[Fibrolipoma (lipoma) of the intestine].
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Biomedical subjects
Publications and source records attributed to K Kawamoto.
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The history of brain tumor cultures is old. Established cell lines of brain tumors have increased in recent years, but reports concerning the results of these cultures are few. The results of cultures of gliomas (103 cases of astrocytoma, 9 of ependymoma, 2 of oligodendroglioma and others) obtained from 117 patients treated at our department between 1979 and 1993 were evaluated. The morphological characteristic of these glioma cells were studied, and their cellular kinetics were investigated by flow cytometry. Except for 5 cases, all tumors grew on the flask in the primary cultures, indicating that gliomas were readily cultured. Differences in morphological characteristics and cellular kinetics were not observed in cultures which had been maintained for more than 6 months in astrocytomas of grades II, III, and IV. Cells lines were successively established in 3 cases of glioblastoma and 2 of ependymoma. The results of FCM revealed that those cells which grew well on cultures had high proliferative indices.
The means of examination of small-amount samples by transmission electron microscopy (TEM) is described. In this study, the amount of cells necessary for TEM was examined by using a KMU-100 cultured cell line. Floating cells diluted with phosphate buffer solution (PBS) to several concentrations, were gathered in the tip of Beem capsules by means of centrifuge. After fixation, dehydration, and embedding with Epon 812, the samples were cut with a diamond knife, followed by electron staining, and examined by TEM. As a result, we learned that an amount of cells over 1 x 10(3) could be observed by TEM. In spite of cell damage through this procedure, the microscopic structures of the cells were relatively maintained compared with the original cell monolayers. In the near future, it may be possible to examine the sorted cells by flow cytometry (FCM) with TEM.
Four patients with human T-lymphotropic virus type I associated myelopathy (HAM) were examined ophthalmologically, and serum titers to human T-lymphotropic virus type I (HTLV-I) in 11 patients with nonspecific retinal vasculitis or uveitis were determined. All of the patients lived in the Izumo area. All 4 patients with HAM (Cases 1 to 4) had vitreous opacities. Of the 11 patients with nonspecific retinal vasculitis or uveitis, 3 (Cases 5 to 7) had elevated titers to HTLV-I. We believe that HTLV-I infection may be involved in the causes of retinal vasculitis and uveitis in patients living not only in the endemic area but also outside the endemic area.
In this study, we describe the biological characteristics and cellular kinetics (analyzed by flow cytometry) of a human glioblastoma cell line (KMU100) which had been successively maintained for 10 years. The observation GFAP (glial fibrillary acidic protein)-positive cultured cells were still noted after 275 successive passages in KMU100. GFAP stains, even though weaker, was still observed beyond the 275th passage. The doubling time was found to be 15 hours at the 490th passage. The modal chromosome number was 71, and showed a high triploid pattern. Analysis of DNA histogram by flow cytometry revealed that the cellular proliferative activity was higher in the cells belonging to the cultured cell line than in the original tumor cells from which the cell line was derived.
The significance of phospholipase D in metabolic regulation is being recognized. The products of phospholipase D action are phosphatidic acid and lysophosphatidic acid (LPA). Whether these lipid products exert their effect through plasma membrane receptor-mediated mechanism is not yet clear. In the present study we have shown the presence of specific receptors for LPA in plasma membranes of PC12 pheochromocytoma cells. LPA evoked a transient rise in [Ca2+]i concomitant with formation of IP3. LPA also stimulated dopamine release from PC12 cells. These results indicate that LPA stimulates catecholamine release via specific LPA receptors presumably by activating the IP3 mediated Ca2+ release.
