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E Ueda

Publications and source records attributed to E Ueda.

At least 55 records · Page 3Linked to original sources
PubMed1995

Chromosomal loci of 50 human keratinocyte cDNAs assigned by fluorescence in situ hybridization.

The chromosomal loci of expressed genes provide useful information for a candidate gene approach to the genes responsible for genetic diseases. A large set of randomly isolated cDNAs catalogued by partial sequencing can serve as a resource for accessing and isolating these disease genes. Using fluorescence in situ hybridization, we examined the chromosomal loci of 217 human keratinocyte-derived cDNAs, with independent novel sequence tags at the 3' end region. Among them, we determined the loci of 50 cDNAs. Single-pass sequencing of these from the 5' ends indicated that 39 cDNAs still can be produced for new genes. These cDNAs with identified chromosomal loci are powerful tools that can be used to help elucidate the genes responsible for hereditary skin disorders.

Chromosome Mapping↗
PubMed1995

Chimeric horse/human recombinant C9 proteins identify the amino acid sequence in horse C9 responsible for restriction of hemolysis.

Equine C9, in contrast to human C9, has extremely low hemolytic activity against most mammalian erythrocytes, although the amino acid sequences of both proteins show 77% identity. In an attempt to define the region of human C9 responsible for conferring its lytic activity, or conversely, the region of equine C9 responsible for its restriction, recombinant human and equine C9 and four chimeric human/equine C9 proteins were constructed and expressed in COS-7 cells. Recombinant human and equine C9 displayed hemolytic profiles similar to those of the purified native proteins. Exchange of a fragment extending from residues 145 to 290 in horse C9 with the corresponding one from human C9 created a fully hemolytic protein. This region contains the putative hinge region but not the membrane-interacting domain. Nonlytic chimeric C9 proteins inhibited hemolysis and binding of human C9 to EAC1-8 cells, indicating that they bind to their receptor, but subsequent unfolding or insertion into the membrane is impaired. These results suggest that restriction factors, such as glycophorin, CD59, or homologous restriction factor, on erythrocytes may limit the activity of horse C9 by interacting with its hinge region. In support of this conclusion direct binding of CD59 to immobilized horse C9 was detected by ligand blotting, and it was observed that a polyclonal anti-CD59 Ab enhanced human and horse C9-mediated hemolysis of human EAC1-7, but the increase in hemolytic activity of horse C9 by inhibition of CD59 was less than what could be achieved by insertion of the human C9 hinge region into horse C9.

Amino Acid Sequence↗
PubMed1995

Plasma N-methylhistamine concentration as an indicator of histamine release by intravenous d-tubocurarine in humans: preliminary study in five patients by radioimmunoassay kits.

Histamine disappears rapidly from plasma because of its short half-life. Because a metabolite of histamine, N-methylhistamine, is stable and has a longer half-life, determination of its plasma concentration can be useful in retrospective determination of histamine release. In this study, we measured plasma histamine and N-methylhistamine concentrations after d-tubocurarine (dTc) administration to evaluate the use of plasma N-methylhistamine measurement for confirming histamine release. After the induction of anesthesia, five patients received dTc, 0.8 mg/kg intravenously. A radioimmunoassay kit was used to determine plasma histamine and N-methylhistamine before and 1, 3, 5, and 10 min after administration of dTc. Histamine released by the injection of dTc reached a maximum level at 1 min, but decreased rapidly, whereas N-methylhistamine increased at 1 min and remained increased for at least 10 min. Good correlations were found between histamine concentration at 1 min and N-methylhistamine concentrations at 1, 3, 5, and 10 min, especially r = 0.999 (n = 5) at 10 min. N-Methylhistamine measurement with this kit can ascertain histamine release retrospectively in a semiquantitative manner.

Anesthesia↗
PubMed1995

Statistical survey from 1982 to 1991 of 49 patients with malignant melanocytic tumors.

We investigated the clinical characteristics and outcome of 49 patients with malignant melanoma: 17 with acral lentiginous melanoma; 14 with nodular melanoma, six with superficial spreading melanoma, one with lentigo maligna melanoma, eight with melanoma in situ, one with malignant blue nevus, and two with melanoma of unknown origin. Of the 41 patients without melanoma in situ, 34.1% were in stage I, 17.1% in stage II, and 48.8% in stage III. No patients had reached stage IV. All patients with stage I, II, and III melanoma were treated with wide resection, lymph node dissection including prophylactic dissection, and combination chemotherapy with dacarbazine, nimustine hydrochloride, and vincristine (DAV) with or without Interferon-beta. The statistical analysis revealed that tumor thickness and level of invasion were factors significantly associated with outcome. A gradual increase in the number of new cases of melanoma was seen each year of the registration period (1982-1991); there was an approximately 4-fold increase during this decade over the previous decade. The majority of the primary lesions (80.5%) were located on the upper or lower extremities, the pT4 tumor thickness subgroup was the most frequent (39.0% of all melanomas), and invasion level IV was the most common (42.5%). About half (51.2%) of the melanoma patients were stage I or II; this group had a 5-year survival rate of 100%. The stage III patients had a 5-year survival rate of only 54.2% (p < 0.05).

