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K Azumi

Publications and source records attributed to K Azumi.

At least 19 recordsLinked to original sources
PubMed1997

Cloning and tyrosine phosphorylation of a novel invertebrate immunocyte protein containing immunoreceptor tyrosine-based activation motifs.

Immunoreceptor tyrosine-based activation motif (ITAM) plays an important role in signal transduction through mammalian T-cell and B-cell antigen receptors and Fc receptors. The ITAM has been found only in vertebrate immunocytes. Ascidians are intriguing invertebrates from the viewpoint of the evolution of immune systems because they are considered to be ancestors of the vertebrates. We have previously shown that the monoclonal antibody A74 inhibits cellular defense reactions of the ascidian. In the present studies, we found that the A74 antigen protein has two ITAMs and several motifs that are proposed to function in signal transduction. The A74 protein is tyrosine-phosphorylated and associated with other proteins in the initial stages of cellular defense reactions. The ITAMs of the A74 protein are tyrosine-phosphorylated by a c-Src kinase in vitro. The A74 protein provides a key to the understanding of the origin of vertebrate immune systems.

Amino Acid Sequence↗
PubMed1997

Ancient origin of the complement lectin pathway revealed by molecular cloning of mannan binding protein-associated serine protease from a urochordate, the Japanese ascidian, Halocynthia roretzi.

Recent identification of a C3-like gene in sea urchins revealed the presence of a complement system in invertebrates. To elucidate further the components and function of the pre-vertebrate complement system, we attempted to isolate an ascidian (urochordata) C3 convertase. After identification of C3 cDNA from Halocynthia roretzi, a Japanese ascidian, reverse transcriptase-PCR amplification of hepatopancreas RNA was performed using primers encoding highly conserved amino acid sequences of the vertebrate Bf and C2 serine protease domain. Two candidate sequences were identified, and the corresponding cDNA clones were isolated from a hepatopancreas library. Surprisingly, neither clone is related to Bf/C2 but rather share the same domain structure of mammalian C1r/C1s/MASP (mannan binding protein-associated serine protease), and are more related evolutionarily to mammalian MASP than to mammalian C1r or C1s. The identification of the tunicate MASP clones, amplified with primers designed to amplify Bf or C2, suggests that the lectin pathway antedated the classical and alternative pathways of complement activation.

Amino Acid Sequence↗
PubMed1996

Unexpected beta2-microglobulin sequence diversity in individual rainbow trout.

For mammals beta2-microglobulin (beta2m), the light chain of major histocompatibility complex (MHC) class I molecules, is invariant (or highly conserved) and is encoded by a single gene unlinked to the MHC. We find that beta2m of a salmonid fish, the rainbow trout (Oncorhynchus mykiss), does not conform to the mammalian paradigm. Ten of 12 randomly selected beta2m cDNA clones from an individual fish have different nucleotide sequences. A complex restriction fragment length polymorphism pattern is observed with rainbow trout, suggesting multiple beta2m genes in the genome, in excess of the two genes expected from the ancestral salmonid tetraploidy. Additional duplication and diversification of the beta2m genes might have occurred subsequently. Variation in the beta2m cDNA sequences is mainly at sites that do not perturb the structure of the mature beta2m protein, showing that the observed diversity of the trout beta2m genes is not primarily a result of pathogen selection.

Amino Acid Sequence↗
PubMed1996

Characterization of novel metallo-proteases released from ascidian hemocytes by treatment with calcium ionophore.

We have previously demonstrated that calcium ionophore induced the release of a novel metallo-protease from hemocytes of a solitary ascidian, Halocynthia roretzi. Here, we isolated the enzymes, PI and PII, from the culture media of H. roretzi hemocytes, which had been treated with calcium ionophore, A23187. The purification procedure included hydrophobic and anion-exchange chromatographies, and gel filtration. The molecular weights of the enzymes were estimated to be 11,000 by gel filtration, but the apparent sedimentation coefficients were 5.0 S, which suggests that the H. roretzi enzymes are of larger proteins with molecular weights of 80,000-90,000. The most susceptible substrate was succinyl-Leu-Leu-Val-Tyr-4-methylcoumaryl-7-amide, and the optimum pH was 8.0, in either case of PI or PII. The activities of PI and PII enzymes were strongly inhibited by metal-chelating agents and propioxatin A, but not by phosphoramidon, a typical metallo-protease inhibitor. Zinc and calcium ions were found to be essential for the maximum expression of protease activity in both enzymes. Thus, the isolated enzymes are characterized as phosphoramidon-insensitive metallo-proteases, which are inhibited by propioxatin A. Extracellular roles of these enzymes were also discussed.

