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Y Sakagishi

Publications and source records attributed to Y Sakagishi.

At least 19 recordsLinked to original sources
PubMed1995

Characterization of a specific monoclonal antibody 9F5-3a and the development of assay system for oxidized HDL.

We obtained a monoclonal antibody 9F5-3a against oxidative low-density lipoprotein (LDL) modified with CuSO4 and established a sandwich ELISA for detection of oxidized high-density lipoprotein (oxHDL). The 9F5-3a was reacted strongly with oxHDL and to a lesser degree with oxLDL and LDL. In contrast, little or no reactivity was found with HDL. When the generation of oxHDL was limited by the addition of alpha-tocopherol and catalase, reactivity to 9F5-3a was reduced. Incubation of oxHDL with excess lyso-phosphatidylcholine (lyso-PC) also reduced immunoreactivity, but not by only lyso-PC. These results suggested that the epitope is possibly associated with oxHDL-linked lyso-PC induced mainly by the hydroxyl radical.

Animals↗
PubMed1995

Comparative biochemical study of coated vesicles purified from cock brains.

Cock brain coated vesicles (CBCVs) were purified and compared with porcine brain coated vesicles (PBCVs) from several biochemical aspects. Clathrin heavy and light chains of CBCVs were immunologically similar to those of PBCVs. Coat proteins (CPs) of CBCVs behaved in almost the same manner as those of PBCVs for limited proteolysis. Dissociation of CPs from CBCVs by treatment with 10 mM Tris-Cl, pH 8.5, or 2M urea resembled that of CPs from PBCVs. pH dependency of dissociation of CPs from CBCVs was slightly different from those of PBCVs.

Animals↗
PubMed1995

A variant alkaline phosphatase found in a case of gastric carcinoma with super bone scan.

A rare case of gastric carcinoma associated with increased serum variant alkaline phosphatase activities is presented. A 54 year old man had extremely high serum alkaline phosphatase activity (18,607 U/l) with normal calcium and phosphate concentrations. His bone scintigram showed abnormal findings, 'super bone scan'. He was diagnosed as having Borrmann type 4 gastric carcinoma with diffuse bone metastases by examinations of the upper gastrointestinal tract and iliac bone biopsy. The alkaline phosphatase isozyme of this patient was of the bone type as measured by cellulose acetate membrane electrophoresis and the placenta/bone type by agarose gel electrophoresis, respectively. Immunoelectrophoresis and the immunoprecipitation method using monoclonal antibodies against various alkaline phosphatase isozymes, however, showed that his serum alkaline phosphatase had the liver type antigenicity. Furthermore, it had a larger molecular size and different sugar chains compared with the common liver type alkaline phosphatase. These findings suggest that a unique variant alkaline phosphatase was produced by gastric cancer cells, which is possibly an explanation for the high serum alkaline phosphatase activities in this patient.

Alkaline Phosphatase↗
PubMed1995

[Alanine aminotransferase (ALT)].

Alanine aminotransferase (Alt, L-alanine:2-oxoglutalate aminotransferase) is a pyridoxal enzyme which catalyses the reversible interconversion of L-alanine and 2-oxoglutalate to pyruvate and L-glutamate. The enzyme is widely distributed in various tissues from animals and even in some kind of plants. Isoenzymes of human ALT localize in the cytosol (c-ALT) and mitochondria (m-ALT) of tissues such as liver, kidney, skeletal and cardiac muscles. Amino acid sequence of c-ALT from rat and human liver has been wholly determined by Ishiguro et al. It is suggested that c-ALT is associated to the utilization of pyruvate in glycolysis and m-ALT is involved in the conversion of alanine to pyruvate for gluconeogenesis.

Alanine↗
PubMed1994

Carbohydrate-mediated recognition of a circulating placental alkaline phosphatase-immunoglobulin M complex.

