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Biomedical subjects

A Okano

Publications and source records attributed to A Okano.

At least 91 records · Page 5Linked to original sources
PubMed1993

Changes in hematopoiesis-supporting ability of C3H10T1/2 mouse embryo fibroblasts during differentiation.

To investigate the functional change of stromal cells along with differentiation, we used a differentiation-inducible mouse embryo fibroblast cell line, C3H10T1/2 (10T1/2). Stably determined preadipocyte and myoblast cell lines were established after a brief exposure of 10T1/2 cells to 5-azacytidine. These cell lines terminally differentiated into adipocytes and myotubes, respectively, under appropriate conditions. The hematopoiesis-supporting ability of each 10T1/2-derived cell line was examined by coculture with FACS-sorted murine hematopoietic stem cells (Thy-1lo c-kit+ Lin-). The number of granulocyte-macrophage progenitors (CFU-GM) was slightly reduced after 7 days of culture with parent 10T1/2 fibroblasts, whereas a marked increase in CFU-GM number was observed when the cells were cultured on preadipocytes. Mature adipocytes and myogenically determined cell lines, on the other hand, did not support CFU-GM growth. Further, Northern analysis showed that the preadipocyte cell line acquired the ability to produce a significant amount of stem cell factor (SCF), interleukin-6 (IL-6), leukemia inhibitory factor, and macrophage colony-stimulating factor mRNAs in response to IL-1 or lipopolysaccharide stimulation. Terminal adipocytic differentiation resulted in reduced ability to express these cytokine mRNAs. Similarly, highest IL-6 activity was detected in the supernatant of preadipocyte culture, whereas adipocytes did not secrete IL-6 even after IL-1 stimulation. Interestingly, hematopoiesis-nonsupporting myoblasts and myotubes also expressed abundant SCF mRNA, suggesting that SCF, per se, may not be sufficient for stem cell growth and survival. The 10T1/2-derived cell lines could provide a valuable tool to aid in the analysis of stromal cell development and the search for novel stromal factors.

Adipose Tissue↗
PubMed1993

Effect of thiol compounds on in vitro development and intracellular glutathione content of bovine embryos.

The purpose of this investigation was to determine the effect of beta-mercaptoethanol (beta-ME) and cysteamine, low-molecular-weight thiol compounds, on the development and intracellular glutathione content of bovine embryos obtained by in vitro fertilization of in vitro-matured oocytes. Embryos developed to the 6-8-cell stage after in vitro fertilization were cultured without feeder cells in TCM-199 containing 10% fetal calf serum with or without beta-ME or cysteamine. The percentage of embryos that developed to the blastocyst and hatched blastocyst stages were significantly higher in medium containing beta-ME or cysteamine. Also, total intracellular glutathione levels were higher for embryos cultured in the medium with beta-ME or cysteamine than for those cultured in medium without thiol compounds. Moreover, when buthionine sulfoximine, a specific inhibitor of glutathione synthesis, was added to medium containing thiol compounds, there was a reduction both in the development of embryos to the blastocyst stage and in intracellular glutathione content. These results indicate that the inclusion of low-molecular-weight thiol compounds aids the in vitro development of bovine embryos without feeder cells and that the effect of thiol compounds is mediated through the increase of intracellular glutathione levels.

Animals↗
PubMed1993

The production of granulocyte colony-stimulating factor and interleukin 6 by human bone marrow stromal cells in aplastic anemia.

Stromal cells are known to produce granulocyte colony-stimulating factor (G-CSF) and interleukin 6 (IL-6). To assess the function of bone marrow stromal cells in the pathological state, we studied their in vitro ability to produce G-CSF and IL-6 in 9 patients with aplastic anemia and 5 normal volunteers, using an enzyme immunoassay method. Constitutive production of IL-6, but not G-CSF, was detected in almost all cases. Interestingly, the inducible production of G-CSF and IL-6 by stromal cells after stimulation with interleukin-1 and/or lipopolysaccharides was severely depressed in 3 patients with aplastic anemia. Our results may reflect poor hematopoietic activity in the bone marrow in some cases of aplastic anemia.

Adolescent↗
PubMed1993

[A case of T-cell lymphoma showing marked bronchial invasion].

