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Biomedical subjects

H Nakai

Publications and source records attributed to H Nakai.

At least 109 records · Page 6Linked to original sources
PubMed1997

Molecular cloning of the cDNA for the catalytic subunit of plant DNA polymerase alpha and its cell-cycle dependent expression.

BACKGROUND: DNA polymerase alpha has been studied in considerable detail in yeast and animals. Genetic and biochemical analyses reveal that this enzyme is composed of a heterotetramer and is necessary for replicon initiation and primer synthesis in lagging strand synthesis. In spite of the fact that modes of DNA replication in plants seem to be similar to those in other eukaryotes, very little is known about the biochemical components that participate in DNA replication of plants, including DNA polymerases. RESULTS: Using a 561-base pair DNA fragment, obtained by polymerase chain reaction amplification from a rice cDNA library as a probe, we isolated and sequenced a cDNA homologous to the cDNA for the catalytic subunit of rice DNA polymerase alpha. The encoded polypeptide has extensive homology with the catalytic subunit of DNA polymerase alpha from several species. Furthermore, when the cDNA was expressed in eukaryotic transcription/translation systems, the protein products showed DNA polymerase activity which was inhibited by a monoclonal antibody specific for DNA polymerase alpha. Using RNA gel blot analysis, we found that the levels of mRNA of the catalytic subunit of this enzyme is regulated during the cell-cycle in plant cells. CONCLUSION: This is the first report which describes the cDNA cloning of plant DNA polymerase. We conclude that the principal features of the DNA polymerase alpha catalytic subunit are conserved in plants.

Amino Acid Sequence↗
PubMed1997

Studies on the "signal" constituents for the evaluation of animal crude drugs. III. Nucleic acid components.

An analytical method for determining the contents of 7 nucleic acid bases, 6 ribonucleosides and 5 deoxyribonucleosides in animal crude drugs were established. The contents in free nucleic acid components (the ice-cold extracts) and in total nucleic acid components (the heated extracts) of 8 crude drugs were quite different from one another, indicating that these content patterns could be used as a "signal" of each animal crude drug. For example, the contents of inosine in Lumbricus, uracil in Cervi Parvum Cornu, and guanine and hypoxanthine in Hippocampus were high. The total contents in each crude drug were about twice or three times as large as the free contents. The main degradation products by the heated extracts were adenine and guanine. These results suggested that the contents of nucleic acid components could be one of the "signal" constituents for evaluation of animal crude drugs.

Animals↗
PubMed1997

Glial cyst in the thalamus with intracystic hemorrhage--case report.

A 19-year-old female presented with an unusual glial cyst of the thalamus that caused development of acute hydrocephalus due to hemorrhage and manifested as headache and fainting attacks. Computed tomography showed a large cystic mass lesion in the left thalamus with intracystic hemorrhage. The cyst was subtotally removed. Microscopic examination revealed mild gliosis with marked hemosiderin deposits. The inner surface of the cystic wall lacked an epithelial lining. The diagnosis was glial cyst. Magnetic resonance (MR) imaging 2 months after surgery showed a residual cyst in the left thalamus. However, after 12 months she was asymptomatic, neurologically intact, and MR imaging showed no regrowth of the cyst. Treatment of glial cyst must provide sufficient communication between the cyst and the cerebral ventricles rather than attempt total removal of the cyst, which may present a considerable challenge.

Adult↗
PubMed1997

Aneurysms located at the horizontal segment of the anterior cerebral artery or the middle cerebral artery.

Aneurysms at the horizontal segment of the middle cerebral artery or anterior cerebral artery are relatively rare. The characteristics of 13 cases were analyzed retrospectively. Six of the 13 cases had multiple aneurysms, nine had aneurysmal rupture, and three of these nine were complicated by intracerebral hematoma. Neck clipping of the aneurysm was performed in 11 cases and four developed new cerebral infarction in the territory of the perforating arteries. Overall mortality and morbidity was 15% and 38%, respectively. The outcome for patients with aneurysms at these sites was evidently poorer than for those with aneurysms at other sites.

Aged↗
PubMed1997

A prospective trial of steroid cessation after renal transplantation in pediatric patients treated with cyclosporine and mizoribine.

