Reviewer’s report
Title: 
May chronic cough in chronic obstructive pulmonary disease be a
contraindication of Percutaneous Endoscopic Gastrostomy placement: a case
report
Version: 
5 
Date: 
15 Oct 2020
Reviewer's report:
PEER REVIEWER ASSESSMENTS: 
OBJECTIVE - Full research articles: is there a clear objective that addresses one
or several testable research questions? (Brief or other article types: is there a
clear objective?)
Yes - there is a clear objective
DESIGN - Is the current approach (including controls and analysis protocols)
appropriate for the objective?
Yes - the approach is appropriate
EXECUTION - Are the experiments and analyses performed with sufficient
technical rigor to allow confidence in the results?
No - there are minor issues
STATISTICS - Is the use of statistics in the manuscript appropriate?
N/A - there are no statistics in this study
INTERPRETATION - Is the current interpretation/discussion of the results
reasonable and not overstated?
No - there are minor issues
OVERALL MANUSCRIPT POTENTIAL - Has the author addressed your
concerns sufficiently for you to now recommend the work as a technically sound
contribution? If not, can further revisions be made to make the work technically
sound?
Probably - with minor revisions
PEER REVIEWER COMMENTS: 
GENERAL COMMENTS: Please note I have done this review as a "re-review"
and have highlighted the points of a re-review; 
I did not do the first review and
this evaluation will still address the points that are important. 
HAVE THE AUTHORS RESPONDED ADEQUATELY TO EACH OF THE
QUESTIONS YOU RAISED IN YOUR COMMENTS 
AND WHAT HAVE
THE AUTHORS DONE WELL
The authors have done an excellent job responding to the comprehensive past 3
reviews! 
Well done authors!
HAVE THEY MADE THE APPROPRIATE CHANGES IN THE
MANUSCRIPT 
The authors have made valuable changes that have increased the knowledge of
the reader with VERY useful additional discussion
WHAT IS YOUR OVERALL IMPRESSION OF THE REVISED
MANUSCRIPT 
There is a definite message here but quite frankly I think the message is two-fold.
1. Do not pull the PEG tube up against the abdominal wall too tightly
2. Appropriate tube care is essential
IN WHAT WAYS DOES IT NOT MEET BEST PRACTICE
This revision DOES MEET BEST PRACTOICE
REQUESTED REVISIONS: 
ARE THERE ANY ADDITIONAL REVISIONS THE AUTHORS SHOULD
MAKE
Let me start by saying that I am not one of the original reviewers thus some of
my comments will be new to the authors
1. Abstract line 13 change " dislodgement" to " erosion" 
2. Abstract 
Lines 18-19 reword to "We report the development of BBS in a man
with chronic obstructive pulmonary disease who had 
a persistent chronic
morning cough and required placement of a PEG tube for continuous infusion of
Levodopa/carbidopa intestinal gel 
for advanced Parkinson's disease. 
3 abstract line 25 
add the words " an absolute " 
after the word " representing" 
4. Abstract line 27 add the phrase "regimen of " after the word "appropriate "
5. In the future please number the pages or use a continuous numbering of lines.
All further suggestions will be according to pages with the abstract as page 1 ( as
a reviewer I do not see the title page)
6. page 2 line 14 add the words " and enteral access" after the word nutrition". In
addition 
start the next sentence with "In addition, the PEG tube 
can be used to
intubate the proximal jejunum with an extension tube (J-PEG) to allow the
pharmacological treatment *****"
7. I would not use the abbreviation ADT; it is not a common one
8. Page 2 line 21 start a new paragraph
9. Page 2 line 26. Which part of the bowel? I think you mean "colon during
insertion"
10. Page 2 line 30 change "migrates" to "erodes"
11. Page 2 line 36 what do you mean by leakage of fluid "in the peristomal site"?
Don't you mean "leakage of gastric content from around the PEG tube insertion
site or into the peritoneum"
12. Page 2 line 36 what do you mean about immobility of the PEG tube- You
need to be more clear. Similarly on line 37 " inability to insert. WHAT? The
tube?, fluid? 
13. Page 2 line 52 change to "consent"
14. Page 3 line10 change "routinely" to "routine" and delete 
"control" and
replace by "post insertion endoscopic confirmation of satisfactory PEG
placement"
15 I am not impressed by figures 1 and 2 moreover they should be figure 1A and
B; but I would 
delete them
16. Page 3 line 21 change the word "stoma" to "site" 
And add at the end " and "with visualization of only the J-tube extension" 
17. Page 3 line 10 here is where you should define the type of PEG tube you
used and again on line 30. 
