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M Morikawa

Publications and source records attributed to M Morikawa.

At least 91 records · Page 5Linked to original sources
PubMed1999

Molecular diversities of RNases H.

RNase H is an enzyme that specifically cleaves RNA hybridized to DNA. The enzyme is ubiquitously present in various organisms. Single bacterial and eucaryotic cells often contain two RNases H, whereas single archaeal cells contain only one. To determine whether there is a physiological significance in the ubiquity and multiplicity of the enzyme, and whether all enzymes are evolutionarily diverged from a common ancestor, we carried out phylogenetic analyses of the RNase H sequences. In this report, we demonstrated that RNases H are classified into two major families, Type 1 and Type 2 RNases H, of which only the Type 2 enzymes are present in all living organisms, including bacteria, archaea, and eucaryotes, suggesting that they represent an ancestral form of RNases H. Based on this information, we discuss the evolutionary relationships and possible cellular functions of RNases H.

Journal Article↗
PubMed1999

Characterization of petroleum-degrading bacteria from oil-contaminated sites in Vietnam.

Four petroleum-degrading bacterial strains, 2TN-NB, 6TBX-CL, MVK2-5, and XCK, were isolated from various oil-contaminated sites in Vietnam. Determination of the nucleotide sequence of the gene encoding 16S rRNA allowed 2TN-NB to be identified as Acinetobacter sp. and the other three stains as Pseudomonas sp. Among the four isolates, 2TN-NB was most effective in degrading crude oil: in 1 d, it degraded 95% of the crude oil in the culture medium (5%, v/v). The isolated strains could also degrade a sulfur-containing aromatic hydrocarbon, dibenzothiophene (DBT), with low efficiency. Except for MVK2-5, which degraded crude oil least efficiently, the isolates produced biosurfactants in amounts sufficient for structural analysis. FT-IR measurement suggested that strains 6TBX-CL and XCK produced glycolipid-type biosurfactants while that produced by 2TN-NB was of the polysaccharide type.

Journal Article↗
PubMed1999

Identification of the gene encoding esterase, a homolog of hormone-sensitive lipase, from an oil-degrading bacterium, strain HD-1.

The gene encoding an esterase (HDE) was cloned from an oil-degrading bacterium, strain HD-1. HDE is a member of the hormone-sensitive lipase family and composed of 317 amino acid residues with a molecular weight of 33,633. The HDE-encoding gene was expressed in Escherichia coli, and the recombinant protein was purified and characterized. Amino acid sequence analysis indicated that the methionine residue was removed from its NH(2)-terminus. The good agreement of the molecular weights estimated by SDS-PAGE (35,000) and gel filtration (38,000) suggests that it acts in a monomeric form. HDE showed hydrolytic activity towards p-nitrophenyl esters of fatty acids with an acyl chain length of 2 to 14 and tributyrin, whereas it showed little hydrolytic activity towards p-nitrophenyl oleate (C(18)), tricaprylin and triolein. Determination of the kinetic parameters for the hydrolyses of the p-nitrophenyl substrates from C(2) to C(14) indicated that HDE shows a relatively broad substrate specificity. However, comparison of the k(cat)/K(m) values indicated that the C(10)-C(14) substrates are the most preferred ones. Such a preference for substrates with long acyl chains may be a characteristic of HDE.

Amino Acid Sequence↗
PubMed1999

[Presurgical mapping with functional MRI: comparative study with transcranial magnetic stimulation and intraoperative mapping].

