[Pulmonary tuberculosis with unusual x-ray findings (author's transl)].
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Biomedical subjects
Publications and source records attributed to T Kimoto.
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The presence of C3 receptor sites on the cell surfaces of WI-38 fibroblasts was reported in a previous paper. Here the effect of dexamethasone sodium sulfate of C3 receptor site function was studied. The incubation of WI-38 fibroblasts with dexamethasone sodium sulfate produces the biphasic mode of action, i.e., the growth-stimulating phase with low doses (90--230 mug/ml) and the growth-inhibiting phase with high doses (450--900 mug/ml). The function of C3 receptor sites on WI-38 fibroblasts seems to be very stable and cannot be suppressed by the pretreatment of WI-38 fibroblasts with dexamethasone in high concentrations, where the cell growth is inhibited.
The peripheral blood of an acute myelomonocytic leukemia patient has been cultured for 16 months. The culture is at present at the 140th population doupling level. The cultured cells have the characteristics of so-called lymphoblastoid cells and proliferate actively as individual cells in small clusters, or in large clumps consisting of large mononuclear cells. Some of these cells appeare to be lymphoid, but the majority are immature mononuclear cells with a tendency to lobulate. They gave a weakly positive peroxidase reaction at the beginning of cultivation, and have given a strongly positive esterase reaction persistently. The cytoplasm shows ciliary or tail-like projections as the cell matures. Complement (C3) receptor and IgG receptors are found on the cell surface, and active phagocytosis is mannifest. Colloidal iron particles or viable red blood cells attached to the cell membrane suggesting possible differentiation to reticulum cells or macrophages. The cultured cells are mostly diploid but some cells show chromosome abnormality. Herpes type virus was foun in the nucleus, cytoplasma and on the cell membrane. The transplanatation of cultured cells to the cheek pouch of hamsters produced small tumors with histological findings resembling reticulum cell sarcoma.
The presence of C3 receptor sites on WI-38 human diploid fibroblasts was reported in the previous paper. In this report we wish to describe that WI-38 fibroblasts carry C3b and C3d receptor sites on the cell surfaces.
Formation of prominent multinucleated giant cells (MGC) was observed in monolayers of a clonal line of BHK-21 cells (BHK-21-528) when infected with Japanese encephalitis virus (JEV). MGC were first observed 3 to 4 days after infection and cytopathic changes proceeded thereafter. Formation of MGC is a typical cytopathic change in this clonal cell line. Virus titer in 50% tissue culture infective dose (TCID50) equaled that in 50% MGC-forming dose. Virus titer in TCID50 was approximate to plaque-forming units (PFU) in the same host cells. An ability of JEV to form MGC was maintained at least for six serial passages in BHK-21-528. It was inactivated by heating at 56 C for 3 min. All JEV strains, except an attenuated live vaccine strain, induced formation of MGC in BHK-21-528 cells. Red blood cells of several animal species were not adsorbed to MGC induced by JEV. The MGC-forming ability of JEV was specifically neutralized by anti-JEV serum. By fluorescence antibody technique, the MGC were specifically stained by anti-JEV antibody conjugated with fluorescein isothiocyanate. Immunization of animals with lysates of the MGC resulted in production of antibodies against JEV, but no antibody against other viruses which have been reported to induce MGC formation. From these evidences, it was concluded that JEV induced formation of MGC in BHK-21-528.
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A cell line (HGC-27) was established by culture of the metastatic lymph node from a gastric cancer patient diagnosed histologically as undifferentiated carcinoma. HGC-27 cells were polygonal or short spindle-shaped and adhered to glass surfaces as a monolayer. The cells were probably derived from gastric cancer cells, as their origin from mesenchymal tissues can be excluded morphologically and enzyme-histochemically. Enzyme activities were generally negative or low, except for adenosine triphosphatase, lactic dehydrogenase and leucine aminopeptidase. These scanty findings might reflect the undifferentiated character of the original tumor cells. The cloning efficiency was 5.3% in liquid medium and 1.0% in soft agar. The doubling time was about 17 hr. Chromosomal analysis revealed a mode of 109 and 110 chromosomes.
A permanent cell line (HLC-1) was established from the pleural effusion of a human lung adenocarcinoma. The cell line was characterized by the monolayered and multilayered organoid growth of epithelioid cells with the doubleing time of about 33 hr and the modal chromosome number of 68. Cloning efficiency was 17.9% in liquid medium and 8.3% in soft agar. The cell produced a large amount of epithelial mucin. Electron microscopic examination revealed many secretory granules and terminal bars. They formed spherical aggregates in a gyratory culture which showed adenocarcinoma-like tubular structures histologically. Enzyme-histolochemically, they showed the characteristics of lung adenocarcinoma cells except for a few enzymes such as glucose-6-phosphatase and ATPase. Heterotransplantation of the cells produced the tumor. These characteristics confirm that HLC-1 cell line is a human lung adenocarcinoma cell line.
