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Biomedical subjects

M Ueda

Publications and source records attributed to M Ueda.

At least 433 records · Page 24Linked to original sources
PubMed1998

Detection of anti-type VII collagen antibody in Sjögren's syndrome/lupus erythematosus overlap syndrome with transient bullous systemic lupus erythematosus.

Bullous systemic lupus erythematosus (SLE) is a chronic, widespread, non-scarring, subepidermal blistering eruption associated with autoimmunity to type VII collagen. We describe a patient with Sj ogren's syndrome/lupus erythematosus overlap syndrome who showed transient blistering eruptions over limited skin surface and in oral mucosa. At the time of aggravation, the patient's serum contained IgG autoantibodies that bound to the dermal side of 1 mol/L NaCl-split normal skin, as determined by an indirect immunofluorescence test, and that reacted to type VII collagen, as determined by immunoblotting on dermal extract. Our observations suggest that a chronic, widespread, blistering eruption is not a prerequisite for the diagnosis of bullous SLE, and a mild, transient, blistering eruption could be a manifestation of type I bullous SLE.

Adult↗
PubMed1998

Pathophysiological response of cytokines and vasoactive agents in patients undergoing total gastrectomy.

OBJECTIVE: To investigate the involvement of vasoactive agents, endothelin (ET)-1, and atrial natriuretic peptide (ANP), and the responses of cytokines in patients undergoing total gastrectomy. DESIGN: Prospective study. SETTING: University hospital, Japan. SUBJECTS: 20 patients with advanced gastric cancer who had undergone total gastrectomy with lymph node dissection. INTERVENTIONS: Serum or plasma samples collected on the day before the operation, at the time of skin closure, and on postoperative days 1, 3, 5, and 7. MAIN OUTCOME MEASURES: Concentrations of acute phase reactants, cytokines (interleukin (IL)-1, tumour necrosis factor (TNF) and interleukin (IL)-6), and vasoactive agents (ET-1 and ANP). RESULTS: There were significant increases in concentrations of IL-6 and acute phase reactants postoperatively. ET-1 and ANP concentrations did not change significantly. CONCLUSION: There was no correlation between concentrations of the vasoactive agents ET-1 and ANP, and those of acute phase reactants or cytokines in serum or plasma in patients undergoing total gastrectomy.

Acute-Phase Proteins↗
PubMed1998

A monoclonal antibody, HCL-2, raised against human luteal cells reacts with apolipoprotein-B and detects the uptake of low density lipoprotein by luteinizing granulosa cells.

A monoclonal antibody, HCL-2, was raised by immunizing mice against human luteal cells. HCL-2 reacted with luteal cells and villous trophoblasts. The sodium dodecyl sulphate-polyacrylamide gel electrophoresis profile of immunopurified antigens from corpus luteum, chorionic villi, and placenta showed the same main protein band, the molecular mass of which is >200 kDa. The sequence of a portion of the N-terminal region of the antigenic protein purified from placenta was identical to that of apolipoprotein-B. The antigen purified from human serum and low density lipoprotein (LDL) using HCL-2 showed the same protein band as that from corpus luteum. Furthermore, the amino acid sequence (20 amino acids) of the protein purified from serum was also identical to that of apolipoprotein-B. Thus, we concluded that HCL-2 antigen is apolipoprotein-B. Human luteinizing granulosa cells isolated from the patients undergoing in-vitro fertilization treatment were cultured in the medium containing lipoprotein-deficient serum with or without supplementation of LDL. Using HCL-2, apolipoprotein-B was immunocytochemically detected on granulosa cells only in the presence of LDL. These findings showed that the uptake of LDL by granulosa cells was detected by immunocytochemical staining of apolipoprotein-B, indicating that HCL-2 is useful for analysing dynamic utilization of LDL by ovarian cells.

Adult↗
PubMed1998

Human large luteal cells in the menstrual cycle and early pregnancy express leukotriene A4 hydrolase.

