4 alleles when GAD65 antibody-positive and -negative individuals with the type 1 diabetes were compared. The present study did not support an association between the CTLA-4 microsatellite marker and type 1 diabetes in our Japanese study population.

Abatacept↗

PubMed2001

Reaction of 7-(2-mesyloxy-2-phenylethyl)theophylline with amines: Synthesis of 1,2,3,6-tetrahydro-6-imino-2-oxo-7H-purine derivatives.

Theophylline was converted to 7-(2-phenyl-2-methanesulfonyloxy)ethyl congener and the product was treated with ammonia or primary amines in a mixture solution of water and organic solvents. Two products were proven to be the styrene analogue and 7-(2-amino-2-phenylethyl)theophylline. The structure of the third product was elucidated as the 1,2,3,6-tetrahydro-6-imino-2-oxo-7H-purine derivatives by spectroscopic analysis including HMBC correlation and X-ray crystallography.

Amines↗

PubMed2001

[Not Available].

Explore the source record for details and available documents.

Journal Article↗

PubMed2000

Quenching of singlet oxygen by carotenoids produced in escherichia coli - attenuation of singlet oxygen-mediated bacterial killing by carotenoids.

We examined the viability of Escherichia coli transformants harboring various carotenoids synthesizing genes in a medium containing an enzymatic singlet oxygen generating system, which contained myeloperoxidase, hydrogen peroxide and Br(-) at pH 4.5. Singlet oxygen quenching activities of various carotenoids in phosphatidyl choline micelles in aqueous media were also studied using the same enzymatic singlet oxygen generating system. Viability of the transformants producing carotenoids was higher than that of the wild type E. coli in the singlet oxygen generation mixture. Of the transformants tested, the viability of zeaxanthin-diglucoside producing transformant was the highest. Carotenoids in increasing order of k(q) values were beta-carotene, a cyclic carotene<zeaxanthin with hydroxy groups < or =lycopene, an acyclic carotene<canthaxanthin and astaxanthin with keto groups <<zeaxanthin-diglucoside. The k(q) value of zeaxanthin-diglucoside was 3.5 times higher than that of beta-carotene. These results suggest that orientation of the carotenoids in lipid layers of micelles and also in phospholipid membrane of bacteria is important for quenching of singlet oxygen. Furthermore, the viability of transformants producing lycopene and phytoene was almost as high as that of the transformant producing zeaxanthin-glucoside.

Adenosine Triphosphate↗

PubMed2000

Direct radiolabeling by [3H]quisqualic acid of group I metabotropic glutamate receptor in rat brain synaptic membranes.

[3H]Quisqualic acid (QA) was synthesized and used to label metabotropic glutamate receptor (mGluR) in rat brain synaptic membranes in the presence of three different ionotropic glutamate receptor agonists at respective saturating concentrations. Of several mGluR agonists tested, group I agonists were more potent in displacing [3H]QA binding than group II and group III agonists in the presence of the three ionotropic agonists. [3H]QA binding was markedly inhibited by guanine nucleotide analogues in a concentration-dependent manner at a concentration range of 10 nM to 1 mM. Scatchard analysis revealed that [3H]QA binding consisted of a single component with a K(d) of 50.9+/-5.3 nM and a B(max) of 431. 6+/-18.3 fmol/mg protein. These results suggest that [3H]QA indeed labels group I mGluR functionally coupled to GTP binding protein in rat brain synaptic membranes when determined under the experimental conditions employed.

Animals↗

PubMed2000

FK506-binding protein-type peptidyl-prolyl cis-trans isomerase from a halophilic archaeum, Halobacterium cutirubrum.

The halophilic archaeum, Halobacterium cutirubrum, has been shown to have a cyclophilin-type peptidyl-prolyl cis-trans isomerase (PPIase). Because most archaeal genomes studied only have genes for FK506-binding proteins (FKBPs) as a PPIase, it has been unclear whether H. cutirubrum has an FKBP-type PPIase or not. In the present study, a gene encoding an FKBP-type PPIase was cloned from genomic DNA of H. cutirubrum and then sequenced. This FKBP was deduced to be composed of 303 amino acid residues with a molecular mass of 33.3kDa. Alignment of its amino acid sequence with those of other reported FKBPs showed that it contained two insertion sequences in the regions corresponding to the bulge and flap of human FKBP12, which are common to archaeal FKBPs. Its C-terminal amino acid sequence was approximately 130 amino acids longer than the FKBPs of Methanococcus thermolithotrophicus and Thermococcus sp. KS-1. Among the 14 conserved amino acid residues that form the FK506 binding pocket, only three were found in this FKBP. This gene was expressed as a fusion protein with glutathione S-transferase (GST) in Escherichia coli, and the N-terminal GST portion was removed by protease digestion. The purified recombinant FKBP showed a weak PPIase activity with a low sensitivity to FK506. This FKBP suppressed aggregation of the unfolded protein.

