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Biomedical subjects

K Shibata

Publications and source records attributed to K Shibata.

At least 163 records · Page 9Linked to original sources

Partial purification and characterization of the active entity responsible for inducing interleukin-6 production by human gingival fibroblasts from Mycoplasma salivarium cells.

The active entity responsible for inducing interleukin-6 production by human gingival fibroblasts was partially purified by ion-exchange chromatography from the water-soluble fraction of Mycoplasma salivarium cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the final preparation revealed one densely stained band with a molecular weight of 20.6 kilodaltons and two faint bands with molecular weights of 40.5 and 82.5 kilodaltons. The specific activity of the final preparation was 34-fold higher than that of the starting water-soluble fraction. The interleukin-6-inducing activity was destroyed by proteinase K and reduced 70% by lipoprotein lipase and heat treatment, but was not affected by deoxyribonuclease I or endoglucosidase D. The final preparation induced small amounts of tumor necrosis factor-alpha and interleukin-lbeta in a myelomonocytic cell line, THP-1 cells, but did not induce interleukin-6. The ability of Escherichia coli lipopolysaccharide to stimulate human gingival fibroblasts to release interleukin-6 was dependent upon the presence of serum in the assay medium, but that of the final preparation from M. salivarium was not. Thus, we partially purified the protein(s) from M. salivarium which were capable of stimulating human gingival fibroblasts to release interleukin-6 by a mechanism different from that of E. coli lipopolysaccharide.

Bacterial Proteins↗

Detection of Mycoplasma fermentans in saliva sampled from infants, preschool and school children, adolescents and adults by a polymerase chain reaction-based assay.

Attempts were made to detect Mycoplasma fermentans in saliva sampled from 201 subjects (108 males and 93 females) aged from 4 months to 59 years by a polymerase chain reaction-based assay. M. fermentans was detected in saliva from 110 (54.7%) of 201 subjects, and 10 (28.6%) of 35 subjects aged from 4 months to 3 years. Of ten positive subjects, three were aged from 16 to 23 months and five were from 26 to 31 months. The incidence tended to increase with age up to the teens. The incidence was significantly greater in teenagers than in subjects aged from 7 to 12 years, but there was no significant difference in the incidence between the group of teenagers and each of the groups of subjects older than the teenagers. Thus, it was suggested that M. fermentans colonized the mouth at the age of about 16 months up to the age of 19 years.

Adolescent↗

Transcriptional activation of mRNA of intercellular adhesion molecule 1 and induction of its cell surface expression in normal human gingival fibroblasts by Mycoplasma salivarium and Mycoplasma fermentans.

Lipoproteins in the cell membranes of both Mycoplasma salivarium and Mycoplasma fermentans were demonstrated to trigger the transcription of intercellular adhesion molecule-1 mRNA in normal fibroblasts isolated from human gingival tissue and to induce its cell surface expression by a mechanism distinct from that of Escherichia coli lipopolysaccharide. The lipid moiety of the lipoproteins was suggested to play a key role in the expression of the activity.

Bacterial Proteins↗

Expression of adrenomedullin in feto-placental circulation of human normotensive pregnant women and pregnancy-induced hypertensive women.

The adrenomedullin (AM) peptide and the expression of AM messenger RNA (mRNA) from feto-maternal tissues of 22 normotensive pregnant women and from 7 women with pregnancy-induced hypertension (PIH) during third-trimester were examined to clarify the pathophysiological features of PIH. Samples of the placenta, uterine muscle, umbilical artery, and fetal membranes were obtained from each patients under informed consent. The AM peptide was measured by a radioimmunoassay and the AM mRNA level was analyzed by Northern hybridization. The total immunoreactive AM (fmol/mg wet tissue) was significantly increased in the fetal membranes (1.95+/-0.20 vs. 3.03+/-0.44) and the umbilical artery (0.11+/-0.01 vs. 0.15+/-0.02) of the patients with PIH. On the other hand, the AM mRNA level was higher in the umbilical artery, and lower in the fetal membranes in the patients with PIH. The present results thus suggest that the changes in the expression of AM in fetal membrane and umbilical artery in PIH may play an important role in the fetal and maternal circulation.

Adrenomedullin↗

Sustained release liquid preparation using sodium alginate for eradication of Helicobacter pyroli.

