Reviewer ’s report  
Title: A New Method to Induce Nonalcoholic Steatohepatitis (NASH) in Mice  
Version: 2 Date: 28 Feb 2019  
 
Reviewer: Reviewer 2  
Reviewer's report:  
REVISION ASSESSMENT FROM THE ACADEMIC PEER REVIEWER:  
Has the author addressed your concerns sufficiently for you to now recommend the work as a 
technically sound contribution?  
No 
 
Reviewer comments: The study described by the authors sounds but is unfortunately based on a 
non-novel approach and contains limited measurement s. In detail:  
1. Statistically significant results should be described and commented. Non -significant results 
mean the number is probably to low to highlight a difference, or there is no difference at all. A 
trend is not informative.  
2. The paragraph "ef fect of different diets feeding on liver enzymes" is actually only focusing on 
transaminases. Such enzymes are an indirect proof of an hepatic damage/necrosis. Fat diet and 
cigarette smoke are well -know to effect several additional parameters. Functional e nzymes such 
as cytochrome P450 (i.e. 1A2 forms) should be investigated and evaluated. The inducible effect 
on metabolic enzyme by the diets described by the authors are not leading to any conclusion or 
relevant observations. Analysis on hepatic parameters,  including CYP450 enzymes and phase II 
metabolism as well as urea cycle enzymatic activity are highly encouraged.  
3. Histopathological evaluation has been largely described in literature for NASH/NAFLD 
models. Without any additional biomolecular measurements, there is no way to draw any 
important and relevant conclusions that has not been described yet.  
The authors s hould consider a short communication in some experimental journal. In particular, 
due to the fact that the Material and Methods section is so detailed.  
Statistical analysis are not clear (how p values are calculated and why SEM instead of SD for a 
n=8?).  
Finally, the authors should carefully check and editing the English language.  
Minor issue: all the data should have the same amount of digit after the comma to compare. A 
graph (histogram with dot plot and mean+SD) could help the reader to understand and  the 
authors to be more organized during the results description.  
Are the methods appropriate and well described?  
If not, please specify what is requi red in your comments to the authors.  
 
No 
 
Does the work include the necessary controls?  
If not, please specify which controls are required in your comments to the authors.  
 
No 
 
Are the conclusions drawn adequately supported by the data shown?  
If not, plea se explain in your comments to the authors.  
 
No 
 
Are you able to assess any statistics in the manuscript or would you recommend an 
additional statistical review?  
If an additional statistical review is recommended, please specify what aspects require furth er 
assessment in your comments to the editors.  
 
Not relevant to this manuscript  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Needs some language corrections before being published  
 
Declaration of competing interests  
Please complete a declaration of competi ng interests, considering the following questions:  
 
1. Have you in the past five years received reimbursements, fees, funding, or salary from an 
organisation that may in any way gain or lose financially from the publication of this 
manuscript, either now or i n the future?  
 
2. Do you hold any stocks or shares in an organisation that may in any way gain or lose 
financially from the publication of this manuscript, either now or in the future?  
 
3. Do you hold or are you currently applying for any patents relating to the  content of the 
manuscript?  
 
4. Have you received reimbursements, fees, funding, or salary from an organization that 
holds or has applied for patents relating to the content of the manuscript?  
 
5. Do you have any other financial competing interests?  
 
6. Do you have  any non -financial competing interests in relation to this paper?  
 
If you can answer no to all of the above, write 'I declare that I have no competing interests' 
below. If your reply is yes to any, please give details below.  
 
This reviewer has been recruited by a partner organization, Research Square. Reviewers with 
declared or apparent competin g interests are not utilized for these reviews. This reviewer has 
agreed to publication of their comments online under a Creative Commons Attribution License 
attributed to Research Square and was paid a small honorarium for completing the review within 
a specified timeframe. Honoraria for reviews such as this are paid regardless of the reviewer 
recommendation.  
 
