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Biomedical subjects

K Shibata

Publications and source records attributed to K Shibata.

At least 91 records · Page 5Linked to original sources
PubMed2001

Nitric oxide synthase activity in neutrophils from patients with localized aggressive periodontitis.

BACKGROUND: Localized aggressive periodontitis (LAgP) is associated with neutrophil dysfunction including defective chemotaxis and reduced calcium influx factor activity. Nitric oxide (NO) and its enzyme, nitric oxide synthase (NOS), have been suggested to be involved in chemotaxis. Some reports, however, were unable to detect either NO or NOS in human neutrophils. In this study, we focused on NOS activity in LAgP neutrophils and examined the involvement of NOS in chemotaxis of normal neutrophils and NOS activity in neutrophils from normal subjects and patients with LAgP. METHODS: Neutrophils from 10 normal subjects and 10 LAgP patients were isolated from peripheral venous blood. Membrane associated-NOS (MA-NOS) and soluble NOS (S-NOS) were extracted from cells with or without FMLP stimulation. NOS activity was measured using the radiolabeled L-arginine to L-citrulline conversion assay. RESULTS: N(omega)-nitro-L-arginine methyl ester (L-NAME), an inhibitor of NOS, significantly inhibited FMLP-induced chemotaxis (P<0.01) and dibutyryl cGMP, an activator of cGMP-dependent protein kinase, significantly attenuated the inhibition by L-NAME (P<0.01). Unstimulated and FMLP-stimulated MA-NOS activity in LAgP neutrophils was statistically significantly higher than that in normal neutrophils (P<0.05). S-NOS activity in LAgP neutrophils was higher than that in normal neutrophils. CONCLUSIONS: This study suggests that NOS is present in human neutrophils and may be involved in FMLP-induced chemotaxis in normal neutrophils. NOS activity is increased in LAgP and is negatively correlated to chemotaxis response.

8,11,14-Eicosatrienoic Acid↗
PubMed2001

Removal of polyA tails from full-length cDNA libraries for high-efficiency sequencing.

We have developed a method to overcome sequencing problems caused by the presence of homopolymer stretches, such as polyA/T, in cDNA libraries. PolyA tails are shortened by cleaving before cDNA cloning with type IIS restriction enzymes, such as GsuI, placed next to the oligo-dT used to prime the polyA tails of mRNAs. We constructed four rice Cap-Trapper-selected, full-length normalized cDNA libraries, of which the average residual polyA tail was 4 bases or shorter in most of the clones analyzed Because of the removal of homopolymeric stretches, libraries prepared with this method can be used for direct sequencing and transcriptional sequencing without the slippage observed for libraries prepared with currently available methods, thus improving sequencing accuracy, operations, and throughput.

Animals↗
PubMed2001

Elevation of blood NAD level after moderate exercise in young women and mice.

We previously reported that the blood NAD levels are decreased by severe exercise, and administration of nicotinamide, a precursor of NAD, improves the endurance capacity of mice. In the present study, we determined whether moderate exercise changes the blood NAD levels in humans and mice. College female students exercised moderately with bike-ergometers. The blood NAD levels elevated after moderate exercise. Mice were forced to swim in a running water pool for 5 min as a moderate exercise, 15 min as a strong exercise, and until exhaustion as a severe exercise (average swimming time was 28.7 min). A 5 min swim gave a result similar to that of moderate exercise by human subjects. However, the blood NAD levels decreased after all-out exercise. The changes in whole blood tryptophan (a precursor of pyridine nucleotides) levels were similar to that in NAD. The glucose levels in whole blood and the non-esterified fatty acid levels in serum decreased according to exercising time. These data are the first demonstration of moderate exercise raising the blood NAD levels in human and mice. Elevation of the blood NAD levels may reflect changes in niacin metabolism that occur in response to exercise.

Adult↗
PubMed2001

Increased conversion ratio of tryptophan to niacin by dietary di-n-butylphthalate.

We reported that the growth promoting activity of di-n-butylphthalate (DBP) was observed when rats were fed with a niacin-free and tryptophan-limiting diet (Shibata et al.. 1982. J Nutr Sci Vitaminol 28, 173-177). The present experiment was performed to investigate whether this phenomenon is attributable to the increase in the conversion ratio of tryptophan to niacin. The weaning rats were fed with a 10% (low protein diet) or 20% protein (conventional protein diet) diet with or without adding 1% DBP. The conversion ratio of tryptophan to niacin was significantly higher in the DBP group than in the control group; for 10% casein diets, it increased two-fold and for 20% casein diet, about five-fold. From these results, the previous finding is possibly explained by DBP increasing the conversion ratio of tryptophan to niacin.

Animals↗
PubMed2001

[Establishment of animal model for elucidating the mechanism of intoxication by the poisonous mushroom Clitocybe acromelalga].