Amino Acid Sequence↗

PubMed2000

Barnacle cement proteins. Importance of disulfide bonds in their insolubility.

Barnacles produce a cement that is a proteinaceous underwater adhesive for their secure attachment to the substratum. The biochemical properties of the cement have not previously been elucidated, because the insolubility of the cement proteins hampers their purification and characterization. We developed a non-hydrolytic method to render soluble most of the cement components, thereby allowing the proteins to be analyzed. Megabalanus rosa cement could be almost completely rendered soluble by its reduction with 0.5 m dithiothreitol at 60 degrees C in a 7 m guanidine hydrochloride solution, the high concentration of dithiothreitol being indispensable to achieve this. The effectiveness of this reduction treatment was confirmed by the detachment of the barnacle from the substratum. Three proteins comprising up to 94% of the whole cement were identified as the major cement components. The cDNA clone of one of these major proteins was isolated, and the site-specific expression of the gene in the basal portion of the adult barnacle, where the cement glands are located, was demonstrated. A sequence analysis revealed this cement component to be a novel protein of 993 amino acid residues, including a signal peptide. This is the first report of the major component of the barnacle cement protein complex.

Amino Acid Sequence↗

PubMed2000

Archaeal peptidyl prolyl cis-trans isomerases (PPIases).

PPIases are ubiquitous in living organisms. While 3 families of PPIases, cyclophilin (CyP), FK506 binding protein (FKBP) and parvulin (Pvn), have been studied in detail in Eukarya and Bacteria (eubacteria), little is known about archaeal PPIases. Among 2 cyclophilins found in Archaea, only Halobacterium cyclophilin (HcCyP19) has been characterized. It is a cyclosporin A (CsA) sensitive CyP with a MW of 19.4kDa. The PPIase activity and CsA sensitivity of this CyP is higher at higher salt concentration in the medium. No parvulin or its homolog has been found in Archaea. Two types of FKBPs, 26-30kDa long type and 17-18 kDa short type FKBP, have been found in Archaea. While the N-terminal regions of these 2 type FKBPs are similar to each other, the long type archaeal FKBP has an additional ca. 100 amino-acid sequence at its C-terminal region. In comparison with human HsFKBP12, the N-terminal region of the archaeal FKBP has 2 insertion sequences in the regions corresponding to Bulge and Flap of HsFKBP12. A short type archaeal FKBP from Methanococcus thermolithotrophicus has been shown to have not only a PPIase activity but also a chperone like activity, which includes protein refolding and aggregation suppressing activities with regard to protein folding intermediates. Mutational analysis revealed that this chaperone-like activity was independent of the PPIase activity, and that the insertion sequence in the region corresponding to the Flap seemed to be important.

Amino Acid Sequence↗

PubMed2000

Effects of the second-generation leukotriene B(4) receptor antagonist, LY293111Na, on leukocyte infiltration and collagen-induced arthritis in mice.

The effects of the second-generation leukotriene B(4) receptor (LTB(4) receptor) antagonist, 2-[2-propyl-3-¿3-[2-ethyl-4-(4-fluorophenyl)-5-hydroxy-phenoxy]-propo xy¿phenoxy]benzoic acid sodium salt (LY293111Na), on leukotriene B(4)-induced leukocyte infiltration and interleukin-1-accelerated collagen-induced arthritis in mice were studied. Neutrophil infiltration induced into an air pouch by leukotriene B(4) was dose-dependently inhibited by LY293111Na and strongly so by another LTB(4) receptor antagonist, 4-[5-¿4-(aminoiminomethyl)phenoxy¿pentoxy]-3-methoxy-N, N-bis(1-methylethyl) (Z)-2butenedioate (1:1) (CGS25019C). Both compounds significantly inhibited the increase of the arthritis index and the ankle bone destruction in interleukin-1-accelerated collagen-induced arthritis. Phenidone, a 5-lipoxygenase inhibitor, also inhibited interleukin-1-accelerated collagen-induced arthritis, while indomethacin and tenidap, cyclooxygenase inhibitors, had slight inhibitory effects. Injection of interleukin-1 elicited a marked increase of the leukotriene B(4) level in arthritic paws, while the prostaglandin E(2) level was slightly increased. These findings indicate clearly that leukotriene B(4) is an important mediator of interleukin-1-accelerated collagen-induced arthritis in mice. If this can be extrapolated to man, LTB(4) receptor antagonists might be useful for treatment of the acute progressive phase of human arthritis.

