Biomedical subjects

M Kanai

Publications and source records attributed to M Kanai.

At least 19 recordsLinked to original sources

PubMed1992

Identification and characterization of apolipoprotein(AII-E2-AII) complex in human plasma lipoprotein.

A new apolipoprotein complex designated as the apo(AII-E2-AII) complex was identified in the lipoprotein fractions of human plasma with apoE phenotypes containing apoE2 (E4/E2, E3/E2, and E2/E2). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by an immunoblotting assay using anti-apoE or anti-apoAII antibodies, established that the apo(AII-E2-AII) complex, with a molecular weight of 58,000, was identical to the complex consisting of apoE and apoAII, and that it also dissociated following reduction with beta-mercaptoethanol. This new complex was also demonstrated to be distinct from the apo(E-AII) complex and apoE monomer by isoelectric focusing, in the samples that were not treated with beta-mercaptoethanol. In apoE phenotype E3/E2, the apo(AII-E2-AII) complex was primarily included in the high-density lipoprotein (HDL, 1.063 < d < 1.21 g/ml) fraction, but was also observed in a small quantity in the very-low-density lipoprotein (VLDL, d < 1.006 g/ml) fraction. For further characterization, the apo(AII-E2-AII) complex was isolated by preparative SDS-PAGE, and no contamination of apo(E-AII) complex and apoE monomer was detected by immunoblotting assay using an anti-apoE antibody. It was confirmed by an enzyme-linked immunosorbent assay (ELISA) system that a molecular ratio between apoAII monomer and apoE in the isolated apo(AII-E2-AII) complex was approx. 2, when the apo(E-AII) complex was used as a standard with the ratio of 1:1. It indicates that the apo(AII-E2-AII) complex is formed from two molecules of apoAII monomer and one molecule of apoE.(ABSTRACT TRUNCATED AT 250 WORDS)

Apolipoprotein A-II↗

PubMed1992

Identification of AJ-2615 and its S-oxidized metabolites in rat plasma by use of tandem-mass spectrometry.

The structural elucidation of the metabolites of a new calcium entry blocker, AJ-2615 (AJ), in rat plasma is described. Metabolites in a crude plasma extract from spontaneously hypertensive rats were identified without chromatographic separation by fast atom bombardment tandem mass spectrometry. When the plasma extract was examined by using parent ion scans, the presence of the oxidized metabolites of AJ was suggested. These metabolites were identified as the S-oxide and the S,S-dioxide by comparing their daughter ion spectra with those of authentic samples. The presence of the two diastereomeric S-oxides of AJ in the plasma extract was ascertained by high-performance liquid chromatography. Their relative configurations were determined by infrared and proton nuclear magnetic resonance spectra.

Animals↗

PubMed1992

Immunohistochemical demonstration of glycoconjugates bearing the type 2 chain-backbone structure in human fetal, normal and neoplastic gastrointestinal tract.

Immunohistochemical distributions of carbohydrate antigens based on the type 2 chain in normal as well as fetal and neoplastic tissues of human gastrointestinal tract were investigated with a monoclonal antibody (MAb) H11 (specific for type 2 chain) alone and in combination with the two MAbs MSG15 (for alpha 2----6 sialylated type 2 chain) and IB9 (for the alpha 2----6 sialylated type 2 chain and glycoproteins having NeuAc alpha 2----6Gal-NAc), and 188C1 (for short- and long-chain Lex antigens) and FH2 (for the long-chain Lex antigen). In the pyloric mucosa of secretors, the type 2 chain is oncodevelopmentally expressed, but in non-secretors it is detected in surface mucous cells of normal gastric mucosa. The alpha 2----6 sialylation, which is confined to endocrine cells of normal pyloric mucosa, occurs in fetal and carcinoma tissues. Irrespective of the secretor status, the short- and the long-chain Lex antigens can be detected in mature and immature glandular mucous cells of normal gastric mucosa, respectively; both antigens are also expressed in fetal and carcinoma tissues. In the colon, the type 2 chain and its alpha 2----6 sialylated counterpart are expressed in an oncodevelopmental manner. The short- and the long-chain Lex antigens are significantly enhanced in colonic carcinoma. The glycoproteins with NeuAc alpha 2----6GalNAc residues appear in gastric and colonic carcinoma as well as intestinalized gastric mucosa and transitional mucosa. Thus, some of these antigens were distinctively expressed in certain epithelial cells lining the normal gastrointestinal tract depending on maturation and patients' secretor status, and some were oncodevelopmental or carcinoma-associated antigens of the human gastrointestinal tract.

