Publications by authors named "Reiko Nagata"

22 Publications

  • Page 1 of 1

Complete Genome Sequence of Mycobacterium avium subsp. Strain 42-13-1, Isolated in Japan.

Microbiol Resour Announc 2021 Apr 15;10(15). Epub 2021 Apr 15.

National Institute of Animal Health, National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan.

Here, we report the complete genome sequence of subsp. strain 42-13-1, isolated from cattle presenting with chronic diarrhea caused by Johne's disease in Japan, which was assembled via long- and short-read hybrid assembly.
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http://dx.doi.org/10.1128/MRA.00084-21DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8050961PMC
April 2021

The enhancement of Th1 immune response by anti-PD-L1 antibody in cattle infected with Mycobacterium avium subsp. paratuberculosis.

J Vet Med Sci 2021 Feb 7;83(2):162-166. Epub 2020 Dec 7.

Department of Disease Control, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060-0818, Japan.

Johne's disease, caused by Mycobacterium avium subsp. paratuberculosis (MAP), is a chronic enteritis of ruminants. Previous studies have shown that programmed death-ligand 1 (PD-L1) is associated with the disease progression, and PD-L1 blockade activates MAP-specific Th1 responses in vitro. Here, we performed anti-PD-L1 antibody administration using 2 MAP-infected cattle at the late subclinical stage of infection. After administration, bacterial shedding was reduced or maintained at a low level. Additionally, MAP-specific Th1 cytokine production was upregulated, and CD69 expression was increased in T cells. Collectively, the treatment has a potential as a novel control method against Johne's disease.
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http://dx.doi.org/10.1292/jvms.20-0590DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7972883PMC
February 2021

A Novel Real-Time PCR-Based Screening Test with Pooled Fecal Samples for Bovine Johne's Disease.

J Clin Microbiol 2020 11 18;58(12). Epub 2020 Nov 18.

Division of Bacterial and Parasitic Disease, National Institute of Animal Health, National Agriculture and Food Research Organization, Tsukuba, Japan.

Johne's disease (JD) is an economically important infectious disease in livestock farming caused by subsp. As an alternative to serological tests, which are used mainly for the screening of whole herds, we developed a novel ResoLight-based real-time PCR (RL-PCR) assay with pooled fecal samples for the detection of fecal shedders in cattle herds. The RL-PCR assay included an internal amplification control (IC) which was amplified using the same primer pair as the target molecule subsp. IS and differentiated based on melting temperatures. Individual fecal suspensions were pooled and concentrated by centrifugation to avoid a loss of sensitivity by the dilution effect. Combined with a DNA extraction kit (Johne-PureSpin; FASMAC), no inhibition of PCR amplification was observed with up to 15 fecal samples in a pool. The detection limit of RL-PCR at a pool size of 10 was 10 subsp. organisms per gram of feces, which was comparable to that of individual testing. A total of 2,654 animals in 12 infected herds were screened by individual antibody-enzyme-linked immunosorbent assay (ELISA) and the RL-PCR assay using pooled feces. Fifty animals were diagnosed with JD through the screening by RL-PCR, compared with only 5 by ELISA (which were also positive in RL-PCR). In 7 JD-free herds, the results of 4 out of 327 pools (1.2%) were invalid due to the lack of IC amplification, and then animals were confirmed negative individually. Our results suggest that implementation of herd screening by pooled RL-PCR would advance the monitoring and control of JD in cattle herds.
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http://dx.doi.org/10.1128/JCM.01761-20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7685889PMC
November 2020

Prostaglandin E Induction Suppresses the Th1 Immune Responses in Cattle with Johne's Disease.

Infect Immun 2018 05 23;86(5). Epub 2018 Apr 23.

Department of Disease Control, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Japan.