We conducted a multi-center prospective study to evaluate the safety and efficacy of steroid withdrawal after renal transplantation in children. In 52 children (51 living-related donor transplants and 1 cadaver donor transplant), immunosuppressive therapy was started with cyclosporine (CyA), mizoribine (MZ), methylprednisolone (MPL) and anti-lymphocyte globulin. Administration of MPL was reduced to alternate days more than 6 months after transplantation, and attempts were made to withdraw it. Acute rejection was noted in 19 patients (36.5%) by 1 month after transplantation. The whole-blood CyA trough level using monoclonal antibody was 175.0+/-17.0 ng/ml in patients who developed acute rejection and 282.0+/-25.3 ng/ml in those who did not show acute rejection (p<0.01). During the 37 attempts at alternate-day MPL administration, clinical acute rejection was observed in only 1 patient and chronic rejection in 3. During 10 attempts to withdraw MPL, acute rejection was noted in 3 patients, but graft function recovered to the pre-rejection level after treatment of the acute rejection. At the last observation, graft function was lost in 3 patients, 22 were receiving MPL on alternate days, and MPL had been withdrawn from 7 for a mean period of 16.7 months. The survival rate of the patients and the grafts was 100% and 94% after an average follow-up period of 4 years. Evaluation of growth showed catch-up growth in all patients during the withdrawal period.

Anti-Inflammatory Agents↗
PubMed1997

Effects of a novel free radical scavenger, MCl-186, on ischemic brain damage in the rat distal middle cerebral artery occlusion model.

We investigated the effects of a free radical scavenger, 3-methyl-1-phenyl-2-pyrazolin-5-one (MCl-186), on infarct areas, neurological deficits and regional cerebral blood flow (rCBF), with use of a rat thrombotic distal middle cerebral artery (dMCA) occlusion model to elucidate its possible therapeutic effects on focal cerebral ischemia. In addition, we have attempted to measure 2-oxo-3-(phenylhydrazono)-butanoic acid (OPB), which is the major oxidation product of MCl-186, in the penumbral cortex of a thrombotic dMCA occlusion model. Postischemic treatment with MCl-186 (3 mg/kg) significantly (P < .05) decreased the size of the cerebral infarcts 1 day after dMCA occlusion. MCl-186 (3 mg/kg) significantly (P < .05) improved the neurological deficits 1 day after dMCA occlusion. On the contrary, MCl-186 had no effect on rCBF 1 day after dMCA occlusion. MCl-186 mainly reacted into OPB by peroxidation in rat brain homogenates. Furthermore, the increase in OPB content in the ischemic penumbral cortex tissue was confirmed after 90 min of MCl-186 perfusion. These results suggest that MCI-186 has a protective effect on brain ischemia by reacting with oxygen radicals and that oxygen radicals are closely related to postischemic brain injury.

Animals↗
PubMed1997

Factor VIII Ise (R2159C) in a patient with mild hemophilia A, an abnormal factor VIII with retention of function but modification of C2 epitopes.

We found a patient with mild hemophilia A who had no detectable factor VIII antigen (FVIII:Ag), as shown by two-site ELISA using inhibitor alloantibodies (TK). We then analyzed A2, A2/B, and C2 antigen of the patient's DDAVP-induced FVIII using several anti-FVIII monoclonal antibodies. Factor VIII activity (FVIII:C) was increased from 12 to 42 U/dl by the administration of DDAVP. The DDAVP-induced increases in the A2 and A2/B antigens were 40 and 36 U/dl, respectively. However, the increase in the C2 antigen was only 7.5 U/dl. SSCP analysis and subsequent sequencing demonstrated an Arg to Cys transition at codon 2159. The anti-FVIII:C titer of monoclonal antibody, NMC-VIII/5 which recognized the C2 domain, against normal plasma was 450 Bethesda U/mg of IgG. However, the titer against DDAVP-treated patient's plasma was only 15 Bethesda U/mg. We also tested DDAVP-induced increase in the FVIII:Ag in another mild hemophilia A patient with the same mutation at Arg2159. Increase in his C2 antigen levels was only 19% of those in the A2 and A2/B antigen. We designate this abnormal FVIII as FVIII Ise. Our results show that a missense mutation at Arg2159 to Cys modifies the antigenicity of the C2 domain.

Adolescent↗
PubMed1996

Measurement of anti-factor IX IgG subclasses in haemophilia B patients who developed inhibitors with episodes of allergic reactions to factor IX concentrates.

