Biomedical subjects

T Kokubu

Publications and source records attributed to T Kokubu.

At least 19 recordsLinked to original sources

PubMed2001

Isolation of chlamydia psittaci from domestic cats with oculonasal discharge in Japan.

Eight strains of Chlamydia psittaci were isolated in Japan from the nasal and conjunctival swabs of six household cats using the L929 cell line of mouse fibroblast origin. The isolates were identified as C. psittaci on the basis of the formation of characteristic inclusion bodies in the cell culture detected by Giemsa stain and immunofluorescence. Comparison of nucleotide sequences of the ompA gene amplified from the three isolates with the published sequence of feline FEPN strain of C. psittaci showed almost 100% homology.

Animals↗

PubMed2001

Low-intensity pulsed ultrasound initiates bone healing in rat nonunion fracture model.

Low-intensity pulsed ultrasound exposure has been shown clinically to shorten the fracture repair process and to induce healing of nonunions in humans, but its mechanism of action remains unclear. In this study we investigated the effect and mechanism of low-intensity pulsed ultrasound on nonunion fracture healing in rat tibias. A consistently reproducible nonunion was produced in rat tibias by muscle interposition without osteotomy. This model was produced by creating a closed tibial fracture with only the distal end of the tibialis anterior muscle interposed into the fracture site. One limb was noninvasively exposed to low-intensity pulsed ultrasound (a 200-millisecond burst of sine waves of 1.5 MHz, repeating at 1.0 kHz) for 20 minutes daily. The incident intensity was approximately 30 mW/cm2. Rats were killed at intervals between 2 and 6 weeks. The events were assessed by radiographs, microfocus X-ray computed tomograms, and histologic examination. After 6 weeks of exposure, 7 of 14 nonunion fractures showed healing on radiologic assessment. The results of three-dimensional microfocus X-ray computed tomographic reconstruction and histologic examination also supported this finding. On the other hand, all control tibias remained in a state of nonunion during the same period. These results indicate that low-intensity pulsed ultrasound promotes healing in the rat nonunion fracture model.

Animals↗

PubMed1999

Low intensity pulsed ultrasound exposure increases prostaglandin E2 production via the induction of cyclooxygenase-2 mRNA in mouse osteoblasts.

Both prostaglandin E2 (PGE2) and low intensity pulsed ultrasound (U/S) exposure have been reported to accelerate fracture repair. We hypothesized that these two pathways are interactive. To verify this hypothesis, we examined the regulation of PGE2 production by U/S exposure (sine wave of 1.5MHz repeating at 1kHz, 30mW/cm2, 20 minutes) in mouse osteoblastic cell line, MC3T3-E1. The production of PGE2 in osteoblasts was augmented by U/S, which was threefold at 60 min. in comparison with unexposed samples. Then we evaluated the expression of cyclooxygenase-2 (COX-2) mRNA, which is a critical enzyme for PGE2 production. U/S rapidly up-regulated the expression of COX-2 mRNA in a time dependent manner. In addition, PGE2 production by U/S was drastically suppressed by a selective inhibitor of COX-2. These results provide strong evidence that PGE2 production in osteoblasts is dependent upon the induction of COX-2 mRNA expression by U/S and offer a mechanistic insight into how U/S accelerates fracture repair.

Animals↗

PubMed1998

EGG phosphatidylcholine combined with vitamin B12 improved memory impairment following lesioning of nucleus basalis in rats.

We investigated the effects of egg phosphatidylcholine (PC) combined with vitamin B12 on memory in the Morris water maze task, and on choline and acetylcholine (ACh) concentrations in the brain of rats. Animals with nucleus basalis Magnocellularis (NBM) lesion received intragastric administration of egg PC or vitamin B12, or both for 18 days. Memory acquisition and retention were remarkably impaired in NBM lesioned rats compared with in sham-operated control. NBM lesioned group had lower choline and ACh concentrations than control group in the frontal cortex. High dose of egg PC alone significantly increased choline concentration, but did not change ACh concentration in the frontal cortex. High dose of vitamin B12 alone did not change choline and ACh concentrations in the brain. Either egg PC or vitamin B12 did not improve memory acquisition and retention. However, low dose of egg PC combined with vitamin B12 significantly increased ACh concentration and improved memory acquisition and retention in the NBM lesioned rats. We concluded that egg PC combined with vitamin B12 improved the memory impairment of NBM lesioned rats through the action on the cholinergic neurons.

