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T Hisano

Publications and source records attributed to T Hisano.

At least 19 recordsLinked to original sources
PubMed1998

Crystal structures of reaction intermediates of L-2-haloacid dehalogenase and implications for the reaction mechanism.

Crystal structures of L-2-haloacid dehalogenase from Pseudomonas sp. YL complexed with monochloroacetate, L-2-chlorobutyrate, L-2-chloro-3-methylbutyrate, or L-2-chloro-4-methylvalerate were determined at 1.83-, 2.0-, 2.2-, and 2.2-A resolutions, respectively, using the complex crystals prepared with the S175A mutant, which are isomorphous with those of the wild-type enzyme. These structures exhibit unique structural features that correspond to those of the reaction intermediates. In each case, the nucleophile Asp-10 is esterified with the dechlorinated moiety of the substrate. The substrate moieties in all but the monochloroacetate intermediate have a D-configuration at the C2 atom. The overall polypeptide fold of each of the intermediates is similar to that of the wild-type enzyme. However, it is clear that the Asp-10-Ser-20 region moves to the active site in all of the intermediates, and the Tyr-91-Asp-102 and Leu-117-Arg-135 regions make conformational changes in all but the monochloroacetate intermediates. Ser-118 is located near the carboxyl group of the substrate moiety; this residue probably serves as a binding residue for the substrate carboxyl group. The hydrophobic pocket, which is primarily composed of the Tyr-12, Gln-42, Leu-45, Phe-60, Lys-151, Asn-177, and Trp-179 side chains, exists around the alkyl group of the substrate moiety. This pocket may play an important role in stabilizing the alkyl group of the substrate moiety through hydrophobic interactions, and may also play a role in determining the stereospecificity of the enzyme. Moreover, a water molecule, which is absent in the substrate-free enzyme, is present in the vicinities of the carboxyl carbon of Asp-10 and the side chains of Asp-180, Asn-177, and Ala-175 in each intermediate. This water molecule may hydrolyze the ester intermediate and its substrate. These findings crystallographically demonstrate that the enzyme reaction proceeds through the formation of an ester intermediate with the enzyme's nucleophile Asp-10.

Binding Sites↗
PubMed1997

Two forms of hemitonic seizure in patients with infantile hemiplegia and epilepsy.

Two forms of unilateral tonic (hemitonic) seizure were observed in two adult epileptic patients with infantile hemiplegia. Their ictal EEGs showed diffuse bilaterally synchronous spike bursts without focal features. In one case, a 31-year-old male, the VTR/EEG recording clearly demonstrated hemitonic seizure which simultaneously involved his paretic limbs and body and were associated with loss of consciousness and autonomic signs. In the other case, a 33-year-old male, partial seizures with secondary unilateral generalization were demonstrated by VTR/EEG recording. The reason why these cases do not exhibit motor seizure activity on the unaffected side despite diffuse bilateral EEG changes, a lower motor threshold in the paretic limbs and inhibitory effect in the brain-stem on undamaged side are postulated. These two cases suggest that hemitonic seizures include heterogeneous characteristics of partial and generalized epilepsy. In patients with diffuse brain damage, such as those with infantile hemiplegia, seizures appear to exhibit complicated patterns, with characteristics of partial and generalized seizures. There is a gradiation of expression of unilateral seizure, range from clearly partial seizures to those with mostly features of generalized seizures, except for an asymmetry of motor expression.

Adult↗
PubMed1996

Increased expression of T-plastin gene in cisplatin-resistant human cancer cells: identification by mRNA differential display.

The cellular resistance to the potent anticancer agent cis-diamminedichloroplatinum(II) (cisplatin) is thought to be mediated by multiple mechanisms. The technique of differential display of mRNAs was applied to various cisplatin-resistant cell lines and the corresponding parental sensitive human bladder, prostatic, and head and neck cancer cells in order to identify genes that underlie cisplatin resistance. Twenty-four clones were confirmed by Northern blot analysis to be expressed differentially between resistant and the corresponding sensitive cells. Partial DNA sequences of the eight clones that showed a threefold or greater increase in expression in either the resistant cells (seven clones) or sensitive cells (one clone) revealed that two were derived from the T-plastin gene and one from the tissue factor gene. The abundance of T-plastin mRNA in cisplatin-resistant T24/DDP10 cell was approximately 12 times that in the parental T24 cells. Transfection of T24/DDP10 cells with a vector encoding full-length T-plastin antisense RNA demonstrated that reduced T-plastin expression was associated with increased sensitivity to cisplatin. These results are consistent with the hypothesis that several mechanisms participate cooperatively in the acquisition of cisplatin resistance in human cancer.

