Biomedical subjects

Y Kawamura

Publications and source records attributed to Y Kawamura.

At least 163 records · Page 9Linked to original sources

PubMed1997

Cleft palate and decreased brain gamma-aminobutyric acid in mice lacking the 67-kDa isoform of glutamic acid decarboxylase.

In addition to its role as an inhibitory neurotransmitter, gamma-aminobutyric acid (GABA) is presumed to be involved in the development and plasticity of the nervous system. GABA is synthesized by glutamic acid decarboxylase (GAD), but the respective roles of its two isoforms (GAD65 and 67) have not been determined. The selective elimination of each GAD isoform by gene targeting is expected to clarify these issues. Recently we have produced GAD65 -/- mice and demonstrated that lack of GAD65 does not change brain GABA contents or animal behavior, except for a slight increase in susceptibility to seizures. Here we report the production of GAD67 -/- mice. These mice were born at the expected frequency but died of severe cleft palate during the first morning after birth. GAD activities and GABA contents were reduced to 20% and 7%, respectively, in the cerebral cortex of the newborn GAD67 -/- mice. Their brain, however, did not show any discernible defects. Previous pharmacological and genetic investigations have suggested the involvement of GABA in palate formation, but this is the first demonstration of a role for GAD67-derived GABA in the development of nonneural tissue.

Aging↗

PubMed1997

Serratia liquefaciens as a new host superior for overproduction and purification using the N-acetylneuraminate lyase gene of Escherichia coli.

Serratia liquefaciens was screened as a host strain for effective gene expression and easy purification of the target protein. A model gene, N-acetylneuraminate lyase gene (nanA), fused with the promoter region of Escherichia coli lac operon successfully overproduced the protein independently from the inducer. Since S. liquefaciens grew at lower temperature than E. coli and its proteins were more heat sensitive than those of E. coli, simple incubation at 60 degrees C could inactivate most enzymes but the nanA protein. Subsequent column works for purification, then, became simple and rapid.

Bacterial Proteins↗

PubMed1997

Enzymatic assay for chloride ion with chloride-dependent sarcosine oxidase created by site-directed mutagenesis.

Serum chloride ion is routinely assayed in clinical laboratories. We have developed a new enzymatic assay for determining the chloride ion concentration. The method involves the use of a mutant sarcosine oxidase, which was created as desired by site-directed mutagenesis and showed chloride-dependent activity. The enzyme which is reactivated by the chloride ion forms hydrogen peroxide from sarcosine. The production of hydrogen peroxide is measured by the 4-amino-antipyrine peroxidase system. The increase of the reaction rate was proportional to the chloride ion concentration. A lag time of the time course was not observed, and the reaction rate for a blank was not detected. Therefore, a rate assay could be adopted. A standard curve of the assay reagent was linear up to 180 mM chloride ion of the sample. The specificity for the bromide ion was 43% of that of the chloride ion, although it was 0% for other ion species. When serum samples were used, within-day coefficient variations (CVs) and day-to-day CVs were below 1.5%. A good correlation with the comparison assay was observed by using 160 samples of normal and abnormal patient sera. This method can easily and reliably be used for the accurate determination of chloride ion concentration in serum or other samples.

Amino Acid Sequence↗

PubMed1997

Survival of Vi-capsulated and Vi-deleted Salmonella typhi strains in cultured macrophage expressing different levels of CD14 antigen.

We examined the intracellular survival of Vi-capsulated (lipopolysaccharide; (LPS)-masked) and Vi-deleted (LPS-exposed) Salmonella typhi strains inside macrophage cell lines. Growth of LPS-exposed S. typhi was inhibited in both mouse and human macrophage cell lines. However, the LPS-exposed strain survived in a CD14-deficient mouse macrophage cell lines. Wild-type S. typhi strain, which expressed the Vi antigen and masked LPS, survived in the resting human macrophage cell line. When the Vi-capsulated S. typhi entered the cells, the production of tumor necrosis factor-alpha (TNF-alpha) was suppressed. In contrast, S. typhimurium and LPS-exposed S. typhi stimulated the macrophages to produce a high level of TNF-alpha.

