Search PubMed⌕ Search

Biomedical subjects

J Fujii

Publications and source records attributed to J Fujii.

At least 145 records · Page 8Linked to original sources
PubMed1994

Induction of manganese superoxide dismutase mRNA by okadaic acid and protein synthesis inhibitors.

We have reported that the phorbol ester phorbol 12-myristate 13-acetate (PMA) enhances the expression of manganese superoxide dismutase (Mn-SOD) mRNA [Fujii and Taniguchi (1991) J. Biol. Chem. 266, 23,142-23,146]. Okadaic acid, an inhibitor of type I and type IIa phosphatases, was also found to induce Mn-SOD mRNA at submicromolar concentrations in HeLa cells. Addition of cycloheximide resulted in superinduction of PMA- or tumour necrosis factor-stimulated expression of the mRNA, but not of okadaic acid-stimulated expression. When the effect of cycloheximide on the decay of Mn-SOD mRNA was examined by inhibiting mRNA synthesis with actinomycin D, cycloheximide had virtually no effect on mRNA stability, suggesting that accumulation of the mRNA was caused by activation by this reagent of transcription of the gene. PMA pretreatment of HeLa cells markedly enhanced cycloheximide-dependent superinduction of Mn-SOD mRNA. These data suggest that phosphorylation of several proteins is implicated in the regulation of Mn-SOD gene expression.

Animals↗
PubMed1994

Cerebellar infarctions secondary to cranio-cervical anomalies: a case report.

We report a case of cerebellar infarctions which occurred in the territories of the bilateral posterior inferior cerebellar arteries. This case was complicated with cranio-cervical anomalies composed of assimilation of the atlas, atlanto-axial dislocation, and basilar impression. The 40-year-old male patient had no detectable risk factors predisposing to atherosclerotic arterial occlusion or cardiogenic embolism, and there were no angiographic findings of atherosclerosis. It was, therefore, postulated that the cerebellar infarctions were secondary to those cranio-cervical anomalies. The developing mechanism is discussed.

Adult↗
PubMed1994

Suppression of antioxidative enzyme expression by transforming growth factor-beta 1 in rat hepatocytes.

We have investigated the effect of transforming growth factor-beta 1 (TGF-beta 1) and three cytokines on expression of antioxidative enzymes, manganese-superoxide dismutase, copper, zinc-superoxide dismutase, and catalase in cultured hepatocytes of rat. While interleukin-1 beta and interleukin-6 induced manganese-superoxide dismutase gene expression, they slightly suppressed catalase gene expression in rat hepatocytes. TGF-beta 1 suppressed expression of all these antioxidative enzymes in time- and cell density-dependent manners. Furthermore, we examined the effect of TGF-beta 1 on expression of glutathione peroxidase and glutathione-S-transferase, which exhibit glutathione-dependent peroxidase activity in rat hepatocytes. Expression of two major classes of the rat glutathione-S-transferase subunits 1 and 2 was also reduced by TGF-beta 1, although expression of glutathione peroxidase was not affected. Flow cytometric analysis indicated that production of peroxides was increased in hepatocytes treated with TGF-beta 1. These data suggest that augmented production of hydrogen peroxide and its intermediate through suppression of antioxidative enzyme expression may participate in cellular injury or growth inhibition promoted by TGF-beta 1.

Animals↗
PubMed1994

A novel mutation in Cu/Zn superoxide dismutase gene in Japanese familial amyotrophic lateral sclerosis.

Recently, several missense mutations in the Cu/Zn superoxide dismutase gene (SOD1) have been reported as a putative cause of chromosome-21q-linked familial amyotrophic lateral sclerosis (FALS). We have discovered a novel missense mutation (substitution of Thr for Ala4) in exon 1 (GCC to ACC) in two FALS patients from one Japanese FALS family. No mutations were found in 17 cases of sporadic ALS. The enzyme activity of recombinant fusion protein containing the Cu/Zn superoxide dismutase (SOD) with the Ala4-to-Thr mutation was significantly reduced in E. coli. On the other hand, in the expression system in insect cells using Baculovirus, the mutant SOD expressed an enzyme activity as high as wild-type SOD. These results suggest that the stability of SOD with the Ala4-to-Thr mutation is disrupted especially in the fusion protein. Autopsy was carried out on one of the two patients, and the pathological findings were typical of FALS with posterior column involvement. These results raise the possibility that mutation of the SOD1 is responsible for FALS with broader pathological involvement.

