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Biomedical subjects

S Asano

Publications and source records attributed to S Asano.

At least 433 records · Page 24Linked to original sources
PubMed1988

Protective effect of human granulocyte colony-stimulating factor on bacterial and fungal infections in neutropenic mice.

The in vivo effects of purified natural human granulocyte colony-stimulating factor (h G-CSF) on bacterial and fungal infections were examined in mice treated with cyclophosphamide (CPA). Induction of severe neutropenia was prevented by repeated daily injections of h G-CSF at a dose of 2.5 micrograms/day for 4 days after the CPA treatment. Although control mice died shortly after the bacteria inoculation at day 4, most of the mice in whom the numbers of blood neutrophils were restored by h G-CSF injections were saved from death. At the site of the bacteria inoculation in the h G-CSF-treated mice many mature neutrophils were observed to migrate in a dose-dependent way. Furthermore, the dose of antibiotics required for elimination of infections was proved to be much less in the h G-CSF treated mice than that in the non-treated mice. These findings indicate that h G-CSF can be used efficiently for prevention and treatment of bacterial and fungal infections, particularly in patients with neutropenia.

Agranulocytosis↗
PubMed1988

Phase I clinical study for recombinant human granulocyte colony-stimulating factor.

The pharmacokinetics, the safety and the efficacy of purified recombinant human granulocyte colony-stimulating factor (rh G-CSF) expressed in Chinese hamster ovary (CHO) cells were studied in normal healthy adults. Following the single subcutaneous dose, G-CSF levels in sera elevated in a dose-dependent way, with C-max of approximately 140 pg per ml, T-max of 4 hours and T-half of 5 hours at a dose of 10 micrograms per ml. In accord with the elevation, blood neutrophil counts rose promptly and transiently without any changes in the other blood cell counts. Degree and duration of the neutrophilia were apparently proportional to the rh G-CSF dose and, in turn, the G-CSF levels in sera. Single daily injections of rh G-CSF at a dose of 10 micrograms per body per day for 7 consecutive days did not bring on any accumulation of rh G-CSF in sera, and repeatedly induced neutrophilias following every injection. During the course of the injections, the chemiluminescence generating ability of blood neutrophils also rose. Peak value of the transient neutrophilias and level of the abilities tended to go down after 3 or 4 consecutive injections, presumably due to the limited number of mature neutrophils which can be released from the bone marrow in response to rh G-CSF. However, it was found that the M/E ratio becomes higher after the repeated dosing with an increased proportion of myeloid immature cells as well as increase in the levels of both erythroid and myeloid precursor cells in the bone marrow.(ABSTRACT TRUNCATED AT 250 WORDS)

Colony-Forming Units Assay↗
PubMed1988

[Clinical studies of natural interferon alpha (HLBI) in chronic myelogenous leukemia--a multi-institutional cooperative study in Japan].

Clinical trials of natural interferon-alpha (HLBI) in the treatment of Philadelphia chromosome (Ph1) positive chronic myelogenous leukemia (CML) were carried out in a cooperative study of 22 institutions in Japan. Patients with CML in chronic and accelerated phase were given intramuscular or subcutaneous injections of HLBI at the dose of 6 X 10(6) U/body on consecutive days. Of the 47 patients in this study, forty-one were evaluable for clinical effects, and 42 could be assessed for adverse effects. Among 30 evaluable patients in the chronic phase, 9 (30.0%) achieved complete remission (CR) and 20 (66.7%) partial remission (PR). Only one patient had no response (NR), and the response rate (CR + PR) in chronic phase was 96.7% (29/30). Among 11 patients in the accelerated phase, 3 (27.3%) achieved CR and 4 (36.4%) PR. The response rate in the accelerated phase was 63.6% (7/11). In all 41 evaluable patients, the response rate was 87.8% (36/41). In 5 of 13 responding patients treated for more than 6 months, cytogenetic investigation showed the decline of Ph1 positive bone marrow cells from 100% to 92-0% (mean 46%). Adverse effects such as fever (52.4%), general fatigue (35.7%), and liver dysfunction (21.4%), were observed, but they were usually mild and reversible. No patient was taken off this study because of these toxicities. The results confirm the clinical efficacy of HLBI in patients with CML in the chronic and accelerated phase.

Adolescent↗
PubMed1988

[Analysis of cellular oncogenes in human osteosarcomas transplanted into nude mice].

