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Y Sasaguri

Publications and source records attributed to Y Sasaguri.

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

Immunopathology and cytokine detection in the skin lesions of patients with Kawasaki disease.

To investigate the pathogenesis of Kawasaki disease, we examined biopsy specimens from the skin lesions of 10 Japanese patients at the site of the polymorphous exanthem on the hip and at the site of an inoculation with BCG (bacillus Calmette-Gruérin) vaccine of two of them. Ten normal patients were also examined for control purposes. The histopathologic features of the skin lesions were characterized by extensive edema with dilation of the small blood vessels in the papillary dermis; the site of BCG vaccine inoculation showed much greater inflammatory change; neutrophil emigration was weak and most of the infiltrates were CD4+ T lymphocytes and CD13+ macrophages. The expression of the DR locus of human leukocyte antigen was detected not only on the epidermal keratinocyte surface but also on the walls of the small blood vessels and the infiltrating cells around these blood vessels. We also detected cytokines in the skin lesions: (1) interleukin-1 alpha and tumor necrosis factor alpha were strongly positive in all patients with acute Kawasaki disease, (2) interleukin-2 and interferon gamma were weakly or partially positive, (3) no cytokines were detected in the convalescent phase, and (4) the amounts of cytokines at the site of BCG vaccine inoculations were larger than those at the site of the polymorphous exanthem. These findings suggest that CD4+ T lymphocytes and CD13+ macrophages are activated and interleukin-1 alpha and tumor necrosis factor alpha may be involved in the pathogenesis of the inflammation of acute Kawasaki disease.

BCG Vaccine↗
PubMed1993

Production of matrix metalloproteinase 9 (92-kDa gelatinase) by human oesophageal squamous cell carcinoma in response to epidermal growth factor.

We demonstrated that four human oesophageal squamous cell carcinoma cell lines (TE8, TE9, TE10 and TE11) produced matrix metalloproteinase-1 (proMMP-1/tissue collagenase), 2 (ProMMP-2/'type IV collagenase'), 3 (proMMP-3/stromelysin), and 9 (proMMP-9/92-kDa gelatinase) as members of a matrix metalloproteinase (MMP) family, which degrades extracellular matrix macromolecules. Under normal culture conditions, in immunoblot analysis, proMMP-1 of M(r) = 53,00 was detected in one cell line (TE8), proMMP-2 of M(r) = 72,000 in three cell lines (TE9, TE10, and TE11), and proMMP-3 of M(r) = 57,000 in all four cell lines. In addition to these enzymes, in enzymography, a gelatinolytic activity around M(r) = 92-kDa, likely to be proMMP-9, was detected in only one cell line (TE10) under normal culture conditions. When these cell lines were treated with epidermal growth factor (EGF), however, the agent stimulated three cell lines (TE8, TE10 and TE11) to produce proMMP-9 in a dose-dose dependent manner. Oesophageal carcinoma-conditioned medium stimulated oesophageal fibroblasts to produce proMMP-1, -2, and -3, suggesting that the interaction between oesophageal carcinoma and stromal fibroblasts also plays a role in the production of MMPs by the latter. Our present study illustrates that oesophageal squamous cell carcinoma produces a variety of MMPs including proMMP-1, -2, -3, and -9 in vitro, suggesting that the ability of MMP production of the tumour may play an important role in its malignant behaviour and that the production of proMMP-9 may be regulated by EGF via overexpression of EGF receptors.

Carcinoma, Squamous Cell↗
PubMed1993

Expression of matrix metalloproteinase-2 related to lymph node metastasis of oral squamous cell carcinoma. A clinicopathologic study.

To investigate the effect of matrix-degrading enzymes on the malignant potential of oral squamous cell carcinoma, the expression of matrix metalloproteinase-2 (MMP-2/72-kD gelatinase/type IV collagenase) in 46 patients who had neck surgery for oral cancer was studied immunohistochemically. In 20 of 26 patients (76.9%) with lymph node metastases, proMMP-2 was strongly expressed, whereas the production of proMMP-2 in tissue was detected only in 5 of 20 patients (25%) who had no lymph node metastases. In tissue specimens, proMMP-2 was expressed in a diffuse invasive mode and in the advancing front of cancer. Because MMP-2 can degrade type IV collagen composed of basement membrane, these results suggest that the in vivo production of the enzyme by cancer is an indicator of the degree of malignancy, and that the analysis of proMMP-2 expression is useful to evaluate the malignant potential in individual oral squamous cell carcinoma.