Dietary intake of a poisonous mushroom, Clitocybe acromelalga, causes acromelalgia. The symptom continues for over a month. Some papers reported that treatment with nicotinic acid is effective. We have established an animal model to elucidate the mechanism of toxicity of the poisonous mushroom Clitocybe acromelalga. Diet containing Clitocybe acromelalga was fed to niacin-deficient rats for 24 hours (designated as day 0). The food intake decreased to about one-half compared with that of day before, and body weight loss was noted. Although the diet was returned to the control diet on day 1, the food intake did not recover until day 7, and body weight gain was not seen until day 6. A severe symptom resembling acromelalgia in humans started to appear on day 3. This is the first report of an animal model for the intoxication of Clitocybe acromelalga in humans. Since no similar symptom resembling human intoxication was seen in a previous rodent study, the niacin-free/tryptophan-limited diet used in the present study may have contributed to the result.

Animals↗
PubMed2001

[Effect of feeding with a poisonous mushroom Clitocybe acromelalga on the metabolism of tryptophan-niacin in rats].

The poisonous mushroom Clitocybe acromelalga contains clitidine, which resembles nicotinic acid mononucleotide, and 4-amino-pyridine-2,3-dicarboxylic acid, which resembles quinolinic acid. Both are important intermediates in the tryptophan-niacin pathway. Therefore, we investigated the effect of feeding a niacin-free and tryptophan-limited diet containing the toadstool Clitocybe acromelalga on the metabolism of tryptophan to niacin in rats. The toadstool diet was fed to the rats for only one day (this day was designated day 0). Urinary excretion of intermediates in the tryptophan-niacin pathway, such as anthranilic acid, kynurenic acid, xanthurenic acid, 3-hydroxyanthranilic acid, quinolinic acid, nicotinamide, N1-methylnicotinamide, N1-methyl-2-pyridone-5-carboxamide, and N1-methyl-4-pyridone-3-carboxamide, was higher in the toadstool group than in the control on day 0-day 1 and day 1-day 2. The blood levels of tryptophan and NAD on day 1 were also higher in the toadstool group. Accordingly, intake of Clitocybe acromelalga appeared to increase the conversion of tryptophan to niacin.

Animals↗
PubMed2001

Excretion of bikunin and its fragments in the urine of patients with renal stones.

PURPOSE: Bikunin is a Kunitz-type protease inhibitor found in serum and urine. It has been implicated in urinary stone formation. This study was designed to investigate the role of urinary bikunin in stone formation. MATERIALS AND METHODS: Urinary concentrations of bikunin were measured in 18 male formers of urinary stones 28 to 74 years old and in 77 healthy controls, including 39 males and 38 females, without urological abnormality. A sensitive competitive solid phase enzyme immunoassay was established for urinary bikunin. Bikunin was also qualitatively assessed by Western blot analysis. RESULTS: The mean urinary bikunin-to-creatinine ratio plus or minus standard deviation in stone formers was significantly elevated compared with that in healthy male and female controls (52.9 +/- 46.0 microg./mg. creatinine versus 28.0 +/- 30.4 and 26.5 +/- 21.7, p = 0.005 and 0.006, respectively). By Western blot analysis all urine samples contained authentic 40 kDa. bikunin species. However, a significantly higher proportion of patients was found to have aberrant 25 kDa. bikunin species compared with controls (10 of 18 or 55.6% versus 15 of 77 or 19.5%, p = 0.002). Experiments on de-glycosylation with chondroitinase ABC, amino acid sequencing of the aberrant bikunin species and calcium oxalate crystal growth inhibition assay demonstrated that the 25 kDa. bikunin fragment was identical to de-glycosylated bikunin and less inhibitory on calcium oxalate crystal growth. CONCLUSIONS: If urinary bikunin is important in the pathogenesis of urolithiasis, its effect is probably attributable to the concentration and degree of glycosylation.

Adult↗
PubMed2001

[Plasma (1-->3)-beta-D-glucan level in allergic bronchopulmonary aspergillosis].

We measured the plasma (1-->3)-beta-D-glucan (beta-DG) levels in patients with allergic bronchopulmonary aspergillosis (ABPA) and compared their temporal changes with those of serum IgE and eosinophil counts in the peripheral blood. The subjects were three ABPA patients who were newly diagnosed or had had a relapse between May 1998 and April 1999. Before treatment, all three cases showed high plasma beta-DG values, which declined following corticosteroid treatment with or without oral itraconazole. In two cases, the beta-DG values rose again on relapse. beta-DG values showed a tendency to change in parallel with IgE, although they moved in the opposite direction after a relapse in one case. The present findings suggest that the plasma beta-DG level may be a useful follow-up indicator of the infectious aspect of this disease.