Acetylcholine↗

PubMed1998

Efficacy of a mixed glycoprotein vaccine against pseudorabies in pregnant sows.

A pseudorabies virus (PRV) glycoprotein-mixed vaccine was prepared by heparin-affinity chromatography from PRV-infected PK-15 cell lysates. In our previous study, the trial vaccine was induced protection with suppression of virus shedding in one-month-old pigs and generation of cytotoxic T lymphocyte (CTL) response in mice. In this study, the effect of the trial vaccine on suppression of both virus shedding and reproductive failure in pregnant sows was examined. Three sows were vaccinated twice until one week before mating. Each of them was infected intranasally with 10(6) TCID50 of PRV on day 28, 54, or 85 after mating, respectively. Three other sows were also mated and challenged at the same time as the respective control. The vaccinated sows produced virus-neutralizing antibodies. Sows with high level of VN antibody lowered the level and period of virus shedding after challenge. The maximum level of shed-virus titers in vaccinated sows were 10(1.25) to 10(3) times lower than controls. Vaccinated sows shed virus for 1 or 5 days, while controls shed for 8, 9, or 12 days. No abortion or stillbirth was observed in vaccinated sows during pregnancy. On the other hand, control sow challenged at a late stage of pregnancy showed abortion and stillbirth. The results obtained here indicate that our trial vaccine is effective to prevent reproductive failure by pseudorabies virus.

Abortion, Veterinary↗

PubMed1998

Influence of cell surface glycoprotein gC produced by pseudorabies virus on cytopathic effect.

The wild-type pseudorabies virus (WT-PRV) produced a round-type cytopathic effect (CPE) in PK-15 cell line of porcine kidney origin, while PRVgCs lacking in gC-transmembrane-anchor region and PRVgC-defecting in gC gene produced a syncytium-type CPE. The mouse embryo cell line (BALB/3T3 clone A31) were transfected with recombinant plasmid of pcDNA3 which incorporated with gC gene. The transfected A31/gC cells were stably expressing gC. Only a round-type CPE was observed in these cells infected with WT-PRV, while a syncytium-type CPE was observed in the cells infected with each of the PRVgCs and PRVgC-. Any viruses described above induced a syncytium-type CPE in A31/pcDNA cells transfected with a plasmid without gC gene. By WT-PRV infection, PK-15 cells generated about 2- or 8-fold more gC than the A31/gC and A31/pcDNA cells when gC was measured by hemagglutination test. Flowcytometric analysis revealed that amount of gC on the cell surface of A31/gC and PK-15 cells increased after infection with WT-PRV. Round-type CPE was observed with the increase of gC. These results suggest that the type of CPE formation induced by PRV is dominated by the amount of gC on the infected cell surface.

3T3 Cells↗

PubMed1997

Special folding pathway to tetramer only through the micelle state of the corticotropin-releasing factor.

The ovine corticotropin-releasing factor (CRF), a peptide hormone of 41 residues stimulating the secretion of adrenocorticotropic hormone, was thermodynamically investigated. By means of size exclusion chromatography and/or ultrafiltration, the CRF solution could be separated into random coil monomers and highly alpha-helical tetramers, which seem to have amphipathic helix bundle structure. Circular dichroism measurements along with diluting or concentrating the CRF solution revealed that there exists the micelle state above the concentration of 0.1 mM, which would be the critical micelle concentration (cmc). The micelle state was also proved by binding ability for 8-anilino-1-naphthalenesulfonate and endothermic change by dilution across the cmc. The tetramer showed the cooperative thermal transition at about 55 degrees C in the buffer solution (pH 7.5), so that it would have native-protein-like folding. On the other hand, the micelle undergoes gradual change to dissociated state by heating, regardless of the similar alpha-helicity to the tetramer. Above the cmc the equilibrium between the tetramer and the micelle takes place as well as that between the monomer and the micelle. Whereas, the direct conversion between the tetramer and the monomer scarcely occurred below the cmc. The titration experiment with 2,2,2-trifluoroethanol (TFE) revealed that the cmc decreases with increasing the concentration of TFE. This tendency is the same as that of general surfactants. Most of experimental results can be well explained by this three-phase model involving the monomer, the tetramer, and the micelle. The lack of the equilibrium between the monomer and the tetramer indicates that the folding pathway of the tetramer is the transformation only through the micelle state and not from the monomer. This pathway resembles the collapse model among the phenomenological models for thermodynamic protein folding. By the mathematical consideration for the dissociation of micelle, we have demonstrated that the expected content of undegradable k-mer is 2/(k + 1), which agreed well with the observed tetramer content of CRF (40%).

