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M Onuma

Publications and source records attributed to M Onuma.

At least 73 records · Page 4Linked to original sources
PubMed1999

Characterization of proteolytic enzymes expressed in the midgut of Haemaphysalis longicornis.

The proteolytic activities present in midguts of both fed and unfed Haemaphysalis longicornis were assessed by using the gelatin-substrate gel electrophoresis and inhibitor sensitivity analyses. Three predominant (116, 48 and 48 kDa) and two weak (55 and 60 kDa) proteinase bands were commonly expressed in both unfed and fed ticks, while a weak 80 kDa band was only present in fed ticks. Consistent with observations on other tick species, proteolytic activity against the gelatin substrate was observed only under acidic conditions. Inhibition studies against the gelatin substrate using a panel of inhibitors showed that the predominant proteolytic enzymes of 40 and 48 kDa molecular mass are cysteine proteinases. These results are discussed in the context of host vaccination as an alternative tick control method to the current use of chemical acaricides.

Animals↗
PubMed1998

Sequence analysis of the major piroplasm surface protein gene of benign bovine Theileria parasites in east Asia.

Relatively benign Theileria parasites are widespread among cattle in East Asia. Although the parasites are presumed to be of the Theileria sergenti/Theileria buffeli/Theileria orientalis group, their taxonomic status and epidemiology have not been well defined. In the present study, theilerial DNA samples were collected from various East Asian countries, including Japan, Korea, Taiwan, and China. DNA sequences encoding a major piroplasm surface protein were amplified by polymerase chain reaction, followed by cloning into a plasmid vector. More than 20 DNA clones derived from parasite DNA of a single infected animal were examined for their restriction-fragment-length polymorphism, showing that they were classified into four major types. Sequence analysis revealed six types of DNA sequences encoding major piroplasm surface protein with homologies of between 75 and 91%. Of the six sequences, four were identical to those previously reported, while the other two appeared to be new sequences. Among the DNA clones derived from a single infected animal, two to three distinct sequences were often found. Phylogenetic analysis of the six major piroplasm surface protein sequences indicates that five of the six are closely related to each other, and that all are distantly related to the homologous genes of Theileria annulata and Theileria parva. The results suggest that, in addition to those described as T. sergenti/T. buffeli/T. orientalis, there may be some undefined Theileria species distributed in East Asia, and that many cattle are infected with mixed populations of geographically variable Theileria parasites.

Amino Acid Sequence↗
PubMed1998

Phylogeny of benign Theileria species from cattle in Thailand, China and the U.S.A. based on the major piroplasm surface protein and small subunit ribosomal RNA genes.

The major piroplasm surface protein and small subunit ribosomal RNA genes of benign Theileria species isolated from cattle in China, Thailand and the U.S.A. were amplified by polymerase chain reaction, cloned and sequenced. The major piroplasm surface protein genes of these three isolates were more than 89% identical at amino-acid level. Several deletions in the gene from the Thai isolate led to considerable structural change through frame shifts of the major piroplasm surface protein. Phylogenetic analyses based on both of the major piroplasm surface protein and small subunit ribosomal RNA genes suggest that there may be a second cosmopolitan benign Theileria species infecting cattle in addition to the Theileria sergenti/buffeli/orientalis complex.

Amino Acid Sequence↗
PubMed1998

Theileria parasite infection in East Asia and control of the disease.

Bovine piroplasmosis caused by Theileria sergenti is a major cause of economic loss in grazing cattle in Japan. We found that parasite stocks and isolates consist of genetically and antigenically mixed populations. To differentiate among parasite populations bearing 3 allelic forms of p32/34, an immunodominant piroplasmin surface protein, 3 sets of oligonucleotide primers were designed to amplify either of 3 alleles of T. sergenti/T. buffeli/T. orientalis by polymerase chain reaction (PCR). By using this allele-specific PCR, we found that in East Asia the majority of bovines infected with benign Theileria parasites harbored mixed parasite population. As a possible means of controlling Theileria sergenti infection, we produced 2 candidate vaccines; a bactilovirus expressed recombinant p32 and a synthetic peptide containing of Lys-Glu Lys (KEK) motif. Immunization with either of two candidates resulted in lower parasitemia and reduced the severity of clinical symptoms as compared to control calves.

Alleles↗
PubMed1998

Antitumor effect of diphtheria toxin A-chain gene-containing cationic liposomes conjugated with monoclonal antibody directed to tumor-associated antigen of bovine leukemia cells.