The structures and transcripts of sixteen different cellular oncogenes (c-oncs) were studied in ten human osteosarcoma lines transplanted onto nude mice by Southern and Northern blot hybridization techniques. One osteosarcoma line (WAJI) had an amplification of c-myc gene at about ten-fold normal amount. High levels of c-myc transcripts were detected in four (MINO, OZA, SU, KiKu) of ten osteosarcoma lines. High levels of c-H-ras transcripts were also detected in four (OZA, TOMO, WAJI, SU) of ten osteosarcoma lines. Gene transcripts of c-fos were detected in only one osteosarcoma line (WAJI). Osteosarcomas containing detectable c-myc transcripts grew more rapidly in nude mice than those with undetectable c-myc transcripts. Levels of c-H-ras transcripts were apparently unrelated to the tumor growth rate.

Animals↗
PubMed1988

[Orthopedic local treatment of metastatic bone tumor].

Since 1971, we have treated 76 patients with metastatic bone tumor. Forty-eight of 76 cases were treated surgically. In 34 cases, removal of metastatic foci of the spine was attempted from the anterior and/or posterior approach. In the remaining 14 cases, resection of the tumor with or without prosthetic replacement, osteosynthesis, or amputation was performed according to the site of bone metastasis. In cases of spinal metastasis, if foci were located anteriorly, resection and replacement of vertebral body by ceramic spacer and/or bone cement was effective for both relief of pain and reduction of paralysis. If foci were located posteriorly, however, the results of simple laminectomy were poor, and those of spinal instrumentation in addition to laminectomy were satisfactory. In the treatment of metastatic foci of pelvis, scapula and long bone, the results of orthopedic surgery were satisfactory, because relief of pain and maintenance of function were achieved in almost all cases. From the standpoint of prolongation of life, 15 out of 76 cases (20%) were alive with or without disease; 13 of them were treated not only surgically but also by radiation and/or chemotherapy.

Adult↗
PubMed1987

Chromosomal gene structure of human myeloperoxidase and regulation of its expression by granulocyte colony-stimulating factor.

By using the full-length cDNA as a probe, the chromosomal gene for human myeloperoxidase was isolated from a human gene library. Comparison of the nucleotide sequence of the chromosomal gene with that of the cDNA has revealed that the human myeloperoxidase gene is composed of 12 exons and 11 introns. S1 mapping analysis of human myeloperoxidase mRNA identified the single transcription initiation site at 180 base pairs upstream of the ATG initiation codon. In the 5'-flanking region of the human myeloperoxidase gene, there are several blocks of sequences which are homologous to the sequences found on the 5'-promoter region of the human c-myc proto-oncogene. The myeloperoxidase gene is expressed in mouse myeloid leukemia WEHI-3B D+ and NFS-60 cells. When these cells were treated with human recombinant granulocyte colony-stimulating factor, the steady-state level of myeloperoxidase mRNA declined to zero within 36 h in WEHI-3B D+ cells, but not in NFS-60 cells.

Amino Acid Sequence↗
PubMed1987

The Cl- channel in hog gastric vesicles is part of the function of H,K-ATPase.

Hog gastric vesicles showed Cl- conductance when treated with Cu2+-o-phenanthroline, an S-S cross-linking reagent. An IgG monoclonal antibody caused dose-dependent inhibition of Cl- conductance that had been induced by S-S cross-linking. The antibody did not cause intervesicular aggregation, as determined by measurement of vesicle size. These results show that Cl- conductance, the stimulation and inhibition of which are regulated reversibly by S-S----2SH transformation, is due to native, physiological channels. The antibody also dose dependently inhibited the activities of H,K-ATPase and p-nitrophenyl phosphatase in gastric vesicles, but did not inhibit Na,K-ATPase obtained from dog kidney. Immunoblotting with the antibody of vesicle proteins solubilized in sodium dodecyl sulfate-polyacrylamide gel showed that the antibody binds to a 95-kDa subunit of H,K-ATPase and its dimeric 180-kDa polypeptide. The antibody-binding sites of H,K-ATPase activity and the Cl- channel for the inhibition were present on the external (cytosolic) surface of the transmembraneous ATPase. A gastric antisecretory compound, 2-methyl-8-(phenylmethoxy)imidazo[1,2 alpha] pyridine-3-acetonitrile (SCH 28080), competitively bound to the high affinity site of K+ on the internal (luminal) surface of H,K-ATPase, and its half-maximal inhibitory concentration for H,K-ATPase activity in tight vesicles was 0.2 microM in the presence of valinomycin. SCH 28080 also dose dependently inhibited opening of Cl- channels by S-S cross-linking, the regulatory site being present on the cytosolic side and more internally than the antibody binding site. The half-inhibitory concentration of SCH 28080 was 0.3 microM. The present results with the antibody and SCH 28080 indicate that the Cl- channel is part of the function of H,K-ATPase.

