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David Read - One of the best experts on this subject based on the ideXlab platform.
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The use of the fluorescent dye, 3 ,3'-dihexyloxacarbocyanine iodide, for selective staining of ascomycete fungi associated with liverwort rhizoids and ericoid mycorrhizal roots
New Phytologist, 1991Co-Authors: Jeffrey G. Duckett, David ReadAbstract:SUMMARY The lipophilic fluorochrome 3,3'dihexyloxacarbocyanine iodine [DiOC6(3)], previously used to visualize mitochondria and ER in animal and plant cells, when applied at concentrations of 0-01-5 ,tg ml-l selectively stains ascomycetous hyphae in ericaceous roots and in the rhizoids of liverworts in the families Lepidoziaceae (both tropical and temperate species), Calypogeiaceae, Adelanthaceae, Cephaloziaceae and Cephaloziellaceae. Basidiomycetes forming endophytic associations with liverworts and ectomycorrhizas in seed plants, are stained with DiOC6(3) only at concentrations at and above 50 ,tg ml-'. VA mycorrhizal fungi in liverworts, pteridophytes and angiosperms fail to stain. Hyphae of the ericoid mycorrhizal fungus, Hymenoscyphus ericae, grown in axenic culture, are stained with much lower concentrations of DiOC6(3) than are those of a range of ectomycorrhizal fungi, an orchid fungus and Oidiodendron griseum which has been reported occasionally to form ericoid mycorrhizal associations. In contrast to other fluorescent probes that recognize fungal wall components, DiOC6(3) is a vital stain of fungal cytoplasm. Greater membrane permeability, compared to that in other fungi, is the likely basis for the selective staining of Hymenoscyphus ericae and the root and rhizoid-inhabiting ascomycetes with this dye. DiOC6(3) offers a rapid means for identifying intracellular ascomycetous mycorrhizas and for determining the distribution of living hyphae within these associations.
Susanti Susanti - One of the best experts on this subject based on the ideXlab platform.
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Aktivitas Antihiperurisemia Ekstrak Tanaman Jelatang (Urtica dioca L.) pada Mencit
'Poltekkes Kemenkes Kendari', 2021Co-Authors: Fadilah, Nitya Nurul, Susanti SusantiAbstract:Hyperuricemia is a high level of uric acid in the blood and can cause gout. One of the drugs used to treat hyperuricemia is allopurinol. The use of these drugs can cause various side effects if used for a long period of time. Nettle plants are known to contain flavonoids which can reduce uric acid levels in the blood. This study was conducted to test the activity of nettle plants and to find out the effective dose in reducing blood uric acid levels in mice. In this study, extracts were made using maceration method using 70% ethanol. Tests were carried out on mice induced by pottasium oxonate and chicken liver juice to increase uric acid levels. Then the test animals were divided into 5 groups cosisted of the negative control group (0.5% NaCMC), positive control (allopurinol 13mg/kgBW), the dose I group (125mg/kgBW), dose II (250mg/kgBW), and dose III (500mg/kgBW). Next step is to measured the decrease of uric acid levels in mice’s blood every hour for 5 hours. Then the data of decreased in uric acid levels were processed statistically using the Anova and Duncan method. Based on the results of test in mice, nettle extract has antihyperuricemia activity with successive doses of 500 mg/kgBW (89.91%), 250 mg/kgBW (73.7%), and 125mg/kgBW (63.56%).Hiperurisemia merupakan tingginya kadar asam urat dalam darah dan dapat menimbulkan penyakit gout. Salah satu obat yang digunakan untuk mengatasi hiperurisemia adalah allopurinol. Penggunaan obat tersebut menimbulkan berbagai efek samping jika digunakan dalam jangka waktu yang lama. Tumbuhan jelatang diketahui mengandung flavonoid yang dapat menurunkan kadar asam urat dalam darah. Studi ini dilakukan untuk menguji aktivitas tanaman jelatang dan untuk mengetahui dosis efektifnya dalam menurunkan kadar asam urat dalam darah pada mencit. Pada penelitian ini, ekstrak dibuat dengan menggunakan metode maserasi menggunakan etanol 70%. Uji