Reviewer’s report  
Title: Hepcidin Levels Correlate to Iron Parameters, but not St eatohepatitis, in Non -Alcoholic 
Fatty Liver Disease  
 
Version: 0 Date: 16 Apr 2018  
 
Reviewer: Abhishek Agnihotri  
 
Reviewer's report:  
I congratulate the authors on a well written and p ertinent manuscript. Iron dysmetabolism and 
hepcidin in NAFLD has been recently under investigation. There have been some results to 
suggest that managing iron overload could have therapeutic implications in management of 
NAFLD. The association among DIOS,  metabolic syndrome, and NAFLD is being investigated 
as a new risk factor to predict the development of overt cardiovascular and hepatic diseases, and 
possibly hepatocellular carcinoma. Indeed, iron depletion, most frequently achieved by 
phlebotomy, has be en shown to decrease metabolic alterations and liver enzymes in controlled 
studies in NAFLD. Thus I agree with the need to further study the pathogenesis of DIOS. 
Although I admire the exhaustive methodology employed, ideally there could have been a more 
focussed approach towards selecting the cases included in the study and a clear planning 
regarding calculating an adequate sample size.  
 
1) Please define CLD.  
 
2) If this was a prospective study and not a retrospective analysis of prospectively maintained 
database, please explain further if a sample size estimation and control population estimation 
was performed before the start of study.  
 
3) If the goal was to study NAFLD in this prospective study, why were all liver disease included, 
more pertinent would h ave been NAFLD vs other CLD. Inclusion of HH may not have been 
necessary. The aim has to be clarified in introduction.  
 
4) The size of controls relative to total number of patients is a limitation and needs to be 
mentioned in discussion.  
 
5) Please highlight that the biggest limitation is the sample size and also that the study is 
underpowered to make the conclusion stated.  
 
6) Please suggest in discussion a future direction to study DIOS in NAFLD.  
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
Yes 
Does the work include the necessary controls?  
If not, please s pecify which controls are required in your comments to the authors.  
Yes 
Are the conclusions drawn adequately supported by the data shown?  
If not, please explain in your comments to the authors.  
Yes 
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 speci fy what aspects require further 
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:  
Acceptable  
Declaration of competing interests  
Please complete a declaration of competing 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 financial ly from the publication of this 
manuscript, either now or in the future?  
2. Do you hold any stocks or shares in an organisation t hat may in any way gain or lose 
financially from the publication of this manuscript, eith er 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, w rite 'I declare that I have no competing interests' 
below. If your reply is yes to any, please give details below.  
 
none  
 
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 authors' 
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 t he journal  
Reviewer’s report  
Title: Hepcidin Levels Correlate to Iron Parameters, but not St eatohepatitis, in Non -Alcoholic 
Fatty Liver Disease  
 
Version: 0 Date: 16 Apr 2018  
 
Reviewer: Abhishek Agnihotri  
 
Reviewer's report:  
I congratulate the authors on a well written and p ertinent manuscript. Iron dysmetabolism and 
hepcidin in NAFLD has been recently under investigation. There have been some results to 
suggest that managing iron overload could have therapeutic implications in management of 
NAFLD. The association among DIOS,  metabolic syndrome, and NAFLD is being investigated 
as a new risk factor to predict the development of overt cardiovascular and hepatic diseases, and 
possibly hepatocellular carcinoma. Indeed, iron depletion, most frequently achieved by 
phlebotomy, has be en shown to decrease metabolic alterations and liver enzymes in controlled 
studies in NAFLD. Thus I agree with the need to further study the pathogenesis of DIOS. 
Although I admire the exhaustive methodology employed, ideally there could have been a more 
focussed approach towards selecting the cases included in the study and a clear planning 
regarding calculating an adequate sample size.  
 
1) Please define CLD.  
 
2) If this was a prospective study and not a retrospective analysis of prospectively maintained 
database, please explain further if a sample size estimation and control population estimation 
was performed before the start of study.  
 