Animals↗
PubMed1995

A novel membrane glycoprotein involved in ascidian hemocyte aggregation and phagocytosis.

Invertebrate hemocytes undergo several cellular defense reactions. To clarify the molecular mechanisms for cellular recognition between hemocytes and also between hemocytes and foreign materials, we established hybridoma clones producing monoclonal antibodies that inhibit hemocyte aggregation (i.e. a cellular reaction between hemocytes) in the solitary ascidian, Halocynthia roretzi. The antibody, A74, also inhibited phagocytosis of foreign materials by H. roretzi hemocytes. Immunocytochemistry of H. roretzi hemocytes using A74 antibody revealed the localization of the A74 antigen on the surface of hemocytes. The A74 antigen, which is referred to as A74 protein, was purified from a hemocyte membrane preparation by three chromatographies on phenyl-Sepharose, A74 antibody-immobilized Sepharose and Mono Q. The A74 protein was a glycoprotein with a molecular mass of 160 kDa; N-glycosidase or neuraminidase treatment resulted in a reduction of its molecular mass. The N-terminal amino acid sequence of A74 protein showed little similarity to other known proteins. Thus, the A74 protein is a novel membrane protein that plays an important role in ascidian cellular defense reactions.

Amino Acid Sequence↗
PubMed1994

Hemocyte aggregation in the solitary ascidian Halocynthia roretzi: plasma factors, magnesium ion, and Met-Lys-bradykinin induce the aggregation.

Hemocytes of the ascidian Halocynthia roretzi undergo aggregation in hemolymph that has been collected from the body through the tunic. To investigate the mechanisms involved, we first established two methods of measuring hemocyte aggregation. In one method, hemocyte aggregation was quantified by its reduction of light scattering intensity as measured with a fluorescence spectrophotometer. In the other method, the increase of transmittance accompanying aggregation was measured with an ELISA reader. We found that ascidian plasma, Mg2+, and Met-Lys-bradykinin can induce the hemocytes of H. roretzi to aggregate. The aggregation induced by any of these three substances was inhibited by EDTA, N-ethylmaleimide, and cytochalasin B. Lipopolysaccharide had little inducing effect. We also demonstrated that, when H. roretzi plasma was treated with trypsin, low molecular weight aggregation-inducing substances were produced. These results suggest that metal ions and peptide-like substances present in the hemolymph play essential roles in the progression of hemocyte aggregation of H. roretzi.

Amino Acid Sequence↗
PubMed1991

Lipopolysaccharide induces release of a metallo-protease from hemocytes of the ascidian, Halocynthia roretzi.

Hemocytes of the solitary ascidian, Halocynthia roretzi, released a succinyl-Leu-Leu-Val-Tyr-4-methylcoumaryl-7-amide hydrolyzing enzyme in response to lipopolysaccharide treatment. The response was dependent on the temperature for incubating hemocytes. The protease release reaction was not triggered by beta 1-3 glucan. The protease released showed strict substrate specificity and its activity was inhibited by EDTA and o-phenanthroline, but not by phosphoramidon, diisopropylfluorophosphate, N-ethylmaleimide, or p-chloromercuribenzoic acid. Thus, the enzyme was characterized as a phosphoramidon-insensitive metallo-protease. Calcium ionophore, phorbol myristate acetate, concanavalin A, and thrombin also induced the release of the same protease from H. roretzi hemocytes.

Amino Acid Sequence↗
PubMed1991

A novel lipopolysaccharide-binding hemagglutinin isolated from hemocytes of the solitary ascidian, Halocynthia roretzi: it can agglutinate bacteria.

