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S Inoue

Publications and source records attributed to S Inoue.

At least 721 records · Page 40Linked to original sources
PubMed1995

Diabetes impairs DRG neuronal attachment to extracellular matrix proteins in vitro.

Attachments of dorsal root ganglion (DRG) neurons from streptozotocin (STZ)-induced diabetic and normal C57BL mice to the following substrates were evaluated in vitro: a) poly L-lysine (PL), b) PL + type I collagen (CL-I), c) PL + type IV collagen (CL-IV), d) PL + laminin (LM) and e) PL + fibronectin (FN). After 6 h in culture, there was no significant difference in the average ratio of cells adhesive to PL between the diabetic (74.9%) and normal group of mice (75.6%). In the normal group, the addition of extracellular matrix (ECM) proteins such as CL-I, CL-IV, LM and FN to PL increased the ratios of cell attachment from 75.6% to nearly 90%. In the diabetic group, however, none of these proteins improved the attachment (the ratio changed from 74.9% to nearly 70%). Survival and neurite extension of attached cells after 48 h in culture were not different between the two groups. These results suggest that the cell-surface receptors, which enable DRG neurons to bind to the extracellular matrix proteins, are impaired by diabetes, resulting in being one of the causes of diabetic neuropathy.

Animals↗
PubMed1995

Clinical significance of portal invasion by pancreatic head carcinoma.

BACKGROUND: The purpose of the present study was to clarify the indication of aggressive surgery for pancreatic head carcinoma. METHODS: Laparatomy was performed in 153 patients with carcinoma, 101 of whom underwent resection of the carcinoma. With histologic examination the degree of carcinoma invasion into the portal vein was classified into grades 0, I, or II according to the depth of invasion by the carcinoma. Macroscopic carcinoma invasion into portal vein was classified into types A, B, C, or D according to preoperative findings on the portal phase of superior mesenteric angiography or intraoperative portography. RESULTS: Macroscopic findings correlated with the histologic invasion grades. The 1-year survival rate was 39.6% in grade 0, 11.3% in grade I, and 5.5% in grade II cases. The survival rates of patients with type A (p < 0.01), B (p < 0.05), and C invasion (p < 0.01) were higher than those of patients who did not undergo resection; however, no significant difference in the survival rates between patients who did not undergo resection and patients with type D invasion was observed. CONCLUSIONS: For locally advanced carcinoma of the pancreatic head or entire pancreas, patients with type D invasion have no indication of aggressive surgery.

Adult↗
PubMed1995

Role of intravascular ultrasonography in detecting intravascular tumor thrombi: a preliminary report.

BACKGROUND: We evaluated the role of intravascular ultrasonography in the diagnosis of intravascular tumor thrombi. METHODS: During the past 2 years intracaval endovascular ultrasonography was performed in 26 selected patients to diagnose inferior vena cava invasion. Results of positive intracaval endovascular ultrasonogram were correlated with the pathologic findings of resected specimens and autopsy and with other imaging technologies such as computed tomography and angiography. RESULTS: Six patients had positive studies of intracaval tumor thrombus. In all cases detailed horizontal images perpendicular to the inferior vena cava axis were studied. Five of the six patients underwent resection. Intravascular ultrasonography correctly predicted the extent of the tumor thrombus, the degree of tumor adherence to the vessel wall, and the intraluminal movement of the tumor thrombus. Floating thrombi were visualized as an intraluminal to-and-fro movement. Thrombus adhesion to the vessel wall appeared as an absence of space between the tumor and the wall, with no respiratory movement of the thrombus. CONCLUSIONS: Intravascular ultrasonography was useful for the accurate diagnosis of intravascular tumor thrombi and aided in formulating the operative strategy.

Adult↗
PubMed1995

Metal-mediated oxidative damage to cellular and isolated DNA by certain tryptophan metabolites.