Adenosine Triphosphatases↗
PubMed1987

Preliminary X-ray data for a D-amino acid amino-transferase from a novel thermophilic Bacillus.

Crystals of the D-amino acid aminotransferase (D-ATA) from a novel thermophilic Bacillus species (Escherichia coli pICT113 cloned gene product) have been examined by X-ray analysis. The crystals grow as hexagonal prisms, with the symmetry of space group P61 or P65 (indistinguishable crystallographically). The cell dimensions are a = b = 135 A, c = 53 A, alpha = beta = 90 degrees, and gamma = 120 degrees. The unit cell has a volume of 850,000 A3 with six asymmetric units per unit cell. There is one dimer of molecular weight 62,000 per asymmetric unit, and the crystals diffract to 2.7 A.

Bacillus↗
PubMed1987

[A human case of pulmonary dirofilariasis suspected to be lung cancer].

A 50-year-old man in Kitakyushu City visited a hospital for evaluation of an opacity in a routine chest X-ray film. After examinations by bronchoscopy and chest roentgenography, a clear coin lesion was found in the middle lobe of the right lung and was suspected to be a lung cancer. A right thoracotomy was performed to remove it. The lesion (10.6 X 9.8 mm) was a well-defined ellipsoid granuloma due to a foreign body with a central zone of necrosis surrounded entirely by a fibrous wall. In a cross section of the dissected granuloma two degenerated worm-like structures were revealed. The sections of the worm (216-240 X 296 mu m at the greatest diameter) had at least a 3-layered cuticle and prominent internal longitudinal cuticular ridges but no external cuticular ridges. The lateral chords were as high as the muscle layer which consisted of abundant somatic muscle (more than 30 muscle cells/quadrant). Judging from these morphological characteristics, the pulmonary granulomatosis was diagnosed to be due to an immature dog heartworm, Dirofilaria immitis. This identification was further supported by results from immunological methods. The present case is the 56th human case of dirofilariasis (the 39th as a pulmonary case only) in Japan.

Diagnostic Errors↗
PubMed1987

Molecular cloning and characterization of cDNA for human myeloperoxidase.

Partial amino acid sequence of human myeloperoxidase was determined, and a 41-base oligonucleotide containing deoxyinosines at four positions was chemically synthesized. By using the oligonucleotide as a probe, cDNA clones for human myeloperoxidase were isolated from a cDNA library constructed with mRNA from human promyelocytic leukemia HL-60 cells. One of the clones containing a 2.6-kilobase insert was subjected to nucleotide sequence analysis. The sequence was found to contain an open reading frame, 2,235 nucleotides coding for a protein of 745 amino acids with a calculated Mr of 83,868. The heavy chain of myeloperoxidase, consisting of 467 amino acids, was located on the COOH terminus half of the protein. The RNA specified by the cDNA was prepared using SP6 RNA polymerase and translated in rabbit reticulocyte lysates, and the product was identified as human myeloperoxidase by immunoprecipitation with rabbit anti-human myeloperoxidase antibody. By Northern hybridization analysis of RNA from leukemic cells, it was shown that myeloperoxidase mRNA is abundantly expressed in human promyelocytic HL-60 and mouse myeloid leukemia NFS-60 cells. Furthermore, the results of Southern hybridization analysis of human genomic DNA suggest that there are one or two genes for myeloperoxidase in the human haploid genome.

Amino Acid Sequence↗
PubMed1987

Characterization of recombinant human granulocyte-colony-stimulating factor produced in mouse cells.

Mouse C127I cells were transformed with a chimeric plasmid consisting of bovine papillomavirus DNA and human granulocyte-colony-stimulating factor (G-CSF) cDNA placed under the control of the SV40 early promoter. The transformed cells secreted constitutively a high level of human G-CSF, 10-20 micrograms/ml in a low-serum medium. The secreted G-CSF has been purified to homogeneity by a two-step procedure including gel filtration and hydrophobic column chromatography. The purified recombinant G-CSF runs as a single band with an apparent Mr of 19,000 on a polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. This value corresponds to that of the native human G-CSF purified from the medium conditioned by human carcinoma CHU-2 cells. The recombinant human G-CSF was as active as native G-CSF in vitro in supporting proliferation of mouse NFS-60 cells and stimulating colony formation from human as well as mouse bone marrow cells. When the recombinant human G-CSF was subcutaneously administrated into mice, a remarkable stimulation of granulopoiesis and splenomegaly was observed.

