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Biomedical subjects

M Ueda

Publications and source records attributed to M Ueda.

At least 73 records · Page 4Linked to original sources
PubMed1995

Establishment of a CD4+ T cell clone recognizing autologous hematopoietic progenitor cells from a patient with immune-mediated aplastic anemia.

In some patients with aplastic anemia (AA), hematopoietic function is dependent on continuous administration of cyclosporine A (CyA). These AA patients may have T lymphocytes whose myelosuppressive effect is mitigated by CyA. We established a total of 29 T cell clones from the bone marrow of a CyA-dependent AA patient in relapse. Some of the CD4+ T cell clones demonstrated a specific proliferative response to irradiated autologous bone marrow cells enriched for CD34+ cells (CD34(+)-rich cells) obtained from the patient in remission. One of the T cell clones showing the best proliferative response to CD34(+)-rich cells carried the T cell receptor V beta 17 and produced interferon-gamma (IFN-gamma) only when cultured with autologous CD34(+)-rich cells. This T cell clone inhibited colony formation by colony-forming unit-granulocyte/macrophage (CFU-GM) and burst-forming unit-erythroid (BFU-E) by approximately 60% when it was cultured with autologous CD34(+)-rich cells in methylcellulose medium, although the clone did not exhibit direct cytotoxicity to the CD34(+)-rich cells. The inhibition of in vitro hematopoietic progenitor cell growth by the T cell clone was partially abrogated by the addition of CyA to the culture. These findings suggest that in some patients with CyA-dependent AA, CD4+ T cells autoreactive to hematopoietic progenitor cells exist and may play an important role in the pathogenesis of bone marrow failure.

Adult↗
PubMed1994

Peptide synthesis with halophenylalanines by thermolysin.

Thermolysin was able to catalyze enantioselective peptide synthesis with non-natural amino acids, halophenylalanines. However, the reactivity of thermolysin was considerably influenced by the kind and position of halogen substituents on these analogues. The manner of the recognition of the amino component by the enzyme was different from that of the carboxyl component in the synthesis of peptides with non-natural phenylalanine analogues. The phenomena observed are discussed, based on the kinetic parameters obtained.

Amino Acid Sequence↗
PubMed1994

Characterization of the catalase of the n-alkane-utilizing yeast Candida tropicalis functionally expressed in Saccharomyces cerevisiae.

Candida tropicalis catalase (CTC) genomic DNA was recombined on a plasmid with the galactose-inducible GAL7 promoter and expressed highly as a heme protein in Saccharomyces cerevisiae as host. The percentage of recombinant CTC (rCTC) in total extractable protein amounted to at least 25%. The rCTC was purified and characterized in terms of subunit mass, behavior in native polyacrylamide gel electrophoresis, absorption spectrum, amino-terminal amino acid sequence, peptide map, specific activity, and Michaelis constant (Km) value for hydrogen peroxide. These properties were similar or identical to those of the purified enzyme from C. tropicalis (CTC). From these results, this system appears suitable for high expression of functional catalase protein having heme.

Base Sequence↗
PubMed1994

Ultrastructural and immunohistochemical characterization of basal cells in three-dimensional culture models of the skin.

