Reviewer ’s report  
Title: A New Method to Induce Nonalcoholic Steatohepatitis (NASH) in Mice  
Version: 1 Date: 21 Jan 2019  
 
Reviewer: Reviewer 2  
Reviewer's report:  
PEER REVIEWER ASSESSMENTS:  
OBJECTIVE - Full research articles:  is there a clear objective that addresses a testable research 
question(s) (brief or other article types: is there a clear objective)?  
No - there are minor issues  
 
DESIGN - Is the current approach (including controls and analysis protocols) appropriate for  the 
objective?  
No - there are minor issues  
 
EXECUTION - Are the experiments and analyses performed with technical rigor to allow 
confidence in the results?  
Yes - experiments and analyses were performed appropriately  
 
Statistics - Is the use of statistics in the manuscript appropriate?  
Yes - appropriate statistical analyses have been used in the study  
 
INTERPRETATION - Is the current interpretation/discussion of the results reasonable and not 
overstated?  
No - there are major issues  
 
OVERALL MANUSCRIPT POTENTIAL - Is the current version of this work technically 
sound? If not, can revisions be made to make the work technically sound?  
Maybe - with major revisions  
 
PEER REVIEWER COMMENTS:  
GENERAL COMMENTS: The study described by the aut hors is well designed but based on a 
non-novel approach and contains limited and non -critical measurements. The authors mentioned 
"an ideal disease model" for NASH, but the model used in the study is not novel and less than 
ideal. Moreover, additional anal ysis (biomolecular or immunohistological measurements) on all 
the groups are strongly encouraged. In the current manuscript, extremely long description for 
results with limited or null statistical importance have been provided; long description of the 
meth odological approaches with large repetitions (last 5 lines at page 5 and the next 9 lines in 
page 6 are a repetition of page 5); figures containing histograms with dot plots may be more 
informative to highlight data distribution; additional analysis on mor e hepatic parameters, 
including phase I (CYP450 enzymes) and phase II metabolism and urea cycle are highly 
important for the study.  
 
REQUESTED REVISIONS:  
Firstly, it is not clear what the authors aimed to prove or investigate. The generation of an "ideal 
disease model" for NASH disease appears to be one of the aims (as stated at page 3), but the 
animals exposed to different diets are not new and their relevance is questionable. The 
parameters measured and analyzed are not sufficient and poorly informative.  additional analyses 
are encouraged with more specific and punctual measurements based on different diets/smoking 
effects. Finally, the authors should focus their Conclusion on data obtained during the study, 
limiting comments and discussion on previous wo rk. Negative results, as "fructose consumption 
was not able to change significantly the studied biochemical parameters" (as stated at page 13), 
may be of importance, but only if the analyzed parameters are of relevance and specifically 
targeted.  
 
ADDITIONAL  REQUESTS/SUGGESTIONS:  
English language editing is recommended.  
Note: This reviewer report can be downloaded - see attached pdf file.  
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.  
 
I am able to assess the statistics  
 
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: 1 Date: 21 Jan 2019  
 
Reviewer: Reviewer 2  
Reviewer's report:  
PEER REVIEWER ASSESSMENTS:  
OBJECTIVE - Full research articles:  is there a clear objective that addresses a testable research 
question(s) (brief or other article types: is there a clear objective)?  
No - there are minor issues  
 
DESIGN - Is the current approach (including controls and analysis protocols) appropriate for  the 
objective?  
No - there are minor issues  
 
EXECUTION - Are the experiments and analyses performed with technical rigor to allow 
confidence in the results?  
Yes - experiments and analyses were performed appropriately  
 
Statistics - Is the use of statistics in the manuscript appropriate?  
Yes - appropriate statistical analyses have been used in the study  
 
INTERPRETATION - Is the current interpretation/discussion of the results reasonable and not 
overstated?  
No - there are major issues  
 
OVERALL MANUSCRIPT POTENTIAL - Is the current version of this work technically 
sound? If not, can revisions be made to make the work technically sound?  
Maybe - with major revisions  
 
PEER REVIEWER COMMENTS:  
GENERAL COMMENTS: The study described by the aut hors is well designed but based on a 
non-novel approach and contains limited and non -critical measurements. The authors mentioned 
"an ideal disease model" for NASH, but the model used in the study is not novel and less than 
ideal. Moreover, additional anal ysis (biomolecular or immunohistological measurements) on all 
the groups are strongly encouraged. In the current manuscript, extremely long description for 
results with limited or null statistical importance have been provided; long description of the 
meth odological approaches with large repetitions (last 5 lines at page 5 and the next 9 lines in 
page 6 are a repetition of page 5); figures containing histograms with dot plots may be more 
informative to highlight data distribution; additional analysis on mor e hepatic parameters, 
including phase I (CYP450 enzymes) and phase II metabolism and urea cycle are highly 
important for the study.  
 
