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T Komoda

Publications and source records attributed to T Komoda.

At least 55 records · Page 3Linked to original sources
PubMed1997

Human tissue non-specific alkaline phosphatases: sugar-moiety-induced enzymic and antigenic modulations and genetic aspects.

To investigate the possible role(s) of glycans in human tissue non-specific alkaline phosphatase (TNAP) activity, the iso-enzymes were purified and treated with various exo- and endo-glycosidases. Catalytic activity, oligomerization, conformation and immunoreactivity of the modified TNAPs were evaluated. All TNAPs proved to be N-glycosylated, and only the liver isoform (LAP) is not O-glycosylated. Usually, the kidney (KAP) and bone (BAP) isoenzymes are similar and cannot be clearly discriminated. Differences between the immunoreactivity of KAP/BAP and LAP with a BAP antibody were mainly attributed to the N-glycosylated moieties of the TNAPs. In addition, elimination of O-glycosylations moderately affects the TNAP reactivity. Interestingly, N-glycosylation is absolutely essential for TNAP activity, but not for that of the placental or intestinal enzymes. According to the deduced amino acid sequence of TNAP cDNA, Asn-213 is a possible N-glycosylation site, and our present findings suggest that this sugar chain plays a key role in enzyme regulation. With regard to the oligomeric state of alkaline phosphatase (AP) isoforms, the dimer/tetramer equilibrium is dependent on the deglycosylation of glycosyl-phosphatidylinositol(GPI)-free APs, but not GPI-linked APs. This equilibrium does not affect the AP conformation as observed with CD. With regard to TNAPs, no data were available on the gene expression or nature of the 5'-non-translated leader exon of human KAP, as opposed to BAP and LAP genes. cDNA sequencing revealed that cortex/medulla KAP is genetically related to BAP, and medulla KAP to LAP.

Alkaline Phosphatase↗
PubMed1997

Intestinal alkaline phosphatase isoforms in rabbit tissues differ in glycosylation patterns.

OBJECTIVES: In patients with advanced liver cirrhosis or chronic nephritis, an intestinal alkaline phosphatase (IAP)-like enzyme is ectopically expressed in the liver or kidney. In this study, we used rabbit organs as a human pathological model, because the rabbit liver or kidney expresses an IAP-like enzyme as the predominant isozyme, unlike humans. METHODS: IAP and the IAP-like enzyme were purified from rabbit intestine and kidney, respectively, by immunoaffinity chromatography using a monoclonal antihuman IAP antibody. Some properties of the IAP and IAP-like enzyme expressed in rabbit organs are compared. RESULTS: Some of their catalytic and physicochemical properties differed. In particular, the net charge, molecular mass, and hydrophobicity of IAP from rabbit intestine was slightly different from the IAP-like enzyme from rabbit kidney. There was a difference in the sugar chain structure between the two enzymes according to the results of lectin affinity chromatography, and part of the peptide maps differed slightly. However, there was no difference in the peptide maps after treatment with endo-N-acetylglucosaminidase F. CONCLUSIONS: The primary structures of IAP and the IAP-like enzyme are basically similar, except for the glycosylation process of the respective AP isozymes.

Alkaline Phosphatase↗
PubMed1997

Imaging of the orifice of the left ventricular outflow tract: technique and initial results.

The mechanism of left ventricular outflow tract (LVOT) obstruction in the patient after mitral valve replacement or repair was examined with the aid of 2D echocardiography. For the interpretation of the spatial relationship between the aortic root and mitral annulus, however, the 2D display is sometimes inadequate since it may not simultaneously capture these structures in each center. We developed a method to clarify this relationship in 3D based on magnetic resonance images. We defined the office of the left ventricular outflow tract (LVOT orifice), consisting of, in turn, a muscular region, i.e., edge of the interventricular septum, and an annular region, i.e., the annulus of the anterior mitral leaflet. In this paper we present image data obtained from one of eight normal subjects examined and compare this with data of one of two patients who preoperatively suffered degenerative mitral regurgitation and subsequently underwent chordal-preserving mitral valve replacement, in which anterior chordae were reattached to the anterior annulus. In the normal subject, the mitral annulus exhibited a flexible change in shape during the systole, maintaining sufficient LVOT orifice size. In the patient, the prosthetic valve exhibited translational motion during systole, resulting in dynamic LVOT obstruction. This phenomenon was also observed in one other case. Furthermore, the lateral view of the LVOT orifice revealed a projection of the prosthetic valve into the LVOT, causing mechanical LVOT obstruction. The finding that translational motion of the prosthetic valve with an inflexible mitral annulus results in dynamic LVOT obstruction may support the hypothesis that annular rigidity causes dynamic LVOT obstruction after mitral valve repair with a rigid prosthetic ring.

