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Biomedical subjects

Hiroyuki Ohno

Publications and source records attributed to Hiroyuki Ohno.

At least 37 records · Page 2Linked to original sources
PubMed2005

Characterization of carbohydrate-binding cytochrome b562 from the white-rot fungus Phanerochaete chrysosporium.

cDNA encoding a hemoprotein similar to the cytochrome domain of extracellular flavocytochrome cellobiose dehydrogenase (CDH) was cloned from the white-rot fungus Phanerochaete chrysosporium. The deduced amino acid sequence implies that there is a two-domain structure consisting of an N-terminal cytochrome domain and a C-terminal family 1 carbohydrate-binding module (CBM1) but that the flavin-containing domain of CDH is not present. The gene transcripts were observed in cultures in cellulose medium but not in cultures in glucose medium, suggesting that there is regulation by carbon catabolite repression. The gene was successfully overexpressed in Pichia pastoris, and the recombinant protein was designated carbohydrate-binding cytochrome b562 (CBCyt. b562). The resonance Raman spectrum suggested that the heme of CBCyt. b562 is 6-coordinated in both the ferric and ferrous states. Moreover, the redox potential measured by cyclic voltammetry was similar to that of the cytochrome domain of CDH. These results suggest that the redox characteristics may be similar to those of the cytochrome domain of CDH, and so CBCyt. b562 may have an electron transfer function. In a binding study with various carbohydrates, CBCyt. b562 was adsorbed with high affinity on both cellulose and chitin. As far as we know, this is the first example of a CBM1 connected to a domain without apparent catalytic activity for carbohydrate; this CBM1 may play a role in localization of the redox protein on the surface of cellulose or on the fungal sheath in vivo.

Amino Acid Sequence↗
PubMed2005

[Headspace GC/MS analysis of residual vinyl chloride and vinylidene chloride in polyvinyl chloride and polyvinylidene chloride products].

A headspace GC/MS analysis method for the simultaneous determination of residual vinyl chloride (VC) and vinylidene chloride (VDC) in polyvinyl chloride (PVC) and polyvinylidene chloride (PVDC) products was developed. A test sample was swelled overnight with N,N-dimethylacetamide in a sealed vial. The vial was incubated for 1 hour at 90 degrees C, then the headspace gas was analyzed by GC/MS using a PLOT capillary column. The recoveries from spiked PVC and PVDC samples were 90.0-112.3% for VC and 85.2-108.3% for VDC. The determination limits were 0.01 microg/g for VC and 0.06/microg/g for VDC, respectively. By this method, VC was detected in two PVC water supply pipes at the levels of 0.61 and 0.01 microg/g. On the other hand, VC and VDC were not detected in any of the food container-packages or toys tested.

Dichloroethylenes↗
PubMed2005

[Indoor air pollution due to 2-ethyl-1-hexanol airborne concentrations, emission sources and subjective symptoms in classroom users].

OBJECTIVE: 2-Ethyl-1-hexanol (2E1H) is a volatile organic compound (VOC) which seldom attracts attention in Japan. This study aimed at clarifying changes in its concentration over time, emission sources, and students' symptoms in classrooms of a university building where indoor air was found to be markedly polluted with 2E1H. METHODS: From March 2001 through September 2002, we measured VOC concentrations in Building A, constructed in 1998, as well as Building B (Sept. 2002), constructed over 30 years ago and considered as a control. Airborne concentrations of 13 carbonyl compounds were quantified with diffusive samplers and high-performance liquid chromatography, and those of 41 other VOCs with an active sampling method using charcoal tubes and a gas chromatograph with a mass spectrometer (GC-MS). In August 2002, we also measured VOC emissions from the floors using double-cylinder chambers and the airborne concentrations of phthalate esters by filtration sampling, both by GC-MS. Subjective symptoms in 315 student classroom users in Building A and 275 in Building B were surveyed in July 2002 with anonymous self-administered questionnaires. RESULTS: 2E1H concentrations in Building A, which exceeded the Japanese recommended threshold of total VOCs (400 microg/m3) in some measurements, tended to be lower in winter and higher in summer, and did not show any tendency for decrease over time. No association was found between indoor concentrations of phthalate esters and those of 2E1H. The concentrations clearly differed between rooms, related to emission rates from the floors. Carpeting materials had been placed directly on the concrete floors in rooms with higher emission levels, whereas the carpeting materials and the concrete floor did not make contact in the room where emission was lower. The odds ratio for subjective symptoms with students in classrooms in Building A was not higher than in Building B where the 2E1H concentrations were low. However, a few students limited to Building A did complain of problems with the nasal passages, throat and lower airways. CONCLUSION: Compounds containing 2-ethyl-1-hexyl moiety are presumably hydrolyzed to emit 2E1H when the backing of carpeting material is in contact with concrete floors. Although no significant difference was observed in symptoms between the student groups in the two buildings, this was possibly due to the small sample size. Measures to prevent 2E1H emission and dose-response relationships in sensitive individuals should be studied further.

