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Volume 42, Issue 4
Displaying 1-11 of 11 articles from this issue
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Kohzoh Imai1998 Volume 42 Issue 4 Pages 217-223
Published: December 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSGenetic analysis for tumor cells has recently been advanced due to the development of the new biotechnology. In this review article, recent advancement of the genetic instability in digestive tract cancers and hypermethylation of the promoter region of the gene was reviewed. The new data from our laboratory concerning these points was presented in addition to the detailed description of the methodology.View full abstractDownload PDF (2350K) -
Eiji Miyoshi, Naoyuki Taniguchi1998 Volume 42 Issue 4 Pages 225-231
Published: December 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSα1-6 Fucosyltransferase (α1-6 FucT) catalyzes the transfer of fucose to innermost GlcNAc in N-glycans. This structure on α-fetoprotein has been clinically used for early diagnosis of hepatocellular carcinoma. In this study, expression of α1-6 FucT was investigated in rat and human hepatocarcinogenesis and biological significance of α1-6 FucT on hepatoma was investigated in terms of metastasis. While α1-6 FucT was scarcely detected in normal liver, it was enhanced in LEC rat hepatoma, but not in their surrounding tissues. In contrast, high expression of α1-6 FucT mRNA was observed in human liver cirrhosis tissues as well as in hepatoma tissues. We established human hepatoma cell lines which express high levels of α1-6 FucT by transfection of α1-6 FucT gene and investigated intrahepatic metastasis after splenic injection to athymic mice. Tumor formation in the liver was dramatically suppressed in α1-6 FucT transfectants. While there were no differences of cell invasiveness and cytotoxicity to lymphocytes, cell adhesion to nonparenchymal liver cells in culture was significantly inhibited in α1-6 FucT transfectants as compared to those of controls. Attachment of α1-6 FucT transfectants to a fibronectin or laminin coated dish was decreased compared to controls. Two-dimensional electrophoresis followed by lectin blot showed certain glycoproteins were α1-6 fucosylated and might be linked to suppression of intrahepatic metastasis. Collectively, although α1-6 FucT was enhanced in rat and human hepatocarcinogenesis, overexpression of α1-6 FucT suppressed intrahepatic metastasis of hepatoma.View full abstractDownload PDF (5980K) -
Susumu Saito1998 Volume 42 Issue 4 Pages 233-237
Published: December 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSAllelic deletions of chromosome 6q that occur frequently in ovarian cancers imply the presence of a putative tumor suppressor gene in this chromosomal vicinity. We analyzed DNA from 32 patients with ovarian carcinomas for loss of heterozygosity at loci on the distal portion of chromosome 6q and constructed a detailed deletion map. The map indicated a commonly deleted region between loci D 6S 149 (defined by CI6-24) and A 2, which are estimated to be 300kb apart on the basis of our cosmid contig map. DNA sequencing in the region disclosed the presence of AF-6, a gene that had been identified as the ALL-1 fusion partner involved in acute myeloid leukemias with t (6; 11) (q27; q23) translocations. Subsequent screening of the AF-6 gene in ovarian carcinomas revealed no mutations.View full abstractDownload PDF (538K) -
Takahiro Inoue, Kiyoshi Ohkawa1998 Volume 42 Issue 4 Pages 239-243
Published: December 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSWe have investigated the oligosaccharide structures of serum transferrin (Tf), and of serum carbohydrate-deficient transferrin (CDT) in particular, from patients with alcoholic liver disease (ALD) using lectin affinity electrophoresis coupled with antibody-affinity blotting and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by western blotting. By concanavalin A-affinity electrophoresis, serum CDT was separated mainly into weakly reactive and non-reactive Tfs which showed slower electrophoretic mobilities than those from the healthy controls. Moreover, nearly all of the serum CDT was non-reactive with datura stramonium agglutinin. On SDS-PAGE, the molecular masses of serum CDT were estimated to be approximately 75 and 72kDa, which corresponded to those of partially and completely deglycosylated Tf from the healthy controls (78kDa), respectively. These results indicated that the oligosaccharide structures of serum CDT from patients with ALD show not merely a loss of terminal sialic acids, but also the absence of asparagine-N-linked oligosaccharides. Similar result was found with serum α1-acid glycoprotein.View full abstractDownload PDF (3697K) -
