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Biomedical subjects

N Tanaka

Publications and source records attributed to N Tanaka.

At least 145 records · Page 8Linked to original sources
PubMed2001

Mikimopine synthase (mis) gene on pRi1724.

By determination of the nucleotide sequence adjacent to the right border of T-DNA of the mikimopine-type Ri plasmid (pRi1724) in Agrobacterium rhizogenes, a new open reading frame (ORF) encoding 318 amino acids was found. A transcript of 1.35 kb derived from this ORF was observed in hairy roots of Ajuga reptans by northern blotting analysis. Including its own promoter and terminator, this ORF was isolated from the pRi1724 T-DNA and introduced into tobacco plants by the Agrobacterium-binary vector system. Since mikimopine, an opine and a stereoisomer of cucumopine, was accumulated in all organs of the transgenic tobacco plants, the new ORF was deduced to be the mikimopine synthase gene. For comparison, the nucleotide sequence of cucumopine synthase encoded on pRi2659 was also determined. No homology was found between mikimopine synthase and cucumopine synthase at the nucleotide, but partial homology was found at the amino acid level. Mikimopine synthase and cucumopine synthase produced by a protein expression system using E. coli catalyzed the synthesis of mikimopine and cucumopine from L-histidine and alpha-ketoglutaric acid, requiring NADH as a cofactor. These synthesized opines were identified by paper electrophoresis, TLC and HPLC analyses. The synthesized mikimopine or cucumopine could be degraded by A. rhizogenes strains harboring Ri plasmids encoding the respective catabolic enzyme.

Amino Acid Sequence↗
PubMed2001

Neonatal phencyclidine treatment selectively attenuates mesolimbic dopamine function in adult rats as revealed by methamphetamine-induced behavior and c-fos mRNA expression in the brain.

One of the major hypotheses regarding the pathogenesis of schizophrenia is the implication of neurodevelopmental abnormality. However, the mechanism of delayed onset of schizophrenic symptoms, in which increased dopaminergic activity in mesolimbic or mesocortical dopamine systems plays a pathological role, is not known. In this study, we investigated whether the chronic blockade of N-methyl-D-aspartate (NMDA) receptor by phencyclidine (PCP), an NMDA channel blocker, during development could disrupt the dopamine system during later life. Neonatal rats were injected with PCP subcutaneously daily from postnatal day (PD) 1 to PD 14 and their dopaminergic function was evaluated on PD 42 by rating the methamphetamine (MAP)-induced behavior. To illustrate the activated brain regions, the expression of c-fos mRNA in response to a MAP challenge was also studied utilizing in situ hybridization. Chronic neonatal PCP treatment attenuated MAP-induced oral stereotypy (licking and gnawing) and reduced MAP-induced expression of c-fos mRNA in the N. accumbens shell region and VTA but not in the N. accumbens core region, medial striatum, or substantia nigra. These results suggest that neonatal blockade of NMDA receptor, which induces a number of effects in the developing nervous system, may cause long-lasting functional changes of the mesolimbic dopamine system.

Animals↗
PubMed2001

Antiviral response by natural killer cells through TRAIL gene induction by IFN-alpha/beta.

Natural killer (NK) cells play an important role in early defense against viral infection. The cytotoxic activity of NK cells is increased by interferon-alpha/beta (IFN-alpha/beta), produced en masse in virally infected cells. However, the mechanism(s) by which IFN-alpha/beta contribute to the NK-cell-mediated antiviral response is not well understood. Here we provide evidence that the cytotoxicity of NK cells is enhanced by IFN-alpha/beta through induction of TNF-related apoptosis-inducing ligand (TRAIL). Isolation and analysis of the murine TRAIL promoter revealed the presence of an IFN-stimulated response element (ISRE), which binds to the transcription factor ISGF3 (interferon stimulated gene factor-3). This promoter is indeed activated by IFN-beta in ISGF3-dependent manner. We also show that virally infected cells, but not uninfected cells, are susceptible to TRAIL-mediated cytotoxicity in vitro, and that the TRAIL expressed in NK cells is indeed crucial in limiting virus replication in vivo. Thus, our study reveals a new molecular link between IFN-alpha/beta signaling and activation of NK cells in antiviral response of the host.

Animals↗
PubMed2001

Alterations in IRF1/IRF2 expression in acute myelogenous leukemia.

