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Volume 42, Issue 2
Displaying 1-13 of 13 articles from this issue
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Kiyoshi Takatsuki1998 Volume 42 Issue 2 Pages 73-77
Published: June 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSRetroviruses contain an RNA-dependent DNA polymerase (a reverse transcriptase) that directs the synthesis of a DNA form of the viral genome after infection of a host cell. HTLV-1 and HIV-1 are known pathogenic human retroviruses which cause adult T-cell leukemia/lymphoma (ATL) and AIDS, respectively. ATL was originally reported in Japan in 1976. AIDS was first recognized in the United States in 1981 and the global pandemic of AIDS became a menace to the mankind. The purpose of this lecture is to present the current information available on the epidemiology, pathogenesis, symptomatology, diagnosis, treatment and prevention of these two diseases.View full abstractDownload PDF (594K) -
Akira Shimizu1998 Volume 42 Issue 2 Pages 79-85
Published: June 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSDirect examination of hemolysate by electrospray ionization mass spectrometry may lead to the rapid ascertainment of variant hemoglobins (Hb) and to the quantification of glycated Hb. Plasma and cell proteins other than Hb can be prepared by immunoprecipitation prior to MS. The detected variants were sequenced by MS using enzyme cleaved peptides. We detected various variants of Hb, transthyretin, and CuZn-superoxide dismutase. New variants included; 1) Hb Sagami [β139 (H17) Asn→Thr], and 2) Hb Hokusetsu [β52 (D3) Asp→Gly]. These were studied to clarify the cause of unusual profiles of HPLC for HbAlc measurement. Hb Sagami was an electrophoretically silent and isopropanol test-negative variant, detected solely by ESIMS. Other new variants included; 3) a variant transthyretin, amyloidogenic, [38 Asp→Ala], and 4) another variant transthyretin, non-amyloidogenic, [101 Gly→Ser]. Variant transthyretins are usually difficult to detect by electrophoresis. Samples containing variant Hb or high carbamylated Hb gave erroneous results of glycated Hb by routine HPLC and immunoassay, but did not by MS.View full abstractDownload PDF (2460K) -
Kaoru Miyazaki, Naohiko Koshikawa1998 Volume 42 Issue 2 Pages 87-92
Published: June 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSTumor cells produce various types of matrix-degrading proteinases which play essential roles in tumor cell invasion through basement membranes and connective tissues. Highly sensitive analysis of proteinases by zymography on gelatin- or casein-containing gels has shown that tumor cells frequently secrete matrix metalloproteinases (MMPs) including gelatinases A and B, interstitial collagenase, stromelysin and matrilysin, as well as the serine proteinase trypsin. On the other hand, analysis by reverse zymography has revealed that tumor cells also secrete various kinds of inhibitors for MMPs and trypsin. These results suggest that extracellular proteolysis has been regulated by a balance between various kinds of proteinases and their inhibitors. The zymography and reverse zymography can be widely applied to studies on proteinases present in various biological fluids and tissues.View full abstractDownload PDF (4039K) -
Gel shift assayTakumi Kawabe, Toshifumi Tetsuka, Takashi Okamoto1998 Volume 42 Issue 2 Pages 93-96
Published: June 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSDNA binding proteins play important role in the DNA replication, recombination, transcription and the viral integration to genome. Several methods to analyse these DNA binding proteins such as methylation interference and DNase protection analysis were developed. Among them gel shift analysis has been widely used because of its simplicity, high sensitivity and rapidity. We review the detail of this method by refering to the example of the analysis of the redox regulation of the transcription factor NF-κ B binding to DNA. In this analysis the DNA binding activity of the NF-κB was abolished by the treatment with the oxidizing reagent diamide. Thioredoxin (ADF), but not glutathione efficiently activated NF-κB binding activity to its consensus DNA sequence.View full abstractDownload PDF (4066K) -