Johne's disease, caused by subsp. , is a bovine chronic infection that is endemic in Japan and many other countries. The expression of immunoinhibitory molecules is upregulated in cattle with Johne's disease, but the mechanism of immunosuppression is poorly understood. Prostaglandin E (PGE) is immunosuppressive in humans, but few veterinary data are available. In this study, functional and kinetic analyses of PGE were performed to investigate the immunosuppressive effect of PGE during Johne's disease. PGE treatment decreased T-cell proliferation and Th1 cytokine production and upregulated the expression of immunoinhibitory molecules such as interleukin-10 and programmed death ligand 1 (PD-L1) in peripheral blood mononuclear cells (PBMCs) from healthy cattle. PGE was upregulated in sera and intestinal lesions of cattle with Johne's disease. stimulation with Johnin purified protein derivative (J-PPD) induced cyclooxygenase-2 (COX-2) transcription, PGE production, and upregulation of PD-L1 and immunoinhibitory receptors in PBMCs from cattle infected with subsp. Therefore, Johnin-specific Th1 responses could be limited by the PGE pathway in cattle. In contrast, downregulation of PGE with a COX-2 inhibitor promoted J-PPD-stimulated CD8 T-cell proliferation and Th1 cytokine production in PBMCs from the experimentally infected cattle. PD-L1 blockade induced J-PPD-stimulated CD8 T-cell proliferation and interferon gamma production Combined treatment with a COX-2 inhibitor and anti-PD-L1 antibodies enhanced J-PPD-stimulated CD8 T-cell proliferation , suggesting that the blockade of both pathways is a potential therapeutic strategy to control Johne's disease. The effects of COX-2 inhibition warrant further study as a novel treatment of Johne's disease.
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http://dx.doi.org/10.1128/IAI.00910-17DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5913856PMC
May 2018

A case of rapidly-growing atypical polypoid adenomyoma which was histologically diagnosed before operation and removed by a laparoscopic resection.

Taiwan J Obstet Gynecol 2018 Feb;57(1):115-118

Department of Obstetrics and Gynecology, Tokyo Women's Medical University, Tokyo, Japan.

Objective: Atypical polypoid adenomyoma (APAM) is an epithelial-mesenchymal mixed tumor which often develops in the uterine cavity of reproductive age women, requiring preservation of the reproductive functions. Preoperative endometrial biopsy may not yield histological diagnosis as the tumor is a solid smooth muscle tumor. The standard treatment option is a hysteroscopic resection for the diagnosis and the treatment at the same time.

Case Report: We report a case of rapidly-growing APAM successfully diagnosed preoperatively via transcervical punch biopsy followed by a laparoscopic resection. The mass was relatively large, had been located in the lower segment of the uterus, and the area of contact with the muscular layers was large. It was a complete removal and no recurrence had been observed 9 months after the operation.

Conclusion: This is the first report of APAM treated by laparoscopic resection. The method may be a useful alternative when hysteroscopic surgery is inappropriate.
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http://dx.doi.org/10.1016/j.tjog.2017.12.019DOI Listing
February 2018

Evaluation of fecal shedding and antibody response in dairy cattle infected with paratuberculosis using national surveillance data in Japan.

Prev Vet Med 2018 Jan 11;149:38-46. Epub 2017 Nov 11.

Epidemiology Unit, Division of Viral Disease and Epidemiology, National Institute of Animal Health, National Agriculture and Food Research Organization, Kannondai 3-1-5, Tsukuba, Ibaraki, 305-0856, Japan.

Paratuberculosis or Johne's disease (JD), is a chronic infectious disease causing intractable diarrhea in cattle, which leads to less productivity, such as decreased milk yield, and lower daily weight gain. As a control measure against JD in cattle, national serological surveillance has been conducted in Japan since 1998. To conduct modeling studies that are useful to evaluate the effectiveness of control measures against JD, reliable parameter values, such as length of time from infection to the start of fecal shedding or antibody expression, are especially important. These parameters in the Japanese cattle population are assumed to be different from those in other countries with a higher prevalence of JD or in experimental infection settings; therefore, they must be estimated for the cattle population in Japan. Data from national surveillance conducted in Tokachi District, Hokkaido Prefecture, were used for this study. Using data from JD diagnostic tests for all cattle in Tokachi District between 1998 and 2014, all testing histories for infected animals were estimated as the number of tested cattle and positive cattle at each age of month for both fecal and antibody tests. A deterministic mathematical model for JD development, from infection to fecal shedding and antibody expression in infected cattle, was constructed to obtain the probability of testing positive when applied to both fecal and antibody tests at a given age. Likelihood was obtained from these estimated test results and best values for parameters were obtained using the Markov Chain Monte-Carlo method. Fifty-five percent of infected cattle were projected to have a transient shedding period, which was estimated to start 12 months after infection and last for 4 months. Persistent shedding was projected to occur in all infected cattle, and estimated to begin 7-84 months from infection. Following persistent shedding, antibody expression was estimated to start 7 months later. These values are useful for developing models to evaluate the status of JD infection and the effectiveness of control measures in the Japanese cattle population.
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http://dx.doi.org/10.1016/j.prevetmed.2017.10.009DOI Listing
January 2018