Adult↗
PubMed1996

[Detection of chlamydial antibodies by immunoblotting technique--in comparison of three commercially available test kits and micro-IF].

We reexamined the assay conditions of immunoblotting (W-B) technique to detect antibodies (IgG and IgA) to C. trachomatis serovar L2. Partially purified Chlamydia (35% urografin) was used as the antigen. The marker bands of W-B for positive and/or negative are major outer membrane protein (MOMP) band and either one or more bands staining in 40-62 KDa area. We then compared the sensitivity and specificity of three commercially available test kits and micro-IF by using W-B as a standard. The kits compared were sero-IPALISA-IgG-IgA, IP-Azyme-IgG-IgA and HITAZYME-IgG-IgA, and micro-IF. Serum samples were collected from the outpatient departments of gynecology, urology, and internal medicine and pediatrics. The results of agreement between W-B and test kits in IgG detection were as follows: in sero-IPALISA, total agreement was 85.7%, positive agreement 83.3%, negative agreement 90.9% and in IPAzyme: 85.7%, 83.3%, 90.9%, respectively and in HITAZYME: 82.9%, 75%, 100%, respectively. These results are almost the same as micro-IF: 85.7%, 79.2%, 100%, respectively. These kits may have relatively high sensitivity and specificity in IgG antibody detection. In IgA detection, the total agreement between W-B and sero-IPALISA was 82.9%, positive agreement 100%, negative agreement 66.7%, in IPAzyme: 77.1%, 58.8%, 94.4%, respectively and in HITAZYME: 65.7%, 64.7%, 66.7%, respectively and in micro-IF: 82.9%, 70.6%, 94.9%, respectively. Although these agreements are not so high in IgA detection, the W-B technique gives fairly consistent results as well as those kits. This indicates that W-B technique with MOMP and other protein bands (40-62 KDa) as marker for positive reaction is a useful method for detection of chlamydial antibodies.

Antibodies, Bacterial↗
PubMed1996

A variant alkaline phosphatase detected in a patient with lung cancer.

A variant alkaline phosphatase (ALP), with heat-sensitivity characteristics similar to that of the bone type, was found in the serum of a patient suffering from lung cancer. In disc polyacrylamide gel electrophoretic studies most of this enzyme had migrated to the region corresponding to liver ALP, with the remainder affecting bone ALP. Like kidney ALP, this ALP was markedly inhibited by 0.5 mmol/l L-cysteine. The K(m) of this ALP for p-nitrophenylphosphate was 0.39 mmol/l, similar to that of kidney ALP. The sugar moiety of this enzyme bore greater resemblance to that of kidney ALP than liver or bone ALP. However, immunoprecipitation of this particular ALP was strong with a monoclonal antibody against liver ALP and moderate with an antibody against bone ALP.

Aged↗
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

A precursor form of human kidney gamma-glutamyl transferase in normal and cancerous tissues, and its possible post-translational modification.

