Biomedical subjects

Koichi Ohno

Publications and source records attributed to Koichi Ohno.

At least 19 recordsLinked to original sources

PubMed2007

Induction of chemoresistance in a cultured canine cell line by retroviral transduction of the canine multidrug resistance 1 gene.

OBJECTIVE: To induce chemoresistance in a normal canine cell line through the transduction of the canine multidrug resistance 1 gene (mdr1). SAMPLE POPULATION: Madin-Darby canine kidney (MDCK) epithelial cell line. PROCEDURES: The full-length canine mdr1 cDNA clone isolated in our laboratory was inserted into a Moloney murine leukemia virus-based vector to construct the retroviral vector, pLNC-cMDR1. After retroviral transduction of pLNC-cMDR1 into MDCK cells, the expression and function of the P-glycoprotein, a product of mdr1, were assessed by immunoblotting, measurement of rhodamine123 (Rh123) retention, and drug sensitivity assays. RESULTS: P-glycoprotein was strongly expressed in cells transduced with pLNC-cMDR1. This P-glycoprotein was fully functional, as demonstrated by the decreased Rh123 retention and the increased resistance to chemotherapeutic drugs. Measured as 50% inhibitory concentrations, resistance increased 59 times to vincristine and 25 times to doxorubicin in MDCK cells after transduction of pLNC-cMDR1. CONCLUSIONS AND CLINICAL RELEVANCE: Transduction of canine mdr1 is an effective method for inducing chemoresistance in normal canine cells. This system may be applicable to the induction of drug resistance in hematopoietic cells.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed2006

Application of RNA interference for inhibiting the replication of feline immunodeficiency virus in chronically infected cell lines.

RNA interference (RNAi) is a process in which double-stranded RNA induces the post-transcriptional sequence-specific degradation of homologous messenger RNA. The present study was carried out to apply the RNAi technology to inhibit the replication of feline immunodeficiency virus (FIV). Four small interfering RNAs (siRNAs) homologous to the FIV gag gene were synthesized and transfected into a feline fibroblastic cell line chronically infected with FIV (CRFK/FIV). These synthetic siRNAs efficiently inhibited the replication of FIV. Next, we examined the effect of retroviral vector-mediated transfer of FIV-specific short hairpin RNA (shRNA) on the replication of FIV in a feline T-cell line chronically infected with FIV (FL4). The retroviral vector-mediated transfer of FIV-specific shRNA was shown to markedly inhibit the replication of FIV in the FL4 cells. These results provide useful information for the development of RNAi-based gene therapy strategy to control FIV infection.

Animals↗

PubMed2006

Genomic cloning of feline Fas ligand gene and characterization of the transcription regulatory region.

The feline Fas ligand gene was molecularly cloned from a feline genomic library and its genomic organization was determined. The feline Fas ligand gene contained four exons and spanned approximately 10 kb in the genome, and thus had the same structure as the human Fas ligand gene. The promoter region of the feline Fas ligand gene was further characterized by deletion mapping. The region between nucleotides -459 and -172 relative to the ATG codon was essential for the promoter activity when transfected into human and feline lymphoid cells. The characterization of the feline Fas ligand gene in the present study will be useful for further investigations of the regulatory mechanism of feline Fas ligand expression.

Amino Acid Sequence↗

PubMed2006

Anisotropic interaction and stereoreactivity in a chemi-ionization process of OCS by collision with He*(2(3)S) metastable atoms.

Ionic-state-resolved collision energy dependence of Penning ionization cross sections for OCS with He*(2(3)S) metastable atoms was measured in a wide collision energy range from 20 to 350 meV. Anisotropic interaction potential for the OCS-He*(2(3)S) system was obtained by comparison of the experimental data with classical trajectory simulations. It has been found that attractive potential wells around the O and S atoms are clearly different in their directions. Around the O atom, the collinear approach is preferred (the well depth is ca. 90 meV), while the perpendicular approach is favored around the S atom (the well depth is ca. 40 meV). On the basis of the optimized potential energy surface and theoretical simulations, stereo reactivity around the O and S atoms was also investigated. The results were discussed in terms of anisotropy of the potential energy surface and the electron density distribution of molecular orbitals to be ionized.

Journal Article↗

PubMed2006

Photodissociation of Mg+-XCH3 (X=F, Cl, Br, and I) complexes. I. Electronic spectra and dissociation pathways.

