[ADAM family proteins and cell fusion].
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Biomedical subjects
Publications and source records attributed to T Endo.
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The diet of 686 medical student at Hirosaki University were studies between 1986 to 1993. Food groups that showed a higher intake in males than in females were grains, meat, liquor, and others. On the other hand, the food groups that showed a higher intake in females were confectioneries, fruits, green vegetables, mushrooms, and seaweed. The nutrients that showed a higher intake in males than in females were energy, protein, fat, carbohydrates, and vitamins B1 and B2. On the other hand, only vitamin C showed a higher intake in females. Mean daily dietary fiber intake per student was 19.4 g by the Southgate method and 15.9 g by the Prosky method in males and 18.3 g by the Southgate method and 16.9 g by the Prosky method in females. These values were lower than the mean value for Japanese adults by about 4 g. Overall the intake from grains was disproportionately higher in males compared to females. Energy and nutrient intakes met recommended dietary allowance except for energy in females which was lower (93.7%).
It has been well known that tuberculous meningitis cases have moderate pleocytosis with mononuclears as predominating cell type in cerebrospinal fluid (CSF). But there are some reports of tuberculous meningitis in which polymorphonuclears was predominant in CSF. In this study, we investigated differential cell counts of CSF in 4 tuberculous meningitis cases. In 3 of the 4 cases, the CSF had a polymorphonuclears preponderance on admission. The elevated ratio of polymorphonuclears seemed to relate with the severity of the disease, and the ratio declined promptly after the initiation of chemotherapy. The 2 cases, which showed extremely high ratio of polymorphonuclears, showed poor prognosis, one died and the other had the severe sequelae such as notable disturbances of consciousness with hydrocephalus. This study suggests that differential cell counts of CSF is a useful measures to predict the prognosis and to follow up patients with tuberculous meningitis. The finding of a polymorphonuclears preponderance in CSF would suggest the severity and activities of the disease and should be considered as an alarm sign in tuberculous meningitis.
In order to examine the use of helical computed tomography (CT) in lung cancer screening, we evaluated the effects of tube current and table feed speed on image quality. CT images of normal volunteers, which were scanned under various tube currents (150mA, 100mA, 50mA) and various table feed speeds (15mm/sec, 20mm/sec, 25mm/sec, 30mm/sec), were visually evaluated by experienced diagnostic radiologists. The images were not affected by decreasing the milliamperage. However, as table feed speed increased, images were evaluated as significantly worse. Particularly, since the degradation of the images scanned at greater than 25mm/sec was remarkable, they were judged to be unacceptable for detection of the abnormality. In conclusion, a tube current of less than 50mA and table feed speed of less than 20mm/sec are suitable in the application of helical CT for lung cancer screening.
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Peripheral mononuclear cells from 47 patients suffering from Japanese cedar pollinosis were stimulated with crude pollen extract of Cryptomeria Japonica (CJ) to evaluate the effect of CJ-specific immunotherapy. The stimulation index (SI) with crude pollen extract of CJ in the patients with excellent and good clinical outcome was significantly lower than those with poor outcome. The SI increased with the increasing symptoms in a dose dependent manner. In the patients under immunotherapy for long period, the good clinical outcome and low SI were gained. These results suggest that peripheral lymphocyte proliferation test should be a good objective indicator for allergen-specific immunotherapy.
The activities of cell-associated IL-1 (IL-1 alpha) and extracellular IL-1 (IL-1 beta) in alveolar macrophages (AM) from rats with bleomycin-induced pulmonary fibrolosis were measured to determine their role in fibroblast growth. AM were obtained on Days 1, 3, 6, 9 and 12 after a intratracheal injection of bleomycin, and the IL-1 activity and fibroblast growth-stimulating activity in fixed AM and AM supernatants were measured. Higher cell-associated IL-1 activity was detected in AM from bleomycin-treated rats than in those of control on Day 1 through 9. But extracellular IL-1 activity in the supernatant of AM from bleomycin-treated rats significantly higher only on Day 1. Expression of IL-1 alpha mRNA in AM from bleomycin-treated rats was significantly higher than that in AM of control, but there was no significant difference in the mRNA levels of IL-1 beta in AM of these two groups. Fixed AM from bleomycin-treated rats caused growth-inhibition of fibroblasts in a density-dependent manner. The inhibitory activity was decreased by pretreatment of fixed AM with anti IL-1 alpha antibody, but not anti IL-1 beta antibody. These results suggest that cell-associated IL-1 (IL-1 alpha) is produced continuously in AM from rats with bleomycin-induced pulmonary fibrosis and may be important in regulation of this disorder by inhibiting fibroblast growth.