Anilino Naphthalenesulfonates↗

PubMed1997

Bilateral adrenal lymphoma with neoplastic angioendotheliosis.

We report a case of bilateral lymphoma of the adrenal glands. A 61-year-old man was admitted to our hospital in clinical shock with anuria. Endocrine examination indicated a diagnosis of Addison's disease. Large tumors were found bilaterally in the suprarenal area, and a needle biopsy was performed, and the diagnosis was malignant lymphoma. Treatment with chemotherapy achieved a moderate response. He was discharged, but died 1 month later. On autopsy, microscopic examination showed extensive intravascular infiltration of lymphoma cells. We considered this to be a case of bilateral adrenal lymphoma with neoplastic angioendotheliosis.

Acute Kidney Injury↗

PubMed1996

Effects of dietary phosphatidylcholine on memory in memory deficient mice with low brain acetylcholine concentration.

Data concerning the effect of phosphatidylcholine (PCh) administration on the improvement of memory in senile dementia of Alzheimer type are inconsistent, probably due to the different conditions under which studies were conducted. Animal studies provide a good model, but data on this is limited. We studied the effect of PCh on memory in memory deficient mice (Dull mice) with low brain acetylcholine (ACh) concentration and normal mice. Mice were fed 24% casein diet (control) or this diet supplemented with 2 or 8% egg yolk PCh from gestation (Experiment 1) and after weaning (Experiment 2). Memory acquisition and retention were studied by step-down type passive avoidance performance at 8 and 10 weeks old, respectively. Control group of Dull mice had poorer memories than that of the normal mice in Experiments 1 and 2. On the 2% PCh diet, Dull mice improved memory acquisition and retention in Experiment 1 and retention in Experiment 2. On the 8% PCh diet in Dull mice there was improvement of memory and retention in Experiment 1, but no effect was observed in Experiment 2 (P > 0.05). In the normal mice, the 2% PCh diet did not affect memory acquisition and retention, however on the 8% PCh diet, there was no or adverse effect. These results suggest that dietary supplementation of egg yolk PCh improves memory of Dull mice, particularly when given from gestation and that the 2% PCh diet elicits better response than the 8% PCh diet.

Acetylcholine↗

PubMed1996

Genetic controls of susceptibility and resistance to 4-nitroquinoline 1-oxide-induced tongue carcinomas in rats.

We analyzed the incidence of infiltrative mass-type tongue carcinomas (IMTC) induced in 550 rats by continuous oral administration of 0.001% 4-nitroquinoline 1-oxide solution for 180 days. The study included various crosses of susceptible Dark-Agouti rats (DA) and resistant Wistar/Furth rats (WF). DA showed a 93.6% incidence of IMTC measuring more than 5 mm in their largest diameter, while WF showed only a 4% incidence. Reciprocal F1 and F2 hybrids mated by DA and WF showed 47.5% and 45.8% incidences, respectively. Meanwhile, reciprocal backcrossed hybrids to DA and WF showed 73.7%, and 24.6% incidences, respectively. Segregation of the incidences suggests that there are two autosomal dominant genes, one linked to the susceptibility of DA and the other to the resistance of WF.

4-Nitroquinoline-1-oxide↗

PubMed1996

Effect of egg white on serum cholesterol concentration in young women.

In a previous study we observed favorable effects of egg white on serum lipids in rats and mice. The present study was designed to elucidate these effects in 24 female university students with moderate hypercholesterolemia. About 30% of total protein was supplied with egg white, tofu or cheese. The experiment was conducted for a complete menstruation cycle of each subject. Lipid intake was about 30% of total energy intake. The energy intake of each subject was constant throughout the experiment. Body weight was measured every morning. Daily activity was measured by a pedometer. Blood was withdrawn after an overnight fast on the first, 15th and last days and serum lipids were measured. Body weight was measured every morning. Daily activity was measured by a pedometer. Blood was withdrawn after an overnight fast on the first, 15th and last days and serum lipids were measured. Body weight and daily activity were maintained in all the groups throughout the experiment. The egg white group showed a similar decrease in the total cholesterol (Total-C) concentration but a greater increase of high-density lipoprotein cholesterol (HDL-C) concentration as compared to the tofu group and a greater decrease in Total-C and low-density lipoprotein cholesterol (LDL-C) concentrations and a greater increase in the HDL-C concentration as compared to the cheese group (p < 0.05). The results indicate the favorable effects of egg white in the control of hypercholesterolemia.

