China
Africa
Explore chapters and articles related to this topic
A-Z of Standardisation, Pre-Clinical, Clinical and Toxicological Data
Published in Saroya Amritpal Singh, Regulatory and Pharmacological Basis of Ayurvedic Formulations, 2017
Antioxidant: Superoxide radical scavenging activity was done by ethylene diamine tetra acetate and Nitro Blue Tetrazolium Chloride assays against ascorbic acid and R2 was 0.976 (EC50 = 77.5 ^g/ml). Ferrous reducing power was evaluated by Oyaizu method where R2 was 0.986. All studies showed that Amalakayas Rasayana possesses antioxidant activity (Samarakoon, Chandola and Shukla 2011b).
Serum Amyloid a (SAA) in Mammary Tissues with Inflammatory Processes and in Mammary Corpora Amylacea
Published in Gilles Grateau, Robert A. Kyle, Martha Skinner, Amyloid and Amyloidosis, 2004
M.J.M. Toussaint, C. Hogarth, T.K.A. Nguyen, E. Loeb, J. Balciute, V. Vivanco, A.M. van Ederen, S. Jacobsen, T.A. Niewold, E. Gruys
Amyloid fibrils were isolated from mammary corpora amylacea according to Pras et al 1968 (4). The corpora amylacea were purified after collection from frozen tissue samples corresponding to samples fixed for histology. This tissue was shown to be positive in Congo red stained sections. A part of the last pellet was kept in 6 M Guanidine/0.55 M TrisHCL pH 8.4 overnight and diluted 1:1 in sample buffer. This sample was put on a 15% SDS-urea gel. Thereafter the gel was blotted, using routine WB technique. The nitrocellulose membrane was stained with rabbit anti bovine AA (5) (1:1000) and goat anti rabbit alkaline phosphatase (DAKO, Denmark, 1:1000), followed by 4-nitro blue tetrazolium chloride and 5-bromo-4-chloro-3-indolylphosphate (NBT/BCIP, Roche, Germany).
Osteogenic and Angiogenic Synergy of Human Adipose Stem Cells and Human Umbilical Vein Endothelial Cells Cocultured in a Modified Perfusion Bioreactor
Published in Organogenesis, 2021
Fatemeh Mokhtari-Jafari, Ghasem Amoabediny, Mohammad Mehdi Dehghan, Sonia Abbasi Ravasjani, Massoumeh Jabbari Fakhr, Yasaman Zamani
ALP activity was measured to assess the osteoblastic phenotype of hASC seeded on BCP20/80 scaffolds. After 14 days of culture, the scaffolds were transferred to 24-well culture plates (Cellstar, Frickenhausen, Germany) and washed with PBS. The cells were lysed with cOmplete™ Lysis-M buffer to determine ALP activity and protein content. P-nitrophenyl-phosphate (Fluka, Poole, UK) at pH 10.3 was used as substrate for ALP. The absorbance was read at 405 nm using a ELISA reader (Stat Fax 2100, Awareness Technology Inc., USA). ALP activity was expressed as µmoles of p-nitrophenol formed per hour per milligram of cellular protein. After 14 days of culture, ALP activity was also visualized using nitro blue tetrazolium chloride/5-bromo-4-chloro-3-indolyl phosphate (NBT/BCIP; Roche, Mannheim, Germany) following the standard protocol. The amount of protein was determined by using a BCA Protein Assay reagent Kit (PierceTM, Rockford, III, USA), and the absorbance was read at 540 nm with a ELISA reader (Stat Fax 2100, Awareness Technology Inc., USA).
Expression of estrogen receptor alpha in response to stress and estrogen antagonist tamoxifen in the shell gland of Gallus gallus domesticus: involvement of anti-oxidant system and estrogen
Published in Stress, 2021
Mukesh Kumar Niranjan, Raj Kumar Koiri, Rashmi Srivastava
Superoxide dismutase (SOD) assay: The activity of superoxide dismutase (SOD) was determined as photochemical inhibition of nitro-blue tetrazolium chloride (NBT). A reaction mixture containing 2.25 mM methionine, 27 ml of 100 mM of phosphate buffer (pH 7.8), 1 ml of 2.25 mM NBT, 1.5 ml of 200 mM methionine, 1 M sodium carbonate (Na2CO3) and 3 mM EDTA was prepared. About 1.35 ml of reaction mixture was taken and 50 µl of tissue sample was added to it. The reaction starts after adding 200 µl of 60 µM riboflavin. The reaction mixture was illuminated in the presence of both light and dark then absorbance was measured at 560 nm. Enzyme activity was expressed in unit/mg protein. One unit of enzyme is defined as the amount of enzyme inhibiting the rate reaction by 50% (Beauchamp & Fridovich, 1971).
Generation of GM-CSF-producing antigen-presenting cells that induce a cytotoxic T cell-mediated antitumor response
Published in OncoImmunology, 2020
Hiroaki Mashima, Rong Zhang, Tsuyoshi Kobayashi, Yuichiro Hagiya, Hirotake Tsukamoto, Tianyi Liu, Tatsuaki Iwama, Masateru Yamamoto, Chiahsuan Lin, Ryusuke Nakatsuka, Yuta Mishima, Noriko Watanabe, Takashi Yamada, Satoru Senju, Shin Kaneko, Alimjan Idiris, Tetsuya Nakatsura, Hideki Ohdan, Yasushi Uemura
The cDNA fragment of murine Cd274 at cDNA positions 59–735 (GenBank accession number: NM_021893.3) was used for generation of sense or anti-sense RNA probes. Digoxigenin (DIG)-labeled RNA probes were prepared using DIG RNA Labeling Mix (Roche Diagnostics). Tissues were fixed with G-Fix (Genostaff), embedded in paraffin on CT-Pro20 (Genostaff) using G-Nox (Genostaff), as a less toxic solvent than xylene, and sectioned at 5 μm. In situ hybridization (ISH) was performed using the ISH Reagent kit (Genostaff) according to the manufacturer’s instructions. Paraffin-embedded sections were hybridized with the DIG-labeled RNA probes at 60°C for 16 h. After hybridization, sections were incubated with anti-DIG alkaline phosphate conjugate (Roche Diagnostics). The bound label was detected using alkaline phosphate color substrates nitro-blue tetrazolium chloride and 5-bromo-4-chloro-3ʹ-indolylphosphatase p-toluidine salt (Sigma-Aldrich). Sections were then counterstained with Kernechtrot Stain Solution (Muto Pure Chemicals, Tokyo, Japan) and mounted with G-Mount (Genostaff).