We prepared a new liquid preparation for eradication of Helicobacter pylori (HP), and examined drug release in vitro and in vivo. The liquid preparation mainly consisted of a sodium alginate (AG) aqueous solution containing ampicillin (ABPC), an antibiotic drug, or methylene blue, a dye. Drug release was retarded by Ca pre-treatment (0.10 M, 20 s) of the AG preparation in in vitro drug release studies due to gel-formation at the liquid surface. In in vivo experiments, the AG preparations were administered orally to rats. The rats were divided into two groups, with or without pre-administration of ranitidine hydrochloride (RH, an H2-blocker). The total remaining % of ABPC in the stomach was high in the rats administered the AG preparation compared to the ABPC solution. The AG preparation might float in the stomach without adhering to the gastric wall in the rats without pre-administration of RH. The total remaining % of ABPC at 30 min was almost 100% in the RH pre-administration rats administered the AG preparation, and about 80% of the drug existed in fraction 2 (implying adhesion of the preparation on the gastric mucus). At 60 min, the total remaining % in the AG preparation plus Ca (mean 87%) increased about 2-fold compared to that in the AG preparation alone (mean 44%). In this case, a large portion of the remaining ABPC also existed in fraction 2. This preparation may be useful for eradication of HP.

Administration, Oral↗

Preparation and drug retention of biodegradable chitosan gel beads.

Chitosan (CS) gel beads containing drug could be prepared in amino acid solutions of pH about 9, despite the requirement for a pH above 12 for gelation in water. This phenomenon was observed not only in amino acid solutions but also in solutions of compounds having amino groups. A solute concentration of more than 10% was required for preparation of gel beads at pH 9. Gelation of the CS beads required about 25 to 40 min, depending on the species of amino acid. Lidocaine hydrochloride (LC) as a model drug was retained in the beads to about 20 to 35% of the theoretical total amount, despite being a water-soluble drug. The release of LC from the CS gel beads was prolonged. The release pattern was not affected by the species of amino acid or CS, or the preparation time.

Biocompatible Materials↗

Naftopidil, a novel alpha1-adrenoceptor antagonist, displays selective inhibition of canine prostatic pressure and high affinity binding to cloned human alpha1-adrenoceptors.

The pharmacological profiles of the alpha1-adrenoceptor antagonists naftopidil, tamsulosin and prazosin were studied in an anesthetized dog model that allowed the simultaneous assessment of their antagonist potency against phenylephrine-mediated increases in prostatic pressure and mean blood pressure. The intravenous administration of each of these compounds dose-dependently inhibited phenylephrine-induced increases in prostatic pressure and mean blood pressure. To further assess the ability of the three compounds to inhibit phenylephrine-induced responses, the doses required to produce a 50% inhibition of the phenylephrine-induced increases in prostatic and mean blood pressure and the selectivity index obtained from the ratio of those two doses were determined for each test compound. Forty minutes after the intravenous administration of naftopidil, the selectivity index was 3.76, and those of tamsulosin and prazosin were 1.23 and 0.61, respectively. These findings demonstrated that naftopidil selectively inhibited the phenylephrine-induced increase in prostatic pressure compared with mean blood pressure in the anesthetized dog model. The selectivity of naftopidil for prostatic pressure was the most potent among the test compounds. In addition, using cloned human alpha1-adrenoceptor subtypes, naftopidil was selective for the alpha1d-adrenoceptor with approximately 3- and 17-fold higher affinity than for the alpha1a- and alpha1b-adrenoceptor subtypes, respectively. The selectivity of naftopidil for prostatic pressure may be attributable to its high binding affinity for alpha1a- and alpha1d-adrenoceptor subtypes.

Adrenergic alpha-1 Receptor Antagonists↗

[Serum leptin level among non-obese students: relationship to body fat, blood pressure, serum lipids, physical activity, and eating habits].

