Reviewer ’s report  
 
Title: Nerve Growth Factor and Tropomyosin Receptor Kinase A are Increased in the Gastric  
Mucosa of Patients With Functional Dyspepsia  
 
Version: 0 Date: 21 Jun 2019  
 
Reviewer: Cristina Giaroni  
 
Reviewer's report:  
 
This study describes the expression of NGF and its high affinity receptor, TrkA, in the gastric 
mucosa of patients affected by functional dyspepsia, and the possible correlation with enhanced 
NGF -TrkA pathway with some of the symptoms associated with the disease. The study has 
merit. However some points should be addressed before publication:  
Background:  
Line 30: EGCs are major cellular components of myenteric  and submucosal ganglia within the 
ENS. This information is missing;  
Methods:  
-Line 5: a second biopsy was used to perform immunocytochemistry….  
-Line 7: Another  
-For immunohistochemistry, immunofluorescence and western blotting procedures there are no 
indications on the controls specificity assays;  
-Which program was used to measure the NGF, GFAP and TrkA staining density?;  
-There are no specifications on the method used to express western blotting;  
-Since, in the mucosa of FD patients, NGF and TrkA may be  expressed by different populations 
of inflammatory cells double staining with appropriate markers (i.e. eosinophils and mast cell) 
should be performed, to better characterize the possible mucosal sources for the neurotrophin;  
 
Statistical analysis: please  specify the relevant data sets for each applied statistical test  
Results:  
-data indicating the severity of symptom scores to the patients are missing;  
-Lines 13, 29: (Figure 1B). (Line 13) NGF staining density and (line 28) TrkA staining density: 
would  better describe the immunohistochemistry approach;  
Discussion:  
-There is no discussion on how the upregulation of NGF -TrkA pathway in the FD patient 
mucosa may affect afferent sensory neurons for the stomach modulating visceral 
hypersensitivity;  
Minor: th e English form should be revised by a Mother Language speaking colleague  
Abstract  
Line 8: .. and motility disorders…;  
Line 15: .. in gastric mucosa of patients with  FD;  
Line 21: remove square brackets after PDS;  
Line 23: The expression of NGF, TrkA and GF AP was examined;  
Line 40: Changes in the expression of NGF, TrkA and GFAP positively..  
 
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
No 
 
Does the work include the necessary controls?  
If not, please spec ify which controls are required in your comments to the authors.  
 
No 
 
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 wo uld 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:  
 
Needs some language corrections before being published  
 
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 t hat may in any way gain or lose 
financi ally 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 organiz ation 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 th e 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 journal. I understand that my name will be included 
on my report to the authors and, if the manuscript is accepted f or 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 in cluded as confidential comments 
to the editors, which will not be pu blished.  
 
I agree to the open peer review policy of the journal  
Reviewer ’s report  
 
Title: Nerve Growth Factor and Tropomyosin Receptor Kinase A are Increased in the Gastric  
Mucosa of Patients With Functional Dyspepsia  
 
Version: 0 Date: 21 Jun 2019  
 
Reviewer: Cristina Giaroni  
 
Reviewer's report:  
 
This study describes the expression of NGF and its high affinity receptor, TrkA, in the gastric 
mucosa of patients affected by functional dyspepsia, and the possible correlation with enhanced 
NGF -TrkA pathway with some of the symptoms associated with the disease. The study has 
merit. However some points should be addressed before publication:  
Background:  
Line 30: EGCs are major cellular components of myenteric  and submucosal ganglia within the 
ENS. This information is missing;  
Methods:  
-Line 5: a second biopsy was used to perform immunocytochemistry….  
-Line 7: Another  
-For immunohistochemistry, immunofluorescence and western blotting procedures there are no 
indications on the controls specificity assays;  
-Which program was used to measure the NGF, GFAP and TrkA staining density?;  
-There are no specifications on the method used to express western blotting;  
-Since, in the mucosa of FD patients, NGF and TrkA may be  expressed by different populations 
of inflammatory cells double staining with appropriate markers (i.e. eosinophils and mast cell) 
should be performed, to better characterize the possible mucosal sources for the neurotrophin;  
 
