Search PubMed⌕ Search

Biomedical subjects

M Fujisawa

Publications and source records attributed to M Fujisawa.

At least 253 records · Page 14Linked to original sources
PubMed1986

Recombinant human granulocyte colony-stimulating factor induces granulocytosis in vivo.

Recombinant human granulocyte colony-stimulating factor (G-CSF) was prepared and its granulopoietic effects on mice were examined. When mice were injected intraperitoneally with the G-CSF daily for 14 days, marked granulocytosis occurred in the mice. The number of progenitor cells (CFU-C) was remarkably increased in the spleen. The results suggest that G-CSF plays a central role in granulocyte production in vivo.

Animals↗
PubMed1986

Growth factor(s) produced by a human leukemic cell line growing in a protein-free chemically defined medium.

To examine whether human leukemic cells produce growth factor(s), a protein-free culture line of human erythroleukemic cells (K-562T1) has been established. This unique cell line has been continuously propagated in protein-free Ham's F-10 medium without any supplement for 5 yr. Growth-promoting activity was determined by measuring [3H]thymidine incorporation into DNA in serum-deprived chick embryo fibroblasts. The conditioned medium of K-562T1 contained the growth-promoting activity against chick embryo fibroblasts, mouse 3T3-L1 cells, and K-562 human leukemic cells. This leukemia-derived growth-promoting activity was heat and acid stable and trypsin sensitive. The activity was destroyed by dithiothreitol. Size exclusion chromatography revealed three peaks of activity, with apparent molecular weights of 13,500, 6,300, and 2,400, respectively.

Cell Line↗
PubMed1985

Human myeloid leukemia cells metabolize 25-hydroxyvitamin D3 in vitro.

Human promyelocytic leukemia cells (HL-60) converted 25-hydroxyvitamin D3 to two more polar metabolites during in vitro incubations. A two-step high pressure liquid chromatography system revealed two unique elution positions of those leukemic cell-derived metabolites that exactly co-migrated with the elution positions of 5(Z)-19-nor-10-oxo-25-hydroxyvitamin D3 and 5(E)-19-nor-10-oxo-25-hydroxyvitamin D3, respectively. These unique metabolites did not bind specifically to a protein receptor for 1,25-dihydroxyvitamin D3.

Animals↗
PubMed1985

Activation of the c-K-ras oncogene in a human pancreas carcinoma.

The human pancreas carcinoma cell line T3M-4 contains activated c-Kirsten (K)-ras oncogene detectable by the DNA-mediated gene transfer technique using NIH/3T3 cells. DNA fragments containing coding lesions have been cloned, and nucleotide sequence analysis suggests that the T3M-4 oncogene has been activated by a single nucleotide transition from A to C in the second exon, which results in the substitution of histidine for glutamine in coden 61 of the predicted amino acid sequence. The quantity analysis of c-K-ras oncogene in the DNA and RNA of T3M-4 cells revealed that the c-K-ras gene was amplified and overexpressed in T3M-4 cells. These findings indicate that the T3M-4 c-K-ras oncogene is activated by different mutational events.

Amino Acid Sequence↗
PubMed1985

Increased angiotensin-converting enzyme in peripheral blood monocytes from patients with sarcoidosis.

Angiotensin-converting enzyme (ACE) activity was measured in isolated peripheral blood monocytes and culture medium from 28 patients with sarcoidosis and compared with values obtained from monocytes of 25 normal control subjects. ACE activity was determined by radioimmunoassay of angiotensin II produced from angiotensin I. While there was no measurable ACE activity in monocytes or culture medium from normal controls under the conditions of our study, monocytes from patients with sarcoidosis all showed activity both in cells and culture medium. The mean ACE activity of monocytes from patients with sarcoidosis was 2.0 pg angiotensin II formed/min per 10(5) cells, and that released into medium over a 24-h interval was 30.4 pg angiotensin II/min per 10(5) cells. The monocyte ACE from patients with sarcoidosis was activated by chloride ions and inhibited by EDTA, captopril, and rabbit antiserum to purified human plasma ACE, indicating that enzymatic activity was effected specifically by ACE. Thus, our studies show a significant elevation and release of ACE by peripheral blood monocytes of patients with sarcoidosis under conditions where monocytes of normal control subjects do not demonstrate ACE activity.

