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Biomedical subjects

M Ueda

Publications and source records attributed to M Ueda.

At least 91 records · Page 5Linked to original sources

Intercellular adhesion molecule-1 and leukocyte function-associated antigen-1 are involved in protection mediated by CD3+TCR alpha beta- T cells at the early stage after infection with Listeria monocytogenes in rats.

To investigate the significance of intercellular adhesion molecule-1 (ICAM-1) and leukocyte function-associated antigen-1 (LFA-1) in host defense against infection with intracellular parasites, we examined the effects of in vivo pretreatment with mAbs to ICAM-1 (1A29) and LFA-1 alpha (WT-1) on the protection against infection with Listeria monocytogenes in Fisher F344/N rats. Expression of ICAM-1 and LFA-1 alpha molecules on T cells in spleen, liver and peritoneal cavity of rats was down-regulated after i.p. administration with daily doses of 300 micrograms of either 1A29 or WT-1 for 10 days. The survival rate of rats inoculated with viable Listeria was significantly reduced by in vivo pretreatment with 1A29 together with WT-1 for 10 days but not by in vivo pretreatment with control mAb. The numbers of bacteria in the spleen in rats pretreated with both 1A29 and WT-1 were significantly increased on day 3 and day 6 after infection with 1 x 10(7) of viable Listeria corresponding to 1/30 of LD50 to normal rats. Thus, the resistance against listerial infection was severely impaired by combinational pretreatment with mAbs in ICAM-1 and LFA-1 alpha. As shown in our previous report, the early appearance of CD3+TCR alpha beta- T cells, presumably TCR gamma delta T cells, was evident in the peritoneal cavity and liver of control rats at the early stage after listerial infection, while this was suppressed at this stage in rats pretreated with both 1A29 and WT1.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

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

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

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

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

Cytokines stimulate dipeptidyl peptidase-IV expression on human luteinizing granulosa cells.

We have previously reported that dipeptidyl peptidase-IV (DPPIV) is a differentiation antigen for human granulosa cells that is initially expressed during corpus luteum formation. To investigate the involvement of cytokines in luteal cell differentiation, we examined the expression and activity of DPPIV in human luteinizing granulosa cells cultured in vitro. Human granulosa cells obtained from patients who had undergone in vitro fertilization were cultured for 7 days in the absence (controls) or presence of hCG (1 U/mL), tumor necrosis factor-alpha (TNF alpha; 10 ng/mL), or interleukin 1-alpha (IL-1 alpha; 10 ng/mL). Flow cytometry showed that the percentage of cultured granulosa cells treated with TNF alpha and IL-1 alpha that was positive for DPPIV expression was significantly higher than that in controls (43.7 +/- 5.4% and 43.4 +/- 5.6%, respectively, vs. 21.7 +/- 3.5%; P < 0.01), whereas hCG treatment produced no remarkable difference in DPPIV expression (24.0 +/- 5.2%). The DPPIV activity of cells treated with TNF alpha and IL-1 alpha was also significantly higher than that of controls, whereas hCG treatment produced no significant difference from control values. These findings indicate that TNF alpha and IL-1 alpha stimulate DPPIV expression and activity in human luteinizing granulosa cells in vitro and suggest the involvement of cytokines in the differentiation of granulosa cells during corpus luteum formation.

Adult

Microtubules are required in amoeba chemotaxis for preferential stabilization of appropriate pseudopods.

