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Biomedical subjects

K Yanagihara

Publications and source records attributed to K Yanagihara.

At least 127 records · Page 7Linked to original sources
PubMed1993

Establishment and characterization of a human gastric scirrhous carcinoma cell line in serum-free chemically defined medium.

We have established a human gastric scirrhous carcinoma cell line (designated as HSC-43) in a serum-free chemically defined medium (CDM) without any polypeptide growth factor, from a primary tumor of a 56-year-old male patient. HSC-43 cells grew in vitro in adherence with a population doubling time of 55 hr, and had the cytological properties of mucinous epithelial tumor cells. Cytogenetic analysis of the cells revealed pseudotetraploidy, with structural abnormalities of deletion at chromosome Iq25 and with 3 marker chromosomes. Some cells had retained features of signet-ring cells and caused fibroblastic proliferation when transplanted into athymic nude mice. The possible involvement of transforming growth factor-alpha (TGF-alpha), and its receptor, the epidermal-growth-factor receptor (EGFR), on the growth of HSC-43 cells was studied. Synthesis and secretion of TGF-alpha by HSC-43 cells were confirmed by biological assay and enzyme-linked immunosorbent assay. Radioreceptor analysis showed the presence of receptors for EGF in HSC-43 cells. Proliferation of HSC-43 cells was inhibited by antibodies against TGF-alpha and/or the EGFR. However, neither TGF-alpha nor epidermal growth factor (EGF) was effective in stimulating the cell growth of HSC-43 cells, irrespective of the cell density when supplemented exogenously. Our data suggest that TGF-alpha and EGFR play a role in the autocrine growth of HSC-43 cells. This may be another example of growth regulation of gastric carcinoma.

Adenocarcinoma, Scirrhous↗
PubMed1993

[Local resection of pulmonary lesions by Nd-YAG laser without suturing].

Local excision of 67 pulmonary lesions in 5 patients was performed with the Nd-YAG laser. There were 2 women and 3 men with an average age of 68 years. The patients consisted of three cases of metastatic lung cancer (65 lesions), one hamartoma (One lesion) and one organized pneumonitis (one lesion). The lesion was first enucleated by a contact method using an Nd-YAG laser scalpel, then the crater of the resected lesion in the normal lung tissue was coagulated with a defocused laser beam. The craters of 52 of the 67 resected lesions which were shallow seated in the lung were left open, but the other 15 resected lesions were closed with sutures. Total blood loss was less than 100 ml. There was no bleeding or air leakage (30 cm water pressure) during the post operative days in 4 of the 5 patients, but one patients (one lesion) had to be treated by bronchoscopic endobronchial embolization against a persistent air leak. This lesion was 2.5 cm in diameter and 4 cm deep in the lung on CT. The other lesions were also measured by CT. The tumor size was 1.4 +/- 0.7 cm and the depth was 2.4 +/- 1.0 cm. The Nd-YAG laser is very suitable for local resection of peripheral lung lesions without suturing.

Aged↗
PubMed1993

Raf revertant cells resist transformation by non-nuclear oncogenes and are deficient in the induction of early response genes by TPA and serum.

A revertant cell line was generated from v-raf transformed NIH/3T3 fibroblasts. These cells, termed CHP25, express a functional v-raf oncogene. However, they are non-tumorigenic, do not form colonies in soft agar and possess a flat morphology. CHP25 cells are resistant to re-transformation by sis, ras, tyrosine kinase- as well as serine/threonine kinase-encoding oncogenes suggesting that Raf functions downstream of most membrane associated signal transducers. In contrast to v-raf transformed cells, in which the endogenous Raf-1 protein kinase is constitutively activated, v-Raf in CHP25 cells does not activate endogenous Raf-1 kinase. Since mitogen regulation of Raf-1 kinase in CHP25 cells is intact, we conclude that CHP25 cells are blocked at the level of Raf-1 substrate phosphorylation. Consistent with this interpretation CHP25 cells show specific alterations of early gene induction. The serum induction of c-fos and junD as well as the serum and TPA (12-O-tetradecanoylphorbol-13-acetate) induction of junB and egr-1 are almost completely abolished. Only v-fos can transform CHP25, whereas c-fos, v-myc, c-jun and junB are ineffective. These data suggest that the lesion responsible for the revertant phenotype of CHP25 cells is the inability to activate the AP-1 complex. We conclude that Raf-1 signaling is essential for transformation of NIH/3T3 cells by peripheral oncogenes and for regulation of a subset of early response genes by TPA and serum growth factors.

