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Biomedical subjects

Y Tamaki

Publications and source records attributed to Y Tamaki.

At least 19 recordsLinked to original sources

NMDA-receptor mediated electrical epileptogenesis in the organotypic culture of rat hippocampus.

Extracellular field recordings were made in CA1 in the hippocampal explant cultures in oxygenated artificial cerebrospinal fluid. Schaffer collaterals were stimulated with 1-s trains of 60 Hz pulses every 10 min. Seizures were reliably elicited with progressive lengthening over 1-2 h. D-APV, an N-methyl-D-aspartate (NMDA) antagonist, stereoselectively blocked the development of seizures. Thus we have demonstrated that in vitro epileptogenesis occurs in hippocampal explant cultures through NMDA receptor mediated mechanisms.

2-Amino-5-phosphonovalerate

A human monoclonal antibody derived from axillary lymph nodes of a breast cancer patient reactive to a sulfated glycolipid.

A human monoclonal antibody, BMMK-33G, was established by a fusion of human B-lymphoblastoid cells, HO-323, with lymphocytes of axillary lymph nodes obtained from a breast cancer patient. High-performance thin-layer chromatography (HPTLC)-immunostaining and enzyme-linked immunosorbent assay (ELISA) revealed that BMMK-33G was interestingly directed to enough sulfatide (Galactosylceramid-I2-sulfate), which is one of the sulfate ester containing glycolipids. By immunohistochemical staining, BMMK-33G intensely reacted to breast cancer, pancreatic cancer and gastric cancer. It also reacted to many normal human tissues including mammary glands, but these stainings were weaker than those for cancer. This report describes BMMK-33G, a human monoclonal antibody against sulfatide which may be very useful for studying not only tumor immunology but also autoimmune diseases.

Antibodies, Monoclonal

Use of a local immunotherapy as an adjunctive tool for the generation of human monoclonal antibodies from regional lymph nodes of colonic cancer patients.

Human hybridomas were generated through the fusion of the human B-lymphoblastoid cell line HO-323 with the regional lymph node lymphocytes of colonic cancer patients who had received a local immunotherapy. A total of 353 hybridomas were obtained from 4 patients and 116 of these were found to secrete greater than or equal to 100 ng/ml human immunoglobulin. The efficiency was remarkably high as compared with that from patients without the local immunotherapy. Further immunohistological examination showed that 5 hybridomas secreted IgM which selectively reacted with colonic cancers. The results indicate that local immunotherapy could be an adjunctive tool for the generation of highly potent human hybridomas through augmenting the host's immunity.

Antibodies, Monoclonal

Malignant lymphomas of the nasal cavity and paranasal sinuses. A clinicopathologic and immunohistochemical study.

The clinicopathologic and immunohistological features of 20 Japanese patients with non-Hodgkin's lymphomas (NHLs) limited to the sinonasal area were studied using a broad panel of T- and B-cell markers on paraffin-embedded and fresh frozen tissue. All cases showed a diffuse growth pattern. Nine cases were B-cell lymphomas (immunoblastic n = 4, centroblastic n = 3, immunocytoma n = 1, centrocytic n = 1), and nine were T-cell lymphomas (pleomorphic medium and large cell n = 8, angioimmunoblastic n = 1). In two cases, the cell lineage could not be determined. No morphologic features of angiocentric/angiodestructive lymphoproliferative lesions or lymphoepithelial lesions in ductal or glandular epithelium were seen in our series. Eight (89%) of the nine T-cell tumors and four (44%) of the nine B-cell neoplasms involved both the nasal cavity and paranasal sinuses. Six of the nine T-cell neoplasms showed a clinical presentation of rhinitis, whereas all of the B-cell neoplasms showed tumor masses in the nasal cavity and/or paranasal sinuses. The two-year survival rate for T-cell lymphomas was poorer than that for B-cell lymphomas. The five-year survival of patients with NHLs involving both the nasal cavity and paranasal sinuses was also poorer than that of patients in whom NHLs were limited to the nasal cavity.