Adult↗
PubMed2004

Mimicking protein-protein electron transfer: voltammetry of Pseudomonas aeruginosa azurin and the Thermus thermophilus Cu(A) domain at omega-derivatized self-assembled-monolayer gold electrodes.

Well-defined voltammetric responses of redox proteins with acidic-to-neutral pI values have been obtained on pure alkanethiol as well as on mixed self-assembled-monolayer (SAM) omega-derivatized alkanethiol/gold bead electrodes. Both azurin (P. aeruginosa) (pI = 5.6) and subunit II (Cu(A) domain) of ba(3)-type cytochrome c oxidase (T. thermophilus) (pI = 6.0) exhibit optimal voltammetric responses on 1:1 mixtures of [H(3)C(CH(2))(n)()SH + HO(CH(2))(n)()SH] SAMs. The electron transfer (ET) rate vs distance behavior of azurin and Cu(A) is independent of the omega-derivatized alkanethiol SAM headgroups. Strikingly, only wild-type azurin and mutants containing Trp48 give voltammetric responses: based on modeling, we suggest that electronic coupling with the SAM headgroup (H(3)C- and/or HO-) occurs at the Asn47 side chain carbonyl oxygen and that an Asn47-Cys112 hydrogen bond promotes intramolecular ET to the copper. Inspection of models also indicates that the Cu(A) domain of ba(3)-type cytochrome c oxidase is coupled to the SAM headgroup (H(3)C- and/or HO-) near the main chain carbonyl oxygen of Cys153 and that Phe88 (analogous to Trp143 in subunit II of cytochrome c oxidase from R. sphaeroides) is not involved in the dominant tunneling pathway. Our work suggests that hydrogen bonds from hydroxyl or other proton-donor groups to carbonyl oxygens potentially can facilitate intermolecular ET between physiological redox partners.

Azurin↗
PubMed2004

Molten salts bearing anion receptor.

A series of organoboron molten salts prepared by hydroboration of allyl imidazolium type molten salts with various hydroborating reagents such as monobromoborane dimethyl sulfide complex, 9-borabicyclo[3.3.1]nonane (9-BBN) and mesitylborane, and subsequent anion exchange reaction exhibited selective cation transporting property with ionic conductivity of 7.79 x 10(-5) - 6.25 x 10(-6) S cm(-1) at 323 K.

Journal Article↗
PubMed2004

Elastase inhibition reduced death associated with acid aspiration-induced lung injury in hamsters.

This study examined whether the specific inhibition of neutrophil elastase by sivelestat sodium hydrate (sivelestat) reduced deaths associated with severe acute lung injury after hydrochloric acid (HCl) aspiration in hamsters. Animals that received a single intratracheal instillation of HCl (0.2 N, 200 microL) time-dependently died by occlusion of their trachea with inflammatory exudate. In a time course study, these animals developed severe lung injury, peaking 12 to 24 h after HCl instillation, as indicated by hemorrhage and a massive increase in the protein concentration of bronchoalveolar lavage fluid. These changes were closely correlated with neutrophil elastase activity in bronchoalveolar lavage fluid. Sivelestat (0.01, 0.1 and 1 mg/kg/h), when intravenously infused during the first 48 h post-HCl instillation, dose-dependently reduced death in HCl-instilled hamsters. In a separate experiment, analysis of bronchoalveolar lavage fluid parameters and partial pressure of arterial oxygen (PaO(2)) 8 h post-HCl instillation showed that sivelestat at 1 mg/kg/h, i.v. significantly improved both bronchoalveolar lavage fluid parameters and PaO(2) levels with evidence of the inhibition of neutrophil elastase activity in bronchoalveolar lavage fluid. These results suggest that neutrophil elastase plays a significant role in this type of severe acute lung injury that leads to death by respiratory failure.