Systemic mycobacteriosis caused by 'Mycobacterium avium subspecies hominissuis' in a 14-month-old Japanese black beef steer.

J Vet Med Sci 2017 Aug 10;79(8):1384-1388. Epub 2017 Jul 10.

Pathology and Pathophysiology Research Division, National Institute of Animal Health, National Agriculture and Food Research Organization (NARO), 3-1-5 Kannondai, Tsukuba, Ibaraki 305-0856, Japan.

A 14-month-old Japanese black beef steer presented with severe chronic diarrhea and emaciation and was euthanized. Postmortem examination showed thickened and corrugated intestinal mucosa and enlarged granulomatous mesenteric lymph nodes with caseating necrosis. Numerous epithelioid cells and multinucleated giant cells infiltrated in the lamina propria and the submucosal tissue of the intestines. These cells were also observed in the systemic organs. Many acid-fast bacilli were detected in the cytoplasm of these cells and were identified as 'Mycobacterium avium subsp. hominissuis' (Mah) on the basis of the results of molecular examinations and immunohistochemistry. These findings indicate that Mah can cause systemic mycobacteriosis, and this unique infection needs to be distinguished from Johne's disease and tuberculosis in cattle.
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http://dx.doi.org/10.1292/jvms.17-0204DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5573826PMC
August 2017

Gorlin syndrome with an ovarian leiomyoma associated with a PTCH1 second hit.

Am J Med Genet A 2016 Apr 18;170A(4):1029-34. Epub 2016 Jan 18.

Institute of Medical Genetics, Tokyo Women's Medical University, Tokyo, Japan.

We describe a Gorlin syndrome (GS) case with two different second hit mutations of PTCH1, one in a keratocystic odontogenic tumor (KCOT) and the other in an ovarian leiomyoma. GS is a rare genetic condition manifesting as multiple basal cell nevi associated with other features such as medulloblastomas, skeletal abnormalities, and ovarian fibromas. A 21-year-old Japanese woman with a history of two KCOTs was diagnosed with GS according to clinical criteria. A PTCH1 mutation, c.1427del T, was detected in peripheral blood. A novel PTCH1 mutation, c.264_265insAATA, had been found in the maxillary KCOT as a second hit mutation. More recently, the ovarian tumor was detected during a gynecological examination. Laparoscopic adnexectomy was performed, and the pathological diagnosis of the ovarian tumor was leiomyoma. Interestingly, another novel mutation, loss of heterozygosity spanning from 9q22.32 to 9q31.2, including PTCH1 and 89 other genes, was detected in this ovarian tumor, providing evidence of a second hit mutation. This is the first report describing a GS-associated ovarian tumor carrying a second hit in the PTCH1 region. We anticipate that accumulation of more cases will clarify the importance of second hit mutations in ovarian tumor formation in GS.
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http://dx.doi.org/10.1002/ajmg.a.37517DOI Listing
April 2016

Bovine Immunoinhibitory Receptors Contribute to Suppression of Mycobacterium avium subsp. paratuberculosis-Specific T-Cell Responses.

Infect Immun 2016 01 19;84(1):77-89. Epub 2015 Oct 19.

Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan

Johne's disease (paratuberculosis) is a chronic enteritis in cattle that is caused by intracellular infection with Mycobacterium avium subsp. paratuberculosis. This infection is characterized by the functional exhaustion of T-cell responses to M. avium subsp. paratuberculosis antigens during late subclinical and clinical stages, presumably facilitating the persistence of this bacterium and the formation of clinical lesions. However, the mechanisms underlying T-cell exhaustion in Johne's disease are poorly understood. Thus, we performed expression and functional analyses of the immunoinhibitory molecules programmed death-1 (PD-1)/PD-ligand 1 (PD-L1) and lymphocyte activation gene 3 (LAG-3)/major histocompatibility complex class II (MHC-II) in M. avium subsp. paratuberculosis-infected cattle during the late subclinical stage. Flow cytometric analyses revealed the upregulation of PD-1 and LAG-3 in T cells in infected animals, which suffered progressive suppression of interferon gamma (IFN-γ) responses to the M. avium subsp. paratuberculosis antigen. In addition, PD-L1 and MHC-II were expressed on macrophages from infected animals, consistent with PD-1 and LAG-3 pathways contributing to the suppression of IFN-γ responses during the subclinical stages of M. avium subsp. paratuberculosis infection. Furthermore, dual blockade of PD-L1 and LAG-3 enhanced M. avium subsp. paratuberculosis-specific IFN-γ responses in blood from infected animals, and in vitro LAG-3 blockade enhanced IFN-γ production from M. avium subsp. paratuberculosis-specific CD4(+) and CD8(+) T cells. Taken together, the present data indicate that M. avium subsp. paratuberculosis-specific T-cell exhaustion is in part mediated by PD-1/PD-L1 and LAG-3/MHC-II interactions and that LAG-3 is a molecular target for the control of M. avium subsp. paratuberculosis-specific T-cell responses.
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http://dx.doi.org/10.1128/IAI.01014-15DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4694020PMC
January 2016

Detection and confirmation of Mycobacterium avium subsp. paratuberculosis in direct quantitative PCR positive fecal samples by the manual fluorescent MGIT culture system.

J Vet Med Sci 2014 Jan 20;76(1):65-72. Epub 2013 Sep 20.

Bacterial and Parasitic Disease Research Division, National Institute of Animal Health, National Agriculture and Food Research Organization, Tsukuba, Ibaraki 305-0856, Japan.

An efficient protocol for the manual fluorescent MGIT culture system combined with rapid confirmation of Mycobacterium avium subsp. paratuberculosis (MAP) growth in the broth culture was established and evaluated for the detection of viable MAP in direct quantitative PCR (QPCR) positive bovine feces. Manually detected fluorescence emissions from MGIT tubes were analyzed objectively using an open source software, ImageJ. For molecular confirmation of MAP growth, DNA samples harvested by simply boiling the broth, an inexpensive and time- and labor-saving DNA preparation method, yielded adequate results. The sheep strain of MAP required longer incubation time relative to the cattle strain, suggesting that the MGIT system may not support well the growth of ovine isolates as described previously. Of 61 direct QPCR positive bovine feces, the recovery rate of MAP in the MGIT system (62.3%) was significantly higher (P<0.05) than that using 7H10 agar-based slants (44.3%). The time to obtain a final result for fecal culture by the MGIT system was several weeks earlier compared to solid media. In MGIT culture positive samples, the time to detect fluorescence was correlated with the DNA quantity detected in fecal QPCR. As a positive result in the direct fecal QPCR test does not mean fecal excretion of viable MAP, bacterial isolation by fecal culture could be conducted to verify the QPCR result. For this purpose, the manual MGIT system is a sensitive and rapid culture method at least for bovine samples.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3979941PMC
http://dx.doi.org/10.1292/jvms.13-0366DOI Listing
January 2014

Use of enoyl coenzyme A hydratase of Mycobacterium avium subsp. paratuberculosis for the serological diagnosis of Johne's disease.

Vet Immunol Immunopathol 2013 Oct 5;155(4):253-8. Epub 2013 Jul 5.