Adolescent↗
PubMed1994

Cataloging of the genes expressed in human keratinocytes: analysis of 607 randomly isolated cDNA sequences.

The partial nucleotide sequences of 607 cDNAs randomly isolated from a cDNA library of cultured human epidermal keratinocytes were determined by single pass sequencing. Homology search of the sequences to the non-redundant nucleotide databases revealed that 27% of the cDNAs matched registered human-or non-human genes encoding not only keratinocyte specific genes, but also a variety of functional proteins, the expression of which had not been identified in keratinocytes. Non-matching cDNAs covering 49% of the cDNAs were not homologous even to ESTs from other organs, suggesting that these cDNAs include novel genes expressed in the cells. The large scale sequencing of keratinocyte cDNAs provides a useful molecular source for research into biology and diseases of the skin.

Animals↗
PubMed1994

Pulmonary hyalinizing granuloma with Castleman's disease.

A case of pulmonary hyalinizing granuloma (PHG) with Castleman's disease in a 43-year-old man is presented. He was asymptomatic, but the disease was recognized due to a routine chest roentgenographic study. Anemia, multiple lymphadenopathy, hypoalbuminemia and polyclonal hypergamma-globulinemia were observed. Histological examination of cervical lymph nodes revealed the plasma cell type of Castleman's disease. The diagnosis of PHG was confirmed by video-assisted thoracoscopical lung biopsy, and the immuno-histochemical staining of lamellar fibrosis for types I and III collagen was positive.

Adult↗
PubMed1994

[Plasma N-methylhistamine levels following intravenous administration of pipecuronium in man].

The half life of released plasma histamine (HA) is short but N-methylhistamine (N-MHA) which is a metabolite of HA has a longer life. Measuring both HA and N-MHA, it is possible to know the release of HA in the plasma. The purpose of this study was to ascertain the release of HA by pipecuronium (PPB) by measuring both HA and N-MHA in the plasma. Ten patients received 0.04 mg.kg-1 and other 10 patients received 0.08 mg.kg-1 of intravenous PPB. Plasma levels of HA and N-MHA were measured 1 minute before and 1, 3, 5 and 10 minutes after the administration of PPB. Release of plasma HA and N-MHA was not observed following administration of PPB. We concluded that PPB does not cause the release of histamine because there is no increase of the metabolite, N-MHA, of HA, and PPB can be used safely for patients with allergy.

Adult↗
PubMed1994

[Detection of Mycobacterium tuberculosis in transbronchial biopsy specimens by polymerase chain reaction].

Polymerase chain reaction (PCR) for the rapid detection of Mycobacterium tuberculosis was used to test 30 clinical specimens (transbronchial biopsy specimens) from patients in whom tuberculosis was suspected on chest X-ray. In these patients, 15 cases were histologically diagnosed as tuberculosis. Among them, M. tuberculosis was detected in 8 specimens by PCR. Of 8 PCR positive specimens, 7 were also positive by smear and culture; the other was negative by smear and positive by culture. On the other hand, 15 cases were therapeutically diagnosed as the tuberculosis and these specimens showed negative results in neither PCR nor microbiological tests. We conclude that the PCR method is useful for rapid and direct detection of M. tuberculosis in transbronchial biopsy specimens.

Biopsy↗
PubMed1994

Characterization of alternatively spliced PIG-A transcripts in normal and paroxysmal nocturnal hemoglobinuria cells.

The genetic lesion in paroxysmal nocturnal hemoglobinuria (PNH) cells resides in a DNA element that 1) encodes a product required for assembly of GlcNAc-inositol phospholipid and 2) is commonly affected in different patients. In this study, three alternative mRNA transcripts (1600, 1200, and 950 bp) that derive from this genetic element in normal cells were characterized. The 1200-bp transcript was found to arise from splicing out of 374 bp of exonic sequence extending from positions 407-780. The 950-bp transcript was found to arise from removal of this and 284 bp of additional exonic sequence beginning further upstream at position 123. Analyses of transcripts expressed in Epstein-Barr virus (EBV)-transformed B lymphocytes prepared from two PNH patients showed that both failed to express normal 1600-bp transcripts. One expressed truncated transcripts of 1000 and 800 bp generated by an alternate splice which utilized a downstream signal in place of the normal intronic splice signal. The other expressed a 1600-bp transcript with multiple nucleotide changes but normal 1200- and 950-bp "spliced" transcripts.

