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Biomedical subjects

T Fukui

Publications and source records attributed to T Fukui.

At least 289 records · Page 16Linked to original sources
PubMed1992

Purification and properties of L-histidine decarboxylase from mouse stomach.

L-Histidine decarboxylase was purified to electrophoretic homogeneity from mouse stomach according to a procedure described previously [Ohmori E, Fukui T, Imanishi N, Yatsunami K and Ichikawa A, J Biochem (Tokyo) 107: 834-839, 1990]. The purified enzyme exhibited a specific activity of 750 nmol histamine formed per min per mg protein, which constituted a 37,500-fold purification compared to the crude extract, with a 1.6% yield. The molecular mass of the enzyme was estimated to be 54 kDa by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and 100 kDa by gel filtration. The isoelectric point of the enzyme was determined to be pH 5.4. The Km value for L-histidine was estimated to be 0.29 mM. The single mRNA encoding the amino acid sequence of the mouse stomach enzyme was examined and its size was found to be 2.7 kb. These molecular and catalytic property values of the L-histidine decarboxylase of mouse stomach are quite similar to those of the enzyme from mouse mastocytoma P-815 cells.

Animals↗
PubMed1992

Kinetic resolution of organosilicon compounds by stereoselective dehydrogenation with horse liver alcohol dehydrogenase.

Stereoselective dehydrogenation of three isomers of trimethylsilylpropanol was carried out with horse liver alcohol dehydrogenase (HLADH, EC 1.1.1.1.) and optically active organosilicon compounds were obtained in a water-organic solvent two-layer system with coenzyme regeneration. Furthermore, we examined the effects of the silicon atom on stereoselectivity of HLADH compared to the corresponding carbon compounds. Substitution of the silicon atom for the carbon atom was found to improve the stereoselectivity of HLADH. For example, the optical purity of the remaining 1-trimethylsilyl-2-propanol was higher than 99% enantiomeric excess (ee) at 50% conversion, whereas that of the carbon analogue was 84% ee. This phenomenon was probably ascribable to the bulkiness of the organosilicon compounds derived from their longer Si-C bond. Kinetic analysis in an aqueous monolayer system demonstrated that the specific properties of the silicon atom greatly affected the reactivity of these substrate compounds.

1-Propanol↗
PubMed1992

Bacterial adherence to human gallbladder epithelium.

The adherence of Escherichia coli and Pseudomonas aeruginosa to the epithelium of the gallbladders obtained from 32 patients with negative bile culture was quantified by a scanning electron microscope. Of the gallbladders, 5 were histologically normal (group A), 21 had chronic calculus cholecystitis (group B), and 6 had acute calculus cholecystitis (group C). The data were expressed as the mean +/- S.D. of the numbers of adherent bacteria to 1,000 microns2 of the gallbladder epithelium. The number of adherent E. coli were 0.1 +/- 0.2 in group A, 4.2 +/- 2.8 in group B, and 9.2 +/- 3.3 in group C. A similar result was also observed with P. aeruginosa. The number of adherent bacteria, both of E. coli and P. aeruginosa were significantly higher in group C than in groups A and B, and were also significantly higher in group B compared to group A. The amount of bacterial adherence paralleled that of the degree of epithelial damage, and the normal epithelium proved to have an inhibiting ability. Thus, a secondary bacterial infection is more likely to happen in patients with contaminated bile, and therefore, the treatment for acute cholecystitis should be based either on the results of a bile culture or according to predictive factors for bactibilia.

Acute Disease↗
PubMed1992

Intracellular degradation of poly(3-hydroxybutyrate) granules of Zoogloea ramigera I-16-M.