We have established a simple enzyme-linked immunosorbent assay (ELISA) for the detection of anti-factor IX IgG subclasses in haemophilia B patients with inhibitors. The assay was performed using immobilized purified factor IX. Specific IgG subclasses were detected by peroxidase-conjugated anti-human IgG1,2,3 and 4. Ten plasma samples from 6 haemophilia B patients with inhibitors ranging from 1.0 to 253 Bethesda Units/ml were analyzed. All samples were positive for IgG4. Six out of 10 samples were positive only for IgG4. Three samples were positive for IgG2. Five of the 6 patients had previously had allergic reactions to factor IX concentrates. Three patients had allergic episodes within the past month. Three samples from these latter patients taken on the day when the allergy had occurred showed positive also for IgG1. In later samples, however, taken at 4 days and 4 weeks respectively from two of these same patients. IgG1 was not detected. In two of the five patients in whom allergic reactions had occurred more than one month previously IgG1 was not detected. The results suggested that allergic reactions in patients with haemophilia B treated with factor IX concentrates were associated with the development of the specific IgG1 subclass of antibody to factor IX.

Adolescent↗
PubMed1996

ClpX protein of Escherichia coli activates bacteriophage Mu transposase in the strand transfer complex for initiation of Mu DNA synthesis.

During transposition bacteriophage Mu transposase (MuA) catalyzes the transfer of a DNA strand at each Mu end to target DNA and then remains tightly bound to the Mu ends. Initiation of Mu DNA replication on the resulting strand transfer complex (STC1) requires specific host replication proteins and host factors from two partially purified enzyme fractions designated Mu replication factors alpha and beta (MRFalpha and beta). Escherichia coli ClpX protein, a molecular chaperone, is a component required for MRFalpha activity, which removes MuA from DNA for the establishment of a Mu replication fork. ClpX protein alters the conformation of DNA-bound MuA and converts STC1 to a less stable form (STC2). One or more additional components of MRFalpha (MRFalpha2) displace MuA from STC2 to form a nucleoprotein complex (STC3), that requires the specific replication proteins and MRFbeta for Mu DNA synthesis. MuA present in STC2 is essential for its conversion to STC3. If MuA is removed from STC2, Mu DNA synthesis no longer requires MRFalpha2, MRFbeta and the specific replication proteins. These results indicate that ClpX protein activates MuA in STC1 so that it can recruit crucial host factors needed to initiate Mu DNA synthesis by specific replication enzymes.

ATPases Associated with Diverse Cellular Activitie↗
PubMed1996

The effects of pH on N-methacryloyl glycine primer on bond strength to acid-etched dentin.

To develop a more effective adhesive primer, it is imperative to understand the adhesion mechanisms of the resin to the demineralized dentin through a dentin primer. When the bonding agent was directly applied to the dentin etched by 40 wt % phosphoric acid without a primer pretreatment, the bond strength of the resin to the dentin was 5 MPa. Conversely, when the demineralized dentin was pretreated with the N-methacryloyl glycine (NM alpha A) primer solution with a pH value of 1.5, the bond strength increased considerably to 15 MPa. However, the bond strength dropped dramatically from 15 to 3 MPa when the sodium salt of NM alpha A was added, thereby increasing the pH value of the NM alpha A primer solution from 3.2 to 5.0. When the pH value was increased above 3.5 (pKa value), the number of ionized NM alpha A species increased in the solution. As a result, the bond strength of the resin fell to approximately 3 MPa. This result was obtained despite the 5-micron-thick hybrid layer that was created in the subsurface of the intertubular dentin. The number of unionized NM alpha A species increased by lowering the pH value below 3.5. As a result, the NM alpha A primer provided a higher bond strength of the resin to the demineralized dentin. In contrast, when 10 wt % citric acid containing 3 wt % ferric chloride was applied to the dentin, maximum bond strength was obtained when the pH value of the NM alpha A primer solution was 3.5. The pH dependency of the bond strength obtained following 10 wt % citric acid containing 3 wt % ferric chloride etching is different from the results obtained from 40 wt % phosphoric acid etching. This can be attributed to the difference in the characteristics of the demineralized collageous layer.

Acid Etching, Dental↗
PubMed1996

Non-peptidic inhibitors of human neutrophil elastase: the design and synthesis of sulfonanilide-containing inhibitors.

