Reviewer’ s report
Title: The relationship between preoperative T helper cytokines in the ileal mucosa and the
pathogenesis of pouchitis
Version: 2Date: 09 Apr 2020
Reviewer: Marco Scarpa
Reviewer's report:
This is a potentially interesting study on Th cytokines network as possible predictors of pouchitis
onset after restorative proctocolectomny. The initial hypothesis (the terminal ileum cytokinenetwork at colectomy as predictor of pouchitis) is intriguing and original and the statisticalanalysis is correct. However, the study is heavily flawed by major issues:1. the study analyse 68 patients out of 183 and it is not clear why these patients had a ileal mucosasample taken while the majority had not2. the cytokines were analysed only with RT-PCR which tell us how much mRNA there is but itdoes not take in account any post trascriptional modification and it is easily altered by the differentsampling and stocking technique. ELISA or IHC or Western blot are essential to confirm the realprotein level in teh ileal mucosa3. the cytokine network analysed in this study included 5 cytokines and, in the microarray era,they are pretty few. Moreover, they become just 2 when 3 of them resulted undetectable in thegreat part of the patients. The analysis of mRNA of two cytokines is insufficient to make anyinference on pouchits pathogenesis and on prognosis. In fact, no Pathology Unit will ever providecytokine mRNA levels to the clinicians. They will do with IHC and a prognostic marker should bea IHC marker.
Are the methods appropriate and well described?
If not, please specify what is required in your comments to the authors.
YesDoes the work include the necessary controls?
If not, please specify which controls are required in your comments to the authors.
YesAre the conclusions drawn adequately supported by the data shown?
If not, please explain in your comments to the authors.
NoAre 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 further
assessment in your comments to the editors.
I am able to assess the statisticsQuality 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 financially from the publication of thismanuscript, either now or in 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.
I declare that I have no competing interestsI 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 reportincluding 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 CommonsCC-BY license (http://creativecommons.org/licenses/by/4.0/). I understand that any commentswhich I do not wish to be included in my named report can be included as confidential commentsto the editors, which will not be published.
I agree to the open peer review policy of the journal
Reviewer’ s report
Title: The relationship between preoperative T helper cytokines in the ileal mucosa and the
pathogenesis of pouchitis
Version: 2Date: 09 Apr 2020
Reviewer: Marco Scarpa
Reviewer's report:
This is a potentially interesting study on Th cytokines network as possible predictors of pouchitis
onset after restorative proctocolectomny. The initial hypothesis (the terminal ileum cytokinenetwork at colectomy as predictor of pouchitis) is intriguing and original and the statisticalanalysis is correct. However, the study is heavily flawed by major issues:1. the study analyse 68 patients out of 183 and it is not clear why these patients had a ileal mucosasample taken while the majority had not2. the cytokines were analysed only with RT-PCR which tell us how much mRNA there is but itdoes not take in account any post trascriptional modification and it is easily altered by the differentsampling and stocking technique. ELISA or IHC or Western blot are essential to confirm the realprotein level in teh ileal mucosa3. the cytokine network analysed in this study included 5 cytokines and, in the microarray era,they are pretty few. Moreover, they become just 2 when 3 of them resulted undetectable in thegreat part of the patients. The analysis of mRNA of two cytokines is insufficient to make anyinference on pouchits pathogenesis and on prognosis. In fact, no Pathology Unit will ever providecytokine mRNA levels to the clinicians. They will do with IHC and a prognostic marker should bea IHC marker.
Are the methods appropriate and well described?
If not, please specify what is required in your comments to the authors.
YesDoes the work include the necessary controls?
If not, please specify which controls are required in your comments to the authors.
YesAre the conclusions drawn adequately supported by the data shown?
If not, please explain in your comments to the authors.
NoAre 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 further
assessment in your comments to the editors.
I am able to assess the statisticsQuality 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 financially from the publication of thismanuscript, either now or in 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.
I declare that I have no competing interestsI 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 reportincluding 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 CommonsCC-BY license (http://creativecommons.org/licenses/by/4.0/). I understand that any commentswhich I do not wish to be included in my named report can be included as confidential commentsto 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: The relationship between preoperative T helper cytokines in the ileal mucosa and the
pathogenesis of pouchitis
Version: 2Date: 09 Apr 2020
Reviewer: Marco Scarpa
Reviewer's report:
This is a potentially interesting study on Th cytokines network as possible predictors of pouchitis
onset after restorative proctocolectomny. The initial hypothesis (the terminal ileum cytokinenetwork at colectomy as predictor of pouchitis) is intriguing and original and the statisticalanalysis is correct. However, the study is heavily flawed by major issues:1. the study analyse 68 patients out of 183 and it is not clear why these patients had a ileal mucosasample taken while the majority had not2. the cytokines were analysed only with RT-PCR which tell us how much mRNA there is but itdoes not take in account any post trascriptional modification and it is easily altered by the differentsampling and stocking technique. ELISA or IHC or Western blot are essential to confirm the realprotein level in teh ileal mucosa3. the cytokine network analysed in this study included 5 cytokines and, in the microarray era,they are pretty few. Moreover, they become just 2 when 3 of them resulted undetectable in thegreat part of the patients. The analysis of mRNA of two cytokines is insufficient to make anyinference on pouchits pathogenesis and on prognosis. In fact, no Pathology Unit will ever providecytokine mRNA levels to the clinicians. They will do with IHC and a prognostic marker should bea IHC marker.
Are the methods appropriate and well described?
If not, please specify what is required in your comments to the authors.
YesDoes the work include the necessary controls?
If not, please specify which controls are required in your comments to the authors.
YesAre the conclusions drawn adequately supported by the data shown?
If not, please explain in your comments to the authors.
NoAre 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 further
assessment in your comments to the editors.
I am able to assess the statisticsQuality 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 financially from the publication of thismanuscript, either now or in 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.
I declare that I have no competing interestsI 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 reportincluding 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 CommonsCC-BY license (http://creativecommons.org/licenses/by/4.0/). I understand that any commentswhich I do not wish to be included in my named report can be included as confidential commentsto the editors, which will not be published.
I agree to the open peer review policy of the journal
assistant