Antibodies, Monoclonal↗

PubMed1992

Distribution of retinol-binding protein in the human digestive tract.

By employing polyclonal antibodies for retinol-binding protein (RBP), its distribution in the human pancreas and digestive tract mucosa was compared with those of transthyretin (TTR) and various peptide hormones. The materials used included surgically removed pancreas, esophagus, stomach, small and large intestines. Paraffin sections were stained by the indirect immunoenzyme method. The results indicate that RBP-containing cells are found in the pancreas and the gastrointestinal mucosa, but most frequently in the gastric antrum and duodenum. In the pancreas, RBP-containing cells are found in the islets and among acinar and ductal epithelial cells, and consistently stain for chromogranin A. RBP-containing cells in the gastrointestinal mucosa showed typical features of endocrine cells and also stained for chromogranin A. The distribution of TTR in these tissue sites resembled that of RBP, but the immunoreactive intensities of both peptides altered independently. Comparison of the distribution of RBP, TTR, and various gastrointestinal peptide hormones revealed that the distribution of RBP coincided with none of the other peptides, although some of the RBP-containing cells stained for most of the peptides examined and vice versa. These results suggest that RBP may be a consistent component of gastrointestinal endocrine cells.

Antibodies↗

PubMed1992

Clinical evaluation of pancreatic cancer-associated mucin expressing CA19-9, CA50, Span-1, sialyl SSEA-1, and Dupan-2.

We have previously described the purification and partial characterization of a new pancreatic cancer-associated antigen, a pancreatic cancer-associated mucin expressing CA19-9, CA50, Span-1, sialyl SSEA-1, and Dupan-2. This study describes the clinical evaluation of various assay systems for this antigen which depend on measuring respective serum levels. Elevated levels of antigen were detected in the sera from both patients with malignant and non-malignant diseases. However, elevated serum levels of CA19-9 and Lewisa and Lewisb epitopes on moieties were restricted to pancreatic and biliary tract cancers, although adequate sensitivity was not attained. Coordinate evaluation of these three markers improved the sensitivity to some extent without loss of specificity for the diagnosis of pancreatic and biliary tract cancers, because of the heterogeneity of the coexpression of these epitopes. We developed additional assay systems with a combination of this antigen and two lectins (Bauhinia purpurea (BPA) and Vicia villosa (VVA)). Elevated levels of BPA- and VVA-reactive antigens were detected in 41% and 31%, respectively, of pancreatic cancer sera samples. Few patients with chronic pancreatitis had an elevated serum level of either antigen, and higher elevated levels of these markers were restricted to the sera of patients with malignancies. Our results suggest that this antigen is found in the sera of patients with various conditions and in the sera of normal subjects but that antigens bearing CA19-9 or Lewisa or Lewisb epitopes and an altered carbohydrate structure recognized by BPA and VVA lectins are preferentially present in the sera of patients with pancreatic and other malignancies.

Antigens, Neoplasm↗

PubMed1992

[Studies on the fragments of FDP in 4 patients with DIC].