We detected an abnormal alkaline phosphatase (AP) electrophoretically in the serum of a patient with rheumatoid arthritis, who had a macromolecular AP linked with immunoglobulin M (IgM) bearing a kappa light chain. The IgM isolated from the AP-IgM complex in the patient's serum reacted apparently with all of the AP isozymes tested, i.e. those originating in the liver, bone, intestine and placenta, but the alpha-mannosidase-treated IgM from the patient's serum bound to placental AP (PAP) alone. This suggests that untreated IgM recognizes multivalent epitopes of the AP and that the complex of AP with alpha-mannosidase-treated IgM is a specific antibody-antigen complex. In order to investigate further the multivalent binding capacity for the PAP-untreated IgM complex, we prepared a monoclonal antibody (MoAb) against PAP and identified it as an IgM with a kappa light chain. The binding affinities and their circulating half-lives of the synthetic complexes of PAP and respective MoAbs were examined with and without treatment with several glycosidases. The untreated MoAb bearing IgM had binding affinity for all of the AP isozymes tested, while alpha-mannosidase-treated IgM attached only to PAP, the same as the IgM isolated from the PAP-IgM complex in the patient's serum. The circulating clearance of the PAP-IgM complex in rabbits was faster than either component alone. In addition, the PAP-IgM complex treated with alpha-mannosidase was found to have the shortest half-life of all the complexes of PAP and Igs treated with the several glycosidases tested. These results suggest that the formation of the PAP-IgM complex as an enzyme-linked antibody and the clearance of the complex in vivo are dependent on the sugar moieties of the Igs.

Adult↗
PubMed1994

Deoxyuridine triphosphatase in human hepatoma.

1. Deoxyuridine triphosphatase (dUTPase) in human hepatoma was investigated. The apparent activity in a gram weight tissue was about 6 times that of the activity in rat livers after partial hepatectomy. 2. Most catalytic properties of the hepatoma dUTPase were similar to the enzymes from other sources. 3. The hepatoma dUTPase reacted with the antibodies against the rat spleen dUTPase, although the molecular size of the enzyme (46-48 kDa) was different to the rat enzyme. 4. The immunochemical studies indicated that the enzyme was composed of two identical subunits, whose molecular mass was about 22 kDa.

Animals↗
PubMed1994

Phosphorylcholine as a unique substrate for human intestinal alkaline phosphatase.

1. The enzymatic nature of human liver, bone, placental and intestinal alkaline phosphatases (ALPs) were investigated with phosphorylcholine (PC), phosphorylethanolamine, pyridoxal-5'-phosphate and p-nitrophenylphosphate at a weakly alkaline pH. 2. The apparent Km value of the intestinal ALP with PC was the highest of all ALPs tested. Intestinal ALP hydrolyzes PC the most and has higher affinity for choline as a transphosphorylating acceptor than the other ALPs. In addition, the intestinal ALP activity with PC was most susceptible to Na2HPO4, in the tested ALPs. 3. The present results suggest that PC is a unique substrate for human intestinal ALP, which may be related to the metabolism of PC or choline as part of phosphatidylcholine.

Alkaline Phosphatase↗
PubMed1994

Diazotization reaction of nitric oxide trapped by hemoglobin.

The present study addresses to determine whether hemoglobin within red blood cells can be utilized as a spin-trap agent for nitric oxide. We demonstrate the diazotization method coupled with a gel filtration chromatography, which is simply due to the separation of nitrosylhemoglobin from nitrite, nitrate or other low molecular nitroso-compounds in biological systems and to the liberation of nitric oxide from nitrosyl heme-complexes in the acidic condition. The amount of nitric oxide can be estimated by the difference of absorbances at 542 nm between diazo-compounds formed by Griess reagent and hemichrome by phosphoric acid. Our results indicate that hemoglobin in red cells as a spin-trap agent would be useful for monitoring nitric oxide in the circulation under the several disease states.

Chromatography, Gel↗
PubMed1994

Sugar chain heterogeneity of human urinary chorionic gonadotropin determined by serial lectin affinity chromatography: difference between benign and malignant disease.