Antineoplastic Agents↗
PubMed1996

Crystal structure of L-2-haloacid dehalogenase from Pseudomonas sp. YL. An alpha/beta hydrolase structure that is different from the alpha/beta hydrolase fold.

L-2-Haloacid dehalogenase catalyzes the hydrolytic dehalogenation of L-2-haloalkanoic acids to yield the corresponding D-2-hydroxyalkanoic acids. The crystal structure of the homodimeric enzyme from Pseudomonas sp. YL has been determined by a multiple isomorphous replacement method and refined at 2.5 A resolution to a crystallographic R-factor of 19.5%. The subunit consists of two structurally distinct domains: the core domain and the subdomain. The core domain has an alpha/beta structure formed by a six-stranded parallel beta-sheet flanked by five alpha-helices. The subdomain inserted into the core domain has a four helix bundle structure providing the greater part of the interface for dimer formation. There is an active site cavity between the domains. An experimentally identified nucleophilic residue, Asp-10, is located on a loop following the amino-terminal beta-strand in the core domain, and other functional residues, Thr-14, Arg-41, Ser-118, Lys-151, Tyr-157, Ser-175, Asn-177, and Asp-180, detected by a site-directed mutagenesis experiment, are arranged around the nucleophile in the active site. Although the enzyme is an alpha/beta-type hydrolase, it does not belong to the alpha/beta hydrolase fold family, from the viewpoint of the topological feature and the position of the nucleophile.

Amino Acid Sequence↗
PubMed1996

Pit structure on bacterial cell surface.

The yellow-pigmented bacterium isolated from a ditch was a gram negative rod with a G+C content of 63 mol%, and was classified in the genus Sphingomonas. Electron microscopy revealed that the bacterial cell surface was covered with many large plaits. When grown in a medium containing a polysaccharide as an essential nutrient, a pit of 0.02-0.1 micrometers in diameter was formed on the cell surface, and a thin section showed the rearrangement of the plaits and the presence of a region where the cell membrane sinks into the cytosol. The dependence of the pit formation on the presence of macromolecule may predict the existence of a direct uptake mechanism for macromolecules through a mouth-like pit, possibly in endocytosis fashion. The confirmation of the pit structure is the first such finding in the history of microbiology and may provide a new insight into the cell morphology and biochemistry of macromolecule transport in microbial cell system.

Alginates↗
PubMed1996

Crystallization and preliminary x-ray crystallographic studies of L-2-haloacid dehalogenase from Pseudomonas sp. YL.

The dimeric L-2-haloacid dehalogenase from Pseudomonas sp. YL, (subunit mass, 26179 Da), has been crystallized by vapor diffusion, supplemented by repetitive seeding, against a 50 mM potassium dihydrogenphosphate solution (pH 4.5) containing 15% (w/v) polyethylene glycol 8,000 and 1% (v/v) n-propanol. The crystals belong to the monoclinic space group C2 with unit cell dimensions of a = 92.21 angstrom, b = 62.78 angstrom, c = 50.84 angstrom, and beta = 122.4 degrees, and contain two dehalogenase dimers in the unit cell. They are of good quality and diffract up to 1.5 angstrom resolution.

Crystallography, X-Ray↗
PubMed1996

Overexpression of multidrug resistance protein gene in human cancer cell lines selected for drug resistance to epipodophyllotoxins.

Overexpression of either the multidrug resistance 1 (MDR1) gene or multidrug resistance protein (MRP) gene is involved in acquisition of multidrug-resistant phenotypes in human cancer cells. In this study we examined whether selection for resistance to the epipodophyllotoxins, etoposide/teniposide (VP16/VM26), could induce overexpression of MDR1 or MRP. We have previously isolated two VP16/VM26-resistant KB cell lines. Two VP16/VM26-resistant KB cell lines, KB/VM-1 and KB/ VM-4, which were selected by stepwise exposure to VM26 had decreased accumulation of [3H]VP16 and increased levels of MRP, but no apparent expression of MDR1 gene was observed. Another VP16/VM26-resistant KB cell line, KB/VP-4, which was further isolated from a VP16-resistant KB cell line, KB/VP-2, had decreased accumulation of [3H]VP16 and showed overexpression of MRP gene, but not that of MDR1 gene. We also isolated a VP16-resistant cell line, IN157/VP-1, from a human glioma cell line IN157. IN157/VP-1 cells showed decreased accumulation of [3H]VP16 and overexpression of MRP gene, but not of MDR1. These findings suggest that selection for resistance to VP16/VM26, preferentially induces overexpression of MRP gene.