Adult

Efficient methods for production of monoclonal antibodies for forensic applications.

To simplify the labor-intensive process of making hybridomas, we fused popliteal lymph node cells, used an autoclavable serum-free culture medium throughout, and cloned hybridomas with human blood cell feeders. To make the best possible use of the serum-free medium, we adapted mouse myeloma cells to it. Using as little as 0.2 ml of human blood per culture plate, we successfully cloned hybridomas and established a hybrid cell line producing anti-peroxidase antibody. Our protocol affords a considerable saving on time, labor, and cost, and hopefully will encourage the forensic scientist to undertake the production of monoclonal antibodies.

Animals

[A case report of metastatic colonic carcinoma in the liver effectively treated by long-term, ambulatory and continuous, intra-hepatic-arterial infusion of 5-fluorouracil using disposable multi-day-type infusor].

A 42-year-old female was diagnosed as having sigmoid colonic carcinoma with multiple metastases in the liver. Following sigmoid colectomy and descending colostomy, a catheter was inserted from the right gastroepiploic artery to the proper hepatic artery. From the day of surgery 5-Fluorouracil was administered in doses of 250 mg/day continuously through a catheter over the 2-month period of hospitalization. After the patient was discharged, 250 mg/day of 5-Fluorouracil was administered at home using Vaxter Infusor according to a regimen of 10-day continuous infusion and subsequent 4-day rest. Five months after the initial operation, the serum CEA level decreased dramatically, and CT scan of the liver revealed the complete disappearance of the metastases. The patient underwent a second operation in which the colostomy was closed, and she is doing well at this writing. This case suggests that long-term, ambulatory, continuous and intra-hepatic-arterial infusion of 5-Fluorouracil can be a very effective treatment not only in reducing the hepatic metastases but also in improving the quality of life of patients with colonic carcinoma.

Adult

Production and characterization of a monoclonal antibody to ABH-carrying alpha 2-seminoglycoprotein for ABO grouping of semen by ELISA.

BALB/c mice were immunized with alpha 2-seminoglycoprotein (A2SGP), the lymph node cells were fused with P3U1 myeloma cells and cultured by the conventional technique. Four antibody-producing hybridoma clones were established and antibody-containing ascitic fluid obtained. The antibody was directed to the protein backbone of A2SGP and not to ABH antigenic determinants and did not cross-react with saliva or vaginal secretions. When tested in an indirect ELISA the anti-A2SGP antibody had a titer of 512000. The anti-A2SGP was used in a capture ELISA (or sandwich ELISA) in which wells were coated with this antibody to capture A2SGP in semen, and the captured A2SGP was detected with anti-A, anti-B or anti-H-peroxidase conjugate and peroxidase-labeled second antibody. This ELISA allowed correct ABO grouping even of 1:12,800 or higher dilutions of semen. When the ELISA was applied to ABO grouping of seminal fluids mixed with vaginal secretions only the seminal ABH antigens could be detected. The results strongly suggest the potential usefulness of monoclonal anti-A2SGP in the investigation of rape cases.

ABO Blood-Group System

A Japanese family with unusual segregation of GM phenotype: a GM silent allele?

A Japanese family with unusual segregation of GM phenotype was found in connection with a case of disputed paternity. Typing for 27 hemogenetic marker systems revealed apparent nonmaternity in the GM system alone. When tested for 14 GM allotypes, the alleged father, the mother, and the child were of the GM A,F,Z N B0,1,3,4,5,S,T,U; GM A,Z N' G,U; and GM A,Z N' B0,3,5,S,T types, respectively. The results of a family study strongly suggested that a GM silent allele was responsible for the apparent nonmaternity; however, definitive evidence could not be obtained.

Adult

Atypical inheritance of hemogenetic markers: seven cases of disputed parentage.