Human chorionic gonadotropin (hCG) is a glycoprotein of which sugar chains are considered to show structural changes with malignancy. To study the sugar chain heterogeneity of urinary hCG in patients with gynecological disease, we employed serial lectin affinity chromatography (LAC) using concanavalin A (Con A) and phytohemagglutinin-E (PHA-E) which can separate N-glycoside-linked sugar chains, and Jacalin lectin which is specific for O-glycoside-linked sugar chains. The proportion of hCG which did not bind to Con A was clearly higher in patients with cervical cancer than in healthy pregnant women. The complex-type sugar chains bearing bisecting (beta 1-4) N-acetylglucosamine which bound to PHA-E increased in the early stage of cervical cancer, and tri- and tetra-antennary complex type sugar chains also increased in the advanced stages. In addition, the Jacalin-bound hCG increased significantly along with the stage of the cancer, especially in advanced cervical cancer with distant metastases. Taken together, these results show that alteration in sugar chain structures of hCG reflect the advanced stage of cervical cancer.

Carbohydrates↗
PubMed1994

Differences between the sugar moieties of liver- and bone-type alkaline phosphatases: a re-evaluation.

We re-evaluated the differences between the sugar moieties of liver and bone alkaline phosphatases (ALPs). Sialic acid was added to ALP sugar moieties by alpha 2,3- or 2,6-sialyltransferase treatment of the asialo-form ALP (neuraminidase-treated ALP). Asialo-bone ALP was converted to a liver-like ALP by the 2,6-sialyltransferase treatment. The resulting liver-like ALP was less susceptible to neuraminidase than non-treated bone ALP, but was still labile to heat exposure at 56 degrees C like non-treated bone ALP. However, after the O-linked sugar moiety had been released by additional treatment with O-glycanase the liver-like ALP became more heat stable at 56 degrees C, like non-treated liver ALP. Non-treated liver ALP reacted specifically with anti-liver ALP monoclonal antibody, and non-treated bone ALP reacted with both anti-liver and anti-bone ALP antibodies. The asialo-bone ALP still reacted with anti-bone ALP antibody, whereas the asialo-form liver ALP showed little, if any, reaction with anti-liver and anti-bone ALP antibodies. Neuraminidase and O-glycanase-treated bone ALP reacted less with anti-bone ALP antibody. After O-glycanase treatment, bone ALP molecules deprived of an O-linked sugar moiety had a molecular size and heat stability similar to liver ALP. The difference between liver and bone ALP molecules may be due not only to their manner of sialic acid linkage but also to the attachment of the O-linked sugar moiety.

Alkaline Phosphatase↗
PubMed1992

Differences in the enzymatic nature and the sugar-chain structure of gamma-glutamyl transferase between normal and carcinomatous human kidney and prostate.

The enzymatic and immunological nature, and the sugar chain structure, of gamma-glutamyl transferase (GGT) purified from tissues of benign prostatic hypertrophy (BPH), prostatic carcinoma (PCa) and renal cell carcinoma (RCa), were compared with those of the normal prostate (NP) and kidney (NK). The specific activities of GGTs in NP, NK, BPH, PCa and RCa were 78.9, 22.5, 105, 92.5 and 52.5 mU/mg protein, respectively. The molecular masses of GGTs from BPH, PCa and RCa were 72 kDa, 78 and 108 kDa, and 79 and 105 kDa, respectively. The Michaelis constants (Km), optimum pHs and the inhibition of GGT activities by several chemical compounds, revealed that the GGT from BPH, PCa and RCa was similar to that of normal GGT. Immunologically, the IgG fraction against anti-human seminal plasma GGT fused to the all of the GGTs tested. The sugar chain heterogeneities of the various GGTs, detected by the serial-lectin affinity technique, differed from one another. The sugar chain of GGT from BPH resembled the sugar chain from NP. On the contrary, the sugar chains of GGTs from PCa and RCa were markedly different from those from normal tissues. In the GGT from PCa, multi-antennary complex type sugar chains were more increased than the enzyme of NP. In general, as previously reported, the sugar chains of GGTs from carcinomatous tissues of prostate and kidney had an increased content of bisecting GlcNAc (beta 1-->4) containing complex type sugar chains. Moreover, the reductions of the biantennary complex type sugar chain with fucose linkage and the hybrid type sugar chain were obvious in the GGT from carcinomatous tissues of the prostate and kidney.