PURPOSE: The accuracy of preoperative mappings in patients with brain tumors near the central sulcus using functional magnetic resonance imaging (fMRI) or transcranical magnetic stimulation (TCS) was evaluated by comparative reference to intraoperative mapping. METHODS: The thumb movement was evoked by TCS for the mapping of the motor cortex. After the placement of the marker determined by TCS on the scalp, fMRI under motor tasks consisting of repetitive grasping was performed. For motor cortex activation, an axial oblique plane to maximize gray matter sampling in the rolandic cortex was employed in order to compare these different mapping techniques more precisely. Sixteen patients with brain tumors were included in this study. RESULTS: In nine patients, fMRI disclosed activation in one restricted gyrus or in the localized area around one restricted sulcus. Of these nine patients, preoperative TCS mapping corresponded closely with fMRI in six, while in the remaining three, the TCS marker fell between 1 and 2 cm apart from the fMRI-activated area. However, in these three patients, intraoperative electrocortical stimulation corresponded with the preoperative mapping with fMRI. In six patients, contigucus two gyri were activated by motor tasks. The TCS marker was disclosed on one of the two activated gyri. Of these six patients, the position of the TCS marker and fMRI-activated site corresponded with each other in four cases. They were found on the same gyrus but there was 1.0-2.0 cm distance between them in two cases. Intraoperative somatosensory evoked potential was monitored in two of these six cases. They corresponded well with the mapping by fMRI and TCS together. In only one patient, no significant activation area was obtained by fMRI because of excessive head motion during motor tasks. The TCS marker in this patient was identical with intraoperative electro-cortical stimulation mapping. CONCLUSION: For presurgical planning in patients with brain tumor near the central sulcus, comparative evaluation with fMRI and TCS is applicable and provides accurate functional mapping.

Adult↗
PubMed1999

[CT findings of non-traumatic colorectal perforation].

We retrospectively analyzed CT scans from 23 patients with non-traumatic colorectal perforation. We compared the sensitivity of CT and plain film radiography in the detection of free gas. Free gas was observed in 7 of the 23 cases (30.4%) on plain film radiography and 16 of the 23 cases (69.6%) on CT. Retroperitoneal abscess was demonstrated in 6 of the 7 patients without free gas on CT. Extraluminal air and abscess covered by the omentum and mesenterium were demonstrated in the remaining one patient. The site of perforation was identified in 19 of the 23 patients (82.6%) on CT. CT was useful for demonstrating retroperitoneal free gas, changes in mesenteric fat, extraluminal feces, and tumors. We conclude that CT is indicated in cases of suspected colorectal perforation.

Aged↗
PubMed1998

Stabilization of ribonuclease HI from Thermus thermophilus HB8 by the spontaneous formation of an intramolecular disulfide bond.

To identify factors that contribute to the thermal stability of ribonuclease HI (RNase HI) from Thermus thermophilus HB8, protein variants with a series of carboxyl-terminal truncations and Cys --> Ala mutations were constructed, and their thermal denaturations were analyzed by CD. The results indicate that Cys41 and Cys149 contribute to the protein stability, probably through the formation of a disulfide bond. Peptide mapping analysis for the mutant protein with only two cysteine residues, at positions 41 and 149, indicated that this disulfide bond is partially formed in a protein purified from Escherichia coli in the absence of a reducing reagent but is fully formed in a thermally denatured protein. These results suggest that the thermal stability of T. thermophilus RNase HI, determined in the absence of a reducing reagent, reflects that of an oxidized form of the protein. Comparison of the thermal stabilities and the enzymatic activities of the wild-type and truncated proteins, determined in the presence and absence of a reducing reagent, indicates that the formation of this disulfide bond increases the thermal stability of the protein by 6-7 degreesC in Tm and approximately 3 kcal/mol in DeltaG without seriously affecting the enzymatic activity. Since T. thermophilus RNase HI is present in a reducing environment in cells, this disulfide bond probably is not formed in vivo but is spontaneously formed in vitro in the absence of a reducing reagent.

Amino Acid Sequence↗
PubMed1998

Isolation of novel human cDNA (hGMF-gamma) homologous to Glia Maturation Factor-beta gene.

A novel full-length human cDNA homologous to Glia Maturation Factor-beta (GMF-beta) gene was isolated. Sequence analysis of the entire cDNA revealed an open reading frame of 426 nucleotides with a deduced protein sequence of 142 amino acid residues. The deduced amino acid sequences of its putative product is highly homologous to human GMF-beta (82% identity) and named for GMF-gamma. Northern blot analysis indicated that a message of 0.9 kb long, but not 4.1 kb of GMF-beta, is predominantly expressed in human lung, heart, and placenta.

Amino Acid Sequence↗
PubMed1998

Deaths and injuries due to unexploded ordnance (UXO) in northern Lao PDR (Laos).