The present paper reports seven families with an atypical segregation pattern which we have recently found during paternity testing. In one family, heterozygous mother-child incompatibility (1B2A and 1A2B) in the PGM1 system was observed. This could be explained by an intragenic recombination between the PGM1*1B and PBM1*2A alleles giving rise to the PGM1*2B allele. In the other families, two types of new GC variants with two pairs of double band were identified. These variants (GC*1S1A2 and GC*1F1A2) were presumed to have arisen from the duplication of two GC*1 alleles.

Crossing Over, Genetic

HTLV-I-associated myelopathy (HAM/TSP) with Hashimoto's thyroiditis.

Two cases of HTLV-I-associated myelopathy (HAM/TSP) with Hashimoto's thyroiditis are reported. One patient is a 47-year-old female with goiter and the other a 60-year-old female. Both have slowly progressive spastic paraparesis and have anti-HTLV-I antibodies in their serum and cerebrospinal fluid. Anti-thyroid antibodies are also present in their serum, and biopsy specimens of their thyroids showed the pathological features of Hashimoto's thyroiditis. The association of the two disorders in these patients provides further evidence for the autoimmune hypothesis of the pathogenesis of HAM/TSP, and also suggests a relationship of Hashimoto's thyroiditis with HTLV-I infection.

Autoimmunity

Practical method for production of monoclonal antibody to human IgG allotype G1M F and its applications in ELISAs and dot immunobinding.

An anti-G1M F monoclonal antibody was produced by immunization of mice with a single dose of F(ab')2 fragments of normal IgG1-enriched IgG and subsequent fusion of their lymph node cells with P3U1 myeloma cells. Antibody specificity was tested by an ELISA in microtiter plates coated with allotype positive or negative IgG. The usefulness of the antibody as a G1M F typing reagent in inhibition and direct immobilization-type ELISAs and dot immunobinding was demonstrated by re-typing of 100 GM-allotyping control sera. The advantages and disadvantages of these assay methods are discussed.

Animals

Experimental glycerol myopathy: a histological study.

Histopathological changes induced by the intramuscular injection of glycerol were studied in the muscle fibers of rabbits. Fifteen minutes after the injection of 1 ml of 50% (v/v) glycerol, hypercontraction of fibers, disruption of the plasma membrane, and invasion of lanthanum into the sarcoplasm were observed. Between 12 and 24 h after the injection, more extensive pathological changes were seen which included: variation in fiber size; degeneration and necrosis of muscle fibers; hypercontraction of fibers by light microscopy; disruption of the plasma membrane by electron microscopy; vacuolar changes; hypercontraction of myofibrils, and selective loss of Z-bands. Between 7 and 14 days after glycerol injection, extensive regenerative changes were seen. The degenerative changes were similar to those seen in muscle in Duchenne muscular dystrophy, suggesting that a similar mechanism may be involved in the two conditions, so that experimental glycerol myopathy could be a good model for pathophysiological studies on Duchenne muscular dystrophy.

Acid Phosphatase

G3M T typing by dot immunobinding with monoclonal anti-G3M T antibody.

Dilutions of 100 serum samples of various GM phenotypes were dotted onto a nitrocellulose membrane. The serum dot-blots were detected with peroxidase-labeled anti-G3M T monoclonal antibody (anti-G3M T MAb). Up to a 1 : 256 dilution could be G3M T-typed correctly. By use of anti-G3M T MAb and peroxidase-labeled anti-mouse IgG or the biotin-avidin system instead of use of labeled anti-G3M T MAb, up to a 1 : 512 or 1 : 1024, respectively, dilution was typable. As with previous work with G3M G MAb, the dot immunobinding (DIB) method for G3M T typing was found very simple and practical.

Antibodies, Monoclonal

[Automatic polishing of dental prostheses. 1. Development of a centrifugal barrel finishing apparatus].