Bacterial and Parasitic Disease Research Division, National Institute of Animal Health, NARO, 3-1-5, Kannondai, Tsukuba, Ibaraki 305-0856, Japan. Electronic address:

Johne's disease (JD), caused by Mycobacterium avium subspecies paratuberculosis (MAP), remains difficult to control because of the lack of specific and sensitive diagnostic tests. In order to improve the specificity of sero-diagnosis for JD, the phage display library derived from genomic DNA of MAP was immunoscreened to identify novel antigenic targets. We selected a clone using antibodies from MAP experimentally infected cattle, and annotated its coding sequence as MAP1197 in the MAP genome, which encoded "echA12_2" in the MAP protein (Map-echA) belonging to Enoyl-CoA hydratase, known as a crotonase enzyme. The Map-echA was expressed in Esherichia coli and purified as a histidine-tag recombinant protein (rMap-echA), and the diagnostic potential of the protein was further evaluated by enzyme-linked immunosorbent assays (ELISA). Antibody responses to rMap-echA were higher in MAP-infected cattle than in uninfected cattle. The specificity of the Map-echA ELISA was also confirmed by evaluation with hyper-immune sera against various kinds of Mycobacterium species. Furthermore, in all experimentally infected cattle the antibody against rMap-echA was detected 2-7months earlier than by a commercially available ELISA kit. These results suggested that Map-echA can be used as a specific and sensitive serological diagnostic antigen for the detection of MAP infection.
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http://dx.doi.org/10.1016/j.vetimm.2013.06.020DOI Listing
October 2013

Detection of antibody responses against Mycobacterium avium subsp. paratuberculosis stress-associated proteins within 30 weeks after infection in cattle.

Vet Immunol Immunopathol 2012 Nov 11;150(1-2):101-11. Epub 2012 Sep 11.

Bacterial and Parasitic Disease Research Division, National Institute of Animal Health, National Agriculture and Food Research Organization, 3-1-5 Kannondai, Tsukuba, Ibaraki 305-0856, Japan.

In this study, humoral immune responses in cattle against Mycobacterium avium subsp. paratuberculosis (MAP) stress-associated recombinant proteins were assessed longitudinally by ELISA during the first 30 weeks after MAP infection. A total of 11 MAP genes previously identified by proteomic analysis were selected for cloning and expression. These included possible general stress-associated proteins of MAP and proteins expressed in vivo in MAP-infected sheep at an early stage of infection. An increase in the antibody levels against 5 recombinant antigen preparations (MAP1027c, MAP1339, MAP1588c, MAP1589c and MAP2411) was seen in MAP-infected calves (n=16) but not in control calves (n=3) over the time examined. Antibody responses were recorded as early as two weeks post-inoculation, and 87.5% of the inoculated cattle responded to at least one of the five immunogenic antigen preparations within the first 30 weeks of infection, suggesting that these proteins identified in the in vitro models of stress were also expressed in vivo in MAP-infected cattle at a relatively early stage after infection and therefore stimulate the host's immune system. It has been assumed that the sensitivity of antibody ELISA tests is dependent on the stage of infection and the age of the animals. However, we have provided some evidence that humoral immunity occurs at an early stage of paratuberculosis and can be detected using appropriate antigens such as MAP stress-associated proteins.
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http://dx.doi.org/10.1016/j.vetimm.2012.09.003DOI Listing
November 2012

Decreased expression of matrix metalloproteinase-9 and increased expression of tissue inhibitors of matrix metalloproteinase-1 in paratuberculosis-infected cattle in the ELISA-negative subclinical stage.

Anim Biotechnol 2011 Jan;22(1):44-9

Research Team for Paratuberculosis, National Institute of Animal Health, Tsukuba, Japan.

We investigated the gene expression of matrix metalloproteinases-9 (MMP-9) and tissue inhibitors of matrix metalloproteinases-1 (TIMP-1) in peripheral blood cells from infected cattle with Mycobacterium avium subsp. paratuberculosis (Map) in the ELISA-negative subclinical stage compared with uninfected control cattle. Significant decreased MMP-9 expression and increased TIMP-1 expression were found in peripheral blood cells from Map-infected cattle after stimulation with Map lysate and Map purified protein derivative (PPD) than in control cattle by real-time RT-PCR analysis. In contrast to the uninfected controls, the activity of MMP-9 was also decreased in peripheral blood cell culture supernatants from Map-infected cattle at 24 hr after Map lysate and MapPPD stimulation by gelatin zymography analysis. As a result, the MMP-9 may play an important role in the development of Mycobacterium avium subsp. paratuberculosis disease.
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http://dx.doi.org/10.1080/10495398.2010.536096DOI Listing
January 2011

A specific induction of interleukin-10 by the Map41 recombinant PPE antigen of Mycobacterium avium subsp. paratuberculosis.