We report a case of malignant lymphoma with bronchial involvement in a 35-year-old male patient. On admission, chest X-ray revealed enlargement of bilateral hilar lymph nodes and expansion of the superior mediastinal space. CT and MR images showed that the tumor shadow appeared to wrap around the left and right main bronchi. Bronchoscopy revealed that the entire periphery of the left main bronchus was constricted. Transbronchial biopsy specimens, which revealed findings consistent with non-Hodgkin's lymphoma, were positive for surface markers of CD2, CD4, and CD8 and negative for those of CD19 and CD20. After MACOP-B treatment, the patient entered complete remission. He received 41.4 Gy radiotherapy thereafter, and has shown no signs of recurrence during continuing follow-up.

Adult↗
PubMed1993

Thrombopoietic effects of interleukin-6 in long-term administration in mice.

To further investigate the thrombopoietic and adverse effects of interleukin-6 (IL-6), 2 or 10 micrograms/day of recombinant human (rh) IL-6 was administered intraperitoneally (i.p.) to mice for up to 30 days. IL-6 increased platelet count, which plateaued at a level 30 to 40% higher than control after 5 days of treatment. This cytokine also maintained the high platelet count for the duration of treatment. The count exceeded normal levels 7 days after cessation of the 30-day treatment. IL-6 also induced a remarkable increase in the size but not the frequency of megakaryocytes in bone marrow sections. The number of bone marrow colony-forming units megakaryocyte (CFU-MK) and colony-forming units granulocyte-macrophage (CFU-GM) was not augmented by the administration of IL-6 in this protocol, while spleen progenitors were significantly stimulated. Small but significant increases did occur in the number of bone marrow megakaryocytes and CFU-MK, and in the proportion of CFU-MK in the DNA synthetic phase in mice treated with 10 micrograms/day of IL-6 for 30 days. Electron microscopic examination of bone marrow demonstrated that IL-6 remarkably developed the distribution of the demarcation membrane system (DMS) in mice treated for 30 days, with little change in mice treated for 5 days. The administration of 2 micrograms/day for 30 days induced a 2.2-fold increase in fibrinogen. No changes were observed in the hepatic or renal functions. Histologic and immunofluorescence studies on the kidneys revealed no significant changes compared with controls, indicating that proliferation of the glomerular mesangium did not occur. No neutralizing antibodies were detected in mice treated for 30 days. We conclude that the long-term administration of IL-6 in mice stimulates megakaryocyte maturation and platelet production with few adverse effects, and that this cytokine may be a candidate for the treatment of thrombocytopenia in humans.

Animals↗
PubMed1993

[A case of familial antithrombin III variant complicated by recurrent pulmonary infarction].

We present a 36-year-old male who developed pulmonary infarction secondary to antithrombin III variant. He was admitted to our hospital with progressive chest pain and hemosputum. The presence of multiple defects on 99mTc perfusion scan indicated the diagnosis of pulmonary infarction. He had a past history of pulmonary infarction 4 years before this episode as well as a family history of recurrent thromboembolic disease. Through coagulation studies revealed that the concentration of antithrombin III antigen and progressive antithrombin activity were within normal limits, while heparin cofactor activity was decreased markedly. Of 5 people in his family, 4 proved to have low heparin cofactor activity. These data suggest that functional abnormality of antithrombin III seems to be closely associated with pulmonary infarction.

Adult↗
PubMed1992

Acquisition of interleukin-3 independence in FDC-P2 cells after transfection with the activated c-H-ras gene using a bovine papillomavirus-based plasmid vector.

Since the ras family of proto-oncogenes is supposed to be involved in leukemogenesis by point-mutational activation, we studied the effect of the activated ras gene on the growth of a murine interleukin-3 (IL-3)-dependent cell line, FDC-P2. The human activated c-H-ras gene was transfected into FDC-P2 cells by electroporation using a high-level expression vector, BMGhph, which contains a partial DNA sequence from bovine papillomavirus (BPV) and a hygromycin B (hmB)-resistant gene as a selectable marker. The transformed FDC-P2 cells showed a high incidence of IL-3-independent growth and tumorigenicity in nude mice. These clones did not express or secrete IL-3, suggesting the acquisition of IL-3 independence by a nonautocrine mechanism. The high incidence of autonomous growth may be due to the use of the BMG vector, because (1) the activated ras gene in pBR322 vector (pHs-49) was not so efficient in the induction of IL-3 independence, (2) the c-H-ras genome copies per cell increased in number up to about 50 copies by using the BMG vector, and (3) cotransfection with the activated ras gene and the BPV gene in separate plasmids partly enhanced the incidence of autonomous growth without increasing the copy number of the ras gene compared with transfection with the activated ras gene alone. The present study supports the idea that the activation of ras gene is an important step in malignant transformation of hematopoietic cells and suggests that the BPV gene products may cooperate with ras gene activation probably by affecting the cellular genes that may be involved in multistep leukemogenesis. The BMG vector may be useful to test the transforming ability of oncogenes whose oncogenic potential is relatively low.