Aged↗
PubMed2001

[Brain infarction related to hepatic arterial infusion chemotherapy].

We examined the occurrence of brain infarction with hepatic arterial infusion chemotherapy for liver cancer. One hundred and eighty-one cases of hepatic arterial infusion chemotherapy were carried out for liver cancer patients in 4 hospitals associated with Osaka University 2nd Dept. of Surgery. These included metastatic (n = 103) and primary (n = 78) liver tumors. The medication was mainly 5-FU with/without CDDP and IFN. Catheters were inserted via the left subclavian artery in 106 cases and via the femoral artery in 75 cases. Among these patients, brain infarctions occurred in seven patients. Occlusions were found in the cerebellum (n = 3), thalamus (n = 1), brain stem (n = 1) and TIA (n = 2). All these patients had catheterization from the left subclavian artery. Furthermore, 64 patients of Ikeda Municipal Hospital were examined and analyzed for brain infarction, in order to eliminate the difference between facilities (all patients in Ikeda Municipal Hospital were catheterized via the left subclavian artery). Many more brain infarctions occurred in metastatic liver cancer patients than in primary liver cancer patients. The hemostasis function deteriorated in primary liver cancer patients, and is thought to be involved in the brain infarction. Six of seven cases of brain infarction occurred in vertebral artery supply area. It may be that the occurrence of brain infarction was related to the flow of the blood vessels.

Aged↗
PubMed2001

[Excitatory effect of noradrenaline on rat airway parasympathetic ganglion neurons].

The effects of noradrenaline (NAd) on neurons acutely isolated from airway parasympathetic ganglia of rats were investigated by use of the nystatin perforaded-patch recording mode. Under current-clamp conditions, an application of 10(-6) M NAd onto the ganglion neurons evoked a depolarization which was accompanied by regenerating repetitive action potentials. NAd concentration-dependently induced inward current with decreasing membrane conductance when a ganglion neuron was held at a holding potential of -40 mV. The half-maximum effective concentration (EC50) of NAd was 1.7 x 10(-7) M, and the response was mimicked by an alpha 1-adrenoceptor agonist cirazoline and was inhibited by WB-4101, an alpha 1A-adrenoceptor antagonist. Oxymetazoline, a partial agonist for an alpha 1A-adrenoceptor, also evoked the inward current with an EC50 of 3.5 x 10(-8) M. The maximum current induced by oxymetazoline (10(-6) M) was 44% of that induced by NAd. NAd also inhibited the amplitude of M-current deactivation induced by hyperpolarizing step from a VH of -25 mV to -50 mV with an EC50 of 2.0 x 10(-4) M. These results suggest that NAd directly depolarizes the airway parasympathetic ganglion neurons of rats associated with an inhibition of M-current through the alpha 1A-adrenoceptors.

Action Potentials↗
PubMed2001

Development of "Standards for the Evaluation of Hospital Infection Control Policies and Procedures, the Second Version".

The Japan Society for Quality in Health Care (JSQua) created "Standards for the Evaluation of Hospital Infection Control Policies and Procedures, the First Version" in 1998 and carried out third-party surveys. Through the experience of those surveys, we revised the standards and created a second version in 1999. The surveyors felt that in using the second version of the standards it would be easier to evaluate the quality of hospital care and that these standards would be more widely applied.

Cross Infection↗
PubMed2000

Reversed-phase high-performance liquid chromatography of nicotinic acid mononucleotide for measurement of quinolinate phosphoribosyltransferase.

A system has been developed for the determination of quinolinate phosphoribosyltransferase (QPRT) activity in liver and kidney homogenates using HPLC. A product, nicotinic acid mononucleotide (NaMN), is separated by reversed-phase chromatography (a Tosoh ODS 80TS was used as an analytical column) using a mixture of 10 mM KH2PO4-K2HPO4 buffer (pH 7.0) containing 1.48 g/l tetra-n-butylammonium bromide-acetonitrile (9:1, v/v) as a mobile phase. The flow-rate was 1.0 ml/min, the detection wavelength was 265 nm. The column temperature was maintained at 40 degrees C. Under these conditions, NaMN was eluted at about 8.1 min. Sample preparation was very straightforward. The reaction mixture of QPRT assay was stopped by immersing the tube into a boiling water bath, the resulting supernatant was filtered, and the filtrate was directly injected into a HPLC system. The total HPLC analysis time was approximately 20 min.

Animals↗
PubMed2000

The N-terminal lipopeptide of a 44-kDa membrane-bound lipoprotein of Mycoplasma salivarium is responsible for the expression of intercellular adhesion molecule-1 on the cell surface of normal human gingival fibroblasts.