Photodissociation spectra of Mg+-XCH3 (X=F, Cl, Br, and I) complexes have been measured in the ultraviolet region (225-415 nm). Several fragment ions with and without charge transfer (CT), Mg+, XCH3+, MgX+, MgCH3+, CH3+, and X+, were formed by evaporation (intermolecular bond dissociation) and intracluster reaction (intramolecular bond dissociation) via excited electronic states. Branching ratios of these ions were found to depend both on absorption bands and on halogen atoms. The ground states of the complexes were calculated to have geometries in which the Mg atom lies next to X atom of methyl halide molecules. Positive charges of the complexes are confirmed to be almost localized on Mg. Observed absorption bands were assigned to the transitions of the Mg+2P-2S atomic line perturbed by interactions with methyl halide molecules. Branching ratios of fragment ions can be partly explained by the stability of fragment ions and neutral counterparts. From the excited state potential energy curves along the Mg-X bond distance, dissociation reaction after CT was concluded to proceed predissociatively; potential curve crossings between the initially excited states and repulsive CT states may have a crucial role in the formation of CH3+, XCH3+, and X+. In particular, XCH3+ ions were formed via repulsive CT states having a character of electron excitation from Xnp to Mg+3s.

Journal Article↗

PubMed2006

Photodissociation of Mg+-XCH3 (X=F, Cl, Br, and I) complexes. II. Fragment angular and energy distributions.

Angular and energy distributions of photofragments from Mg+-XCH3 (X=F, Cl, Br, and I) were deduced from time-of-flight (TOF) profiles measured by rotating the polarization direction of the dissociation laser with respect to ion beam direction. The TOF profiles of ICH3+ and MgI+ fragment ions produced from Mg+-ICH3 complex with 266 and 355 nm photons showed clear but opposite recoil anisotropy to each other. In addition, BrCH3+ formed by a dissociation of the Mg+-BrCH3 complex at a photolysis wavelength of 266 nm also showed an anisotropic distribution in the TOF profile which had the same behavior as the profile of ICH3+. For Mg+-FCH3 complex, CH3+ and MgF+ formed with a 266 nm photon had also spatial anisotropy, in which the TOF profile of MgF+ was almost opposite to that of MgI+. These anisotropic distributions were explained by (1) local excitation on the Mg+ ion, (2) rapid dissociation compared with a rotational period of the parent complex, and (3) geometrical structures of the parent complexes. Anisotropy beta parameter values were determined to be +1.30(ICH3+), -0.50(MgI+), +0.74(BrCH3+), and +0.75(CH3+ and MgF+). This dependence on the halogen atom observed in beta values was qualitatively explained by both the geometrical parameters and classical rotational periods of parent complexes. In the product energy distribution, 46%, 40%, 21%, 16%, and 16% of available energies were found to be transferred into translational energies of ICH3+, MgI+, BrCH3 +, CH3+, and MgF+, respectively. These values were compared with energy distributions estimated by a statistical prior distribution and a nonstatistical impulsive model. For ICH3+ and MgI+, the translational energies determined from the measurement had values between those estimated from statistical and nonstatistical models. On the other hand, the energy partitioning for the product ions of BrCH3+, CH3+, and MgF+ was found to be almost statistical. From these considerations, we concluded that nonstatistical processes were more important in the dissociation of Mg+-ICH3 than in other systems.

Journal Article↗

PubMed2006

Aromaticity of giant polycyclic aromatic hydrocarbons with hollow sites: super ring currents in super-rings.

We present a systematic theoretical study based on semi-empirical, Hartree-Fock (HF), and density functional theory (DFT) models of a series of polycyclic aromatic hydrocarbons (PAHs) that exhibit hollow sites. In this study we focus particularly on the magnetic criteria of aromaticity, namely (1)H NMR and nucleus-independent chemical shifts (NICS), and on their relationships with other electronic properties. The computed shifts and NICS indices indicate that an external magnetic field induces exceptionally strong ring currents in even-layered PAH doughnuts, in particular in the layer directly adjacent to the central hole of double-layered compounds. These exceptionally strong ring currents also correlate with particularly small HOMO-LUMO gaps and electronic excitation energies and to abnormally high polarizabilities, indicating in turn that these compounds have a more pronounced metallic character. Comparison is made with further depictions of aromaticity in these systems and in [18]-[66]annulene rings by employing topological, structural, and energetic criteria.