Xieng Khousang province in Lao PDR (Laos) is one of the most heavily bombed areas on earth due to the secret bombing during the Vietnam war. This study presents the community-based cumulative incidence of injuries related to unexploded ordnance (UXO) after the war. The data were collected by one of the UK based non-governmental organizations, Mines Advisory Group (MAG). Three districts in the province, reported to have the largest UXO load, were studied. Nearly half of the injuries and deaths involved children below age 15. The case-fatality rate was not different among children and adults. Males were significantly more likely to die of UXO injuries than females. We believe that UXO is an important public health problem in Laos for the following reasons: (1) 22 years after the end of the war, an average of one injury still occurs every other day (2) there is a high incidence among children below age 15 (3) the case-fatality rate is high (4) most injuries involve multiple fragments which usually require complex surgical and medical management skills.

Adolescent↗
PubMed1998

Intracellular localization of migration inhibitory factor-related protein (MRP) and detection of cell surface MRP binding sites on human leukemia cell lines.

The migration inhibitory factor-related proteins (MRPs) MRP-8 and MRP-14 were detected in differentiated human leukemia cell lines (THP-1 and HL-60) by immunocytochemical analysis. They were induced and colocalized in the cytoplasm and in lesser amounts in the nucleus when THP-1 and HL-60 cells were induced to differentiate by 1alpha,25-dihydroxyvitamin D3 or retinoic acid. In a search for a protein capable of binding MRPs, both MRPs were individually produced in insect cells (Sf21) infected with recombinant baculovirus. The purified recombinant MRPs were electrophoretically and antigenically indistinguishable from the native proteins, and their ability to form the MRP8/14 complex was retained. The presence of MRP binding sites was investigated by a binding assay using recombinant MRPs and specific monoclonal antibodies. MRP binding sites were detected on the cell membrane of the human leukemia cell lines THP-1, Raji, and MOLT-4. HL-60 cells treated with 1alpha, 25-dihydroxyvitamin D3 did not express MRP binding sites on the cell membrane, but a high level of MRPs accumulated in the cells. The occurrence of MRP binding sites on the cell surface of leukemia cell lines of monocyte and lymphocyte origin suggests that MRPs, released from neutrophils under certain conditions, may contribute to the activation and recruitment of effector cells to inflammatory lesions.

Animals↗
PubMed1998

Thermostable glycerol kinase from a hyperthermophilic archaeon: gene cloning and characterization of the recombinant enzyme.

The Pk-glpK gene, which encodes glycerol kinase (GK) from a hyperthermophilic archaeon Pyrococcus kodakaraensis KOD1, was cloned and expressed in Escherichia coli. The amino acid sequence of this enzyme (Pk-GK) deduced from the nucleotide sequence showed 57% identity with that of E. coli GK and 47% identity with that of human GK. Pk-GK, which has a molecular weight of 55902 (497 amino acid residues), was purified from E. coli and characterized. Despite the high sequence similarity, Pk-GK and E. coli GK are greatly divergent in structure and function from each other. Unlike E. coli GK, which exists as a tetramer, Pk-GK exists as a dimer. The preferred divalent cation for Pk-GK is Co2+, instead of Mg2+. The optimum pH and temperature for Pk-GK activity are 8.0 and 80 degrees C, respectively. Pk-GK can utilize other nucleoside triphosphates than ATP as a phosphoryl donor. It is fairly resistant to an allosteric inhibitor of E. coli GK, fructose-1,6-bisphosphate. Determination of the kinetic parameters indicates that the Km value of the enzyme is 15.4 microM for ATP and 111 microM for glycerol and its kcat value is 940 s(-1). The enzyme was shown to be fairly resistant to irreversible heat inactivation and still retained 50% of its enzymatic activity even after heating at 100 degrees C for 30 min. Construction of a model for the three-dimensional structure of the enzyme suggests that the formation of extensive ion-pair networks is responsible for the high stability of this enzyme.

Adenosine Triphosphate↗
PubMed1998

Gene cloning and characterization of recombinant RNase HII from a hyperthermophilic archaeon.