Animals↗
PubMed1987

Human gene coding for granulocyte-colony stimulating factor is assigned to the q21-q22 region of chromosome 17.

Granulocyte-colony stimulating factor (G-CSF) is a member of colony stimulating factors which regulate the proliferation and differentiation of hematopoietic progenitor cells. A full-length cDNA clone coding human G-CSF was used as a hybridization probe to detect homologous sequence in human-mouse somatic cell hybrids, flow-sorted human chromosomes, and in situ human metaphase chromosomes. The results indicate that the gene encoding human G-CSF is on the q21-q22 region of chromosome 17, which is involved in translocation of t(15;17) (q23;21) in human acute promyelocytic leukemia.

Animals↗
PubMed1987

Studies on endometrial cells in human pregnancy by monoclonal antibodies.

By immunizing mice with endometrial cells from pregnant women, we obtained two hybridoma clones, 9D3 (IgG1) and 9E3 (IgG3), secreting monoclonal antibodies (MoAbs) that selectively stained endometrial stromal cells having prolactin in their cytoplasm, but did not stain glandular epithelium nor small lymphoid cells. From examination of tissues, cells and cell lines, it was found that 9E3 antigen was broadly distributed among various mesenchymal cells, while 9D3 antigen was distributed on a more restricted spectrum of mesenchymal cells, skin basal cells and a promonocytic cell line (U937). Neither of these antibodies stained peripheral blood cells, bone marrow cells or endometrium of the proliferating phase. Furthermore, 9D3 MoAb reacted with all melanoma cell lines and squamous cell lines examined. The antigenic molecule that was recognized by 9D3 MoAb had a molecular weight of 94,000.

Animals↗
PubMed1987

Comparative study of serum lipoproteins separated by the method of cellulose acetate membrane electrophoresis in humans and rats.

The evaluation of lipoproteins in rat serum which was separated by cellulose acetate membrane electrophoresis was studied in comparison with that in human serum. In contrast to the human lipoprotein pattern, the top of the rat lipoprotein fraction exceeded the albumin fraction towards the anode. By the analysis of ultracentrifugation and post-heparin serum lipolytic activity, the characters of lipoprotein fractions electrophoretically separated in rat serum was confirmed as similar to human serum lipoprotein.

Animals↗
PubMed1987

Necrotizing lymphadenitis. Electron microscopical and immunohistochemical study.

Seven cases of necrotizing lymphadenitis (NEL) including a pair of male siblings, a female suffering from non-bacterial meningitis, and two cases with the proliferation of monocytes and/or macrophages in the bone marrow are reported. This disease is clinically documented by the occurrence in young adults usually accompanied by painful cervical lymphadenopathy with fever and leukopenia (below 4,000/mm3). Morphological features were characterized by nuclear debris from degenerated lymphocytes and the appearance of blastoid cells and/or immunoblasts and macrophages. Ultrastructurally, tubular inclusions with a close relation to the endoplasmic reticulum were observed in various kinds of cells in the lesion. Immunohistochemical studies revealed that the ratio of helper/suppressor T-lymphocytes became low at the active stage and returned to normal range at the recovery stage. By immuno-histochemical study it was confirmed that suppressor cells mainly corresponded to large transformed lymphocytes and/or immunoblasts and helper cells were degenerated by an unknown agent. Though the pathogenesis of NEL is still uncertain, it is suggested that T-lymphocytes are mainly involved during the course of the disease and lymphocytes show cellular debris or blastoid transformation.

Adolescent↗
PubMed1987

Ultrastructural studies of the skin and cultured fibroblasts in I-cell disease.

The skin and cultured fibroblasts from a patient with I-cell disease were examined by electron microscopy. Multiple vacuolations were seen in fibroblast and/or histiocyte-like cells, secretory cells of eccrine glands, and Schwann cells of the skin. Vacuolar inclusions were single membrane-limited, and contained a few reticulo-floccular and vesicular materials, endothelial cells of the dermal capillaries contained other types of inclusions, which were also membrane-limited, more electron dense, and multivesicular. The epidermis and pilosebaceous appendages seemed to be normal. Cultured skin fibroblasts contained prominent inclusions which varied in size and morphology. Acid phosphatase activity was seen in some of those inclusions, indicating their derivation from lysosomes. These findings suggest that the I-cell disease is a type of lysosomal storage disease and that electron microscopic examination of normal-appearing skin in this disease may contribute to the diagnosis.

Cells, Cultured↗