A novel series of pivaloyloxy benzene derivatives has been identified as potent and selective human neutrophil elastase (HNE) inhibitors. Convergent syntheses were developed in order to identify the inhibitors which are intravenously effective in an animal model. A compound of particular interest is the sulfonanilide-containing analogues. Structure-activity relationships are discussed. Structural requirements for metabolic stabilization are also discussed.

Aniline Compounds↗
PubMed1996

Increased striatal glutamate transporter by repeated intermittent administration of methamphetamine.

We have examined the immunoreactivities of a glutamate (Glu) transporter, GLT-1, in rat brains treated with a single or repeated intermittent administration of methamphetamine (MAP). In the stratum of behaviorally sensitized rate induced by repeated intermittent MAP treatment, GLT-1 immunoreactivities were increased by 51%. There was no difference in the GLT-1 immunoreactivities in all regions examined between rate treated with a single administration of MAP and the control rats. These results suggest that hyperglutamatergic activity in the striatum is involved in the induction of behavioral sensitization caused by MAP.

ATP-Binding Cassette Transporters↗
PubMed1996

Comparison of proliferative activities and metastases between two subtypes classified at the deeply infiltrating sites of colorectal moderately differentiated adenocarcinomas.

Twenty-eight moderately differentiated adenocarcinomas invading beyond the muscularis propria of the colorectum were subclassified as 13 moderate- and 15 poor-subtype tumors based on the histology at the deeply infiltrating sites. Moderately differentiated cancer cells were correlated with liver metastasis and p53 immunoreactivity. Poorly differentiated cancer cells were correlated with lymph node metastases but not to p53 immunoreactivity. The proliferative cell nuclear antigen (PCNA) labeling index (LI), Ki-67 LI and agyrophilic nuclear organizer regions (AgNOR) values determined for the poorly differentiated cancer cells in the poor-subtype tumors were significantly lower than those of moderately differentiated cancer cells in the moderate-subtype tumors. In cells from tumors classified as poor-subtype, poor differentiation was associated with decreased PCNA LI levels, but with unchanged Ki-67 LI and AgNOR values. These results indicate that colorectal adenocarcinoma cells that are histologically subclassified as moderately differentiated have different proliferative and metastatic activities from cancer cells that are poorly differentiated. Moderately differentiated cancer cells are associated with hematogenous metastasis to liver and high proliferative activity, and loss of tubular formation of cancer cells may be fundamentally related to lymph node metastasis and infiltrative growth.

Adenocarcinoma↗
PubMed1996

A case of West syndrome with atypical massive gray matter heterotopia that is well controlled by ACTH therapy.

A 4 month old female infant with atypical asymmetrical massive gray matter heterotopia diagnosed as West syndrome is described. Her seizure initially appeared as afebrile general tonic and clonic convulsion and progressed to typical West syndrome consisting of clusters of myoclonic spasms of the extremities, mainly on the left side, accompanied by head and eye deviation to the right side. Electroencephalogram (EEG) presented typical hypsarrhythmia and cranial computerized tomography (CT) and magnetic resonance imaging (MRI) showed massive heterotopic gray matter in the right hemisphere with the same density or intensity as cortical gray matter. Single photon emission computed tomography (SPECT), using N-isopropyl-p-123I-iodoamphetamine (123 I-IMP), demonstrated decreased blood flow in the ectopic lesion. Although clinical response to several anti-epileptic drugs was poor, her seizures were well controlled by relatively low dose adrenocorticotropic hormone (ACTH) therapy of 0.015 mg/kg per day followed by a combination of valproic acid and clonazepam.

Adrenocorticotropic Hormone↗
PubMed1996

Marginal sealability of resin-modified glass ionomers for base/liner: effect of hygroscopic expansion and bond strength.

The present study investigates the effect of storage in water on hygroscopic expansion and shear bond strength to dentin at periods up to 1 week, of the resin-modified glass ionomers for base/liner, and to analyze the effect on the marginal gaps in dentin cavities. For polishing after storage in water for 1 day, the material indicated significantly smaller marginal gaps both in dentin and in Teflon cavities than in those immediately after light-activation. For the results of after storage in water for 1 day, the material indicated significantly greater bond strength than material immediately after light-activation. The improvement of marginal sealability in dentin cavities may be performed not only by hygroscopic expansion during storage in water but also by greater bond strength after the setting reaction which continues to advance during storage in water.