Keratinocytes were cultured on fibroblast-free dermal substitutes made of type I collagen film (collagen dermal substitute) and an extracellular matrix gel film (matrix dermal substitute), each of which was laid on a lyophilized type I collagen sponge. The morphology of the basal keratinocytes in these three-dimensional culture models of the skin was studied ultrastructurally and immunohistochemically to assess their differentiation to basal cells. The basal keratinocytes in the artificial epidermis cultured on the collagen dermal substitute showed poorly organized tonofibril networks and desmosomes. Neither the tonofibril-hemidesmosome complex nor the lamina densa were detected along the interface, where many cytoplasmic projections of basal keratinocytes were noted. There were no detectable antigens of type IV or VII collagen, LDA-1, or laminin in the interface. Bullous pemphigoid (BP) and 1-2B7B antigens and integrins were expressed along the cytoplasmic membrane and the projections of the basal keratinocytes. A high molecular weight keratin (keratin 1, 68 kDa, 34 beta B4) was detected only in part of the uppermost layers of this artificial epidermis. In contrast, basal keratinocytes in the artificial epidermis on the matrix dermal substitute developed tonofibril networks radiating to desmosomes and hemidesmosomes, under which a primitive lamina densa was present. Basement membrane zone antigens, such as type IV and VII collagens, LDA-1 and laminin were noted along the interface as were 1-2B7B and BP antigens and integrins. Laminin and type VII collagen were also detected along or in the membrane of the endoplasmic reticulum of basal keratinocytes.(ABSTRACT TRUNCATED AT 250 WORDS)

Basement Membrane↗
PubMed1994

M-VEC (methotrexate, vinblastine, epirubicin, and cisplatin) with granulocyte colony-stimulating factor for the treatment of urothelial cancer: an effective and safe chemotherapy regimen.

The toxicity of combination chemotherapy is significant, with the most prominent side effect being myelosuppression. To reduce the toxicity, we used a recombinant human granulocyte colony-stimulating factor (rhG-CSF). A total of 52 patients were enrolled in this study. The sites of tumor involvement included the urinary bladder in 24 patients, the renal pelvis in 5, the ureter in 4, lymph nodes in 11, bone in 4, the lung in 1, and miscellaneous sites in 4 patients. The chemotherapy was given in 21-day cycles as follows: 30 mg/m2 methotrexate was given intravenously on day 1, and approximately 24 h later, 3 mg/m2 vinblastine, 30 mg/m2 epirubicin, and 70 mg/m2 cisplatin were given intravenously. The rhG-CSF (2 micrograms/kg per day) was injected subcutaneously on days 3-16 of each cycle. All patients received full doses of the antineoplastic agents on time according to the protocol design. The response rates were 61% for primary sites, 55% for lymph nodes, 0 for bone, and 67% for miscellaneous sites. Of 42 patients evaluated, 5 (12%) achieved a complete response and 20 (48%) achieved a partial response, for an overall response rate of 60%. Of the 42 patients, 27 (64%) are alive, and the median duration of survival is 14 months. The mean nadir white blood count was more than 5,600 cells/mm3. The incidence of mucositis in the total toxic symptoms was low. There was no cardiac toxicity or drug-related death. These results indicate that the present combination chemotherapy with coadministration of rhG-CSF is an effective and safe regimen for the treatment of urothelial cancer.

Adenocarcinoma↗
PubMed1994

Age and sex-dependent alterations of serum amylase and isoamylase levels in normal human adults.

Total amylase and salivary- and pancreatic-type isoamylase levels were assayed in sera from 606 apparently healthy adults of different sex and age groups. There were significant differences in both total amylase and isoamylase levels, depending on age and sex, one of the characteristics being that levels of these three enzymes were significantly higher in the elderly group in both men and women than in other age groups. Another feature was that all of these enzyme levels were significantly greater in women in the third and fourth decade than in men. Age and sex differences should be taken into consideration in the evaluation of mild hyperamylasemia.

Adolescent↗
PubMed1994

Evidence that glutamate is released from capsaicin-sensitive primary afferent fibers in rats: study with on-line continuous monitoring of glutamate.