Animals↗
PubMed2004

One-dimensional ion transport in self-organized columnar ionic liquids.

New fan-shaped ionic liquids forming columnar liquid crystalline phases have been prepared to obtain one-dimensional ion-transporting materials. The ionic liquids consist of two incompatible parts: an imidazolium-based ionic part as an ion-conducting part and tris(alkyloxy)phenyl parts as insulating parts. Two compounds having octyl and dodecyl chains have been synthesized. Self-assembly of these materials leads to the formation of thermotropic hexagonal columnar liquid crystalline states at room temperature. Anisotropic one-dimensional ionic conductivities have been successfully measured by the cells having comb-shaped gold electrodes. The self-organized columns have been aligned macroscopically in two directions by shearing perpendicular and parallel to the electrodes. The ionic conductivities parallel to the column axis are higher than those perpendicular to the axis. The incorporation of lithium salts in these columnar materials leads to the enhancement of the ionic conductivities and their anisotropy. These materials would be useful for anisotropic transportation of ions at the nanometer level.

Journal Article↗
PubMed2004

Structure and direct electrochemistry of cytochrome P450 from the thermoacidophilic crenarchaeon, Sulfolobus tokodaii strain 7.

Cytochrome P450 from thermoacidophilic crenarchaeon, Sulfolobus tokodaii strain 7 (P450st) has been expressed in Escherichia coli and purified at high homogeneity. P450st was crystallized in an orthorhombic system with the space group P2(1)2(1)2(1) and cell dimensions of a=53.6 A, b=55.1 A, and c=130.9 A, and the structure was determined at a 3.0 A resolution. The final R-factor was 0.194 (Rfree=0.235). Structural comparison with cytochrome P450 from S. solfataricus (CYP119) suggests that the region composed of the F to G helices and the Cl- binding site is responsible for the affinity for a ligand coordinating heme iron. Direct electrochemistry of P450st in a didodecyldimethylammonium bromide (DDAB) film on a plastic formed carbon (PFC) electrode has also been demonstrated. A quasi-reversible redox response has been observed even at elevated temperatures of up to 80 degrees C.

Bacterial Proteins↗
PubMed2003

Synthesis, further biological evaluation and pharmacodynamics of newly discovered inhibitors of TNF-alpha production.

(1S,2R)-2-Acylamino-1-methyl-2-phenylethyl phosphate derivatives 2a, 2b, 3a, and 5a, which are conformationally restricted and metabolically stable analogues of (2R)-2-acylamino-2-phenylethyl phosphate derivatives 1a and 1b, are a new class of inhibitors of TNF-alpha production. More efficient alternative synthesis of a key intermediate, (1R,2S)-1-amino-1-(3-methoxyphenyl)propan-2-ol hydrochloride (9), was achieved using one-step, three-component coupling of 3-methoxyphenyl boronic acid (13), (5S)-2,2,5-trimethyl-1,3-dioxolan-4-ol (14), and amino diphenyl methane (15), [as reported in J. Am. Chem. Soc. 1998, 120, 11798]. Evaluation of the hypotensive activity of these compounds was done to assess one of their side effects. Among the compounds tested, the above-mentioned four compounds (2a, 2b, 3a, and 5a) were identified as inhibitors with both sufficient potency and an acceptable safety margin regarding their hypotensive activity. The pharmacodynamics of these compounds were also investigated. Single-dose pharmacokinetic data for compounds 2a, 2b, 3a, and 5a are displayed. These compounds were estimated to be mainly metabolized by the liver in the species tested based on their in vitro stability in tissue homogenates and plasma. A representative compound, 2a, showed good linearity of its plasma concentration after intravenous injection.

Amino Alcohols↗
PubMed2003

Nanostructured anisotropic ion-conductive films.