We previously studied fibrinolysis and fibrinogenolysis by analyzing fragments of fibrin/fibrinogen degradation products (FDP) employing sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. In this report, we characterized the fragments of FDP in four patients with disseminated intravascular coagulation (DIC), that were caused by various diseases. In the patients suffering from acute lymphoblastic leukemia (case 1) and acute suppurative cholangitis (case 3), DD and DY/X fragments resulting from fibrinolysis accounted for the most part of the FDP fragments. In case 3, D fragments resulting from fibrinogenolysis were also observed to much less extent. In a DIC associated with acute myeloblastic leukemia (case 2), both fibrinolysis and fibrinogenolysis were increased and resulted in high levels of D, Y and DY/X fragments, concomitant with moderate levels of DD and high molecular weight (HMW) fragments in the patient's sera. The increased fibrinogenolysis in this case was attributed to accelerated activation of plasmin. In a DIC patient of case 4, who underwent an operation due to hepatocellular carcinoma, marked increase in DY/X and HMW fragments and slight increase in DD fragment were observed on the day of operation. Hyperfibrinolysis documented in case 4 was explained by both increased production of thrombin and moderately accelerated activation of plasmin. Both qualitative and quantitative changes in the fragments of FDP during the courses of treatment in two cases of DIC were also noted. In summary, each underlying disease expresses characteristic pattern of FDP fragments in DIC.

Adolescent↗

PubMed1992

Mitochondrial dysfunction in the non-obstructed lobe of rat liver after selective biliary obstruction.

In order to elucidate the function of non-obstructed hepatic lobe during partial cholestasis, we have examined the effects of selective biliary obstruction on the mitochondrial energy transducing system in rat liver. The non-ligated lobe became hypertrophic after ligation of the bile ducts that drained 90% of the liver, and there was no increase in the level of serum bilirubin, alkaline phosphatase activity, or bile acids. However, mitochondrial oxidative phosphorylation function, specific enzymic activities of the oxidative phosphorylation complexes, and the subunit contents of these complexes were markedly decreased in the non-obstructed lobe at 4 weeks after the 90% biliary obstruction. There was no increase in the specific content of mitochondrial DNA. The mitochondrial energy transducing system in the non-obstructed lobes is not enhanced, but is significantly impaired during prolonged selective biliary obstruction, despite normal serum biochemical data and hypertrophy of the non-ligated lobe. These results imply early release of obstruction of cholestatic lobe, e.g. by biliary drainage, would be beneficial for maintaining the mitochondrial function in the non-cholestatic lobe.

Animals↗

PubMed1992

[Differentiation of fibrinolysis and fibrinogenolysis by analysis of FDP fragments].

We previously analysed the fragments of fibrin/fibrinogen degradation products (FDP) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) combined with immunoblotting. In this report, we studied the semi-quantitative analysis of fibrinolysis (degradation of cross-linked fibrin) and fibrinogenolysis (degradation of fibrinogen and/or unstable fibrin) of patients' samples by our method. In vitro study of FDP made it clear that an appearance of D fragment confirmed fibrinogenolysis and an appearance of DD fragment and/or high molecular weight fragments which have higher molecular weight than DY or X fragment confirmed fibrinolysis. In addition, a study with mixtures of various concentrations of fibrin degradation products (FbDP) and fibrinogen degradation products (FgDP) demonstrated a dose dependent intensity of band by immunoblot method. These results show that our method is favorable for the semi-quantitative analysis of fibrinolysis and fibrinogenolysis. We applied the method to 6 samples from patients with disseminated intravascular coagulation (DIC). Consequently, fibrinogenolysis was observed in all of 6 samples, in which fibrinogenolysis was more enhanced than fibrinolysis in one sample, and an equivalent degree of fibrinolysis and fibrinogenolysis were observed in 3 of 6 samples. Although our method was probably devoid of the ability to distinguish FgDP from degradation products of unstable fibrin, these findings indicate that fibrinogenolysis is, at any rate, enhanced in the majority of patients with DIC, besides fibrinolysis.

Electrophoresis, Polyacrylamide Gel↗

PubMed1991

Simple and accurate determination of methylpyrazines in biofluids using high-performance liquid chromatography.

