Reviewer ’s report  
 
Title: Histopathological features of the proper gastric glands in FVB/N -background mice 
carrying constitutively -active aryl -hydrocarbon receptor  
 
Version: 1 Date: 04 Jul 2018  
 
Reviewer: Robert Coffey  
 
Reviewer's report:  
 
Dantsuka et al. report that expression of constitutively active AhR in FVB/N mice causes gastric 
lesions mimicking gastritis cystica profunda earlier than in C57BL/6 mice. This revised 
manuscript also shows dilated glands contain SPEM metaplasia by TFF2 immunostaining. The 
findings are novel; however, additional work is ne eded to solidify their findings.  
 
Major points  
1. The authors used single TFF2 immunostaining to show SPEM. However, it requires dual 
staining of neck cell markers (TFF2 or GS2) and chief cell markers (GIF or pepsinogen) to call it 
SPEM. Alternatively, CD44 v9 single staining can be used. In addition, TFF2 staining is not 
convincing because control stomach (Fig. 6l) does not show TFF2 positivity in neck cells.  
2. In this reviewer's opinion, the two major pathological findings in the CA -AhR mouse 
stomachs are SPEM and gastritis cystica profunda. Those two pathologies have been previously 
associated in different mouse models (Roepke TK et al. PLoS One. 2010; 5(7): e11451). SPEM 
arises in the background of oxyntic atrophy (loss of parietal cells). Also, movement of 
proliferation center to the base of the glands is one of the characteristics of SPEM. This reviewer 
recommends that the authors focus on these two pathologies.  
 
Minor points  
1. In line 208, the authors argue that these cysts are lined by simple squamous  or columnar 
epithelial cells. But Fig. 2a does not show any squamous epithelium. They look like gastric 
epithelium with area of flattening.  
2. Terminology, lamina muscularis is still used in the abstract. It should be changed to (lamina) 
muscularis mucosae.  
3. The authors describe that the proliferating cells shifted from the isthmus to the fundus. Fundus 
sounds confusing because it is used as  one of the regions of the stomach. The base of the glands 
can be used instead of the fundus.  
4. Legend of Fig. 1b - it is not clear that it is a control stomach.  
5. Fig. 1f is a picture of empty stomach while other stomachs in the Fig. 1 are stomachs 
containing food. It would be better to be consistent.  
6. In line 317, the authors stated "Both dysplasia and EM cysts". There is no data showing 
dysplasia in the manuscript. Presumably, the authors meant metaplasia.  
7. In line 331, the sentence "SPEM chief cel ls to transdifferentiate into mucous cell metaplasia" 
is confusing because it is thought that chief cells transdifferentiate into SEPM metaplastic cells.  
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
Yes 
 
Does the work inc lude 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.  
 
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 specify what aspects require further  
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:  
 
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 this  
manuscript, 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 com peting 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 jou rnal. 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 the journal  
Reviewer ’s report  
 
Title: Histopathological features of the proper gastric glands in FVB/N -background mice 
carrying constitutively -active aryl -hydrocarbon receptor  
 
Version: 1 Date: 04 Jul 2018  
 
Reviewer: Robert Coffey  
 
Reviewer's report:  
 
Dantsuka et al. report that expression of constitutively active AhR in FVB/N mice causes gastric 
lesions mimicking gastritis cystica profunda earlier than in C57BL/6 mice. This revised 
manuscript also shows dilated glands contain SPEM metaplasia by TFF2 immunostaining. The 
findings are novel; however, additional work is ne eded to solidify their findings.  
 
Major points  
1. The authors used single TFF2 immunostaining to show SPEM. However, it requires dual 
staining of neck cell markers (TFF2 or GS2) and chief cell markers (GIF or pepsinogen) to call it 
SPEM. Alternatively, CD44 v9 single staining can be used. In addition, TFF2 staining is not 
convincing because control stomach (Fig. 6l) does not show TFF2 positivity in neck cells.  
2. In this reviewer's opinion, the two major pathological findings in the CA -AhR mouse 
stomachs are SPEM and gastritis cystica profunda. Those two pathologies have been previously 
associated in different mouse models (Roepke TK et al. PLoS One. 2010; 5(7): e11451). SPEM 
arises in the background of oxyntic atrophy (loss of parietal cells). Also, movement of 
proliferation center to the base of the glands is one of the characteristics of SPEM. This reviewer 
recommends that the authors focus on these two pathologies.  
 