We demonstrated the enhancement of HLA-DR antigen expression on cultured laryngeal carcinoma cells (Hep 2) by in vitro cultivation with LAK cells using flow cytometric and immunohistological analysis. For in vitro cultivation of tumor cells with LAK cells, we used newly developed experimental systems (the Transwell double-dish system and experimental three-dimensional tumors). In flow cytometric analysis, expression of HLA-DR antigen on tumor cells was compared before and after co-cultivation with LAK cells. When tumor cells were cultured separately with LAK cells in a Transwell Petri dish and the expression of HLA-DR antigen on tumor cells was analyzed by flow cytometry, the expression of HLA-DR antigen on tumor cells was increased in a dose-dependent manner related to the number of LAK cells used. Furthermore, when anti-interferon-gamma monoclonal antibody was added to the experimental system, enhancement of HLA-DR antigen expression was blocked. These findings were consistent with immunohistological studies, in which experimental three-dimensional Hep 2 cell tumors were established in double-layered agar with/without being co-cultivated with LAK cells. The expression of HLA-DR antigen in this system was significantly increased when compared to such expression before cultivation with LAK cells. These findings suggested that the culture systems employed in this study could be a possible model for examining solid tumor in vivo biological responses. This enhanced expression of HLA-DR antigen may also represent one of the multi-factorial responses seen with adoptive LAK cell immunotherapy for solid tumors.
HPC-1 antigen is a neuron-specific 34 kDa protein, identical to p35A (syntaxin), and is thought to play important roles in docking or fusion of synaptic vesicles to presynaptic active zones. In the present study we analyze the distribution of HPC-1 antigen in rat cerebellum by a cryoimmunogold technique using an antibody against the fusion protein of beta-galactosidase and the HPC-1 antigen. HPC-1 antigen was detected at high density on the plasma membranes and synaptic vesicles of presynaptic boutons which formed synapses with dendrites of Purkinje cells, and on the plasma membranes of parallel fibres in the cerebellar molecular layer. In the granule cell layer, gold particles were also detected on the endoplasmic reticulum, nuclear membranes and the plasma membranes of granule cells. Presynaptic membranes and synaptic vesicles in glomeruli were also labelled by gold particles. To determine the topology of HPC-1 antigen on the membranes, the synaptosome fraction prepared from rat cerebellum was embedded in agarose, and processed for the pre-embedding protein A-gold technique. Intact synaptosomes were not labelled by gold particles. However, when fixed in hypotonic fixative to rupture plasma membranes, or when ruptured after fixation in normotonic fixative, the cytoplasmic surfaces of presynaptic membranes and synaptic vesicles were labelled by gold particles. These results suggest that most of the epitopes of HPC-1 antigen are located on the cytoplasmic surface of plasma membranes and synaptic vesicle membranes.
The nerve growth factor (NGF) is widely distributed in the target tissues of sympathetic neurons. Hemopoietic organs such as the bone marrow (BM) and spleens are known to be innervated by noradrenergic sympathetic neurons. Some of their constitutive cells express NGF receptors (lymphocytes and stroma cells) and its biologic effects have been extensively studied in the immune system and inflammatory responses. However, the effects of NGF on hemopoiesis have been little examined. Recently, we demonstrated that NGF promoted mast cell colony formation from murine BM cells (BMC) or BMC-derived cultured mast cells and induced the phenotypic changes in standard hemopoietic assays. Besides, we demonstrated NGF-enhanced murine neutrophil survival and functional properties. In this study, in order to investigate NGF activities on neutrophil differentiation, we examined granulocyte-macrophage (GM) colony formation from murine BMC or spleen cells (SC) in the methylcellulose culture. We also assessed mast cell colony formation. GM colonies were counted on day 5 and mast cell colonies were counted on day 20 in culture. Although NGF alone (50 ng/ml) neither supported GM nor mast cell colony formation, addition of various doses of NGF to the suboptimal dose of pokeweed mitogen-stimulated SC-conditioned medium (2.5%) or interleukin 3 (50 U/ml), well-known colony-stimulating factors, increased the number of GM and mast cell colonies from both BMC and SC in a dose-dependent manner. These colony formation-enhancing effects of NGF were inhibited by the addition of neutralizing sheep anti-NGF antibodies. The results suggest that NGF may act to develop granulopoiesis including neutrophil and mast cell differentiation in cooperation with hemopoietic factor(s) during inflammatory processes.