Monoclonal antibody c143 against tumor-associated antigen (TAA) expressed on bovine leukemia cells was conjugated to cationic liposomes carrying a plasmid pLTR-DT which contained a gene for diphtheria toxin A-chain (DT-A) under the control of the long terminal repeat (LTR) of bovine leukemia virus (BLV) in the multicloning site of pUC-18. The specificity and antitumor effects of the conjugates were examined in vitro and in vivo using TAA-positive bovine B-cell lymphoma line as the target tumor. In vitro studies with the TAA-positive cell line indicated that luciferase gene-containing cationic liposomes associated with the c143 anti-TAA monoclonal antibody caused about 2-fold increase in luciferase activity compared with cationic liposomes having no antibody, and also that the c143-conjugated cationic liposomes containing pLTR-DT exerted selective growth-inhibitory effects on the TAA-positive B-cell line. Three injections of pLTR-DT-containing cationic liposomes coupled with c143 into tumor-bearing nude mice resulted in significant inhibition of the tumor growth. The antitumor potency of the c143-conjugated cationic liposomes containing pLTR-DT was far greater than that of normal mouse IgG-coupled cationic liposomes containing pLTR-DT as assessed in terms of tumor size. These results suggest that cationic liposomes bearing c143 are an efficient transfection reagent for BLV-infected B-cells lymphoma cells, and that the delivery of the pLTR-DT gene into BLV-infected B-cells by the use of such liposomes may become a useful technique for gene therapy of bovine leukosis.

Animals↗
PubMed1998

Non-antigenic and low allergic gelatin produced by specific digestion with an enzyme-coupled matrix.

Porcine gelatin (heat-denatured collagen) was digested with a bioreactor using an enzyme-coupled matrix (ECM) with purified collagenase. The digested gelatin, FreAlagin type R (M.W. range 200-10000 Da), was further purified by an HPLC system depending upon molecular size. The molecular weight range of the purified fractions, FreAlagin type P and type AD, were 200-500 and 2000-10000 Da, respectively, and glycine was the N-terminal amino acid of both types (> or =93%). ECM has the capability of digesting gelatin at a specific point in the sequence before glycine, and it was determined that FreAlagin type P consists of a tri-peptide fraction with the amino acid sequence Gly-X-Y. No types of FreAlagin exhibited any reactivity with gelatin-specific IgG antibody raised in guinea pigs, and they also possessed an extremely low reactivity with gelatin-specific IgE antibody from the sera of patients who had experienced an anaphylactic reaction against gelatin after vaccination or after eating gelatin-containing foods. From these results, it was determined that FreAlagin types R and AD were non-antigenic, low-allergic gelatins. FreAlagin type R, and especially type AD, had strong adsorption-blocking activity comparable to the level of bovine serum albumin, whereas type P and glycine had virtually no adsorption-blocking activity. Therefore, the new types of gelatin, FreAlagin types R and AD, are suitable for pharmaceutical use to avoid gelatin allergy.

Adsorption↗
PubMed1998

Pathogenesis of Marek's disease (MD) and possible mechanisms of immunity induced by MD vaccine.

Marek's disease (MD) is a lymphoproliferative disease of chicken, which is characterized by malignant T cell-lymphoma formation. This disease can be effectively prevented by vaccination with attenuated MD virus (MDV), apathogenic MDV or herpesvirus of turkey. MD vaccines are ones of a few vaccines which can prevent virus-induced tumor among mammalian and avian species. To determine the roles of T cell subsets in the protection mechanism, chickens vaccinated with an attenuated MDV (CVI988) were depleted of either CD4+ or CD8+ T cells by neonatal thymectomy and injections of monoclonal antibodies against chicken CD4 or CD8 molecules and then challenged with an oncogenic MDV. These birds were effectively protected from MDV-induced tumors. However, virus titers in CD4+ T cells, which are the main target cells for MDV-latent infection and subsequent transformation, were much higher in CD8-deficient vaccinated chickens than in untreated vaccinated chickens at the early stage of the latent phase. These results suggested that CD8+ T cell responses induced by the MD vaccine are essential for anti-virus but not anti-tumor effects. Here, we will discuss how the attenuated vaccine prevents chickens from lymphoma-formation by an oncogenic MDV.

Animals↗
PubMed1998

Pathogenesis of Babesia caballi infection in experimental horses.