Amino Acid Sequence↗
PubMed1994

Nitric oxide synthase from rat colorectum: purification, peptide sequencing, partial PCR cloning, and immunohistochemistry.

Nitric oxide synthase (NOS) has been purified over 6,500-fold with a 3.4% yield from rat colorectum with 2',5'-ADP-Sepharose, DEAE cellulose, and gel filtration. The purified enzyme gave a single band corresponding to an apparent molecular mass of 160 Dka on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. When assayed in the requisite presence of L-arginine, CaCl2, NADPH, calmodulin, tetrahydro-L-biopterin, and FAD, the purified enzyme exhibited a specific activity of 328 nmol/min/mg L-citrulline formed and an apparent Km for L-arginine of 2.9 microM. Amino acid sequencing of 12 peptides revealed identical sequences to that of the neuronal type enzyme except for two altered amino acid residues. When partial reverse transcription-polymerase chain reaction of RNA from rat colorectum and cerebellum was performed using primers designed according to the amino acid sequences determined, these amino acid changes were found in both cDNA fragments, indicating the identity of the colorectal enzyme to the cerebellar one. A polyclonal antibody raised against NOS purified from rat cerebellum cross-reacted with the NOS from colorectum but not that from IFN-gamma stimulated macrophage-derived cells, RAW 264.7. Immunohistochemical analysis of the colorectum using this specific antibody indicated that Auerbach's plexus is strongly immunoreactive, supporting the hypothesis that NO is an inhibitory transmitter for non-adrenergic and non-cholinergic nerves in the colorectum.

Amino Acid Oxidoreductases↗
PubMed1994

Direct evidence of neuron impairment by oral infection with verotoxin-producing Escherichia coli O157:H- in mitomycin-treated mice.

We developed a mouse model of acute encephalopathy induced by verotoxin 2 variant (VT2v)-producing Escherichia coli. Three-week-old mice were inoculated intragastrically with approximately 10(10) CFU of E. coli O157:H- strain E32511/HSC and simultaneously given an intraperitoneal injection of mitomycin (MMC; 2.5 mg/kg). Drinking water containing 5 g of streptomycin sulfate per liter was given ad libitum from 3 days before the infection. From 1 to 2 days after bacterial inoculation, clinical features including weight loss, weakness, and flaccid paralysis of the extremities developed, usually culminating in death within 4 days. Diarrhea was not observed during the course of disease. No mice died in the absence of streptomycin or MMC treatment for 2 weeks after the oral bacterial infection. Judging from the clinical course and the biochemical and histological examination, the cause of death was not likely to be attributable to renal failure or to a side effect of MMC. To better understand the cause of death, we examined the brain cortex and spinal cord of the moribund mice by electron microscopy. Mice showing mortal symptoms were given horseradish peroxidase intravenously. The tracer was present in the endothelial basal lamina, in the surrounding extracellular spaces, and even in the neuron fibers of the brain cortex. Furthermore, immunoreactivity of VT2v, proved by the use of rabbit anti-VT2 serum, was localized selectively in the damaged myelin sheaths of neuron fibers which were accompanied by edematous axons in the brain cortex and spinal cord. These findings strongly suggest that VT2v is toxic to both endothelial cells and neurons in the central nervous system and subsequently causes fatal acute encephalopathy.

Acute Disease↗
PubMed1994

[Reference values of laboratory tests in elderly subjects--blood pressure].

