The Experts below are selected from a list of 225 Experts worldwide ranked by ideXlab platform
Yoshitane Dohi - One of the best experts on this subject based on the ideXlab platform.
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Growth inhibition of tumour cells by a liposome-encapsulated, mycolic Acid-containing glycolipid, trehalose 2,3,6'-trimycolate.
Immunology, 1991Co-Authors: Y Ohtsubo, M Furukawa, T Imagawa, N. Sugimoto, M Ikutoh, S Nakatsugi, Y Katoh, S Shinka, Yoshitane DohiAbstract:In vivo growth of syngeneic tumour cells in the peritoneal cavity was strongly inhibited by intraperitoneal injection of a liposome-encapsulated, mycolic Acid-containing glycolipid, trehalose 2,3,6'-trimycolate (TTM), derived from a non-pathogenic, Acid-Fast Bacterium. Gordona aurantiaca. Peritoneal macrophages from mice after this treatment lysed tumour cells in vitro at a low effector/target ratio, and their culture supernatant inhibited tumour cell growth. The supernatant inhibited growth of not only tumour necrosis factor (TNF)-sensitive tumour cells, but also TNF-insensitive tumour cells. This inhibitory activity was enhanced by addition of lipopolysaccharide (LPS) to the culture medium of the macrophages. The macrophages released more superoxide (O2-), TNF and interleukin-1 (IL-1) on LPS triggering, and the releases of these compounds were further increased by addition of recombinant interferon-gamma (IFN-gamma) to the medium. Moreover, splenic T cells of TTM liposome-primed mice were found to produce eight times more IFN-gamma upon stimulation with LPS. These results indicated that priming with TTM liposomes resulted in strong activation of macrophages, which lysed tumour cells directly and also inhibited tumour cell growth by released factors.
Nader Mosavari - One of the best experts on this subject based on the ideXlab platform.
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Production of MPT-64 recombinant protein from virulent strain of MycoBacterium bovis
Iranian journal of veterinary research, 2018Co-Authors: Maryam Mohammadi Tashakkori, Majid Tebianian, Mohammad Tabatabaei, Nader MosavariAbstract:Tuberculosis (TB) is a zoonotic infectious disease common to humans and animals which has been caused by a rod shaped, Acid Fast Bacterium, called MycoBacterium bovis. The rapid and sensitive detection is a great challenge for TB diagnosis. The virulent strains of MycoBacterium tuberculosis complex (MTBC) have 16 different regions of difference (RD) in their genome which encode some important antigens. The major protein of M. bovis 64 (MPT-64) is one of the main immune-stimulating antigens which are encode by RD-2 region. The aim of the present study was cloning, expression and purification of MPT-64 as a protein antigen of M. bovis in a prokaryotic system for the usage in the future diagnostic studies. In this experimental study, the mpt-64 gene with 687 bp has been proliferated from M. bovis whole genome by polymerase chain reaction (PCR) method. The PCR product has been digested by BamHI and HindIII restriction enzymes and cloned into pQE-30 plasmid. The recombinant protein has been expressed in the Escherichia coli M15 with induction by isopropyl-β-D-thiogalactopyranoside (IPTG). The expressed protein was analyzed on SDS-PAGE, and purified with Nitrilotriacetic Acid (Ni-NTA) column. Finally, its biological properties were confirmed in Western blotting method using specific antibodies. Data showed the successful cloning of mpt-64 gene (as a 687 bp segment) in expression vector. The MPT-64 recombinant protein was ideally expressed and purified as a 24 kDa protein. The result of this study indicated that MPT-64 recombinant protein (24 kDa) has been successfully expressed and purified in a prokaryotic system, so this protein could be used for differential diagnosis of pathogenic and non-pathogenic MycoBacterium, in suspected BTB cases.
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Cloning, expression, and purification of recombinant protein MPT-64 from a virulent strain of MycoBacterium bovis in a prokaryotic system.