Leptin, a product of the ob gene, is thought to play a key role in the regulation of adiposity. However, it is unclear in humans as to whether or not leptin influences the blood pressure, serum lipids, physical activity level, or eating behavior in relation to obesity. Recent reports have indicated both gender-based differences in the leptin levels and a correlation of the percentage of body fat with leptin levels has been observed among obese subjects. As far as we know, these relationships among non-obese young adults have yet to be studied. Therefore, the serum leptin concentrations among 107 non-obese students (72 males and 35 females) were measured by a radioimmunoassay(RIA). Fasting leptin levels ranging between 1.2 and 23.4 ng/mL were observed in all subjects, and the levels among females were 2.6 times higher than in males (7.64 vs 2.95 ng/mL; p < 0.001). A close correlation was observed regarding the log-transformed leptin levels with the percentage of body fat as determined by the bioelectrical impedance analysis method(r = 0.734, p < 0.001 in males and r = 0.579, p < 0.001 in females). In conclusion, these data thus suggest the serum leptin levels among non-obese students show significant gender-based differences while, in addition, the leptin levels also correlate positively with the percentage of body fat. However, it remains unclear as to whether or not the leptin levels are independently related to the blood pressure, serum lipids, physical activity level (sports activity and leisure-time activity), and eating behavior (eating breakfast, mid-day snacking and nighttime snacking).

Adipose Tissue↗

Effects of docosahexaenoic and eicosapentaenoic acid on lipid metabolism, eicosanoid production, platelet aggregation and atherosclerosis in hypercholesterolemic rats.

Exogenously hypercholesterolemic (ExHC) rats were fed on an atherogenic diet supplemented with 1% each of either ethyl ester docosahexaenoic acid [EE-DHA, 22:6(n-3)], ethyl ester eicosapentaenoic acid [EE-EPA, 20:5(n-3)] or safflower oil (SO) for 6 months. The rats fed on the diets containing EE-EPA or EE-DHA, compared with those fed on SO, had lower serum cholesterol and triacylglycerol levels, less aggregation of platelets and slower progress of intimal thickening in the ascending aorta. Relative to the SO-fed rats, both of the (n-3) fatty acid-fed rats had a significantly reduced proportion of arachidonic acid in the platelet and aortic phospholipids, and lower production of thromboxane A2 by platelets and of prostacyclin by the aorta. These results suggest that EPA and DHA are similarly involved in preventing atherosclerosis development by reducing hypercholesterolemia and modifying the platelet functions.

Animals↗

Concentration of serum lipids and aortic lesion size in female and male apo E-deficient mice fed docosahexaenoic acid.

Apolipoprotein (apo) E-deficient mice were fed an atherogenic diet with either 1% ethyl ester docosahexaenoic acid (DHA) or safflower oil (SO) as a source of linoleic acid for 8 week. Both genders fed DHA had higher proportions of eicosapentaenoic acid and DHA, and lower proportions of linoleic and arachidonic acids in the liver and serum phospholipids than those fed SO. Males fed DHA had greater liver weight and tended to have higher concentrations of serum lipids and liver cholesterol than those fed SO, and there were opposite trends in females. Dietary fats and gender led to no significant effect on lesion sizes in aortic arch and thoracic plus abdominal aorta. These results indicate that the interactive action of sex-related factor(s) with dietary polyunsaturated fatty acids is involved in metabolic changes of serum lipids in apoE-deficient mice, and addition of DHA, compared with addition of SO, is not effective to abolish the atherosclerosis in this animal model.

Animals↗

Diurnal variations of blood pressure in dogs.

Using the telemetry system, we measured the blood pressure (BP) invasively in seven adult mongrels while unanesthetized and unbound. Post-operative BP after implanting the telemetry BP transmitter showed temporarily high values due to the invasive nature of the surgery. It was, however, observed that BP gradually decreased thereafter, and showed settled trends from the eighth day post-operatively. When we took the average of the systolic, mean and diastolic BP at hourly intervals for each of the dogs once their BP had settled, a twin peak diurnal variation (at 8:00 and 19:00) was observed. Moreover, significantly high values (p < 0.05) were identified in active state compared with when sleeping or at rest. The 24 hr BP measured by the telemetry system in seven normal dogs resulted in the following values: systolic 123.4 +/- 7.9 mmHg, mean 91.1 +/- 5.6 mmHg, and diastolic 74.5 +/- 4.9 mmHg.

Animals↗

Tryptophan-niacin metabolism in liver cirrhosis rat caused by carbon tetrachloride.

We investigated the change of tryptophan-niacin metabolism induced by carbon tetrachloride (CCl4) in rats with liver cirrhosis. The rats were injected with CCl4 (0.5 or 1 mL of 50% olive oil solution/kg body weight) twice a week for 1 or 2 mo and given phenobarbital water simultaneously. The urinary excretions of nicotinamide (Nam) and its metabolites were assayed. As the result, the urinary excretion of Nam, N1-methyl-4-pyridone-3-carboxamide (4-Py), Nam + N1-methylnicotinamide (MNA) + N1-methyl-2-pyridone-5-carboxamide (2-Py) + 4-Py was lower in the CCl4-treated groups than in the non-treated group (control) regardless of the experimental period (1 mo and 2 mo) or dosing amount of CCl4 (0.5 and 1 mL). Moreover, we investigated which pathway of tryptophan-niacin metabolism was affected in CCl4-treated rat. As the result, the possibility that the MNA-->4-Py reaction is inhibited by CCl4 treatment was suggested in this experiment.