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MAS17 (MAS22) is an essential component of the import receptor complex in the yeast mitochondrial outer membrane. MAS17 consists of three distinct domains: the N-terminal cytosolic domain, the internal membrane-spanning domain, and the C-terminal intermembrane space domain. In the present study, we examined the roles of the C-terminal domain of MAS17, which is rich in acidic amino acids, in protein import into mitochondria both in vivo and in vitro. Cells expressing MAS17 delta 120-152, a mutant MAS17 lacking the C-terminal acidic domain, could grow as fast as those expressing wild-type MAS17, while cells expressing MAS17 delta 97-152, a mutant MAS17 lacking both the intermembrane space and the membrane-spanning domains, stopped growing as soon as wild-type MAS17 was depleted. MAS17 delta 120-152 was correctly integrated into the mitochondrial outer membrane like wild-type MAS17. Mitochondria containing MAS17 delta 120-152 instead of wild-type MAS17 could import both authentic and artificial mitochondrial precursor proteins nearly as efficiently as wild-type mitochondria in vitro. These results suggest that the C-terminal intermembrane space domain of MAS17 is not essential for targeting or functions of MAS17.
An interesting case of acute poisoning by chromate compounds is reported. A 51-year-old man committed suicide by ingesting a fatal dose of sodium chromate solution. He unexpectedly lost consciousness 6 h after the ingestion and died approximately 20.5 h later. An examination of the blood showed noticeable hepatic damage and thrombocytopenia. The postmortem examination revealed extensive bleeding in the alimentary tract and a severe hepatic lesion due to hepatocellular necrosis. However, the renal disorder was unusually light in the microscopic and clinical findings. Moreover, the renal lesion was observed mainly in the distal tubules instead of the proximal tubules which is more typical in cases of acute poisonings by diverse heavy metals including chromium. The patient's death was assumed to have been caused by circulatory collapse due to internal bleeding and the direct toxicity of chromate compounds with hepatic malfunction and possibly disseminated intravascular coagulation (DIC).
We have developed a culture system of guinea pig tracheal epithelial cells using the epithelium-denuded human amnion as a source of basement membrane. Culture medium fluid over the epithelial cells was replaced by air after 7-day immersion culture and thereafter maintained for 2 weeks. Electron microscopical observations revealed that the height of epithelial cells and the ratio of ciliated epithelial cells looked like that of normal guinea pigs epithelium growth in 2 weeks after the air interface, but that no goblet cells could be found. In order to study cell polarity, we measured endothelin-1 levels in the media of apical and basal sides of the epithelial cell monolayer by means of the enzyme-linked immunosorbent assay. The endothelin-1 content of the submucosal side was over 30 times higher than that of the apical side. These findings suggest that ET-1 would be mainly released from airway epithelial cells toward the submucosal side.
We report here the isolation of the cDNA for pea chloroplast SecA. Pea SecA encodes a polypeptide of 1,011 amino acids and shows high sequence similarity with cyanobacterial SecA. Pea SecA was synthesized as a larger precursor and was imported into isolated chloroplasts in vitro. The purified pea SecA, which was expressed in Escherichia coli cells, stimulated the in vitro import of the 33 kDa protein of the oxygen-evolving complex into thylakoids. These results indicate that higher plant chloroplasts contain a bacterial-type SecA protein-dependent system for the intraorganellar protein transport into thylakoids.