Analysis of Variance↗
PubMed1996

[Intermittent hydronephrosis. A clinical study in 23 pediatric patients].

BACKGROUND: It is difficult to diagnose intermittent hydronephrosis and to decide the indication of surgical intervention. We investigated 23 cases of intermittent hydronephrosis. METHODS: From 1978 to 1995, a total of 23 patients were diagnosed as intermittent hydronephrosis in our institution. We inspected their clinical features and treatment which had been performed to them. RESULTS: Our study comprised 21 boys and 2 girls, whose mean age was 6 year old. Their chief complaint was intermittent flank pain (left: 21, right: 1, bilateral: 1) accompanied with gross hematuria (30%) and vomiting (39%). When they were asymptomatic, an excretory urogram revealed only mild pelvic dilatation without calyceal distension and kinking of ureteropelvic junction. Split renal function study by RI showed no difference between the affected side and the normal side except one case. When pelvic or calyceal enlargement was confirmed on ultrasonography while they were symptomatic, surgery was indicated. Surgery was performed in 17 cases (74%) including dismembered pyeloplasty in 14 cases, resection of aberrant vessel in 1, relocation of lower pole renal vessel in 1 and nephrectomy in 1. In surgical and histological view points, intrinsic stenosis was seen in 10 cases, extrinsic obstruction caused by aberrant vessels was seen in 4 and ureteral polyp was seen in 3 (bilateral polyp in 1 case). CONCLUSION: They had no more symptoms after operation. Of 23 among followed up cases without surgery, we experienced 2 cases unexpectedly advancing irreversible hydronephrotic change after the last attack, 1 case of gradually progressing hydronephrotic change and 1 case of severe renal dysfunction after many attacks. Therefore intermittent hydronephrosis should be followed up carefully.

Child↗
PubMed1996

[Clinical characteristics and genetic background of secondary amyloidosis associated with rheumatoid arthritis in Japanese].

In order to examine the clinical characteristics and genetic background of secondary amyloidosis associated with rheumatoid arthritis, we analyzed clinical features and HLA typing of 85 patients in a multicenter study. Eighty-five patients with secondary amyloidosis associated RA were studied. The diagnosis of secondary amyloidosis were made on histological findings by biopsy or autopsy. The most common biopsy site was gastrointestinal tract (79.5%). Clinical symptom and the frequency at the time of diagnosis were; diarrhea (35 cases), abdominal pain (22 cases) and vomiting and nausea (16 cases). Abnormalities and the frequency in a laboratory test included proteinuria (49 cases), increased serum creatinine (32 cases), anemia (30 cases) and hematuria (15 cases). Twenty-eight patients were dead and 57 patients were alive at the time of the study. The average duration between diagnosis of amyloidosis and death was 19.4 +/- 18.5 (SD) months among the dead patients. The average duration after diagnosis of amyloidosis was 24.2 +/- 19.5 (SD) months in surviving patients. The causes of death were renal failure complicated with heart failure (6 patients), heart failure alone (3 patients) and renal failure alone (2 patients). Fifty-nine patients in the control group who were negative to amyloid deposition on biopsies at more than one site in the gastrointestinal tract, were clinically compared with patients in the amyloidosis group. No difference were noted in the age of RA occurrence and the stage between the two groups. As to the class, however, the number of patients with severe functional disorder (class 3 or severe) was larger in the amyloidosis group. There were no significant difference between the two groups in Lansbury's activity index. On hematology, biochemistry and urinalysis, the incidences of increased white blood cell count, anemia, increased platelet count, increased serum creatinine, hypoproteinemia, hypoalbuminemia, increased IgA, and increased urine and blood BMG were statistically significantly higher in the amyloidosis group than in the control group. HLA-A, -B, -C, and DR-locus antigens were compared in the 53 patients in the amyloidosis group and in the 59 subjects in the control group. There were no significant differences in frequency of HLA-A, and -B antigens between two groups. Frequency of CW7 antigen was significantly decreased in the amyloidosis group (13.2%) than in the control group (39.0%). Frequency of DR1 antigen was decreased in the amyloidosis group (3.8%) than in the control group (22.0%), although the difference was not significant. These findings suggest the possible involvement of genetic factors in the occurrence of amyloidosis. It is suggested that the occurrence of amyloidosis is suppressed by some genes which are linked with CW7 antigen.

Adolescent↗