International journal of mycobacteriology, 2016Co-Authors: Maryam Mohammadi Tashakkori, Majid Tebianian, Mohammad Tabatabaei, Nader MosavariAbstract:Tuberculosis (TB) is a zoonotic infectious disease common to humans and animals that is caused by the rod-shaped Acid-Fast Bacterium MycoBacterium bovis. Rapid and sensitive detection of TB is promoted by specific antigens. Virulent strains of the TB complex from M. bovis contain 16 regions of difference (RD) in their genome that encode important proteins, including major protein of MycoBacterium Tuberculosis 64 (MBT-64, which is a primary immune-stimulating antigen encoded by RD-2. In this study, we cloned, expressed, and purified MPT-64 as a potent M. bovis antigen in a prokaryotic system for use in future diagnostic studies. The antigenic region of the Mpt64 gene was investigated by bioinformatics methods, cloned into the PQE-30 plasmid, and expressed in Escherichia coli M15 cells, followed by isopropyl β-d-1-thiogalactopyranoside induction. The expressed protein was analyzed sodium dodecyl sulfate polyacrylamide gel electrophoresis and purified using a nickel-affinity column. Biological activity was confirmed by western blot using specific antibodies. Our data verified the successful cloning of the Mpt64 gene (687-bp segment) via the expression vector and purification of recombinant MPT-64 as a 24-kDa protein. These results indicated successful expression and purification of recombinant MPT-64 protein in a prokaryotic system. This protein can be used for serological diagnosis, improved detection of pathogenicity and non-pathogenicity between infected cattle, and for verification of suspected cases of bovine TB. Copyright © 2016.
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Cloning, expression, and purification of recombinant protein MPT-64 from a virulent strain of MycoBacterium bovis in a prokaryotic system.
International Journal of Mycobacteriology, 2016Co-Authors: Maryam Mohammadi Tashakkori, Majid Tebianian, Mohammad Tabatabaei, Nader MosavariAbstract:Abstract Objective Tuberculosis (TB) is a zoonotic infectious disease common to humans and animals that is caused by the rod-shaped Acid-Fast Bacterium MycoBacterium bovis. Rapid and sensitive detection of TB is promoted by specific antigens. Virulent strains of the TB complex from M. bovis contain 16 regions of difference (RD) in their genome that encode important proteins, including major protein of MycoBacterium Tuberculosis 64 (MBT-64, which is a primary immune-stimulating antigen encoded by RD-2. In this study, we cloned, expressed, and purified MPT-64 as a potent M. bovis antigen in a prokaryotic system for use in future diagnostic studies. Methods The antigenic region of the Mpt64 gene was investigated by bioinformatics methods, cloned into the PQE-30 plasmid, and expressed in Escherichia coli M15 cells, followed by isopropyl β- d -1-thiogalactopyranoside induction. The expressed protein was analyzed sodium dodecyl sulfate polyacrylamide gel electrophoresis and purified using a nickel-affinity column. Biological activity was confirmed by western blot using specific antibodies. Results Our data verified the successful cloning of the Mpt64 gene (687-bp segment) via the expression vector and purification of recombinant MPT-64 as a 24-kDa protein. Conclusion These results indicated successful expression and purification of recombinant MPT-64 protein in a prokaryotic system. This protein can be used for serological diagnosis, improved detection of pathogenicity and non-pathogenicity between infected cattle, and for verification of suspected cases of bovine TB.
Y Ohtsubo - One of the best experts on this subject based on the ideXlab platform.
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Growth inhibition of tumour cells by a liposome-encapsulated, mycolic Acid-containing glycolipid, trehalose 2,3,6'-trimycolate.
Immunology, 1991Co-Authors: Y Ohtsubo, M Furukawa, T Imagawa, N. Sugimoto, M Ikutoh, S Nakatsugi, Y Katoh, S Shinka, Yoshitane DohiAbstract:In vivo growth of syngeneic tumour cells in the peritoneal cavity was strongly inhibited by intraperitoneal injection of a liposome-encapsulated, mycolic Acid-containing glycolipid, trehalose 2,3,6'-trimycolate (TTM), derived from a non-pathogenic, Acid-Fast Bacterium. Gordona aurantiaca. Peritoneal macrophages from mice after this treatment lysed tumour cells in vitro at a low effector/target ratio, and their culture supernatant inhibited tumour cell growth. The supernatant inhibited growth of not only tumour necrosis factor (TNF)-sensitive tumour cells, but also TNF-insensitive tumour cells. This inhibitory activity was enhanced by addition of lipopolysaccharide (LPS) to the culture medium of the macrophages. The macrophages released more superoxide (O2-), TNF and interleukin-1 (IL-1) on LPS triggering, and the releases of these compounds were further increased by addition of recombinant interferon-gamma (IFN-gamma) to the medium. Moreover, splenic T cells of TTM liposome-primed mice were found to produce eight times more IFN-gamma upon stimulation with LPS. These results indicated that priming with TTM liposomes resulted in strong activation of macrophages, which lysed tumour cells directly and also inhibited tumour cell growth by released factors.