Animals↗
PubMed1992

A difference in the proteins found in young adults of inbred strains of Drosophila melanogaster which correlates with genetically-determined, long or short life span.

One-dimensional electrophoresis was performed on extracts of flies collected from across all ages. Protein gel patterns were compared for two strains of Drosophila melanogaster with distinctly long and short adult life spans that result from different alleles of longevity genes. An inter-strain difference was observed in the changes in protein pattern in the 77 kDa region in period of day 0-5 after emerging. We propose that the protein involved is a product of autosomal longevity alleles A1 and A2 at the Jm A locus and is related to development of longevity potentials in the preimaginal stage.

Animals↗
PubMed1992

Purification and characterization of a protein associated with genetically-determined longevity difference in Drosophila melanogaster.

Earlier studies have shown a correlation between the presence of a 77 kDa protein in the proteins extracted from young adult Drosophila melanogaster (D.m.) and the autosomal longevity allele. A2 at the JmA locus. In this study, a 77 kDa protein has been isolated from pupae of D.m. of a long-lived strain of genotype A2A2, and was purified by DEAE chromatography, ConA column chromatography, and two cycles of gel filtration. The purified protein has a molecular weight of 76,600 (by SDS-PAGE), an isoelectric point of pH 6.5, and molar extinction coefficient A(280(1%) = 18.3. It is a glycoprotein containing 3.3% hexose. Supplementing the food of D.m. with the purified protein at 5 x 10(-4) micrograms/ml, beginning at day 5 after emergence, resulted in an increase in the survival rate and maximal life span of both short-lived and long-lived strains of D.m.

Animals↗
PubMed1992

[Etiology of pneumonia and host defense].

To investigate the relationship of efficacy of chemotherapy to host defense, we reviewed the clinical features, treatment and outcome in 648 patients with acute pneumonia (424 males and 224 females; mean age, 65 years) treated between 1984 and 1989. Pneumonia complicated pulmonary disease in 336 patients (52%) and complicated systemic disease in 258 (40%). Pneumonia pathogens were diagnosed in 346 patients (53%); the five major pathogens were S. pneumoniae (19%), H. influenzae (16%), S. aureus (14%), K. pneumoniae (14%) and P. aeruginosa (11%). The incidence of K. pneumoniae infection were decreased from 18% to 3.5% and that of S. aureus increased from 10.9% to 26.3% during the study period. The incidence of S. aureus and of P. aeruginosa infection was much higher in patients with nosocomial pneumonia, systemic disease, or serum protein concentration under 6.5 g/dl. beta-lactamase antibiotics were administered to 70% of all patients, with an efficacy rate of 74.9% of after the first course of antibiotics therapy. The efficacy rate was decreased in patients with nosocomial pneumonia, systemic or pulmonary disease, or malnutrition. The data presented in this study indicate that the risk of pneumonia must be taken into careful consideration in patients with compromising complications.

Acute Disease↗
PubMed1992

[A case of empyema with subphrenic abscess].

A 75-year-old female was admitted to our hospital with complaints of fever, cough and left hypochondralgia. She had been operated for cholecystectomy ten years ago. Chest roentgenogram indicated bilateral pleural effusion. Tube drainage was done to the left thorax and empyema was caused by Bacteroides fragilis and Escherichia coli (E. coli). Though antibiotic therapy was already being conducted, the left hypochondralgia persisted. A CT scan and MRI demonstrated local subphrenic abscess around the spleen due to E. coli. Tube drainage was conducted to the subphrenic abscess under ultrasound control and and the symptoms disappeared rapidly. The present results show that examination of the abdomen is necessary for empyema with complication of compromised host. The past history of abdominal surgery and disturbance in the biliary tract should also be considered.

Aged↗
PubMed1992

Optical resolution of (+/-)-all-trans-3-hydroxyretinal by use of high-performance liquid chromatography.