Amoebae of Physarum polycephalum exhibit chemotactic responses to glucose and to cAMP. The chemotaxing amoebae exhibit alternating locomotive movements: relatively linear locomotion and movements that change the direction of the locomotion. Such locomotive activity is tightly coupled with the changes in the number and the positions of the pseudopods; cells have one pseudopod at the leading edge during their linear locomotion, while they have multiple pseudopods when they are changing the direction of locomotion. Treatment of cells with microtubule-disrupting reagents inhibited the chemotaxis of the cells. To characterize the role of the microtubule system in chemotaxis, we quantitatively analyzed the relationship between the positions of multiple pseudopods of the amoebae and the relative stability of the pseudopods during reorientation. No significant differences were observed in the pseudopod dynamics between the untreated and the treated amoebae. In both cases, one pseudopod at the leading edge continued to expand during linear locomotion. It then split into two to three pseudopods in the reorientation phase, and the positions of the multiple pseudopods were random. Among multiple pseudopods, however, the pseudopods closer to the microneedle tip were selectively stabilized more often than those distant from the tip in the presence of the microtubule system. By contrast, such preferential stabilization of the appropriate pseudopods was completely abolished by microtubule inhibitors. The microtubule-dependent selection of appropriately located pseudopods enables amoebae to turn correctly at the reorientation step.

Animals

Pharmacological studies on a new dihydrothienopyridine calcium antagonist, S-312-d. 5th communication: anticonvulsant effects in mice.

S-312, S-312-d, but not S-312-l, L-type calcium channel antagonists, showed anticonvulsant effects on the audiogenic tonic convulsions in DBA/2 mice; and their ED50 values were 18.4 (12.8-27.1) mg/kg, p.o. and 15.0 (10.2-23.7) mg/kg, p.o., respectively, while that of flunarizine was 34.0 (26.0-44.8) mg/kg, p.o. Although moderate anticonvulsant effects of S-312-d in higher doses were observed against the clonic convulsions induced by pentylenetetrazole (85 mg/kg, s.c.) or bemegride (40 mg/kg, s.c.), no effects were observed in convulsions induced by N-methyl-D-aspartate, picrotoxin, or electroshock in Slc:ddY mice. S-312-d may be useful in the therapy of certain types of human epilepsy.

Administration, Oral

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

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

[Flare response on bone scintigraphy in metastatic prostate cancer].

Seventy-seven cases of prostate cancer were treated for 5 years at our department and all cases were followed by bone scintigraphy and tumor markers. Of these cases on case of flare response on bone scintigraphy was recognized. A 51-year-old man was hospitalized with chief complaint of lumbago. Serum PAP and gamma-Sm levels were 320 ng/ml and 15 ng/ml, respectively. Prostate biopsy revealed moderately differentiated adenocarcinoma. Bone scintigraphy and CT scan demonstrated multiple bone metastases and lymph nodes involvements. Treatment was started with diethylstilbestrol diphosphate (DES). At one month after the initiation of treatment tumor markers fell down to the normal level and lumbago was diminished, but only serum alkaline phosphatase was elevated and bone scintigraphy showed apparent progression of individual lesions (flare response). The treatment was not altered. At the times after 2, 8, 12 and 36 months successful treatment the bone imaging improved with reduced tracer uptake and no new lesions. The flare response is a healing reaction and is followed apparent improvement. In general, serial bone scintigrams accurately depict the activity of bone metastases in the patients of prostate cancer, but between 1 and 3 months after starting treatment the paradoxical "flare phenomenon" should be taken care.

Adenocarcinoma

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

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

PCNA expression and nucleolar organizer regions in malignant melanoma and nevus cell nevus.

The histogenesis of malignant melanoma with particular reference to the role of melanocytic nevus is still controversial in oncological pathology. In order to differentiate between the proliferative activity of malignant melanomas and benign melanocytic tumors, immunohistochemical analysis was carried out. We used monoclonal antibody PC10 to identify the Proliferating Cell Nuclear Antigen (PCNA), a highly conserved 36 KD acidic nuclear protein which correlates with cell proliferation, and the AgNOR method to stain the Nucleolar Organizer Regions (NORs) whose number and configurations may reflect protein synthesis activity. 47 cases of primary melanoma, 63 metastatic melanoma lesions, and 23 cases of nevi are included in this study. Spitz nevi showed a higher index of cell proliferation compared with other common benign nevi but a much lower index compared with malignant melanoma cells. The metastatic lesions of malignant melanomas had a higher index of proliferation and activity of cells than the primary lesions. Skin-metastatic lesions had higher indexes of cell proliferation and NOR activity than lymph node-metastatic lesions. These results support the idea that metastatic melanoma cells are derived from a more advanced stage of tumor progression. These data suggest that the combination of PCNA and AgNOR is an useful marker for differentiating benign melanocytic tumors from malignant melanomas.

