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Biomedical subjects

M Tokunaga

Publications and source records attributed to M Tokunaga.

At least 37 records · Page 2Linked to original sources
PubMed2001

Epstein-Barr virus involvement is mainly restricted to lymphoepithelial type of gastric carcinoma among various epithelial neoplasms.

To demonstrate the association of Epstein-Barr virus (EBV) with primary epithelial neoplasms in the south part of Kyushu, Japan, 761 carcinomas consisting of 75 lung, 61 breast, 107 esophagus, 102 colon, 58 pancreas, 45 thyroid, and 313 gastric cancers were examined by EBER-1 in situ hybridization. EBER-1 was detected in 23 cases (7.3%) out of 313 gastric carcinomas, while none of the other carcinomas was positive for EBER-1. Twenty-eight (9.4%) out of 313 gastric carcinomas were differentiated poorly to moderately carcinomas with prominent lymphoid cell infiltration, similar to so-called lymphoepithelioma-like carcinoma, and 19 cases (67.9%) were positive for EBER-1. Although two (2.6%) and 11(10.3%) out of 75 lung and 107 esophagus carcinomas were so-called lymphoepithelioma-like carcinomas, respectively, but EBER-1 was not detected in other epithelial neoplasms that originated from the lung, esophagus, breast, colon, pancreas, and thyroid in the south of Kyushu, Japan. As a result, EBV was associated with only some gastric carcinomas but not with other epithelial neoplasms originating from the lung, esophagus, breast, colon, pancreas, and thyroid in southern Japan.

Adenocarcinoma↗
PubMed2001

Efficient expression, purification and characterization of mouse salivary alpha-amylase secreted from methylotrophic yeast, Pichia pastoris.

We constructed a secretion vector of mouse salivary alpha-amylase, pPAM, using the AOX1 promoter-terminator and the secretion signal of 128 kDa pGKL killer protein, for an alternative yeast, Pichia pastoris. Taking advantage of multicopy insertion of the expression cassette and optimized growth conditions, we succeeded in highly efficient extracellular production (approximately 240 microg/ml) of mouse alpha-amylase in the 10 ml scale by conventional flask culture: this efficiency was about 90-fold higher than that of Saccharomyces cerevisiae. Growth temperature of cells was critical for efficient production of alpha-amylase. P. pastoris transformants secreted both core-glycosylated and non-glycosylated alpha-amylase molecules with a glycosylated:non-glycosylated ratio of about 20:80. Both glycosylated and non-glycosylated alpha-amylases were purified separately to apparent homogeneity. The signal sequence was correctly processed in both species, and the molecular masses of glycosylated and non-glycosylated alpha-amylase were determined to be 58 600 and 56 300, respectively, by mass spectrometry. We further studied the outer chain glycosylation of engineered mouse alpha-amylase secreted by P. pastoris.

Amino Acid Sequence↗
PubMed2001

Expression and purification of cytokine receptor homology domain of human granulocyte-colony-stimulating factor receptor fusion protein in Escherichia coli.

Direct expression of the cytokine receptor homology (CRH) domain of granulocyte-colony-stimulating factor (G-CSF) receptor is lethal to Escherichia coli. For the efficient and stable production of an active CRH domain in E. coli, we fused the CRH domain with different proteins, such as maltose-binding protein (MalE), glutathione S-transferase, and thioredoxin (Trx). Among these, Trx appeared to be the best in terms of the protein expression level, purification efficiency by affinity chromatography, and binding activity to its ligand, G-CSF. The yield of active Trx-CRH fusion protein increased about 200-fold compared to that of previously reported MalE-CRH fusion.

ATP-Binding Cassette Transporters↗
PubMed2001

Expression, purification, and characterization of the active immunoglobulin-like domain of human granulocyte-colony-stimulating factor receptor in Escherichia coli.

We succeeded in the expression, purification, and refolding of the immunoglobulin-like (Ig) domain of human granulocyte-colony-stimulating factor (G-CSF) receptor with amino-terminal His-tag in Escherichia coli. The refolded Ig domain bound to a G-CSF affinity column and could be eluted with free G-CSF as a receptor-ligand complex, demonstrating that the Ig domain has the information necessary for binding its ligand, G-CSF. The eluted His-Ig/G-CSF complex could be separated from excess G-CSF by Ni-NTA column chromatography. The yield of this active recombinant His-Ig protein is about 0.72 mg per liter of culture. Its small size and the ease of production make this receptor fragment a useful reagent for the structural analysis of its complex with G-CSF.

Amino Acid Substitution↗
PubMed2001

An assessment of a method of inflating cuffs with a nitrous oxide gas mixture to prevent an increase in intracuff pressure in five different tracheal tube designs apparatus.