We have found three molecular forms of human gamma-glutamyl transferase (GGT) in normal and renal cell carcinomatous tissues, and also have reported the marked differences in the sugar chains of GGTs between normal and cancerous tissues by serial lectin affinity chromatographies. In this study, the peptide maps of three purified GGTs (79 kDa, 50 kDa and 25 kDa) obtained by lysylendopeptidase digestion, the subcellular localization of GGTs, and the sugar chains of GGTs were compared between normal and cancerous tissues. According to the results, the total peptide bands of the digested 79 kDa component represented the sum of those of the digested 50 and 25 kDa components on 12.5% SDS-PAGE. In addition, C-terminal and N-terminal amino-acid sequences of the 79 kDa protein were the same as the sequences of light and heavy subunits, respectively, suggesting that the 79 kDa component is of the precursor form of the 50 kDa mature heavy and 25 kDa light subunits, respectively. On the other hand, the GGT activity in renal cell carcinomatous tissues was significantly increased in the microsomal fraction and decreased in the soluble fraction compared with that of normal tissues. Meanwhile, the sugar moiety of GGTs in the respective subcellular fractions was obviously different between normal and cancerous tissues. In particular, a reduced multiantennary complex type sugar chain and an elevated high-mannose or hybrid-type sugar chain in the microsomal fraction were observed in the GGT in cancerous tissues.

Amino Acid Sequence↗
PubMed1995

Study of cardiac structures in relation to the pericardial cavity for total artificial heart implantation.

To enhance the design of the completely implantable total artificial heart (TAH), we developed a coordinate system that expresses the geometry of the heart, including that at the interface with the TAH and that of the pericardial cavity. This coordinate system (ventricular coordinate system) was developed using 3D images of the heart which were obtained from magnetic resonance images. The origin is located on the centroid of the mitral annulus at 25 ms delay from the ECG R wave (M0), and the y axis is the axis from M0 to the apex at 25 ms delay (Apex0). The x-y plane includes M0, Apex0, and the centroid of the tricuspid annulus at 25 ms delay. Since the y axis expresses the apical orientation of the heart, data using this coordinate system are not affected by individual variation in heart orientation. Heart geometry data were obtained in 5 normal subjects. Compared with that using the coordinate system based on the thoracic wall, the expression of the data of interface geometry for the heart and the TAH is simplified when the ventricular coordinate system is used. A method of expressing the geometry of the pericardial cavity using the ventricular coordinate system is also proposed. These methods may be applicable in obtaining data in patients suffering from cardiomyopathy.

Adult↗
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

USF-19A, a new lipoxygenase inhibitor from Streptomyces sp.

USF-19A, a soybean lipoxygenase (SBL) and human 5-lipoxygenase (5-LO) inhibitor, was isolated from Streptomyces sp. USF-19 strain. Its chemical structure was determined by spectroscopic evidence to be a new member of the antimycin antibiotic family. The IC50 value of USF-19A against 5-LO was 28.0 microM.

Benzamides↗
PubMed1995

[Multiple forms of certain enzymes result post-translationally by a modification of sugar or protein moieties].

Multiple forms of a certain enzyme may result from at least two mechanisms: first, allozymes are coded by distinct genes which exist in separate locus on the chromosome or processed by shuffling of distinct exons on the same genome, and second is this subject, so-called isozymes biosynthesized from single gene subsequently become distinguishable from each other as a result of post-translational modification by protease cleavage (CK), deamidation (AMY) and sugar (ALP, GGT, AMY) or GPI-anchor (ALP) moieties attaching to the enzyme molecules. Therefore, it is interesting to speculate whether alternative forms of the above-mentioned enzymes are true isozymes synthesized from a mRNA from single and/or different cells. In this section, the current topics of these isozymes are commented or discussed.

Carbohydrate Sequence↗
PubMed1995

Preservation of papillary muscles and chordae during mitral valve replacement: possibilities and limitations.

The various techniques of preserving the chordal apparatus in mitral valve replacement applied in 1,453 cases at a single institution between 1986 and 1994 are described. The techniques include preservation of chordal attachment to the posterior mitral leaflet only and to both leaflets with five different technical variations. The specific advantages and risks of these techniques are demonstrated and discussed. It is concluded, that preservation of the entire subvalvular structures is feasible in almost all cases. Different procedures should be employed according to the variety of mitral disease, in particular with respect to mitral leaflet mobility.

Chordae Tendineae↗
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

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↗