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Biomedical subjects

R Ueda

Publications and source records attributed to R Ueda.

At least 289 records · Page 16Linked to original sources
PubMed1990

Point mutations of c-ras genes in human bladder cancer and kidney cancer.

Point mutations of c-ras genes at codons 12, 13 and 61 were analyzed in 26 cases of bladder cancer and 16 cases of kidney cancer. DNA prepared from either frozen tissues or 10% formalin-fixed, paraffin-embedded tissues were amplified by means of polymerase chain reaction methods, and mutations were analyzed by dot blot hybridization assays with oligonucleotide probes. In three cases of bladder cancer c-ras mutations were found, at codons 13 and 61 of c-Ha-ras and at codon 61 of c-Ki-ras, while no mutation was found in kidney cancer. No mutation was found in normal bladder epithelial tissues from the same patients. Our findings, taken together, may indicate relative scarcity of c-ras mutations in these types of human cancer. The results of dot blot hybridization assays and DNA sequencing showed a G-to-C transition of the first nucleotide at codon 13 c-Ha-ras. This is the first time that such a point mutation has been detected in human cancer tissues.

Aged↗
PubMed1990

Long-term follow-up study of cerebral palsy children and coping behaviour of parents.

The purpose of this study was to understand: (a) the feelings, thoughts and actions of parents at the time their children were diagnosed as having cerebral palsy; (b) the crisis periods in raising their children; (c) the important persons who supported the parents during the 'acceptance' phase; and (d) the roles of mothers and fathers in raising the children. The subjects were 28 mothers and fathers who had sons or daughters with cerebral palsy. The latter offspring were aged 22-29 years at the time of this study. They were interviewed at their homes with a semi-structured method retrospectively. The results showed that most parents became aware of their children's disability in infancy and most of their children were diagnosed as having cerebral palsy by around 2 years of age. The mothers' reactions to the diagnosis were emotional and those of the fathers were realistic in coping with the problem. Although crisis periods arose throughout the periods of growth and development of the children, the infancy period was the more critical for mothers and the toddlerhood, school-age and adolescence periods were more critical for fathers. The important support people were the spouses. The mothers took care of the children while the fathers provided an income; however, some fathers withdrew from competing for achievement in their jobs.

Adaptation, Psychological↗
PubMed1990

Assay for the soluble interleukin-2 receptor by sandwich enzyme linked immunosorbent assay.

A system for the detection of soluble IL-2R by sandwich enzyme linked immunosorbent assay (ELISA) was established. This assay system has good reproducibility, and was found to be specific for soluble IL-2R by examination of the binding of anti-IgM antibody or anti-IL-2R monoclonal antibody (mAb) with IgM or soluble IL-2R and inhibition by IL-2 of the binding of anti-IL-2R mAb to soluble IL-2R. The IL-2R molecules in supernatant of phytohemagglutinin-stimulated peripheral blood mononuclear cells were of 44,000-61,000 molecular weight as estimated by size of exclusion high performance liquid chromatography. The results of determinations for soluble IL-2R with a Eurogenetics kit (sandwich enzyme linked immunosorbent assay) were significantly correlated with those by the our method.

Enzyme-Linked Immunosorbent Assay↗
PubMed1990

In vitro and in vivo properties of human/mouse chimeric monoclonal antibody specific for common acute lymphocytic leukemia antigen.

A human/mouse chimeric monoclonal antibody specific for a common acute lymphocytic leukemia antigen was efficiently obtained by ligating human heavy-chain enhancer element to the chimeric heavy- and light-chain genes. Cell binding and competitive inhibition assays of both radioiodine and indium-111- (111In) labeled chimeric antibodies demonstrated in vitro immunoreactivity identical with that of the parental murine monoclonal antibodies. The biodistribution of the radiolabeled chimeric antibody in tumor-bearing nude mice was similar to that of the parental murine antibody. Tumor accumulation of radioiodinated parental and chimeric antibodies was lower than that of 111In-labeled antibodies, probably because of dehalogenation of the radioiodinated antibodies. Indium-111-labeled chimeric antibody clearly visualized xenografted tumor. These results suggest that a human/mouse chimeric antibody can be labeled with 111In and radioiodine without the loss of its immunoreactivity, and that chimeric antibody localizes in vivo in the same way as the parental murine antibody.

Animals↗
PubMed1989

Chemosensitivity and radiosensitivity of small cell lung cancer cell lines studied by a newly developed 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) hybrid assay.