A centrifugal barrel finishing apparatus with a variable turn table rotational speed (250-1,000 rpm) was newly developed and barrel finishing of Ni-Cr casting plates (10 x 10 x 2 mm) was performed using alumina base chips. When using the sample with a mirror face the amount of polishing and the surface roughness increased and the surface gloss decreased with the increase in rotation speed. A high rotational speed was useful for coarse polishing and low rotational speed was useful for fine polishing. The continuous barrel polishing was trially performed using the sample prepared by the carborundum wheel under variable rotational speed. Automatic polishing to fine polishing could be done using this apparatus.

Dental Polishing

Production of monoclonal antibody to human IgG allotype G3M T.

An efficient method for the production of monoclonal anti-G3M T antibody is described. IgG3 protein of GM B3ST phenotype was isolated by affinity chromatography on Ricinus communis lectin I-agarose and used for immunization. A mouse hybridoma clone was obtained by fusion of popliteal lymph node cells and P3U1 myeloma cells. The antibody produced was tested for allotype specificity by hemagglutination inhibition and ELISA methods using 101 IgG-allotyping control sera. The antibody was neutralizable by all G3M T-positive sera and entirely nonneutralizable by G3M T-negative sera in the inhibition test, and reacted only with G3M T-positive IgG coats in the ELISA test. The results prove the antibody to be allotype-specific, and therefore practically establishes its monoclonality.

Antibodies, Monoclonal

Effects of peripheral chemo- and baro-receptor denervation on responses of preoptic thermosensitive neurons to inspired CO2.

The purpose of this study was to see if the responses of thermosensitive neurons in the preoptic (PO) area to inspired CO2 seen in spontaneously ventilated rats were indirectly driven by reflexive changes in respiration and circulation. In urethanized, paralyzed, and artificially ventilated (AV) rats, the effects of 10% CO2 inhalation on PO thermosensitive neurons were examined by regression of neuronal activity on PO temperature. The experiments were made in intact rats and in rats whose peripheral chemo- and baro-receptors were denervated (AVD). In both AV and AVD rats, the slope of the regression line decreased significantly (P less than 0.05) during CO2 inhalation in half of the warm-sensitive neurons studied (64.3% in AV rats, 41.7% in AVD rats). Peripheral chemo- and baro-receptors thus do not appear to be responsible for decreased thermosensitivities of warm-sensitive neurons during CO2 inhalation. The tendency for activities of warm-sensitive neurons to increase progressively at lower Tpo was seen during CO2 inhalation in both AV and AVD rats. However, the average differences in mean firing rate between 10% CO2 and air inhalations were 2-3 imp/s greater at any Tpo in AV rats than in AVD rats. In AVD rats, warm-sensitive neurons were rather inhibited by CO2 at higher Tpo. Excitation of warm-sensitive neurons during CO2 inhalation in AV rats, which was independent of Tpo, was considered to be caused by the signals from peripheral receptors.

Administration, Inhalation

A human monoclonal antibody derived from axillary lymph nodes of a breast cancer patient.

A human hybridoma clone (A4-33) was established by fusion of human lymphoblastoid cells, HO-323, with lymphocytes of axillary lymph nodes obtained from a breast cancer patient. This clone has been stable, producing IgM for over 24 months. Enzyme-linked immunosorbent assay (ELISA) showed that monoclonal antibody A4-33 reacted strongly to MCF-7, and weakly to PANC-1 and HT-29, but not to other human malignant cell lines. The reactivity of A4-33 to MCF-7 was markedly reduced by treatment with periodate, but not affected or enhanced by neuraminidase or trypsin. Immunoperoxidase staining of normal human tissue sections showed that A4-33 reacted to acinar and ductal cells of the mammary gland and to ductal epithelial cells of the salivary gland, sweat gland, pancreas and bile duct. With malignant tumor sections, A4-33 reacted to breast cancer, pancreatic cancer and parotid cancer. These results suggested that A4-33 recognized the antigen which commonly exists on ductal epithelial cells of the exocrine gland.

Antibodies, Monoclonal