Racemic all-trans-3-hydroxyretinal (3-OH-RAL) (1) was converted by a reaction with (-)-camphanic acid chloride (CpCl) into a diastereomixture of camphanates (2a) and (2b) which was separated by preparative high-performance liquid chromatography (HPLC) to give two esters (2a) and (2b) in pure state, respectively. Saponification of (2a) and (2b) independently afforded optically active (3S)- and (3R)-3-OH-RALs (3a) and (3b), respectively, whose absolute structures were determined by circular dichroism (CD) spectra. Racemic 3-OH-RAL was separated to two peaks by HPLC using chiral column (ChiraSpher, Merck). Cochromatography with authentic specimens (3a) and (3b) showed that the peak with a short retention time corresponded to (3R)-isomer and the other to (3S).

Chromatography, High Pressure Liquid↗
PubMed1992

[Case of adult primary varicella pneumonia].

We report a case of adult primary varicella pneumonia. A 34-year-old man was admitted to our hospital with fever, dry cough and eruptions. He had no history of chicken pox and his sons had contracted varicella 2 weeks before the onset of his symptoms. Chest X-ray showed diffuse nodular shadows in both lungs. The diagnosis of primary varicella pneumonia was made based on family history, typical eruptions and high titer of antibody against Varicella zoster virus. An electron micrograph indicated this case to be primary varicella pneumonia with fibrosis and edema of interstitial spaces and the presence of virus-like particles in cells. The patient was treated with antibiotics, an antiviral agent and immunoglobulin. The clinical symptoms and diffuse nodular shadows resolved with this treatment.

Acyclovir↗
PubMed1992

[A case of secondary lung cancer of cystadenocarcinoma showing multiple thin-walled cavities].

A 74-year-old woman was admitted to our hospital with the complaint of general fatigue. Chest X-ray revealed multiple thin-walled cavities in both lung fields. Cystadenocarcinoma in the liver and secondary lung cancer were diagnosed on the basis of chest X-ray, abdominal CT, and lung biopsy. She died of respiratory failure, in spite of treatment with chemotherapy for 8 months. At autopsy, the liver mass was diagnosed as cystadenocarcinoma, which had developed from a benign cystadenoma with the capacity to produce mucinous material. Thin-walled cavities in the lungs showed the same histological findings. Therefore, the mechanism of cavity formation in this case was speculated to have been metastasis of mucin-producing cystadenocarcinoma, with entry of air secondary to communication with the terminal bronchiole.

Aged↗
PubMed1992

[A case of bronchiolitis obliterans organizing pneumonia with positive anti Jo-1 antibody preceding polymyositis].

We describe a 58-year-old female with BOOP associated with polymyositis. Four months prior to the appearance of distinctive manifestation of polymyositis, she presented with a two-week history of cough, dyspnea on exertion, and fever. Chest roentgenogram demonstrated bilateral basal infiltrative shadows. The patient was treated with prednisolone, 30 mg/day because of progressive hypoxemia. Open lung biopsy revealed organizing masses of granulation tissue extending from the respiratory bronchioles into the intra-alveolar spaces, which was consistent with BOOP. She developed muscle pain in her legs, fever, dry cough, hypoxemia, and high CPK value in the course of tapering of steroid dose. The findings of biopsy from the left rectus femoris muscle were compatible with polymyositis. Retrospective study of the patient's serum on admission showed positive anti Jo-1 antibody.

Antibodies, Antinuclear↗
PubMed1991

A new method of gene transfer into hematopoietic progenitors using liquid culture with interleukin-3 and interleukin-6.

The technique of gene transfer into hematopoietic cells is expected to offer a new form of therapeutics. As a result of studies on a gene-delivery system using granulocyte-macrophage colony-forming units (CFU-GM), a type of hematopoietic progenitor, we have established a technique for efficient gene transfer into CFU-GM. DGL, a retrovirus vector containing the SV40 promoter and the neomycin resistance gene, was constructed and found to transfer genes effectively into murine CFU-GM, which subsequently expressed the neomycin resistance gene. After gene transfer of murine non-adherent bone marrow cells precultured in liquid culture with recombinant murine IL-3 (rmIL-3) and recombinant human IL-6 (rhIL-6) for 6 days, gene transferred CFU-GM in bone marrow cells were able to proliferate 5-10-fold and the ratio of gene transferred CFU-GM to total CFU-GM reached 70-100% from less than 1% in the liquid culture with rmIL-3, rhIL-6 and neomycin for 6 days. Using this protocol, we have been able to obtain large amounts of highly concentrated gene-transferred CFU-GM for fundamental research on CFU-GM gene-delivery systems.

Animals↗