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Drug-loaded nanofibers (200 mg) were placed in 900 mL of physiological saline (PS; 0.9 wt%) at 37°C ± 1°C.
Wistar male rats, age of which was 24 months at the beginning of the experiments, were daily administered intraperitoneally with FeSNO, 5 mg/kg of mass in 1.0 ml of physiological solution (experimental animals) or administered with 1.0 ml of physiological solution (control animals) for 14 days.
Furthermore, the adult rats of age of 7-8 months were subjected to the acute irradiation with X-rays (5 Gy dose) after which the irradiated animals were intraperitoneally administered daily for 30 days with FeSNO, 5 mg/kg of mass in 1.0 ml of physiological solution (experimental animals) or with 1.0 ml of physiological solution (control animals).
After the 3 h adhesion phase and the washing step, the gold surfaces were transferred into a sterile tube containing 2 mL of physiological sterile solution (0.9% NaCl), then sonicated 3 min at 60 W.
Rats received multiple intraperitoneal injections of 1 mL of physiological saline (0.9 % NaCl) for the control (placebo) group and 1 mL of nanoparticle suspensions in physiological saline for the treatment groups.
After rinsing with 3 mL of water, [18F]FFMZ was eluted with 1 mL of 70% EtOH, diluted with 10 mL of physiological saline, and sterilized by filtration (0.22 μm Millex-FG).
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Both control groups (groups B and D) also received 100 mL infusion of physiological saline over a 15-minute period before surgery.
The PBMC layer was washed in 40 mL RPMI, resuspended in 10 mL RPMI and 10% fetal bovine serum (FBS), and centrifuged at 300 g 10 min at 22°C and the pellet was resuspended in 2 mL aliquots of physiological salt solution.
For each strain/biomass, 1 mL was removed sterilely from the culture flask and diluted in 9 mL of sterile physiological salt solution, and repeated to dilutions of 10−2, 10−3, 10−4, and 10−6.
Primary decimal dilutions were prepared by adding 225 ml of sterile physiological peptone saline (PPS, 0.85 g NaCl and 1 g neutralized bacteriological peptone (Oxoid, Basingstoke, U.K). per L) to each of the 25 g samples.
All biopsy and needle retrieval specimens were homogenized and mixed in 1.0 ml of sterile physiological saline, diluted serially in saline in triplicate, and then plated on tryptic soy agar for colony counts after incubation at 37°C.
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