dilakukan pada mencit yang diinduksi kalium oksonat dan jus hati ayam agar menaikkan kadar asam uratnya. Kemudian hewan uji dibagi menjadi 5 kelompok yakni terdiri dari kelompok kontrol negatif (NaCMC 0,5%BB), kontrol positif (allopurinol 13mg/kgBB), kelompok dosis I (125mg/kgBB), dosis II (250mg/kgBB), dan dosis III (500mg/kgBB). Selanjutnya diukur penurunan kadar asam urat darah mencit pada tiap jam selama 5 jam. Kemudian data penurunan kadar asam urat diolah secara statistik menggunakan metode Anova dan Duncan. Berdasarkan hasil uji pada mencit, ekstrak jelatang memiliki aktivitas antihiperurisemia dengan dosis berturut-turut adalah 500mg/kg BB (89,91%), 250mg/kg BB (73,7%), dan 125 mg/kg BB (63,56%)
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Aktivitas Antihiperurisemia Ekstrak Tanaman Jelatang (Urtica dioca L.) pada Mencit
'Poltekkes Kemenkes Kendari', 2020Co-Authors: Fadilah, Nitya Nurul, Susanti SusantiAbstract:Hyperuricemia is a high level of uric acid in the blood and can cause gout. One of the drugs used to treat hyperuricemia is allopurinol. The use of these drugs can cause various side effects if used for a long period of time. Nettle plants are known to contain flavonoids which can reduce uric acid levels in the blood. This study was conducted to test the activity of nettle plants and to find out the effective dose in reducing blood uric acid levels in mice. In this study, extracts were made using maceration method using 70% ethanol. Tests were carried out on mice induced by pottasium oxonate and chicken liver juice to increase uric acid levels. Then the test animals were divided into 5 groups cosisted of the negative control group (0.5% NaCMC), positive control (allopurinol 13mg/kgBW), the dose I group (125mg/kgBW), dose II (250mg/kgBW), and dose III (500mg/kgBW). Next step is to measured the decrease of uric acid levels in mice’s blood every hour for 5 hours. Then the data of decreased in uric acid levels were processed statistically using the Anova and Duncan method. Based on the results of test in mice, nettle extract has antihyperuricemia activity with successive doses of 500 mg/kgBW (89.91%), 250 mg/kgBW (73.7%), and 125mg/kgBW (63.56%). Catatan PenerbitPolekkes Kemenkes Kendari menyatakan tetap netral sehubungan dengan klaim dari perspektif atau buah pikiran yang diterbitkan dan dari afiliasi institusional manapun. PendanaanPenelitian ini didanai oleh Universitas Perjuangan Tasikmalaya.Hiperurisemia merupakan tingginya kadar asam urat dalam darah dan dapat menimbulkan penyakit gout. Salah satu obat yang digunakan untuk mengatasi hiperurisemia adalah allopurinol. Penggunaan obat tersebut menimbulkan berbagai efek samping jika digunakan dalam jangka waktu yang lama. Tumbuhan jelatang diketahui mengandung flavonoid yang dapat menurunkan kadar asam urat dalam darah. Studi ini dilakukan untuk menguji aktivitas tanaman jelatang dan untuk mengetahui dosis efektifnya dalam menurunkan kadar asam urat dalam darah pada mencit. Pada penelitian ini, ekstrak dibuat dengan menggunakan metode maserasi menggunakan etanol 70%. Uji dilakukan pada mencit yang diinduksi kalium oksonat dan jus hati ayam agar menaikkan kadar asam uratnya. Kemudian hewan uji dibagi menjadi 5 kelompok yakni terdiri dari kelompok kontrol negatif (NaCMC 0,5%BB), kontrol positif (allopurinol 13mg/kgBB), kelompok dosis I (125mg/kgBB), dosis II (250mg/kgBB), dan dosis III (500mg/kgBB). Selanjutnya diukur penurunan kadar asam urat darah mencit pada tiap jam selama 5 jam. Kemudian data penurunan kadar asam urat diolah secara statistik menggunakan metode Anova dan Duncan. Berdasarkan hasil uji pada mencit, ekstrak jelatang memiliki aktivitas antihiperurisemia dengan dosis berturut-turut adalah 500mg/kg BB (89,91%), 250mg/kg BB (73,7%), dan 125 mg/kg BB (63,56%). Catatan PenerbitPolekkes Kemenkes Kendari menyatakan tetap netral sehubungan dengan klaim dari perspektif atau buah pikiran yang diterbitkan dan dari afiliasi institusional manapun. PendanaanPenelitian ini didanai oleh Universitas Perjuangan Tasikmalaya
Mariehelene Ratinaud - One of the best experts on this subject based on the ideXlab platform.