The upper limits of normal blood pressure have been considered to be 139 mmHg systolic and 89 mmHg diastolic for adults, but these values are not necessarily applicable to the elderly. This report presents blood pressure values of healthy persons aged 65 to 94 and estimates the upper limits of normal blood pressure in the elderly based on follow-up studies. The Blood Pressure Subgroup of the Study on Reference Values of Laboratory Tests in Elderly Subjects defined inclusion criteria for the healthy elderly as follows: (1) persons aged 65 to 94, (2) persons not complicated with cardiovascular diseases, (3) persons capable of living and walking freely, (4) persons without dementia, (5) persons without anemia, liver disease, renal failure, diabetes mellitus on drug treatment, lung disease, valvular disease or marked arrhythmias, (6) persons without neuromotor disease. The subgroup collected 2008 persons who fulfilled the criteria. Of the 2008 persons, 663 were not taking antihypertensive drugs, had body weight within an average Body Mass Index +/- standard deviation and had no abnormalities on ECG. The 663 persons were considered to be a group of most the normal elderly. Blood pressure values in this group were 133.3 +/- 18.9/77.0 +/- 10.6 mmHg for males (N = 318) and 134.3 +/- 18.7/75.7 +/- 10.2 mmHg for females (N = 345). Follow-up studies carried out by some members of the Blood Pressure Subgroup suggested that the upper limits of the normal blood pressure were 140 to 159 mmHg systolic and 80 to 89 mmHg diastolic for the elderly.

Adult↗
PubMed1994

[Echocardiographic assessment of cardiac basal hypertrophy represented by increased base to mid thickness ratio (B/M ratio) in the ventricular septum and its relation to age].

The basal part of the interventricular septum may easily become hypertrophic because it is exposed to strong hemodynamic stress compared to the other portions of the left ventricle. We measured the end-diastolic interventricular wall thickness both at the base and in the midsection by 2D echocardiography in 122 normotensives, and examined whether the basal thickness increases with age. The basal thickness (B) increased with age in both sexes. In males the thickness averaged 10.1 mm in the 50-59 age group, 10.2 m in those aged 60-69 and 11.4 mm (p < 0.01) in those 70 or older compared to 9.4 mm in those aged 49 or younger. In females it was 8.1 mm (p < 0.05) in the 50-59 age group, 8.3 mm (p < 0.05) in those aged 60-69 and 10.0 mm (p < 0.01) in those 70 or older compared to 6.8 mm in those 49 or younger. Concerning the midwall thickness (M), there were no significant changes among the respective age groups in either sex. As a result, a close correlation was found between the B/M ratio, a new and simple index for basal hypertrophy, and age (R = 0.46, p < 0.01 in males and R = 0.43, p < 0.01 in females). Comparison of the B/M ratio between the two age groups 49 or younger and 70 or older was as follows; 1.08 vs 1.30 (p < 0.01) in males and 1.01 vs 1.27 (p < 0.01) in females. Increase of basal hypertrophy in the aged was clearly indicated by the B/M ratio.

Adult↗
PubMed1994

[Patient-hospital relationship and quality of life in elderly patients with hypertension].

To investigate the influence of the patient-hospital relationship on the quality of life (QOL) in elderly patients with hypertension, we developed a comprehensive scale for assessing the patient-hospital relationship. In the first phase, an original questionnaire consisting of 40 items was designed under five fields for global assessment of the concept. We administered this questionnaire to 715 patients with cardiovascular diseases. Variations in the scores were evaluated by factor analysis. Three factors that comprised the majority of variance were derived by principal components analysis and varimax transformation. These orthogonal factors were considered to reflect "convenience of the outpatient clinic", "satisfaction with clinical staff", and "communication with physician". We adopted three items under each of the three domains. Using this instrument, we assessed the patient-hospital relationship in 402 elderly outpatients with hypertension at 8 clinical centers. QOL was simultaneously assessed. A linear regression analysis revealed a significant positive correlation between patient-hospital relationship and QOL scores (R = 0.403, p < 0.0001), which means that patient-hospital relationship explained up to 16 percent of the total variability in scores on the Overall Quality of Life scale. Closer correlations were observed in patients aged 80 years or older, and in patients who lived alone. These results indicate that the patient-hospital relationship is one of the substantial factors determining QOL in elderly hypertensive patients, with more substantiality in older patients and patients leading solitary lives.

Aged↗
PubMed1994

[The association between body mass index and self-assessed symptoms among young women].