We have cloned the gene encoding RNase HII (RNase HIIPk) from the hyperthermophilic archaeon Pyrococcus kodakaraensis KOD1 by screening of a library for clones that suppressed the temperature-sensitive growth phenotype of an rnh mutant strain of Escherichia coli. This gene was expressed in an rnh mutant strain of E. coli, the recombinant enzyme was purified, and its biochemical properties were compared with those of E. coli RNases HI and HII. RNase HIIPk is composed of 228 amino acid residues (molecular weight, 25,799) and acts as a monomer. Its amino acid sequence showed little similarity to those of enzymes that are members of the RNase HI family of proteins but showed 40, 31, and 25% identities to those of Methanococcus jannaschii, Saccharomyces cerevisiae, and E. coli RNase HII proteins, respectively. The enzymatic activity was determined at 30 degreesC and pH 8.0 by use of an M13 DNA-RNA hybrid as a substrate. Under these conditions, the most preferred metal ions were Co2+ for RNase HIIPk, Mn2+ for E. coli RNase HII, and Mg2+ for E. coli RNase HI. The specific activity of RNase HIIPk determined in the presence of the most preferred metal ion was 6. 8-fold higher than that of E. coli RNase HII and 4.5-fold lower than that of E. coli RNase HI. Like E. coli RNase HI, RNase HIIPk and E. coli RNase HII cleave the RNA strand of an RNA-DNA hybrid endonucleolytically at the P-O3' bond. In addition, these enzymes cleave oligomeric substrates in a similar manner. These results suggest that RNase HIIPk and E. coli RNases HI and HII are structurally and functionally related to one another.

Amino Acid Sequence↗
PubMed1998

Sustained left ventricular diastolic dysfunction after exercise in patients with dilated cardiomyopathy.

OBJECTIVE: To investigate the recovery process of exercise induced diastolic dysfunction in heart failure, using Doppler echocardiographic techniques. DESIGN AND PATIENTS: Transmitral flow velocity profiles and standard noninvasive haemodynamic indices were obtained serially over seven days after symptom limited bicycle exercise tests in 18 patients with dilated cardiomyopathy and eight normal subjects. In three patients with cardiomyopathy we also measured the pulmonary capillary wedge pressure for 24 hours after exercise. RESULTS: The intensity of exercise, as assessed by respiratory gas analysis, was lower in patients with dilated cardiomyopathy than in normal subjects. Despite the higher exercise level, all haemodynamic variables returned to baseline within one hour after exercise in normal subjects. In contrast, patients with dilated cardiomyopathy showed a sustained decrease in the peak early diastolic filling velocity and a sustained increase in the deceleration time of early filling for 24 hours or more after exercise. Because other haemodynamic variables recovered within one hour after exercise even in patients with dilated cardiomyopathy, the postexercise changes in ventricular filling were not explained by changes in loading conditions. CONCLUSIONS: Exercise induced diastolic left ventricular dysfunction of the failing heart persists for 24 hours or more after exercise. The efficacy of exercise training on a daily basis in dilated cardiomyopathy requires further evaluation.

Adult↗
PubMed1998

[A study of the differences between the sexes in epidemiology of urolithiasis].

PURPOSE: Uroithiasis seems to be the dominant disease for male. In the recent Japanese epidemiological report, the sexual ratio was reported 2.3:1. This sexual ratio is almost stable from 1960. We studied epidemiologically based on age and stone components the differences between the sexes. PATIENTS AND METHOD: From 1977 to 1996 the upper urinary tract stones which spontaneous excretion or surgical removal in our hospital were the subjects of study, Stone analysis was carried out infrared spectrometer. The statistical analyses were performed by the chi-square test and Mann-Whitney's U test. Significance was declared for a probability value of less than 0.05. RESULTS: In the epidemiology of our treated patients sexual ratio was 1.83:1. We studied sexual ration in age and stone components. The sexual ratio of the patients, 60 years and over was decreased compared with that of the patients under 60 years of age. We found the tendency that the sexual ratio was decreased at 60 years old and over. In the female urolithiasis patients less than 60 years old the frequency of calcium oxalate contained stone patients was predominantly smaller than male. In the female urolithiasis patients of 60 years and over the stone component ratio was similar to male urolithiasis patients. CONCLUSION: In point of frequency sexual ratio was decreased at 60 years old and over. In the female patients less than 60 years old the frequency of calcium oxalate contained stone was predominantly small. This low frequency of calcium oxalate contained stone in the female less than 60 years old seemed to be a factor of the sex differences of frequency.

Adolescent↗
PubMed1998

Evaluation of athletic mouthguard through vibration test on maxillary teeth of human dry skull.