Intracellular degradation of poly(3-hydroxybutyrate) (PHB) in bacteria is not yet clear. The properties of the autodigestion of native PHB granules from Zoogloea ramigera I-16-M were examined. The release of D(-)-3-hydroxybutyrate was observed only at pH values higher than about 8.5 and at relatively high ionic strength (optimal concentration 200 mM NaCl). Triton X-100 and diisopropylfluorophosphate inhibited this reaction. Addition of the supernatant fraction of Z. ramigera did not increase the release of D(-)-3-hydroxybutyrate from the native PHB granules. On the other hand, using the protease-treated PHB granules from Alcaligenes eutrophus as a substrate, PHB depolymerase activity was detected in the supernatant fraction of Z. ramigera cells. The soluble PHB depolymerase showed similar properties to the enzyme in the PHB granules. Since PHB depolymerase activity was found in fractions containing D(-)-3-hydroxybutyrate oligomer hydrolase activity, which were separated by DEAE-Toyopearl or by Sephacryl S-100, it is possible that the intracellular PHB depolymerase is identical to the oligomer hydrolase which has been purified already.

Biodegradation, Environmental↗
PubMed1992

Angiotensin II stimulates endothelin-1 secretion in cultured rat mesangial cells.

The present study was designed to test two hypotheses: (1) that angiotensin II (Ang II) stimulates endothelin-1 secretion in cultured rat mesangial cells and (2) that atrial and brain natriuretic peptides (ANP and BNP) inhibit the above-mentioned secretion in these cells. Ang II stimulated immunoreactive (ir) endothelin-1 secretion in a concentration-dependent manner between 10(-8) M and 10(-7) M. The protein kinase C (PKC) inhibitors from two chemical classes, H7 and staurosporine, inhibited secretion following such stimulation. The stimulatory effect of Ang II was also abolished in the PKC-depleted cells. Rat ANP(1-28) and rat BNP-45, which are the respective major circulating forms of ANP and BNP in rats, potently inhibited Ang II-stimulated endothelin-1 secretion in a concentration-dependent manner. Inhibition by ANP and BNP of Ang II-stimulated endothelin-1 secretion was paralleled by an increase in the cellular level of cyclic guanosine 5'-monophosphate (GMP). The addition of a cyclic GMP analogue, 8-bromo cyclic GMP, reduced the stimulated endothelin-1 secretion. Rat ANP(5-25) was less effective that rat ANP(1-28) with respect to inhibiting ir-endothelin-1 secretion and increasing cellular cyclic GMP. These findings indicate that Ang II stimulates endothelin-1 secretion in cultured rat mesangial cells by a mechanism probably involving activation of PKC, and that rat ANP and BNP inhibit this stimulated secretion through a cyclic GMP-dependent process.

Angiotensin II↗
PubMed1992

Leucine dehydrogenase from Bacillus stearothermophilus: identification of active-site lysine by modification with pyridoxal phosphate.

We have constructed an efficient expression plasmid for the leucine dehydrogenase gene previously cloned from Bacillus stearothermophilus. The recombinant enzyme was overproduced in Escherichia coli cells to a level of more than 30% of the total soluble protein upon induction with isopropyl beta-D-thiogalactopyranoside. The enzyme could be readily purified to homogeneity by heat treatment and a single step of ion-exchange chromatography. The purified enzyme was inactivated in a time-dependent manner upon incubation with pyridoxal 5'-phosphate (PLP) followed by reduction with sodium borohydride. The inactivation was completely prevented in the copresence of L-leucine and NAD+. Concomitantly with the inactivation, several molecules of PLP were incorporated into each subunit of the hexameric enzyme. Sequence analysis of the fluorescent peptides isolated from a proteolytic digest of the modified protein revealed that Lys80, Lys91, Lys206, and Lys265 were labeled. Among these residues, Lys80 was predominantly labeled and, in the presence of L-leucine and NAD+, was specifically protected from the labeling. Furthermore, a linear relationship of about 1:1 was observed between the extent of inactivation and the amount of PLP incorporated into Lys80. A slightly active mutant enzyme, in which Lys80 is replaced by Ala, was not inactivated at all by incubation with PLP, showing that the inactivation is correlated with the labeling of only Lys80. Lys80is conserved in the corresponding regions of all the amino acid dehydrogenase sequences reported to date. These results suggest that Lys80 is located at the active site and plays an important role in the catalytic function of leucine dehydrogenase.