The determination of six methylpyrazines was performed using high-performance liquid chromatography (HPLC). Methylpyrazines were simultaneously extracted and injected onto a silica gel column with a syringe-type minicolumn packed with diatomaceous earth granules. The extraction-injection solvent used was dichloromethane and the mobile phase solvent for HPLC was dichloromethane containing 0.08% of 1.65 M ammonia solution and 0.5% of methanol. Methylpyrazines were detected using an ultraviolet detector set at 275 nm. Linear relationships between the amount of sample and peak height were confirmed from 50 ng/ml to 10 micrograms/ml of the biofluids. When an aliquot of 10 microliters of biofluid was introduced to the minicolumn, the detection limit of methylpyrazines was as low as 30 ng/ml with each pyrazine derivative. The method is simple and accurate and is thus applicable to pharmacokinetic studies which are performed on animals. The results showed that the possible pharmacological effects of methylpyrazines might be evaluated pharmacokinetically using this newly developed technique.

Animals↗

PubMed1991

[Simple process of management of central venous catheter].

In our institution, central venous catheterization is a routine procedure in terms of keeping the veins in terminal cancer patients and postoperative patients, as well as on purpose to perform high calorie transfusion. Here we describe our efforts to simplify the process. The process is characterized by the intermittent transfusion with filling up of 100-fold diluted heparin; no use of filter at the usage; but free side infusion from a trigonal active plug, which is also used for blood transfusion. These patients are permitted to bathe and stay out. During a recent 2 years, a total of 111 patients was managed with this process. Duration of catheterization ranged from the shortest of 5 days to longest of 96, with a mean of 27.5 and total days of 3058. Catheter-induced fever as a complication was observed in only 3 cases (2.7%). This procedure has a considerably simplified process, which can be said to depart from the conventional ones in terms of keeping clean, however, in actual clinical routine, it is extremely safe and valuable.

Catheterization, Central Venous↗

PubMed1991

Hepatocyte growth inhibitory factor derived from HTLV-I(+) T cell lines: effect on the epidermal growth factor-dependent proliferation of rat hepatocytes.

A human T cell leukemia virus-I infected T cell line, ATL-2, produces an interleukin-2 receptor inducing factor, adult T cell leukemia (ATL)-derived factor (ADF). In the conditioned medium (CM) of ATL-2, we found an inhibitory activity on the epidermal growth factor (EGF)-dependent proliferation of primary cultured rat hepatocytes, measured by cell number and [3H]thymidine incorporation. ATL-2 CM dose-dependently inhibited hepatocyte proliferation. This activity was fractionated by gel filtration at a molecular size of 15,000 to 40,000 and was tentatively called hepatocyte growth inhibitory factor (HGI). Further fractionation with the ion-exchange column indicated that HGI was separable from ADF. Nevertheless, there was a positive correlation between HGI and ADF production, because the HGI activity was also detected in the CM of another ADF producer cell line (HUT102), while no significant HGI activity was detected in the CM of low ADF producer cell lines, ED and MOLT4.

Animals↗

PubMed1991

Elevated serum levels of Dupan-2 in pancreatic cancer patients negative for Lewis blood group phenotype.