After which delete lines 36-37
18. Page 3 lines 32-35. How does this second situation differ from the first one ?
They seem similar so why not try a needle knife-assisted approach? If the
findings are different please explain better
19. Page 3 lines 48-49. I will have to disagree that the morning cough was not
responsible 
rather the tube was positioned with too much tension placed on the
bumpers and the real problem was the lack of appropriate tube management "
sorry 
if the patient had a persistent cough 
all the time then I might relate it to
coughing, but you say that this was only a morning cough. Therefore I 
cannot
believe it was the cough that did this
20. Page 3 lines 57-59 I do not think this discussion is appropriate. There was too
much traction on the tube and may be that the coughing exacerbated it, but it was
not the coughing that caused this
21. Page 3 line 57. The word "decubitus" is not appropriate 
22. Page 4 lines 6-10 you need to define these types not everyone will have
ready access to the 
internet to review these papers
23. Page 4 lines 13 why are pharyngeal and /or esophageal obstruction a
contraindication? 
That makes no sense to me -indeed obstruction on these
regions is why a PEG may be needed 
24. Page 4 lines 26-28 I do not see how either a radiologic or operative approach
will prevent the care taker of pulling the tube up to the abdominal wall too
tightly. You need to describe how an operative approach will prevent this
25. Page 4 line 32 add to the end of the sentence "as well as to ability to feed
directly into the jejunum"
26. Page 4 line 38 change into and outside of the stomach" to "advanced further
in to the stomach and them pulled back until set to the correct length and
tension"
27 page 4 line 40 change to "the tube needs to have some slack to
accommodate***
28. Page 4 lines 47-49. This explanation makes no sense - the J tube does not
erode the balloon
29. Page 4 line 53 again change migration to erosion 
30. Sorry for all the suggestions, but fixing these will make the report much
clearer
ADDITIONAL REQUESTS/SUGGESTIONS: 
see bove
Publons Reviewer Recognition
. Springer Nature can send verification of this
review directly to Publons (a subsidiary of Clarivate Analytics). If you would
like to take advantage of this service, please click on the “Yes” option below.
Your name, email address, title of the reviewed manuscript, name of the journal,
and date of your review submission (the “Review Data”) will then be transmitted
to Publons upon publication of the manuscript. If you have already registered at
Publons, they will notify you of the receipt of this review and update your profile
as per your settings and their policy. If you are not registered with Publons, you
will receive an email from them asking you to register in order for them to be
able to recognize your review on your new profile page. Publons may use the
Review Data to generate derivative metadata for the benefit of Publons and you
as a reviewer, carefully considering the sensitivity of such information. For
example, Publons may verify your record as a reviewer by updating your profile
published on its webservice if you have registered for such service or help
editors to identify candidate reviewers. Please find the details of processing in
Publons’ privacy policy 
https://publons.com/about/terms
No
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 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.
This reviewer has been recruited by a partner organization, Research Square.
Reviewers with declared or apparent competing interests are not utilized for
these reviews. This reviewer has agreed to publication of their comments online
under a Creative Commons Attribution License attributed to Research Square
and was paid a small honorarium for completing the review within a specified
timeframe. Honoraria for reviews such as this are paid regardless of the reviewer
recommendation.
Reviewer Publication Consent.
 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
) if this manuscript is accepted for
publication. Any comments that I do not wish to be included in the published
report have been included as confidential comments to the editor, which will not
be published.
If you are not happy for us to publish this report, please contact the editorial
office before completing the review.
If you wish, you can include your name in your published report. Please note
you must decide whether to include your name at the start of the process and
confirm this decision whenever prompted. If you change your decision later, we
will not be able to publish your name.
I agree to the terms of the CC-BY 4.0 license; please publish my name with my
report.
Is the study design appropriate to answer the research question (including the use
of appropriate controls), and are the conclusions supported by the evidence
presented?
Yes
Are the methods sufficiently described to allow the study to be repeated?
Yes
Is the use of statistics and treatment of uncertainties appropriate?
No
Is the presentation of the work clear?
Yes
Are the images in this manuscript (including electrophoretic gels and blots) free
from apparent manipulation?