ATP-Binding Cassette Transporters↗
PubMed1996

Systemic anaphylaxis after eating storage-mite-contaminated food.

We describe 2 cases in whom systemic anaphylaxis developed shortly after they had eaten food contaminated by a storage mite, Tyrophagus putrescentiae. We were able to demonstrate that these cases were sensitive to the storage mites but not to food allergens, leading us to conclude that the cases' anaphylactic episodes were the result of ingestion of the storage mites. This is the first report of the ingestion of storage mites causing systemic anaphylaxis in sensitive persons.

Adolescent↗
PubMed1994

Re-evaluation of classic senile plaques by three-dimensional analysis.

We evaluated the proportion of classic plaques among all of the senile plaques in four Alzheimer brains (frontal, temporal, occipital and hippocampal areas) by the usual method of two-dimensional analysis using a single methenamine silver-stained section and three-dimensional analysis using a set of serial sections. Three-dimensional analysis showed the number and percentage of classic plaques to be 2-5 times greater than those revealed by two-dimensional analysis. In the hippocampal area of one case, no classic plaques were found by two-dimensional analysis but three-dimensional analysis showed that some classic plaques were present. From these findings, it is suggested that three-dimensional analysis using serial sections is indispensable for subclassifying senile plaques.

Aged↗
PubMed1994

Microtubule reorganization and lysosome redistribution by a viral v-src oncogene, in mouse Balb/3T3 cells expressing human EGF receptor.

The epidermal growth factor (EGF)-induced endocytosis of its receptor is an obligatory pathway for the cellular regulation of the EGF-specific receptor (EGF-R). BNER4 is a mouse Balb/3T3 cell line transfected with human EGF-R complementary DNA (cDNA). B4/src-13 and B4/src-24 are BNER4 cells transfected with a viral oncogene v-src. Indirect immunofluorescence study demonstrated that EGF-R was mostly localized at the perinuclear region in BNER4 cells at 60 min after EGF addition, whereas it was diffusely distributed throughout the cytoplasm in its v-src transfectants. Double indirect immunofluorescence study further confirmed that EGF-R was localized in lysosomes in BNER4 and B4/src-13 cells at 60 min after EGF addition. Intracellular distribution of the Golgi apparatus, clathrin-coated vesicles and early endosomes were similar in all cell lines. However, the lysosomes detected by anti-lysosomal membrane protein (LGP85) antibodies were diffusely distributed throughout the cytoplasm in the v-src transfectacts. By contrast, in the parental BNER4 cells, the lysosomes were mostly localized in the perinuclear region. The organization of microtubules, but not of actin, was markedly different between BNER4 cells and its v-src transfectants. Nocodazole, which depolymerizes microtubules, altered the distribution of the lysosomes and EGF-R in BNER4 cells. Both intracellular lysosome distribution and microtubule organization in nocodazole-treated BNER4 cells were found to be similar to those in its v-src transfectants without nocodazole treatment. These findings support the notion that changes in lysosome distribution may be correlated with microtubule reorganization by v-src in mouse Balb/3T3 cells.

3T3 Cells↗
PubMed1993

Classification of senile plaques by three-dimensional analysis.

We evaluated the number of each senile plaque subtype in Alzheimer's brain by the usual two-dimensional analysis using a single, methenamine silver stained section and three-dimensional analysis using serial section reconstruction. The number of classic plaques was underestimated due to a lack of precision in the usual evaluating procedures (two-dimensional analysis), while that of stellate deposits was overestimated. From these results, it was suggested that the three-dimensional analysis by gapless serial sections was indispensable to examine and subclassify senile plaques.

Alzheimer Disease↗
PubMed1993

Examination of beta-protein deposits in Alzheimer's cerebral cortices by three-dimensional observation.

We examined the location of beta-protein deposits (small diffuse plaques) in the cerebral cortices of Alzheimer's patients and aged nondemented controls by observing the methenamine silver stained serial sections. Some vessels, astrocytes or neuronal elements existed near or within the diffuse plaques, and the preamyloids in the plaques were frequently detected in direct contact with vessels, astrocytes or neuronal elements. Although these findings do not demonstrate that the beta-protein within the senile plaques is produced in various cellular elements of the brain, they suggest some associations between beta-protein deposition and vessels, astrocytes or neuronal elements.

Aged↗
PubMed1993

[Changes in hemodynamics and mixed venous oxygen saturation during intraoperative harvest of autologous platelet-rich plasma].

