This study is worthwhile, and shows some interesting data to indicate the use of Opuntia for the prevention and treatment of acetaminophen-induced toxicity. Comments as follows: 1. The authors should explain how the juice was extracted from the fruit. Was all the juice extracted in one single procedure? If the extraction was done in batches, how did the authors ensure consistency between the batches? The authors should also include the yield from the fruit – ie. how much fruit was required to produce a dose of juice? 2. On what basis was the dose of 800mg/kg selected for the in vivo study? It would seem to be a very high dose. As the authors compare the effects with 50mg/kg GSH, it is important that they discuss the dose used. When comparing the effects of GSH and Opuntia, they need to include ‘at the doses used’. 3. P. 5, lines 164 – 165: For the in vitro study, can the authors clarify how the concentrations were calculated. If 125ul of extract was used, how did the authors translate that to 3.3 and 8.4mmol/L, and is this a valid and accurate translation? 4. P. 5, line 162: Why was FCS excluded from the medium? 5. While the in vitro study examines both prophylactic and therapeutic treatment, the in vivo study only investigates prophylactic treatment. How relevant is prophylactic treatment to the clinical situation of acetaminophen toxicity? The authors should include this in the discussion. 6. Table 1: As the flavonoids are a group of phenolic compounds, it would be expected that the Total Phenolic Content would be higher than the quantity of flavonoids. Can the authors explain why the quantity of flavonoids is higher than TPC for Opuntia robusta.
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This is a good preclinical study on a novel food supplement. It provides good evidence that sake protein may improve exercise endurance and that it may reduce fatigue. It provides a strong impetus for the study of this food supplement in humans. There are minor improvements that can be made on this manuscript, as follows: Introduction: The paragraphs need to be integrated better. Paragraph on fatigue seems out of place. Page 3, line 102: Sedentary controls received vehicle by gavage. Did PET group receive vehicle? Page 3, line 103: do the authors mean ‘the same volume of solution [relative] to individual body weight’? Page 3, line 111: the human equivalent dose [of] 18.5 g/60 kg (0.308 g/kg) = 0.308 = a mouse dose of x 12.3 = 3.8 g/kg Page 4, line 131: metal bar Page 4, line 141: For the forced swim test, can the authors specify how they defined exhaustion during the test? They mention possible drowning… can the authors clarify this please. Page 5, line 152: Please specify the method used for the sacrifice of the animals. Page 7, line 185: The body weights of the 3 groups are identical. Is this correct? Also, figure 1 is written in brackets but should this be figure 2? Page 7, lines 196 onwards: Where data is written in table 2, there is no need to give it again in the text. Also, the authors may consider adding another column in table 2 to give the p values from the ANOVAs. This would make it clearer. Page 7, lines 202 – 204. This sentence needs to be written more clearly. The average relative EFP weights of the PET and PET+SP groups were 31.0% and 33.5% lower than controls, respectively.
The methods do not mention that the EFPs were removed and measured. The authors should detail the methods used. Page 9, line 231: ‘demonstrates’ would be better than ‘reveals’ here. Page 10, line 248, 249: For clarity, the authors could add in – but [as was seen from the grip strength experiment], there was no effect on explosive force production. Page 10, line 249 - 251: The authors are firm in their assertion of the mechanism of SP. As the mechanism was not investigated in this study, I think the authors cannot be certain of this. They can however suggest that this may be the mechanism of action. Page 11, line 286: insert ‘statistically significant’ differences between groups. Page 12, line 294: Clarify/change the following sentence: ‘Urea is formed in the liver and is carried by the blood to the kidneys for excretion and an important index correlation with protein breakdown, dehydration, stress and fatigue. Page 12, line 296, 297: insert ‘statistically significant’ difference Page 12: line 302 – 305: This sentence does not make sense. Do the authors mean that glycogen is the predominant source of energy? Page 12, line 299: The wording should be improved – to prevent fatigue and avoid muscle damage? There is no need to compare every index by detailing the % or –fold increase/decrease. The data, with p values, speaks for itself. Also, If the authors state that there was a X-fold decrease or a % decrease, they should state that this is a decrease in the average values for the groups. Conclusion: Similar studies would now need to be performed in humans.Reviewed by
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This manuscript describes a fairly comprehensive study investigating links between nutrient intake and DNA methylation of a small number of genes in the context of obesity. The authors correctly note that it is an exploratory study with a number of limitations (for example, the assessment of nutrients was only semiquantitative), and that further studies would benefit from additional analyses (for example, a genome-wide approach). Nevertheless, I believe the study will be of interest to readers of Nutrients, and is particularly relevant for a special issue on nutritional epigenetics. My main concern is with the language of the manuscript, in which there are numerous typographical and grammatical errors. For example:  Title should read: "Nutrient intake" or "Intake of nutrients", rather than "Nutrients intake"  Line 25: The first sentence of the abstract should read: "The aim of the present study was to evaluate DNA methylation alterations induced by dietary nutrients in a population of overweight/obese subjects"  Line 29 of the abstract (and elsewhere): The blood clinical parameters should be in lower case unless they are abbreviated (i.e., they should read: "total cholesterol, LDL, HDL, triglycerides, homocysteine").  Line 34 of the abstract (and elsewhere): "We observed positive significant associations" should read: "We observed significant positive associations"  Line 54: "in the last years a great effort" should read: "in recent years a great effort"  Line 55: "Among antioxidants, carotenoids have been shown free radicals scavenger properties..." should read: "Among antioxidants, carotenoids have been shown to act as free radical scavengers..." The manuscript requires extensive editing of the English language and style. If this is attended to, I believe it would be suitable for publication.Reviewed by