Leukotriene (LT) A4 hydrolase (EC3.3.2.6) converts LTA4 to LTB4 which shows a chemotactic activity to leukocytes. To investigate the involvement of LTB4 in human corpus luteum (CL) function, the localization of LTA4 hydrolase in human ovarian tissues was examined by immunohistochemistry with a rabbit polyclonal antibody against LTA4 hydrolase. The enzyme was weakly expressed on granulosa cells of the follicles. After ovulation, the intensity of LTA4 hydrolase on large luteal cells increased and was highest in the midluteal phase. High expression was also observed in the CL of early pregnancy. In theca interna and small luteal cells, LTA4 hydrolase was weakly detected in all developmental stages. Immunoblot analysis demonstrated that the molecular mass of LTA4 hydrolase expressed in CL was 62 kDa, and confirmed that LTA4 hydrolase expression increased during CL formation and remained high in early pregnancy. The sequence encoding mRNA of LTA4 hydrolase, which was isolated from CL and amplified by polymerase chain reaction, was shown to be identical to the previously reported one. Immunocytochemistry showed that LTA4 hydrolase expression in cultured granulosa cells increased over 4 days in vitro and was enhanced by human chorionic gonadotrophin treatment. These expression profiles of LTA4 hydrolase suggest the involvement of LTB4 in luteal cell function during CL formation and early pregnancy.

Adolescent↗
PubMed1998

Human pancreatic RNase1-human epidermal growth factor fusion: an entirely human 'immunotoxin analog' with cytotoxic properties against squamous cell carcinomas.

The gene encoding human pancreatic ribonuclease 1 (hpRNasel) was fused with a gene encoding human epidermal growth factor (hEGF). The hybrid human protein was isolated from Escherichia coli inclusion bodies, refolded and purified to homogeneity. The fusion protein competed with 125I-hEGF for binding to hEGF receptors (EGFR) and had ribonucleolytic activities approaching those of hpRNase1. Several conformations having different enzymatic activities could be detected after reversed-phase high-performance liquid chromatographic analysis, the less hydrophobic molecules being the most active. The hybrid protein was specifically cytotoxic to A431, an EGFR overexpressing squamous carcinoma cell line, with an IC50 of approximately 10(-7) M. In contrast, recombinant hpRNase1 had an IC50 higher than 10(-4) M. A mixture of free hEGF and free hpRNasel was not more cytotoxic than hpRNasel alone and no cytotoxicity was detected in EGFR-deficient control cells. Taken together, these data suggest that this construct might be useful for targeted therapy of esophageal, lung and other squamous cell carcinomas and also breast cancers overexpressing EGFR, which correlate with a poor prognosis and cannot be cured by surgery alone. Engineering hybrid molecules with endogenous human proteins for targeted therapy may alleviate the dose-limiting immunogenicity and toxicity of conventional immunotoxins.

Binding, Competitive↗
PubMed1998

Basal cell carcinoma cells resemble follicular matrix cells rather than follicular bulge cells: immunohistochemical and ultrastructural comparative studies.

To detail the histogenetic relationship between basal cell carcinoma (BCC) and hair follicles, we immunohistochemically compared BCC cells to follicular matrix cells and follicular bulge cells using a panel of monoclonal antibodies against melanocytes, cytokeratins, subepidermal extracellular matrix components, and bullous pemphigoid (BP) sera, as well as using electron microscopy. Cytokeratin expression patterns were not consistent with the variety in types of cytokeratins and in cases of BCC. The distribution of some extracellular matrix components was not only linear along the interfaces of BCC tumor nests and stroma, and follicular matrix and follicular papilla; granular deposits were also seen in the stroma and follicular papilla, whereas they were only linearly distributed along the follicular bulge. The BP antigens and integrin alpha 6, which were absent in BCC and follicular matrix, were expressed in the follicular bulge area. Electron microscopically, hemidesmosomes were poorly organized in these three tissues, but the lamina densa was incomplete in BCC and follicular matrix, whereas the lamina densa in the follicular bulge area was continuous. These morphologic similarities between BCC and follicular matrix cells, and coexistence of melanocytes in the BCC tumor nest strongly suggest the differentiation of BCC toward the follicular matrix cells.

Basal Cell Carcinoma↗
PubMed1998

Effects of sex steroids and growth factors on invasive activity and 5'-deoxy-5-fluorouridine sensitivity in ovarian adenocarcinoma OMC-3 cells.