Adult↗
PubMed1985

Clinical application of monoclonal antibodies to small cell lung cancer.

Monoclonal antibodies to small cell lung cancer (SCLC) were produced by fusion of P3X63/AgU1 mouse myeloma cells with spleen cells from BALB/c mice immunized against intact cells of SCLC tumors grown in BALB/c athymic nude mice. The antibody TFS-2 demonstrated "pancarcinoma" reactivity showing binding to non-small cell lung cancer (NSCLC) and carcinomas derived from other organs such as stomach, pancreas, and colon. This antibody failed to react with a variety of normal tissues including lung, bone marrow, spleen, stomach, colon, and pancreas. TFS-2 showed complement-mediated cytolysis of target cells. TFS-2 had no significant effects on hematopoietic stem cells. This antibody will serve as a powerful tool for the in vitro removal of tumor cells from bone marrow, thus facilitating high-dose chemotherapy of SCLC with autologous bone marrow transplantation. In contrast, TFS-4 monoclonal antibody reacted specifically with SCLC but not with NSCLS tumors. This antibody can distinguish SCLC from NSCLC tumors in histologic diagnosis at transbronchial biopsy, and in cytological identification of tumor cells in malignant effusion, and in infiltrated bone marrow. Radiolabeled TSF-4 specifically accumulated into SCLC tumors that were implanted subcutaneously into athymic nude mice. Thus, the antibody will be useful not only for diagnosis of lung cancer but also for targeting anti-tumor agents.

Animals↗
PubMed1985

[Transfer of ceftizoxime into serum and prostatic tissue].

Serum and prostatic tissue levels of Ceftizoxime (CZX) were measured in 37 patients with benign prostatic hypertrophy after intravenous infusion of 0.5 g or 1 g or 2 g of CZX. The prostatic tissue level of CZX was sufficient for the treatment of prostatic infection. Serum and prostatic tissue levels of CZX were maintained satisfactorily by the dose of CZX. Neither the weight of extracted prostatic tissue nor the renal function of the patient was correlated to the tissue level of CZX.

Cefotaxime↗
PubMed1985

Familial elevation of serum angiotensin converting enzyme.

We report here a familial clustering of elevated serum angiotensin converting enzyme (ACE) levels. The patient is a 58-year-old Japanese female who had been in excellent health until age 45 when she developed an occlusion of the left central retinal vein. She was otherwise in excellent health, and no laboratory abnormality except a marked elevation of serum ACE level (625 nmol/min/ml; normal range; 22-40 nmol/min/ml of serum) was found. Her blood pressure was within normal limits (140/80 mmHg). There was no evidence for the diagnosis of sarcoidosis, Gaucher's disease, leprosy, hyperthyroidism, diabetic retinopathy, or liver disease. One of her two sisters also showed a marked increase in serum ACE activity (303 nmol/min/ml), and remarkably high levels of serum ACE (276 and 294 nmol/min/ml) were demonstrated in both sons of this sister. All the members of this family have been in excellent health. The serum ACE activity was activated by chloride and cobalt ions, and inhibited by EDTA, captopril and rabbit antiserum to purified human plasma ACE. Thus our study showed a familial clustering of elevated serum ACE in individuals who did not have conventional disease patterns associated with elevated serum ACE.

Angiotensin-Converting Enzyme Inhibitors↗
PubMed1985

Transforming genes in human leukemia cells.

High-molecular weight DNAs of fresh bone marrow cells from 32 patients with fresh leukemia were assayed for the presence of transmissible activated transforming genes by a DNA-mediated gene transfer technique using NIH/3T3 cells. DNAs of bone marrow cells from four of the 32 patients induced transformation of NIH/3T3 cells. Two of the four cases, a chronic myelogenous leukemia and an acute lymphocytic leukemia, contained activated N-ras oncogenes. Molecular cloning and nucleotide sequence analysis revealed that the lesion responsible for the transforming activity was localized to a single nucleotide transition from guanine to thymine in codon 12 of the predicted protein in each of the two cases. These observations indicate that activation of N-ras oncogenes is independent of the specific stage of cell differentiation or the leukemia phenotype. The other two transforming genes associated with an acute myelogenous leukemia and an acute lymphocytic leukemia showed homology neither with members of the ras gene family nor with the human Blym-1 gene. Thus, the NIH/3T3 transfection assay frequently detects activated N-ras oncogenes in human leukemias, while other transforming genes, distinct from the ras gene family, can be detected in some leukemias by the transfection assay.