Animals↗

PubMed2000

Inhibitory effect of a prostaglandin E receptor subtype EP(1) selective antagonist, ONO-8713, on development of azoxymethane-induced aberrant crypt foci in mice.

We previously reported that prostaglandin E(2) contributes to colon carcinogenesis through its binding to the prostaglandin E receptor subtype EP(1) using a genetic approach in EP(1)-knockout mice and a pharmacological approach with the EP(1) selective antagonist, ONO-8711. In the present study, we examined the effects of another more selective EP(1) receptor antagonist, ONO-8713, on development of azoxymethane (AOM)-induced aberrant crypt foci (ACFs) in male C57BL/6J mice treated i.p. with 10mg/kg body weight AOM once a week for 3weeks. Administration of ONO-8713 at doses of 250, 500 and 1000ppm in diet during and post-AOM treatment for 5weeks resulted in a dose-dependent reduction of ACF formation, being 15, 30 and 36% inhibition of the control value, respectively. The level of inhibition was similar to that with ONO-8711. Moreover, ONO-8713 suppressed the development of ACF when administered at post-AOM, as in the case of ONO-8711. The data confirm EP(1) receptor involvement in colon carcinogenesis.

Animals↗

PubMed2000

Role of arg-410 and tyr-411 in human serum albumin for ligand binding and esterase-like activity.

Recombinant wild-type human serum albumin (rHSA), the single-residue mutants R410A, Y411A, Y411S and Y411F and the double mutant R410A/Y411A were produced using a yeast expression system. The recombinant proteins were correctly folded, as they had the same stability towards guanidine hydrochloride and the same CD spectrum as HSA isolated from serum (native HSA). Thus the global structures of the recombinant proteins are probably very similar to that of native HSA. We investigated, by ultrafiltration and CD, the high-affinity binding of two representative site II ligands, namely ketoprofen and diazepam. According to the crystal structure of HSA, the residues Arg-410 and Tyr-411 protrude into the centre of site II (in subdomain 3A), and the binding results showed that the guanidino moiety of Arg-410, the phenolic oxygen and the aromatic ring of Tyr-411 are important for ketoprofen binding. The guanidino moiety probably interacts electrostatically with the carboxy group of ketoprofen, the phenolic oxygen could make a hydrogen-bond with the keto group of the ligand, and the aromatic ring may participate in a specific stacking interaction with one of or both of the aromatic rings of ketoprofen. By contrast, Arg-410 is not important for diazepam binding. The two parts of Tyr-411 interact favourably with diazepam, and probably do so in the same way as with ketoprofen. In addition to its unique ligand binding properties, HSA also possesses an esterase-like activity, and studies with p-nitrophenyl acetate as a substrate showed that, although Arg-410 is important, the enzymic activity of HSA is much more dependent on the presence of Tyr-411. A minor activity could be registered when serine, but not alanine or phenylalanine, was present at position 411.

Amino Acid Sequence↗

PubMed2000

Prevalence and contamination levels of Listeria monocytogenes in retail foods in Japan.

Retail foods in Japan were surveyed for the presence and contamination levels of L. monocytogenes. It was isolated from 12.2, 20.6, 37.0 and 25.0% of 41 minced beef, 34 minced pork, 46 minced chicken and 16 minced pork-beef mixture samples, respectively. MPN values were higher than 100/g in five (10.9%) minced chicken samples, but lower than 100/g in all minced beef, pork and pork-beef mixture samples. The organism was also isolated from 5.4% of the 92 smoked salmon samples at MPN values lower than 10/g, and from 3.3% of 213 ready-to-eat raw seafood samples at MPN values from lower than 0.3 to higher than 100/g. None of the 285 vegetable samples were contaminated with L. monocytogenes. These findings indicate that ready-to-eat raw seafoods are relatively high risk among the foods surveyed in this study.

Animals↗