The 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) hybrid assay was developed by technically combining the human tumor clonogenic assay and the MTT assay to make the most of both assays. This assay was able to estimate the in vitro growth of cultured cell lines and of tumor cells in pleural effusion, suggesting the possibility of its use for assessment of chemosensitivity and radiosensitivity of fresh tumor samples. Multiple cell lines [including morphological and/or phenotypical in vitro converters and cisplatin (CDDP)-resistant lines] were established from three patients with small cell lung cancer at different stages of the disease. Chemosensitivity of these cell lines to four commonly used chemotherapeutic drugs was tested by the MTT hybrid assay. SK1 and SK3 lines were established from Patient S. K. before and after chemotherapy and radiotherapy, respectively. SK3/CDDP, a CDDP-resistant line derived from the SK3 line, was 30-fold more resistant to CDDP [50% inhibiting dose (IC50), 21.5 micrograms/ml] than the SK1 line. In Patient M. O., MOA2/CDDP, a CDDP-resistant line derived from MOA2 (an in vitro converter from the MO line), was 41-fold more resistant to CDDP (IC50, 37 micrograms/ml) than the parent MO line. From Patient T. M., TM1 and TM2 lines were established before and after chemotherapy, respectively. The latter showed 6-fold more resistance to CDDP than the former. Chemosensitivity of these lines to three other drugs, 4-hydroperoxycyclophosphamide, Adriamycin, and etoposide, suggested cross-resistance between CDDP and 4-hydroperoxycyclophosphamide. Radiosensitivity study was also carried out with the MTT hybrid assay. The MOA2 line was more resistant [Do, 3.0 Gy; extrapolation number (n), 4.0] than the parental MO line (Do, 1.6 Gy; n, 2.1). There was no clear difference in radiosensitivity between the cell lines established before and after radiation therapy in Patient S. K.

Antineoplastic Agents↗
PubMed1989

Elevated serum-soluble interleukin-2 receptor (Tac antigen) levels in chronic myelogenous leukemia patients with blastic crisis.

We examined the expression of cell-surface interleukin-2 (IL-2) receptor (Tac antigen) on peripheral blood leukemic cells and measured soluble IL-2 receptor p55(alpha) chain (sIL-2R) levels in sera from chronic myelogenous leukemia (CML) patients with blastic crisis. Flow cytofluorometric analysis performed by dual immunofluorescence in three cases demonstrated coexpression of Tac antigen with myeloid (CD13, CD14, or CD33) or lymphoid (CD10) antigen on significant proportions of peripheral blood leukemic cells. Radiolabeled IL-2-binding assay demonstrated the specific IL-2 binding sites in three cases examined. The exogenous IL-2, however, failed to induce proliferative response. A myeloid cell line, Yut-K3, established from peripheral blood leukemic cells from a CML patient with blastic crisis, also expressed cell-surface Tac antigen and CD13 concurrently. SIL-2R assay showed that Yut-K3 released a detectable amount of sIL-2R in its culture supernatant. The serum sIL-2R levels were significantly elevated (range: 2,580 to 172,000 U/mL) in 12 CML patients with blastic crisis and were slightly elevated in ten patients in chronic phase (range: 250 to 820 U/mL) and in three in accelerated phase (range: 790 to 1,305 U/mL) compared with those in 24 normal controls (range: 70 to 695 U/mL, P less than .01). These results indicated that the leukemic cells from CML patients with blastic crisis expressed and released IL-2 receptor (Tac antigen). Longitudinal studies performed in three cases of CML with blastic crisis showed that the change of serum sIL-2R level was closely associated with that of the number of peripheral blood leukocytes and blasts, the percentage of blasts and serum LDH levels, also suggesting that the serum sIL-2R level is a useful clinical indicator of the leukemic cell burden in vivo.

Adult↗
PubMed1989

Expression and amplification of myc gene family in small cell lung cancer and its relation to biological characteristics.

Eighteen small cell lung cancer (SCLC) lines (including nine lines established by this group) as well as 31 tumor samples from 23 SCLC patients were examined for the surface antigen phenotype and the expression and amplification of the myc gene family. The expression of NE-150 neuroendocrine, PE-35 panepithelial and OE-130 epithelial antigens corresponded well with the level of biomarkers of SCLC lines, i.e., the NE-150+/PE-35+/OE-130- phenotype corresponded to classic type, while the other phenotypes such as NE-150+/PE-35-/OE-130- to variant type. In tumor specimens, most classic SCLC (consisting of oat cell type and intermediate cell type, subtype a) showed NE-150+/PE-35+/OE-130- phenotype, while small cell-large cell carcinoma (intermediate cell type, subtype b) expressed various phenotypes. The amplification of the myc gene family was observed in nine out of 18 lines (50%) and five out of 23 patient tumors (22%). Higher levels of expression of either c-myc, N-myc, or L-myc were detected in 16 out of 18 lines (89%) and in five out of six patient tumors (83%), when compared with that of normal or fetal lung tissues. Thus, the higher expression without obvious myc gene amplification was observed. The cell lines and tumors with the amplified myc always expressed their corresponding myc genes. The results suggested that higher levels of expression of the myc gene family may play a significant role in the oncogenesis of SCLC. Amplification and/or high levels of expression of c-myc were observed not only in variant type SCLC lines, but also in classic type lines. Thus, they were not necessarily associated with distinct biomarkers of SCLC lines.

Antigens, Surface↗
PubMed1989

Interdigitating cell sarcoma (ICS). Evidence of interdigitating cell origin, immunocytochemical studies with monoclonal anti-ICS antibodies.