Vet Immunol Immunopathol 2010 May 13;135(1-2):71-78. Epub 2009 Nov 13.

Research Team for Paratuberculosis, National Institute of Animal Health, 3-1-5 Kannondai, Tsukuba, Ibaraki 305-0856, Japan.

Interleukin-10 (IL-10) is not only an essential immunoregulator in host immunity, but also it accounts for the intracellular survival of mycobacteria because of its inhibitory activity against anti-mycobacterial functions of macrophage. It has been also indicated that blood cells from calves infected with Mycobacterium avium subsp. paratuberculosis (Map) produce a large amount of IL-10 after stimulation with Map antigen, and it leads to suppression of Interferon-gamma (IFN-gamma) production in T-cells. This characteristic expression of IL-10 in Map-infected cattle seems to be playing important roles in the pathogenesis of Johne's disease caused by Map, and could be an important diagnostic indicator. The aim of this study was to investigate the diagnostic significance of IL-10 production from blood cells stimulated by a PPE (Proline-Proline-Glutamic acid) protein family of Map. The recombinant PPE protein, Map41, which has been reported as one of the IFN-gamma inducing antigens of Map, also strongly induced IL-10 from macrophages obtained from infected calves. The elicited IL-10 production in response to Map41 from experimentally infected calves was as early as 2 weeks after the inoculation of Map, and the IL-10 production was detected earlier than that of IFN-gamma. The blood cells from calves immunized with Map produced higher amounts of IL-10 against Map41 stimulation than those of calves immunized with various Mycobacterium species. Furthermore, this IL-10 induction also showed high specificity to Map in guinea pigs experimentally infected with various Mycobacterium species. These observations suggest that IL-10 assay is a useful diagnostic method in the early stage of Johne's disease.
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http://dx.doi.org/10.1016/j.vetimm.2009.11.002DOI Listing
May 2010

Preparation of an in-house reference material containing fumonisins in Thai rice and matrix extension of the analytical method for Japanese rice.

Toxins (Basel) 2009 12 8;1(2):188-95. Epub 2009 Dec 8.

Biotechnology Research Centre, Malaysian Agricultural Research and Development Institute, GPO, P.O. Box 12301, 50774 Kuala Lumpur, Malaysia.

Mycotoxin contamination in rice is less reported, compared to that in wheat or maize, however, some Fusarium fungi occasionally infect rice in the paddy field. Fumonisins are mycotoxins mainly produced by Fusarium verticillioides, which often ruins maize. Rice adherent fungus Gibberella fujikuroi is taxonomically near to F. verticillioides, and there are sporadic reports of fumonisin contamination in rice from Asia, Europe and the United States. Therefore, there exists the potential risk of fumonisin contamination in rice as well as the need for the validated analytical method for fumonisins in rice. Although both natural and spiked reference materials are available for some Fusarium mycotoxins in matrices of wheat and maize, there are no reference materials for Fusarium mycotoxins in rice. In this study, we have developed a method for the preparation of a reference material containing fumonisins in Thai rice. A ShakeMaster grinding machine was used for the preparation of a mixed material of blank Thai rice and F. verticillioides-infected Thai rice. The homogeneity of the mixed material was confirmed by one-way analysis of variance, which led this material to serve as an in-house reference material. Using this reference material, several procedures to extract fumonisins from Thai rice were compared. Accordingly, we proved the applicability of an effective extraction procedure for the determination of fumonisins in Japanese rice.
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http://dx.doi.org/10.3390/toxins1020188DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3202782PMC
December 2009

Limited surveillance of fumonisins in brown rice and wheat harvested in Japan.

J Food Prot 2009 Jun;72(6):1327-31

National Food Research Institute, National Agriculture and Food Research Organization, 2-1-12 Kannondai, Tsukuba 305-8642, Japan.