The interferon response genes 1 and 2 have been shown to be involved in the regulation of differentiation and proliferation of cells of the myeloid series, with the former functioning as an anti-oncogene and the latter as an oncogene. In the study described here, the levels of expression of these two genes and the ratio of their expression were compared in AML and normal marrow. The ratio of gene expression was significantly less in AML marrow cells as compared to normal marrow cells [med ratio = 1.33 vs. 2.97, P = 0.003]. While the expression ratio was unaffected by the presence or absence of either ras or fms mutations, p53 mutations were associated with higher IRF1:IRF2 expression ratios that wt p53 genes [med = 1.701 vs. 1.135, P = 0.014]. Given the functional characteristics and the competitive nature of these two genes, it is possible that leukemic transformation is associated with a fall in IRF1:IRF2 ratios. Finally, the administration of IL4 can result in the normalization of the IRF1:IRF2 ratio in the marrow cells of some patients with AML.

Adult↗
PubMed2001

Histopathologic impact of TT virus infection on the liver of type C chronic hepatitis and liver cirrhosis in Japan.

The present investigation compared the histological findings in the liver of chronic hepatitis C patients who were or were not co-infected with TT virus (TTV) to determine the histological and clinical characteristics of TTV infection. One hundred eighty patients with chronic hepatitis or liver cirrhosis type C were included in this study. Serum samples were tested for the presence of TTV DNA by a nested polymerase chain reaction. The liver biopsy specimen of each patient was examined, and scores were assigned to indicate the severity of each of the following features: inflammatory cell infiltration in the periportal, parenchymal, and portal areas; fibrous stage; lymphoid reaction in the portal area; portal sclerotic change; perivenular fibrosis; pericellular fibrosis; damage of bile duct; and irregular regeneration of hepatocytes. Sixty-four (34.4%) of the 180 patients were positive for TTV DNA. The histological features of the liver and the blood biochemical parameters of the TTV DNA-positive and TTV DNA-negative patients, did not differ significantly except for the score of irregular regeneration (IR) of hepatocytes. Among those in the F4 stage of fibrosis, the score of IR of the TTV DNA-positive patients was significantly higher than that of the TTV DNA-negative patients. In conclusion, chronic TTV infection does not modify the biochemical features of chronic hepatitis type C patients. TTV may be a risk factor, however, for the development of hepatocellular carcinoma in patients with type C liver disease in the F4 stage.

Adolescent↗
PubMed2001

Functional characterization of Gms1p/UDP-galactose transporter in Schizosaccharomyces pombe.

Galactosylation of glycoproteins in the fission yeast Schizosaccharomyces pombe requires the transport of UDP-galactose as substrate for the galactosyltransferase into the lumen of the Golgi apparatus, which is achieved by the UDP-galactose transporter. We isolated a mutant (gms1) that is deficient in galactosylation of cell surface glycoproteins in Sz.pombe, and found that the gms1(+) gene encodes a UDP-galactose transporter. In the prediction of secondary structure of the Gms1 protein, an eight-membrane-spanning structure was obtained. Fluorescent microscopy revealed the functional Gms1-GFP fusion protein to be stably localized at the Golgi membrane. Sequencing analysis of the coding region of Gms1p derived from galactosylation-defective mutants identified a single amino acid mutation (A102T or A258E) located within the putative transmembrane region, helix 2 or helix 7, respectively. The mutagenized Gms1(A102T or A258E)p exhibited loss of UDP-galactose transport activity but no change in the localization to the Golgi membrane. The C-terminal truncated Gms1p mutants demonstrated that the C-terminal hydrophilic region was dispensable for targeting and function as UDP-galactose transporter at the Golgi membrane. We suggest that the putative eighth (the most C-terminus-proximal) transmembrane helix of Gms1p is critical to targeting from ER to the Golgi membrane.

Amino Acid Sequence↗
PubMed2001

Characterization of a Schizosaccharomyces pombe mutant deficient in UDP-galactose transport activity.