Yes
Reviewer’s report
Title: 
May chronic cough in chronic obstructive pulmonary disease be a
contraindication of Percutaneous Endoscopic Gastrostomy placement: a case
report
Version: 
5 
Date: 
15 Oct 2020
Reviewer's report:
PEER REVIEWER ASSESSMENTS: 
OBJECTIVE - Full research articles: is there a clear objective that addresses one
or several testable research questions? (Brief or other article types: is there a
clear objective?)
Yes - there is a clear objective
DESIGN - Is the current approach (including controls and analysis protocols)
appropriate for the objective?
Yes - the approach is appropriate
EXECUTION - Are the experiments and analyses performed with sufficient
technical rigor to allow confidence in the results?
No - there are minor issues
STATISTICS - Is the use of statistics in the manuscript appropriate?
N/A - there are no statistics in this study
INTERPRETATION - Is the current interpretation/discussion of the results
reasonable and not overstated?
No - there are minor issues
OVERALL MANUSCRIPT POTENTIAL - Has the author addressed your
concerns sufficiently for you to now recommend the work as a technically sound
contribution? If not, can further revisions be made to make the work technically
sound?
Probably - with minor revisions
PEER REVIEWER COMMENTS: 
GENERAL COMMENTS: Please note I have done this review as a "re-review"
and have highlighted the points of a re-review; 
I did not do the first review and
this evaluation will still address the points that are important. 
HAVE THE AUTHORS RESPONDED ADEQUATELY TO EACH OF THE
QUESTIONS YOU RAISED IN YOUR COMMENTS 
AND WHAT HAVE
THE AUTHORS DONE WELL
The authors have done an excellent job responding to the comprehensive past 3
reviews! 
Well done authors!
HAVE THEY MADE THE APPROPRIATE CHANGES IN THE
MANUSCRIPT 
The authors have made valuable changes that have increased the knowledge of
the reader with VERY useful additional discussion
WHAT IS YOUR OVERALL IMPRESSION OF THE REVISED
MANUSCRIPT 
There is a definite message here but quite frankly I think the message is two-fold.
1. Do not pull the PEG tube up against the abdominal wall too tightly
2. Appropriate tube care is essential
IN WHAT WAYS DOES IT NOT MEET BEST PRACTICE
This revision DOES MEET BEST PRACTOICE
REQUESTED REVISIONS: 
ARE THERE ANY ADDITIONAL REVISIONS THE AUTHORS SHOULD
MAKE
Let me start by saying that I am not one of the original reviewers thus some of
my comments will be new to the authors
1. Abstract line 13 change " dislodgement" to " erosion" 
2. Abstract 
Lines 18-19 reword to "We report the development of BBS in a man
with chronic obstructive pulmonary disease who had 
a persistent chronic
morning cough and required placement of a PEG tube for continuous infusion of
Levodopa/carbidopa intestinal gel 
for advanced Parkinson's disease. 
3 abstract line 25 
add the words " an absolute " 
after the word " representing" 
4. Abstract line 27 add the phrase "regimen of " after the word "appropriate "
5. In the future please number the pages or use a continuous numbering of lines.
All further suggestions will be according to pages with the abstract as page 1 ( as
a reviewer I do not see the title page)
6. page 2 line 14 add the words " and enteral access" after the word nutrition". In
addition 
start the next sentence with "In addition, the PEG tube 
can be used to
intubate the proximal jejunum with an extension tube (J-PEG) to allow the
pharmacological treatment *****"
7. I would not use the abbreviation ADT; it is not a common one
8. Page 2 line 21 start a new paragraph
9. Page 2 line 26. Which part of the bowel? I think you mean "colon during
insertion"
10. Page 2 line 30 change "migrates" to "erodes"
11. Page 2 line 36 what do you mean by leakage of fluid "in the peristomal site"?
Don't you mean "leakage of gastric content from around the PEG tube insertion
site or into the peritoneum"
12. Page 2 line 36 what do you mean about immobility of the PEG tube- You
need to be more clear. Similarly on line 37 " inability to insert. WHAT? The
tube?, fluid? 
13. Page 2 line 52 change to "consent"
14. Page 3 line10 change "routinely" to "routine" and delete 
"control" and
replace by "post insertion endoscopic confirmation of satisfactory PEG
placement"
15 I am not impressed by figures 1 and 2 moreover they should be figure 1A and
B; but I would 
delete them
16. Page 3 line 21 change the word "stoma" to "site" 
And add at the end " and "with visualization of only the J-tube extension" 
17. Page 3 line 10 here is where you should define the type of PEG tube you
used and again on line 30. 