Animals↗

PubMed1997

Ménétrier's disease evaluated serially by abdominal ultrasonography.

We report the case of a 3-year-old boy with Ménétrier's disease who presented with prominent anasarca associated with hypoproteinemia, but no proteinuria. An early sonogram of the stomach demonstrated thickening of the gastric wall which was found to resolve gradually on serial sonograms. Consequently, we considered that the submucosal layer of the gastric wall was particularly thickened as a result of Ménétrier's disease. A gastric biopsy was performed 18 days after onset of the disease, and an electron-microscopic examination of the sample disclosed persistent widening of gastric tight junctions by more than 10 nm. The patient made a full recovery on supportive treatment in 3 weeks. Ultrasonography provided us with a potent tool not only in making the diagnosis, but also in following the course of the disease.

Abdomen↗

PubMed1997

Effect of acetazolamide on cerebral blood flow and tympanic temperature in healthy subjects and patients with subacute subarachnoid haemorrhage.

The influence of the increased cerebral blood flow (CBF) induced by acetazolamide on tympanic temperature (Tty) was examined in three healthy male volunteers and in five patients with subacute subarachnoid haemorrhage (SAH). The CBF was estimated by means of stable xenon-enhanced computed tomography before and after the administration of acetazolamide. The Tty was recorded continuously in both ears using thermistor thermometers. In all subjects, CBF increased ranging from 11% to 108% after acetazolamide administration. In all the healthy subjects and in two patients with mild SAH, Tty was higher than the oesophageal temperature (Toes) and Tty decreased bilaterally, ranging from 0.07 to 0.35 degree C as CBF increased. Three patients with severe SAH were febrile, their Toes exceeding Tty and their Tty rose by 0.30 to 0.53 degree C with increased CBF. These observations suggest that Tty follows brain temperature which changes with an increase in CBF in euthermic subjects as well as in febrile subjects.

Acetazolamide↗

PubMed1997

Circulating soluble selectins in Kawasaki disease.

To investigate the significance of circulating adhesion molecules associated with leucocyte-endothelial cell interaction in Kawasaki disease (KD), serum levels of soluble E-, P-, L-selectin (sE-, sP-, sL-selectin), and vascular cell adhesion molecule-1 (VCAM-1) were measured in 16 patients with KD, eight with other febrile diseases, six with Henoch-Schonlein purpura (HSP), and 10 healthy children using an ELISA. Serum sE-selectin levels from patients in the acute phase of KD were significantly higher than from those in other groups (P < 0.01). The levels of sP-selectin in the subacute phase of KD were significantly higher than in other groups (P < 0.01). Serum sL-selectin levels tended to rise in the convalescent phase of KD. There were also significant correlations between sE-selectin levels and C-reactive protein (r = 0.80, P < 0.0001), and between sP-selectin levels and platelet counts (r = 0.57, P < 0.0001) in KD patients. These data indicate that circulating soluble forms of three selectins may have different kinetics during the clinical course of KD, suggesting that they may reflect its inflammatory process.

Child↗

PubMed1997

Alterations of lactate (+lipid) concentration in brain tumors with in vivo hydrogen magnetic resonance spectroscopy during radiotherapy.

RATIONALE AND OBJECTIVES: The authors determine whether assessment of the relative lactate(+lipid) concentration ([r-Lac(+lip)]) obtained from hydrogen magnetic resonance (MR) spectroscopy is useful for predicting the outcome of radiotherapy on brain tumors. METHODS: Fifty-one hydrogen MR spectroscopic studies were performed in eight patients with primary or metastatic brain tumor before and during radiotherapy. The r-Lac(+lip) calculated as the ratio of lactate(+lipid) peak area to total water was compared before and during radiotherapy in each case. The change of tumor volume measured on magnetic resonance images also was compared. RESULTS: The r-Lac(+lip) substantially decreased in the radiosensitive cases (three lymphomas and one brain cancer) but did not decrease, even at the end of therapy, in the radioresistant cases (two brain cancers and two glioblastomas). CONCLUSION: Assessment of r-Lac(+lip) may adjunctively contribute to the early prediction of the radiotherapeutic effect on brain tumors.