Takanori Nakajima, Shin-ichiroh Kawai, Toshiyuki Matsunaga, Shigeru Ho ...1998 Volume 42 Issue 4 Pages 245-249
Published: December 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSWe obtained a monoclonal antibody 9F 5-3 a against oxidative high-density lipoprotein (HDL) modified with CuSO4. In addition, we have examined the epitope of 9F 5-3 a and the localization of oxidized HDL (oxHDL) in atheromatous plaque. The 9F 5-3 a was reacted strongly with oxHDL and to a lesser degree with oxidized low-density lipoprotein (oxLDL) and native LDL. No reactivity was found with native HDL. Furthermore, the cross-linkage of apolipoprotein A-I and/or apolipoprotein A-II was accelerated by HDL oxidation, and the crosslinked apolipoprotein forms (32, 60, 90, 120 and 200-kDas) were selectively recognized by the 9F 5-3 a antibody. OxHDL particles reacting with the 9F 5-3 a antibody were localized histochemically in the luminal side of intima of atheromatous plaques in human abdominal aortae. Using a ligand receptor assay for oxHDL, a major 130-kDa band was detected in cultured human aortic endothelial cells. These results suggested that oxHDL particles and its putative binding protein were present in atheromatous plaques and endothelial cells, respectively.View full abstractDownload PDF (5441K) -
Fumio Itoh, Yuji Hinoda1998 Volume 42 Issue 4 Pages 251-255
Published: December 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSSchirrous gastric cancer is one of the most mortal neoplasms and is characterized by histopathological features of poorly differentiated type and infiltrative growth. However, very little is known about the genetic mechanisms of this malignant tumor, which is difficult to analyze due to the proliferation of scirrhous stroma. The aim of this study was to apply an improved representational difference analysis (RDA) in order to identify the genetic alterations in schirrous gastric cancer. To monitor the efficiency of RDA, mouse DNA fragment was added to a tester prepared from the genomic DNA, at the rate of a single copy per haploid genome (Efficiency-monitored RDA (EM-RDA)). The difference products were analyzed by Southern blot with the mouse DNA as a probe. The real difference products, other than the mouse DNA, were subcloned. As a result, added control DNA fragments were sufficiently amplified through 3 rounds of subtraction and the high efficiency of the subtraction was confirmed. In summary, efficiency-monitored RDA was demonstrated to raise the reliability of RDA. Furthermore this method successfully contributed to the detection of fragment in the schirrous gastric cancer.View full abstractDownload PDF (3375K) -
Shugo Nawata, Hiroshi Kato, Kazuyuki Nakamura1998 Volume 42 Issue 4 Pages 257-263
Published: December 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSRecently, tumor markers, which have been developed for the diagnosis and evaluation for cancer, are regarded as new key tools to study the malignant behavior of cancer, and therefore, much attension has been focused on their biological functions. Squamous cell carcinoma (SCC) antigen is a tumor marker of SCC arising in various sites. We have investigated the biological function and heterogeneity of this tumor marker by electrophoretic methods. In this paper, we report that SCC antigen belongs to the inhibitory type of serine protease inhibitor family, indicating that SCC antigen may affect malignant behavior such as tumor invasion. We also describe the different pattern of SCC antigen by two-dimensional electrophoresis in normal and cancer tissues, and indicate that the heterogeneity of SCC antigen results mainly from the presence of two homologous genes, SCCA 1 and SCCA 2 gene.View full abstractDownload PDF (5743K) -
Corinne E. M. Olesen, Jennifer Mosier, Chris S. Martin, John C. Voyta, ...1998 Volume 42 Issue 4 Pages 265-279