A flexible self-standing film with layered nanostructures was obtained by in situ photopolymerization of a new smectic liquid-crystalline monomer containing a tetra(oxyethylene) moiety, which forms a macroscopically oriented complex with lithium salts. The resultant films show two-dimensional ionic conductivity.

Journal Article↗
PubMed2003

[Determination of organotin compounds in polyvinyl chloride toys].

Organotin compounds in polyvinyl chloride toys were determined by GC/MS after ethyl derivatization with sodium tetraethylborate. The samples were 12 balls, 12 soft toys, 10 food toys and 13 face masks for children. Monooctyltin, dioctyltin and trioctyltin compounds were found in all face masks at the levels of 74.8-917 micrograms/g. 474-3,960 micrograms/g and 1.0-213 micrograms/g, respectively. They also were detected in 6 balls, 4 soft toys and 1 food toy. Monomethyltin and dimethyltin compounds were found in 8 face masks at the levels of 40.9-227 micrograms/g and 222-1,450 micrograms/g, respectively. Monobutyltin and dibutyltin (DBT) compounds were found in 1 ball, 3 food toys and 5 face masks. In particular, 1 ball and 4 face masks contained toxic DBT at the levels of 527-999 micrograms/g.

Borates↗
PubMed2003

[Studies on the analytical method for epichlorohydrin from internal can coatings].

An improved migration test was developed for determination of trace amounts of epichlorohydrin from internal can coatings. Eight kinds of sample cans, coated mainly with epoxy resin, for foods and beverages were prepared, and both their bodies and lids were tested for migration as follows. A body was filled with n-pentane and soaked for 2 hours at 25 degrees C. A lid was soaked in n-pentane (2 mL/cm2) for 2 hours at 25 degrees C. The test solution was analyzed by GC-FID and GC/MS using two DB-WAX capillary columns with different inside diameters. The limits of quantitation were 0.05 microgram/mL by GC-FID and 0.02 microgram/mL by GC/MS with selected ion monitoring. Recoveries of spiked epichlorohydrin were 99.9-104.5% at the level of 0.05 microgram/mL and 0.5 microgram/mL, with high precision. In this study, no epichlorohydrin was found to have migrated from any of the bodies and lids.

Chromatography, Gas↗
PubMed2002

Highly potent inhibitors of TNF-alpha production. Part 2: identification of drug candidates.

Metabolic stabilization of the chemical lead 1, which is a structurally novel inhibitor of TNF-alpha production, was accomplished by introducing a (1S)-methyl group into the optically active backbone. As a result, 2, 3 and 4 were identified as drug candidates and evaluated pharmacologically. The analysis of an active conformer was also carried out.

Adjuvants, Immunologic↗
PubMed2002

Discovery of novel phosphonic acid derivatives as new chemical leads for inhibitors of TNF-alpha production.

2-(Acylamino)benzylphosphonic acid 6 derived from an artificial substrate of sphingomyelinase was found to show inhibitory activity of TNF-alpha production. Structural optimization was started with the chemical modification of 6. The discovery of another chemical leads 7, 8, 10 and 16 for the development of structurally new inhibitors of TNF-alpha production is reported.

Animals↗
PubMed2002

Highly potent inhibitors of TNF-alpha production. Part II: metabolic stabilization of a newly found chemical lead and conformational analysis of an active diastereoisomer.

Design and synthesis of metabolically stabilized inhibitors of TNF-alpha production, which could be new drug candidates, are reported. Conformational analysis of an active diastereoisomer was performed based on biological evaluations of the conformationally fixed indane derivatives 17 and 18. Structure-activity relationships (SARs) based on biological evaluations of the optically active derivatives are also discussed. Full details including chemistry are reported.

Animals↗
PubMed2002

Highly potent inhibitors of TNF-alpha production. Part I: discovery of new chemical leads and their structure-activity relationships.

Discovery of new chemical leads of inhibitors for TNF-alpha production starting from the chemical modification of 1 is reported. Further biological studies of 1 to disclose the site of its action strongly suggested that 1 inhibits LPS-induced TNF-alpha expression in the liver and spleen of mice. Structure-activity relationships (SARs) are also discussed and full details including the chemistry are reported.

Animals↗