Fumonisins are mycotoxins mainly produced by Fusarium verticillioides, which is a major contaminant of corn. However, there are sporadic reports of fumonisin contamination in wheat worldwide. The rice adherent fungus Gibberella fujikuroi is taxonomically closely related to F. verticillioides. Therefore, the potential risk of fumonisin contamination in rice and wheat is significant. Previously, a sensitive detection method utilizing liquid chromatography with tandem electrospray mass spectrometry (LC-ESI-MS-MS) was developed for the determination of fumonisins in brown rice. In the present study, the incidence of fumonisins in brown rice and wheat harvested in Japan was investigated using LC-ESI-MS-MS. Forty-eight rice samples and 47 wheat samples were screened and analyzed for the major B-type fumonisins: fumonisin B1 (FB1) and fumonisin B2 (FB2). About 1 kg of rice or wheat seed was divided into three subsamples, and 10 g from each subsample was used for the analysis. The limits of detection were 0.012 and 0.011 mg/kg for FBt and FB2, respectively, in rice samples and 0.010 and 0.008 mg/kg for FB1 and FB2, respectively, in wheat samples. The mean (standard deviation) recoveries of FB1 spiked at 0.50 mg/kg into toxin-free rice and wheat samples were 77.6 (4.2)% and 84.5 (3.1)%, respectively. One of the wheat samples was positive for FBt with a value greater than the limit of detection,but no fumonisin was found in any of the rice samples. This is the first report of fumonisins detected in Japanese wheat.
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http://dx.doi.org/10.4315/0362-028x-72.6.1327DOI Listing
June 2009

IL-18 expression in pigs following infection with Mycoplasma hyopneumoniae.

J Interferon Cytokine Res 2006 Sep;26(9):637-44

Department of Immunology, National Institute of Animal Health, Tsukuba, Ibaraki, 305-0856, Japan.

Little is known about the detail of the immune response during infection of pigs with Mycoplasma hyopneumoniae (Mhp). To further understand this important porcine pathogen, we examined the interleukin-18 (IL- 18) response in experimentally infected piglets. We found that large amounts of IL-18 were produced in the bronchoalveolar lavage fluids (BALF) of pigs experimentally infected with Mhp. However, the concentration of interferon-gamma (IFN-gamma) in the same BALF was negatively correlated with that of IL-18. The antibody response against Mhp was found to be associated with the IL-18 concentration in the BALF. Immunohistochemical staining revealed that both IL-18 and IL-18 receptor alpha chain (IL-18Ralpha) were present in macrophages and plasma cells in the lungs of Mhp-infected pigs. Lung mononuclear cells isolated from pneumonic lesions secreted IL-18 and prostaglandin E(2) (PGE(2)) in vitro, and PGE(2) production was enhanced by stimulation with IL-18. These results indicate that IL-18 produced in the pig lung contributes to the development of innate and acquired immune responses against Mhp as a proinflammatory cytokine rather than as an IFN-gamma-inducing factor and may be involved in immunomodulation in pigs.
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http://dx.doi.org/10.1089/jir.2006.26.637DOI Listing
September 2006

Bovine IL-18 ELISA: detection of IL-18 in sera of pregnant cow and newborn calf, and in colostrum.

J Immunoassay Immunochem 2005 ;26(3):203-13

National Institute of Animal Health, 3-1-5, Kannondai, Tsukuba, Ibaraki 305-0856, Japan.

In this study, we examined the concentration of bovine IL-18 in the sera of pregnant cows, their fetuses and newborn calves, and in colostrum in order to examine the role of IL-18 in bovine pregnancy and the neonatal period. A sandwich-ELISA to quantify bovine IL-18 was established using anti-porcine IL-18 monoclonal antibodies, which cross-reacted with bovine IL-18, and used it to measure the concentration of bovine IL-18 in the sera of pregnant cows, their fetuses and newborn calves, and in colostrum. Significant levels of IL-18 were detected in the sera of pregnant cows, but not in the sera obtained from the corresponding fetuses, umbilical arteries and veins. After birth, IL-18 levels in the sera of 1-day and 1-week old calves were low, and significantly increased in the sera of 1-month and 4-month old calves. IL-18 was also detected in colostrum, with the concentration of IL-18 in the first colostrum produced after delivery being the highest, and then decreasing depending on the number of milkings. Furthermore, the serum IL-18 concentration of newborn calves was increased after the oral administration of colostrum. These results suggest that IL-18 during bovine pregnancy and in the newborn period may play important roles in the maintenance of pregnancy and in the maturation of neonatal immunity.
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http://dx.doi.org/10.1081/IAS-200062487DOI Listing
September 2005

Expression cloning of gamma interferon-inducing antigens of Mycobacterium avium subsp. paratuberculosis.