Minor points  
1. In line 208, the authors argue that these cysts are lined by simple squamous  or columnar 
epithelial cells. But Fig. 2a does not show any squamous epithelium. They look like gastric 
epithelium with area of flattening.  
2. Terminology, lamina muscularis is still used in the abstract. It should be changed to (lamina) 
muscularis mucosae.  
3. The authors describe that the proliferating cells shifted from the isthmus to the fundus. Fundus 
sounds confusing because it is used as  one of the regions of the stomach. The base of the glands 
can be used instead of the fundus.  
4. Legend of Fig. 1b - it is not clear that it is a control stomach.  
5. Fig. 1f is a picture of empty stomach while other stomachs in the Fig. 1 are stomachs 
containing food. It would be better to be consistent.  
6. In line 317, the authors stated "Both dysplasia and EM cysts". There is no data showing 
dysplasia in the manuscript. Presumably, the authors meant metaplasia.  
7. In line 331, the sentence "SPEM chief cel ls to transdifferentiate into mucous cell metaplasia" 
is confusing because it is thought that chief cells transdifferentiate into SEPM metaplastic cells.  
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
Yes 
 
Does the work inc lude 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.  
 
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 specify what aspects require further  
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:  
 
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 this  
manuscript, 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 com peting 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 jou rnal. 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 the journal  
Please review this paper.
user
Reviewer ’s report  
 
Title: Histopathological features of the proper gastric glands in FVB/N -background mice 
carrying constitutively -active aryl -hydrocarbon receptor  
 
Version: 1 Date: 04 Jul 2018  
 
Reviewer: Robert Coffey  
 
Reviewer's report:  
 
Dantsuka et al. report that expression of constitutively active AhR in FVB/N mice causes gastric 
lesions mimicking gastritis cystica profunda earlier than in C57BL/6 mice. This revised 
manuscript also shows dilated glands contain SPEM metaplasia by TFF2 immunostaining. The 
findings are novel; however, additional work is ne eded to solidify their findings.  
 
Major points  
1. The authors used single TFF2 immunostaining to show SPEM. However, it requires dual 
staining of neck cell markers (TFF2 or GS2) and chief cell markers (GIF or pepsinogen) to call it 
SPEM. Alternatively, CD44 v9 single staining can be used. In addition, TFF2 staining is not 
convincing because control stomach (Fig. 6l) does not show TFF2 positivity in neck cells.  
2. In this reviewer's opinion, the two major pathological findings in the CA -AhR mouse 
stomachs are SPEM and gastritis cystica profunda. Those two pathologies have been previously 
associated in different mouse models (Roepke TK et al. PLoS One. 2010; 5(7): e11451). SPEM 
arises in the background of oxyntic atrophy (loss of parietal cells). Also, movement of 
proliferation center to the base of the glands is one of the characteristics of SPEM. This reviewer 
recommends that the authors focus on these two pathologies.  
 
Minor points  
1. In line 208, the authors argue that these cysts are lined by simple squamous  or columnar 
epithelial cells. But Fig. 2a does not show any squamous epithelium. They look like gastric 
epithelium with area of flattening.  
2. Terminology, lamina muscularis is still used in the abstract. It should be changed to (lamina) 
muscularis mucosae.  
3. The authors describe that the proliferating cells shifted from the isthmus to the fundus. Fundus 
sounds confusing because it is used as  one of the regions of the stomach. The base of the glands 
can be used instead of the fundus.  
4. Legend of Fig. 1b - it is not clear that it is a control stomach.  
5. Fig. 1f is a picture of empty stomach while other stomachs in the Fig. 1 are stomachs 
containing food. It would be better to be consistent.  
6. In line 317, the authors stated "Both dysplasia and EM cysts". There is no data showing 
dysplasia in the manuscript. Presumably, the authors meant metaplasia.  
7. In line 331, the sentence "SPEM chief cel ls to transdifferentiate into mucous cell metaplasia" 
is confusing because it is thought that chief cells transdifferentiate into SEPM metaplastic cells.  
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
Yes 
 
Does the work inc lude 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.  
 
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 specify what aspects require further  
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:  
 
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 this  
manuscript, 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 com peting 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 jou rnal. 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 the journal  
assistant