Effects of sex steroids (estradiol-17 beta, E2; progesterone, Prog) and growth factors (epidermal growth factor, EGF; transforming growth factor-alpha, TGF-alpha) on invasive activity and 5'-deoxy-5-fluorouridine (5'-dFUrd) sensitivity of ovarian adenocarcinoma OMC-3 cells were investigated. Tumor cell migration along a gradient of substratum-bound fibronectin and invasion into reconstituted basement membrane were inhibited by 10 microM Prog, but stimulated by 0.1-10 nM EGF and TGF-alpha in a concentration-dependent manner. E2 did not have any effect on tumor cell migration or invasion. The zymography of tumor conditioned medium showed that the treatment of OMC-3 cells with EGF and TGF-alpha resulted in increases of type IV collagenase, stromelysin and urokinase-type plasminogen activator (uPA). EGF and TGF-alpha up-regulated thymidine phosphorylase (dThdPase) expression of tumor cells and consequently enhanced the antiproliferative action of 5'-dFUrd, which is converted to 5-fluorouracil by dThdPase. E2 and Prog did not have significant effects on the expression of proteolytic enzymes and dThdPase, or on the 5'-dFUrd sensitivity of tumor cells. The inhibitory effect of Prog on tumor cell invasion may depend on its inhibitory action on the motility of tumor cells. These results suggest that EGF and TGF-alpha simultaneously up-regulate the potential of ovarian adenocarcinoma cells to invade extracellular matrices and their dThdPase expression, both of which are associated with the specific action of 5'-dFUrd selectively to kill tumor cells with high invasive and metastatic potential.

Basement Membrane↗
PubMed1998

Effect of gelonin immunoconjugate with monoclonal antibody MSN-1 to endometrial adenocarcinoma on antigen-producing tumor cells in vivo.

Missile therapy, which destroys cancer cells specifically, has been advocated as an effective modality for the treatment of carcinoma. We have developed an immunoconjugate consisting of the monoclonal antibody MSN-1 (IgM), which reacts strongly with endometrial adenocarcinomas, combined with a plant hemitoxin named gelonin via a disulfide bond using N-succinimidyl-3-(2-pyridyldithio) propionate and 2-iminothiolane, and examined its selective cytotoxicity in athymic mice. The reductions in resected weights of target tumor cells, at the local site of MSN-1-gelonin immunoconjugate treatment, were 96% with local administration and 75% with caudal vein administration, as compared with the untreated group. There was no weight loss in treated mice. Our results suggest that this MSN-1-gelonin immunoconjugate has potential clinical applications in the treatment of endometrial adenocarcinomas.

Adenocarcinoma↗
PubMed1998

A study of sports-related mandibular angle fracture: relation to the position of the third molars.

Mandibular angle fractures have been considered attributable to the presence of unerupted third molars. We examined the relationship between the incidence of sports-related mandibular angle fractures and the presence of a mandibular third molar with emphasis on the position of the third molar. The incidence of angle fracture was significantly higher in the sports-related injury group than in the group with fracture due to other causes (P < 0.05). The incidence of angle fracture in the athletes with higher impaction scores was significantly higher than that in the subjects with higher scores who did not have sports-related fractures (P < 0.05). The percentage of rugby athletes with third molars was significantly higher than that of those without third molars (P < 0.001), and a high proportion of young athletes (89.5%) was considered to belong to a potential high-risk group for angle fractures. Our findings suggest that mandibular angle fractures are influenced by the presence and characteristics of the third molar in sports-related injuries.

Accidents, Traffic↗
PubMed1998

Oral injury and mouthguard usage by athletes in Japan.

The use of mouthguards in contact sports effectively prevents oral injury and preserves oral structures. We investigated oral injuries and awareness concerning mouthguards among Japanese high school soccer and rugby players. Athletes were asked a series of questions concerning their history of oral injury while participating in sports, and their pattern of mouthguard use was determined. The data were evaluated statistically using chi-square analysis. The incidence of oral injuries was 32.3% for soccer athletes and 56.5% for rugby athletes, with 0.8% and 24.1% of the respective groups having mouthguards. There were significant differences between the soccer and rugby groups (P < 0.001). Although 81.8% of soccer athletes thought mouthguards were unnecessary, only 26.2% of rugby athletes shared this opinion and there was a significant difference between the soccer and rugby groups. Many soccer athletes had insufficient knowledge about mouthguards and were not concerned about preventing oral injury, although it was in fact a common problem in their sport. Athletes as well as coaches must be made aware of the high risk of oral injury when playing soccer, rugby, and other contact sports.

Adolescent↗
PubMed1998

A new intermaxillary fixation method using adhesive cast splints for avoiding skin puncture.