After which delete lines 36-37
18. Page 3 lines 32-35. How does this second situation differ from the first one ?
They seem similar so why not try a needle knife-assisted approach? If the
findings are different please explain better
19. Page 3 lines 48-49. I will have to disagree that the morning cough was not
responsible 
rather the tube was positioned with too much tension placed on the
bumpers and the real problem was the lack of appropriate tube management "
sorry 
if the patient had a persistent cough 
all the time then I might relate it to
coughing, but you say that this was only a morning cough. Therefore I 
cannot
believe it was the cough that did this
20. Page 3 lines 57-59 I do not think this discussion is appropriate. There was too
much traction on the tube and may be that the coughing exacerbated it, but it was
not the coughing that caused this
21. Page 3 line 57. The word "decubitus" is not appropriate 
22. Page 4 lines 6-10 you need to define these types not everyone will have
ready access to the 
internet to review these papers
23. Page 4 lines 13 why are pharyngeal and /or esophageal obstruction a
contraindication? 
That makes no sense to me -indeed obstruction on these
regions is why a PEG may be needed 
24. Page 4 lines 26-28 I do not see how either a radiologic or operative approach
will prevent the care taker of pulling the tube up to the abdominal wall too
tightly. You need to describe how an operative approach will prevent this
25. Page 4 line 32 add to the end of the sentence "as well as to ability to feed
directly into the jejunum"
26. Page 4 line 38 change into and outside of the stomach" to "advanced further
in to the stomach and them pulled back until set to the correct length and
tension"
27 page 4 line 40 change to "the tube needs to have some slack to
accommodate***
28. Page 4 lines 47-49. This explanation makes no sense - the J tube does not
erode the balloon
29. Page 4 line 53 again change migration to erosion 
30. Sorry for all the suggestions, but fixing these will make the report much
clearer
ADDITIONAL REQUESTS/SUGGESTIONS: 
see bove
Publons Reviewer Recognition
. Springer Nature can send verification of this
review directly to Publons (a subsidiary of Clarivate Analytics). If you would
like to take advantage of this service, please click on the “Yes” option below.
Your name, email address, title of the reviewed manuscript, name of the journal,
and date of your review submission (the “Review Data”) will then be transmitted
to Publons upon publication of the manuscript. If you have already registered at
Publons, they will notify you of the receipt of this review and update your profile
as per your settings and their policy. If you are not registered with Publons, you
will receive an email from them asking you to register in order for them to be
able to recognize your review on your new profile page. Publons may use the
Review Data to generate derivative metadata for the benefit of Publons and you
as a reviewer, carefully considering the sensitivity of such information. For
example, Publons may verify your record as a reviewer by updating your profile
published on its webservice if you have registered for such service or help
editors to identify candidate reviewers. Please find the details of processing in
Publons’ privacy policy 
https://publons.com/about/terms
No
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 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.
This reviewer has been recruited by a partner organization, Research Square.
Reviewers with declared or apparent competing interests are not utilized for
these reviews. This reviewer has agreed to publication of their comments online
under a Creative Commons Attribution License attributed to Research Square
and was paid a small honorarium for completing the review within a specified
timeframe. Honoraria for reviews such as this are paid regardless of the reviewer
recommendation.
Reviewer Publication Consent.
 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
) if this manuscript is accepted for
publication. Any comments that I do not wish to be included in the published
report have been included as confidential comments to the editor, which will not
be published.
If you are not happy for us to publish this report, please contact the editorial
office before completing the review.
If you wish, you can include your name in your published report. Please note
you must decide whether to include your name at the start of the process and
confirm this decision whenever prompted. If you change your decision later, we
will not be able to publish your name.
I agree to the terms of the CC-BY 4.0 license; please publish my name with my
report.
Is the study design appropriate to answer the research question (including the use
of appropriate controls), and are the conclusions supported by the evidence
presented?
Yes
Are the methods sufficiently described to allow the study to be repeated?
Yes
Is the use of statistics and treatment of uncertainties appropriate?
No
Is the presentation of the work clear?
Yes
Are the images in this manuscript (including electrophoretic gels and blots) free
from apparent manipulation?
