Reviewer ’s report  
 
Title: Abnormal ileal stem -, neurogenin3 - and enteroendocrine -cells in patients with irritable 
bowel syndrome  
 
Version: 0 Date: 17 Apr 2017  
 
Reviewer: Martin Martin  
 
Reviewer's report:  
 
This manuscript by El -Salhy and Gilja focuses on an asses sment of ileal staining of intestinal 
strem cells  (ISCs) and enteroendocrine cells (EECs) in a cohort of well characterized adult 
Norwegian patients with various forms of IBS.  
The analysis was limited to an assessment of ISCs stained with Msi -1, EEC prog enitors stained 
with Neurog3, and various differentiated EECs stained with numerous markers including CHGA, 
PYY and other antibodies.  
Using these antibodies, the authors found that IBS was associated with a decline in Msi -1, 
Neurog3, CHGA and serotonin.  
Statistical assessment and numerical data (Fig 1) appears to be appropriate.  
However, staining for Msi -1 appears to be of a transverse section with most of the crypt units 
staining negative for this ISC marker.  
The appropriate sagittal cut would be mor e representative of the crypt base where the ISCs are 
located.  
I would suggest restaining with appropriately sectioned slides.  
I'm also very concerned about the NEUROG3 staining shown in Figure 3.  
NEUROG3 should only be expressed in the crypt base, and not along the villus as is shown in 
this figure.  
Moreover, the B panel gives the appearance of NEUROG3 staining the cytoplasm of the EECs 
like a CHGA antibody, and this suggests that the staining is non-specific.  
The authors should describe how these findings are any different than their other descriptions of 
changes in the EECs of patients with IBS.  
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 specify 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.  
 
Unable to assess  
 
Are you able to assess any st atistics 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 statisti cs 
 
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 y ears 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 in the future?  
 
2. Do you hold any stocks or shares in an organisation that may i n 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, fund ing, 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 interests'  
 
I agree to the open peer review policy of the journ al. 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 m y 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: Abnormal ileal stem -, neurogenin3 - and enteroendocrine -cells in patients with irritable 
bowel syndrome  
 
Version: 0 Date: 17 Apr 2017  
 
Reviewer: Martin Martin  
 
Reviewer's report:  
 
This manuscript by El -Salhy and Gilja focuses on an asses sment of ileal staining of intestinal 
strem cells  (ISCs) and enteroendocrine cells (EECs) in a cohort of well characterized adult 
Norwegian patients with various forms of IBS.  
The analysis was limited to an assessment of ISCs stained with Msi -1, EEC prog enitors stained 
with Neurog3, and various differentiated EECs stained with numerous markers including CHGA, 
PYY and other antibodies.  
Using these antibodies, the authors found that IBS was associated with a decline in Msi -1, 
Neurog3, CHGA and serotonin.  
Statistical assessment and numerical data (Fig 1) appears to be appropriate.  
However, staining for Msi -1 appears to be of a transverse section with most of the crypt units 
staining negative for this ISC marker.  
The appropriate sagittal cut would be mor e representative of the crypt base where the ISCs are 
located.  
I would suggest restaining with appropriately sectioned slides.  
I'm also very concerned about the NEUROG3 staining shown in Figure 3.  
NEUROG3 should only be expressed in the crypt base, and not along the villus as is shown in 
this figure.  
Moreover, the B panel gives the appearance of NEUROG3 staining the cytoplasm of the EECs 
like a CHGA antibody, and this suggests that the staining is non-specific.  
The authors should describe how these findings are any different than their other descriptions of 
changes in the EECs of patients with IBS.  
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 specify 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.  
 
Unable to assess  
 
Are you able to assess any st atistics 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 statisti cs 
 
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 y ears 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 in the future?  
 
2. Do you hold any stocks or shares in an organisation that may i n 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, fund ing, 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 interests'  
 
I agree to the open peer review policy of the journ al. 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 m y 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: Abnormal ileal stem -, neurogenin3 - and enteroendocrine -cells in patients with irritable 
bowel syndrome  
 
Version: 0 Date: 17 Apr 2017  
 
Reviewer: Martin Martin  
 
Reviewer's report:  
 
This manuscript by El -Salhy and Gilja focuses on an asses sment of ileal staining of intestinal 
strem cells  (ISCs) and enteroendocrine cells (EECs) in a cohort of well characterized adult 
Norwegian patients with various forms of IBS.  
The analysis was limited to an assessment of ISCs stained with Msi -1, EEC prog enitors stained 
with Neurog3, and various differentiated EECs stained with numerous markers including CHGA, 
PYY and other antibodies.  
Using these antibodies, the authors found that IBS was associated with a decline in Msi -1, 
Neurog3, CHGA and serotonin.  
Statistical assessment and numerical data (Fig 1) appears to be appropriate.  
However, staining for Msi -1 appears to be of a transverse section with most of the crypt units 
staining negative for this ISC marker.  
The appropriate sagittal cut would be mor e representative of the crypt base where the ISCs are 
located.  
I would suggest restaining with appropriately sectioned slides.  
I'm also very concerned about the NEUROG3 staining shown in Figure 3.  
NEUROG3 should only be expressed in the crypt base, and not along the villus as is shown in 
this figure.  
Moreover, the B panel gives the appearance of NEUROG3 staining the cytoplasm of the EECs 
like a CHGA antibody, and this suggests that the staining is non-specific.  
The authors should describe how these findings are any different than their other descriptions of 
changes in the EECs of patients with IBS.  
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 specify 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.  
 
Unable to assess  
 
Are you able to assess any st atistics 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 statisti cs 
 
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 y ears 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 in the future?  
 
2. Do you hold any stocks or shares in an organisation that may i n 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, fund ing, 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 interests'  
 
I agree to the open peer review policy of the journ al. 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 m y 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