Journal Article↗

PubMed2006

Global reaction route mapping on potential energy surfaces of formaldehyde, formic acid, and their metal-substituted analogues.

Global reaction route mapping of equilibrium structures, transition structures, and their connections on potential energy surface (PES) has been done for MCHO (M = H, Li, Na, Al, Cu) and HCO2M (M = H, Li). A one-after-another technique based on the scaled hypersphere search method has been successfully applied to exploring unknown chemical structures, transition structures, and reaction pathways for organometallic systems. Upon metal substitution, considerable changes of stable structures, reaction pathways, and relative heights of transition structures have been discovered, though some features are similar among the analogues. Al and Cu atoms were found to behave as very strong scissors to cut the CO double bond in MCHO. Energy profiles of the CO insertion into Li-H and Li-CH3 bonds were found to be very similar, especially around the structures where the Li atom is not directly connected with the methyl group, which indicates little effects of alkyl substitution on the reaction route topology.

Journal Article↗

PubMed2006

Collision-energy-resolved Penning ionization electron spectroscopy of thiazole and benzothiazole: study of ionic states and anisotropic interactions between a metastable He(23S) atom and hetero cyclic compounds.

Anisotropic interactions between a metastable He(2(3)S) atom and aromatic heterocyclic compounds (thiazole and benzothiazole) as well as their electronic structures were studied by means of collision-energy/electron-energy resolved two-dimensional Penning ionization electron spectroscopy combined with ab initio molecular orbital calculations. Different collision-energy dependence of partial ionization cross sections (CEDPICS) were clearly observed for different ionic states depending on anisotropic extents of molecular orbitals from which an electron is removed. It was found that thiazole and benzothiazole most strongly attract a He(2(3)S) atom around the region where the nitrogen lone pair orbital extends. For another heteroatom, sulfur, it is relatively weak, but a certain attractive interaction was found for the directions perpendicular to the molecular plane. Benzothiazole was shown to widely attract a He(2(3)S) atom in the out-of-plane directions, since the benzene moiety showed a deeper potential well than the five-membered ring. Assignments of the ionic states including shake-up states were also discussed from observed CEDPICS and ab initio molecular orbital calculations. In particular, for the satellite bands, a negative collision energy dependence of the band intensity was well supported by a configuration-interaction calculation that assigns the satellite bands to be the ionization from pi orbitals accompanying pi-pi or n-pi excitations.

Benzothiazoles↗

PubMed2006

Conversion pathways between a fullerene and a ring among C(20) clusters by a sphere contracting walk method: Remarkable difference in local potential energy landscapes around the fullerene and the ring.

Conversion pathways from a fullerene to a ring and vice versa among C(20) clusters have been explored. A new technique has been developed for discovering multistep reaction pathways, which can be searched by finding constrained energy minima on spheres whose surfaces are contracting from the starting point to the terminal. Local landscapes that appeared on the pathways were found to be considerably different around the fullerene and the ring, from which one can make a new interpretation for no generation of the C(20) fullerene in laser vaporization of graphite.

Journal Article↗

PubMed2006

Large Raman-scattering activities for the low-frequency modes of substituted benzenes: induced polarizability and stereo-specific ring-substituent interactions.

The large nonresonant Raman-scattering activities of the out-of-plane bending and torsional modes of monosubstituted benzene analogs are studied by low-frequency Raman experiments and B3LYP6-31++G(d,p) calculations. Electronic interactions between the sigma orbitals of the substituent and the pi orbitals of the ring are found to enhance the Raman activities, depending on the substituent and its conformation. In the case of tert-butylbenzene [C6H5C(CH3)3] and trimethylphenylsilane [C6H5Si(CH3)3], three single bonds which are linked to the alpha atom of the substituent have low rotational barriers around the joint bond. Nearly free rotation of the substituents leads to a significant probability for one of the single bonds to occupy a conformation close to the vertical configuration with respect to the ring at room temperature. The resultant sigma-pi electronic interaction gives rise to the large Raman activities. In contrast, those possessing a single bond in a coplanar (or nearly coplanar) configuration at the most stable equilibrium state, i.e., anisole (C6H5OCH3), thioanisole (C6H5SCH3), and N-methylaniline (C6H5NHCH3), display no prominent Raman bands for the low-frequency vibrational modes. In these molecules, the sigma-pi conjugation does not take place due to the orthogonal orientation of the orbitals. Strong conformational dependence of the sigma-pi Raman enhancement is clearly obtained for the metastable vertical conformer of thioanisole, for which Raman activities are one-order magnitude greater than those of the coplanar conformer.

