The Experts below are selected from a list of 252 Experts worldwide ranked by ideXlab platform
Yuantsong Chen - One of the best experts on this subject based on the ideXlab platform.
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high level production of recombinant human lysosomal acid alpha glucosidase in chinese hamster ovary cells which targets to heart muscle and corrects glycogen accumulation in fibroblasts from patients with pompe disease
Proceedings of the National Academy of Sciences of the United States of America, 1996Co-Authors: Johan L K Van Hove, H W Yang, Jeryuarn Wu, R O Brady, Yuantsong ChenAbstract:Abstract Infantile Pompe disease is a fatal genetic muscle disorder caused by a deficiency of acid Alpha-Glucosidase, a glycogen-degrading lysosomal enzyme. We constructed a plasmid containing a 5'-shortened human acid Alpha-Glucosidase cDNA driven by the cytomegalovirus promoter, as well as the aminoglycoside phosphotransferase and dihydrofolate reductase genes. Following transfection in dihydrofolate reductase-deficient Chinese hamster ovary cells, selection with Geneticin, and amplification with methotrexate, a cell line producing high levels of the Alpha-Glucosidase was established. In 48 hr, the cells cultured in Iscove's medium with 5 mM butyrate secreted 110-kDa precursor enzyme that accumulated to 91 micrograms.ml-1 in the medium (activity, > 22.6 mumol.hr-1.ml-1). This enzyme has a pH optimum similar to that of the mature form, but a lower Vmax and Km for 4-methylumbelliferyl-alpha-D-glucoside. It is efficiently taken up by fibroblasts from Pompe patients, restoring normal levels of acid Alpha-Glucosidase and glycogen. The uptake is blocked by mannose 6-phosphate. Following intravenous injection, high enzyme levels are seen in heart and liver. An efficient production system now exists for recombinant human acid Alpha-Glucosidase targeted to heart and capable of correcting fibroblasts from patients with Pompe disease.
Johan L K Van Hove - One of the best experts on this subject based on the ideXlab platform.
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high level production of recombinant human lysosomal acid alpha glucosidase in chinese hamster ovary cells which targets to heart muscle and corrects glycogen accumulation in fibroblasts from patients with pompe disease
Proceedings of the National Academy of Sciences of the United States of America, 1996Co-Authors: Johan L K Van Hove, H W Yang, Jeryuarn Wu, R O Brady, Yuantsong ChenAbstract:Abstract Infantile Pompe disease is a fatal genetic muscle disorder caused by a deficiency of acid Alpha-Glucosidase, a glycogen-degrading lysosomal enzyme. We constructed a plasmid containing a 5'-shortened human acid Alpha-Glucosidase cDNA driven by the cytomegalovirus promoter, as well as the aminoglycoside phosphotransferase and dihydrofolate reductase genes. Following transfection in dihydrofolate reductase-deficient Chinese hamster ovary cells, selection with Geneticin, and amplification with methotrexate, a cell line producing high levels of the Alpha-Glucosidase was established. In 48 hr, the cells cultured in Iscove's medium with 5 mM butyrate secreted 110-kDa precursor enzyme that accumulated to 91 micrograms.ml-1 in the medium (activity, > 22.6 mumol.hr-1.ml-1). This enzyme has a pH optimum similar to that of the mature form, but a lower Vmax and Km for 4-methylumbelliferyl-alpha-D-glucoside. It is efficiently taken up by fibroblasts from Pompe patients, restoring normal levels of acid Alpha-Glucosidase and glycogen. The uptake is blocked by mannose 6-phosphate. Following intravenous injection, high enzyme levels are seen in heart and liver. An efficient production system now exists for recombinant human acid Alpha-Glucosidase targeted to heart and capable of correcting fibroblasts from patients with Pompe disease.
Dominique Sohounhloue - One of the best experts on this subject based on the ideXlab platform.
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In vitro biological effects of two anti-diabetic medicinal plants used in Benin as folk medicine.