Toshifumi Tetsuka, Takumi Kawabe, Takashi Okamoto1998 Volume 42 Issue 2 Pages 97-101
Published: June 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSThe phosphorylation and/or dephsophorylation of transcription factors modulate their nuclear translocation and/or transcriptional activities. The in-gel kinase assay was used to identify the kinases which phosphorylate transcription factors, c-Jun and NF-κB. Recombinant c-Jun or NF-κB was copolymerized in an SDS-PAGE gel as a substrate. After electrophoresis, the gel was denatured in 6M guanidine and then renatured in a buffer containing 0.04% Tween 40. The renatured substrates and kinases in the gel were incubated with [γ-32P] ATP. The renatured kinases successfully phosphorylated transcription factors. The in-gel kinase assay revealed that interleukin-1 stimulates c-Jun aminoterminal kinase (JNK). This method also showed that c-AMP dependent protein kinase (PKA) phosphorylates the p 65 subunit of NF-κB. Thus the in-gel kinase assay is a useful method to identify the kinases which phosphorylate transcription factors.View full abstractDownload PDF (2719K) -
Toshikazu Hada1998 Volume 42 Issue 2 Pages 103-108
Published: June 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSFour distinct types of human alkaline phosphatase (AP) have been identified in healthy individuals. These APs are of liver/bone/kidney, adult intestine, placenta and germ cell. An AP was found in sera, ascitic fluid or cancer tissues of patients with hepato-cellular carcinoma which had a different electrophoretic mobility from those of normal enzymes on 5% PAGE. This enzyme initially regarded as a variant of placental AP in view of its enzymatic and immunological properties. Afterwards, on the basis of the identity of this enzyme with one of the two AP isoenzymes produced in FL-amnion cells, this enzyme was designated as the Kasahara isoenzyme (K. I.) after the first patient. Gene analysis of K. I. revealed that the amino acid sequence of K. I. is identical with that of adult intestinal AP, suggesting that the different electrophoretic mobility of K. I. may be due to the aberrant glycosylation of adult intestinal AP. Clinicopathological findings are also reported.View full abstractDownload PDF (2041K) -
Yasushi Takagi1998 Volume 42 Issue 2 Pages 109-112
Published: June 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSCreatine kinase (CK, EC 2.7.3.2) has a important role in muscle contraction and is rich in muscle tissue. The CK has three isoenzymes, designated as CK-MM (also termed CK3), CK-MB (CK2) and CK-BB (CK1). These isoenzymes are clearly separated by an electrophoresis. Myocardium contains significant quantities of CK-MB. Demonstration of elevated levels of CK-MB is considered the most specific indicator of acute myocardial infarction (AMI). CK-MM is resolved into three isoforms (MM1, MM2, MM3) by prolonged electrophoresis. The CK-MM3 in myocardium is released into blood and changes to CK-MM2, then CK-MM1 post-translationally. The detection of an elevated percentage of CK-MM3 in serum of patients with AMI indicates only a relatively recent myocardial injury like a myoglobin. In recent years, the unusual CK isoenzyme bands displaying electrophoretic properties that differ from the major isoenzyme fractions were observed. These atypical forms are generally of two types and are referred to as CK-linked immunoglobulin and mitochondrial CK (CK-m). The CK-m is detected in serum of extensive tissue damage, causing breakdown of the mitochondria.View full abstractDownload PDF (3969K) -
Yoshiaki Katayama, Koji Yoneda1998 Volume 42 Issue 2 Pages 113-116
Published: June 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSThe change of mitochondria aspartate aminotransferase (mAST)/soluble AST (sAST) ratio was examined in 22 cases of acute myocardial infarction (AMI). The mAST/sAST was 0.408±0.189 at admission to a hospital (2.9±1.6h). The mAST/sAST decreased to normal value rapidly after peak and then increased again gradually. The decrease ratio of mAST/sAST per minute at early stage of 8 cases, who were succeeded to reperfusion, was 0.28±0.20, and that was significantly higher than of conventionally treated 7 cases and non-reperfused 7 cases (0.11±0.07). These results indicated that (1) mAST/sAST may be an excellent indicator for AMI in early stage. (2) The decrease ratio of mAST/sAST would predict whether reperfusion is successful or not at an earlier stage of AMI.View full abstractDownload PDF (323K) -