Algorithms↗

PubMed2006

Probing anisotropic interaction potentials of unsaturated hydrocarbons with He*(2 3S) metastable atom: attractive-site preference of sigma-direction in C2H2 and pi-direction in C2H4.

State-resolved collision energy dependence of Penning ionization cross sections of acetylene (C2H2) and ethylene (C2H4) with He*(2 3S) metastable atoms was observed in a wide collision energy range from 20 to 350 meV. A recently developed discharge nozzle source with a liquid N2 circulator was employed for the measurements in the low-energy range from 20 to 80 meV. Based on classical trajectory calculations for the energy dependence of the partial ionization cross sections, anisotropic potential energy surfaces for the present systems were obtained by optimizing ab initio model potentials for the chemically related systems Li+C2H2 and C2H4. In the case of C2H2, the global minimum was found to be located around the H atom along the molecular axis with a well depth of 48 meV (ca. 1.1 kcal/mol). On the other hand, a dominant attractive well with a depth of 62 meV (ca. 1.4 kcal/mol) was found in the piCC electron region of C2H4. These findings were discussed in connection with orbital interactions between molecular orbitals of the target molecules and atomic orbitals of the metastable atom. It is concluded that sigma-type unoccupied molecular orbitals of C2H2 and a piCC-type highest occupied molecular orbital of C2H4 play a significant role for the attractive-site preference of sigma direction in C2H2 and pi direction in C2H4, respectively.

Journal Article↗

PubMed2006

Collision-energy-resolved penning ionization electron spectroscopy of phenylacetylene and diphenylacetylene by collision with He*(2(3)s) metastable atoms.

Penning ionization of phenylacetylene and diphenylacetylene upon collision with metastable He*(2(3)S) atoms was studied by collision-energy-/electron-energy-resolved two-dimensional Penning ionization electron spectroscopy (2D-PIES). On the basis of the collision energy dependence of partial ionization cross-sections (CEDPICS) obtained from 2D-PIES as well as ab initio molecular orbital calculations for the approach of a metastable atom to the target molecule, anisotropy of interaction between the target molecule and He*(2(3)S) was investigated. For the calculations of interaction potential, a Li(2(2)S) atom was used in place of He*(2(3)S) metastable atom because of its well-known interaction behavior with various targets. The results indicate that attractive potentials localize in the pi regions of the phenyl groups as well as in the pi-conjugated regions of the acetylene group. Although similar attractive interactions were also found by the observation of CEDPICS for ionization of all pi MOs localized at the C[triple bond]C bond, the in-plane regions have repulsive potentials. Rotation of the phenyl groups about the C[triple bond]C bond can be observed for diphenylacetylene because of a low torsion barrier. So the examination of measured PIES was performed taking into consideration the change of ionization energies for conjugated molecular orbitals.

Journal Article↗

PubMed2006

Essential thrombocythemia in a dog.

A two-year old male Welsh Corgi was referred for persistent thrombocytosis and occasional seizure. Hematological findings indicated marked thrombocytosis, eosinophilia, basophilia and moderate anemia. Bone marrow examination revealed marked megakaryocytic hyperplasia with morphologic abnormality. A diagnosis of essential thrombocythemia was made and the treatment was initiated with combination chemotherapy and maintained by prednisolone and busulfan. The dog successfully achieved complete remission on 100 days after initial presentation and has been good in health without chemotherapy since then.

Animals↗

PubMed2006

C-reactive protein concentration in canine idiopathic polyarthritis.

To investigate the clinical utility of C-reactive protein (CRP) in idiopathic polyarthritis (IPA), its concentration was measured in dogs with IPA. The CRP concentration was markedly increased in all the IPA dogs at the time of diagnosis and decreased significantly in response to the initial corticosteroid treatment; this indicated that CRP can be used as an index for therapeutic response in IPA cases. Furthermore, at 6 months after the diagnosis, a significant association was observed between the CRP concentration at follow-up (6-13 days after the treatment was started) and the frequency of medication ("no or seldom-medicated (NSM)" groups or "continuing medication (CM)" groups). These results suggest that the initial response of CRP to corticosteroid treatment may be a prognostic factor of canine IPA.