Joseph O Falkinham - One of the best experts on this subject based on the ideXlab platform.
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fluorescent Acid Fast microscopy for measuring phagocytosis of mycoBacterium avium mycoBacterium intracellulare and mycoBacterium scrofulaceum by tetrahymena pyriformis and their intracellular growth
Applied and Environmental Microbiology, 2001Co-Authors: Eileen D Strahl, Glenda E Gillaspy, Joseph O FalkinhamAbstract:Fluorescent Acid-Fast microscopy (FAM) was used to enumerate intracellular MycoBacterium avium, MycoBacterium intracellulare, and MycoBacterium scrofulaceum in the ciliated phagocytic protozoan Tetrahymena pyriformis. There was a linear relationship between FAM and colony counts of M. avium cells both from cultures and within protozoa. The Ziehl-Neelsen Acid-Fast stain could not be used to enumerate intracellular mycobacteria because uninfected protozoa contained Acid-Fast, Bacterium-like particles. Starved, 7-day-old cultures of T. pyriformis transferred into fresh medium readily phagocytized M. avium, M. intracellulare, and M. scrofulaceum. Phagocytosis was rapid and reached a maximum in 30 min. M. avium, M. intracellulare, and M. scrofulaceum grew within T. pyriformis, increasing by factors of 4- to 40-fold after 5 days at 30°C. Intracellular M. avium numbers remained constant over a 25-day period of growth (by transfer) of T. pyriformis. Intracellular M. avium cells also survived protozoan encystment and germination. The growth and viability of T. pyriformis were not affected by mycobacterial infection. The results suggest that free-living phagocytic protozoa may be natural hosts and reservoirs for M. avium, M. intracellulare, and M. scrofulaceum.
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MycoBacterium scrofulaceum: a bacterial contaminant in plant tissue culture
Plant Science, 1991Co-Authors: Ruth Ann Taber, Joseph O Falkinham, Margie A. Thielen, Roberta H. SmithAbstract:Abstract The identification of a widespread contaminant of plant cell cultures is reported. MycoBacterium scrofulaceum Prissick and Masson appears as yellow colonies on the medium and plant tissues of many ornamentals being propagated in vitro. It is a Gram positive and Acid Fast Bacterium and requires a specifically formulated medium for growth in pure culture. Many laboratories have alluded to this contaminant but have been unable to identify it because of its culture requirements.
Maryam Mohammadi Tashakkori - One of the best experts on this subject based on the ideXlab platform.
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Production of MPT-64 recombinant protein from virulent strain of MycoBacterium bovis
Iranian journal of veterinary research, 2018Co-Authors: Maryam Mohammadi Tashakkori, Majid Tebianian, Mohammad Tabatabaei, Nader MosavariAbstract:Tuberculosis (TB) is a zoonotic infectious disease common to humans and animals which has been caused by a rod shaped, Acid Fast Bacterium, called MycoBacterium bovis. The rapid and sensitive detection is a great challenge for TB diagnosis. The virulent strains of MycoBacterium tuberculosis complex (MTBC) have 16 different regions of difference (RD) in their genome which encode some important antigens. The major protein of M. bovis 64 (MPT-64) is one of the main immune-stimulating antigens which are encode by RD-2 region. The aim of the present study was cloning, expression and purification of MPT-64 as a protein antigen of M. bovis in a prokaryotic system for the usage in the future diagnostic studies. In this experimental study, the mpt-64 gene with 687 bp has been proliferated from M. bovis whole genome by polymerase chain reaction (PCR) method. The PCR product has been digested by BamHI and HindIII restriction enzymes and cloned into pQE-30 plasmid. The recombinant protein has been expressed in the Escherichia coli M15 with induction by isopropyl-β-D-thiogalactopyranoside (IPTG). The expressed protein was analyzed on SDS-PAGE, and purified with Nitrilotriacetic Acid (Ni-NTA) column. Finally, its biological properties were confirmed in Western blotting method using specific antibodies. Data showed the successful cloning of mpt-64 gene (as a 687 bp segment) in expression vector. The MPT-64 recombinant protein was ideally expressed and purified as a 24 kDa protein. The result of this study indicated that MPT-64 recombinant protein (24 kDa) has been successfully expressed and purified in a prokaryotic system, so this protein could be used for differential diagnosis of pathogenic and non-pathogenic MycoBacterium, in suspected BTB cases.