Autoradiography↗
PubMed1984

Sarcoid granulomas metabolize 25-hydroxyvitamin D3 in vitro.

Sarcoid granulomas metabolized 25-hydroxyvitamin D3 to two unidentified metabolites during in vitro incubation. A two-step high pressure liquid chromatography system revealed two unique elution positions of these sarcoid-derived metabolites that exactly comigrated with the elution positions of 5(Z)-19-nor-10-oxo-25(OH)D3 and 5(E)-19-nor-10-oxo-25(OH)D3, respectively. These unique metabolites did not bind specifically to a protein receptor for 1,25(OH)2D3.

Biopsy↗
PubMed1984

Establishment of a human colony-stimulating-factor-producing cell line from an undifferentiated large cell carcinoma of the lung.

Two different types of lung cancer, a squamous cell carcinoma designated as OTUK, and an undifferentiated large cell carcinoma named as T3M-10, were investigated in vitro; both produced a moderate neutrophilia in each patient. In order to analyze this phenomenon, primary cultures were performed. Epithelial cells attached and grew in both of the cultures. The conditioned medium from both tumor cells revealed high colony-stimulating-factor (CSF) activities. From T3M-10 tumor cultures, a new CSF-producing cell line has been established. T3M-10 cells have been continuously propagated during the last 28 months and produced CSF. Chromosomal analysis revealed the cell line to be a human aneuploid one with a near-diploid mode. These results indicate that the two different types of lung cancers produce CSF, which may have stimulated granulopoiesis in vivo in the patients as well.

Adult↗
PubMed1984

Long-term cultivation and differentiation of human erythroleukemia cells in a protein-free chemically defined medium.

To examine whether a human erythroleukemia cell line, K-562, can proliferate and differentiate without protein supplements, long-term cultivation of the cells was carried out in a protein-free chemically defined medium. By the use of stepwise decreases in the fetal bovine serum concentration, continuous growth of K-562 cells was established in a protein-free F-10 medium. The cells have been propagated in this medium for 3 years. The population-doubling time of the cells is about 30 hr. Growth was not stimulated by the addition of insulin, epidermal growth factor, fibroblast growth factor, multiplication-stimulating activity, transferrin, platelet-derived growth factor, or dexamethasone. Addition of serum stimulated the cell growth slightly and increased the saturation density. The saturation density of the cells could be increased to that seen with serum-supplemented cultures by changing the serum-free medium daily. The cells synthesized significant amounts of hemoglobin in the presence of hemin without serum supplementation. The results suggest that the human erythroleukemia cells grown in a protein-free medium do not require serum components for their growth or hemin-induced hemoglobin synthesis and provide an excellent model for better understanding of the growth and differentiation of human leukemia cells.

Butyrates↗
PubMed1984

Monoclonal antibodies to surface antigens of small cell carcinoma of the lung.

Monoclonal antibodies to membrane antigens of human small cell carcinoma of the lung were produced by fusion of P3X63/Ag8U1 mouse myeloma cells with spleen cells from BALB/c mice immunized against the intact cells of the small cell carcinomas grown in BALB/c nude mice. The hybrids were screened for antibody production using intact cells in a solid-phase radioimmunoassay or in a membrane fluorescence with a fluorescence-activated cell sorter. Four monoclonal antibodies were chosen that demonstrated reactivities with human small cell carcinoma of the lung and not with apparently normal diploid fibroblasts or lymphoblastoid cells. The antibodies designated as TFS-1 and TFS-2 rather demonstrated "pancarcinoma" reactivity, showing binding to the other types of lung cancer (adenocarcinoma, squamous cell carcinoma, and large cell carcinoma) and carcinomas derived from other organs, such as colon, pancreas, or stomach. The monoclonal antibodies TFS-3 and TFS-4 preferentially bound to small cell carcinoma cells and neuroblastoma cells, but not to non-small cell carcinomas (adenocarcinoma, squamous cell, or large cell). Especially, TFS-4 did not bind to a variety of other normal or malignant cells. Immunoprecipitation of the antigens by monoclonal antibodies and sodium dodecyl sulfate:polyacrylamide gel electrophoresis revealed that they had different molecular weights.

Animals↗