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sensitive and reliable jc 1 and toto 3 double staining to assess mitochondrial transmembrane potential and plasma membrane integrity interest for cell death investigations
Cytometry Part A, 2003Co-Authors: Thomas Zuliani, Raphael E Duval, Chantal Jayat, Sylviane Schnebert, Patrice Andre, Marc Dumas, Mariehelene RatinaudAbstract:Background Apoptosis is currently studied by flow cytometry with mitochondrial membrane potential (Δψmt) and membrane integrity fluorochromes. Rhodamine 123 and DiOC6(3) remain controversial to identify cells displaying a low Δψmt. JC-1 constitutes a good Δψmt indicator, due to a fluorescence shift from green to orange emission, according to the increase in Δψmt. Nevertheless, it is not feasible to analyze it simultaneously with propidium iodide. Among available fluorescent probes, TOTO-3 seems to be a good candidate for double staining with JC-1. Methods Cell death of HaCaT cells was induced by H2O2 and FasL. Samples were stained with DiOC6(3)/IP or JC-1/TOTO-3 then analyzed by flow cytometry. Results were supported by confocal microscopy analyses of mitochondrial membrane potential. Moreover, cell morphology was determined on the sorted subpopulations defined on the basis of staining (JC-1 versus TOTO-3). Results We found that JC-1 is a more efficient mitochondrial probe than DiOC6(3). After stress induction, the fluorescence level of JC-1 and TOTO-3 clearly defined three fluorescent subpopulations, respectively: (1) JC-1high and TOTO-3low, (2) JC-1low and TOTO-3medium, and (3) JC-1low and TOTO-3high. Their morphologic aspects after cell sorting indicated that they corresponded to three functional states (intact, apoptotic, and necrotic cells), and data were supported by caspase activity measurements. Conclusions We propose a reliable and efficient staining, with JC-1 and TOTO-3 to discriminate three functional cellular states: intact, apoptotic, and necrotic/late apoptotic cells by flow cytometry. Cytometry Part A 54A:100–108, 2003. © 2003 Wiley-Liss, Inc.
Eberhard Spiess - One of the best experts on this subject based on the ideXlab platform.
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tomography of cells by confocal laser scanning microscopy and computer assisted three dimensional image reconstruction localization of cathepsin b in tumor cells penetrating collagen gels in vitro
Journal of Histochemistry and Cytochemistry, 1997Co-Authors: Anjarose Strohmaier, Torsten Porwol, H Acker, Eberhard SpiessAbstract:We used the nondestructive procedures of confocal laser scanning microscopy in combination with computer-assisted methods to visualize tumor cells in the process of penetrating collagen gels. Three independent sets of images were collected. The image information of all data sets was combined into one image, giving a three-dimensional (3D) impression at high light microscopic resolution and sensitivity. We collected information about the extracellular matrix using the reflection mode, the cell surface/morphology by staining with the fluorescent dye DiOC6(3), and the distribution of cathepsin B by Cy-3-labeled immunolocalization. The specific aim of our study was visualization of the spatial relationship of cell organelles as far as they contain the enzyme cathepsin B to cell morphology and motility in a 3D model of extracellular matrix. The majority of the enzyme was localized pericellularly, with no visible relationship to the direction of movement. However, substantial amounts also appeared in intramatri...
Jeffrey G. Duckett - One of the best experts on this subject based on the ideXlab platform.
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The use of the fluorescent dye, 3 ,3'-dihexyloxacarbocyanine iodide, for selective staining of ascomycete fungi associated with liverwort rhizoids and ericoid mycorrhizal roots
New Phytologist, 1991Co-Authors: Jeffrey G. Duckett, David ReadAbstract:SUMMARY The lipophilic fluorochrome 3,3'dihexyloxacarbocyanine iodine [DiOC6(3)], previously used to visualize mitochondria and ER in animal and plant cells, when applied at concentrations of 0-01-5 ,tg ml-l selectively stains ascomycetous hyphae in ericaceous roots and in the rhizoids of liverworts in the families Lepidoziaceae (both tropical and temperate species), Calypogeiaceae, Adelanthaceae, Cephaloziaceae and Cephaloziellaceae. Basidiomycetes forming endophytic associations with liverworts and ectomycorrhizas in seed plants, are stained with DiOC6(3) only at concentrations at and above 50 ,tg ml-'. VA mycorrhizal fungi in liverworts, pteridophytes and angiosperms fail to stain. Hyphae of the ericoid mycorrhizal fungus, Hymenoscyphus ericae, grown in axenic culture, are stained with much lower concentrations of DiOC6(3) than are those of a range of ectomycorrhizal fungi, an orchid fungus and Oidiodendron griseum which has been reported occasionally to form ericoid mycorrhizal associations. In contrast to other fluorescent probes that recognize fungal wall components, DiOC6(3) is a vital stain of fungal cytoplasm. Greater membrane permeability, compared to that in other fungi, is the likely basis for the selective staining of Hymenoscyphus ericae and the root and rhizoid-inhabiting ascomycetes with this dye. DiOC6(3) offers a rapid means for identifying intracellular ascomycetous mycorrhizas and for determining the distribution of living hyphae within these associations.