3T3 Cells↗
PubMed1992

Transforming growth factor beta 1 induces apoptotic cell death in cultured human gastric carcinoma cells.

Transforming growth factor beta 1 (TGF-beta 1) is a potent growth inhibitor for many cell types, including tumor cells. We recently have reported the establishment and characterization of two human gastric scirrhous carcinoma cell lines, HSC-39 and HSC-43. Here we examined the effect of TGF-beta 1 on the growth of these lines as compared to five other human gastric adenocarcinoma cell lines. Proliferation of HSC-39 and HSC-43 cells was strongly inhibited by TGF-beta 1, whereas the other gastric adenocarcinoma cell lines were unresponsive to TGF-beta 1. Both HSC-39 and HSC-43 cells gradually lost viability following exposure to TGF-beta 1. This response was dose dependent up to 4 ng/ml. When TGF-beta 1 was removed, the cells failed to exhibit regrowth, indicating an irreversible growth-inhibitory effect of this agent, leading to cell death. DNA fragments were observed consisting of multimers of approximately 180 base pairs 24 h after TGF-beta 1 treatment. The chromatin condensation of each cell line was confirmed by Hoechst 33258 fluorochrome staining. Ultrastructurally, condensed and fragmented nuclei were observed in TGF-beta 1-treated cells. These features are generally associated with apoptotic processes. Both cell death and DNA fragmentation were partially inhibited by cycloheximide, suggesting the requirement for new protein synthesis. Our results suggest that TGF-beta 1 induces cell death in human gastric scirrhous carcinoma cells in vitro which is mediated by activation of a signal transduction pathway for apoptosis.

Adenocarcinoma, Mucinous↗
PubMed1992

Regioselective halogenation of some 2,3-anhydro pyrimidine nucleosides with dilithium tetrahalocuprates.

The reaction of 1-(2,3-anhydro-5-O-trityl-beta-D-lyxofuranosyl)-2-O-methyluracil (1a) and its thymine analogue (1b) with dilithium tetrahalocuprate (Li2CuX4) revealed excellent to perfect regioselectivity, yielding 2,2'-anhydro-3'-halonucleosides (2a-d), while the same reactions with 2,3-anhydro uracil and thymine nucleosides (4a,b) gave arabinosyl (5a-d) and xylosyl halohydrins (6a-d) with the respective product ratio of 7:3 to 8:2. compounds 5 and 6 were isolated as the 2-O-(7) and 3- O-mesyl derivatives (8).

Bromides↗
PubMed1992

[A case report of radiation osteomyelitis 9 years after irradiation for thymoma].

Radiation osteomyelitis of the sternum is rare and usually difficult to cure. A 75-year-old man, who had undergone an exploratory sternotomy for a mediastinal tumor, not resected after all, 9 years earlier and received radiation therapy successively for the histological diagnosis of malignant thymoma, was admitted to our hospital with the chief complaint of fever and pus discharge of the anterior chest wall. He also suffered from diabetes mellitus. The skin around the fistula was dark-red and atrophic due to irradiation dermatitis and the manubrium was fissured in the midline. Open drainage and two-stage operation of direct closure was tried in vain. This case was treated successfully by resection of necrosed portion of sternum and pectoral muscle flap closure.

Aged↗
PubMed1992

[A case of lung cancer in a patient with von Recklinghausen's disease].

A 57-year-old man was admitted to our hospital for further investigation of an abnormal shadow in his chest X-ray. He had cafe-au-lait spots and multiple subcutaneous neurofibromas and was diagnosed as having von Recklinghausen's disease. Bronchofiberscopy was performed, but an adequate specimen was not obtained. Therefore, percutaneous needle biopsy was performed, and the specimen showed poorly-differentiated adenocarcinoma. The left subclavian artery was deviated on aortography, therefore, neoadjuvant chemotherapy with CDDP & VDS was performed. He subsequently underwent left upper lobectomy. Pathologically, the tumor cell showed necrosis and scarring. Including this case, there have been 11 reports of von Recklinghausen's disease associated with lung cancer in the Japanese literature. Adenocarcinoma was observed in 72.9% of cases, and poorly-differentiated tumor was observed in 7 out of 8 patients with distinct tumor differentiation.