3) If the goal was to study NAFLD in this prospective study, why were all liver disease included, 
more pertinent would h ave been NAFLD vs other CLD. Inclusion of HH may not have been 
necessary. The aim has to be clarified in introduction.  
 
4) The size of controls relative to total number of patients is a limitation and needs to be 
mentioned in discussion.  
 
5) Please highlight that the biggest limitation is the sample size and also that the study is 
underpowered to make the conclusion stated.  
 
6) Please suggest in discussion a future direction to study DIOS in NAFLD.  
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
Yes 
Does the work include the necessary controls?  
If not, please s pecify which controls are required in your comments to the authors.  
Yes 
Are the conclusions drawn adequately supported by the data shown?  
If not, please explain in your comments to the authors.  
Yes 
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 speci fy what aspects require further 
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:  
Acceptable  
Declaration of competing interests  
Please complete a declaration of competing 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 financial ly from the publication of this 
manuscript, either now or in the future?  
2. Do you hold any stocks or shares in an organisation t hat may in any way gain or lose 
financially from the publication of this manuscript, eith er 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, w rite 'I declare that I have no competing interests' 
below. If your reply is yes to any, please give details below.  
 
none  
 
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 authors' 
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 t he journal  
Please review this paper.
user
Reviewer’s report  
Title: Hepcidin Levels Correlate to Iron Parameters, but not St eatohepatitis, in Non -Alcoholic 
Fatty Liver Disease  
 
Version: 0 Date: 16 Apr 2018  
 
Reviewer: Abhishek Agnihotri  
 
Reviewer's report:  
I congratulate the authors on a well written and p ertinent manuscript. Iron dysmetabolism and 
hepcidin in NAFLD has been recently under investigation. There have been some results to 
suggest that managing iron overload could have therapeutic implications in management of 
NAFLD. The association among DIOS,  metabolic syndrome, and NAFLD is being investigated 
as a new risk factor to predict the development of overt cardiovascular and hepatic diseases, and 
possibly hepatocellular carcinoma. Indeed, iron depletion, most frequently achieved by 
phlebotomy, has be en shown to decrease metabolic alterations and liver enzymes in controlled 
studies in NAFLD. Thus I agree with the need to further study the pathogenesis of DIOS. 
Although I admire the exhaustive methodology employed, ideally there could have been a more 
focussed approach towards selecting the cases included in the study and a clear planning 
regarding calculating an adequate sample size.  
 
1) Please define CLD.  
 
2) If this was a prospective study and not a retrospective analysis of prospectively maintained 
database, please explain further if a sample size estimation and control population estimation 
was performed before the start of study.  
 
3) If the goal was to study NAFLD in this prospective study, why were all liver disease included, 
more pertinent would h ave been NAFLD vs other CLD. Inclusion of HH may not have been 
necessary. The aim has to be clarified in introduction.  
 
4) The size of controls relative to total number of patients is a limitation and needs to be 
mentioned in discussion.  
 
5) Please highlight that the biggest limitation is the sample size and also that the study is 
underpowered to make the conclusion stated.  
 
6) Please suggest in discussion a future direction to study DIOS in NAFLD.  
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
Yes 
Does the work include the necessary controls?  
If not, please s pecify which controls are required in your comments to the authors.  
Yes 
Are the conclusions drawn adequately supported by the data shown?  
If not, please explain in your comments to the authors.  
Yes 
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 speci fy what aspects require further 
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:  
Acceptable  
Declaration of competing interests  
Please complete a declaration of competing 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 financial ly from the publication of this 
manuscript, either now or in the future?  
2. Do you hold any stocks or shares in an organisation t hat may in any way gain or lose 
financially from the publication of this manuscript, eith er 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, w rite 'I declare that I have no competing interests' 
below. If your reply is yes to any, please give details below.  
 
none  
 
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 authors' 
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 t he journal  
assistant