REQUESTED REVISIONS:  
Firstly, it is not clear what the authors aimed to prove or investigate. The generation of an "ideal 
disease model" for NASH disease appears to be one of the aims (as stated at page 3), but the 
animals exposed to different diets are not new and their relevance is questionable. The 
parameters measured and analyzed are not sufficient and poorly informative.  additional analyses 
are encouraged with more specific and punctual measurements based on different diets/smoking 
effects. Finally, the authors should focus their Conclusion on data obtained during the study, 
limiting comments and discussion on previous wo rk. Negative results, as "fructose consumption 
was not able to change significantly the studied biochemical parameters" (as stated at page 13), 
may be of importance, but only if the analyzed parameters are of relevance and specifically 
targeted.  
 
ADDITIONAL  REQUESTS/SUGGESTIONS:  
English language editing is recommended.  
Note: This reviewer report can be downloaded - see attached pdf file.  
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.  
 
I am able to assess the statistics  
 
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: 1 Date: 21 Jan 2019  
 
Reviewer: Reviewer 2  
Reviewer's report:  
PEER REVIEWER ASSESSMENTS:  
OBJECTIVE - Full research articles:  is there a clear objective that addresses a testable research 
question(s) (brief or other article types: is there a clear objective)?  
No - there are minor issues  
 
DESIGN - Is the current approach (including controls and analysis protocols) appropriate for  the 
objective?  
No - there are minor issues  
 
EXECUTION - Are the experiments and analyses performed with technical rigor to allow 
confidence in the results?  
Yes - experiments and analyses were performed appropriately  
 
Statistics - Is the use of statistics in the manuscript appropriate?  
Yes - appropriate statistical analyses have been used in the study  
 
INTERPRETATION - Is the current interpretation/discussion of the results reasonable and not 
overstated?  
No - there are major issues  
 
OVERALL MANUSCRIPT POTENTIAL - Is the current version of this work technically 
sound? If not, can revisions be made to make the work technically sound?  
Maybe - with major revisions  
 
PEER REVIEWER COMMENTS:  
GENERAL COMMENTS: The study described by the aut hors is well designed but based on a 
non-novel approach and contains limited and non -critical measurements. The authors mentioned 
"an ideal disease model" for NASH, but the model used in the study is not novel and less than 
ideal. Moreover, additional anal ysis (biomolecular or immunohistological measurements) on all 
the groups are strongly encouraged. In the current manuscript, extremely long description for 
results with limited or null statistical importance have been provided; long description of the 
meth odological approaches with large repetitions (last 5 lines at page 5 and the next 9 lines in 
page 6 are a repetition of page 5); figures containing histograms with dot plots may be more 
informative to highlight data distribution; additional analysis on mor e hepatic parameters, 
including phase I (CYP450 enzymes) and phase II metabolism and urea cycle are highly 
important for the study.  
 
REQUESTED REVISIONS:  
Firstly, it is not clear what the authors aimed to prove or investigate. The generation of an "ideal 
disease model" for NASH disease appears to be one of the aims (as stated at page 3), but the 
animals exposed to different diets are not new and their relevance is questionable. The 
parameters measured and analyzed are not sufficient and poorly informative.  additional analyses 
are encouraged with more specific and punctual measurements based on different diets/smoking 
effects. Finally, the authors should focus their Conclusion on data obtained during the study, 
limiting comments and discussion on previous wo rk. Negative results, as "fructose consumption 
was not able to change significantly the studied biochemical parameters" (as stated at page 13), 
may be of importance, but only if the analyzed parameters are of relevance and specifically 
targeted.  
 
ADDITIONAL  REQUESTS/SUGGESTIONS:  
English language editing is recommended.  
Note: This reviewer report can be downloaded - see attached pdf file.  
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.  
 
I am able to assess the statistics  
 
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