Adult↗
PubMed1993

Porcine vascular smooth muscle cells immortalized with SV40 ori-defective DNA: characteristics of cell growth and collagen synthesis.

A cell line derived from medial smooth muscle cells (SMC) was established from the porcine coronary artery by transfection with ori-defective simian virus 40 plasmid DNA (SV40 DNA). The characteristics of transfected cells (SV40-SMC) such as cell growth, collagen and non-collagen syntheses were investigated. SV40-SMC expressed SV40 large T antigen, c-myc and c-myb encoded proteins in the nuclei. SV40-SMC demonstrated a 'hills and valleys'-like arrangement in overconfluence and actin filaments upon immunofluorescent staining. Under electron microscopic observation, SV40-SMC had larger amounts of synthetic organelles and smaller amounts of filament bundles than those of SMC. SV40-SMC demonstrated three times higher growth activity and 4.4 times greater cellular density than SMC. Smooth muscle cells did not grow in media containing 5% plasma derived serum (PDS) instead of normal serum, whereas SV40-SMC proliferated in this medium. SV40-SMC did not grow in soft agar gel, while HeLa S3 cells, a cell line of human cervical carcinoma, formed colonies in this gel. By immunofluorescent (IF) staining, collagen phenotypes I, III, IV and V were detected in both SV40-SMC and SMC. However, protein synthesis including collagen and non-collagen was higher in SV40-SMC than in the control sample. It was concluded that SV40-SMC were a continuous cell line for vascular SMC regarding morphological characteristics, and demonstrated a higher growth activity, with increased collagen and non-collagen syntheses. This cell line is useful for the investigation of atherogenesis in relation to a proliferation of SMC and an accumulation of extracellular matrices in vascular intima.

Agar↗
PubMed1993

Induction of gelatinolytic neutral proteinase secretion by lipid peroxide in cultured mesangial cells.

To investigate whether lipid peroxides in tissue and/or blood play an important role in the injury to mesangial cells and glomerular structural proteins, we studied the effect of linoleic acid hydroperoxide (LHO) on mesangial cells cultured from rat kidneys. When 1.0-3.0 nmol/ml of LHO were added to the culture, the production of 70-kDa gelatinolytic neutral proteinase by mesangial cells was enhanced without morphological damage. The production of the enzyme, however, was inhibited by an addition of 4.0 or 5.0 nmol/ml of LHO and a microscopic examination also demonstrated marked injury to mesangial cells in the presence of these concentrations of LHO. These data suggest that there is a possibility that increased lipid peroxides in tissue and/or blood play a role in the injury to mesangial cells and glomerular structural proteins leading to mesangiopathy.

Animals↗
PubMed1993

Down-regulation in the production of matrix metalloproteinase 1 by human aortic intimal smooth muscle cells.

Effects of dexamethasone, retinoic acid, prostaglandin E2 (PGE2), and Iloprost as a agonist of prostacyclin (A-PGI2) on DNA synthesis and production of a precursor of matrix metalloproteinase 1 (tissue procollagenase/proMMP-1) by human aortic smooth muscle cells were investigated. When after treatment with platelet-derived growth factor (PDGF), these agents were added to the cultures, DNA synthesis and production of proMMP-1 were inhibited in a dose-dependent manner. These results suggest that these agents are negative regulators of PDGF. Since these agents are present in the blood or produced in the blood wall, in addition, since PDGF plays the most important role in the process of atherosclerosis, we propose that these agents function in vivo as a systems of protection against atherosclerosis.

Aorta↗
PubMed1993

Simple bone cyst. Treatment by trepanation and studies on bone resorptive factors in cyst fluid with a theory of its pathogenesis.

Simple bone cysts were treated by trepanation. The technique consists of drainage of cyst fluid, lavage of the cystic cavity with saline, and the making of multiple drilling holes through the cortical and the medullary bone of the cyst wall. Injection of corticosteroid was omitted. In 11 cases treated by this method, the clinical outcome was good. Biochemical analyses of the cyst fluid showed bone-resorptive factors, i.e., prostaglandins, interleukin 1, proteolytic enzymes. Electrophoretic analysis of proteolytic enzymes in polyacrylamide gel containing sodium dodecyl sulfate and polymerized gelatin showed proteins with molecular weights of about 130,000, 92,000, 72,000, and lower than 50,000. Increase in such bone-resorbing activities seems to be one of the causative factors in simple bone cysts. The technique was effective in decompressing the internal pressure of the cysts, improving the blood flow through the medullary bone of the cyst wall, stimulating the periosteum to induce bone formation, and eliminating bone destruction.

