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The thiol concentration for the incubation medium with bacteria only was the lowest, and this served as the baseline value.
Scanning the electrode between −0.7 to 0.3 V (vs. Ag/AgCl) at 70 mV/s for 50 cycles (∼24 min), placed in the growth medium with bacteria, significantly increased microbial adhesion when compared to SPCE without electrochemical stimulation.
The effect of PK 11195 on the cytotoxicity of P. fluorescens MF37 was determined through measurement of nitrite (NO2−) and lactate dehydrogenase (LDH) release by glial cells following a 24 h incubation in culture medium with bacteria.
Liquid medium with bacteria was gently removed from the wells, which were washed twice with phosphate buffered saline (PBS) 1×, pH 7.2 (Invitrogen, Grand Island, New York) to eliminate unbound bacteria without disturbing the adherent biofilm.
Medium with bacteria and 1% DMSO only were used as controls.
In addition, plates were centrifuged for 30 s at 1000 rpm to collect larvae (L1/L2) and the medium with bacteria at the bottom of the block plates.
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As stated elsewhere, since the cells were cultured in a sealed bag using a serum-free medium, contamination with bacteria or viruses was prevented, but all the cells underwent security tests before transfer to the patients.
Upon completion of the incubation, the medium with planktonic bacteria was removed and 10nmolL−1 of Molsidomine or MAHMA nonoate were added to the wells with biofilms.
To seed the porous medium with landfill bacteria, a mixture of Keele Valley Landfill and synthetic leachate permeated through the column under anaerobic conditions for the first 9 days of operation.
Upon completion of the incubation, the medium with planktonic bacteria was removed and serial dilutions of nitric oxide donors in PBS (in 200 μL) were added to the wells with biofilms.
Upon completion of the incubation, the medium with planktonic bacteria was removed and serial dilutions of nitric oxide donors in PBS or CNC (in 200 µL) were added to the wells with biofilms.
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