Thyrotropin receptor (TSH-R) has been thought to be thyroid-specific, but, by Northern blot analysis, we found that rat adipose tissue expressed TSH-R mRNAs in amounts approaching those in the thyroid. To investigate the function of TSH-R from adipose tissue, we screened a rat fat cell lambda gt11 cDNA library for TSH-R sequences using a 32P-labeled rat thyroid TSH-R cDNA as a probe. Among 10(6) plaques, we obtained four positive clones. Sequencing of these cDNAs has revealed that two of them (F alpha and F beta) contained both initiation and termination codons. Comparison of F alpha with the thyroid TSH-R cDNA sequence revealed that F alpha was almost identical to the thyroid TSH-R, except that nucleotides 1041 and 1277 were changed from A to G and from C to T, respectively. In contrast, we found that F beta contained 21 novel nucleotides between nucleotides 467 and 468 of the thyroid TSH-R cDNA, encoding an additional 7 amino acids. However, when we prepared mRNA from adipose tissue and transcribed it into cDNA, we failed to amplify the F beta type of TSH-R cDNA by polymerase chain reaction, suggesting that F beta mRNAs are rare in the tissue. We then ligated F cDNAs into pSG5 and transfected them with pSV2-neo into Chinese hamster ovary (CHO)-K1 cells. TSH stimulated cAMP formation in CHO-F alpha cells in a manner similar to that in CHO cells transfected with thyroid TSH-R cDNA. In contrast, no increase of cAMP was observed in CHO-F beta cells. IgG from patients with Graves' disease (n = 4) showed thyroid-stimulating antibody activity only in CHO-F alpha cells (1288-4582%). In addition, CHO-F alpha cells and CHO cells transfected with thyroid TSH-R showed similar 125I-TSH binding activity. These results indicate that the fat cell expresses high levels of a TSH-R whose function is indistinguishable from that in the thyroid and suggest that the TSH-R autoantibody plays an important role in the pathogenesis of the extrathyroidal manifestations of Graves' disease.
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UDP-6-Deoxy-D-galactose and UDP-6-deoxy-6-fluoro-D-galactose were synthesized and their transfer to 2-acetamido-2-deoxy-D-glucose (N-acetyl-D-glucosamine) by beta-(1-->4)-galactosyltransferase was examined. The transfer rates of 6-deoxy-D-galactose and 6-deoxy-6-fluoro-D-galactose were 1.3 and 0.2% of that of D-galactosyl transfer, respectively. The 2-acetamido-4-O-(6-deoxy-beta-D-galactopyranosyl)-2-deoxy-D-glucopyranose (6'-deoxy-N-acetyllactosamine) and methyl 2-acetamido-4-O-(6-deoxy-6-fluoro-beta-D-galactopyranosyl)-2-deoxy-D- glucopyranoside (6'-deoxy-6'-fluoro-N-acetyllactosamine) were synthesized enzymatically in 30 and 59% yields, respectively. Further, 6-deoxy-D-galactose could be completely transferred to N-linked type biantennary oligosaccharides having two N-acetyl-D-glucosaminyl residues at the nonreducing end to give the corresponding penta- and hepta-saccharides in 55 and 57% yields, respectively. An assay of 6-deoxy-D-galactosyl transfer using asialo agalacto alpha 1-acid glycoprotein as an acceptor suggested that 6-deoxy-D-galactose was transferred to about 30% of the N-acetyl-D-glucosaminyl residues in the N-linked oligosaccharides of the glycoprotein.