Alanine Transaminase↗

Automated filtration-based high-throughput plasmid preparation system.

Current methods of plasmid preparation do not allow for large capacity automated processing. We have developed an automated high-throughput system that prepares plasmid DNA for large-scale sequencing. This system is based on our previously reported filtration method. In this method, cell harvesting, alkaline lysis, and plasmid purification occur in a single 96-well microtiter plate from which sequence-ready DNA samples are collected. The plates are designed to allow all reagents to be injected from above the wells and the spent reagents to be aspirated from below. This design has enabled us to build a linear process plasmid preparation system consisting of an automated filter plate stacker and a 21-stage automated plasmid preparator. The 96-well plates used are outfitted with glass-filters that trap Escherichia coli before the plates are stacked in the automated stacker. The plates move from the stacker to each of the 21 stages of the preparator. At specific stages, various reagents or chemicals are injected into the wells from above. Finally, the plates are collected in the second stacker. The optimal throughput of the preparator is 40,000 samples in 17.5 hr. Here, we describe a pilot experiment preparing 15,360 templates in 160 specially designed 96-well glass-filter plates. The prepared plasmids were subjected to restriction digestion, DNA sequencing, and transcriptional sequencing.

Base Sequence↗

[Modulations by stimulation of angiotensin II type 1 receptor of gastrointestinal functions].

Electrolytes and water are bidirectionally transported across the epithelial cells of mammalian small and large intestines. In animals with chronic renal failure, plasma levels of angiotensin II and specific 125I-angiotensin II binding sites increase and result in enhanced excretions of K+, Cl- and urate from distal colon, in an AT1-receptor sensitive manner. Angiotensin-converting enzyme activity and mRNA levels for renin-2 and AT1A-receptor are blunted or lowered transiently by gastric intake of low Na+ in rodent, suggesting that angiotensin II serves as a humoral mediator for "gastric Na+ monitor". Angiotensin II causes also very potent contractions of gastrointestinal smooth muscles as shown in vascular smooth muscles. This review is focused on understanding potential roles of angiotensin II in gastrointestinal functions.

Angiotensin II↗

[Pulmonary radiation injury manifested by signs of bronchiolitis obliterans with organizing pneumonia after postoperative breast cancer radiotherapy].

A 67-year-old woman underwent surgery for cancer of both breasts (right: mastectomy, left: conserving surgery), and received 60 Gy radiation to the left postoperative breast. Three months later, cough and fever developed. A chest radiograph demonstrated infiltrative shadows in the left lung field. Transbronchial lung biopsy specimens disclosed organizing exudates in the alveolar spaces and bronchioles. After treatment with prednisolone, the clinical symptoms and radiographic infiltrates disappeared. This was a case of pulmonary radiation injury pathologically manifested by signs of bronchiolitis obliterans with organizing pneumonia.

Aged↗

Muscarinic acetylcholine receptor-mediated increase of angiotensin type 2 receptor mRNA in PC12 cells.

The present study examined changes in the angiotensin type 2 (AT2) receptor mRNA level after carbachol exposure in PC12 cells. The AT2 receptor mRNA level increased 2- to 3-fold after 12-18 h of carbachol exposure (100 microM). The up-regulation of AT2 receptor mRNA was antagonized by atropine, which is a muscarinic receptor antagonist, thus suggesting that the increase in AT2 receptor mRNA is mediated via muscarinic acetylcholine receptors. This increase is blocked by NG-nitro-L-arginine-methylester, nitric oxide (NO) synthase inhibitor, and hemoglobin NO trap. Protein kinase C (PKC) inhibitors, H-7 and calphostin C, inhibited the carbachol-induced upregulation. In addition, a G kinase inhibitor, ODQ (1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one), inhibited this increase. These findings suggest that the carbachol-induced increases in AT2 receptor mRNA is regulated by the activation of NO-cGMP and/or the PKC pathway.

Animals↗