We assessed the efficacy of inflating cuffs with a nitrous oxide gas mixture to minimise changes in intracuff pressure during anaesthesia. Patients were randomly assigned to one of five groups of 15 subjects each, and the trachea was intubated with the Profile Soft-Seal Cuff, Hi-Contour, Reinforced, Sheridan or Trachelon tracheal tubes. Cuffs were inflated with 40% nitrous oxide and cuff pressure was measured during anaesthesia with 67% nitrous oxide. Concentration of nitrous oxide in the cuff was measured at the end of anaesthesia. Cuff pressure increased slightly but significantly in the Reinforced and Trachelon groups. Nitrous oxide concentration in the Reinforced, Sheridan, or Trachelon groups was slightly but significantly higher than that in the Profile or Hi-Contour groups. Cuff pressure never exceeded 22 mmHg and there were no air leaks. Therefore, inflating cuffs with 40% N2O preserves stable cuff pressure in all five tracheal tubes, despite differences in cuff and pilot balloon design.

Adolescent↗
PubMed2001

Human leukocyte antigens related to Epstein-Barr virus-associated gastric carcinoma in Japanese patients.

To assess the association between specific types of human leukocyte antigen and the risk of Epstein-Barr virus (EBV)-associated gastric carcinoma, serological typing for major histocompatibility complex class I and class II antigens was performed for 110 EBV-positive and 155 EBV-negative gastric carcinoma cases. In class I analysis, the frequency of B59 in the EBV-positive cases was higher than for the EBV-negative cases (odds ratio (OR) 3.06; 95% confidence interval (CI) 1.02-9.23). For class II antigens, DQ3 and DR9 frequencies in the EBV-positive cases were higher (OR 1.94; 95% CI 1.16-3.24 and OR 1.93; 95% CI 1.11-3.37, respectively), whereas DR11 frequency was lower than found in the EBV-negative cases (OR 0.10; 95% CI 0.01-0.79). After adjusting for multiple comparisons, only DR11 frequency remained significantly lower in the EBV-positive cases (P = 0.04), and the association of DQ3 was marginally significant (P = 0.05). These results suggest that the presence of DR11-restricted cytotoxic T cells (CTLs) related to EBV-associated gastric carcinoma, or a deficiency of DR11 and a high frequency of DQ3 may be genetic markers for a population at greater risk of EBV-associated gastric carcinoma. However, further extensive studies to more cases and DNA typing are needed because our findings in this study are exploratory.

Epstein-Barr Virus Infections↗
PubMed2001

Epstein-Barr virus-associated gastric carcinoma in Japanese Brazilians and non-Japanese Brazilians in São Paulo.

The proportion of Epstein-Barr virus-associated gastric carcinoma (EBV-GC) was examined in 149 Japanese-Brazilian and 151 non-Japanese-Brazilian gastric-carcinoma cases using in situ hybridization (ISH) assay to detect EBV-encoded small RNA (EBER), and the results were compared with our referent Japanese data. We found that 4.7% of cases in Japanese Brazilians were EBER-positive. This frequency was slightly lower than that of the referent Japanese, among whom 6.2% of 2038 gastric-carcinoma cases were EBER-positive. On the other hand, the non-Japanese-Brazilian series showed a significantly higher proportion of EBV-GC (11.2%) than the referent group did (P = 0.01). Although EBV-GC was predominant in males among non-Japanese Brazilians (M / F = 3.6, P = 0.047), as was the case in Japanese (M / F = 2.7), Japanese Brazilians did not show such a male predominance. The sex-ratio difference between the Japanese Brazilians and Japanese was statistically significant (P = 0.005). In conclusion, the present study in Japanese Brazilians and Japanese yielded no evidence suggesting any change in the frequency of EBV-GC caused by migration, except the absence of male predominance, which was observed both in Japanese and non-Japanese Brazilians.

Adenocarcinoma↗
PubMed2001

Liver cancer in atomic-bomb survivors: histological characteristics and relationships to radiation and hepatitis B and C viruses.

Histological features of primary liver cancer among atomic-bomb survivors and their relationship to hepatitis B (HBV) and C viral (HCV) infections are of special interest because of the increased risk of liver cancer in persons exposed to ionizing radiation and the high and increasing liver cancer rates in Japan and elsewhere. We conducted a pathology review of liver cancers occurring from 1958 to 1987 among subjects in the 120,321 member cohort of 1945 Hiroshima and Nagasaki residents. A panel of pathologists classified tumor histological types and defined accompanying cirrhotic changes of the liver. Archival tissue samples were assessed for HBV using pathology stains and PCR. Reverse transcriptase (RT) PCR was used to determine HCV status. We used unconditional logistic regression to compare 302 hepatocellular carcinoma (HCC) cases to 53 cholangiocarcinoma (CC) cases, adjusting for age, year of diagnosis, sex and viral status. Cirrhotic changes occurred significantly more often among HCC than CC cases (76% in HCC and 6% in CC). Compared to CC cases, HCC cases were 10.9 times more likely to be HBV-positive (95% confidence interval: 2.1-83.2) and 4.3 times more likely to be HCV-positive (95% confidence interval: 1.1-20.5). No significant differences were found between HCC and CC cases in radiation exposures. The predominance of HCC in the atomic-bomb survivors follows the background liver cancer pattern in Japan. Our findings suggest that HBV and HCV are involved in the pathogenesis of HCC with or without cirrhosis and are significantly less important in that of CC.