The mouse myoblast cell line C2C12 constitutively expressed 160-kDa transmembrane NCAM isoform and 135-kDa GPI-anchored isoform before differentiation. During differentiation into multinucleated myotubes, the cells newly expressed 150-kDa GPI-anchored isoform and the level of 135-kDa GPI-anchored isoform increased. Structural analysis of the GPI glycan of NCAM, which was purified from C2C12 myotubes after metabolic labeling with [3H]inositol, was performed by sequential exoglycosidase digestion and Wistaria floribunda agglutinin-agarose column chromatography. The core GPI glycan structure, Man alpha 1-2Man alpha-Man alpha-GlcNH2-myoInositol, was conserved and variations were observed in additional mannose and N-acetylgalactosamine residues. Structural analysis of the GPI glycans of the two GPI-anchored isoforms, GPI-NCAM 135 and GPI-NCAM 150, showed the enhanced attachment of the N-acetylgalactosamine residue to the GPI glycan core of GPI-NCAM 150. These GPI-anchored NCAM isoforms were released from C2C12 cells during the myoblast differentiation. Release of GPI-anchored NCAMs was observed when C2C12 cells were cultured in a serum-free medium, and inositol but not inositol phosphate was detected after nitrous acid deamination of the released NCAM. These results suggest that the GPI-anchored NCAM was released from the cell surface by the action of an endogeneous phospholipase D.
Previous work has suggested that the antinociceptive effect of nitrous oxide (N2O) in rats is mediated, at least in part, by beta-endorphin (beta-EP) and that centrally administered beta-EP stimulates release of methionine-enkephalin (ME) in the rat spinal cord. Since inhibition of central nitric oxide (NO) production has been found to suppress N2O antinociception, we examined the possible involvement of NO in the release of spinal cord ME by i.c.v. beta-EP. Urethane-anesthetized, male Sprague-Dawley rats were intrathecally (i.t.) perfused with artificial cerebrospinal fluid (aCSF) and fractions of perfusate were assayed for immunoreactive (i.r.) ME. The beta-EP-induced increase in ME concentration in the i.t. perfusate was significantly suppressed by perfusing the animal with aCSF containing 100 microM L-NG-nitro arginine (L-NOARG), an inhibitor of NO synthase (NOS). The further addition of 50 microM L-arginine (L-ARG), but not D-arginine (D-ARG), to the aCSF reversed the suppression of the ME change by L-NOARG. However, the potency of L-ARG decreased with increasing concentrations of L-ARG. On the other hand, increasing the concentration of L-NOARG in the aCSF to 250 microM failed to produce a greater suppression of the beta-EP-induced increase in ME. These findings suggest that NO may mediate the beta-EP-induced release of ME in the spinal cord and that interference with this mechanism might be an explanation for the antagonism of N2O antinociception in rats by NOS inhibitors.
Cyclin-dependent kinase 2 (Cdk2) controls the transition from the G1 to the S phase in the mammalian cell cycle. We found by immunoblotting that anti-Cdk2 antibodies recognize three Cdk2 proteins (of 33, 34 and 39 kDa) in FRTL-5 and FRTL-Tc cells (malignantly transformed FRTL cells). Although 33 kDa protein is a phosphorylated form of 34 kDa protein previously reported, the nature of 39 kDa protein is unknown. In order to determine the nature of this protein, we screened a FRTL-5 cDNA library. Two cDNA clones of the rat homologue (rat Cdk2-alpha and -beta) of human Cdk2 were isolated. The open reading frame of rat Cdk2-alpha cDNA encoded a protein with 428 amino acids and has a high degree of conservation with human Cdk2. The calculated molecular weight of Cdk2-alpha protein is 33892 Da. The rat Cdk2-beta cDNA was identical to Cdk2-alpha cDNA except that it had extra 144 bp; this coincided with insertion of 48 amino acids into Cdk2-alpha protein between Met 196 and Val 197. The calculated molecular weight of Cdk2-beta protein is 39087 Da. Northern blot analysis indicated that the sizes of rat Cdk2-alpha and -beta mRNAs are approximately 2.5 kb and 3.0 kb, respectively. Partial proteolytic mapping showed that Cdk2-beta gene product is 39 kDa Cdk2 in the immunoblotting. We also found that Cdk2-beta protein binds to cyclin A and suc1 proteins. During G1-S phase in FRTL-Tc cells, Cdk2-alpha protein level is constant, but is gradually phosphorylated. In contrast, the level of Cdk2-beta protein increases through the S phase and decreases at the early G2 phase. These results suggest that a variant form of Cdk2 protein might be required for entry into the S phase of the cell cycle in FRTL-Tc cells.