The tryptophan metabolites 3-hydroxyanthranilic acid (3-HAA) and 3-hydroxykynurenine (3-HKyn) are carcinogens. DNA damage by 3-HAA and 3-HKyn in the presence of metal ions was investigated as a potential mechanism of their carcinogenicity. Pulsed field gel electrophoresis showed that in the presence of Mn(II), 3-HAA and 3-HKyn induced DNA double-strand breaks in cultured human cells. DNA single-strand breaks were observed with alkali treatment. The enhancing effect of catalase inhibitor and the inhibitory effect of o-phenanthroline on the strand breakage indicated the involvement of H2O2 and endogenous transition metal ion. Damage to DNA fragments obtained from c-Ha-rds-1 protooncogene was investigated by a DNA sequencing technique. 3-HAA and 3-HKyn induced piperidine-labile sites frequently at thymine and guanine residues in the presence of Cu(II). The inhibitory effects of bathocuproine and catalase on Cu(II)-mediated DNA damage suggest that Cu(I) and H2O2 have important roles in the production of active species causing DNA damage. The Cu(II)-mediated DNA damage was enhanced by preincubation of 3-HAA with Mn(II). UV-visible spectroscopy showed that Mn(II) and Cu(II) enhanced the rate of autoxidation of 3-HAA in different ways. These results suggest that in the presence of Mn(II) or Cu(II), these tryptophan metabolites produce H2O2, which is activated by transition metal ion to cause damage to DNA both in the case of isolated DNA and cultured cells.

3-Hydroxyanthranilic Acid↗
PubMed1995

Expression of new KDN-gangliosides in rainbow trout testis during spermatogenesis and their structural identification.

The developmental expression of 2-keto-3-deoxy-D-glycero-D-galacto-nononic acid-containing glycosphingolipids (KDN-gangliosides) in rainbow trout testis during spermatogenesis was studied using a monoclonal antibody, mAb.kdn3G, which recognizes the KDN alpha 2-->3Gal beta 1-->epitope. A major KDN-ganglioside found in mature sperm, (KDN)GM3, KDN alpha 2-->3Gal beta 1-->4Glc beta 1-->Cer (where Cer is ceramide), was expressed in testis throughout all stages of its maturation. On the contrary, four new KDN-gangliosides which were reactive with mAb.kdn3G were not detected in mature sperm, although they were identified in immature testis and expressed during spermatogenesis. The structures of these KDN-gangliosides were established by chemical, enzymatic and immunochemical methods as: (i) (KDN)GD1a, KDN alpha 2-->3Gal beta 1-->3GalNAc beta 1-->4(KDN alpha 2-->3)Gal beta 1-->4Glc beta 1-->Cer; (ii) (KDN, Neu5Ac)GD1a, KDN alpha 2-->3Gal beta 1-->3GalNAc beta 1-->4(Neu5Ac alpha 2-->3)Gal beta 1-->4Glc beta 1-->Cer and Neu5Ac alpha 2-->3Gal beta 1-->3GalNAc beta 1-->4(KDN alpha 2-->3)Gal beta 1-->4Glc beta 1-->Cer; (iii) (KDN) GD1 alpha, KDN alpha 2-->3Gal beta 1-->3(KDN alpha 2-->6)GalNAc beta 1-->4Gal beta 1-->4Glc beta 1-->Cer; and (iv) (KDN,Neu5Ac)GD1 alpha, KDN alpha 2-->3Gal beta 1-->3(Neu5Ac alpha 2-->6)GalNAc beta 1-->4Gal beta 1-->4Glc beta 1-->Cer. (KDN)GD1a and (KDN,Neu5Ac)GD1a first appeared at an early stage of spermatogenesis, but (KDN)GD1 alpha and (KDN,Neu5Ac)GD1 alpha were not expressed until 2 months prior to spermiation. While (KDN)GM3 was previously shown to contain only 4-sphingenine (d18:1) acylated with a C16:0 fatty acid, the new KDN-gangliosides discovered in this study were composed of 4-hydroxysphinganine (t18:0) or 4-sphingenine (d18:1), and were acylated with a C24:1 or C16:0 fatty acid. A possible function of these KDN-gangliosides is suggested.

Acylation↗
PubMed1995

A precise structural analysis of a fertilization-associated carbohydrate-rich glycopeptide isolated from the fertilized eggs of euryhaline killi fish (Fundulus heteroclitus). Novel penta-antennary N-glycan chains with a bisecting N-acetylglucosaminyl residue.