In fission yeast, Schizosaccharomyces pombe, the carbohydrate components of the cell wall consist of galactomannan, unlike in Saccharomyces cerevisiae. We previously found that the disruption of gms1+, a gene encoding the UDP-galactose transporter required for the synthesis of galactomannan, led to the complete defect of cell surface galactosylation in Sz. pombe. The Deltagms1 strain is therefore useful for the analysis of physiological properties of galactose residues in Sz. pombe. The deletion strain of gms1+ was viable; however, itshowed an aberrant cell morphology and increased sensitivities to digestion with beta-glucanase and to various drugs, such as hygromycin B, sodium orthovanadate and Calcofluor white. A reduction of galactomannan layers of the cell wall in the Deltagms1 strain was observed by scanning and transmission electron microscopic analyses. The addition of osmotic stabilizer suppressed the morphologic defect of the Deltagms1 cells, while other phenotypes were weakly suppressed. The Deltagms1 (h90) strain was incapable of sexual conjugation during nutritional starvation. These results suggest that the cell surface galactosylation is required not only for non-sexual flocculation but also for sexual conjugation in Sz. pombe.

Biological Transport↗
PubMed2001

Somatic mutations of the PTEN/NMAC1 gene associated with frequent chromosomal loss detected using comparative genomic hybridization in endometrial cancer.

OBJECTIVES: We analyzed the mutational status of the transforming growth factor-beta type II receptor (TGF beta RII), BAX, and PTEN/MMAC1 genes as well as microsatellite instability (MI) in 29 consecutive cases of endometrial carcinoma operated on at the Cancer Institute Hospital (Tokyo, Japan). To identify chromosomal loss associated with significant somatic mutations, we conducted comparative genomic hybridization (CGH) analysis. METHODS: We conducted a direct sequence for mutational analysis of these genes. To examine copy number loss at the chromosomal regions bearing these genes, we used CGH analysis. CGH analysis may provides a genome-wide overview about tumor-associated genomic imbalances. RESULTS: Among nine tumors that showed the MI+ phenotype, four (44%) demonstrated a significant mutation with a definite amino acid change in the PTEN/MMAC1 gene. CGH analysis demonstrated that all four tumors (100%) showed chromosomal copy number loss around the locus of this gene, whereas four (57%) of seven tumors with PTEN/MMAC1 mutations showed chromosomal loss or double mutations in MI- carcinomas. The role of TGF beta RII and BAX genes is limited as a target gene of MI+ phenotype in endometrial cancer, because several mutations of these genes were detected but a chromosomal loss was demonstrated by CGH in only one tumor in MI+ endometrial cancers with mutation. CONCLUSIONS: This report reveals, by using CGH, that most MI+ endometrial cancers with PTEN/MMAC1 mutations as well as MI- tumors showed inactivation of both alleles of this gene, which strongly suggested the involvement of this gene in carcinogenesis.

Adenocarcinoma↗
PubMed2001

Crystallization and preliminary X-ray studies of V(1)-ATPase of Thermus thermophilus HB8 complexed with Mg-ADP.

Crystals have been grown of the V(1)-ATPase sector of the V-type ATP synthase complex (V(0)V(1)) from the thermophilic eubacterium Thermus thermophilus HB8. These crystals are grown by the vapor diffusion method in the presence of 5 mM Mg-ADP, from solutions containing 100 mM sodium acetate and 2 M sodium formate, pH 5.5. The crystals diffracted X rays beyond 3.4 A in resolution on a synchrotron radiation source. The crystals belong to the trigonal space group P3, with unit cell dimensions of a = b = 89.0 A, c = 179.2 A, and gamma = 120 degrees. The unit cell presumably contains one molecule of V(1)-ATPase and the V(m) value is calculated as 3.0 A(3)/Da.

Adenosine Diphosphate↗
PubMed2001

Safety evaluation of phosphodiesterase produced from Penicillium citrinum: summary of toxicological data.

The toxicity of Enzyme RP-1, an enzyme preparation used to hydrolyze yeast RNA to produce flavor enhancers for use in the food industry, was evaluated in a series of studies. A 5-week dietary toxicity study in Wistar rats was conducted in which animals received Enzyme RP-1 in feed at concentrations of 0, 500, 2000, or 8000 mg/kg body wt/day. A 13-week dietary toxicity study in Sprague-Dawley rats was conducted in which animals received RP-1 concentrate at 0, 0.125, 0.5, or 2% in their diets. At the highest dose levels in both studies, submandibular glands in the oral cavity were enlarged, an effect attributed to protease activity of the enzyme preparation. The no-observed-effect level in rats in the 13-week study was 0.5%, equivalent to 317 mg/kg body wt/day for males and 346 mg/kg body wt/day for females. Based on estimated dietary exposure to the enzyme preparation, the margin of exposure is estimated to be greater than 38,000. Lack of genotoxic potential was demonstrated by an in vitro reverse mutation assay in Salmonella typhimurium strains TA98, TA100, TA1535, and TA1537 and in Escherichia coli strain WP2uvr and by an in vitro chromosome aberration test in CHL/IU cells derived from fibroblasts from the lungs of Chinese hamsters. Finally, the particular strain of Penicillium citrinum, the fungal strain used to prepare Enzyme RP-1, was shown to have low pathogenicity upon a single injection into the tail vein of rats of viable spores at doses up to 2.8x10(5) colony-forming units per animal. The results of these studies demonstrate that the enzyme preparation may be considered safe to workers and consumers when employed in the production of flavor enhancers from yeast.

