Reviewer ’s report  
 
Title: Mitotic and apoptotic activity in colorectal neoplasia  
 
Version: 0 Date: 03 Nov 2017  
 
Reviewer: Michael Bordonaro  
 
Reviewer's report:  
 
This is an interesting contribution to the field and one which I, as a colorectal cancer researcher, 
found u seful.  Therefore I recommend acceptance, although with some minor revision:  
1. There are some spelling and grammar errors in the paper; these should be corrected.  
 
2. The authors correctly point out small sample sizes as a limitation of their study. Ano ther 
limitation worth noting is that most of the samples taken from healthy patients were from the 
rectum.  Now, I understand that no differences were found between neoplastic epithelial cells 
from different parts of the colon and rectum in patient with ab normality (question: since 
ostensibly normal tissue samples were also taken, 10 cm from the neoplasms, in affected 
patients, does this mean that these normal tissues were not significantly different in different 
parts of the colon and rectum in these patie nts?).  However, considering healthy patients with 
no neoplasm, we need to remember that different parts of the colon/rectum vary both 
physiologically as well as with respect to gene expression, so rectal samples in normal 
individuals are not necessarily r epresentative of the entire colorectum.  
 
3. The Discussion was somewhat disjointed and repetitious.  It would be better, for example, to 
consolidate all the paragraphs giving a background about apoptosis in one section; this would 
allow the Discussion to f low better and may allow the removal of superfluous sentences.  
 
4. The authors claim that the decrease in apoptosis observed with the transformation into 
carcinomas somehow contradicts the assumptions of the Warburg effect, presumably n 
reference to butyra te and apoptosis.  It would seem possible that this decrease in apoptosis 
results from changes in colonic cell physiology and gene expression, associated with 
neoplastic progression, that enable the cells to become more resistant to apoptotic signals.  
How ever, it does not logically follow that these cells are not experiencing increased levels of 
butyrate and that the butyrate is not inducing apoptosis.  Overall apoptosis may be lower with 
carcinoma, but it is still possible that carcinoma + high butyrate ( more dietary fiber?) would 
yield greater apoptosis than carcinoma + low butyrate.  Even more importantly, there has 
been discussion in the literature that butyrate is likely most effective as an anti -cancer factor 
at the early stages of colonic neoplasia ( early adenoma).  Thus, LT97 microadenoma cells are 
very sensitive to the effects of butyrate on apoptosis and proliferation suppression, more so 
than carcinoma cells.  The authors note that adenoma has higher apoptosis than carcinoma, 
and indeed adenoma wa s higher with respect to apoptosis than normal tissue.  Thus, adenoma 
cells may be particularly primed to be responsive to butyrate, a point that should be made in 
the Discussion.  
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.  
 
Yes 
 
Are you able to assess any statistics in t he 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 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 share s 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 rece ived 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 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 for publication, my named report  
including any attachments I upload will be posted on the websi te 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 name d 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: Mitotic and apoptotic activity in colorectal neoplasia  
 
Version: 0 Date: 03 Nov 2017  
 
Reviewer: Michael Bordonaro  
 
Reviewer's report:  
 
This is an interesting contribution to the field and one which I, as a colorectal cancer researcher, 
found u seful.  Therefore I recommend acceptance, although with some minor revision:  
1. There are some spelling and grammar errors in the paper; these should be corrected.  
 
2. The authors correctly point out small sample sizes as a limitation of their study. Ano ther 
limitation worth noting is that most of the samples taken from healthy patients were from the 
rectum.  Now, I understand that no differences were found between neoplastic epithelial cells 
from different parts of the colon and rectum in patient with ab normality (question: since 
ostensibly normal tissue samples were also taken, 10 cm from the neoplasms, in affected 
patients, does this mean that these normal tissues were not significantly different in different 
parts of the colon and rectum in these patie nts?).  However, considering healthy patients with 
no neoplasm, we need to remember that different parts of the colon/rectum vary both 
physiologically as well as with respect to gene expression, so rectal samples in normal 
individuals are not necessarily r epresentative of the entire colorectum.  
 
3. The Discussion was somewhat disjointed and repetitious.  It would be better, for example, to 
consolidate all the paragraphs giving a background about apoptosis in one section; this would 
allow the Discussion to f low better and may allow the removal of superfluous sentences.  
 