A novel carbohydrate-rich sialoglycopeptide of apparent molecular mass approximately 6 kDa was isolated from the fertilized eggs of Fundulus heteroclitus (euryhaline killi fish). This glycopeptide is a member of the L-hyosophorin family, characterized by its high content of carbohydrate (80-90% by weight) and formed by depolymerization of the precursor glycopoly-protein (H-hyosophorin) upon fertilization. The structures of the N-glycan chains were unambiguously established by a combination of compositional analysis, methylation analysis, selective chemical degradation (periodate oxidation-Smith degradation and hydrazinolysis-nitrous acid deamination), enzymatic (peptide:N-glycosidase F, several beta-galactosidases, beta-hexosaminidase and alpha-galactosidase) digestions and instrumental analyses (1H-NMR and fast atom bombardment mass spectrometry) to have the novel and unique carbohydrate sequences, Gal alpha 1-->3(Gal beta 1-->4)Gal beta 1-->4GlcNAc beta 1--> and Gal alpha 1-->3(+/- GalNAc beta 1-->4GlcNAc beta 1-->3Gal beta 1-->4)Gal beta 1-->4GlcNAc beta 1-->. This study represents the first detailed investigation of the nature of bulky complex asparagine-linked penta-antennary glycans with a bisecting GlcNAc residue in glycoproteins. Expression of such bulky multiantennary glycan units on proteins may be essential during early embryogenesis.

Acetylglucosamine↗
PubMed1995

Binding mode of phospholipase A2 with a new type of phospholipid analog having an oxazolidinone ring.

Inhibition of phospholipases A2 (PLA2s) by a new type of monodispersed phospholipid analog, 3-dodecanoyl-4-phosphatidylcholinohydroxymethyl-2-oxazolidinone (oxazolidinone-PC), was investigated by the pH stat assay method using monodispersed 1,2-dihexanoyl-sn-glycero-3-phosphorylcholine (diC6PC) as the substrate. The PLA2s used were those from bovine pancreas and cobra (Naja naja atra) venom (Group I) and from Japanese mamushi (Agkistrodon halys blomhoffii) venom (Group II). This new-type substrate analog was shown to inhibit competitively both types of venom and bovine pancreatic enzymes by binding to the active site in a similar manner to the carboxamide-type analog 2-dodecanoyl-amino-1-hexanol-phosphocholine (amide-PC). The binding of a stereoisomer, (R)-amide-PC, to N. naja atra (Group I) and A. halys blomhoffii (Group II) PLA2s was facilitated by the binding of Ca2+ to the enzymes. On the other hand, the binding of (R)-oxazolidinone-PC to the N. naja atra (Group I) enzyme was found to be independent of Ca2+ binding, while its binding to the A. halys blomhoffii (Group II) enzyme was markedly facilitated by the binding of Ca2+ to the enzyme. The binding of (R)-amide-PC to N. naja atra PLA2 (Group I) was markedly influenced by the ionization state of the catalytic residue His 48, whereas the binding of (R)-oxazolidinone-PC was found to be practically independent of the ionization state of this residue.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Protease II from Moraxella lacunata: cloning, sequencing, and expression of the enzyme gene, and crystallization of the expressed enzyme.

A gene coding for protease II (basic amino acid specific oligoendopeptidase) from Moraxella lacunata was cloned and expressed in Escherichia coli DH1. The transformant harboring a hybrid plasmid, pMPROII-12, with a 3.0-kbp insert at the PvuII-SacI site in pUC19, showed 23-fold higher enzyme activity than M. lacunata. The expressed enzyme from E. coli DH1/pMPROII-12 was purified by 40-80% ammonium sulfate fractionation, chromatography on DEAE-Toyopearl, and Sephadex G-150 gel filtration. The enzyme was most active at pH 6.5 and stable at pH 6.5-9.5. It had an optimum temperature of 35 degrees C for 5 min of reaction and was stable to up to 35 degrees C for 30 min at pH 7.0. Its molecular weight was estimated to be 80,000 by SDS-PAGE and gel-filtration analyses. It enzyme was inhibited by diisopropyl fluorophosphate (DFP) and classified as a serine endoprotease. Its amino acid sequence was 38% homologous to that of the E. coli protease II. By alignment with other members of the prolyl endopeptidase family, the amino acid residues involved in the catalytic triad were deduced to be Ser-534, Asp-619, and His-654. The enzyme was crystallized by the hanging drop vapor diffusion method using PEG 4000 as precipitant.