Amino Acid Oxidoreductases↗
PubMed1992

Structure-activity relationships in human interleukin-1 alpha: identification of key residues for expression of biological activities.

To identify the sites important for the different biological activities of human interleukin-1 alpha (hIL-1 alpha), 56 single-amino acid-substituted mutants of hIL-1 alpha were produced in Escherichia coli using site-directed mutagenesis, and were examined for their biological activities such as mouse lymphocyte activating factor activity (LAF activity), cytostatic activity against human melanoma cells A-375 (A375 activity) and prostaglandin E2 (PGE2) inducing activity in human osteosarcoma cells MG-63 (PEI activity). Two amino acid residues, Asp26 and Asp151, were found to be important for these activities. The replacement of Asp26 by Val caused a decrease in LAF and PEI activities by one or two orders of magnitude and a slight decrease in A375 activity. The Tyr or Phe substitution for Asp151 caused decreases in LAF and A375 activities by one or two orders of magnitude and complete loss of PEI activity. The change from Asp151 to Lys or Arg resulted in marked decrease in LAF activity and complete loss of A375 and PEI activities. Since Asp26 and Asp151 are close to each other in the three-dimensional structure, the region involving these amino acids seems to be important for the biological activities of hIL-1 alpha.

Amino Acid Sequence↗
PubMed1992

Comparing postgraduate medical education at university and non-university hospitals in Japan.

In 1988 the authors surveyed all the teaching hospitals in Japan to evaluate the present status of postgraduate medical education (PGME); they received responses from 67 (84%) of the university and 172 (89%) of the non-university teaching hospitals. It was found that a large proportion of residents had spent two years in a residency without having had a single experience of some of the basic clinical skills. Consequently the residents' confidence in their abilities to perform these skills was low. The residents at the university hospitals, in particular, had had fewer experiences and were less confident about their clinical skills than were the residents at the non-university hospitals. The lack of standard and minimum requirements for PGME in Japan may be the cause of the poor level of acquisition of clinical skills of residents during PGME. Other possible causes are the tendency in Japanese medical society to attach greater importance to academic attainment than to clinical competence and the excessive gravitation of residents toward university hospitals. The authors suggest their results show the necessity to improve the training in basic clinical skills in PGME in Japan, especially in university hospitals.

Clinical Competence↗
PubMed1992

Acute spinal epidural abscess and spinal leptomeningitis: report of 2 cases with comparative neuroradiological and autopsy study.

A 21-year-old male developed back pain, fever, and rapidly progressive quadriparesis. Lumbar tap yielded frank pus which was confirmed on magnetic resonance imaging (MRI) to be located mainly in the cervical epidural space. Conservative antibiotic remedy was partially effective for restoration of the neurological deficits. A 82-year-old female noticed low-back pain which was rapidly accompanied with clouding of consciousness, paraplegia, and sphincter disturbances. Lumbar puncture revealed thick pus which was best depicted on MRI in the thoracolumbar subarachnoid space. At autopsy, spinal subarachnoid abscess or leptomeningitis was confirmed, and a spinal infarction previously unrecognized on MRI was found. Usefulness and shortcomings of MRI in the diagnosis of paraspinal infections are discussed.

Abscess↗
PubMed1992

[An autopsy case of spinal subdural abscess in the aged--comparative study with neuroradiological findings].