The present study was designed to investigate the role of cytokines in the pathogenesis of Babesia caballi in experimentally infected horses. The expression of cytokine mRNA was determined by using reverse transcription-polymerase chain reaction in two B. caballi-infected horses for 2 weeks after the infection. In one horse, there was up-regulation of interferon-gamma, tumor necrosis factor-alpha (TNF-alpha) and interleukin-2 mRNAs, while in the second horse, expression of only TNF-alpha mRNA was up-regulated. No change was observed in interleukin-4 mRNA in both of the horses. To know the relation between nitric oxide (NO) production and pathogenesis, NO production was assayed in three dexamethasone treated-B. caballi-infected horses. Production of NO in all 3 horses increased significantly before death, although the parasitemia level remained very low. Treatment with NO inhibitor resulted in the suppression of NO production and increased parasitemia level in a horse, which died of the infection. The pathological examination showed that the main cause of the death was dyspnoea and pulmonary edema. Histopathologically, diffuse global mesangial proliferative glomerulonephritis was also observed. These results suggested that NO may be a critical effector molecule of immune defense against parasite. TNF-alpha and NO might be contributing to the pathogenesis in B. caballi infection.

Animals↗
PubMed1998

Seroprevalence and field isolation of bovine immunodeficiency virus.

A seroprevalence study of bovine lentivirus, known as bovine immunodeficiency virus (BIV), was conducted in 12 different dairy herds in Hokkaido, where some herds were a high prevalence of bovine leukemia virus (BLV) infection. Amongst 611 cattle, 28.6% of cattle were BLV-seropositive, and 11.7% of cattle were seropositive for BIV, while 4.2% of cattle were seropositive for both BIV and BLV. For the isolation of BIV, 19 samples of peripheral blood mononuclear cells (PBMC) and one sample of milk-derived leukocytes were prepared from BIV-seropositive cows. These PBMC and leukocyte preparations were then co-cultivated with cc81 cells, a cat cell line transformed by mouse sarcoma virus. BIV was isolated from 17 PBMC and one milk-derived leukocyte samples. The isolated viruses showed slow replication and syncytia formation. Major core antigen, p26 from these isolates were reacted with anti-BIV (American isolate R-29) serum. In addition, proviral DNA was detected in blood and milk samples by nested polymerase chain reaction and subsequent Southern blot hybridization. Nucleotide sequence analysis of the amplified pol gene products showed its 99.0 to 99.7% homology to that of BIV R-29. These results indicate that the Japanese BIV isolates appear to be antigenically and genetically similar to the American R-29. Since BIV was isolated from milk samples, BIV could possibly be transmitted through milk. This is the first report of BIV isolation in Japan.

Amino Acid Sequence↗
PubMed1998

Metastatic intracavitary cardiac aortic body tumor in a dog.

A mobile right-ventricular mass dynamically occluding the right ostium atrioventriculare in the systolic phase was detected in a 3-year-old male Tosa dog by echocardiography. At necropsy, multiple tumor masses of various sizes were observed in the heart base right ventricular lumen, myocardium, lung and liver. Dysplasia of tricuspid valve characterized by irregular shape of leaflets, upward malposition of large papillary muscles, and shortened and stout chordae tendineae was also detected. Histopathologically, the tumor cells, arranged in sheets or nests, were polyhedral with lightly eosinophilic and finely granular cytoplasm, and contained a hyperchromatic round or oval nucleus. By Grimelius' silver stain, tumor cells had cytoplasmic positive granules. Ultrastructurally, tumor cells contained characteristic small membrane-limited granules. This is the first report of metastatic intracavitary cardiac aortic body tumor in a dog.

Animals↗
PubMed1998

Analysis of immunodominant piroplasm surface protein genes of benign Theileria parasites distributed in China and Korea by allele-specific polymerase chain reaction.

Benign Theileria species distributed in China and Korea were characterized by allele-specific polymerase chain reaction (PCR), based on the sequences of major immunodominant piroplasm surface protein genes. In China, all the isolates contained Chitose (C) type parasites. One out of 5 isolates tested was a mixed population of Ikeda (I), C and B-2 types, whereas, all the isolates from Korea consisted of I type parasites. Except for 4 isolates, 29 isolates from Korea consisted of more than two types of parasites. The present data showed that benign Theileria species distributed in these countries were mixed parasite populations.

Alleles↗
PubMed1998

Survey of Theileria parasite infection in cattle in Taiwan.

An survey of Theileria parasite infection in cattle in Taiwan was carried out by polymerase chain reaction (PCR). A total of 491 blood samples, 105 from southern area and 386 from northern area, were collected from bovine in 16 different farms. From northern area, Theileria piroplasms could be seen in only 4 of 105 blood samples microscopically. However, when p32/34 genes (encoding immunodominant piroplasm surface proteins) were amplified by PCR, 15 blood samples were detected positive. They were analyzed by using allele-specific primers of 3 allelic forms of p32/34 and all contained C type of T. sergenti. Four blood samples were found infected with both C and B (T. buffeli) type parasites. Examination of 386 blood samples from southern area of Taiwan did not reveal any Theileria parasite microscopically, as well as by PCR amplification.