Adult↗

PubMed1997

Genetic identification of members of the genus Corynebacterium at genus and species levels with 16S rDNA-targeted probes.

16S rRNA gene-targeted probes were designed for the identification of corynebacteria at the genus and species levels. The genus-specific probe hybridized all clinically important members of the genus Corynebacterium and could distinguish them from other coryneform bacteria and phylogenetically related high G+C% gram-positive bacteria, including Actinomyces, Rhodococcus, Gordona, Nocardia, Streptomyces, Brevibacterium and Mycobacterium. The species-specific probes for C. jeikeium and C. diphtheriae could differentiated these two species from other members of this genus. The probes were used to select corynebacteria among gram-positive clinical isolates which had been tentatively identified as corynebacteria by biochemical tests. We screened 59 strains with the genus-specific probe; 51 strains hybridized to the genus-specific probe, 8 did not. Of the 51 strains that hybridized to the genus-specific probe, 1 hybridized to the C. diphtheriae species probe and 13 hybridized to the C. jeikeium species probe. The 8 strains that did not hybridize to the genus probe were further characterized by analyzing cell wall diaminopimelic acid and partial 16S rRNA sequencing. The results indicated that these strains were distributed in the genera Arthrobacter and Brevibacterium.

Amino Acids↗

PubMed1997

Transient erythrophagocytosis in Diamond-Blackfan anemia.

We report on a 4-month-old Japanese infant girl with Diamond-Blackfan anemia (DBA) as shown by congenital macrocytic pure red cell hypoplasia with marked reduction of erythroid precursors in bone marrow, reticulocytopenia, increased fetal hemoglobin, and elevated adenosine deaminase activity in peripheral blood. She responded poorly to conventional doses of corticosteroids, however, with high-dose corticosteroids she responded with reticulocytosis and an elevation of hemoglobin level above 12 g/dL. Erythrophagocytosis was noted during the tapering period of prednisone when her hemoglobin level declined to 7.6 g/dL and reticulocyte level to 0.4%. At that time, the erythrophagocytosis was noted in about 60% of marrow histiocytes. These findings were not observed prior to or during the high dose prednisone therapy. We speculate that one of the causes of pure red cell aplasia and reticulocytopenia in DBA is mediated by erythrophagocytosis.

Bone Marrow↗

PubMed1997

[Clinical evaluation of a reagent for detection of DNA of Mycobacterium tuberculosis complex using the ligase chain reaction (LCR) method].

We evaluated the clinical efficacy of LCR MTB, a reagent developed by Abbott in the USA, in the full automatic ligase chain reaction (LCR) for detection of DNA of M. tuberculosis complex using a thermostable ligase. Using 458 samples isolated from patients with tuberculosis, LCR was compared with a smear method and with a culture method, and was also compared with two other methods of gene amplification, MTD and Amplicor, using 340 and 200 of the 458 samples, respectively. The LCR method detected M. tuberculosis in 49.8% (228/458) of the samples, and was superior to the smear method (31.9%, 146/458) and the culture method (39.1%, 179/458) in sensitivity. The LCR method was also superior to the MTD and Amplicor methods; sensitivity were 37.9% (129/340) for MTD vs. 47.6% (162/340) for LCR, and 56.5% (113/200) for Amplicor vs. 59.5% (119/200) for LCR. These favorable results and the convenience of the LCR method, which enables rapid detection of target genes with a high degree of sensitivity, strongly suggest that LCR MTB is useful as a reagent for detection of M. tuberculosis using nucleic acid amplification.

Adolescent↗

PubMed1997

Postoperative changes in the brain: MR imaging findings in patients without neoplasms.