An 82-year-old woman without previous medical problem noticed vague back pain on December 31, 1989, and was admitted to a hospital because she developed a fever, a rapidly progressive weakness followed by anesthesia of the lower extremities and sphincter disturbance. On myelography and myelo-CT, the spinal cord appeared to be displaced by an extramedullary mass which partially blocked the subdural space at the level of T-9 to L-1. When transferred to our hospital on January 8, 1990, she was febrile and complaining of headache with meningeal signs. Percussion tenderness was present at T-8 to L-1 spinal spinous process. Neurological examination revealed that the patient had mild consciousness clouding, total paraplegia in the legs, sensory disturbance of a partial degree at L-1 to L-3 and totally below L-3, brisk but equal tendon reflexes in the upper extremities, areflexia in the legs with positive bilateral Babinski signs and sphincter disturbance. Otherwise she was neurologically unremarkable. Acute inflammatory reactions were prominent among the laboratory findings on admission. A lumbar tap yielded purulent fluid with more than 170,000 cells/mm3, 5,000 mg/dl of protein, 44 mg/dl of glucose and culture of the fluid isolated Escherichia coli. T1-weighted sagittal MRI disclosed an ill defined mass which showed the same or locally higher with gadopentetate dimeglumine (Gd-DTPA) signal intensity as soft tissue, compressing the spinal cord anteriorly from T-7 to L-3. The lesion was noticed to have a more extensive rostral-caudal extent than was inferred from myelography and myelo-CT.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗
PubMed1992

[Respiratory regulation system].

In 1874, Kussmaul described "featiful terminal dyspnea" in a case of severe diabetic coma. Probably, this was the first sign of respiratory regulation for metabolic acidosis. After this first case, about 40 years has been needed to establish the theory of acid-base equiribration, namely Henderson-Hasselbalch's equation. pH = 6.1 + log [HCO3-]/[H2CO3] This equation indicates that plasma pH is determined by the ratio of HCO3 concentration and H2CO3 concentration. Because of linear relationship between H2CO3 and PaCO2, pH depend on the ratio of HCO3- and PaCO2. In the state of metabolic acidosis, increase of [H+] stimulates ventilation and decreases the PaCO2. Inversely, in the state of metabolic alkalosis, increase of PaCO2 occurs. These reactions are called "respiratory compensation" or "respiratory regulation". The respiratory regulation system will not retern pH to normal (7.4), but compensation has some limitation which is shown as "SIGNIFICANCE BAND". In this paper, physiological and clinical importance of respiratory regulation and significance band is discussed.

Acid-Base Equilibrium↗
PubMed1992

[Medical decision sciences in the context of clinical pathology].

Medical decision sciences are the scientific approaches to rational decision making under uncertainty. They are arbitrarily subdivided into prescriptive quantitative approach and descriptive psychological approach. The former approach, i.e., decision analysis, is a method of problem solving according to the following procedures: (1) structure the problem (creating a decision tree), (2) apply probabilities, (3) apply values, (4) calculate expected values, (5) perform sensitivity analysis. This methodology makes it clear that once the sensitivity and specificity of a test are determined, estimation of prior probability becomes critically important in interpreting the test results. Usually, prior probability in a given clinical situation is predicted based on patients' demographics, symptoms and previous medical history, physical examination and prior test results. There are, however, many psychological pitfalls leading to wrong estimation of prior probability as follows: (1) premature closure of information processing in the mind, (2) negligence of prior probability from the beginning, (3) regression toward the mean, (4) effect of the number of specimen on the probabilistic fluctuation, (5) gambler's fallacy, (6) effect of vivid memory, (7) selection bias, (8) negligence of negative data, (9) effect of group norm. In order to estimate prior probability as accurately as possible, several strategies are advocated: (1) cognitive psychological approaches, including always thinking about the results of other possible choices, paying attention to the negative data, and trying not to depend on memory as possible, (2) objective estimation using, e.g., the following formula: prior probability = [proportion of patient with abnormal test result-(1-specificity)]/[sensitivity-(1-specificity)].(ABSTRACT TRUNCATED AT 250 WORDS)

Cognition↗
PubMed1992

[Information value of clinical signs and stat tests as indicators of female outpatient urinary tract infection].

