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closeReferee Comments: Referee 2
Posted by PLOS_ONE_Group on 17 Mar 2008 at 21:39 GMT
Referee 2's Review:
Despite improvements the manuscript encompasses data that are troubling in terms of their execution and interpretation. Specifically:
Figure 2:
Oxygen consumption measured after 3 hours increases in the muscle preparations, why? The data in 2G do not support a role for nitric oxide in oxygen consumption by mitochondria. There is no effect of L-NMMA under control or after insulin challenge. At apparently peak nitric oxide production (12 hours) oxygen consumption is decreases by approximately 60% than the controls. Addition of arginine increases the inhibition but it is unclear if this inhibition was statistically significantly different from the insulin-induced effect. Critically inclusion of L-NMMA did not increase oxygen consumption as expected. These observations directly contradict the findings in Figure 6.
Despite improvements, it remains a formidable task to review this manuscript because of the imprecise and at times awkward writing style. A good example is the rather long single sentence in the conclusion section of the abstract.
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N.B. These are the comments made by the referee when reviewing an earlier version of this paper. Prior to publication the manuscript has been revised in light of these comments and to address other editorial requirements.
RE: Referee Comments: Referee 2
jpoderoso replied to PLOS_ONE_Group on 19 Mar 2008 at 18:24 GMT
Opportunely, we answered Reviewer 2 requests. Significant differences in mitochondrial O2 utilization in the presence of NOS substrate L-arginine alone or plus LNMMA inhibitor (Fig 2G) and the calculation of mtNOS efficiency (page 2) clearly confirmed the insulin regulatory NO effects on O2 uptake. This effect was as well evidenced in the flow cytometry studies in Fig 3 and after silencing muscle nNOS in Figure 6.