PURPOSE: To determine the time course of postoperative changes in the brain by using magnetic resonance (MR) imaging. MATERIALS AND METHODS: Contrast material-enhanced MR images (32 preoperative, 161 postoperative) in 95 patients who underwent temporal lobectomy for treatment of nonneoplastic epilepsy were evaluated for enhancement pattern at the surgical margin and for the presence of dural enhancement, fluid, and air. Images were obtained 9 hours to 5.6 years after surgery. RESULTS: Surgical margin contrast enhancement was present and increased from 20 hours to 29 days after surgery. The predominant early (days 0-5) pattern was thin linear enhancement (16 of 18 [89%] images). The later (days 6-29) pattern was thick linear or nodular (tumorlike appearance) (11 of 18 [61%] images). Surgical margin enhancement was absent after day 30, although few data were collected 1-5 months after surgery. Dural enhancement was seen on 96 of 97 (99%) postoperative MR images. Extraaxial fluid was seen on all 46 images obtained during the first 30 days and was absent by day 51. Intracranial air was present on 31 of 35 (89%) images obtained during the first 4 days but was absent after day 5. CONCLUSION: Knowledge of the evolution of contrast enhancement patterns on postoperative MR images of the brain can help in differentiating benign from neoplastic changes.

Adolescent↗

PubMed1997

Kinetic assay of serum and urine for urea with use of urease and leucine dehydrogenase.

We describe a new kinetic assay for determining urea in serum or urine with use of urease (EC 3.5.1.5) and leucine dehydrogenase (EC 1.4.1.9). The latter enzyme is suitable for the kinetic assay of NH4+ because its Km value for NH4+ at pH 8.75 is large (approximately 500 mmol/L). Interference from endogenous NH4+ in serum or urine is obviated by subtraction of the assayed endogenous NH4+ value in a sample blank. For serum, within-assay CVs (n = 10) were 0.39-0.58%; day-to-day CVs (n = 10) were 1.56-2.30%. In urine, within-assay CVs (n = 10) were 0.86-1.15%. Analytical recovery of urea (0.893-71.4 mmol/L) added to patients' sera (urea 6.14 mmol/L) was 99.2-105.2%. The calibration curve for serum was linear through zero for urea concentrations up to 142.9 mmol/L and for urine up to 714.3 mmol/L. No influences of added ammonium ion, bilirubin, hemoglobin, ascorbic acid, or Intralipid were observed. The regression equations for this method (y) and conventional methods (x = Determiner-LUN for serum assays, Serotec UUR-R for urine) were: y = 1.016x - 0.12 mmol/L (r = 0.999, S(y/x) = 0.34 mmol/L, n = 100) for sera, and y = 1.070x - 12.6 mmol/L (r = 0.998, S(y/x) = 7.41 mmol/L, n = 100) for urine.

Amino Acid Oxidoreductases↗

PubMed1997

[Clinical significance of 123I-MIBG myocardial scintigraphy in patients with hypertrophic cardiomyopathy].

We studied the abnormality of myocardial sympathetic nervous system in patients with hypertrophic cardiomyopathy using 123I-metaiodobenzylguanidine (MIBG) myocardial scintigraphy in comparison with the parameters of other clinical examinations. In 50 patients with HCM, the heart to mediastinum 123I-MIBG uptake ratio (H/M) was significantly low and washout rate (WR) of 123I-MIBG was significantly high respectively compared with normal subjects (n = 8). H/M was negatively correlated with serum norepinephrine level, wall thickness of left ventricle, left ventricular mass index, left ventricular end diastolic pressure respectively, and WR was positively correlated with those parameters respectively. On the other hand, LF/HF calculated by spectral analysis in holter electrocardiogram was positively correlated with H/M, and negatively correlated with WR. In HCM, H/M in patients with subjective symptoms was significantly lower than that without subjective symptoms, and WR in patients with paroxysmal atrial fibrillation was significantly higher than that without paroxysmal atrial fibrillation. This study revealed that H/M and WR reflected the severity and the difference of disease type in HCM. In conclusion, 123I-MIBG contributes to evaluating more details in diagnosis and pathophysiology of HCM.

3-Iodobenzylguanidine↗

PubMed1997

[Dynamic susceptibility contrast MRI with echo planar imaging using 1.0T system: comparison of different EPI sequences].