Amino Acid Sequence↗
PubMed1995

Purification and characterization of the G203T mutant alpha i-2 subunit of GTP-binding protein expressed in baculovirus-infected Sf9 cells.

We expressed the Gly203-->Thr (G203T) mutant of Gi2alpha, which was expected to show a dominant-negative phenotype in Gi2-mediated signal transduction, in baculovirus-inefected Sf9 cells and purified the mutant alpha subunit for its characterization. The rate of dissociation of GDP from G203T Gi2alpha was 3- to 4-fold faster than that from wild type Gi2alpha, but their kappacat values for GTP hydrolysis were almost the same. The affinities of the two Gi2alpha proteins for the beta gamma subunits of G proteins to form alpha beta gamma timers, which served as substrates for pertussis toxin-catalyzed ADP-ribosylation, were the same. In marked contrast, G203T Gi2alpha was unable to form a tight complex with a non- hydrolyzable analog (GTP[gammaS) of GTP; bound GTP[gammaS] was readily released from the mutant Gi2alpha even in the presence of a high concentration of Mg2+. Its susceptibility to tryptic digestion also revealed that GTP[gammaS]-bound G203T Gi2alpha formed a conformation apparently different from that of the GTP[gammaS]-bound form of wild-type Gi2alpha. Both the G203T and wild-type Gi2alpha proteins were capable of coupling with membrane-bound alpha2-adrenergic receptors, resulting in the formation of receptor-G protein complexes with high affinity for agonists. However, GTP[gammaS]-dependent uncoupling from high-affinity receptors was markedly attenuated in the case of G203T Gi2alpha. Thus, G203T-mutated Gi2alpha had a unique property in terms of coupling to membrane receptors, in addition to the previously expected defect in the active conformation of the GTP-bound form of Gi2alpha.

Animals↗
PubMed1995

Intraportal endovascular ultrasonography in the diagnosis of portal vein invasion by pancreatobiliary carcinoma.

OBJECTIVE: The purpose of this study was to determine the value of intraportal endovascular ultrasonography (IPEUS) in the diagnosis of portal vein invasion by pancreatobiliary carcinoma. The authors reported their experiences with this new technique and compared it with conventional imaging technologies, such as portography and computed tomography (CT). SUMMARY BACKGROUND DATA: Pancreatobiliary carcinoma often invades the portal vein. Observation of the echogenic band of the portal vein wall by means of a high-frequency, high-resolution intravascular ultrasound catheter allows for the accurate diagnosis of the portal vein invasion. METHODS: A prospective study of 30 consecutive patients with pancreatobiliary carcinoma (16 pancreatic carcinomas, 8 bile duct carcinomas and 6 gallbladder carcinomas) was performed. In 23 cases IPEUS was performed intraoperatively from the superior mesenteric venous route with an 8 French, 20 MHz intravascular ultrasound catheter. In 7 cases IPEUS was performed before surgery from the percutaneous transhepatic route with a 6 French, 20 MHz intravascular ultrasound catheter. The finding of IPEUS was confirmed by pathologic examination of resected specimens and surgical exploration. The results of IPEUS were compared to those of portography and CT. RESULTS: Intraportal endovascular ultrasonography visualized the portal vein wall as an echogenic band with a thickness of 0.5 mm to 1.0 mm. The diagnostic criterion of portal vein invasion was destruction of this echogenic band. Portal vein invasion was found in 15 of 30 cases. Vascular invasion was confirmed by pathologic examination of resected specimens in 10 patients and operative findings in 5. The sensitivity, specificity, and overall accuracy of IPEUS for diagnosis of portal vein invasion was 100%, 93.3%, and 96.7%, respectively. The values were 80%, 67.7%, and 73.3% for portography and 53.3%, 80%, and 66.7%, respectively, for CT. CONCLUSIONS: Intraportal endovascular ultrasonography provided precise information about the relationship between the pancreatobiliary tumor and the portal vein wall. It was capable of accurately detecting or excluding early invasion of the portal vein wall by pancreatobiliary carcinoma.