Statistical analysis: please  specify the relevant data sets for each applied statistical test  
Results:  
-data indicating the severity of symptom scores to the patients are missing;  
-Lines 13, 29: (Figure 1B). (Line 13) NGF staining density and (line 28) TrkA staining density: 
would  better describe the immunohistochemistry approach;  
Discussion:  
-There is no discussion on how the upregulation of NGF -TrkA pathway in the FD patient 
mucosa may affect afferent sensory neurons for the stomach modulating visceral 
hypersensitivity;  
Minor: th e English form should be revised by a Mother Language speaking colleague  
Abstract  
Line 8: .. and motility disorders…;  
Line 15: .. in gastric mucosa of patients with  FD;  
Line 21: remove square brackets after PDS;  
Line 23: The expression of NGF, TrkA and GF AP was examined;  
Line 40: Changes in the expression of NGF, TrkA and GFAP positively..  
 
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
No 
 
Does the work include the necessary controls?  
If not, please spec ify which controls are required in your comments to the authors.  
 
No 
 
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 wo uld 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:  
 
Needs some language corrections before being published  
 
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 t hat may in any way gain or lose 
financi ally 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 organiz ation 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 th e 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 journal. I understand that my name will be included 
on my report to the authors and, if the manuscript is accepted f or 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 in cluded as confidential comments 
to the editors, which will not be pu blished.  
 
I agree to the open peer review policy of the journal  
Please review this paper.
user
Reviewer ’s report  
 
Title: Nerve Growth Factor and Tropomyosin Receptor Kinase A are Increased in the Gastric  
Mucosa of Patients With Functional Dyspepsia  
 
Version: 0 Date: 21 Jun 2019  
 
Reviewer: Cristina Giaroni  
 
Reviewer's report:  
 
This study describes the expression of NGF and its high affinity receptor, TrkA, in the gastric 
mucosa of patients affected by functional dyspepsia, and the possible correlation with enhanced 
NGF -TrkA pathway with some of the symptoms associated with the disease. The study has 
merit. However some points should be addressed before publication:  
Background:  
Line 30: EGCs are major cellular components of myenteric  and submucosal ganglia within the 
ENS. This information is missing;  
Methods:  
-Line 5: a second biopsy was used to perform immunocytochemistry….  
-Line 7: Another  
-For immunohistochemistry, immunofluorescence and western blotting procedures there are no 
indications on the controls specificity assays;  
-Which program was used to measure the NGF, GFAP and TrkA staining density?;  
-There are no specifications on the method used to express western blotting;  
-Since, in the mucosa of FD patients, NGF and TrkA may be  expressed by different populations 
of inflammatory cells double staining with appropriate markers (i.e. eosinophils and mast cell) 
should be performed, to better characterize the possible mucosal sources for the neurotrophin;  
 
Statistical analysis: please  specify the relevant data sets for each applied statistical test  
Results:  
-data indicating the severity of symptom scores to the patients are missing;  
-Lines 13, 29: (Figure 1B). (Line 13) NGF staining density and (line 28) TrkA staining density: 
would  better describe the immunohistochemistry approach;  
Discussion:  
-There is no discussion on how the upregulation of NGF -TrkA pathway in the FD patient 
mucosa may affect afferent sensory neurons for the stomach modulating visceral 
hypersensitivity;  
Minor: th e English form should be revised by a Mother Language speaking colleague  
Abstract  
Line 8: .. and motility disorders…;  
Line 15: .. in gastric mucosa of patients with  FD;  
Line 21: remove square brackets after PDS;  
Line 23: The expression of NGF, TrkA and GF AP was examined;  
Line 40: Changes in the expression of NGF, TrkA and GFAP positively..  
 
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
No 
 
Does the work include the necessary controls?  
If not, please spec ify which controls are required in your comments to the authors.  
 
No 
 
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 wo uld 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:  
 
Needs some language corrections before being published  
 
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 t hat may in any way gain or lose 
financi ally 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 organiz ation 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 th e 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 journal. I understand that my name will be included 
on my report to the authors and, if the manuscript is accepted f or 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 in cluded as confidential comments 
to the editors, which will not be pu blished.  
 
I agree to the open peer review policy of the journal  
assistant