Intermaxillary fixation using arch bars is performed for mandibular and maxillary fractures. They are applied to the respective maxillary and mandibular arches with circumdental stainless steel wires. Skin puncture may pose a risk for the surgeon. Therefore, the authors have developed and tested a method for using intermaxillary fixation using an adhesive cast splint. The resulting strength was found to be generally superior to that achieved with arch bars. Despite some problems, this technique is considered a useful method for avoiding skin puncture.

Adhesives↗
PubMed1998

[Evaluation of pyuria by means of urinary sediment method--a comparison to the counting chamber method].

We compared the urinary sediment method to counting chamber method in evaluating pyuria, in order to elucidate an issue of urinary sediment method. The KOVA system was used for counting chamber method. Three hundred and ten urine specimens from the patients who visited Fuji City Central Hospital in March, 1995, were employed. The numbers of leukocytes of each specimen was counted by means of urinary sediment method and counting chamber method, and compared. The urinary leukocyte count evaluating by means of these two methods were correlated well. However, the discrepancy of urinary leukocyte counts by means of these two methods were recognized in two specimens. In these two specimens, the leukocyte count was 1 to 4/5HPF by means of urinary sediment method, but more than 50/microliter by means of counting chamber methods. Significant bacteria (10(4) cfu/ml or more), positive esterse reaction, and high pH (8.5) were recognized in both specimens. In addition, one of two these specimens was hypotonic (specific gravity was 1.008). Therefore, it was suggested that leukocytes were sometimes disrupted with centrifugation. These results indicate that counting chamber method is more useful urinary sediment method to detect pyuria.

Adolescent↗
PubMed1998

[Correlation in Staphylococcus aureus infections between toxic shock syndrome toxin-1 production and clinical feature].

Toxic shock syndrome (TSS) is caussed by toxic shock syndrome toxin-1 (TSST-1) of Staphylococcus aureus. We studied the incidence of TSST-1 production by the clinical isolates of S. aureus, in order to clarify the possibility of TSS caused by S. aureus. One hundred and seventeen clinical isolates of S. aureus were tested. Of 117 strains, 74 were methicillin-cephem resistant S. aureus (MRSA), and 43 were methicillin-cephem sensitive S. aureus (MSSA). TSST-1 production and penicillinase (PCase) activity were measured by means of reversed passive Latex aggulutination method and acidmetric assay, respectively. The incidences of TSST-1 production by PCase positive MRSA, PCase negative MRSA, PCase positive MSSA and PCase negative MSSA were 92% (55/60), 78% (11/14), 21% (7/33) and 0% (0/10), respectively. TSS occurred in a patient with chronic complicated pyelonephritis after endopyelotomy for hydronephrosis. In this patient, PCase positive MRSA which produced TSST-1 was isolated from urinary tract, and he was cured after administration of arbekacin. These results indicsate that methicillin-cephem resistance and PCase production are the risk factors for TSST-1 production in S. aureus.

Bacterial Toxins↗
PubMed1998

Assimilation of cellooligosaccharides by a cell surface-engineered yeast expressing beta-glucosidase and carboxymethylcellulase from aspergillus aculeatus

Since Saccharomyces cerevisiae lacks the cellulase complexes that hydrolyze cellulosic materials, which are abundant in the world, two types of hydrolytic enzymes involved in the degradation of cellulosic materials to glucose were genetically co-immobilized on its cell surface for direct utilization of cellulosic materials, one of the final goals of our studies. The genes encoding FI-carboxymethylcellulase (CMCase) and beta-glucosidase from the fungus Aspergillus aculeatus were individually fused with the gene encoding the C-terminal half (320 amino acid residues from the C terminus) of yeast alpha-agglutinin and introduced into S. cerevisiae. The delivery of CMCase and beta-glucosidase to the cell surface was carried out by the secretion signal sequence of the native signal sequence of CMCase and by the secretion signal sequence of glucoamylase from Rhizopus oryzae for beta-glucosidase, respectively. The genes were expressed by the glyceraldehyde-3-phosphate dehydrogenase promoter from S. cerevisiae. The CMCase and beta-glucosidase activities were detected in the cell pellet fraction, not in the culture supernatant. The display of CMCase and beta-glucosidase proteins on the cell surface was confirmed by immunofluorescence microscopy. The cells displaying these cellulases could grow on cellobiose or water-soluble cellooligosaccharides as the sole carbon source. The degradation and assimilation of cellooligosaccharides were confirmed by thin-layer chromatography. This result showed that the cell surface-engineered yeast with these enzymes can be endowed with the ability to assimilate cellooligosaccharides. This is the first step in the assimilation of cellulosic materials by S. cerevisiae expressing heterologous cellulase genes.