Infect Immun 2005 Jun;73(6):3778-82

Immune System Section, Department of Immunology, National Institute of Animal Health, 3-1-5 Kannondai, Tsukuba, Ibaraki, Japan 305-0856.

Three recombinant proteins, Map10, Map39, and Map41, produced based on nucleotide sequences obtained from the screening of Mycobacterium avium subsp. paratuberculosis genomic library expressed in Escherichia coli significantly elicited gamma interferon production in peripheral blood mononuclear cells from infected cattle. Two of these proteins were members of the PPE protein family.
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http://dx.doi.org/10.1128/IAI.73.6.3778-3782.2005DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1111817PMC
June 2005

Generation of multinucleated giant cells in vitro from bovine monocytes and macrophages.

J Vet Med Sci 2004 Sep;66(9):1065-9

National Institute of Animal Health, Tsukuba, Japan.

The generation of multinucleated giant cells (MGC) from cells of the bovine monocyte-macrophage lineage was investigated. Freshly isolated monocytes were incubated with the conditioned medium (CM) of peripheral blood mononuclear cell cultures treated with Concanavalin A for 1-4 days (CM1 to CM4). Only CM1 generated MGC despite similar concentrations of IFNgamma in all CMs. Nevertheless, MGC formation from monocytes was enhanced by adding either macrophage colony-stimulating factor (M-CSF) or granulocyte-macrophage colony-stimulating factor (GM-CSF), MGC formations from macrophages were observed only when macrophages were cultured with GM-CSF plus CM. These results indicate that several mechanisms to generate MGC from bovine monocytes-macrophage lineage cells exist, and that GM-CSF is a major mediator of MGC formation in cattle.
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http://dx.doi.org/10.1292/jvms.66.1065DOI Listing
September 2004

Neutralization of interleukin-10 significantly enhances gamma interferon expression in peripheral blood by stimulation with Johnin purified protein derivative and by infection with Mycobacterium avium subsp. paratuberculosis in experimentally infected cattle with paratuberculosis.

Infect Immun 2004 Apr;72(4):2425-8

Paratuberculosis and Inflammatory Bowel Disease Research Team. Immune System Section, National Institute of Animal Health. Molecular Immunology Laboratory, National Institute of Agrobiological Sciences, 3-1-5 Kan-nondai, Tsukuba 305-0856, Japan.

Monoclonal antibody neutralization of interleukin-10 (IL-10) increased Johnin purified protein derivative-induced whole-blood gamma interferon (IFN-gamma) secretion 23-fold and also increased IFN-gamma secretion ninefold following in vitro Mycobacterium avium subsp. paratuberculosis infection of peripheral blood mononuclear cells. These results demonstrate the suppressive effect of IL-10 on immune responses to M. avium subsp. paratuberculosis infection in cattle.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC375198PMC
http://dx.doi.org/10.1128/IAI.72.4.2425-2428.2004DOI Listing
April 2004

A new synthetic approach to cyclic bis(3'-->5')diguanylic acid.

Nucleic Acids Res Suppl 2003 (3):103-4

Graduate School of Human Informatics, Nagoya University, Chikusa, Nagoya 464-8601, Japan.

We developed a novel synthesis of biologically important cyclic bis(3'-->5')diguanylic acid (cGpGp). The present synthesis includes two strategies different from those employed in an existing synthesis. They are the phosphoramidite method for the preparation of a guanylyl(3'-->5')guanylic acid intermediate and allyl protection for guanine bases and internucleotide linkages. These distinctive strategies have allowed the new synthesis to provide the target compound in a higher yield than that of the existing synthesis.
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http://dx.doi.org/10.1093/nass/3.1.103DOI Listing
October 2003