Published: December 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSThe use of 1, 2-dioxetane chemiluminescent enzyme substrates, including AMPPD®, CSPD®, CDP® and CDP-Star® for alkaline phosphatase and Galacton-Star® substrate for β-galactosidase, provides highly sensitive detection for numerous immunoassay and nucleic acid detection formats. Enzyme cleavage of the 1, 2-dioxetane substrate generates a metastable anion intermediate that decomposes with the concomitant emission of light. Light emission exhibits glow kinetics, enabling the use of multiple imaging platforms for signal detection, including film-based, luminometers, low-light sensitive camera and phosphor screen instrumentation systems. Applications include both membrane-based immunodetection of proteins and nucleic acid blot hybridization, and solution-based immunoassays and nucleic acid capture/hybridization assays performed in a microwell plate.View full abstractDownload PDF (4446K) -
Yoshihiro Ashihara, Mitsuo Isomura1998 Volume 42 Issue 4 Pages 281-285
Published: December 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSChemiluminescent immunoassays have commonly been used in clinical laboratories because of their high sensitivity and wide dynamic range. We employed highly sensitive and stable 1, 2-dioxetane substrates, AMPPDTM and CSPDTM for alkaline phosphatase (ALP) and applied chemiluminescent enzyme immunoassay (CLEIA) for the detection of tumor markers and antibodies to infectious viruses. Chemiluminescent enzyme immunoassay using AMPPD as ALP substrate showed S/N values of more than 80 times those of CLEIA using fluorescent or colorimetric substrate. The chemiluminescent reaction using CSPD reached a plateau at 5 minutes. We observed a higher chemiluminescent signal by energy transfer using several fluorescent reagents in the presence of AMPPD and the enhancer BDMQ. Gelatin-coated ferrite particles as a solid phase showed a high dispersion floating property and very low background noise. We established an extremely sensitive CLEIA for IL-6 in which the detection limit was 0.1pg/ml (zero+2 SD). We also applied gelatin solid phase to establish a sensitive CLEIA by reducing background noise through the release of an immne complex consisting of solid antibody, antigen and labeled antibody from ferrite particles using gelatinase. In this method, 7.5 times higher S/N than conventional CLEIA was obtained.View full abstractDownload PDF (590K) -
Katsuo Kubono, Ikunosuke Sakurabayashi1998 Volume 42 Issue 4 Pages 287-293
Published: December 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSIn order to standardize the lipoprotein(a) [Lp(a)] measurement, the International Federation of Clinical Chemistry Working Group on Lp(a) (IFCC Lp(a) WG) and the Japan Society of Clinical Pathology Working Group on Lp(a) Standardization were established in 1995, 1994, respectively. In the activities on IFCC Lp(a) WG, eight proposed reference materials (PRM) were compared for their analytical performance, commutable properties, method harmonization, and selected PRM-2B (Behring) as the most suitable PRM. In the next phase of the IFCC Lp(a) WG, a value transfer protocol will be used to transfer a Lp(a) value from the PRM-2 for Lp(a) to manufactures' master calibrators, and WG will submit the results to WHO. On the other hand, it is recommended that the Lp(a) protein concentration should be expressed in molar units for more accurate Lp(a) measurement. So it will be expected that new clinical estimation is investigated.View full abstractDownload PDF (7577K) -
Zensuke Ogawa, Hajime Itoh1998 Volume 42 Issue 4 Pages 295-298
Published: December 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSOn the enzyme committee of Japan Society of Clinical Chemistry, the studies of enzyme assay for the recommendation methods was started from 1975. Already, recommendation methods of aspartate aminotransferase, alanine aminotransferase, creatine kinase, alkaline phosphatase, lactate dehydrogenase and γ-glutamyltransferase were published. And new, two kinds of enzymes, amylase and cholinesterase were in progress to discuss for the consensus.View full abstractDownload PDF (458K)
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