Adolescent↗
PubMed1992

Production of matrix metalloproteinase-2 and metalloproteinase-3 related to malignant behavior of esophageal carcinoma. A clinicopathologic study.

BACKGROUND: Matrix metalloproteinases (MMP) are a gene family of zinc enzymes capable of degrading almost all of the extracellular matrix macromolecules in vivo. Their enzymic activities are believed to be responsible for tumor invasion and metastasis. METHODS: In this study, using peroxidase-antiperoxidase method, monospecific antisera against MMP-1 (tissue collagenase), MMP-2 (type IV collagenase/72-kilodalton [KD] gelatinase), and MMP-3 (stromelysin) were applied to 29 squamous cell carcinomas and normal epithelium of the esophagus to identify cells synthesizing and secreting these enzymes. RESULTS: Immunoreactivity of MMP-1, -2, and -3 was observed in small cancer nests of the deeply invasive or marginal portion of the tumor. Among the 29 patients studied, the presence of at least one MMP was observed in 17 (58.6%). All three enzymes were observed in six (20.6%) patients, MMP-2 and -3 in five (17.2%) patients, only MMP-2 in three (10.3%) patients, and MMP-3 alone in three (10.3%) patients. There was a good correlation among histologic stage and tumor invasion, lymph node metastasis, and MMP expression. In particular, expression of MMP-2 and -3 was closely related to lymph node metastasis and vascular invasion. CONCLUSIONS: These results suggest that MMP, especially MMP-2 and -3, play an important role in tumor invasion and metastasis and that analysis of MMP-2 and -3 production is useful for evaluation of malignant potential in esophageal carcinoma.

Aged↗
PubMed1992

Purification and characterization of matrix metalloproteinase 9 from U937 monocytic leukaemia and HT1080 fibrosarcoma cells.

The precursor of matrix metalloproteinase 9 (proMMP-9), also known as '92 kDa progelatinase/type IV procollagenase', was purified from the conditioned medium of U937 monocytic leukaemia and HT1080 fibrosarcoma cell lines stimulated with phorbol 12-myristate 13-acetate. ProMMP-9 in these culture media is non-covalently complexed with the 29 kDa tissue inhibitor of metalloproteinases (TIMP), but free proMMP-9 was separated from the TIMP-proMMP-9 complex by chromatography on Green A Dyematrex gel. The final product was homogeneous on SDS/PAGE, with a molecular mass of 88 kDa without reduction and 92 kDa with reduction. Treatment of proMMP-9 with 4-aminophenylmercuric acetate converted the 88 kDa precursor into 80 kDa and 68 kDa forms. Gelatin-containing zymographic analysis showed zones of lysis associated with all three species. However, only the 68 kDa species was shown to be catalytically active by its ability to bind to alpha 2-macroglobulin. In the presence of an equimolar amount of TIMP, only the 80 kDa species was generated by treatment with 4-aminophenylmercuric acetate, but no enzyme activity was detected. This indicates that TIMP binds to the 80 kDa intermediate and inhibits the generation of the active 68 kDa species. Eight endopeptidases (trypsin, chymotrypsin, plasmin, plasma kallikrein, thrombin, cathepsin G, neutrophil elastase and thermolysin) were tested for their ability to activate proMMP-9. Of them, trypsin was the most effective activator of proMMP-9. Only partial activation (10-30%) was observed with plasmin, cathepsin G and chymotrypsin. The active forms generated by trypsin were identified as 80 kDa, 74 kDa and 66 kDa by their abilities to bind to alpha 2-macroglobulin. In the presence of an equimolar amount of TIMP, proMMP-9 was also converted into the same molecular-mass species by trypsin, but they were not proteolytically active. This suggests activated MMP-9 is inhibited by TIMP. Activated MMP-9 digested gelatin, type-V collagen, reduced carboxymethylated transferrin and, to a lesser extent, type-IV collagen and laminin A chain. The specific activity against gelatin was estimated to be 15,000 units/mg (1 unit = 1 microgram of gelatin degraded/min at 37 degrees C) by titration with alpha 2-macroglobulin. Comparative studies on digestion of gelatin and collagen types IV and V by MMP-9 and MMP-2 indicated that both enzymes degrade these substrates into similar fragments. However, the susceptibilities of laminin, fibronectin and reduced carboxymethylated transferrin to these two MMPs were sufficiently different to indicate differences in substrate specificities between these two closely related proteinases.