Aged↗
PubMed2001

Molecular cloning of groESL locus, and purification and characterization of chaperonins, GroEL and GroES, from Bacillus brevis.

The groESL locus of a protein-hypersecreting bacterium, Bacillus brevis, was cloned by PCR using primers designed based on the DNA sequence of a B. subtilis homolog. GroEL protein was purified to apparent homogeneity and its ATPase activity was characterized: it hydrolyzed ATP, CTP, and TTP in this order of reaction rate, and its specific activity for ATP was 0.1 micromole/min/mg protein. Purified GroEL forms a tetradecamer. GroEL was estimated to contain 22% alpha-helix, 24% beta-sheet, and 19% turn structures, by CD measurement. GroES protein was also highly purified to examine its chaperonin activity. GroEL protected from thermal inactivation of and showed refolding-promoting activity for malate dehydrogenase, strictly depending on the presence of ATP and GroES.

Adenosine Triphosphatases↗
PubMed2001

Characterization of nucleoside diphosphate kinase from moderately halophilic eubacteria.

Nucleoside diphosphate kinase was purified to apparent homogeneity from naturally isolated moderately halophilic eubacteria by ATP-agarose and phenyl-5PW column chromatographies. The molecular mass of this enzyme was 15 kDa by time-of-flight mass-spectrometry. This protein showed anomalous mobility on SDS-PAGE which is typical of a halophilic protein. It was stable and active over a wide range of salt concentrations, from 0 to 4.0 M NaCl.

Amino Acid Sequence↗
PubMed2001

Natural history of scoliosis in children with syringomyelia.

We performed a retrospective review of 27 scoliotic patients with syringomyelia using MRI. Their mean age at the first MRI examination was 10.9 years, and at the final review 15.8 years. The mean ratio of the diameter of the syrinx to the cord on the midsagittal MRI (S/C ratio) decreased from 0.49 to 0.24; 14 patients showed a decrease of 50% or more (reduction group). In this reduction group, the cerebellar tonsillar herniation decreased from a mean of 11.3 mm to 6.0 mm, and some improvement in dissociated sensory disturbance was seen in nine of 13 patients. The scoliosis improved by 5 degrees or more in six patients in the reduction group. Our results indicate that spontaneous shrinkage of syringomyelia in children is not unusual and is associated with improvement in the tonsillar herniation, the scoliosis and the neurological deficit.

Adolescent↗
PubMed2001

The effect of short-term (10- and 15-min) running at self-selected intensity on mood alteration.

The primary purpose of this study was to contrast the effects of two short-duration exercises on mood changes. A secondary goal was to examine the relationship between pre-exercise and post-exercise mood states. The subjects were 15 healthy male graduate students. They were involved in a within-subject design in which each individual completed two trials of running on a treadmill, one trial for 10 minutes and the other for 15 minutes. The Mood Checklist Short-form 1 (MCL-S1) used in the present study represents the participants' mood states before, during, and after exercise. This questionnaire has three sub-scales measuring (a) pleasantness, (b) relaxation, and (c) anxiety. Participants ran on a treadmill for the assigned time at a self-selected intensity after being told to run at a rate that felt good and was not painful. ANOVA results showed that both of the short bouts of exercise affected the subjects' mood, because the main effect of time spent exercising was observed in all sub-scales of the MCL-S1 and there were no significant differences in trial-by-time interaction. In addition to these results, there was a significant correlation between the two trial lengths in the amount of pleasantness and the amount of anxiety felt at post-exercise. There were moderate differences in the effect size (ES) for pre- and post-exercise pleasantness and anxiety levels. These results revealed similar patterns of change. It seems reasonable to conclude, based on this study, that exercise between 10 and 15 minutes results in similar psychological benefits for the person exercising.

Adult↗
PubMed2001

The relationships among exercise behavior, functional ADL, and psychological health in the elderly.