I agree to the open peer  review policy of the journal. I understand that my name will be included 
on my report to the authors and, if the manuscript is accepted for publication, my named report 
including any attachments I upload will be posted on the website along with the author s' 
responses. I agree for my report to be made available under an Open Access Creative Commons 
CC-BY license ( http://creativecommons.org/licenses/by/4.0/ ).  I understand that any comments 
which I do not wish to be included in my named report can be included as confidential comments 
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
Reviewer ’s report  
Title: A New Method to Induce Nonalcoholic Steatohepatitis (NASH) in Mice  
Version: 2 Date: 28 Feb 2019  
 
Reviewer: Reviewer 2  
Reviewer's report:  
REVISION ASSESSMENT FROM THE ACADEMIC PEER REVIEWER:  
Has the author addressed your concerns sufficiently for you to now recommend the work as a 
technically sound contribution?  
No 
 
Reviewer comments: The study described by the authors sounds but is unfortunately based on a 
non-novel approach and contains limited measurement s. In detail:  
1. Statistically significant results should be described and commented. Non -significant results 
mean the number is probably to low to highlight a difference, or there is no difference at all. A 
trend is not informative.  
2. The paragraph "ef fect of different diets feeding on liver enzymes" is actually only focusing on 
transaminases. Such enzymes are an indirect proof of an hepatic damage/necrosis. Fat diet and 
cigarette smoke are well -know to effect several additional parameters. Functional e nzymes such 
as cytochrome P450 (i.e. 1A2 forms) should be investigated and evaluated. The inducible effect 
on metabolic enzyme by the diets described by the authors are not leading to any conclusion or 
relevant observations. Analysis on hepatic parameters,  including CYP450 enzymes and phase II 
metabolism as well as urea cycle enzymatic activity are highly encouraged.  
3. Histopathological evaluation has been largely described in literature for NASH/NAFLD 
models. Without any additional biomolecular measurements, there is no way to draw any 
important and relevant conclusions that has not been described yet.  
The authors s hould consider a short communication in some experimental journal. In particular, 
due to the fact that the Material and Methods section is so detailed.  
Statistical analysis are not clear (how p values are calculated and why SEM instead of SD for a 
n=8?).  
Finally, the authors should carefully check and editing the English language.  
Minor issue: all the data should have the same amount of digit after the comma to compare. A 
graph (histogram with dot plot and mean+SD) could help the reader to understand and  the 
authors to be more organized during the results description.  
Are the methods appropriate and well described?  
If not, please specify what is requi red in your comments to the authors.  
 
No 
 
Does the work include the necessary controls?  
If not, please specify which controls are required in your comments to the authors.  
 
No 
 
Are the conclusions drawn adequately supported by the data shown?  
If not, plea se explain in your comments to the authors.  
 
No 
 
Are you able to assess any statistics in the manuscript or would you recommend an 
additional statistical review?  
If an additional statistical review is recommended, please specify what aspects require furth er 
assessment in your comments to the editors.  
 
Not relevant to this manuscript  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Needs some language corrections before being published  
 
Declaration of competing interests  
Please complete a declaration of competi ng interests, considering the following questions:  
 
1. Have you in the past five years received reimbursements, fees, funding, or salary from an 
organisation that may in any way gain or lose financially from the publication of this 
manuscript, either now or i n the future?  
 
2. Do you hold any stocks or shares in an organisation that may in any way gain or lose 
financially from the publication of this manuscript, either now or in the future?  
 
3. Do you hold or are you currently applying for any patents relating to the  content of the 
manuscript?  
 
4. Have you received reimbursements, fees, funding, or salary from an organization that 
holds or has applied for patents relating to the content of the manuscript?  
 
5. Do you have any other financial competing interests?  
 
6. Do you have  any non -financial competing interests in relation to this paper?  
 
If you can answer no to all of the above, write 'I declare that I have no competing interests' 
below. If your reply is yes to any, please give details below.  
 
This reviewer has been recruited by a partner organization, Research Square. Reviewers with 
declared or apparent competin g interests are not utilized for these reviews. This reviewer has 
agreed to publication of their comments online under a Creative Commons Attribution License 
attributed to Research Square and was paid a small honorarium for completing the review within 
a specified timeframe. Honoraria for reviews such as this are paid regardless of the reviewer 
recommendation.  
 