Aged↗
PubMed1995

Ifosfamide/carboplatin/etoposide (ICE) for recurrent malignant solid tumors of childhood: a Pediatric Oncology Group Phase I/II study.

PURPOSE: The combination of ifosfamide (I) and etoposide (E) was useful in salvaging patients with recurrent/resistant malignant solid tumors of childhood. Carboplatin (C), active against a number of pediatric cancers, was added to I and E to form a three-drug combination called ICE to improve the response rate. PATIENTS AND METHODS: ICE, consisting of I 1.5 g/m2 plus E 100 mg/m2 i.v.q.d. x 3 plus C i.v. on day 3 only, was given in 21-28-day intervals. C was started at 300 mg/m2, and the dose was escalated in 25% increments, with three evaluable patients treated at each level. RESULTS: Ninety-two patients were enrolled in this phase I/II study between July 1990 and April 1993. A total of 331 courses of ICE was administered. Median courses of ICE received were three (range, 1-16). The maximum tolerated dose (MTD) for C when used in combination was found to be 635 mg/m2. The response rate for ICE at the MTD for C was complete response (CR) 26% and CR + partial response (PR) 53%. The response was even better in those who received C at the MTD: 32% achieving a CR and 63% a CR + PR. Pancytopenia was the dose-limiting toxicity. Thirteen episodes of bacterial infection were reported, none fatal. Only one patient developed a Fanconi-like syndrome. CONCLUSION: The MTD of C when used with I and E was found to be 635 mg/m2. The overall CR + PR rate for all patients treated at all C dose levels was 53%. Best responses were seen in non-Hodgkin's lymphoma, neuroblastoma, soft tissue sarcomas, and Wilms' tumor.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1995

Detection of hepatic micrometastasis in pancreatic adenocarcinoma patients by two-stage polymerase chain reaction/restriction fragment length polymorphism analysis.

Hepatic metastasis and retroperitoneal recurrence generally are considered to be the two primary modes of recurrence in pancreatic cancer. The goal of this study was to determine if patients with pancreatic adenocarcinoma have hepatic and peritoneal micrometastasis at operation. Pancreatic adenocarcinomas are known to have a high incidence of K-ras gene mutations. Liver tissue specimens were obtained from 30 patients (17 with pancreatic adenocarcinoma and 13 with other diseases) with a biopsy needle at operation. Peritoneal washings were obtained during operation from 20 patients with pancreatic adenocarcinoma. Two-stage polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis were used to detect K-ras oncogene mutation at codon 12. Thirteen of 17 pancreatic adenocarcinoma patients had K-ras gene mutations in the liver, whereas all 13 patients with other diseases did not. However, only two of 20 pancreatic adenocarcinoma patients revealed K-ras gene mutation in peritoneal lavage fluids. These results indicate the feasibility of detecting hepatic micrometastasis in patients with pancreatic adenocarcinoma, and imply that PCR/RFLP analysis may be of value in the diagnosis, treatment and follow-up of hepatic metastasis of pancreatic adenocarcinoma.

Adenocarcinoma↗
PubMed1995

Identification of biclonal (duplex) leukaemic cells expressing either CD4+/CD8- or CD4-/CD8+ from a patient with adult T-cell leukaemia/lymphoma.

A 24-year-old Japanese woman was admitted to our hospital in 1987 with a chief complaint of skin eruptions, and was diagnosed as having chronic ATLL. In 1993 the leucocyte count increased gradually to 126.0 x 10(9)/l with 91.5% abnormal lymphocytes expressing two different types of antigenicity, either CD+/CD8- or CD4-/CD8+. Monoclonal integration of human T-cell lymphotropic virus type-I proviral DNA was detected at different sites of the genomic DNA in each cell type. These studies clearly indicate that CD4+/CD8- and CD4-/CD8+ leukaemic cells originated from two independent clones.

Adult↗
PubMed1995

Training in evaluation of expressed emotion using the Japanese version of the Camberwell Family Interview.