A hemagglutinin was isolated from hemocytes of the ascidian, Halocynthia roretzi, by a procedure including extraction and ion-exchange chromatography on CM-cellulose. The molecular weight of the hemagglutinin was estimated to be 120,000 by gel filtration. It was resistant to acid treatment but sensitive to alkali or heat treatment. The hemagglutinating activity was inhibited by heparin, chondroitin sulfate, and lipopolysaccharide (LPS), but not by mono- and disaccharides such as N-acetyl-galactosamine, galactose, and melibiose. The hemagglutinin showed binding ability to heparin and LPS, as demonstrated by heparin-Sepharose chromatography and centrifugation experiments, respectively. It was also found that the hemagglutinin can bind to various bacteria such as Escherichia coli, Bacillus subtilis, Vibrio anguillarum, Pseudomonas perfectomarinus, Achromobacter aquamarinus, and Alteromonas putrefaciens, and can agglutinate all of them.

Animals↗
PubMed1991

[Successful repair of subvalvular annular aneurysm of the left ventricle].

A 62-year-old woman was admitted to our hospital due to congestive heart failure. On the chest X-ray film, cardiomegaly and prominent pulmonary congestion were revealed. She became well with bed rest and medication of digitalis and diuretics. Echocardiogram established the diagnosis of severe mitral regurgitation. In addition, cineangiography of the left ventricule showed cystic left ventricular aneurysm which originated from below the posterior mitral annulus. Operation was performed on July 16, 1990. Under cardiopulmonary bypass, we reflected the apex of the LV upwards, but failed to identify the aneurysm. Then, left atrium was opened through Dubost incision. As regards mitral valve structures, there were no abnormal findings except moderate enlargement of it's orifice. After resection of the mitral valve, orifice of the aneurysm (0.5 X 2.0 cm) was recognized, which was closed with the buttressed sutures. Afterwards, MVR was performed with 25 mm Medtronic Hall valve. The patient had a smooth postoperative course without complication.

Female↗
PubMed1990

Inhibitory effect of halocyamine, an antimicrobial substance from ascidian hemocytes, on the growth of fish viruses and marine bacteria.

Halocyamine A, an antimicrobial substance isolated from hemocytes of the solitary ascidian Halocynthia roretzi, inhibited in vitro the growth of fish RNA viruses (infectious hematopoietic necrosis virus and infectious pancreatic necrosis virus). Pretreatment of RNA virus with halocyamine A reduced the infectivity of the virus toward host cells. The growth of marine bacteria, Achromobacter aquamarinus and Pseudomonas perfectomarinus, was also inhibited by halocyamine A but that of Alteromonas putrefaciens and Vibrio anguillarum was not. These results suggest that halocyamine may have a role in the defense mechanisms of H. roretzi against marine viruses and bacteria.

Animals↗
PubMed1990

Halocyamines: novel antimicrobial tetrapeptide-like substances isolated from the hemocytes of the solitary ascidian Halocynthia roretzi.

Two novel antimicrobial tetrapeptide-like substances, halocyamine A and B, were isolated from the solitary ascidian Halocynthia roretzi by a procedure including extraction steps, chromatographies on coarse and fine HP-20 columns, and preparative reversed-phase high-performance liquid chromatography. The structures of halocyamine A and B were determined to be L-histidyl-L-6,7-dihydroxyphenylalanylglycyl-6-bromo-8,9-didehy drotryptamine and L-threonyl-L-6,7-dihydroxyphenylalanyl-L-histidyl-6-bromo-8,9- didehydrotryptamine, respectively, by spectral analyses and degradation studies. Besides antimicrobial activities against several kinds of bacteria and yeasts, both of them showed cytotoxic activities against neuronal cells cultured from rat fetal brain, mouse neuroblastoma N-18 cells, and human hepatoma Hep-G2 cells. They were only detected in the "morula"-like cells, which are of the most abundant cell type among the hemocytes of H. roretzi.

Animals↗
PubMed1987

Filter transient response to EEG waveforms.

The response of two types of linear filters to sinusoidal bursts was calculated to demonstrate how filters can distort EEG waveforms. Results show that the wider the filter bandwidth the less is the distortion, and for a given bandwidth, the higher the filter order the greater the distortion. The response of a linear phase filter was also calculated to demonstrate that this type of filter can also cause waveform distortion, although it is normally less than that caused by Butterworth, Tchebychev and elliptic filters.

Electroencephalography↗