The aim of this study is to elucidate whether the excitatory amino acid glutamate is released from capsaicin-sensitive primary afferent fibers, and to compare the releasing effect of capsaicin on glutamate with that on substance P. The release of glutamate was measured using a fluorometric on-line continuous monitoring system, in which the immobilized glutamate dehydrogenase column was connected to an in vitro superfusion system. In the presence of 0.3 microM tetrodotoxin, 2-min application of capsaicin produced an increased outflow of glutamate, as well as an increase in the release of immunoreactive substance P from dorsal horn slices of the rat. The release of glutamate was concentration-dependently increased by capsaicin at concentrations in the range of 0.1-3 microM, and the release evoked by 10 microM capsaicin was not higher than that evoked by 3 microM. On the other hand, capsaicin at concentrations of 1-10 microM produced a concentration-dependent increase in the release of immunoreactive substance P, without effect at 0.1 microM. The amount of glutamate release evoked by 3 microM capsaicin was about 42.8 pmol.mg-1 protein, and 290 times that of immunoreactive substance P. The release of glutamate by 3 microM capsaicin was suppressed by the depletion of calcium from the superfusate. Capsaicin at 3 microM failed to increase the release of glutamate from the dorsal horn slices of the rats made an L4-L6 dorsal rhizotomy. These results suggest that capsaicin evoked the release of glutamate from primary afferent fibers in the dorsal horn and that glutamate may play an important role in pain transmission between primary afferent fibers and dorsal horn neurons.

Animals↗
PubMed1994

SENCAR mouse skin tumors produced by promotion alone have A to G mutations in codon 61 of the c-rasHa gene.

SENCAR mice, developed by selective breeding for high susceptibility to skin carcinogenesis by initiation with 7,12-dimethylbenz[a]anthracene and promotion with 12-O-tetradecanoylphorbol-13-acetate (TPA), form squamous papillomas in approximately 20% of animals treated repeatedly with TPA, without chemical initiation. DNA from eight skin tumors produced by a TPA-only protocol and four cell lines derived from these tumors was amplified by polymerase chain reaction and analyzed by discriminative oligonucleotide hybridization using oligomers specific for various c-rasHa gene codon 61 sequences. Five tumors and three cell lines had CAA (wild-type) to CGA mutations. In addition, one tumor had a CAA to CTA mutation, for a total of six of eight tumors having an activating mutation at this codon. Two tumors and one cell line had no codon 61 mutations detectable by this method. Since tumors derived from promotion-only protocols presumably originated from constitutively initiated cells, we examined tumor-free skins of untreated newborn and eight-month-old retired breeders and of 78-88-week-old SENCAR mice of both sexes, which were treated with TPA for 10 weeks starting at age 16-28 weeks and were untreated thereafter. Only the wild-type c-rasHa gene codon 61 sequence was seen, suggesting that the constitutively initiated cell population, if present, is below the limit of detection by this method.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1994

Lack of low Km diazepam N-demethylase in livers of poor metabolizers for S-mephenytoin 4'-hydroxylation.

Metabolism of diazepam was studied in vitro to identify the forms of cytochrome P450 (CYP) responsible for N-demethylation (nordazepam formation) and 3-hydroxylation (temazepam formation), using liver microsomes obtained from extensive (EM) and poor metabolizers (PM) for S-mephenytoin 4'-hydroxylation. Involvement of at least two P450 forms in diazepam N-demethylation was suggested by a biphasic pattern in Lineweaver-Burk and Eadie-Hofstee plots from the EM, whereas a monophasic pattern was observed from the PM liver microsomes. The kinetic parameters for the N-demethylation in the EM group were: Km 1, 19.4 +/- 0.4 microM; Vmax 1, 0.27 +/- 0.04 nmol min-1 per mg protein; Km 2, 346 +/- 34 microM; Vmax2, 1.82 +/- 0.63 nmol min-1 per mg protein (n = 3, mean +/- SD). The PM group showed the mean values of Km and Vmax (Km, 319 +/- 30 microM; Vmax, 1.49 +/- 0.62 nmol min-1 per mg protein) (n = 3) similar to those of Km2 and Vmax2 in the EM group. An antibody raised against CYP2C9 (anti-human CYP2C) strongly inhibited diazepam N-demethylation in EM liver microsomes at a low substrate concentration (20 microM). However, the anti-human CYP2C showed no clear inhibition of N-demethylation in EM liver microsomes at a high substrate concentration (200 microM). Diazepam N-demethylation in PM liver microsomes was not clearly inhibited by the anti-human CYP2C at either the low or high substrate concentrations. These data suggest that different P450 forms mediated diazepam N-demethylation in EM and PM liver microsomes, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies↗
PubMed1994

A case of testicular teratoma located in the opposite side of the upper abdominal wall.

