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Plants from Brazil Used Against Snake Bites
Published in Mahendra Rai, Shandesh Bhattarai, Chistiane M. Feitosa, Wild Plants, 2020
Jocimar de Souza, Bruna Stramandinoli Deamatis, Fernanda Mayumi Ishii, Ingrid Francine Araújo de Oliveira, Gustavo Rodrigues Toledo Piza, Jorge Amaral Filho, Edson Hideaki Yoshida, José Carlos Cogo, Angela Faustino Jozala, Denise Grotto, Rauldenis Almeida Fonseca Santos, Yoko Oshima-Franco
The diaphragm and its phrenic nerve branch were obtained from mice anesthetized with Halothane (Cristália®) and sacrificed by exsanguination. Hemidiaphragms were mounted under a tension of 0.5 g in a 5 mL organ bath (Bülbring 1997) containing Tyrode solution, and aerated with 95% O2 and 5% CO2. Tyrode solution maintains the physiological conditions of the neuromuscular preparation at pH 7.0, and consists of (in mM): 137 NaCl, 2.7 KCl, 1.8 CaCl2, 0.49 MgCl2, 0.42 NaH2PO4, 11.9 NaHCO3, and 11.1 Glucose. The preparation is indirectly stimulated through the phrenic nerve (ESF-15D double physiological stimulator), using supraximal stimuli and a frequency of 0.06 Hz with duration of 0.2 ms. Recording of muscle contraction is produced through the isometric cat. transducer 7003, coupled with a 2-Channel Recorder Gemini cat.7070, containing Basic Preamplifiers cat.7080 (Ugo Basile®). After recording under control conditions for 10 minutes during the stabilization of the preparation, the pharmacological protocols were performed.
Isolated Papillary Muscle Preparation
Published in John H. McNeill, Measurement of Cardiac Function, 2020
Preparation of normal and High Potassium Tyrode SolutionsPrepare the following salt solutions in the molarities indicated in parentheses. Stock solutions: Prepare 2 liters of concentrated (8x) Tyrode solution stock with the following composition: Combine these volumes in a 2-1 volumetric flask and bring up to 2000 ml with triple-distilled, deionized water.To make up 1 liter of regular Tyrode solution (calcium concentration = 2.4 mM): To make up 1 liter of high potassium Tyrode solution:
Host-Parasite Interactions With Macrophages In Culture
Published in Hans H. Gadebusch, Phagocytes and Cellular Immunity, 2020
Lee S. F. Soderberg, Morris Solotorovsky
In 1933, Baker8 examined the nutrition of macrophages in culture. Baker used explants from fragments of chick embryo spleen or blood leukocyte film as the source of macrophages and emphasized the importance of serum as a source of nutrients. The explants were fastened in Carrel flasks by means of plasma coagulum allowing the cells to form a colony. Cells from this colony migrated outwards and several days after migration the original fragment was excised. Following migration of cells onto the glass, the clot was excised. Tyrode’s solution with 25% homologous serum was used for maintenance of such preparations. The pH was adjusted with lactic acid, HC1, or with maintenance in a 5% C02 atmosphere. Monocytes proliferated most rapidly at pH 7.2 to 7.4. Increasing the concentration of the serum accelerated proliferation, but serum concentrations above 50% were toxic. Glutathione, nucleic acid, hemoglobin, sodium lactate, egg yolk, and albumin were not nutritive. Serum alone was entirely adequate for nutrition of the monocytes. Colonies of monocytes were cultivated for 2 months and proliferation was profuse, covering the entire surface of the flask in less than a month. Subcultures also continued to grow.
In vitro evaluation of intestinal absorption of tiliroside from Edgeworthia gardneri (Wall.) Meisn.
Published in Xenobiotica, 2021
Xiongwei Yin, Min Wang, Zhining Xia
In the absorption experiment of WAE with the Ussing chamber model, bilateral chambers were filled with Tyrode’s solution (5 mL). After balancing incubation for 15 min, the Tyrode’s solution was removed and replaced with the fresh Tyrode’s solution containing WAE (2 mg/mL) in the mucosal chamber. Fresh Tyrode’s solution (5 mL) was added to the other chamber. In the experiment of WAE and different inhibitors, 5 mL of Tyrode’s solution containing WAE (2 mg/mL) and inhibitors (1 mM) were filled in one mucosal side and 5 mL of Tyrode’s solution was added to the other chamber simultaneously. The inhibitors included verapamil (1 mM), rifampin (1 mM), and novobiocin (1 mM). Samples were collected from the serosal chamber every 15 min within 90 min and 500 μL of fresh Tyrode’s solution was supplemented after each sampling.
MRGPRX2 is critical for clozapine induced pseudo-allergic reactions
Published in Immunopharmacology and Immunotoxicology, 2021
Di Wei, Tian Hu, Ya-jing Hou, Xiang-jun Wang, Jia-yu Lu, Shuai Ge, Cheng Wang, Huai-zhen He
LAD2 cells were seeded into 96-well plate at 2 × 105 cells/well and incubated overnight. After that, the culture medium was removed. The drug prepared by Tyrode’s solution was added to each well and the cells were incubated for 30 min. Then, 96-well plate was centrifuged at 2000 g for 5 min to get 50 μL supernatant. To obtain the total β-hexosaminidase content, the cells in each well were lysed with 0.1% Triton X-100 and centrifuged again to obtain supernatant. Finally, the supernatant reacted with pnitrophenyl N-acetyl-β-D-glucosamide in 0.1 M citric acid/sodium citrate (pH = 4.5) for 90 min at 37 °C. The reaction was halted by stop buffer (0.1 M sodium carbonate/sodium bicarbonate, pH 11.0), and the sample was measured at 405 nm using microplate spectrophotometer.
Distinct effects of resveratrol on seizures and hyperexcitability induced by NMDA and 4-aminopyridine
Published in Nutritional Neuroscience, 2019
Ya-Jean Wang, Chung-Pin Hsieh, Ming-Huan Chan, Tzu-Yi Chan, Linyi Chen, Hwei-Hisen Chen
Evoked action potential firing was performed by holding membrane potential at −65 mV and then depolarizing cell through a positive current injection (ranging from 5 to 30 pA). To record the action potentials, the chamber solutions were applied with normal Tyrode’s solution. The composition of normal Tyrode’s solution was as follows (in mM): NaCl 136.5, KCl 5.4, CaCl2 1.8, MgCl2 0.53, glucose 5.5, and HEPES 5.5 (pH 7.4). The patch pipettes solutions contained (mM): KCl 145, MgCl2 1, Na2ATP 3, EGTA 0.1, and HEPES 5.5 (pH 7.2). The frequency of action potentials was measured in 6 seconds current clamp. Then, the NMDA receptor-dependent action potentials were evoked by co-application of NMDA (100 μM) and glycine (100 μM). 4-AP (100 μM) was added into the chamber to trigger the 4-AP-mediated action potentials.