OBJECTIVE: Despite advances in gastrointestinal radiologic techniques, findings from barium enema studies cannot be used reliably to predict the histologic nature of a colonic submucosal tumor. Therefore, we generated a new and comprehensive classification system for all colonic submucosal tumors based on a correlation between their radiologic characteristics and their histopathologic features. MATERIALS AND METHODS: Between 1978 and 1990, 89 patients had 95 colonic submucosal tumors diagnosed on the basis of barium enema studies and confirmed by biopsy or surgery. Of these, 39 patients (44%) were symptomatic and 50 patients (56%) were asymptomatic. Single lesions (82 cases) and multiple lesions (12 cases of lymphoma and one case of blue rubber bleb nevus) were observed. The 95 lesions included 21 carcinoids, 27 malignant lymphomas, 17 lipomas, 10 lymphangiomas, six leiomyosarcomas, five leiomyomas, and nine others. We analyzed the radiologic findings and classified these lesions according to their appearance. RESULTS: All tumors were classified on the basis of their morphologic appearance into five types: (1) wide-based sessile lesion with gradually sloping margin and smooth surface (17 cases); (2) wide-based sessile lesion, more polypoid than the first type, with a smooth surface, with or without a central depression (49 cases); (3) wide-based sessile lesion with lobulated surface (six cases); (4) pedunculated lesion with smooth or granular surface (17 cases); (5) unclassified, not any of the previously mentioned types, may be diffusely stenotic or aneurysmal (six cases). The most common finding was the wide-based sessile lesion with a smooth surface and a possible central depression; next most common was the wide-based sessile lesion with gradually sloping margin and smooth surface; next was the pedunculated lesion with a smooth or granular surface. Eighty-six percent of the lesions were single; 14% were multiple. Histologic confirmation revealed mainly lymphoma (29%) and carcinoid (22%), of which the most common finding was a wide-based sessile lesion with a smooth surface, with or without a central depression. CONCLUSION: This new classification based on radiologic characteristics covers all varieties of colonic submucosal tumors. It can serve as a guideline for predicting a specific diagnosis of a colonic submucosal tumor on the basis of radiologic findings.
The antiproliferative effect of lymphokine activated killer (LAK) cells on head and neck tumor cells has not previously been elucidated. We studied the inhibitory effects of recombinant interleukin-2 activated lymphocytes on tumor colony formation in semisolid agar, using head and neck tumor cells prepared from established tumor cell lines (K562, HT29, HLaC78) and xenografted head and neck squamous cell carcinomas on nude mice (XKN, XLL, XFL, XKF). LAK cells demonstrated a significant inhibitory effect on colony formation. The effects of LAK cells on cultured tumor cell lines were evaluated in a dose dependent manner using effector: target ratios. In addition, the colony formation of tumor cells derived from xenografted nude mouse was inhibited by LAK cells. These results suggest that LAK cells generate an antiproliferative effect on head and neck tumor cells.
A KB cell line resistant to cisplatin (KB-rc cell) was successfully established by exposing KB cells to a gradually increasing dose of cisplatin in vitro. The concentration of cisplatin required for 50% inhibition of KB-rc cell proliferation was 1.0 micrograms/ml, while that of KB cells was 0.5 micrograms/ml. Studies of KB-rc cell kinetics after treatment with cisplatin, using flow cytometry, showed that the G2M phase block was inhibited on day 3, and that G0G1 phase cells started to increase on day 5. Therefore, cisplatin-resistance is related to inhibition of the G2M phase block. The decrease of KB-rc cell viability with cisplatin was accelerated by the addition of a Ca antagonist (verapamil) and S phase cells increased on day 3. Verapamil may therefore be useful for enhancement of the effect generated by cisplatin on cisplatin-resistant tumor cells.
As a basic study, an analysis was made, using flow cytometry (FCM), of a mixture of KB cells and lymphocytes after Propidium Iodide (PI)- Fluorescein Isothiocyanate (FITC) double strain. As a result, DNA histogram of only KB cells could be drawn. As a clinical application an analysis was made, using FCM, of tumor of the head and neck after double stain. It enabled the drawing of DNA histogram of only epithelial cells (cancer cells). Changes in DNA histogram of keratin positive cells were classified into three types: Type I: the same diploid type as whole cells, Type II: the same aneuploid type as them, and Type III: the different aneuploid type as them. Among 22 cases of malignant tumor there were 3 cases of Type I, 6 cases of Type II, and 13 cases of Type III. This method could contribute greatly to the detection of cancer cells and DNA diagnosis, and thus be useful clinically.