Dynamic susceptibility contrast MRI has been performed using the gradient echo sequence on conventional MR imagers. On echo planar imaging (EPI) devices, not only gradient-echo EPI but also spin-echo EPI allow for the monitoring of contrast agent-induced changes in susceptibility. The purpose of this study was to evaluate the contribution of each EPI pulse sequence to susceptibility-induced delta R2(*) through the first pass of a bolus of Gd-DTPA. Thirty healthy volunteers were examined with a 1.0T superconducting MRI unit (IMPACT, Siemens) using EPI. For dynamic susceptibility contrast MRI, we used gradient echo EPI (TE = 60), spin echo EPI (TE = 60) and asymmetric spin echo EPI (TE = 88). Regional relative CBV (rrCBV) maps were generated. On the rrCBV map generated with gradient echo EPI, the high-intensity area of the brain surface was more conspicuous than that generated with spin echo EPI. The rrCBV map generated with asymmetric spin echo EPI represented the contrast between those generated with spin echo EPI and gradient echo EPI. The rrCBV rate between gray matter and white matter with gradient echo EPI was significantly higher than that with spin echo EPI. We suggest that the rrCBV map calculated with spin echo EPI tends to reflect capillary blood volume and the rrCBV map calculated with gradient echo EPI tends to reflect total blood volume.

Adult↗

PubMed1997

[Studies on the bottles of mineral water and the foreign plastic like substances].

The containers of mineral water and the foreign plastic substances which were found in the mineral water were investigated. Most of plastic bottles were made of polyethylene terephthalate (PET) and the caps were made of polypropylene (PP), polyethylene (PE) or aluminum. PE liners were attached to some caps. Most of the foreign plastic substances were PET while others were PE, PP, Teflon and rubber. Some bottles had a scratch on the top inside. The origin of most PET fragments was presumed to be scraped off the bottles by the lowering of the injection nozzle during the water filling process. The sources of the other substances were also determined.

Food Contamination↗

PubMed1996

Mice lacking the 65 kDa isoform of glutamic acid decarboxylase (GAD65) maintain normal levels of GAD67 and GABA in their brains but are susceptible to seizures.

The gene encoding of the 65 kDa isoform of the gamma-aminobutyric acid (GABA)-synthesizing enzyme, glutamic acid decarboxylase (GAD), GAD65, was targeted in mice by homologous recombination. Viable GAD65 -/- mice were obtained with the expected mendelian frequency and displayed no gross morphological defects. Despite the complete loss of GAD65 mRNA and protein in a homozygous mutant, there was no difference in GABA content in the brains of GAD65 +/+, +/-, and -/- mice. As for the other 67 kDa isoform (GAD67), the levels of mRNA and protein were largely unchanged by the GAD65 mutation. General behavior, including locomotor activity and performance in the Morris water maze task, appeared normal, but seizures were more easily induced by picrotoxin and pentylenetetrazol: the latencies to seizures induced by picrotoxin were shorter and the dose of pentylenetetrazol required for induction of seizures was lower.

Animals↗

PubMed1996

Block of Na+ channel by bepridil in isolated guinea-pig ventricular myocytes.

The effects of bepridil, a potent antiarrhythmic agent, on the Na+ current (INa) of single guinea-pig ventricular myocytes were studied using the whole-cell patch-clamp technique. Bepridil inhibited INa in a dose-dependent manner without causing any change in the I-V. relationship for INa. Bepridil suppressed INa with Kd values of 342 and 40 microM when cells were clamped to holding potentials of -140 and -90 mV, respectively. 10 microM bepridil shifted the steady-state inactivation curve for INa toward more negative potentials by 7.7 mV (n = 6). Bepridil also produced marked use-dependent block with a rapid onset. Recovery of INa from inactivation was retarded (time constant 290 ms) at a holding potential of -140 mV in the presence of 10 microM bepridil. When the onset of INa block was studied in experiments using a double-pulse protocol, bepridil blocked INa by 11.5% after a 4-ms pre-pulse, but significantly blocked it after pre-pulses longer than 16 ms. These results suggest that: (1) bepridil has a higher affinity for the inactivated state than the resting state of Na+ channel; (2) the drug also produces an open channel block; and (3) the drug shows a lidocaine-like fast kinetic block of Na+ current.

Animals↗