The activities to induce TNF-alpha production by a monocytic cell line, THP-1, and ICAM-1 expression and IL-6 production by human gingival fibroblasts were detected in plural membrane lipoproteins of Mycoplasma salivarium. Although SDS-PAGE of the lipoproteins digested by proteinase K did not reveal any protein bands with molecular masses higher than approximately10 kDa, these activities were detected in the front of the gel. A lipoprotein with a molecular mass of 44 kDa (Lp44) was purified. Proteinase K did not affect the ICAM-1 expression-inducing activity of Lp44, but lipoprotein lipase abrogated the activity. These results suggested that the proteinase K-resistant and low molecular mass entity, possibly the N-terminal lipid moiety, played a key role in the expression of the activity. The N-terminal lipid moiety of Lp44 was purified from Lp44 digested with proteinase K by HPLC. Judging from the structure of microbial lipopeptides as well as the amino acid sequence and infrared spectrum of Lp44, the structure of the N-terminal lipid moiety of Lp44 was speculated to be S-(2, 3-bisacyloxypropyl)-cysteine-GDPKHPKSFTEWV-. Its analogue, S-(2, 3-bispalmitoyloxypropyl)-cysteine-GDPKHPKSF, was synthesized. The lipopeptide was similar to the N-terminal lipid moiety of Lp44 in the infrared spectrum and the ICAM-1 expression-inducing activity. Thus, this study suggested that the active entity of Lp44 was its N-terminal lipopeptide moiety, the structure of which was very similar to S-(2, 3-bispalmitoyloxypropyl)-cysteine-GDPKHPKSF.

Amino Acid Sequence↗
PubMed2000

Target volume definition for upper abdominal irradiation using CT scans obtained during inhale and exhale phases.

PURPOSE: To evaluate the clinical utility of a treatment-planning technique involving the use of CT images obtained during both the static exhalation phase and static inhalation phase (two-phase planning). METHODS AND MATERIALS: Ten patients with pancreatic or liver tumors underwent CT scanning under static exhale and inhale conditions, after a period of mild ventilation. By setting image positions differently, we were able to treat the two-phase images as one dataset. Each gross tumor volume (GTV) was contoured separately and the mixed GTV was used for the two-phase treatment planning. Treatment plans were constructed to compare the two-phase plans with the plans constructed using static exhalation images. The shift of the center of the GTV and kidneys and the minimum dose of GTV were then calculated. RESULTS: The shift of the GTV ranged from 2.6 to 27. 3 mm and that of the kidneys from 2.2 to 24 mm. In some patients whose treatment was planned using exhalation planning, the minimum dose of GTV at inhalation was less than 90% of the isocenter dose. CONCLUSION: Two-phase planning is a simple technique that can visualize tumor and organ movement simultaneously using CT. It further defines adequate field margins around the tumor and prevents unexpected radiation exposure to critical organs. Routine use of this technique for upper abdominal irradiation is recommended.

Abdominal Neoplasms↗
PubMed2000

Mechanisms for ovariectomy-induced hyperalgesia and its relief by calcitonin: participation of 5-HT1A-like receptor on C-afferent terminals in substantia gelatinosa of the rat spinal cord.

Chronic treatment with calcitonin in osteoporotic patients alleviates the pain associated with this condition by an unknown mechanism. In ovariectomized rats that develop osteoporosis and hyperalgesia, we examined whether a functional change in serotonergic systems in the spinal dorsal horn was involved, using whole-cell recordings from substantia gelatinosa neurons in spinal cord slices and [(3)H]8-hydroxy-2(di-n-propylamino)tetralin ([(3)H]8-OH-DPAT) binding. Hyperalgesia could be attributed to the elimination of presynaptic inhibition by 5-HT of glutamatergic primary C-afferent terminals and an associated decrease in the density of [(3)H]8-OH-DPAT binding sites whose receptors are neither 5-HT(1A)- nor 5-HT(7)-subtype. These changes in serotonergic systems were restored after chronic treatment with calcitonin. Reversal of 5-HT receptor changes by calcitonin treatment may provide an explanation for its analgesic actions in patients.

8-Hydroxy-2-(di-n-propylamino)tetralin↗
PubMed2000

Synthesis of 1,1-linked galactosyl mannosides carrying a thiazine ring as mimetics of sialyl Lewis X antigen: investigation of the effect of carboxyl group orientation on P-selectin inhibition.

This paper describes the synthesis of 1,1-linked galactosyl mannosides as sialyl Lewis X mimetics that contain a spiro-ring to position the carboxylate group in a well-defined orientation. It was found that compound 4 is more active as a P-selectin inhibitor (IC50 = 19 microM) than the parent disaccharide 2, which contains a flexible carboxyl group (IC50 = 193 microM). This result is consistent with that observed in the previous NMR study of sialyl Lewis X bound to P-selectin. The chemistry described here should be useful for the development of selective inhibitors of E-, P-, and L-selectins.

Carbohydrate Sequence↗