Electrophoresis, Polyacrylamide Gel↗
PubMed1992

Effects of interleukin-6 on the metabolism of connective tissue components in rheumatoid synovial fibroblasts.

OBJECTIVE: High levels of interleukin-6 (IL-6) have been found in the synovial fluid of patients with rheumatoid arthritis (RA). We undertook the present study to investigate the role of IL-6 in this disease. METHODS: We examined the effects of IL-6, in comparison with IL-1, on the biosynthesis of extracellular matrix macromolecules and of matrix-degrading proteinases in rheumatoid synovial fibroblasts. RESULTS: In rheumatoid synovial fibroblasts, IL-6 by itself enhanced the production of plasminogen activator, its inhibitor, and tissue inhibitor of metalloproteinases, whereas it did not modulate the biosynthesis of precursor of matrix metalloproteinase 1 (proMMP-1) (tissue collagenase), proMMP-3 (stromelysin), or connective tissue components. However, IL-1-induced production of proMMP-1 and proMMP-3 was preferentially augmented by IL-6. CONCLUSION: These results suggest that in RA, IL-6 may participate along with IL-1 in fine tuning of the catabolism of connective tissue components, by modulating the balance between connective tissue-degrading enzymes and their inhibitors.

Arthritis, Rheumatoid↗
PubMed1992

Production of matrix metalloproteinase 2 (gelatinase/type IV collagenase) and 3 (stromelysin) by cultured oral squamous cell carcinoma.

Matrix metalloproteinases (MMPs) are believed to play an important role in tumor invasion and metastasis. MMPs have been identified as proforms of malignant tumor-associated enzymes, such as procollagenase (proMMP-1) of M(r) = 53,000, progelatinase (proMMP-2) of M(r) = 72,000, proMMP-9 of M(r) = 92,000, and prostromelysin (proMMP-3) of M(r) = 59,000. Here we report that two cell lines of squamous cell carcinoma (SCC9 and SCC25) produce at least two matrix metalloproteinases (MMPs) in zymogen form, which have been identified as proMMP-2 and 3 by indirect immunofluorescence technique, immunoblot analysis, and gelatin-substrate gel enzymography. Additionally, a 92-kDa gelatinolytic metalloproteinase (proMMP-9) was detected by gelatin-substrate gel enzymography. We propose that the ability of these tumor cells to secrete MMPs plays an important role in the malignant behavior of oral squamous cell carcinomas.

Adult↗
PubMed1992

Immortalization of human aortic smooth muscle cells with origin-minus simian virus 40 DNA.

We established two cell lines of human smooth muscle cells (SMC) by transfection of cells from the aortic intima and aortic media with origin-minus simian virus 40 (ori-minus SV40) DNA. Ori-minus SV40 DNA very efficiently immortalized human smooth muscle cells in culture. Proteins that these cell lines produced included type I, III, IV, and V collagens, fibronectin, and human matrix metalloproteinases (MMP)-1 (tissue collagenase), -2 ("type IV collagenase"), and -3 (stromelysin). The protein production in these cell lines generally mimicked that of normal SMC, but the immortalization stimulated the cell line of medial SMC to produce excessive MMP-2 and to secrete MMP-9 (92-kDa gelatinase). However, since these cell lines did not show a fully malignant phenotype, we concluded that, in addition to the degradation of extracellular matrix macromolecules, including basement membrane components by MMP-2, -3, and/or -9, some additional factors must be involved for the malignancy of fully transformed cells and that these immortalized human aortic SMC, which share many characteristics with normal SMC, will prove useful to study the role(s) of metalloproteinases in atherosclerosis.

Animals↗
PubMed1992

Rapidly destructive arthropathy of the hip. Studies on bone resorptive factors in joint fluid with a theory of pathogenesis.