Attitudes and behaviours of young women toward dieting and the relationship between body mass index (BMI) and subjective symptoms were studied by a questionnaire survey applied to all the students of a two-year women's college in Sapporo, Hokkaido. The response rate was 96% to yield a total of 608 subjects. 1) Young women showed excessive thinness-oriented attitudes. 2) Over 30 percent of the women reported irregular menstruation, neck pain, fatigue and oversensitiveness to cold. 3) Those who reported oversensitiveness to cold, wearied eyes or stomachache had significantly lower average BMI than those who did not. 4) A U-shaped relationship, with a higher prevalence of complaints of these three symptoms with a low BMI, was shown. Considering these findings, it appears that being excessively thin is related to symptoms and young women's thinness-oriented attitudes are unhealthful.

Adolescent↗
PubMed1993

Induction and release of manganese superoxide dismutase from mitochondria of human umbilical vein endothelial cells by tumor necrosis factor-alpha and interleukin-1 alpha.

The effects of tumor necrosis factor-alpha (TNF alpha) on cultured human umbilical vein endothelial cells (EC) and 2 cancer cell lines, A549 and ME180, were compared. The effects of interleukin-1 alpha (IL-1 alpha) on EC were also examined. While A549 cells were fairly resistant to the cytolytic effects of TNF alpha and IL-1 alpha, ME180 cells were sensitive. EC were also less sensitive to TNF alpha than ME180 cells, as judged by viability of individual cells and by the release of lactate dehydrogenase (LDH) into the medium. Both manganese superoxide dismutase (Mn-SOD) and its mRNA were markedly induced by these cytokines in EC and in A549 cells but not in ME180 cells. The levels of Mn-SOD in the conditioned medium of EC were markedly increased after stimulation with cytokines, whereas those in ME180 and A549 cells were relatively low. The amount of Mn-SOD released appears to be comparable to that from cells lysed due to the cytocidal effect of cytokines, as assessed by measuring intra- and extra-cellular LDH activity. These data suggest that, in vivo, the TNF alpha and IL-1 alpha produced by cancer cells and other cells may induce Mn-SOD in vascular endothelial cells as well as other host tissues, resulting in release of a relatively large amount of this protein into the serum.

Blotting, Northern↗
PubMed1993

Identity of a major 3-deoxyglucosone-reducing enzyme with aldehyde reductase in rat liver established by amino acid sequencing and cDNA expression.

We have purified a rat liver enzyme that catalyzes the NADPH-dependent reduction of 3-deoxyglucosone (3-DG), a major intermediate in the Maillard reaction and a potent cross-linker responsible for the polymerization of proteins. Comparison of the amino acid (aa) sequences of nine peptides obtained from the rat 3-DG-reducing enzyme by lysylendopeptidase digestion with the aa sequence of human aldehyde reductase (ALR) [Bohren et al., J. Biol. Chem. 266 (1991) 24031-24037] strongly suggested that the purified enzyme was rat ALR. We cloned the cDNA encoding ALR from a rat kidney cDNA library using a human ALR cDNA fragment, amplified by polymerase chain reaction, as a probe. All nine peptides identified in the purified rat 3-DG-reducing enzyme were found in the aa sequence deduced from the rat ALR cDNA. Moreover, cell extract from COS-1 cells transfected with the rat ALR cDNA exhibited NADPH-dependent 3-DG-reducing activity and cross-reacted with antiserum raised against the purified rat 3-DG-reducing enzyme. All the above data indicate clearly that the 3-DG-reducing enzyme is identical with ALR. Northern blot analysis of total mRNA from a variety of rat tissues showed fairly high levels of expression of ALR mRNA. This suggests that sufficient ALR is present to detoxify 3-DG when it is formed through the Maillard reaction in vivo.

Aldehyde Reductase↗
PubMed1993

Significance of Arg-107 and Glu-108 in the catalytic mechanism of human gamma-glutamyl transpeptidase. Identification by site-directed mutagenesis.