Alternative Splicing↗
PubMed1993

A candidate mammalian glycoinositol phospholipid precursor containing three phosphoethanolamines.

The glycoinositol phospholipid (GPI) anchors of mammalian proteins contain linear ethanolamine (EthN)-P-6ManManManGlcN glycan sequences that bear additional EthN-P substituents and in some cases include a fourth Man and a GalNAc or sialic acid-GalGalNAc. Precursors of these anchoring structures are preassembled in the endoplasmic reticulum by sequential glycosylation of inositol phospholipid. In previous studies (Hirose, S., Prince, G. M, Sevlever, D., Ravi, L., Rosenberry, T. L., Ueda, E., and Medof, M. E. (1992) J. Biol. Chem. 267, 16968-16974), a series of putative intermediates of this assembly process were isolated from human HeLa cells and murine lymphomas, and several of the more polar products were found to contain a second EthN-P attached to the Man residue (Man 1) proximal to GlcN. In this study, the most polar HeLa cell GPI species was purified by normal phase HeLa cell GPI species was purified by normal phase Iatrobead high performance liquid chromatography, and its glycan was characterized. Dionex anion exchange chromatographic analyses of fragments produced by nitrous acid deamination, hydrofluoric acid dephosphorylation, and trifluoroacetic acid hydrolysis in conjunction with biosynthetic labeling studies indicated a structure containing a third EthN-P substituent linked to the 6-position of Man 2. The polar GPI product exhibited a ManManManGlcN core lacking additional Man or GalNAc. The implications of the identification of this triply phosphoethanolamine-substituted species to mammalian GPI anchor biosynthesis are discussed.

Carbohydrate Sequence↗
PubMed1993

Deficient biosynthesis of N-acetylglucosaminyl-phosphatidylinositol, the first intermediate of glycosyl phosphatidylinositol anchor biosynthesis, in cell lines established from patients with paroxysmal nocturnal hemoglobinuria.

Paroxysmal nocturnal hemoglobinuria (PNH) is a hemolytic disorder caused by a deficiency of biosynthesis of the glycosyl phosphatidylinositol (GPI) anchor, but the biochemical defect is not completely understood. In the present study, we have analyzed affected cell lines established recently from two Japanese patients with PNH. Two lines of evidence indicate that these cells do not synthesize N-acetylglucosaminyl-phosphatidylinositol, the first intermediate in the GPI anchor biosynthesis. First, somatic cell hybridization analysis using Thy-1-deficient murine thymoma cell lines with known biochemical defects as fusion partners showed that the PNH cell lines belong to complementation class A, which is known not to synthesize N-acetylglucosaminyl-phosphatidylinositol. Second, analysis of in vitro glycolipid biosynthesis demonstrated that cell lysates of these PNH cell lines in fact did not support biosynthesis of N-acetylglucosaminyl-phosphatidylinositol. Thus, we have characterized for the first time the exact biochemical defect leading to PNH.

Cells, Cultured↗
PubMed1993

The expression of hemolytically active human complement protein C9 in mammalian, insect, and yeast cells.

The cDNA sequence encoding mature human C9 protein and its signal peptide was cloned into three expression vectors for expression in COS-7 (mammalian), Spodoptera frugiperda IPLB-SF-21AE (insect), and Saccharomyces cerevisiae (yeast) cells. In addition, C9 cDNA encoding only the mature protein was fused to the yeast invertase leader sequence (SUC2) and cloned for expression in yeast. Under optimal conditions COS-7 and IPLB-SF-21AE cells secreted recombinant C9 (rC9) at concentrations of about 111 and 700 ng C9/ml culture supernatant, respectively. By comparison S. cerevisiae, whether transformed with C9 cDNA containing its native or yeast invertase leader sequence, secreted only very small amounts of rC9 (5-10 ng/ml). However, upon lysis concentrations of up to 500 ng/mg dry wt were found in yeast cells transformed with C9 cDNA. SDS-PAGE followed by Western blot analysis revealed COS-7 cell and S. cerevisiae expressed rC9 to have a MW similar to that of native C9 purified from human serum, while rC9 from IPLB-SF-21AE cells was about 4 kDa smaller. No hemolytic activity of S. cerevisiae secreted rC9 could be detected and the specific hemolytic activity of S. cerevisiae intracellular rC9 was also very low. However, the specific hemolytic activities of COS-7 and IPLB-SF-21AE secreted rC9 were indistinguishable from that of purified native human C9. Thus, for future studies on the structure and function of C9 where the production of large quantities of mutant protein would be desirable, the baculovirus-insect cell expression system appears to offer considerable advantages.

Animals↗
PubMed1993

Idiopathic pulmonary fibrosis: progression of honeycombing at thin-section CT.