Adult↗

PubMed1995

Administration of phosphatidylcholine increases brain acetylcholine concentration and improves memory in mice with dementia.

Studies on the effect of phosphatidylcholine administration on memory are limited. We administered egg phosphatidylcholine to mice with dementia and to normal mice and compared the differences in memory and serum choline concentration, and choline and acetylcholine concentrations and choline acetyltransferase activities of three forebrain regions (cortex, hippocampus and the remaining forebrain). Mice with dementia were produced by mating sibling mice who had impaired memory for > 20 generations. These mice had poor memory and low brain acetylcholine concentration. We administered 100 mg of egg phosphatidylcholine (phosphatidylcholine group) or water (control group) by gavage to each mouse daily for about 45 d. Control mice with dementia had poorer memory in passive avoidance performance and lower brain choline (cortex and hippocampus) and acetylcholine (hippocampus and forebrain excluding cortex and hippocampus) concentrations and lower cortex choline acetyltransferase activity than the control normal mice (P < 0.05). The administration of phosphatidylcholine to mice with dementia improved memory and generally increased brain choline and acetylcholine concentrations to or above the levels of the control normal mice. In normal mice, phosphatidylcholine treatment did not affect memory or acetylcholine concentrations in spite of the great increase in choline concentrations in the three brain regions. Serum choline concentration in mice treated with phosphatidylcholine increased to a similar level in both strains of mice, indicating that the absorption of phosphatidylcholine was not impaired in mice with dementia. The results suggest that administration of egg phosphatidylcholine to mice with dementia increases brain acetylcholine concentration and improves memory.

Acetylcholine↗

PubMed1995

Production of hetero-dimeric dihydrofolate reductase-thymidylate synthase bifunctional enzyme.

As a result of the expression of a single open reading frame composed of the coding sequence for a cysteine-free mutant (Cys85-->Ala, Cys152-->Ser) of Escherichia coli dihydrofolate reductase (DHFR; 18K monomeric protein), that for the E. coli thymidylate synthase (TS; dimeric protein with a 30K promoter), and a spacer sequence (coding 7 amino acids) with a Shine-Dargarno sequence, an active hetero-dimeric bifunctional enzyme with 50K DHFR-TS and 30K TS polypeptides was stably produced in the transformed E. coli cell in addition to an overproduction of the TS dimeric enzyme. The highly purified hetero-dimeric enzyme has similar Vmax and Km values in both DHFR and TS activities to those of the natural counterparts, monomeric DHFR and dimeric TS. Although the hetero-dimeric enzyme did not show an apparent channeling transfer of dihydrofolate (the intermediate substrate) between the spatially discrete DHFR and TS active sites, the coupling efficiency of the TS and DHFR reactions in the artificial enzyme was better than that in the separated enzymes, as shown by a decrease in the intermediate concentration at the steady state in the coupled reaction.

Amino Acid Sequence↗

PubMed1995

Blood pressure response to the Valsalva maneuver in pheochromocytoma and pseudopheochromocytoma.

To elucidate whether a difference in blood pressure reactivity exists between patients with pheochromocytoma (n = 8) and pseudopheochromocytoma (n = 22), we evaluated blood pressure changes during a Valsalva maneuver and baroreceptor reflex sensitivity. We also examined the effects of propranolol and prazosin on blood pressure reactivity during a Valsalva maneuver in patients with pseudopheochromocytoma. Pseudopheochromocytoma was defined as a paroxysmal rise in blood pressure accompanying pheochromocytoma-like symptoms and normal catecholamine values. The difference in systolic blood pressure between phase IV of the Valsalva maneuver and baseline (delta SBP) was markedly smaller in the pheochromocytoma patients (8.4 +/- 18.4 mm Hg) than in the essential hypertension patients (n = 30, 30.9 +/- 19.4 mm Hg) and normotensive control subjects (n = 10, 31.3 +/- 11.4 mm Hg), whereas delta SBP in the pseudopheochromocytoma patients (77.8 +/- 11.2 mm Hg) was markedly greater than in the other three groups. delta SBP was markedly suppressed by the administration of both propranolol and prazosin. Baroreceptor reflex sensitivity index was lower in the pheochromocytoma group than in the other three groups. In conclusion, blood pressure reactivity responses to a Valsalva maneuver are disparate between pheochromocytoma and pseudopheochromocytoma. The high blood pressure reactivity to a Valsalva maneuver in pseudopheochromocytoma is due to hyperactivity in both beta- and alpha 1-adrenergic receptor functions, and the low blood pressure reactivity to a Valsalva maneuver in pheochromocytoma seems to be mainly due to the desensitization of both adrenergic systems associated with chronic catecholamine excess. In addition, the impaired baroreceptor function in pheochromocytoma is partially responsible for it.