I agree to the open peer  review policy of the journal. I understand that my name will be included 
on my report to the authors and, if the manuscript is accepted for publication, my named report 
including any attachments I upload will be posted on the website along with the author s' 
responses. I agree for my report to be made available under an Open Access Creative Commons 
CC-BY license ( http://creativecommons.org/licenses/by/4.0/ ).  I understand that any comments 
which I do not wish to be included in my named report can be included as confidential comments 
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
Please review this paper.
user
Reviewer ’s report  
Title: A New Method to Induce Nonalcoholic Steatohepatitis (NASH) in Mice  
Version: 2 Date: 28 Feb 2019  
 
Reviewer: Reviewer 2  
Reviewer's report:  
REVISION ASSESSMENT FROM THE ACADEMIC PEER REVIEWER:  
Has the author addressed your concerns sufficiently for you to now recommend the work as a 
technically sound contribution?  
No 
 
Reviewer comments: The study described by the authors sounds but is unfortunately based on a 
non-novel approach and contains limited measurement s. In detail:  
1. Statistically significant results should be described and commented. Non -significant results 
mean the number is probably to low to highlight a difference, or there is no difference at all. A 
trend is not informative.  
2. The paragraph "ef fect of different diets feeding on liver enzymes" is actually only focusing on 
transaminases. Such enzymes are an indirect proof of an hepatic damage/necrosis. Fat diet and 
cigarette smoke are well -know to effect several additional parameters. Functional e nzymes such 
as cytochrome P450 (i.e. 1A2 forms) should be investigated and evaluated. The inducible effect 
on metabolic enzyme by the diets described by the authors are not leading to any conclusion or 
relevant observations. Analysis on hepatic parameters,  including CYP450 enzymes and phase II 
metabolism as well as urea cycle enzymatic activity are highly encouraged.  
3. Histopathological evaluation has been largely described in literature for NASH/NAFLD 
models. Without any additional biomolecular measurements, there is no way to draw any 
important and relevant conclusions that has not been described yet.  
The authors s hould consider a short communication in some experimental journal. In particular, 
due to the fact that the Material and Methods section is so detailed.  
Statistical analysis are not clear (how p values are calculated and why SEM instead of SD for a 
n=8?).  
Finally, the authors should carefully check and editing the English language.  
Minor issue: all the data should have the same amount of digit after the comma to compare. A 
graph (histogram with dot plot and mean+SD) could help the reader to understand and  the 
authors to be more organized during the results description.  
Are the methods appropriate and well described?  
If not, please specify what is requi red in your comments to the authors.  
 
No 
 
Does the work include the necessary controls?  
If not, please specify which controls are required in your comments to the authors.  
 
No 
 
Are the conclusions drawn adequately supported by the data shown?  
If not, plea se explain in your comments to the authors.  
 
No 
 
Are you able to assess any statistics in the manuscript or would you recommend an 
additional statistical review?  
If an additional statistical review is recommended, please specify what aspects require furth er 
assessment in your comments to the editors.  
 
Not relevant to this manuscript  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Needs some language corrections before being published  
 
Declaration of competing interests  
Please complete a declaration of competi ng interests, considering the following questions:  
 
1. Have you in the past five years received reimbursements, fees, funding, or salary from an 
organisation that may in any way gain or lose financially from the publication of this 
manuscript, either now or i n the future?  
 
2. Do you hold any stocks or shares in an organisation that may in any way gain or lose 
financially from the publication of this manuscript, either now or in the future?  
 
3. Do you hold or are you currently applying for any patents relating to the  content of the 
manuscript?  
 
4. Have you received reimbursements, fees, funding, or salary from an organization that 
holds or has applied for patents relating to the content of the manuscript?  
 
5. Do you have any other financial competing interests?  
 
6. Do you have  any non -financial competing interests in relation to this paper?  
 
If you can answer no to all of the above, write 'I declare that I have no competing interests' 
below. If your reply is yes to any, please give details below.  
 
This reviewer has been recruited by a partner organization, Research Square. Reviewers with 
declared or apparent competin g interests are not utilized for these reviews. This reviewer has 
agreed to publication of their comments online under a Creative Commons Attribution License 
attributed to Research Square and was paid a small honorarium for completing the review within 
a specified timeframe. Honoraria for reviews such as this are paid regardless of the reviewer 
recommendation.  
 
I agree to the open peer  review policy of the journal. I understand that my name will be included 
on my report to the authors and, if the manuscript is accepted for publication, my named report 
including any attachments I upload will be posted on the website along with the author s' 
responses. I agree for my report to be made available under an Open Access Creative Commons 
CC-BY license ( http://creativecommons.org/licenses/by/4.0/ ).  I understand that any comments 
which I do not wish to be included in my named report can be included as confidential comments 
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
assistant