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While previous results already indicated a slight age-related increase in mutant frequency for the total brain [2], in this experiment such an increase was also statistically significant, i.e., from 5.7×10−5 at 7 months to 9.2×10−5 at 30 months of age (p = 0.0318).
The polymerase mutant frequency for each strand of the microsatellite (e.g., GT vs. CA) was determined separately from two independent polymerase reactions per strand.
Despite a small average increase in mutant frequency for smokers, an increased proportion of base substitutions at AT base pairs in smokers (p = 0.2) hinted at a smoking-associated shift in the mutation spectrum.
Mutant frequencies were nearly identical for all three groups at 30 days, while at 90 days the mutant frequency for the hepatocarcinogen 2,4-DAT (12.1 +/- 1.4 x 10 -5)) was significantly higher (p < 0.01) as compared to both age-matched (spontaneous) controls (5.7 +/- 2.9 x 10 -5)) and the 2,6-DAT-exposed group (5.7 +/- 2.4 x 10 -5)).
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The mean lacZ mutant frequency (MF) for the solvent control was approximately twofold greater than the spontaneous MF observed in liver tissue.
In the following years, other T-DNA activation tagging poplar populations were produced and screened for developmental abnormalities including alterations in leaf and stem structure as well as overall stature by Harrison et al. [ 21] The mutant frequency reported for the largest activation tagging poplar population (with 1,800 independent transgenic lines) was about 2.4%.
In the case of the glpT::MAR2xT strain, the mutant frequency was determined for tobramycin (4 µg/ml) and imipenem (4 µg/ml) to study the possible epistatic effects of resistance to fosfomycin and these two antibiotics.
Fourfold and twofold increases in mutant frequency were observed for 3-NBA and PhIP exposures, respectively, without the addition of any exogenous metabolic activation.
DNA was extracted and mutant frequencies determined for each of these subparts as well as for the remaining part of the brain at 7-, 19-and 30-months of age, and compared with that of the total brain at these age levels.
Following a 30-day fixation period, the experiment was terminated and lung DNA was extracted for mutant frequency and adduct determinations.
The MMS- or UV-induced mutant frequency is quite low for ΔdinB strains carrying any of the plasmid-borne DinB variants.
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