Humans

Intrafamilial outbreak of astrovirus gastroenteritis in São Paulo, Brazil.

This report describes an intrafamilial outbreak of astrovirus-associated gastroenteritis that occurred among 5 children in a family living in the State of São Paul, Brazil. Astrovirus was detected by direct electron microscopy (DEM) and confirmed by solid phase immune electron microscopy (SPIEM) using astrovirus reference serum (T2-6-89). The rise in titre of astrovirus antibody in patients' sera was observed by IEM using acute and convalescent-phase sera. Combined use of the EM techniques to detect virus in stool and antibody rise in sera led us to conclude that this intrafamilial outbreak might have been caused by astrovirus infection. In Brazil, astrovirus may be a more common agent of gastroenteritis than in currently appreciated.

Antibodies, Viral

Bacteriological evaluation of a new air turbine handpiece for preventing cross-contamination in dental procedures.

An autoclavable air turbine handpiece, Air Flushing Clean System (AFCS) (Osada Electric Co., Ltd., Tokyo, Japan) was developed for use in dentistry with the objective of reducing cross-contamination. Its potential for bacterial contamination was investigated in vitro using two bacterial strains (Streptococcus mutants ATCC 25175 and Staphylococcus aureus FDA 209 P). In theory, this device should prevent cross-contamination of the internal water and air lines of the handpiece, by maintaining an internal positive pressure even after the turbine is stopped. In the present study, this AFCS device was found to reduce the bacterial contamination within the air turbine handpiece more effectively than the conventional handpiece used according to accepted protocol. The reduction of such contamination by the AFCS is in keeping with the recent objective of the American Dental Association to reduce cross-contamination during dental procedures.

Bacteriological Techniques

The localization of basic fibroblast growth factor (FGF-2) in rat submandibular glands.

The immunohistochemical localization of basic fibroblast growth factor (FGF-2) in the submandibular glands of the rat was investigated by use of an antiserum to FGF-2. Nerve fiber bundles with FGF-2-immunoreactivity were found in association with interlobular ducts and blood vessels; they dissociated into single immunoreactive nerve fibers perhaps to terminate in proximity to acinar cells, or to form a reticular fiber network within the tunica adventitia of blood vessels. The FGF-2-immunoreactive neurons were located in the submandibular ganglia, but not in the superior cervical ganglia; hence, at least some of these immunoreactive nerve fibers probably come from the submandibular ganglia and are of parasympathetic origin. Most of the epithelial cells of the intercalated and collecting ducts showed notable FGF-2 immunoreactivity. The characteristic distribution of FGF-2 immunoreactivity in both the neuronal and epithelial tissues of the salivary glands suggests a role of this growth factor in complex physiological processes within the salivary glands.

Animals

[Abnormalities and clinical characteristics of growth factors and receptor systems in breast cancer].

Recent progress in molecular biology has revealed that the proliferation of breast cancer is controlled by various growth factors and their receptors. In particular, the amplifications and products of oncogenes which code growth factors and receptors can be different indicators of clinical malignancy from the conventional prognostic factors. We investigated the relation of c-erbB-2 and int-2 gene amplification, expression, ER and EGFR with clinical characteristics. In retrospective study, the cumulative 10-year survival rate of patients with c-erbB-2 and int-2 gene amplification was significantly lower than in patients without amplification. In 72 cases with tumor diameter less than 3.0 cm, which can be an indication of breast preservation surgery, the survival rates of patients in these gene amplified groups were significantly lower than in the non-amplified groups. As for prospective study, the results were the same as those from retrospective study. These data show that the cases with these gene amplification have a high biological malignancy and high risk of recurrence to distant organs, despite the small tumor diameter.

Breast Neoplasms