We report a case of a testicular teratoma located in the opposite side of the upper abdominal wall. A 2-year-old boy presented with a non-palpable right testis and underwent right inguinal exploration for right cryptorchidism. During surgery, the tumor was found in the opposite side of the upper abdominal wall across the midline. Histological investigation of the tumor revealed a mature teratoma occurring in the intra-abdominal testis. No such case has been previously reported.

Child, Preschool↗
PubMed1994

M-VEC (methotrexate, vinblastine, 4'-epirubicin and cisplatin) combined with glycosylated recombinant human granulocyte colony-stimulating factor (rhG-CSF) for the treatment of transitional cell carcinoma of urothelium: reduction in toxicity produced by rhG-CSF.

Forty-six patients with urothelial cancer were treated with a systemic chemotherapeutic regimen consisting of methotrexate, vinblastine, 4'-epirubicin and cisplatin (M-VEC) in conjunction with glycosylated recombinant human granulocyte colony stimulating factor (rhG-CSF); then 33 were evaluated for response. Complete response was observed in 7 patients (21%) and partial response in 13 (39%). As far as the toxic effects of this treatment are concerned, mucositis of a minimum grade and leukopenia greater than grade 3 occurred in 5% and 10% of the patients, respectively; there were no cases of nadir sepsis and drug-related death. Minor toxicity such as nausea vomiting occurred in 81% of patients, and no patient required either dose-reduction or a delay of more than 5 d before starting of the second cycle. Thus, it may be concluded that M-VEC chemotherapy combined with rhG-CSF is useful in the treatment of urothelial cancer, especially when used as a neoadjuvant.

Adult↗
PubMed1994

Macrophage activation factors and cytokines in peritoneal fluid from patients with endometriosis.

Endometriosis is often associated with infertility, which might be a consequence of intraperitoneal macrophage activation and various immunological events related to it. To determine what triggers macrophage activation and what immunologic conditions occur in the peritoneal cavity of a patient with endometriosis, we measured the amounts of phospholipase A2, cholesterol fractions (HDL, LDL, and VLDL), interleukins (IL)-1 and 6, granulocyte-macrophage-colony stimulating factor (GM-CSF), and phosphatidylserine in peritoneal fluid. Peritoneal-fluid samples were collected during laparoscopy or laparotomy from 14 patients with endometriosis and from 11 patients with other diseases (5 with myoma uteri, 5 with benign ovarian cysts and 1 with a double uterus) as controls. With regard to phospholipase A2, there were no significant differences between the endometriosis group and the control group. LDL was significantly lower in the endometriosis group than in the control (p < 0.05). The levels of IL-1 and GM-CSF were similar in both groups; IL-6, however, was higher in 7 endometriosis patients with associated infertility than in the 9 non-endometriosis fertile patients. These findings suggest that inflammatory tissues are not related to the activation of intraperitoneal macrophages in endometriosis patients. Cholesterol fractions (HDL or LDL) might participate in macrophage activation in these patients, but this remains to be clarified. The high IL-6 levels observed in infertile endometriosis suggest that some immunological events that prevent patients from becoming pregnant take place in women with endometriosis-associated infertility.

Adult↗
PubMed1994

Evidence that Escherichia coli ubiA product is a functional homolog of yeast COQ2, and the regulation of ubiA gene expression.