To evaluate the effect of training in rating expressed emotion (EE) using the Japanese version of the Camberwell Family Interview (CFI), interrater reliability between a certified rater and a trainee was examined. The material was 65 CFI interviews with the families of 46 schizophrenic patients. The two raters independently rated EE status, critical comments (CC), hostility (H), emotional overinvolvement (EOI), warmth (W) and positive remarks (PR). The interrater reliability was satisfactory with regard to EE status, CC, H, EOI and PR. In the category of W, however, the results were unsatisfactory, as there was no positive development of agreement over time. In fathers, the interrater reliability of W was lower than in other relationships. Use of interview transcripts could contribute to a more satisfactory interrater agreement in EOI, and this is recommended as a useful training tool in EE evaluation.

Adolescent↗
PubMed1995

Intracellular killing of Listeria monocytogenes in the J774.1 macrophage-like cell line and the lipopolysaccharide (LPS)-resistant mutant LPS1916 cell line defective in the generation of reactive oxygen intermediates after LPS treatment.

Listeria monocytogenes is a facultative intracellular pathogen and survives within phagocytic cells by escaping from phagosomes into the cytoplasm. It has been reported that, in vivo, L. monocytogenes is effectively eliminated through cell-mediated immunity, especially by macrophages which have been immunologically activated by cytokines such as gamma interferon (IFN-gamma). However, this killing mechanism for L. monocytogenes and the role of macrophage activation in this bacterial killing are unclear. We demonstrated the listericidal effect of oxidative radicals induced by lipopolysaccharide (LPS) and IFN-gamma, using a macrophage-like cell line, J774.1, and a mutant cell line, LPS1916. LPS1916 cells do not exhibit normal generation of O2- and H2O2 after treatment with 0.1 microgram of LPS per ml, although J774.1 cells generate 100 times the normal level of oxidative radicals with the same LPS treatment. The growth of L. monocytogenes was strongly inhibited in J774.1 cells pretreated with 0.1 microgram of LPS per ml or the combination of 0.1 microgram of LPS per ml and 10 U of IFN-gamma per ml. On the other hand, in LPS1916 cells, the growth of L. monocytogenes was not inhibited by treatment with LPS only, although LPS1916 cells pretreated with the combination of LPS and IFN-gamma showed moderate inhibition of listerial growth. This killing was not influenced by treatment with NG-monomethyl-L-arginine, which is a strong inhibitor of nitrite oxide generation. Interestingly, J774.1 cells treated with LPS did not show enhanced intraphagosomal killing of a nonhemolytic strain of avirulent L. monocytogenes that lacks the ability to escape from phagosomes, and this killing was not influenced by treatment with NG-monomethyl-L-arginine either. These results suggest that the reactive oxygen radicals are more important than nitric oxide in the mechanism underlying the intracellular killing of virulent L. monocytogenes and that there seem to be different killing mechanisms for virulent and avirulent strains of L. monocytogenes in activated-macrophage cell lines.

Animals↗
PubMed1995

Association of malachite green-positive material with heparan sulfate proteoglycan double tracks in basement membrane of mouse kidney tubules.

The presence of lipids in the basement membrane of the mouse kidney tubules was examined by histochemical staining with malachite green. Pieces of mouse kidney cortex were immersed in a fixative containing 3% glutaraldehyde and 0.1% malachite green in 0.067 M sodium cacodylate buffer, pH 6.8, for 18 hr at 4 degrees C. Control tissue was fixed in the same way except that no malachite green was added to the fixative. The tissue pieces were cryoprotected, frozen in Freon 22, and subjected to freeze-substitution in dry acetone containing 1% OsO4. Thin sections of Epon-embedded specimens were observed by electron microscopy at first without uranyl-lead counterstaining. The basement membrane of mouse kidney tubules was positively stained in a pattern composed of an irregular assembly of 5-8-nm wide strands. The nature of these malachite green-positive strands was further examined by counterstaining thin sections with uranyl-lead, and they were identified as 4.5-5-nm wide ribbon-like "double tracks" previously characterized as the form taken by heparan sulfate proteoglycan in basement membranes. It is concluded that lipids are present in the basement membrane of mouse kidney tubules in association with heparan sulfate proteoglycan.

Animals↗