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Radiotracer Labeling of Brain Slices
Published in Avital Schurr, Benjamin M. Rigor, BRAIN SLICES in BASIC and CLINICAL RESEARCH, 2020
George C. Newman, Frank E. Hospod, Clifford S. Patlak, Hui Qi
Wet weights are measured by placing the slice on a microscope slide wrapped with Teflon™ tape, removing all visible buffer with a triangular wedge of filter paper, covering the slice with a plastic cap from a 5-ml vial to prevent desiccation, and weighing on a balance with 0.01 mg resolution. After the slide is weighed, the slice is removed with a spatula to a labeled microfuge tube and promptly stored at −20°C. The slide with the cap is reweighed after carefully returning tissue residues adherent to the spatula back onto the Teflon™. The slice in the microfuge tube is then sonicated (Microson XL Ultrasonic Cell Disrupter with a 3/32 inch microprobe, Heat Systems, Farmingdale, NY) at 30% maximum power in 200 μl of 0.3 M perchloric acid with three 1 s pulses or until complete dispersion. The tip is rinsed in an additional 200 μl of perchloric acid. The combined fractions are centrifuged at 11,000 × g for 10 min to produce the protein pellet. Prior to obtaining the cell disrupter, slices are homogenized by hand in 3 aliquots totalling 200-μl using glass microtissue grinders (#885470–0000, Kontes, Vineland, NJ) maintained below −20°C in a sample box cooled to −80°C in a deep freezer. The two methods of tissue preparation produce equivalent results. The supernatant is analyzed if necessary or discarded, and the pellet is digested in 500 μl of 1 N NaOH at 60°C for 1 h. The digested sample is neutralized with 500 μl of 1 N HCl. For a 500-μm slice, 200-μl aliquots are analyzed for protein by the method of Lowry and coworkers.8
Technetium-Labeled Compounds
Published in Garimella V. S. Rayudu, Lelio G. Colombetti, Radiotracers for Medical Applications, 2019
Suresh C. Srivastava, Powell Richards
If the precontact of the solution with mercury has not been carefully avoided, the data in hydrochloric and sulfuric acid media often show equal intensity waves I (4e−) and II (3e−). The relative heights, however, are in better agreement in perchloric acid media. The half-wave potential for the first wave is not affected significantly by the presence of complexing agents, the rate-determining step being:
Experimental Protocols for Generation and Evaluation of Articular Cartilage
Published in Kyriacos A. Athanasiou, Eric M. Darling, Grayson D. DuRaine, Jerry C. Hu, A. Hari Reddi, Articular Cartilage, 2017
Kyriacos A. Athanasiou, Eric M. Darling, Grayson D. DuRaine, Jerry C. Hu, A. Hari Reddi
Note: Perchloric acid is very dangerous. Use only in a chemical wash-down hood with glass pipettes, and wear appropriate personal protective equipment at all times. Make sure tubes used to contain the solutions are autoclavable.
Development of paracetamol-caffeine co-crystals to improve compressional, formulation and in vivo performance
Published in Drug Development and Industrial Pharmacy, 2018
Sumera Latif, Nasir Abbas, Amjad Hussain, Muhammad Sohail Arshad, Nadeem Irfan Bukhari, Hafsa Afzal, Sualeha Riffat, Zeeshan Ahmad
For drug analysis, to 1 ml of plasma, 60 µl of 50% perchloric acid was added. This mixture was vortexed for 5 min and centrifuged at 13,500 rpm for 15 min. Supernatant was filtered using 0.22 µm syringe filter. A 50 µl of clear filtrate was injected into the HPLC with a run time of 10 min and drug content was analyzed with a previously reported method [39]. The HPLC system (Shimadzu 20A, Japan) was equipped with a LC-20AT VP pump, a SIL-20AC HT auto sampler, SPD-M20A detector (photodiode array), CTO 20 AC column oven, CBM 20A controller unit and a reversed phase symmetry C18 (4.6 × 250 mm, 5-µm particle-size) column. The mobile phase was composed of water, methanol, and acetonitrile (80:10:10, v:v:v), filtered through a 0.45 membrane filter and was delivered at a flow rate of 1 ml/min. The analysis was carried out under isocratic conditions maintaining column temperature at 40 °C. A photodiode array detector set at 245 nm was used for recording chromatograms. Calibration curve (R2 = 0.9974) prepared for PCM was based on peak area measurements of standard solutions and spiked plasma samples for concentrations of 0.1, 0.5, 1.0, 5.0, 10.0, and 15.0 µg/ml. The limits of detection and quantification were 0.03 and 0.1 µg/ml, respectively.
Association between TPMT*3C and decreased thiopurine S-methyltransferase activity in patients with neuromyelitis optica spectrum disorders in China
Published in International Journal of Neuroscience, 2018
Xiaoqing Gong, Shenghui Mei, Xindi Li, Xingang Li, Heng Zhou, Yonghong Liu, Anna Zhou, Li Yang, Zhigang Zhao, Xinghu Zhang
6-mercaptopurine monohydrate (lot:3B3PG-PN, >98% purity) was purchased from Tokyo Chemical Industry Co. Ltd. (Japan). 6-MMP (lot:20130904, 97% purity) was obtained from Sinopharm Chemical Reagent Co. Ltd. (China). Methotrexate (internal standard (IS), lot: 100138-201104, 99.4% purity) was purchased from the National Institutes for Food and Drug Control (China). S-adenosyl-L-methionine (lot: L240P07, 98% purity) was purchased from Beijing J & K Chemical Technology Co. Ltd. (China). Allopurinol (lot: 081M1112V) was obtained from Sigma-Aldrich Co. (USA). Perchloric acid (lot: E131216H, 70%–72% purity) was purchased from Tianjin Xinyuan Chemical Co. Ltd. (China). Methanol and formic acid were obtained from Fisher Scientific (USA). Ultrapure water was generated from Milli-Q water purification device (Millipore, USA). Drug-free haemolysates were obtained from healthy volunteers.
Anti-hepatofibrosis effect of Allium senescens in activated hepatic stellate cells and thioacetamide-induced fibrosis rat model
Published in Pharmaceutical Biology, 2018
Gwang-Mo Shin, Sushruta Koppula, Yun-Jin Chae, Hyun-Su Kim, Jae-Dong Lee, Myong-Ki Kim, MinDong Song
TAA, silymarin, hydroxyproline, p-dimethylaminobenzaldehyde, 1,1,3,3-tetraethoexypropane, chloramine-T, 5,5-dithiobis-2-nitrobenzoic acid (DTNB), and glutathione (GSH), standards of chlorogenic acid, p-coumaric acid, and rutin were purchased from Sigma (St. Louis, MO). Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were acquired from Invitrogen (Carlsbad, CA). Perchloric acid was obtained from GFS Chemical Co. (Columbus, OH). GOT-GPT assay kit was purchased from Asan Pharmaceutical (Hwaseong-si, Korea). Annexin V-FITC and propidium iodide (PI) Apoptosis Detection Kit I was acquired from BD Biosciences (San Jose, CA). All other reagents used in this study were of the highest grade available commercially.