Adenocarcinoma↗
PubMed1991

Establishment and characterization of human signet ring cell gastric carcinoma cell lines with amplification of the c-myc oncogene.

Two unique human signet ring cell gastric carcinoma cell lines (designated HSC-39 and HSC-40A) were established in vitro from the ascites of a 54-year-old male patient. Both cell lines were biologically quite similar, grew in vitro in suspension with a population doubling time of 28-30 h, and had cytological features of mucinous epithelial tumor cells. They formed colonies in soft agar, with a cloning efficiency of 0.8-1.0%. Ultrastructurally, numerous granules were observed in the cytoplasm, suggesting secretory activity. The frequent presence of desmosome and the tight junction at the cell boundary certifies the epithelial origin of the lines. Immunocytochemistry and radioimmunoassay showed production of tumor marker antigens (carcinoembryonic antigen, CA 19-9, and sialyl-Lex-i) and gastrin in both lines. These lines were transplantable in athymic BALB/c nude mice. The histopathology of each line growing in athymic BALB/c nude mice was similar to that of the original tumor. The karyotype of the cells was highly aberrant with structural and numerical changes. The presence of numerous double minute chromosomes and loss of the 13 chromosome and Y-chromosome characterize these lines. In addition, the amplified c-myc oncogene (16-32-fold) was found in both cell lines and original ascitic tumor cells. Overexpression of the c-myc mRNA was noted. These cell lines may be a useful tool, providing both in vivo and in vitro systems for further studies of the biology and therapy of human signet ring cell (or Borrmann's type IV carcinoma) gastric carcinoma.

Adenocarcinoma, Mucinous↗
PubMed1990

[Experimental studies on morphological changes of microvascular architecture following the free gingival autograft on denuded alveolar bone].

The purpose of the present studies was to examine the healing process following the free gingival autograft placed on the recipient bed either with or without periosteum in 54 adult mongrel dogs with healthy periodontium. A recipient bed was prepared on denuded alveolar bone in a definite portion of the attached gingiva of the right maxillary canine tooth, and the graft was taken from the attached gingiva of the left maxillary canine and transplanted in the recipient bed. Morphological changes were observed by means of vascular corrosion casts on the postoperatively 3rd, 5th, 14th, 21st, 28th, 42nd, 56th and 84th day. The healing process following the free gingival autograft on the denuded alveolar bone showed that this graft survived in its margin by recirculation from the cut margin of the recipient bed, and in its center the necrotic tissue changed to granulation tissue, which gradually cicatrized. This was different from the healing process following the free gingival autograft on periosteum, in which the graft was survived entirely by recirculation from the vascular plexus of the periosteum on the recipient bed. This may help to restore the function proper.

Alveolar Process↗
PubMed1990

[A case of mediastinitis caused by rupture of esophageal diverticulum].

A 42-year-old male student from abroad visited a department of internal medicine of a hospital because of fever and dyspnea. A diagnosis of purulental mediastinitis with empyema was made. After drainage of the right thoracic cavity, the patient was referred to our department. Although consecutive treatment consisting of administration of antibiotics and thoracic drainage was conducted, dyspnea became worse, and widening of the mediastinum become severer. Mediastinal drainage was done via the parasternal approach under general anesthesia. Thereafter, the patient temporarily suffered from respiratory failure, and controlled respiration by a respirator was needed. Since the amount of discharge from the mediastinal and thoracic drains gradually decreased, and clinical symptoms disappeared, the drain tubes were removed. During the clinical course, laryngoscopy, bronchoscopy, esophago-gastro-endoscopic examination and esophagography were performed. Esophagoscopy revealed a diverticulum, and esophagography showed a fistula from the esophageal diverticulum to mediastinum. From these examination results, a diagnosis of mediastinitis caused by rupture of esophageal diverticulum was established.

Adult↗
PubMed1990

[Septic pulmonary emboli caused by parenteral nutrition catheter infection].