CA19-9, a serum marker for pancreatic cancer, gives false-negative results in patients who are negative for the Lewis blood group phenotype. To determine whether other markers may compensate for this drawback, serum levels of CA50, Span-1, sialyl SSEA-1 and Dupan-2 were assayed and compared with those of CA19-9 in 207 normal subjects and in 200 patients with pancreatic carcinoma whose Lewis blood group phenotypes were confirmed. In normal subjects with the Lewis negative phenotype, the serum levels of CA50 and Span-1, as well as CA19-9, were significantly low, whereas those of sialyl SSEA-1 were independent of the Lewis blood group phenotype. Serum levels of Dupan-2 were significantly higher in normal subjects with the Le (a-b-) phenotype as compared with those with Le(a-b+). The sensitivity for pancreatic carcinoma was 81% for CA19-9, 84% for CA50, 82% for Span-1, 51% for sialyl SSEA-1 and 63% for Dupan-2. Among the 39 CA19-9 negative patients, 13 were determined as being Lewis negative by the serum dot-ELISA technique. Although the positive rates were essentially comparable when each marker was combined with CA19-9, a highly elevated serum level of Dupan-2, which strongly suggested the presence of malignancy, was most frequently encountered in 39 patients who were not diagnosed by CA19-9 assay, especially those with Lewis negative blood groups. With regard to the three other markers, we found few patients with a highly elevated serum level in either the Lewis-negative or -positive groups. We conclude that Dupan-2 tended to be elevated in patients with pancreatic cancer who were negative for the Lewis blood group phenotype.

Adult↗

PubMed1991

Preparation of pancreatic cancer-associated mucin expressing CA19-9, CA50, Span-1, sialyl SSEA-1, and Dupan-2.

A new antigen associated with pancreatic cancer was prepared by immunoaffinity chromatography using Fab'-Sepharose beads. This antigen was a glycoprotein of large molecular weight (Mr greater than 8,000,000) in its native state, estimated by size exclusion chromatography on Sephacryl S400. After sodium dodecyl sulfate-polyacrylamide gel electrophoresis and blotting analysis, several cancer-associated glycoconjugates, including CA19-9, CA50, Span-1, Dupan-2, and sialyl SSEA-1, were detected on the antigenic moiety of Mr 90,000. By an enzyme immunoassay for the antigen, elevated levels were found in pooled sera obtained from patients with various malignant and non-malignant diseases and normal subjects. However, the enhanced expression of CA19-9, Lewisa, or Lewisb epitope on the antigen molecule was restricted to the pooled sera from patients with pancreatic cancer. Furthermore, antigens from pancreatic or gastric cancer expressed ligands with intense and specific reactivity for Bauhinia purpurea (BPA), peanut (PNA), and Vicia villosa (VVA) lectins. The present assay system of the antigen, using both monoclonal antibodies (CA19-9, Lewisa, and Lewisb) and lectins (BPA, VVA and PNA), will provide a useful approach to the diagnosis of pancreatic cancer.

Antigens, Neoplasm↗

PubMed1991

Comparative study of CA-50 (time-resolved fluoroimmunoassay), Span-1, and CA19-9 in the diagnosis of pancreatic cancer.

The clinical diagnostic utility of CA-50 (time-resolved fluoroimmunoassay) and Span-1 was compared with that of CA19-9 by measuring their levels in sera from patients with pancreatic cancer and other diseases. In pancreatic cancer CA-50, Span-1, and CA19-9 showed similar positive rates (84%, 82%, and 81%, respectively). With regard to the ability to distinguish pancreatic cancer from chronic pancreatitis, however, the specificity of CA-50 and Span-1 was higher than that of CA19-9 (85%, 85%, and 79%, respectively). Despite the similar positive rates of CA-50 and Span-1 in pancreatic cancer, the correlation between these two markers was low. Thus, used in combination, they compensated for each other in the diagnosis of pancreatic cancer. In chronic liver diseases, serum levels of both CA-50 and Span-1 were correlated with that of biliary tract enzymes, alkaline phosphatase, and r-glutamyl transpeptidase. And these two markers were more affected by the biliary system than CA19-9, resulting in the significantly higher positive rates. In these diseases immunohistochemical study showed that all three markers were localized in the epithelial cells of the bile duct, with CA-50 and Span-1 showing a similar tissue distribution.

Analysis of Variance↗

PubMed1991

Preparation and partial characterization of a pancreatic cancer-associated glycoprotein expressing CA50.