The athletic mouthguard has been used for the prevention of orofacial injuries in contact sports. In this research, the effectiveness of a mouthguard was evaluated for its protection of the maxillary teeth of a human dry skull through experimental modal analysis. The upper central incisor was excited by an electrodynamic shaker. The frequency response functions of the upper teeth were then recorded on an FFT analyzer in order to identify modal shapes of the maxillary arch with and without a mouthguard. In addition, transient response simulations were carried out and decay rates of transient response waves were compared. As a result, when the mouthguard was in place, the resonance peaks of the frequency response functions were dampened, and the nodes of the modal shapes for the anterior teeth were indefinite in the frequency domain over 1 kHz. In the transient response simulation, the decay rates of transient response waves when the mouthguard was used were significantly higher than those when the mouthguard was not used (p<0.01). The results suggest that the mouthguard relieves the stress concentrated on the anterior teeth in a frontal collision by absorbing and dispersing some of the shock energy, thus quickly stopping the vibration of the maxillary teeth. These findings support the clinical fact that mouthguards prevent orofacial injuries in sporting activities.

Athletic Injuries↗
PubMed1998

[Findings of MR and MR cholangiopancreatography in acute cholecystitis].

We retrospectively reviewed magnetic resonance cholangiopancreatography (MRCP) of 25 patients with acute cholecystitis based on clinical, sonographic and surgical findings. Intramural high signal intensity on MRCP was demonstrated in 22 of the 25 patients (88%), and pericholedochal high signal intensity was observed in 6 of the 25 patients (24%). Pericholecystic or perihepatic fluid was demonstrated in 6 of the 25 patients (24%). Gallbladder stones were identified in all 21 patients (100%) by sonography, in 19 of the 21 (90%) by MRCP and in 11 of 18 patients by CT (CT was not performed in other 3 patients). Common bile duct calculi were detected in all 6 patients (100%) by MRCP, in 2 of the 6 (33%) by sonography, and in 5 of the 6 (50%) by CT with confirmation of surgical finding or endoscopic retrograde cholangiography (ERC). MRCP had a high accuracy in diagnosing acute cholecystitis with the finding of intramural high signal intensity. MRCP is an excellent method to evaluate acute biliary disease and may replace CT and ERC in the preoperative evaluation of acute cholecystitis.

Acute Disease↗
PubMed1997

Human IgGFc binding protein (FcgammaBP) in colonic epithelial cells exhibits mucin-like structure.

Cloning a cDNA for human IgGFc binding protein (FcgammaBP) from human colonic epithelial cells reveals an mRNA and coding region of 17 and 16.2 kilobases, respectively. The predicted amino acid sequence contains 12 occurrences of a 400-amino acid cysteine-rich unit resembling that found in mucin. A motif (CGLCGN) in FcgammaBP is conserved in MUC2 and prepro-von Willebrand factor. The N-terminal 450-amino acid sequences are necessary and sufficient to confer IgG Fc binding activity. FcgammaBP mRNA is expressed only in placenta and colonic epithelial cells. These results suggest that FcgammaBP may play an important role in immune protection and inflammation in the intestines of primates.

Amino Acid Sequence↗
PubMed1997

Characterization of a RecA/RAD51 homologue from the hyperthermophilic archaeon Pyrococcus sp. KOD1.

The Pk-rec gene, encoding a RecA/RAD51 homologue from the hyperthermophilic archaeon Pyrococcussp. KOD1, was expressed in Escherichia coli. The recombinant Pk-REC was purified to homogeneity and was shown to be in a dimeric form. A striking property of the purified recombinant Pk-REC was the unusual DNase activity on both single- and double-stranded DNAs along with the ATPase activity. The reaction product of this DNase activity was mononucleotides. The optimum temperature and pH for the DNase activity were 60 degrees C and 8-8.5, respectively. In addition, the metal ion requirement for DNase activity was different from that for the ATPase activity. The protein exhibited no DNase activity in the presence of Zn2+ion, which was one of the most preferable divalent cations for ATPase activity. Another unique characteristic of the recombinant protein was that the reaction product of ATPase activity was AMP instead of ADP.Pk-REC may represent a common prototype of the RecA family proteins with high RecA-like activity.

Adenosine Triphosphatases↗