This study was designed to evaluate signs and stat tests as an indicator of lower urinary tract infection in female subjects with urogenital complaints at an out-patient clinic. Of various symptoms including hematuria, pollakiuria, dysuria, urinary retention, and micturition pain, pain during micturition was present in 48% of 25 patients with urinary tract infections and in 4% of 27 patients without urinary tract infections, and was the sign with the highest positive predictive value. Comparison and discrimination of the infection and non-infection groups using a single laboratory valuable yielded significant F-statistics for urinary leukocyte esterase (14.5) and leukocyte count in urinary sediment (31.1), and revealed large Mahalanobis' distances for the same variables. Multivariate analysis using a discriminant function of categorical data (Hayashi's Suryoka type 2) revealed that combining occult blood with leukocyte esterase in the urine or combining red cell count with leukocyte count in sediment did not yield substantially smaller misclassification error than did leukocyte esterase alone or leukocyte count alone. It was concluded that neither urinary occult blood nor red cell count in sediment contribute substantially to the prediction of urinary tract infection. For the purpose of detecting urinary tract infection among outpatients, a receiver-operating characteristic analysis demonstrated that the optimal cut-off point in sediment was 3 or more leukocytes per microscopic high power field (x 400). Urinary leukocyte esterase was found to have limitations for use in screening, because its optimal decision level is equivalent to trace esterase reading on the dipstick test.

Ambulatory Care↗
PubMed1992

Holter ECG recordings before and after surgical treatment of hyperthyroidism: circadian pattern of heart rate and arrhythmias.

Holter ECG recordings obtained form 11 hyperthyroid patients (mean age 29.2 +/- 11.4 years) before and after surgical treatment were analyzed regarding heart rate (HR) and arrhythmias. The mean 24-hour HR significantly decreased form 104 +/- 8.8 bpm before to 76.8 +/- 8.9 bpm after the treatment (p less than 0.001), with the most pronounced reduction while asleep form 0:00 AM to 6:00 AM. HR variability also significantly decreased (p less than 0.02), but the circadian rhythm of HR did not change before and after the treatment. Serum T3 level correlated with the mean HR (r = 0.63, p less than 0.05) in hyperthyroid state. The percent changes in T3 (% delta T3) best correlated with the percent changes in the mean HR (% delta mean HR) (r = 0.84, p less than 0.01). The frequencies of supraventricular and ventricular premature contractions significantly decreased after the surgical treatment (p less than 0.01). Normal circadian pattern of hourly HR before the treatment highly suggests that normal adrenergic responsiveness is maintained in hyperthyroidism.

Adult↗
PubMed1991

Potato tuber type H phosphorylase isozyme. Molecular cloning, nucleotide sequence, and expression of a full-length cDNA in Escherichia coli.

Higher plant tissues contain two alpha-glucan phosphorylase isozymes (EC 2.4.1.1), types L and H, localized in the plastid and the cytoplasm, respectively. We already isolated and sequenced a cDNA clone encoding the type L isozyme. Presently, a cDNA clone encoding the type H counterpart was isolated from a cDNA library of immature potato tuber by plaque hybridization, using two oligonucleotide probes synthesized based on the partial amino acid sequences of the type H isozyme. The message encodes a polypeptide of 838 amino acid residues. Sequence comparison of the two potato tuber phosphorylase isozymes revealed two major distinctions; the type L isozyme contains a 78-residue insertion in the middle of the polypeptide chain as well as a 50-residue amino-terminal extension. Except for these extra portions, the two isozyme sequences show an identity of 63%. The entire structural gene for the type H isozyme was inserted 3'-downstream of the strong T7 RNA polymerase promoter in the expression plasmid pET-3b. Escherichia coli BL21 (DE3) cells carrying this plasmid produced active phosphorylase upon induction with isopropyl-beta-D-thiogalactoside at 22 degrees C. The expression is entirely dependent on the temperature; the bacteria did not produce a detectable amount of the active enzyme at 37 degrees C. Addition of pyridoxine to the culture medium was effective for the enzyme production.

Amino Acid Sequence↗