The purpose of this study was to examine the relationships among exercise behavior, the ability to live independently as assessed by functional activities of daily living (ADL), and psychological health as measured by the Philadelphia Geriatric Center morale scale, using data from 202 elderly residents aged 70 and living in their own home. The main results obtained from the analysis were as follows. 1. Exercise behavior had a significant effect on functional ADL. In particular, subjects who had been exercising regularly for 6 months or longer had higher functional ADL than subjects who did not exercise. 2. For psychological health, the functional ADL level was statistically significant, whereas exercise behavior was not. 3. Multiple regression analysis showed a positive and significant association between psychological health and functional ADL, but not between psychological health and exercise behavior, gender or family type. The results of this study demonstrated that elderly people's efforts to maintain and enhance their psychological health play important roles in their ability to live independently, and that regular exercise is necessary to an elderly person's ability to live independently.

Activities of Daily Living↗
PubMed2001

Bilateral massive pleural effusions caused by uremic pleuritis.

A 61-year-old man was started on hemodialysis in June 1998. Just after the commencement of dialysis, a chest X-ray film revealed bilateral pleural effusions. The effusions were hemorrhagic and exudative, and did not respond to dialysis. He was transferred to our university hospital on October 8,1998. Repeated thoracentesis demonstrated hemorrhagic and exudative characteristics without any diagnostic evidence. Pleural biopsies showed fibrosis and lymphocyte infiltration. The effusions were massive and did not respond to treatments including hemodialysis, repeatedly performed pleurodesis and the administration of antituberculous drugs. He died of respiratory failure on December 30, 1998. The autopsy confirmed bilateral fibrinous pleuritis without any underlying infections or malignancy. We diagnosed this case as uremic pleuritis from this clinical course and the autopsy findings. The clinical entity of uremic pleuritis was recognized as a complication of patients with hemodialysis in 1969. Uremic pleuritis generally responds to continued hemodialysis and the prognosis is usually good. However, some case reports demonstrated that surgical decortication is only indicated in cases with a severe clinical course. The clinical course of the present case was progressive and fatal. Uremic pleuritis is a serious complication of hemodialysis, which may lead to death.

Fatal Outcome↗
PubMed2001

Localized activation of m-calpain in migrating human umbilical vein endothelial cells stimulated by shear stress.

Using a parallel-plate flow-chamber and confocal laser scanning microscopy (CLSM), we studied the mode of cytoskeletal reorganization in migrating HUVECs stimulated by shear stress. Activation of m-calpain associated with a change in the spatial distribution of cytoplasmic ionized Ca2+ concentration ([Ca2+](i)) was studied. Shear stress (10 dyne/cm(2)) caused migration and decrease in the F-actin content of HUVECs. Migrating individual HUVECs showed the lamellipodium formed in the direction of cell migration, in which [Ca2+](i) elevated to 148 +/- 12 nM in a localized fashion. We found the appearance of activated m-calpain in the local area of the migrating HUVECs, which was associated with a decrease in the amounts of pp125FAK and ezrin. The localized rise in [Ca2+](i) might be closely related to morphological change to regulate the direction of cell migration induced by shear stress through localized activation of m-calpain.

Aniline Compounds↗
PubMed2000

Single-motor mechanics and models of the myosin motor.

Recent progress in single-molecule detection techniques is remarkable. These techniques have allowed the accurate determination of myosin-head-induced displacements and how mechanical cycles are coupled to ATP hydrolysis, by measuring individual mechanical events and chemical events of actomyosin directly at the single-molecule level. Here we review our recent work in which we have made detailed measurements of myosin step size and mechanochemical coupling, and propose a model of the myosin motor.

Actins↗
PubMed2000

Vertebral decancellation for severe scoliosis.

STUDY DESIGN: The results of staged surgery including vertebral decancellation were reviewed retrospectively for 21 patients with severe scoliosis. OBJECTIVES: To evaluate the benefits and limitations of vertebral decancellation as new anterior surgical procedure. SUMMARY OF BACKGROUND DATA: The curvatures of severe scoliosis are often very rigid, and surgical correction using the anterior or posterior approach may not achieve the desired correction. Some studies reported neurologic complications might appear due to the aggressive approach or excessive correction force. METHODS: Twenty-one patients (average age, 17.0 years) with severe scoliosis, in whom Cobb angle was over 80 degrees (average angle, 107 degrees), underwent staged anterior and posterior spinal reconstruction. Vertebral decancellation was performed as anterior procedure, and until posterior instrumentation, halo traction was carried out. The transition of curvatures in coronal and sagittal planes was assessed in this series. RESULTS: The average correction rate of lateral curvature at the final follow-up was 46%. The average loss of correction was 2.5 degrees. Kyphosis, measured between T5 and T12, changed from 41 degrees to 36 degrees. Lordosis, measured between L1 and S1, changed from 56 degrees to 45 degrees. Transient neurologic deficit was seen in one case after vertebral decancellation. CONCLUSIONS: Staged surgery including vertebral decancellation is an effective surgical method for patients with severe scoliosis, where an inflexible rigid curve or the risk of occurrence of neurologic complications due to temporary correction may exist.

Adolescent↗