Taizo Hayashi1998 Volume 42 Issue 2 Pages 117-122
Published: June 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSSerum PGAM activities and the percentage of brain-subunit activity against total activity (B/T) were determined in the cases of apoplexy and traumatic injury on head. PGAM activities already increased and peaked whthin 2h after the stroke. This findings were also recognized in the cases with head-trauma. They gradually decreased in 12h, 24h and 3 day after the attack. The grade and duration on the elevation of PGAM activities were more dominant in the serious cases than in the slight cases. In these cases with apoplexy and head-traum, B/T ratio almost remained in the normal range, namely about 90%. In the CSF, PGAM activities were consist of B-subunit, and significantly correlated with LD-activities and the protein content. In the experiment on SHRSP, serum PGAM activities were markedly increased during the stroke, compared with that of control rat. We suggest that serum PGAM activities would be an useful marker of apoplexy and the other brain damage.View full abstractDownload PDF (704K) -
Nobue Sakai, Kiyoko Shiba, Nobuo Hiratsuka, Seijin Hosaki, Kenzi Nishi ...1998 Volume 42 Issue 2 Pages 123-129
Published: June 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSWe established a method for quantitative analysis of urinary components by capillary electrophoresis using SDS containing buffer. This method was applied for analyses of urine samples from healthy subjects, diabetic patients at pre- and early nephropathic stages. The urine electropherograms in healthy subjects, diabetics at pre- and early nephropathic stages had 44, 46 and 53 peaks at maximum, respectively. Out of these peaks, 10 peaks were identified. The urine components were separated by this method according to their molecular sizes, which were estimated from their migration times using standard proteins. However, the glycoproteins linked with a larger number of sugar chains, such as α1-acid glycoprotein and Tamm-Horsfall protein showed a tendency to be overestimated in their molecular sizes. Generally, the urine electropherograms of healthy subjects and diabetics revealed that the peak numbers increased especially in higher molecular size in accordance with development of renal impairment. From these results, the urine electropherograms give us a suitable information not only as the stage of glomerular but tubular impairment and an early prediction of diabetic nephropathy.View full abstractDownload PDF (692K) -
Takanori Moriyama, Tatsuo Tozawa, Satoshi Hirata, Hisami Ikeda1998 Volume 42 Issue 2 Pages 131-136
Published: June 15, 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSElectrophoretic and chromatographic characterization of macroamylasemia induced by hydroxyethyl starch (HES) was performed. HES-induced macroamylasemic sera were obtained from the patients subjected to intravenous infusion of HES solution during surgery. Macromolecular amylase was observed in the patient's sera obtained from termination of the infusion of HES to 24h post-operation by gel-permeation high-performance liquid chromatography (HPLC) analysis. However, anomalous bands were detected in the patient's sera only immediately or two hours after termination of the infusion by isoamylase electrophoretic analysis. In the samples of non-macroamylasemic serum mixed with HES solution in vitro, the anomalous pattern was observed only in the sample with volume ratio of 1:1 and/or 1:1/2. In contrast, the peak of macromolecular amylase was detected in the all samples with the ratio from 1:1/2 to 1:1/20. From the results, it was recognized that gel-permeation HPLC analysis was a more sensitive method in HES-induced macroamylasemia than isoamylase electrophoretic analysis.View full abstractDownload PDF (3206K) -
1998 Volume 42 Issue 2 Pages 139a
Published: 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSDownload PDF (84K) -
1998 Volume 42 Issue 2 Pages 139b
Published: 1998
Released on J-STAGE: March 31, 2009
JOURNAL FREE ACCESSDownload PDF (84K)
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