BMC Complementary and Alternative Medicine, 2013Co-Authors: Fifa Bothon, Eric Debiton, Felicien Avlessi, Christiane Forestier, Jean-claude Teulade, Dominique SohounhloueAbstract:BACKGROUND: Extracts from Polygonum senegalensis (Polygonaceae) and Pseudocedrela kotschyi (Meliaceae) are two important traditionally used medicinal plants in rural Benin to treat many diseases and notably type 2 diabetes. The aim of the study was to investigate the Alpha-Glucosidase inhibition, antioxidant and antibacterial activities of those plants extract: Polygonum senegalensis leaves, and Pseudocedrela kotschyi root. METHODS: Hydro-alcoholic (50%) extracts were analyzed for their phytochemical content and tested for their inhibition potency on Alpha-Glucosidase from Saccharomyces cerevisiae. Antioxidant activities were assessed using the DPPH, ORAC, FRAP and DCFH-DA (cell based) assay. Finally, the antibacterial activity was evaluated using MIC determination on four Gram-positive cocci (Bacillus subtilis, Clostridium difficile, Enterococcus faecalis, Staphylococcus aureus), three Gram-negative bacilli (Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae), and the yeast Candida albicans. RESULTS: Each extract presented significant Alpha-Glucosidase inhibition and antioxidant activities. Polygonum senegalensis leaf extracts were the most active in each in vitro assay with an IC50 = 1.5 mug/ml for Alpha-Glucosidase inhibition and an IC50 = 6.8 mug/ml for DPPH scavenging, - 4.5 mumol Fe II/g of dry matter - 9366 mumol Trolox / g DW - for FRAP and ORAC values, respectively. IC50 = 2.3 mug GA / ml for DCFH-DA assay. Concerning its antibacterial activity, a growth inhibitory effect was observed only against three Gram negative bacilli: B. subtilis, E. faecalis, S. aureus and the yeast C. albicans at high concentration. CONCLUSION: The results showed that the semi alcoholic extract of the two studied plants possess Alpha-Glucosidase inhibitory activity, antioxidant potency, and low antibacterial effect.
Naoki Asano - One of the best experts on this subject based on the ideXlab platform.
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alkaloids from the poisonous plant ipomoea carnea effects on intracellular lysosomal glycosidase activities in human lymphoblast cultures
Journal of Agricultural and Food Chemistry, 2003Co-Authors: Kyoko Ikeda, Atsushi Kato, Isao Adachi, Mitsue Haraguchi, Naoki AsanoAbstract:: There is natural intoxication of livestock by the ingestion of Ipomoea carnea (Convolvulaceae) in Brazil and other parts of the world. The alkaloidal glycosidase inhibitors swainsonine, 2-epi-lentiginosine, and calystegines B(1), B(2), B(3), and C(1) have been identified as constituents of this plant. Swainsonine is a potent inhibitor of rat lysosomal alpha-mannosidase, with an IC(50) value of 0.02 microM, whereas calystegines B(1), B(2), and C(1) are potent inhibitors of rat lysosomal beta-glucosidase, with IC(50) values of 2.1, 0.75, and 0.84 microM, respectively. The action of swainsonine results in a lysosomal storage disorder that closely mimics alpha-mannosidosis in humans. To determine whether the toxicity of I. carnea to livestock is due to purely swainsonine or due to a combination of effects by swainsonine and calystegines, intracellular lysosomal glycosidase activities in normal human lymphoblasts grown with inhibitors in the medium were examined. Incubation of lymphoblasts with 0.1 microM swainsonine for 3 days resulted in approximately 60% reduction of alpha-mannosidase activity. On the other hand, calystegines B(2) and C(1) showed no inhibition of beta-glucosidase up to 1 mM; instead inclusion of calystegines B(2) and C(1) at 100 microM in the culture medium increased its activity by 1.5- and 1.6-fold, respectively. Calystegines B(2) and C(1) seem to act as chemical chaperones, enhancing correct folding of the enzyme and enabling smooth trafficking to the lysosome. The lysosomal beta-glucosidase inhibitory calystegines seem to have little risk of inducing intoxication of livestock.
R O Brady - One of the best experts on this subject based on the ideXlab platform.
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high level production of recombinant human lysosomal acid alpha glucosidase in chinese hamster ovary cells which targets to heart muscle and corrects glycogen accumulation in fibroblasts from patients with pompe disease
Proceedings of the National Academy of Sciences of the United States of America, 1996Co-Authors: Johan L K Van Hove, H W Yang, Jeryuarn Wu, R O Brady, Yuantsong ChenAbstract:Abstract Infantile Pompe disease is a fatal genetic muscle disorder caused by a deficiency of acid Alpha-Glucosidase, a glycogen-degrading lysosomal enzyme. We constructed a plasmid containing a 5'-shortened human acid Alpha-Glucosidase cDNA driven by the cytomegalovirus promoter, as well as the aminoglycoside phosphotransferase and dihydrofolate reductase genes. Following transfection in dihydrofolate reductase-deficient Chinese hamster ovary cells, selection with Geneticin, and amplification with methotrexate, a cell line producing high levels of the Alpha-Glucosidase was established. In 48 hr, the cells cultured in Iscove's medium with 5 mM butyrate secreted 110-kDa precursor enzyme that accumulated to 91 micrograms.ml-1 in the medium (activity, > 22.6 mumol.hr-1.ml-1). This enzyme has a pH optimum similar to that of the mature form, but a lower Vmax and Km for 4-methylumbelliferyl-alpha-D-glucoside. It is efficiently taken up by fibroblasts from Pompe patients, restoring normal levels of acid Alpha-Glucosidase and glycogen. The uptake is blocked by mannose 6-phosphate. Following intravenous injection, high enzyme levels are seen in heart and liver. An efficient production system now exists for recombinant human acid Alpha-Glucosidase targeted to heart and capable of correcting fibroblasts from patients with Pompe disease.