The catalytic properties of wild-type and mutant enzymes produced by expression of cDNA (for gamma-glutamyl transpeptidase) in COS-1 cells were analyzed. Substitutions of Lys-100 to Asn, Glu-102 to Gln, and ARg-112, -139, -147, and -150 to Gln resulted in expression of fully active enzymes. Replacement of Arg-107 with Gln or His led to complete loss of enzyme activity, whereas substitution to Lys resulted in partial loss of activity, indicating that the positive charge at position 107 is essential for enzyme activity. In the Arg-107-to-Lys mutant, the apparent Km for the donor L-gamma-glutamyl-3-carboxy-4-nitroanilide was increased 8.4-fold compared to that of the wild-type enzyme. Although 32% of the wild-type hydrolysis activity was retained in this mutant, the activity could not be enhanced by addition of an acceptor substrate, glycylglycine, indicating that the transpeptidation reaction was specifically prevented. Substitution of Glu-108 to Gln increased the apparent Km for glycylglycine 8.5-fold while increasing the apparent Km for the donor only 1.6-fold. The apparent Vmax was decreased to 4-5% of wild-type for both substrates. These data suggest that Arg-107 plays a significant role in substrate binding rather than catalysis, and that Glu-108 is an important, although not essential, participant in both acceptor binding and catalysis.

Amino Acid Sequence↗
PubMed1993

Chromosome mapping of five human cardiac and skeletal muscle sarcoplasmic reticulum protein genes.

Fluorescence in situ hybridization (FISH) experiments were performed using genomic and complementary DNA probes in order to determine the location on human chromosomes for five genes expressed in cardiac and skeletal muscle sarcoplasmic reticulum. The chromosome location of each gene was determined in terms of both cytogenetic bands and fractional chromosome length. The ATP2A2 gene, expressing the SERCA2 isoform of the Ca2+ pump, maps to bands 12q23-q24.1, the phospholamban gene (PLN) to 6q22.1, the human skeletal muscle calsequestrin gene (CASQ1) to band 1q21, the cardiac calsequestrin gene (CASQ2) to bands 1p11-p13.3, and the cardiac calcium release channel gene (RYR2) to the interval between band 1q42.1 (distal) and band 1q43 (proximal).

Adenosine Triphosphatases↗
PubMed1993

A new model for infective endocarditis of the mitral valve in rabbits.

To produce an experimental model of infective endocarditis without inserting catheters into the heart, we injected a bacterial suspension into rabbits in which mitral complex lesions had been induced by electrical stimulation of the cervical vagus. Typical infective vegetations grew on the surface of the mitral valves 1 week later. The formation of vegetations was related to the timing of the inoculation. Streptococcus viridans injected just after vagal stimulation produced vegetations in 10 of 17 animals (58.8%), but the same bacteria injected 14 days after vagal stimulation did so in only 2 of 11 animals (18.2%). The incidence of infective endocarditis was significantly higher after early inoculation compared with delayed inoculation (p < 0.01). The susceptibility to infection depended on the species of bacteria injected. Both S. viridans and Pseudomonas pseudoalkaligenes injected just after vagal stimulation produced vegetations in 10 of 17 (58.8%) and 6 of 13 (46.2%) animals, respectively, but Staphylococcus epidermidis injected just after vagal stimulation did not produce vegetations in any of the 10 animals. S. viridans injected into nine normal animals never produced vegetations. These findings indicate that infective endocarditis develops after intravenous injection of bacterial suspensions alone in rabbits with mitral complex lesions.

Animals↗
PubMed1993

Expression of DNA methyltransferase in LEC rats during hepatocarcinogenesis.

The Long-Evans with a cinnamon-like color (LEC) rat is a mutant of the Long-Evans strain that develops hereditary hepatitis and hepatoma with ageing. Age-related changes in the mRNA expression of DNA methyltransferase (DNA MTase) were examined in livers of LEC rats using Long Evans with an agouti color (LEA) rats as controls. A dramatic increase in the expression of this mRNA was observed in LEC rats at 20 weeks when acute hepatitis appeared. Their high mRNA levels were maintained until 52 weeks of age. The mRNA expression as well as DNA MTase activities were found to be higher in cancer lesions than in adjacent normal tissue. These increases may be related to liver regeneration and to early events in cellular transformation of LEC rats.

Aging↗