Administration, Oral↗
PubMed2001

A method for detecting myocardial abnormality by using a total current-vector calculated from ST-segment deviation of a magnetocardiogram signal.

A simple method to determine the state of ischaemia or fibrosis of myocardial cells has been developed. This method uses the ST wave of 64-channel magnetocardiogram (MCG) signals to calculate three parameters from the current-arrow map of the normal component signal of the MCG. One parameter is a total current vector that is obtained through summation of all current arrows. Another is a variance current vector calculated from the differential vector of two total current vectors at different times. The third is a flatness factor between the magnitude of the total current vector and the variance current vector. The three parameters are independent of the distance between the heart and the gradiometers. We measured the MCG signals of 29 healthy subjects, twenty patients with coronary artery disease (ten with previous myocardial infarction (MI) and ten with angina pectoris (AP)), and eight patients with cardiomyopathy (four with hypertrophic cardiomyopathy (HCM), three with dilated cardiomyopathy (DCM), and one with restrictive cardiomyopathy (RCM)). With our method, none of the healthy subjects tested positive for myocardial abnormalities, while 80% of the MI patients, 50% of the AP patients, and 100% of the cardiomyopathy patients tested positive. Although further testing is needed, we feel this simple technique enables easy diagnosis of myocardial damage.

Cardiomyopathies↗
PubMed2001

A method for detecting myocardial abnormality by using a current-ratio map calculated from an exercise-induced magnetocardiogram.

A method for making a current-ratio map to determine the ischaemic area of angina pectoris (AP) patients has been developed. This method uses a current-arrow map calculated using a ORS wave from 64-channel magnetocardiogram (MCG) signals. The current-ratio map can be calculated from the ratio of an exercise-induced current vector to an at-rest current vector. The MCG signals of eight patients with angina pectoris (AP) (six patients with effort AP and two patients with variant AP) and four healthy volunteers were measured before and after a two-step exercise test. The current-ratio maps of the six patients with effort AP showed three distinct patterns: a left-circumflex-artery (LCX) pattern; a right-coronary-artery (RCA) pattern; and a left-anterior-descending (LAD) pattern. The maximum current ratios of these three patterns differed from those of normal patterns. The patterns of two patients with variant AP were similar to normal patterns. Furthermore, a comparison of the current-ratio map before and after percutaneous-transluminal-coronary-angioplasty (PTCA) treatment indicated that the cardiac ischaemia was reduced in all patients. An appropriate criterion to diagnose abnormality in a patient with an ischaemic myocardial area seems to be a maximum current ratio exceeding 0.4 to 0.5. Based on these preliminary results, it is believed that the location of an ischaemic area (the coronary artery part) can be estimated by using the ischaemic current-ratio map pattern.

Angina Pectoris↗
PubMed2001

Changes in serotonin dynamics in the gastrointestinal tract of colon-26 tumour-bearing mice: effects of cisplatin treatment.

Severe nausea and vomiting are common side effects of anti-cancer chemotherapy. 5-HT3 receptor antagonists have been used for the treatment of these gastrointestinal symptoms. The purpose of this study was to examine whether specific changes in serotonin dynamics occurred in the gastrointestinal tract in mice in which Colon-26 adenocarcinoma cells were injected s.c., especially after treatment with cisplatin. The serotonin content of the small intestine of mice inoculated s.c. with Colon-26 adenocarcinoma increased significantly 2 weeks after the inoculation of the tumor cells; this was associated with an increase in tryptophan hydroxylase activity and the number of enterochromaffin cells as compared with control mice. Intravenous injection of cisplatin significantly reduced the serotonin content in the small intestine of Colon-26 tumour-bearing mice but not in control mice. The spontaneous release of serotonin from isolated intestine was not different between Colon-26 tumour-bearing and control mice; however, pretreatment of mice with cisplatin induced two fold increases in serotonin release from duodenum, jejunum and ileum in Colon-26 tumour-bearing mice but not in control mice. These results indicate that a region-specific increase in the number of enterochromaffin cells is observed in the intestine of Colon-26 tumour-bearing mice, associated with an increase in the serotonin content and tryptophan hydroxylase activity. Cisplatin treatment induced the release of serotonin from affected enterochromaffin cells in the gastrointestinal tract, which may be related to the occurrence of nausea in clinical use.