4. The authors claim that the decrease in apoptosis observed with the transformation into 
carcinomas somehow contradicts the assumptions of the Warburg effect, presumably n 
reference to butyra te and apoptosis.  It would seem possible that this decrease in apoptosis 
results from changes in colonic cell physiology and gene expression, associated with 
neoplastic progression, that enable the cells to become more resistant to apoptotic signals.  
How ever, it does not logically follow that these cells are not experiencing increased levels of 
butyrate and that the butyrate is not inducing apoptosis.  Overall apoptosis may be lower with 
carcinoma, but it is still possible that carcinoma + high butyrate ( more dietary fiber?) would 
yield greater apoptosis than carcinoma + low butyrate.  Even more importantly, there has 
been discussion in the literature that butyrate is likely most effective as an anti -cancer factor 
at the early stages of colonic neoplasia ( early adenoma).  Thus, LT97 microadenoma cells are 
very sensitive to the effects of butyrate on apoptosis and proliferation suppression, more so 
than carcinoma cells.  The authors note that adenoma has higher apoptosis than carcinoma, 
and indeed adenoma wa s higher with respect to apoptosis than normal tissue.  Thus, adenoma 
cells may be particularly primed to be responsive to butyrate, a point that should be made in 
the Discussion.  
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.  
 
Yes 
 
Are you able to assess any statistics in t he 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 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 share s 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 rece ived 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 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 for publication, my named report  
including any attachments I upload will be posted on the websi te 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 name d 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: Mitotic and apoptotic activity in colorectal neoplasia  
 
Version: 0 Date: 03 Nov 2017  
 
Reviewer: Michael Bordonaro  
 
Reviewer's report:  
 
This is an interesting contribution to the field and one which I, as a colorectal cancer researcher, 
found u seful.  Therefore I recommend acceptance, although with some minor revision:  
1. There are some spelling and grammar errors in the paper; these should be corrected.  
 
2. The authors correctly point out small sample sizes as a limitation of their study. Ano ther 
limitation worth noting is that most of the samples taken from healthy patients were from the 
rectum.  Now, I understand that no differences were found between neoplastic epithelial cells 
from different parts of the colon and rectum in patient with ab normality (question: since 
ostensibly normal tissue samples were also taken, 10 cm from the neoplasms, in affected 
patients, does this mean that these normal tissues were not significantly different in different 
parts of the colon and rectum in these patie nts?).  However, considering healthy patients with 
no neoplasm, we need to remember that different parts of the colon/rectum vary both 
physiologically as well as with respect to gene expression, so rectal samples in normal 
individuals are not necessarily r epresentative of the entire colorectum.  
 
3. The Discussion was somewhat disjointed and repetitious.  It would be better, for example, to 
consolidate all the paragraphs giving a background about apoptosis in one section; this would 
allow the Discussion to f low better and may allow the removal of superfluous sentences.  
 
4. The authors claim that the decrease in apoptosis observed with the transformation into 
carcinomas somehow contradicts the assumptions of the Warburg effect, presumably n 
reference to butyra te and apoptosis.  It would seem possible that this decrease in apoptosis 
results from changes in colonic cell physiology and gene expression, associated with 
neoplastic progression, that enable the cells to become more resistant to apoptotic signals.  
How ever, it does not logically follow that these cells are not experiencing increased levels of 
butyrate and that the butyrate is not inducing apoptosis.  Overall apoptosis may be lower with 
carcinoma, but it is still possible that carcinoma + high butyrate ( more dietary fiber?) would 
yield greater apoptosis than carcinoma + low butyrate.  Even more importantly, there has 
been discussion in the literature that butyrate is likely most effective as an anti -cancer factor 
at the early stages of colonic neoplasia ( early adenoma).  Thus, LT97 microadenoma cells are 
very sensitive to the effects of butyrate on apoptosis and proliferation suppression, more so 
than carcinoma cells.  The authors note that adenoma has higher apoptosis than carcinoma, 
and indeed adenoma wa s higher with respect to apoptosis than normal tissue.  Thus, adenoma 
cells may be particularly primed to be responsive to butyrate, a point that should be made in 
the Discussion.  
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.  
 
Yes 
 
Are you able to assess any statistics in t he 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 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 share s 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 rece ived 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 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 for publication, my named report  
including any attachments I upload will be posted on the websi te 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 name d 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