Animals↗
PubMed1998

Partial protection of severe combined immunodeficient mice against infection with Babesia microti by in vitro-generated CD4+ T cell clones.

Because CD4+ T cells were considered to be involved in protection against infection with Babesia microti, specific CD4+ T cells were generated in vitro from recovered BALB/c mice and their protective activity was tested in vivo. The cells produced varying amounts of interferon (IFN)-gamma in vitro in response to parasite antigen. In passive transfer experiments, three out of eleven T cell clones tested exerted protective activity in the early phase after infection. However, there seemed to be no correlation between this protection and in vitro IFN-gamma production by the T cell clones. Although the protection was partial and short-lived, the result provided direct evidence that CD4+ T cells play a crucial role in defense against B. microti.

Adoptive Transfer↗
PubMed1998

Identification of B cell epitopes of a 30 kDa Babesia equi merozoite surface protein.

A 30 kDa immunodominant surface antigen (p30) of Babesia equi has been used as a diagnostic antigen. The B cell epitopes on this molecule recognized by horse sera and monoclonal antibody (MAb) against p30, 36/133.97, were determined. A synthetic peptide of p30 with amino acid sequence of 123FYQEVLFKGFEAV135 exhibited strong positive reaction with the infected horse sera. In contrast, MAb 36/133.97 recognized different region of p30, as peptide synthesized with amino acid sequence of 27ASGAVVDFQLESI39 reacted strongly. In competitive inhibition ELISA, the binding of MAb 36/133.97 to recombinant p30 was inhibited by horse antibodies, although they did not recognize same or an overlapping epitope. The data on B cell epitopes in this study may be important in improving serodiagnostic methods of B. equi infection.

Amino Acid Sequence↗
PubMed1998

Epitope mapping of bovine leukemia virus transactivator protein Tax.

The immunogenicity of the bovine leukemia virus (BLV) transactivator protein (tax) was studied by mapping its B-cell and T-cell epitopes. Peptides (18 to 20-mer) overlapping by 10 amino acids, spanning whole amino acid sequence of BLVtax were synthesized. Recombinant BLVtax protein was used to immunize two different strains of mice, C57BL/6 and BALB/c. B-cell and T-cell epitopes of recombinant BLVtax protein was determined by screening all the 30 synthetic peptides, against immune serum in ELISA for antibody reactivity, and against immune spleen cells in lymphocyte proliferation assay for T-cell stimulation. Peptides with amino acids at position 111-130 and 131-150 were T-cell epitopes for C57BL/6 and BALB/c mice immune cells, respectively. B-cell epitope was mapped to amino acid sequence at 261-280 in both strains of mice. These results imply that BLVtax protein contains some of BLV- immunodominant epitopes and this information may be applied for designing an effective peptide vaccine capable of inducing neutralizing antibodies as well as cellular immunity.

Amino Acid Sequence↗
PubMed1998

Differentiation and quantification of Theileria sergenti piroplasm types using type-specific monoclonal antibodies.

Antigenic properties of two representative allelic products of the major piroplasm surface protein (MPSP) of Theileria sergenti were studied. Sera from cattle infected with either of Ikeda and Chitose types of the parasite reacted strongly with homologous but weakly with heterologous recombinant antigens in immunoblotting. Monoclonal antibodies (MoAbs) produced against the both allelic products of MPSP parasites reacted only to the immunizing antigen. These results suggested that crossreactivity between two allelic products is very low inspite of relatively high homology in their amino acid sequences. Double staining of parasitized erythrocyte smear using type-specific MoAbs by an indirect immunofluorescent assay revealed that the set of MoAbs was useful for quantitative and differential detection of each type of parasite in mixed population.

Animals↗
PubMed1998

Cytology of feather pulp lesions from Marek's disease (MD) virus-infected chickens and its application for diagnosis and prediction of MD.

Cytological changes of feather pulp lesions (FPL) sampled chronologically from the same specific-pathogen free chickens inoculated with Marek's disease virus serotype 1 (MDV) were examined, comparing with their histological changes. The birds having Marek's disease (MD) lymphomas or nerve lesions exhibited the characteristic lesion changes on the cytological smears of FPL; the initial non-suppurative inflammatory to the late lymphomatous FPL. The birds having neither the MD visceral lymphomas nor the nerve lesions manifested only non-suppurative inflammatory FPL on the cytological smears throughout the experimental periods. Histological evaluation of FPL sampled from the same birds confirmed as above mentioned cytological results. From these results, the cytological evaluation of FPL proved to be an effective diagnostic and prognostic tool in foreseeing MD incidence.

Animals↗