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Strikingly, dystrophic Sgca null mice lacking Nfix displayed a more oxidative phenotype at 3 weeks (Fig. 4d), consistent with the higher resistance to fatigue observed by treadmill test (Fig. 4a, b, Supplementary Movie 1).
Most importantly, mice lacking Nfix showed increased resistance to fatigue and better muscle performance with respect to controls, in keeping with our observation that these mice are characterized by a more oxidative phenotype.
To verify whether the improvements observed in dystrophic mice lacking Nfix could be also attributed to a more oxidative phenotype, we performed a succinate dehydrogenase (SDH) staining, which highlights in blue the presence of oxidative fibers.
This is achieved through a mechanism acting at different levels: Nfix is able not only to slow down the regenerative burst characterizing the disease18, but also to promote a shift to a more oxidative phenotype, that is known to confer protection from oxidative stress-induced damage13, 15.
Glycolytic skeletal muscles have a lower capillary density than oxidative muscles but can increase their capillary density and convert to a more oxidative phenotype when subject to chronic motor nerve stimulation (CMNS).
However, mutant mice progressively developed IIB myofiber-specific atrophy accompanied by a metabolic switch towards a more oxidative phenotype, muscular lipid accumulation, sarcomere disorganization and fibrosis.
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In addition, a shift towards more oxidative muscle phenotypes associated with the XX genotype could have a beneficial influence on health related phenotypes.
The results of this study show that niacin supplementation in sheep as a model for ruminants induces muscle fiber transition from type II (glycolytic) to type I (oxidative) being indicative of a change of the muscle's metabolic phenotype towards a more oxidative one.
However, in the TA of Sod1 D83G/D83G mice at 15 weeks of age, grouping of intensely stained type I slow oxidative muscle fibres was observed and by 52 weeks almost all of the muscle fibres in the TA of Sod1 D83G/D83G mice stain intensely for SDH, indicating an oxidative phenotype that is more characteristic of slow twitch, type I muscle fibres (Supplementary Material, Fig. S4A).
As skeletal muscle with an oxidative phenotype is believed to be more resistant to oxidative stress, a comparison of GAPDH levels within predominately oxidative and predominately glycolytic muscles following I/R would provide a means of testing the hypothesis that oxidative stress and protein modification plays a major role in the observed decrease in GAPDH pool levels.
With the onset of cell differentiation, exposure to 10 microM Ag+ evoked even greater inhibition of DNA synthesis and more oxidative stress, selectively impaired neurite formation without suppressing overall cell growth, and preferentially suppressed development into the acetylcholine phenotype in favor of the dopamine phenotype.
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