Journal Article↗
PubMed1998

Cytotoxic T-lymphocyte-mediated lysis of Toxoplasma gondii-infected target cells does not lead to death of intracellular parasites.

CD8(+) T cells play a crucial role in the control of infection with intracellular microbes. The mechanisms underlying the CD8(+) T-cell-mediated clearance of the intracellular pathogen Toxoplasma gondii are, however, not completely understood. The effect of CD8(+) cytotoxic T-lymphocyte (CTL)-mediated lysis of host cells on the viability of intracellular T. gondii was investigated. Quantitative competitive PCR of the gene encoding T. gondii major surface antigen (SAG-1) was combined with treatment of the parasites with DNase, which removed the DNA template of nonviable parasites. The induction by CD8(+) CTLs of apoptosis in cells infected with T. gondii did not result in the reduction of live parasites, indicating that intracellular T. gondii remains alive after lysis of host cells by CTLs.

Animals↗
PubMed1998

Genetic evaluation of physiological functions of thiolase isoenzymes in the n-alkalane-assimilating yeast Candida tropicalis.

The n-alkane-assimilating diploid yeast Candida tropicalis possesses three thiolase isozymes encoded by two pairs of alleles: cytosolic and peroxisomal acetoacetyl-coenzyme A (CoA) thiolases, encoded by CT-T1A and CT-T1B, and peroxisomal 3-ketoacyl-CoA thiolase, encoded by CT-T3A and CT-T3B. The physiological functions of these thiolases have been examined by gene disruption. The homozygous ct-t1a delta/t1bdelta null mutation abolished the activity of acetoacetyl-CoA thiolase and resulted in mevalonate auxotrophy. The homozygous ct-t3a delta/t3b delta null mutation abolished the activity of 3-ketoacyl-CoA thiolase and resulted in growth deficiency on n-alkanes (C10 to C13). All thiolase activities in this yeast disappeared with the ct-t1a delta/t1bdelta and ct-t3a delta/t3bdelta null mutations. To further clarify the function of peroxisomal acetoacetyl-CoA thiolases, the site-directed mutation leading acetoacetyl-CoA thiolase without a putative C-terminal peroxisomal targeting signal was introduced on the CT-T1A locus in the ct-t1bdelta null mutant. The truncated acetoacetyl-CoA thiolase was solely present in cytoplasm, and the absence of acetoacetyl-CoA thiolase in peroxisomes had no effect on growth on all carbon sources employed. Growth on butyrate was not affected by a lack of peroxisomal acetoacetyl-CoA thiolase, while a retardation of growth by a lack of peroxisomal 3-ketoacyl-CoA thiolase was observed. A defect of both peroxisomal isozymes completely inhibited growth on butyrate. These results demonstrated that cytosolic acetoacetyl-CoA thiolase was indispensable for the mevalonate pathway and that both peroxisomal acetoacetyl-CoA thiolase and 3-ketoacyl-CoA thiolase could participate in peroxisomal beta-oxidation. In addition to its essential contribution to the beta-oxidation of longer-chain fatty acids, 3-ketoacyl-CoA thiolase contributed greatly even to the beta-oxidation of a C4 substrate butyrate.

Acetyl-CoA C-Acetyltransferase↗
PubMed1998

Application of subtype-specific monoclonal antibodies for rapid detection and identification of influenza A and B viruses.

We established a rapid method for the identification of influenza A and B virus strains: the peroxidase-antiperoxidase (PAP) staining method with two subtype-specific murine monoclonal antibodies, C179 (H1 and H2 specific) and F49 (H3 specific), and an anti-influenza B virus rabbit polyclonal serum. The types and subtypes of 160 strains were examined, and 158 strains were identified to be the same by the hemagglutination-inhibition (HI) test and the PAP method. In contrast to the results by the HI test, two strains were revealed to be a mixture of two subtypes (H1 and H3) by the PAP method, which was confirmed by plaque cloning. We further analyzed clinical specimens by the PAP method by directly inoculating specimens into Madin-Darby canine kidney cells in microplates. After 40 h of incubation, the types and subtypes of viruses in 52 of 152 specimens were clearly identified. Since the reactivities of the two monoclonal antibodies are not influenced by the antigenic drift of influenza virus, the newly developed method should be applicable not only for rapid diagnosis but also for the epidemiological study of influenza.

Animals↗