Animals↗

PubMed2006

Prognostic factors associated with survival in dogs with lymphocytic-plasmacytic enteritis.

Prognostic factors associated with survival in dogs with lymphocytic-plasmacytic enteritis (LPE) were investigated through a retrospective study. Using case records, 48 dogs diagnosed with LPE were classified as survivors (n = 32) or non-survivors (n = 16), and the clinical and clinicopathological parameters were reviewed between the 2 groups by using univariate and multivariate prognostic analysis. Compared to the hospital population, non-survivors had an overrepresentation of the Shiba breed. Results of univariate analysis indicated that anorexia, severe weight loss, packed cell volume, and total protein were significantly associated with survival for 6 months after diagnosis. In multivariate analysis, anorexia and hypoproteinemia were significantly associated with survival. Furthermore, initial response to treatment was strongly associated with poor prognosis. Based on these clinical and laboratory parameters such as anorexia, hypoproteinemia and initial response to treatment, it may be possible to predict poor prognosis in canine LPE.

Animals↗

PubMed2005

Probing the shape and stereochemistry of molecular orbitals in locally flexible aromatic chains: a penning ionization electron spectroscopy and Green's function study of the electronic structure of biphenyl.

We report on the results of an exhaustive study of the interplay between the valence electronic structure, the topology and reactivity of orbitals, and the molecular structure of biphenyl by means of Penning ionization electron spectroscopy in the gas phase upon collision with metastable He*(2(3)S) atoms. The measurements are compared with one-particle Green's function calculations of one-electron and shake-up valence ionization spectra employing the third-order algebraic diagrammatic construction scheme [ADC(3)]. Penning ionization intensities are also analyzed by means of the exterior electron-density model and comparison with photoelectron spectra: in contrast with the lines originating from sigma orbitals, ionization lines belonging to the pi-band system have large Penning ionization cross sections due to their greater extent outside the molecular van der Waals surface. The involved chemi-ionization processes are further experimentally investigated using collision-energy-resolved Penning ionization electron spectroscopy. The cross sections of pi-ionization bands exhibit a markedly negative collision-energy dependence and indicate that the interaction potential that prevails between the molecule and the He*(2(3)S) atom is strongly attractive in the pi-orbital region. On the other hand, the partial ionization cross sections pertaining to sigma-ionization channels are characterized by more limited collision-energy dependencies, as a consequence of rather repulsive interactions within the sigma-orbital region. A comparison of ADC(3) simulations with the Penning ionization electron spectra and UV photoelectron spectra measured by Kubota et al. [Chem. Phys. Lett. 1980, 74, 409] on thin films of biphenyl deposited at 170 and 109 K on copper demonstrates that biphenyl molecules lying at the surface of polycrystalline layers adopt predominantly a planar configuration, whereas within an amorphous sample most molecules have twisted structures similar to those prevailing in the gas phase.

Journal Article↗

PubMed2005

Development of a cooled He*(23S) beam source for measurements of state-resolved collision energy dependence of Penning ionization cross sections: Evidence for a stereospecific attractive well around methyl group in CH3CN.

A low-temperature discharge nozzle source with a liquid-N(2) circulator for He*(2(3)S) metastable atoms has been developed in order to obtain the state-resolved collision energy dependence of Penning ionization cross sections in a low collision energy range from 20 to 80 meV. By controlling the discharge condition, we have made it possible to measure the collision energy dependence of partial ionization cross sections (CEDPICS) for a well-studied system of CH(3)CN+He*(2(3)S) in a wide energy range from 20 to 350 meV. The anisotropic interaction potential energy surface for the present system was obtained starting from an ab initio model potential via an optimization procedure based on classical trajectory calculations for the observed CEDPICS. A dominant attractive well depth was found to be 423 meV (ca. 10 kcal/mol) at a distance of 3.20 A from the center of mass of CH(3)CN in the N-atom side along the CCN axis. In addition, a weak attractive well (ca. 0.9 kcal/mol) surrounding the methyl group (-CH(3)) has been found and ascribed to the interaction between an unoccupied molecular orbital of CH(3)CN and 2s atomic orbital of He*(2(3)S).

Journal Article↗