Yes
Please review this paper.
user
Reviewer’s report
Title: 
May chronic cough in chronic obstructive pulmonary disease be a
contraindication of Percutaneous Endoscopic Gastrostomy placement: a case
report
Version: 
5 
Date: 
15 Oct 2020
Reviewer's report:
PEER REVIEWER ASSESSMENTS: 
OBJECTIVE - Full research articles: is there a clear objective that addresses one
or several testable research questions? (Brief or other article types: is there a
clear objective?)
Yes - there is a clear objective
DESIGN - Is the current approach (including controls and analysis protocols)
appropriate for the objective?
Yes - the approach is appropriate
EXECUTION - Are the experiments and analyses performed with sufficient
technical rigor to allow confidence in the results?
No - there are minor issues
STATISTICS - Is the use of statistics in the manuscript appropriate?
N/A - there are no statistics in this study
INTERPRETATION - Is the current interpretation/discussion of the results
reasonable and not overstated?
No - there are minor issues
OVERALL MANUSCRIPT POTENTIAL - Has the author addressed your
concerns sufficiently for you to now recommend the work as a technically sound
contribution? If not, can further revisions be made to make the work technically
sound?
Probably - with minor revisions
PEER REVIEWER COMMENTS: 
GENERAL COMMENTS: Please note I have done this review as a "re-review"
and have highlighted the points of a re-review; 
I did not do the first review and
this evaluation will still address the points that are important. 
HAVE THE AUTHORS RESPONDED ADEQUATELY TO EACH OF THE
QUESTIONS YOU RAISED IN YOUR COMMENTS 
AND WHAT HAVE
THE AUTHORS DONE WELL
The authors have done an excellent job responding to the comprehensive past 3
reviews! 
Well done authors!
HAVE THEY MADE THE APPROPRIATE CHANGES IN THE
MANUSCRIPT 
The authors have made valuable changes that have increased the knowledge of
the reader with VERY useful additional discussion
WHAT IS YOUR OVERALL IMPRESSION OF THE REVISED
MANUSCRIPT 
There is a definite message here but quite frankly I think the message is two-fold.
1. Do not pull the PEG tube up against the abdominal wall too tightly
2. Appropriate tube care is essential
IN WHAT WAYS DOES IT NOT MEET BEST PRACTICE
This revision DOES MEET BEST PRACTOICE
REQUESTED REVISIONS: 
ARE THERE ANY ADDITIONAL REVISIONS THE AUTHORS SHOULD
MAKE
Let me start by saying that I am not one of the original reviewers thus some of
my comments will be new to the authors
1. Abstract line 13 change " dislodgement" to " erosion" 
2. Abstract 
Lines 18-19 reword to "We report the development of BBS in a man
with chronic obstructive pulmonary disease who had 
a persistent chronic
morning cough and required placement of a PEG tube for continuous infusion of
Levodopa/carbidopa intestinal gel 
for advanced Parkinson's disease. 
3 abstract line 25 
add the words " an absolute " 
after the word " representing" 
4. Abstract line 27 add the phrase "regimen of " after the word "appropriate "
5. In the future please number the pages or use a continuous numbering of lines.
All further suggestions will be according to pages with the abstract as page 1 ( as
a reviewer I do not see the title page)
6. page 2 line 14 add the words " and enteral access" after the word nutrition". In
addition 
start the next sentence with "In addition, the PEG tube 
can be used to
intubate the proximal jejunum with an extension tube (J-PEG) to allow the
pharmacological treatment *****"
7. I would not use the abbreviation ADT; it is not a common one
8. Page 2 line 21 start a new paragraph
9. Page 2 line 26. Which part of the bowel? I think you mean "colon during
insertion"
10. Page 2 line 30 change "migrates" to "erodes"
11. Page 2 line 36 what do you mean by leakage of fluid "in the peristomal site"?
Don't you mean "leakage of gastric content from around the PEG tube insertion
site or into the peritoneum"
12. Page 2 line 36 what do you mean about immobility of the PEG tube- You
need to be more clear. Similarly on line 37 " inability to insert. WHAT? The
tube?, fluid? 
13. Page 2 line 52 change to "consent"
14. Page 3 line10 change "routinely" to "routine" and delete 
"control" and
replace by "post insertion endoscopic confirmation of satisfactory PEG
placement"
15 I am not impressed by figures 1 and 2 moreover they should be figure 1A and
B; but I would 
delete them
16. Page 3 line 21 change the word "stoma" to "site" 
And add at the end " and "with visualization of only the J-tube extension" 
17. Page 3 line 10 here is where you should define the type of PEG tube you
used and again on line 30. 