A case of septic pulmonary emboli due to parenteral nutrition catheter infection was reported. Characteristic radiologic features were recognized. A 50-year-old man, who was receiving parenteral nutrition after total gastrectomy, consulted our department with complaints of fever and general malaise. A chest radiograph showed scattered ill-defined small peripheral nodules, which were not present before parenteral nutrition, and these nodules were quickly formed cavities + in 2nd day. He was suffering from high fever, hemo-sputum and dyspnea after removal of the parenteral nutrition catheter. Pseudomonas aeruginosa was isolated from the tip of parenteral nutrition catheter and sputum cultures. Septic pulmonary emboli were diagnosed and antibiotic therapy was performed. Bacterial endocarditis and septic thrombophlebitis were ruled out. The multiple cavity nodules extended to involve the peripheral areas of the lung and invasive shadows appeared on the chest radiograph in 8th day. Then, the invasive shadows disappeared and the walls of the cavitary lesions became thinner. After 2 months, all cavitary lesions disappeared with only linear shadows remaining.

Catheterization, Central Venous↗
PubMed1990

Isolation of a new class of 'flat' revertants from ras-transformed NIH3T3 cells using cis-4-hydroxy-L-proline.

A new class of nontransformed revertant cells has been isolated from the ras-transformed cell line DT using cis-4-hydroxy-L-proline (CHP) as a selective agent. The new revertants, CHP 9CJ and CHP CB4, each contain two copies of the v-Ki-ras gene, elevated levels of phosphorylated p21ras protein, and rescuable transforming virus, indicating that the revertant phenotype observed in these cells does not result from inactivation of v-Ki-ras or inhibition of its expression. Both CHP 9CJ and CHP CB4 revertants show a greatly reduced ability to form colonies in soft agar and to produce tumors in syngeneic mice. CHP 9CJ cells are resistant to retransformation by ras and by additional oncogenes that do not encode tyrosine kinases. A comparison of oncogene resistance patterns in these CHP-derived revertants with those from our original ouabain-derived revertants fos C-11 and F-2 indicates that oncogenes may be divided into four general groups. Oncogenes that encode proteins structurally related to p21ras comprise the first group. The second group contains only tyrosine kinase-encoding oncogenes. The third group is composed of 'nuclear', e.g. fos, and 'cytoplasmic' serine-threonine-encoding oncogenes such as mos and raf. The fourth group contains the oncogenes sis and fms.

Animals↗
PubMed1989

The relationship between connective tissue and its microvasculature in the healthy dog gingiva.

The morphological consistency of subepithelial connective tissue and its vascular architecture was studied in the healthy gingiva of adult mongrel dogs. Corrosion-cast specimens of the vasculature and specimens of well-separated connective tissue from the lamina propria were observed by SEM. Specimens injected with india ink also were examined employing light microscopy. Summarizing the results: 1) Beneath the buccal gingival epithelium, short conical papillae were seen in the subepithelial connective tissue. The capillary structure throughout this region was basically of the loop type, the overall appearance of which replicated the papillae. The papilla disappeared at the muco-gingival junction, so that after the transition to the alveolar mucosa, no papillary formation was observed. The vasculature had transformed from loop to network structure. 2) Beneath the sulcular gingival epithelium, the subepithelial connective tissue displayed a smooth concavity with no papillae. In this region, the capillaries formed a flat, dense network in accordance with the morphology of the connective tissue. The capillary network suddenly disappeared at the terminal edge of the inner epithelium. The vessels of the lamina propria located immediately beneath the network reduced their lumina at the margin and formed the plexus of the periodontium, displaying a rather open network.

Acrylic Resins↗
PubMed1989

Periosteal microvasculature in the dog alveolar process.

Most periodontal surgery has been performed on the basis of the regenerating capacity of the periosteum. Recently it has been pointed out that the blood supply is important to the osteogenic and fibrogenic activity of the periosteum. The purpose of the present study was three-dimensional observation of blood vessels distributed within the gingival periosteum and the periosteum beneath the alveolar mucosa of adult mongrel dogs. Corrosion cast specimens of vessels were observed from three directions: mucosal side, bone side, and horizontal cut surface. It was revealed that the plexus distributed in the periosteum of the gingiva formed a coarse network structure, which consisted mainly of arterioles and venules. In contrast, in the periosteum of the alveolar mucosa, a dense network structure consisting of arterioles, capillaries, and venules formed a vascular bed. Thus, the mucogingival junction was easily located even in the plexus of periosteum which was distributed beneath the gingiva and the alveolar mucosa, suggesting that the difference in tissue specificity was reflected in the plexus of the periosteum.