The modulation of paraquat toxicity by tungsten was studied in vitro using cultured MDCK epithelial cells. MDCK cells were cultured in minimal essential medium with or without 1 ppm tungsten. Proliferation of cells cultured with tungsten was not inhibited after exposure to 0.25 mM or 0.5 mM paraquat. In addition, lactate dehydrogenase release into the culture medium was lower for tungsten-treated cells than for cells cultured without tungsten. Cells cultured in medium alone showed reduced viability compared with controls after exposure to 0.5 mM paraquat, but 0.25 mM paraquat did not decrease cell viability. Tungsten-treated cells showed no decrease viability in after exposure to either concentration of paraquat. Cells exposed to paraquat developed a honeycomb morphology with scanty cytoplasm and abnormal nucleoli. However, these major structural changes were not observed in cells cultured with tungsten. Our study showed that cell damage after paraquat exposure was modulated by addition of tungsten to the culture medium. It is suggested that cytosolic xanthine oxidase activity was reduced by tungsten, leading to less production of superoxide and other radicals and thus conferring resistance to paraquat toxicity.
With the recent advance in skull base surgery, approaches to the cavernous sinus and frontal skull base have been established and introduced positively. The author has applied extensive transbasal approach to lesions in the frontal skull base, and transzygomatic approach to those in the middle skull base, but since the fact that the internal carotid artery runs inside the temporal bone restricts surgical procedure for lesions extending not only to the middle skull base but to the infratemporal fossa, preservation of the internal carotid artery during surgery is demanded. Radical surgery was performed for 5 patients with malignant tumor occupying the infratemporal fossa, resulting in successful en-bloc resection of the tumor by securing a substantial operative space during surgery after early opening of the carotid canal for preservation of the internal carotid artery. In opening the carotid canal, the carotid canal triangle is first identified, then its bone is shaved and cut as far as the spine of the sphenoid, allowing a look at the inside of the tumor, or a view of the internal carotid artery, that permits en-bloc resection of the tumor. Representative cases are reported here and microanatomical information for surgical procedure in opening the carotid canal is also presented with the relevant literature.
The present report describes a case of colon cancer which presented with a rare complication of splenic abscess. A 52-year-old Japanese man with diarrhea, fever and chills was admitted to our hospital. He complained of fever, with chills at night, and abdominal pain occurring during the last month. The origin of the fever was investigated, and Escherichia coli grew from a blood culture. Multilocular splenic abscesses and wall thickening of the descending colon were revealed by CT scan, magnetic resonance imaging and ultrasound. A cancer of the descending colon was found by barium enema and colonoscopy. A curative resection was performed and the pathological report revealed the splenic abscess to have developed from a direct extension of, and perforation by, the carcinoma of the descending colon.
We investigated the relationship between scalp electroencephalogram (EEG) and nasopharyngeal EEG using Automatic EEG Analysis System in 10 patients with the age between 17 and 66 who were clinically presumed brain death because of flat line EEG and loss of auditory brainstem response (ABR). Among 10 patients, nasopharyngeal lead EEG showed low voltage and slow waves of 10-20 microV in 4 cases and flat line EEG in 6 cases. However, Automatic EEG Analysis system-assisted analysis of EEGs recorded on a magnetic tape revealed that equivalent electric potentials in the scalp EEG from these patients were smaller than those of healthy volunteers, but those in the nasopharyngeal EEG from all patients showed low voltage within the range of distribution in normal volunteers (delta:6, theta 1:6, theta 2:2, alpha 1:1, alpha 3:10, beta 1:6 in 10 cases). The present results show that in addition to the routine scalp EEG, it is clinically useful to employ simple and easy nasopharyngeal lead EEG method and to analyze data using Automatic EEG Analysis System for the judgement of brain death.