After which delete lines 36-37
18. Page 3 lines 32-35. How does this second situation differ from the first one ?
They seem similar so why not try a needle knife-assisted approach? If the
findings are different please explain better
19. Page 3 lines 48-49. I will have to disagree that the morning cough was not
responsible 
rather the tube was positioned with too much tension placed on the
bumpers and the real problem was the lack of appropriate tube management "
sorry 
if the patient had a persistent cough 
all the time then I might relate it to
coughing, but you say that this was only a morning cough. Therefore I 
cannot
believe it was the cough that did this
20. Page 3 lines 57-59 I do not think this discussion is appropriate. There was too
much traction on the tube and may be that the coughing exacerbated it, but it was
not the coughing that caused this
21. Page 3 line 57. The word "decubitus" is not appropriate 
22. Page 4 lines 6-10 you need to define these types not everyone will have
ready access to the 
internet to review these papers
23. Page 4 lines 13 why are pharyngeal and /or esophageal obstruction a
contraindication? 
That makes no sense to me -indeed obstruction on these
regions is why a PEG may be needed 
24. Page 4 lines 26-28 I do not see how either a radiologic or operative approach
will prevent the care taker of pulling the tube up to the abdominal wall too
tightly. You need to describe how an operative approach will prevent this
25. Page 4 line 32 add to the end of the sentence "as well as to ability to feed
directly into the jejunum"
26. Page 4 line 38 change into and outside of the stomach" to "advanced further
in to the stomach and them pulled back until set to the correct length and
tension"
27 page 4 line 40 change to "the tube needs to have some slack to
accommodate***
28. Page 4 lines 47-49. This explanation makes no sense - the J tube does not
erode the balloon
29. Page 4 line 53 again change migration to erosion 
30. Sorry for all the suggestions, but fixing these will make the report much
clearer
ADDITIONAL REQUESTS/SUGGESTIONS: 
see bove
Publons Reviewer Recognition
. Springer Nature can send verification of this
review directly to Publons (a subsidiary of Clarivate Analytics). If you would
like to take advantage of this service, please click on the “Yes” option below.
Your name, email address, title of the reviewed manuscript, name of the journal,
and date of your review submission (the “Review Data”) will then be transmitted
to Publons upon publication of the manuscript. If you have already registered at
Publons, they will notify you of the receipt of this review and update your profile
as per your settings and their policy. If you are not registered with Publons, you
will receive an email from them asking you to register in order for them to be
able to recognize your review on your new profile page. Publons may use the
Review Data to generate derivative metadata for the benefit of Publons and you
as a reviewer, carefully considering the sensitivity of such information. For
example, Publons may verify your record as a reviewer by updating your profile
published on its webservice if you have registered for such service or help
editors to identify candidate reviewers. Please find the details of processing in
Publons’ privacy policy 
https://publons.com/about/terms
No
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 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.
This reviewer has been recruited by a partner organization, Research Square.
Reviewers with declared or apparent competing interests are not utilized for
these reviews. This reviewer has agreed to publication of their comments online
under a Creative Commons Attribution License attributed to Research Square
and was paid a small honorarium for completing the review within a specified
timeframe. Honoraria for reviews such as this are paid regardless of the reviewer
recommendation.
Reviewer Publication Consent.
 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
) if this manuscript is accepted for
publication. Any comments that I do not wish to be included in the published
report have been included as confidential comments to the editor, which will not
be published.
If you are not happy for us to publish this report, please contact the editorial
office before completing the review.
If you wish, you can include your name in your published report. Please note
you must decide whether to include your name at the start of the process and
confirm this decision whenever prompted. If you change your decision later, we
will not be able to publish your name.
I agree to the terms of the CC-BY 4.0 license; please publish my name with my
report.
Is the study design appropriate to answer the research question (including the use
of appropriate controls), and are the conclusions supported by the evidence
presented?
Yes
Are the methods sufficiently described to allow the study to be repeated?
Yes
Is the use of statistics and treatment of uncertainties appropriate?
No
Is the presentation of the work clear?
Yes
Are the images in this manuscript (including electrophoretic gels and blots) free
from apparent manipulation?
Yes
assistant