Adult↗
PubMed1992

Intestinal type alkaline phosphatase hyperphosphatasemia associated with liver cirrhosis.

Hyperphosphatasemia due to increased intestinal type serum alkaline phosphatase was noted in a 48-year-old male who had asymptomatic liver cirrhosis. The alkaline phosphatase activity in the serum was 828 U/l (our reference range in adults: 57-194 U/l), 94% of which was of the intestinal type as measured by an immunoprecipitation method. The intestinal component of alkaline phosphatase was separated into two major and some minor components using electrophoresis and isoelectrofocusing. One of the major components had similar mobility to that of a standard intestinal enzyme purified from adult intestine. The components were heat-labile and neuraminidase-resistant. Serial lectin affinity chromatography, however, indicated that sugar chain compositions of the alkaline phosphatase were different from those of the standard tissue intestinal enzyme. These results and further enzymological studies suggest that the patient's serum alkaline phosphatase basically consisted of several intestine-like isoforms.

Alkaline Phosphatase↗
PubMed1992

Purification and characterization of cytoplasmic 5'(3')-nucleotidase from rabbit spleen: characteristic differences of the enzyme from the rat spleen nucleotidase.

1. A 5'(3')-nucleotidase, which preferably hydrolyzed 3'-dTMP and 3'-dUMP, was highly purified from rabbit spleen cytosol. 2. The enzyme also hydrolyzes 3'-UMP, 5'-dUMP and guanine nucleotides, but does not hydrolyze any adenine nucleotides or cytosine nucleotides. 3. The activity is dependent upon Mg2+, Co2+ or Mn2+; the addition of deoxyinosine stimulates the activity, and the pH optimum for the hydrolysis of 3'-dTMP is 7.0. 4. Although the catalytic properties of the enzyme are similar to the 5'(3')-nucleotidase from rat spleen, these nucleotidases differ in their molecular disposition. 5. The charge state of the rabbit enzyme is slightly basic, and the subunit M(r) is about 27 kDa, while the value of the rat enzyme is 26 kDa. 6. Immunochemical experiments with the use of antibodies against the purified nucleotidase indicate that enzymes from rabbit spleen and from rat spleen are composed of different polypeptides.

Animals↗
PubMed1992

[Estimation of peripheral metabolism under cardiopulmonary bypass by measuring ketone bodies, lactate, and pyruvate].

Both ketone body ratio (KBR) and pyruvate/lactate ratio (P/L) are metabolic indicators related to NAD-linked dehydrogenase system according to the REDOX THEORY. In order to estimate the peripheral metabolic state under the cardiopulmonary bypass (CPB) which is considered to be a moderate and controllable shock state, we measured the pyruvate (P), lactate (L) and ketone bodies (acetoacetate [ACA], 3-hydroxybutyrate[HOB]) pre, during, and post-CPB in 20 open heart surgical patients (Group 1). We also measured ketone bodies in another group (Group 2: 22 patients) of elective coronary artery bypass surgery (CABG). Five patients in Group 2 who developed postoperative organ failure (Group 2-A) were compared with the other 17 cases with uneventful postoperative course (Group 2-B). In Group 1, P and L concentrations progressively increased until the end of CPB, thereafter decreased (P less than 0.001). HOB and TKB significantly increased immediately after the start of CPB, afterward which declined during CPB and returned to the control level after CPB. Both KBR and P/L regression analysis reveal the significant correlation between P/L and KBR (r = 0.51, P less than 0.001). KBR at 1 hour after CPB returned to the control level, however, P/L at 1 hour after CPB still stayed in the significant lower level than the control (P less than 0.001). KBR at the aortic declamping in group 2-A was significantly lower than that in group 2-B. In conclusion, P/L and KBR changed in relation to hemodynamic change during CPB.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1991

Liver-like alkaline phosphatase in the tissue-unspecific type enzyme found in rabbit organs.