Adrenal Gland Neoplasms↗

PubMed1995

Vasospastic angina in two sisters.

Vasospastic angina was observed in two sisters. The 52-year-old younger sister presented with rest angina at midnight and in the early morning. The coronary arteriogram showed no significant organic stenosis. Vasospasm to the left anterior descending and right coronary arteries was induced by the intracoronary administration of acetylcholine. The 57-year-old elder sister complained of rest and effort angina. Her coronary arteriogram was also normal. Vasospasm to the left circumflex and right coronary arteries was provoked by acetylcholine. In both cases, human leukocyte antigen DQw3 was negative. In the present cases, genetic factors may partly contribute to the mechanism of vasospastic angina.

Acetylcholine↗

PubMed1995

Detection of perioperative coronary vasospasm on iodine-123-MIBG, thallium-201 and iodine-123-BMIPP myocardial SPECT images.

A 64-yr-old woman, who had no history of anginal attack, underwent cholecystectomy under general anesthesia. A few hours after successful surgery, the EKG showed T-wave inversions with QT-interval prolongations and torsades de pointes. The [123I]MIBG scintigraphic findings revealed marked denervation in the anteroseptal and inferoposterior myocardium, whereas 201TI myocardial SPECT showed only slightly reduced tracer uptake in those areas. The [123I]BMIPP scans showed abnormal fatty acid metabolism in the anteroseptal myocardium. Coronary angiogram detected no fixed stenosis, but coronary vasoconstriction in the left anterior descending artery was induced by intracoronary injection of acetylcholine. In our patient, the findings of cardiac imaging with [123I]MIBG, 201TI and [123I]BMIPP led to coronary angiography and the final diagnosis of probable perioperative coronary vasospasm.

3-Iodobenzylguanidine↗

PubMed1995

Effect of age on left ventricular geometric patterns in hypertensive patients.

OBJECTIVES: To investigate the effect of age on left ventricular structure and geometry in hypertensive patients, we studied the relationship between age and echocardiographic variables in patients with uncomplicated essential hypertension. PATIENTS AND METHODS: We divided 168 patients with hypertension into three groups according to age: young (<40 years), middle-aged (40-59 years) and an elderly group (> or = 60 years). They were further categorized according to relative wall thickness and the left ventricular mass index. We then evaluated the prevalence of left ventricular geometric patterns in these patients according to age. RESULTS: The left ventricular end-diastolic dimension decreased with age, both in normotensive control subjects and in hypertensive patients. The magnitude of this decrease was similar for both. The relative wall thickness and left ventricular mass index were greater in the hypertensive patients than in the normotensive control subjects, and these increased with age both in the controls and the hypertensives. The differences between normotensives and hypertensives in these variables remained unchanged with age. The prevalence of a normal left ventricle (normal relative wall thickness and left ventricular mass index) in the hypertensive patients decreased with age. Conversely, the prevalence of concentric remodeling (increased relative wall thickness with normal left ventricular mass index) and concentric hypertrophy (increased relative wall thickness and left ventricular mass index) increased with age. CONCLUSIONS: These results demonstrate that age significantly affects left ventricular structure both in normotensive control subjects and in hypertensive patients. Thus, the differences in left ventricular geometric patterns with age may have important implications in assessing left ventricular structure and geometric patterns in hypertensive patients.

Adult↗

PubMed1994

Gallium-67 citrate scintigraphy in idiopathic pericarditis--report of a case.

We report a case of idiopathic pericarditis in a 71-year-old woman without a friction rub or electrocardiographic changes suggestive of pericardial inflammation. She noticed dyspnea and palpitation on exertion about 40 days before admission. After admission, pericardial effusion and inflammatory reactions, such as elevated C-reactive protein, were found. Moreover, gallium-67 citrate scintigraphy revealed abnormal isotope accumulation over the cardiac silhouette. Therefore, she was diagnosed as having "active" pericarditis. The finding with the gallium scan became negative with steroid therapy. In this case, a gallium scan was very useful in assessing and monitoring the course of pericardial inflammation.

Aged↗