The Escherichia coli ubiA gene coding for 4-hydroxy benzoate octaprenyl transferase is thought to be a key enzyme of ubiquinone biosynthesis. Strains with ubiA disrupted were constructed by chromosomal gene replacement with the chloramphenicol resistance gene. The respiration-defective phenotype of the ubiA mutant was complemented by expression of the COQ2 gene encoding the 4-hydroxy benzoate hexaprenyl transferase of Saccharomyces cerevisiae and such strains produced ubiquinone-8. This strongly supports the idea that COQ2 catalyzes the same enzymatic reaction with UbiA and the substrate specificity of COQ2 is broad. Study of the expression of ubiA using an ubiA-lacZ fusion system showed that the ubiA expression was catabolite-repressed by glucose. This repression by glucose was obvious in the arcA mutant. ArcA is the positively acting transcriptional regulator of the oxygen regulated genes. The molecular mass of the protein product of ubiA was 32kD, found using the over-expression of the ubiA gene.

Base Sequence↗
PubMed1994

Purification of inulin fructotransferase (DFA I-producing) from Arthrobacter sp. MCI2493 and production of DFA I from inulin by the enzyme.

An extracellular enzyme that produces di-D-fructofuranose-2',1;2,1' dianhydride from inulin was purified from the culture broth of Arthrobacter sp. MCI2493. The molecular weight of the enzyme was 40,000 by gel filtration and SDS polyacrylamide gel electrophoresis. The enzyme had maximum activity at pH 6.0 and 50 degrees C. Using this purified enzyme, 100 g/liter inulin was converted into 60 g/liter of DFA I, nystose, and 1-F-fructofuranosyl-nystose after incubation for 30 h.

Arthrobacter↗
PubMed1994

Isolation of naturally processed peptides from a Toxoplasma gondii-infected human B lymphoma cell line that are recognized by cytotoxic T lymphocytes.

Naturally processed peptides derived from Toxoplasma gondii (T. gondii) were acid extracted from T. gondii-infected cells and detected by cytotoxic T lymphocytes (CTL) derived from peripheral blood lymphocytes of a patient with chronic toxoplasmosis. The CTL lines were obtained by weekly in vitro stimulation with a T. gondii-infected human B cell lymphoma line, ARH, which shares HLA-A2 and -Cw4 determinants with the patient. The lytic activity of these CTL lines against T. gondii-infected ARH and ARH pulsed with fraction 29 of a reversed-phase high-performance liquid chromatography (HPLC) extract from T. gondii-infected ARH was inhibited by an anti-HLA-A, B, C monoclonal antibody (mAb) and an anti-HLA-A2 mAb. Anti-HLA-DR mAb failed to block the lytic activity. Thus, the presentation of peptides by T. gondii-infected cells for CTL is mediated by HLA-A2 molecules. Interestingly, antigen presentation of ARH pulsed with naturally processed HPLC fraction 29 peptides was not inhibited by treatment with brefeldin A. The amino acid sequence of the HLA-A2-bound peptide in fraction 29 was in part consistent with the predictive algorithm of HLA-A2-binding peptide motifs.

Amino Acid Sequence↗
PubMed1994

Clinical evaluation of the Endosearch sampler in endometrial cytology-a preliminary report.

The clinical value of the Endosearch endometrial cell sampling device was assessed in 94 cases managed in our outpatient clinic. The Endosearch could be easily inserted into the uterine cavity in 91 (96.8%) of 94 cases without analgesia. Pain and prolonged bleeding accompanied by the insertion of the Endosearch occurred in eight (8.8%) and six (6.6%) of 91 cases, respectively. Satisfactory material for cytologic diagnosis of the endometrial state was obtained in 90(98.9%) of 91 cases, which were comparable to those with the Endocyte. On the other hand, the endometrial tissue samples could be simultaneously obtained with the Endosearch in 78 (85.7%) of 91 cases, and histologic diagnosis was possible in 69(88.5%) of 78 cases. Among them, three of five cases with endometrial hyperplasia and four of five cases with endometrial carcinoma could be correctly diagnosed by the Endosearch histologically. These results suggest that outpatient investigation of the endometrial state by the Endosearch sampler is quite useful not only in cytologic diagnosis but also in histologic evaluation of endometrial lesions.

Aged↗