Rabbit liver and kidney tissues are known to produce an intestinal-like alkaline phosphatase (IAP-like enzyme) as a dominant isozyme, with a minor isozyme of tissue-unspecific type (UAP), unlike humans and other mammalians. We investigated immunohistochemically and biochemically these unique isozymes in the rabbit liver and bone, and compared them with the human isozyme. In rabbit liver, UAP was found to be localized only in the apical part of the membrane of cells lining the bile duct, whereas IAP-like enzyme was found in the sinusoidal membrane of hepatocytes. Rabbit liver UAP was separated from IAP-like enzyme by DEAE-cellulose column chromatography. Rabbit bone tissue contained only one UAP isozyme. The two UAPs were biochemically and physicochemically compared with human liver AP. Both UAPs reacted with an anti-human liver AP monoclonal antibody, not with an anti-human bone AP monoclonal antibody, indicating that both enzymes have the same antigenicity as human liver AP. Rabbit liver and bone UAPs had similar N-linked sugar-chain heterogeneities to the respective human enzymes. In addition, rabbit bone AP also had an O-linked sugar chain, as did human bone AP, unlike rabbit and human liver APs.

Alkaline Phosphatase↗
PubMed1991

Purification and partial characterization of intestinal-like alkaline phosphatase in rabbit kidney.

Two types of alkaline phosphatase (AP) isozymes in rabbit kidney, a major intestinal-like type and a minor tissue-unspecific type, have been identified. The former enzyme was purified from rabbit kidney by immunoaffinity chromatography using monoclonal anti-human intestinal AP antibody. The purified enzyme yielded a single protein band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the apparent molecular size of its monomer subunit was found to be 72,000. Three amino acid residues within the first 16 N-terminal amino acid residues were different in purified AP and human intestinal AP. Although the rabbit enzyme possessed some peptide bands identical to those of human adult intestinal AP after Staphylococcus aureus V8 protease digestion, the enzyme did not react with monoclonal antibody against human adult intestinal AP alone, whereas it did react with monoclonal antibody against both human adult and fetal intestinal APs. The affinity of the enzyme for concanavalin A was identical to that of the fetal intestinal AP, but different from that of the adult enzyme. These results indicate that the antigenicity and certain properties of purified rabbit AP are more like those of human fetal intestinal AP or Kasahara isozyme, so-called intestinal-like AP, than like human adult intestinal AP.

Alkaline Phosphatase↗
PubMed1991

[Relationship among glycated compounds, superoxide dismutase activities, and other related analytes in diabetic patients classified by ages].

Among several glycated compounds (GC) which are based on Maillard reaction, glycated hemoglobin (HbA1c) and fructosamine (FRA) have been utilized widely as a markers of diabetes. Recently, glycated albumin (GA) has been pointed out as a new indicator. For the determination of GA, spectrophotometry combined with an affinity column method has been mainly used, however the procedure is complicated. Recently a two-column HPLC method (ion-exchange column and affinity column) has been developed by Shima. We have evaluated a GA analyzer GAA-2000 based on Shima's method. After a series of fundamental and performance evaluation studies, the GAA-2000 was found to be appropriate for our study. Reference values obtained from this equipment were 10.56-16.87%. Correlation coefficients based on GA using diabetic and diabetic nephropathy patient specimens (n = 87) were: FRA (r = 0.944) greater than HbA1c (r = 0.842) greater than Glucose (r = 0.510) Superoxide dismutase (SOD) and lipid peroxidase (LPO) with are produced in relation to active oxygen did not show a good correlation. Although we tried classify the patients according to juvenile (20-39), middle (40-64) and senile (greater than 65) the method of Asada et al., we could not find any distinct tendencies.

Aged↗