Animals↗
PubMed2001

Paired inspiratory-expiratory thin-section CT findings in patients with small airway disease.

The objective of this study was to evaluate minimal small airway disease (SAD) as reflected on paired inspiratory-expiratory CT findings. Seventy-two subjects, 34 with SAD, 11 with normal lung function, and 27 with chronic obstructive pulmonary diseases (COPD), underwent thin-section CT during deep inspiration and expiration at upper, middle, and lower lung levels. Evaluation of CT measurement was performed between inspiratory and expiratory CT attenuation of lung parenchyma, in non-dependent and dependent lung at each of the three levels. Visual assessment of mosaic patterns and attenuation differences was also performed using the inspiratory and expiratory images. Patients with SAD were differentiated from those with COPD, by visual assessment and from the CT measurements. Visual assessment failed to differentiate between the SAD and normal groups. However, one measurement, an inspiratory-expiratory attenuation difference in the dependent lower lung, was different between SAD and normal group. Early small airway disease may be indicated by an inspiratory-expiratory attenuation difference in the dependent lower lung using the simple method of a paired inspiratory-expiratory CT.

Adult↗
PubMed2001

Immunohistochemical detection of lipid peroxidation products, protein-bound acrolein and 4-hydroxynonenal protein adducts, in actinic elastosis of photodamaged skin.

Acrolein and 4-hydroxy-2-nonenal (HNE) are both byproducts of a lipid peroxidation reaction. Actinic elastosis in photodamaged skin of aged individuals is characterized by the accumulation of fragmented elastic fibers in the sun-exposed areas. To study whether a lipid peroxidation reaction is involved in the accumulation of altered elastic fibers in actinic elastosis, skin specimens obtained from sun-damaged areas were immunohistochemically examined using the antibodies against acrolein and HNE. Both antibodies were found to react with the accumulations of elastic material. Double immunofluorescence labeling demonstrated that acrolein/elastin and HNE/elastin were colocalized in the actinic elastosis. Western blot analysis showed that the polypeptide with a molecular weight of 62 kDa reacted with anti-acrolein, anti-HNE and anti-elastin antibodies. The results suggest that acrolein and HNE may be associated with actinic elastosis.

Acrolein↗
PubMed2001

Esophageal smooth muscle tumor in a 25-year-old woman with congenital malformations.

We recently treated a 25-year-old woman with an esophageal smooth muscle tumor with congenital malformations. Although the large size of the tumor and the presence of hemonecrotic lesion suggested the tumor to be leiomyosarcoma, histological studies revealed it to be leiomyoma. According to previous reports in the English-language literature, the coincidence of esophageal smooth muscle tumor with congenital malformations is relatively rare, and the coincidence of such a tumor with malformations of the type seen in this patient has never been reported. The congenital malformations in our patient were ocular hypertelorism, platyrrhiny, bilateral divergent strabismus, clubbed fingers and toes, fingerprint abnormality, and mild mental retardation. These congenital malformations cannot be explained by any reported syndromes.

Abnormalities, Multiple↗
PubMed2001

Gasless endoscopic thyroid and parathyroid surgery using a new retractor.

Endoscopic thyroid and parathyroid surgery have now become feasible procedures, but the working space provided by the gasless technique is more limited than that of the CO2 insufflation technique. Gasless endoscopic surgery was performed in 20 patients with thyroid or parathyroid tumors. A newly designed retractor was used. Gasless endoscopic surgery was performed in all patients without conversion to conventional techniques. The recurrent laryngeal nerve was visualized and preserved in all patients. No recurrent nerve palsy was noted. The new retractor created a sufficient working space, and our results demonstrated the feasibility of this technique.

Endoscopy↗