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Cloning, expression, and purification of recombinant protein MPT-64 from a virulent strain of MycoBacterium bovis in a prokaryotic system.
International journal of mycobacteriology, 2016Co-Authors: Maryam Mohammadi Tashakkori, Majid Tebianian, Mohammad Tabatabaei, Nader MosavariAbstract:Tuberculosis (TB) is a zoonotic infectious disease common to humans and animals that is caused by the rod-shaped Acid-Fast Bacterium MycoBacterium bovis. Rapid and sensitive detection of TB is promoted by specific antigens. Virulent strains of the TB complex from M. bovis contain 16 regions of difference (RD) in their genome that encode important proteins, including major protein of MycoBacterium Tuberculosis 64 (MBT-64, which is a primary immune-stimulating antigen encoded by RD-2. In this study, we cloned, expressed, and purified MPT-64 as a potent M. bovis antigen in a prokaryotic system for use in future diagnostic studies. The antigenic region of the Mpt64 gene was investigated by bioinformatics methods, cloned into the PQE-30 plasmid, and expressed in Escherichia coli M15 cells, followed by isopropyl β-d-1-thiogalactopyranoside induction. The expressed protein was analyzed sodium dodecyl sulfate polyacrylamide gel electrophoresis and purified using a nickel-affinity column. Biological activity was confirmed by western blot using specific antibodies. Our data verified the successful cloning of the Mpt64 gene (687-bp segment) via the expression vector and purification of recombinant MPT-64 as a 24-kDa protein. These results indicated successful expression and purification of recombinant MPT-64 protein in a prokaryotic system. This protein can be used for serological diagnosis, improved detection of pathogenicity and non-pathogenicity between infected cattle, and for verification of suspected cases of bovine TB. Copyright © 2016.
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Cloning, expression, and purification of recombinant protein MPT-64 from a virulent strain of MycoBacterium bovis in a prokaryotic system.
International Journal of Mycobacteriology, 2016Co-Authors: Maryam Mohammadi Tashakkori, Majid Tebianian, Mohammad Tabatabaei, Nader MosavariAbstract:Abstract Objective Tuberculosis (TB) is a zoonotic infectious disease common to humans and animals that is caused by the rod-shaped Acid-Fast Bacterium MycoBacterium bovis. Rapid and sensitive detection of TB is promoted by specific antigens. Virulent strains of the TB complex from M. bovis contain 16 regions of difference (RD) in their genome that encode important proteins, including major protein of MycoBacterium Tuberculosis 64 (MBT-64, which is a primary immune-stimulating antigen encoded by RD-2. In this study, we cloned, expressed, and purified MPT-64 as a potent M. bovis antigen in a prokaryotic system for use in future diagnostic studies. Methods The antigenic region of the Mpt64 gene was investigated by bioinformatics methods, cloned into the PQE-30 plasmid, and expressed in Escherichia coli M15 cells, followed by isopropyl β- d -1-thiogalactopyranoside induction. The expressed protein was analyzed sodium dodecyl sulfate polyacrylamide gel electrophoresis and purified using a nickel-affinity column. Biological activity was confirmed by western blot using specific antibodies. Results Our data verified the successful cloning of the Mpt64 gene (687-bp segment) via the expression vector and purification of recombinant MPT-64 as a 24-kDa protein. Conclusion These results indicated successful expression and purification of recombinant MPT-64 protein in a prokaryotic system. This protein can be used for serological diagnosis, improved detection of pathogenicity and non-pathogenicity between infected cattle, and for verification of suspected cases of bovine TB.