Intraoperative harvest and transfusion back of platelet-rich plasma (PRP) are an effective procedure to decrease the requirement of blood transfusion. We selected the right atrium as the site for collection and transfusion back of the blood for the harvest of PRP. We studied changes of the hemodynamics and mixed venous oxygen saturation (SvO2 during the intraoperative harvest of autologous PRP in two patients who were undergoing cardiopulmonary bypass (CPB) surgery. The whole blood was collected, processed and the red blood cells were transfused immediately back to the patients using the Plasma Collection System (Haemonetics Model 50). The procedure was accomplished prior to starting CPB. The PRP product was transfused back into the patients at the completion of CPB. The arterial blood pressure, cardiac output, and SvO2 decreased during the collecting phase and immediately returned to the baseline values during the returning phase. No change was observed in the heart rate and arterial oxygen saturation. We conclude that the usage of right atrium as the site for collection and transfusion back of the blood to harvest PRP is effective and useful, and the continuous monitoring of SvO2 is mandatory for detecting the decline of the cardiac output.

Blood Transfusion, Autologous↗
PubMed1993

[Clinical research of patients with epilepsy over age 50 years--a retrospective study of early onset case].

One hundred-sixteen patients with epilepsy over 50 years old were investigated to explore the clinical characteristics of epilepsy in elderly persons. Eighty-four patients having seizures within the last 3 years (residual group) were compared with 32 patients having no seizures within the last 3 years (disappeared group). Regarding the epilepsy type, there were 87 partial epilepsies (64 in the residual group and 23 in the disappeared group) which were approximately 70 percent of the total patients. Generalized epilepsies were comparatively rare in the both groups. Eleven patients with progressive myoclonus epilepsy were observed in the residual group. Regarding the seizure type, generalized convulsive seizures (GCS) were more likely to disappear than partial seizures regardless of number of seizures the patient had. Thirty-eight patients (29 in the residual group and 9 in the disappeared group) had histories of psychiatric problems. Paranoid states were most commonly seen in 12 patients in the residual group and 5 patients in the disappeared group. Intellectual disturbance was seen more frequently in the residual group.

Aged↗
PubMed1992

Characterization of membrane-bound spermidine dehydrogenase of Citrobacter freundii.

Spermidine dehydrogenase found in the membrane fraction of Citrobacter freundii IFO 12681 was solubilized with Triton X-100 and further purified to homogeneity. The properties of the membrane enzyme were almost identical to those obtained from the soluble fraction of the organism with respect to molecular and catalytic properties. Thus, binding properties of the enzyme to the bacterial membrane were checked. The ratio of enzyme activity found in the soluble fraction to the membrane fraction was dependent on salt concentration during cell disruption. A hydrophobic interaction was largely involved in anchoring the enzyme to the membrane fraction. Purified spermidine dehydrogenase from the soluble fraction was readily adsorbed into the membrane fraction in the presence of salt. Spermidine dehydrogenase appeared to be a membrane-bound enzyme localized in the cytoplasmic membranes in a manner that makes a partial release of the enzyme possible during mechanical cell disruption. When spermidine oxidation was done with the resting cells of C. freundii, a stoichiometric formation of two reaction products, 1,3-diaminopropane and gamma-aminobutyraldeyde, was observed without any lag time. These facts indicate that the enzyme is localized on the outer surface of the cytoplasmic membranes or in the periplasmic space of the organism.

Amino Acids↗
PubMed1992

[A patient with gigantic heterotopic gray matter with epileptic seizures].

We report a patient with partial seizure and gigantic heterotopic gray matter. A 23-year-old young man was admitted to our hospital with complaints of frequent epileptic seizures and psychiatric symptoms. There was psychomotor delay in infancy. At the age of 4 years, afebrile convulsions appeared on several occasions. Seizures characterized by a lapse of consciousness started at the age of 13 years. He often stayed in a fantasy world and became very emotional at such time. Cranial CT demonstrated an irregularity in the wall of right lateral ventricle and disappearance of the posterior horn on the same side. This lesion, adjacent to that wall, had a signal intensity that was similar to that of the gray matter on each sequence in MRI. Histopathology of this lesion showed a number of large and small neurons. Therefore, heterotopic gray matter was diagnosed. MRI demonstrated wide cortices suggesting polymicrogyria in the right parietal lobe. Complex partial seizures with eye deviation to the left were recognized. Interictal EEG showed frequent high voltage spikes in the right temporal, fronto-temporal and parieto-occipital areas independently. Therefore, epileptic foci were thought to exist in or around those lesions.

Adult↗