The mechanism of joint destruction in rapidly destructive coxopathy was studied by analyzing bone resorptive factors in the joint fluid. Prostaglandins were found to play a partial role in joint destruction. Some cases of rapidly destructive coxopathy revealed elevated levels of interleukin-1 beta (IL-1 beta) in the joint fluid. Electrophoretic analysis of proteolytic enzymes in polyacrylamide gel containing sodium dodecyl sulfate and copolymerized gelatin demonstrated that the resorptively active peptides have relative molecular weights (M(r)) of approximately 92,000, 72,000, and lower than 60,000. Cultured cells from synovia obtained perioperatively secreted matrix metalloproteinase 2 (MMP-2) with an M(r) of 72,000 and matrix metalloproteinase 3(MMP-3) with an M(r) of 57,000. Synovial cells from the patients with coxarthrosis secreted fewer proteolytic enzymes. Prostaglandins, IL-1 beta, MMP-2, and MMP-3 could act synergetically as promotors in the rapid destruction of the hip joint.

Bone Matrix↗
PubMed1992

Cell proliferation-related production of matrix metalloproteinases 1 (tissue collagenase) and 3 (stromelysin) by cultured human rheumatoid synovial fibroblasts.

We investigated the effects of platelet-derived growth factor (PDGF), epidermal growth factor (EGF), and insulin on the cell proliferation of and the production of matrix metalloproteinases (MMPs) by rheumatoid synovial fibroblasts in order to determine the role of these agents in rheumatoid arthritis. PDGF stimulated rheumatoid synovial fibroblasts to increase DNA synthesis and the production of precursor forms of MMP-1 of M(r) = 53,000 and -3 of M(r) = 57,000. EGF and insulin also increased DNA synthesis and the production of these enzymes, but the amount of DNA or MMPs was smaller than that induced by PDGF. Since the production of matrix macromolecules and their degradation is essential for the remodelling of synovial tissue in rheumatoid arthritis, these data suggest that the production of MMP-1 and-3 by rheumatoid synovial fibroblasts in relation to cell proliferation plays an important role in the pathological process of rheumatoid arthritis.

Arthritis, Rheumatoid↗
PubMed1992

Production of matrix metalloproteinases 2 and 3 (stromelysin) by stromal cells of giant cell tumor of bone.

Matrix metalloproteinases play a central role in the catabolism of extracellular matrix macromolecules. Here the authors report that giant cell tumor of bone (GCT) produces two matrix metalloproteinases (MMPs) in zymogen form, which have been identified as proMMP-2 (also known as "72-kDa-progelatinase/type IV procollagenase") and proMMP-3 (prostromelysin). Giant cell tumor is known to consist of two major cell populations, multinucleated giant cells and stromal cells. On several passages of the tumor cells in culture, only stromal cells proliferated. These stromal cells produced proMMP-2 but not proMMP-3. Addition of the conditioned medium of primary GCT culture or human macrophage-conditioned medium to the passaged stromal cells induced the production of proMMP-3. The production of proMMP-3 was also induced by interleukin 1 (IL-1), but not by tumor necrosis factor alpha (TNF alpha). ProMMP-1 (tissue procollagenase) was not detected even after treatment with these stimuli. Immunohistochemical studies have demonstrated that multinucleated giant cells in GCT both produce IL-1 and TNF alpha, suggesting that IL-1 secreted by multinucleated giant cells may be responsible for in vivo production of proMMP-3 by the stromal cells. The authors propose that GCT has a self-stimulatory system for the production of matrix-degrading proteinases and that the ability of the passaged stromal cells to synthesize and secrete proMMP-3 with appropriate stimuli may contribute the malignant behavior of GCT.

Adult↗
PubMed1991

Collagenase production by immortalized human aortic endothelial cells infected with simian virus 40.

Human aortic endothelial cells, isolated at autopsy from a 52-year-old male dying from lung cancer, were treated with simian virus 40 (SV40). One colony was isolated from the infected endothelial cell culture 4 weeks after infection. The cells expressed SV40 large T antigen and p53 protein (p53) in their nuclei but lacted the characteristics of a transformed phenotype. The cells grew well in a monolayer over the 97th passage and exhibited Factor VIII-related antigen, Ulex europaeus 1 agglutinin (UEA-1) as endothelial cell markers, and a well-developed fibronectin network. The amount of prostacyclin synthesized by the cells was less than the amount synthesized by normal aortic or umbilical cord vein endothelial cells. The cells produced relatively large amounts of procollagenase, and 12-o-tetradecanoyl-phorbol-13-acetate (TPA) augmented the ability of the cells to produce this enzyme. These immortalized human aortic endothelial cells, which have some characteristics of normal endothelial cells and, like capillary endothelial cells, have the ability to produce collagenase, will probably prove useful for studies of atherosclerosis and angiogenesis.

Animals↗