In our previous study, two antigens associated with pancreatic cancer were prepared from ascites of the patients by using a newly developed affinity chromatography technique; one expressed CA19-9, CA50, Span-1, sialyl SSEA-1, and Dupan-2. This report describes the other part of the antigens, which was scarcely found in normal sera. This antigen was a glycoprotein with a high molecular weight of 1,000,000 in its native state, estimated by size exclusion chromatography on Sephacryl S400. After sodium dodecyl sulfate polyacrylamide gel electrophoresis and blotting analysis, the antigenic activity was observed on the 200-Kd band, and monoclonal antibody of CA50 showed intense reactivity to it. By means of an enzyme immunoassay, CA19-9 determinant was also recognized on it to a lesser extent, although sialyl SSEA-1 and Dupan-2 determinants were scarcely observed. From these observations, the present antigen was considered to be a serum glycoprotein carrying type-1 polylactosamine determinant and differs from previously reported glycoprotein carrying CA19-9 in its insolubility in perchloric acid solution because of its lesser degree of glycosilation. Serum levels of the antigen measured by enzyme-linked immunosorbent assay demonstrated that highly positive rates were observed in 78% of pancreatic cancers, 82% of biliary tract cancers, and 96% of hepatocellular carcinomas.

Antigens, Neoplasm↗

PubMed1991

Purification and partial characterization of rat epididymal retinoic acid-binding protein, and its immunohistochemical localization.

Two types of retinoic acid-binding proteins, designated as epididymal retinoic acid-binding protein (ERABP) types A and B, have been purified to homogeneity from rat genital organs. ERABP types A and B had molecular weights of 19,000 and 18,500 and isoelectric points of 5.72 and 5.90, respectively. The absorption spectrum of ERABP complex with retinoic acid had two peaks at 277 and 354 nm, and showed similar uncorrected fluorescence spectra to that of cellular retinoic acid-binding protein (CRABP). These ERABPs were identified as major androgen-dependent epididymal proteins by amino acid sequences analyses. Furthermore, immunohistochemical examinations revealed that the ERABPs exist in the epithelium of the proximal portion of the epididymis and in the lumen of epididymal canal and seminal tract leading from the epididymis. The existence of ERABP strongly suggests that retinoic acid might be involved in the maturation of spermatozoa.

Amino Acid Sequence↗

PubMed1991

Enhancement of rat liver mitochondrial function by portal branch ligation secures subsequent extended hepatectomy.

We have examined the effects of portal branch ligation on liver mitochondrial function and on subsequent extended hepatectomy in rat. In the occluded lobes, mitochondrial function was depressed immediately after the ligation. In the unoccluded lobes, mitochondrial function was enhanced and reached the maximum two days after the ligation. This enhancement was associated with increases in the enzymic activities and subunit amounts of the energy-transducing complexes, and with increase in mitochondrial DNA content. The ligation improved both survival rate and mitochondrial redox state monitored by the ratio of acetoacetate to beta-hydroxybutyrate after hepatectomy. These results suggest that the enhancement of mitochondrial function by portal branch ligation fills the energy demand for liver regeneration.

Animals↗

PubMed1991

Development of an enzyme-linked immunosorbent assay of apolipoprotein E-AII complex in plasma.

Measurement of apolipoprotein (apo) E-AII complex in human plasma is important in determining the role of apoE in lipoprotein metabolism. In this paper, we demonstrate a new and simple method to determine apoE-All complex by using an enzyme-linked immunosorbent assay. Anti-apoE IgG (goat) was used as a capture antibody, and captured apoE-All complexes were detected by an anti-apoAll (rabbit) horseradish peroxidase-conjugated anti-rabbit IgG (goat) system. With this method, apoE-All complex was specifically determined without the interference of apoAll and was not affected by apoE monomer less than 250 mg/L. The content of the complex in reference serum, a normolipidemic serum pooled from five subjects with phenotype E3/E3, was arbitrarily defined as 100%. The coefficients of variation were 3.5%-6.3% within assay and 8.8%-11.6% between assays.

Apolipoproteins E↗