PURPOSE: To evaluate the progression of honeycombing at computed tomography (CT) and to determine serial changes in the pattern of disease. MATERIALS AND METHODS: CT scans, obtained in 29 patients with idiopathic pulmonary fibrosis who had undergone at least two serial CT examinations 1-52 months apart, were reviewed. Twelve patients had received no treatment, and 17 patients had received corticosteroids. Initial and follow-up CT scans were evaluated independently and then directly compared with each other. RESULTS: Twenty-six of the 29 patients showed progression of honeycombing, which was variable at CT (median, 0.4% [range, 0%-11%] per month) but not significantly different between untreated and treated patients. Areas of ground-glass attenuation on CT scans preceded and were predictive of the development of honeycombing in that location. Corticosteroid treatment reduced areas of ground-glass attenuation, but honeycombing on CT scans was irreversible. CONCLUSION: Low-dose therapy with corticosteroids does not suppress alveolitis sufficiently to prevent continued deterioration of the alveolar structures.

Female↗
PubMed1993

Clinical evaluation of rapid serodiagnosis of pulmonary tuberculosis by ELISA with cord factor (trehalose-6,6'-dimycolate) as antigen purified from Mycobacterium tuberculosis.

Immunoglobulin G (IgG) antibodies against purified cord factor (trehalose-6,6'-dimycolate) prepared from Mycobacterium tuberculosis H37Rv were determined by enzyme-linked immunosorbent assay (ELISA), and its diagnostic usefulness was evaluated. Serum specimens from 65 patients with active pulmonary tuberculosis, 58 patients with inactive pulmonary tuberculosis, 36 patients with diseases other than tuberculosis, and 66 healthy adults were examined. Patients with active pulmonary tuberculosis showed significantly higher titers of IgG antibodies against cord factor than did other groups (p < 0.001). The antibody titer greater than 0.29 in absorption difference (492 to 630 nm) of 160-times diluted serum was set as positive in ELISA. For patients with active and untreated pulmonary tuberculosis, the ELISA had a sensitivity of 81% and a specificity of 96%. From these results, it was concluded that the detection of IgG antibodies against cord factor is useful for serodiagnosis of active pulmonary tuberculosis. It was also indicated that the anticord factor antibody titers decline to the normal level as a result of antituberculosis chemotherapy.

Adult↗
PubMed1993

[Effects of sevoflurane on hemodynamics during the induction of anesthesia compared with those of isoflurane, enflurane and halothane].

The effects of sevoflurane (S), isoflurane (I), enflurane (E) and halothane (H) on hemodynamics were studied in 50 patients during the inhalation of 1.5 MAC of each anesthetic before the surgery. Mean arterial pressure (MAP), heart rate (HR), cardiac index (CI), systemic vascular resistance index (SVRI) and stroke volume index (SVI) were measured noninvasively using the automatic blood pressure manometer and the ultrasonic Doppler method (Accucom). There were significant differences between E and H in MAP, CI, SVRI and SVI. The values of CI, SVRI, SVI with S, as well as with I, were between those of E and H. These results indicate that sevoflurane causes the depression of blood pressure mostly by decreasing afterload during the induction of anesthesia, although the decrement with sevoflurane, as well as that with isoflurane, is less than that with enflurane.

Adult↗
PubMed1993

[Clinical features of the patients with "primary infection" of Mycobacterium avium complex].

Clinical characteristics are analysed in patients with primary infection of Mycobacterium avium complex (MAC). The definition of primary infection of MAC are determined as follows; 1) MAC is found several times since the beginning of the disease, 2) clinical symptoms or abnormal shadows on chest roentgenogram corresponding to MAC infection, 3) no old tuberculous lesions nor other abnormal shadows like bronchectasis, 4) no abnormal serological results suggesting other bacterial or viral infections. According to this definition, 17 out of 84 MAC patients are diagnosed as primary MAC infection, and clinical features are analyzed in these 17 patients. Average age of patients is 61.1 +/- 12.9 year old. This age is significantly higher than that of inpatients with pulmonary tuberculosis in our hospital, and lower than that of all MAC patients including primary and secondary infection. Five (29.4%) are male and 12 (70.6%) are female, the ratio of male to female is 1 to 2.4. This value is significantly different with that of inpatients with pulmonary tuberculosis in our hospital which revealed about 3 to 1. Most of the patients complained of cough with sputum, especially of hemosputum. Eleven patients (64.7%) out of 17 patients complained of repeated hemosputum. The frequency of hemosputum is very high compared with that of the patients with pulmonary tuberculosis (about 20%). No compromised condition was present except for a patient with Behcet's disease who was taking steroid hormone. Roentgenographic features of primary infection of MAC are those of scattered small nodular lesions in the peripheral part of the lung, thin wall cavity formation, no contraction of the diseased lung nor dislocation of the trachea.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