Adult mongrel dogs, weighing 10-17 kg. were anesthetized with Nembutal and cervical and lumbosacral laminectomy was performed. The spinal subarachnoid space was blocked by extradural ligation at the level of the C4 to interrupt CSF communication between the cranial and spinal space. Polyethylene catheters were placed in the cervical and lumbosacral subarachnoid space, and artificial Mock CSF buffer, pH 7.35-7.40, containing inulin of 25 mg/dl or 14C-inulin of 1.5-2 muCi/dl as a tracer was perfused in the sacro-cervical direction through the catheter. After a steady state of perfusion was acommplished, the CSF was collected from the outlet catheter. Production and absorption rate of the CSF were calculated after Pappenheimer and Heisey's equation. 1) Effects of CSF pressure on the rate of production (Vf) and absorption (Va) of CSF and on the difference between outflow fluid rate (Vo) and inflow fluid rate (Vi) were studied within the pressure range of -100 to +600 mmH2O. Then, regression lines were calculated by means of the least square method. See Article. Vf was little affected by changes in CSF pressure, while Va increased linearly as CSF pressure elevated. This suggests that the spinal subarachnoid space plays an important role as a site of CSF absorption when the intracranial pressure increases. Vo-Vi, that is difference between absorption and production rate, decreased linearly as the CSF pressure increased. 2) Under a constant CSF pressure of +200 mmH2O, the effects of glucocorticoids (dexamethasone, 0.25 mg/kg and hydrocortisone, 4.15 mg/kg) and a carbonic anhydrase inhibitor (acetazolamide, 10 mg/kg) upon the production and absorption rate of CSF were determined 1/2, 1, 2 and 3 hours after intravenous administration. a) Effect of dexamethasone: The rate of CSF production was reduced to 60.5 +/- 2.4% (p less than 0.001) of the control level. The absorption rate of CSF also decreased to 59.2 +/- 6.09% (p less than 0.001) of the control. b) Effect of hydrocortisone: The production rate of CSF decreased to 67.4 +/- 6.61% (p less than 0.001), and the absorption rate to 76.5 +/- 3.94% (p less than 0.001) of the control level. c) Effect of acetazolamide: The production and absorption rate also decreased to 57.2 +/- 5.61% (p less than 0.001) and to 56.9 +/- 7.02% (p less than 0.001), respectively. 3) Pentration of tritiated dexamethasone and tritiated hydrocortisone from plasma to CSF. The penetration of tritiated dexamethasone and tritiated hydrocortisone from plasma to CSF in the spinal and cranial subarachnoid space was observed after the intravenous administration. The CSF/Plasma ratio of dexamethasone was 30.9% at 15 minutes and gradually increased to 91.5% and 93.5%, respectively, in the cranial and spinal CSF at 3 hours after the injection.
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Sheep red cells, sensitized with 19S fraction of antiserum and subsequently treated with mouse serum as the source of complement (EAC), interact with human diploid fibroblasts (WI-38 cells) and form "Rosettes". Under a scanning electron microscope, EAC have not attached directly to the cell surface of fibroblasts, but to the fine processes or microvilli of the latter, as if there were fine bridges between EAC and the surface of fibroblasts. On the other hand, the attachment of sheep red cells washed in PBS (E) or sensitized with 19S fraction of antiserum (EA) to WI-38 cells was not observed. The pretreatment of WI-38 cells with mouse serum did not inhibit the interaction of WI-38 cells and EAC. No phagocytosis of EAC by WI-38 cells was observed in the 2 hrs incubation of both cells. From these results it is suspected that the interaction of WI-38 cells and EAC is immune adherence, and that WI-38 cells have the receptor site for complement, especially for C3, on the surface of cell membrane.
In our attempts at establishing a cancer cell line from various ascites of cancer bearing patients, a cell line was successfully established from the ascites of a 63-year-old female with primary ovarian tumor (embryonal carcinoma). Histological findings of the peritoneum, due to metastasis, appeared to be cystadenocarcinoma, revealing the differentiation to non-epithelial cells which formed coarse networks and fibers, and morphologic changes of tissue cultures also reflected such histologic findings. At present the subculture has reached the 95th population doubling level, and cultured cells have assumed the morphology of mesothelial cells or fibroblasts with about 50 chromosomes. As a human malignant cell line, it is useful for the study of human malignant tumor cell.
On the basis of the previous study, on the cell interaction between malignant tumor cells and other cells, especially with lymphocytes, the present study was carried out by investigating cell to cell interaction of human malignant tumor cells and human lymphoblastoid cells such as T-cell (MOLT-4 cell) and B-cell (Burkitt lymphoma cell). As a result it has been revealed that live lymphoblastoid cells were not adhered on the cell surface of the tumor cells, nor is it ingested by tumor cells, but in thepresence of HVJ (Sendai virus: 2,000 H.A. units) it adheres slightly on the cell surface of tumor cell but no cell fusion of tumor cells and lymphoblastoid cells is observable. On the other hand, the tumor cell as well as T-cell and B-cell all have receptors to concanavalin A (Con. A) on their cell surfaces, and they show a marked cell binding such as tumor cell and T-cell, tumor cell and B-cell, and there can be observed a marked phagocytosis of lymphoblastoid cells by tumor cells. Moreover, the tumor cells that have phagocytized lymphoblastoid cells undergo a marked cell destruction within 4 hours of cell-binding and phagocytosis, which is especially prominent in the case of phagocytosis of E.B cell by tumor cell.
In reviewing 38 cases of biliary atresia, 8 cases were detected to have osseous abnormalities. We found definite generalized demineralization of bones in 3 cases, metaphyseal abnormalities in 5 cases, and fractures in 4 cases. Marked demineralization of bones was seen in cases in which the clinical course was more than 1 year. No good correlation was observed between osseous changes and severity of liver dysfunction.
Various comparative studies were made on the capacities of a cell-associated subacute sclerosing panencephalitis agent (SSPE agent, Niigata/1 strain) and a usual measles virus (Toyoshima strain) to produce measles virus hemagglutinin (MV-HAnim) in Vero cells. The results obtained by various methods, such as the hemagglutination test (HA), hemadsorption test (HAd), antibody blocking test, antibody inducing test and fluorescent antibody technique (FAT) using monospecific antibody to MV-HAnin, consistently suggested that the SSPE agent was an incomplete measles-like virus having little if any MV-HAnin, but sharing immunologically closely related viral components other than MV-HAnin with those of the usual measles virus. The significance of these findings is discussed with respects to the latency of the SSPE agent.
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