Three independent mouse monoclonal antibodies (mAbs) ID1 (IgG3), ID2 and ID3 (IgM) were raised against whole cells of a surgically resected human interdigitating cell sarcoma (ICS). In immunoperoxidase staining, these mAbs strongly stained the cytoplasm of ICS neoplastic cells as well as interdigitating cells in normal lymphoid tissues. These mAbs also detected monocyte/macrophages and dendritic cells, although their staining was highly variable depending on tissue distribution of the cells. Additional immuno-histological and enzyme histochemical study revealed that the neoplastic cells of ICS had cytoplasmic acid phosphatase and membranous alkaline phosphatase activity, and also possessed S100 beta protein, Ki-1 antigen. DAKO-macrophage antigen, and weak vimentin activity. Neither rearrangement of immunoglobulin heavy chain gene nor of T-cell receptor genes was detected in the DNA of ICS by Southern hybridization. These observations provide further confirmation of our previous finding (Nakamura et al. 1988, 1989) that the origin of ICS is interdigitating rather than lymphoid cell, and indicate that our mAbs could be useful as a cellular differentiation marker of interdigitating cells and for diagnosis of ICS.

Animals↗
PubMed1989

A human monoclonal antibody recognizing a surface antigen on stomach cancer cells.

Lymph-node lymphocytes of a patient with stomach cancer were fused with the mouse-human heterohybridoma, HM-5. A clone (2F9) was isolated that showed stable production of an IgM antibody reactive with NUGC-4 stomach cancer cell line. This antibody reacted predominantly with a cell surface antigen on cell lines originating from gastro-intestinal cancer and adenocarcinoma of lung, whereas it was not generally reactive with other types of cancers, or with normal kidney cells or fibroblasts. Biotin-labeled 2F9 antibody clearly stained cell smears and the nude mouse tumor of NUGC-4, but it did not show a positive reaction with stomach cancer tissues obtained from more than 10 patients, indicating that the antigen detected is very weakly expressed on tumor cells or on a limited number of stomach cancers. The antigen shed from NUGC-4 cell line was detected in the culture supernatant. 2F9 antibody precipitated a glycoprotein with a molecular weight of over 200 kilodaltons as well as a possible glycolipid, from NUGC-4 cells labeled with [3H]glucosamine or [35S]-H2SO4. Periodic acid treatment of the tissue section decreased reactivity with 2F9 antibody, but heat, neuraminidase or protease treatment did not. These results suggested that the epitope is present on a carbohydrate moiety not containing sialic acid, and that a part of the antigen molecule is sulfated.

Animals↗
PubMed1989

bcl-2 gene rearrangement analysis in Japanese B cell lymphoma; novel bcl-2 recombination with immunoglobulin kappa chain gene.

The rearrangement of bcl-2 gene was studied in 56 Japanese B cell lymphoma cases to investigate the contribution of bcl-2 gene to lymphomagenesis in Japan. Ten out of 56 cases showed bcl-2 gene rearrangement; it was detected in only 5 out of 16 follicular lymphoma cases (31%) and in 5 out of 40 diffuse B cell lymphoma cases (13%). The incidence of bcl-2 gene involvement in Japanese follicular lymphomas was lower than those reported in the United States. This might contribute to the lower incidence of follicular lymphoma cases in Japan. Novel recombination between bcl-2 and Ig kappa genes at the 5' region of bcl-2 and J kappa 4 segment was observed in one follicular lymphoma case, suggesting that bcl-2 gene is transcriptionally activated by Ig kappa enhancer. It was also suggested that this case had originated from a more differentiated B cell than most follicular lymphomas with bcl-2-Ig H recombination.

B-Lymphocytes↗
PubMed1989

Identification of myeloid origin in undifferentiated congenital leukemia by in vitro marrow culture study.

A case of congenital leukemia that originally did not express any lineage-specific antigenic markers is presented. The blast cell morphologic appearance was L1 according to French-American-British (FAB) classification and showed lymphoid characteristics by cytochemical staining and transmission electron microscopy. However, immunophenotyping using a variety of monoclonal antibodies did not confirm the lymphoid origin. Furthermore, the immunoglobulin heavy chain and T-cell receptor beta-chain genes were in germ-line configuration. The in vitro culture study defined the leukemia as of myeloid origin. The semisolid methylcellulose culture showed an acute non-lymphocytic leukemia-type growth pattern. Bone marrow blasts underwent myeloid differentiation with positive myeloperoxidase and butyrate esterase activity during a suspension culture. These findings indicate that this case represents an acute undifferentiated leukemia that has probably arisen from the malignant transformation of stem cells of myeloid progeny.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed1989

Blastogenic responses of human lymphocytes to a streptococcal preparation OK-432 and its fractions.

This study was undertaken to examine the blastogenic responses of PBL to Streptococcal preparation OK-432 and its fractions. The results showed that OK-432 and its fractions, Su-PR and PM, have the mitogenic activity against human lymphocytes. The demonstration that PBL which had been cultured with OK-432 expressed IL-2 receptors and the blastogenic responses of PBL to OK-432 were inhibited by anti-IL-2 antibody suggested that IL-2 and IL-2 receptors might play a central role in OK-432-induced proliferation of human lymphocytes.

Adolescent↗