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MEPH was orally administered to the test animals 30 min before the experiments at the doses of 25, 50, and 75 mg/kg body weight in both the chemical-induced and heat-induced pain models.
All animals were maintained in groups (5 rats per cage), previously housed to laboratory conditions during one week before the experiments at 25 ± 3°C on a 12-h light/dark cycle, and had access to food and water ad libitum during all the experimental days.
All animals were maintained in groups (5 rats per cage), previously housed to laboratory conditions during one week before the experiments at 25 ± 3°C on a 12 h light/dark cycle and had access to food and water ad libitum.
The animals arrived in the laboratory 1 week before the experiments (at 7 weeks of age) and were housed in groups of four of the same strain in standard plastic cages (Type IV Makrolon®), under controlled conditions of light: dark cycle (12 12 h, lights on at 07:00 h).
MECN was dissolved in DMSO and H2O in 2 1 ratio respectively and orally administered to the test animals 30 min before the experiments at the doses of 50, 100, and 200 mg/kg body weight in both the chemical- and heat-induced pain models.
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They were transferred and cultured anaerobically (10% H2, 5% CO2, 85% N2) on Schaedler agar plates (Oxoid, Basingstoke, GB) containing 8% sheep blood 24 h before starting the experiments at 37°C.
For electron microscopy HeLa and HEp-2 cells were seeded the day before the experiment at a density of 4×105 cells per 25 cm25
The fish were acclimated to the ponds for approximately 14 months before the experiment, at which time the fish were 15 months old.
Cells were incubated with H89 (Sigma), and Orlistat (Sigma) in regular media for 1 h before the experiment at the concentrations of 5 and 100 μM respectively.
Briefly, 2 × 10/well CLL MSCs were seeded and incubated for a few days before the experiment at 37 °C in 5% CO2 up to confluence.
Subjects were admitted to the research center the evening before the experiment, at which time the individuals with type 1 diabetes had two intravenous cannulas inserted with 1% lidocaine local anesthesia.
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