Alveolar Process↗
PubMed1988

Differential growth sensitivity to 4-cis-hydroxy-L-proline of transformed rodent cell lines.

The effect of 4-cis-hydroxy-L-proline (CHP), a proline analogue, on the anchorage-dependent and -independent growth of several transformed rodent cell lines was studied. Mouse NIH-3T3 fibroblasts transformed by a variety of different oncogenes (Ki-ras, mos, src, fms, fes, met, and trk) by a DNA tumor virus (SV40) or by a chemical carcinogen (N-methylnitrosourea) were all found to be more sensitive (50% inhibitory dose, 20 to 55 micrograms/ml) to the dose-dependent inhibitory effects of CHP on growth in monolayer culture than were NIH-3T3 cells (50% inhibitory dose, 120 micrograms/ml). CHP was generally found to be even more effective in inhibiting the growth of these transformed cells as colonies in soft agar than in monolayer cultures. In addition, rat embryo fibroblasts (CREF) and normal rat kidney fibroblasts (NRK) after transformation with a Ki-ras oncogene exhibit a similar increase in their sensitivity to CHP-induced growth inhibition. Treatment of NRK cells with transforming growth factor alpha (TGF-alpha) and beta (TGF-beta), which reversibly induces phenotypic transformation of these cells, increases their sensitivity to CHP to a level comparable with that observed in Ki-ras-transformed NRK cells (K-NRK). The growth inhibitory effects of CHP are reversible, since removal of CHP results in a normal resumption of cell growth. CHP uptake occurs primarily through the Na+- and energy-dependent neutral amino acid transport A system, which is 6- to 7-fold more elevated in K-NRK cells compared with NRK cells. Treatment of NRK cells with TGF-alpha and/or -beta increases the uptake of [3H]methylaminoisobutyric acid on the A system to a level that is similar to that found in K-NRK cells. The functions of the Na+/K+ and Na+/H+ exchange systems are apparently necessary for the enhanced A system activity, since ouabain and amiloride can inhibit the uptake of [3H]methylaminoisobutyric acid in K-NRK cells and in NRK cells treated with TGF-alpha and/or -beta. The activity of the A system is specifically increased in K-NRK and in TGF-alpha- and/or -beta-treated NRK cells, since the other two major neutral amino acid uptake systems, the ASC and the L systems, and the Ly+ system for basic amino acid uptake show no apparent changes in their activity in NRK cells after treatment with TGF-alpha and/or -beta or in these cells after transformation with the Ki-ras oncogene. These results suggest that the differential growth sensitivity to CHP of transformed rodent cells and of normal fibroblasts treated with TGF-alpha and/or -beta is due in part to an elevated uptake of this amino acid analogue on the neutral amino acid transport A system.

Amiloride↗
PubMed1988

A system for deriving revertants of oncogene-transformed human cells.

We have previously shown that some transformed derivatives of the human osteosarcoma-derived cell line HOS are killed by treatment with 1 microM ouabain at pH 8.2, whereas their nontransformed counterparts are relatively unharmed by the same conditions. HOS cells transformed by v-Ki-ras and RAS, v-fms, or MET are susceptible to 1 microM ouabain while those transformed by v-fes are not. Here we describe the adaptation of this differentially cytotoxic effect as a method to enrich for cells which revert to a nontransformed phenotype. We have optimized parameters which increase the differential cytotoxicity, including pH and potassium concentration during and subsequent to ouabain treatment. The efficiency of this procedure was tested in mixed cell experiments where model populations were constructed consisting of HOS cells mixed with an excess of v-Ki-ras-transformed HOS cells. Two successive OAK treatments (ouabain/alkaline/K+-free) were sufficient to recover nontransformed cells free of ras-transformants as indicated by genetic markers and morphology. This HOS/ouabain system is currently being used to derive revertants of ras-transformed human cells and could facilitate the isolation of genes interacting in the pathways by which these cells are transformed.

Cell Line, Transformed↗