Reviewer ’s report  
 
Title: Mitotic and apoptotic  activity in colorectal neoplasia  
 
Version: 0 Date: 07 Dec 2017  
 
Reviewer: Anna Means  
 
Reviewer's report:  
 
This manuscript by Kohoutova et al. presents a descriptive study detailing the extent and pattern 
of proliferation and apoptosis as colonic epitheliu m progresses from normal to adenoma to 
carcinoma. They find that proliferation increases in early adenomas but that apoptosis also 
increases. In carcinomas, apoptosis is reduced while proliferation remains high. There is some 
variation in timing in differe nt regions of crypts (lower crypt, upper crypt, and surface cells) but 
the pattern is similar. Overall, the study is performed well with many variables accounted for 
(although much of this is "data not shown") and data are presented and analyzed well. The data 
are descriptive but interesting and support the dogma that tumorigenesis first involves increased 
proliferation then increased survival.  
 
Concerns:  
1. Since the main conclusion stated at the end of the manuscript is that proliferation and 
apoptosis a re similar between normal healthy donors and healthy adjacent tissue from cancer 
patients, those data should be shown as a graph, not just stated as "No differences… p<0.05" 
(page 9, line 1).  
 
2. Similarly, the authors state that for left and right sided co lon tissue "no differences were 
found in each pathology; p<0.05." The authors should show these data graphed so that the 
reader could know if there any increased variability in left vs. right sided colon. 
Additionally, if MSI status is available for these tumors, it would be helpful to know since 
these tumors (typically right sided) tend to have different characteristics that microsatellite 
stable tumors.  
 
3. Page 6, line 29: please clarify if it was 750 cells evaluated per patient sample as opposed to 
750 c ells totaled from all samples.  
 
4. Page 7, line 10: please clarify if it was 1500 cells evaluated per patient sample as opposed to 
1500 cells totaled from all samples.  
 
5. The Discussion section is overly long. The third and fourth paragraphs are not relevan t to 
the results and could be deleted.  
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 recommend additional statistical review  
 
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 r eceived 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, o r 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 ca n 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 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: Mitotic and apoptotic  activity in colorectal neoplasia  
 
Version: 0 Date: 07 Dec 2017  
 
Reviewer: Anna Means  
 
Reviewer's report:  
 
This manuscript by Kohoutova et al. presents a descriptive study detailing the extent and pattern 
of proliferation and apoptosis as colonic epitheliu m progresses from normal to adenoma to 
carcinoma. They find that proliferation increases in early adenomas but that apoptosis also 
increases. In carcinomas, apoptosis is reduced while proliferation remains high. There is some 
variation in timing in differe nt regions of crypts (lower crypt, upper crypt, and surface cells) but 
the pattern is similar. Overall, the study is performed well with many variables accounted for 
(although much of this is "data not shown") and data are presented and analyzed well. The data 
are descriptive but interesting and support the dogma that tumorigenesis first involves increased 
proliferation then increased survival.  
 
Concerns:  
1. Since the main conclusion stated at the end of the manuscript is that proliferation and 
apoptosis a re similar between normal healthy donors and healthy adjacent tissue from cancer 
patients, those data should be shown as a graph, not just stated as "No differences… p<0.05" 
(page 9, line 1).  
 
2. Similarly, the authors state that for left and right sided co lon tissue "no differences were 
found in each pathology; p<0.05." The authors should show these data graphed so that the 
reader could know if there any increased variability in left vs. right sided colon. 
Additionally, if MSI status is available for these tumors, it would be helpful to know since 
these tumors (typically right sided) tend to have different characteristics that microsatellite 
stable tumors.  
 
3. Page 6, line 29: please clarify if it was 750 cells evaluated per patient sample as opposed to 
750 c ells totaled from all samples.  
 
4. Page 7, line 10: please clarify if it was 1500 cells evaluated per patient sample as opposed to 
1500 cells totaled from all samples.  
 
5. The Discussion section is overly long. The third and fourth paragraphs are not relevan t to 
the results and could be deleted.  
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 recommend additional statistical review  
 
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 r eceived 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, o r 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 ca n 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 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: Mitotic and apoptotic  activity in colorectal neoplasia  
 
Version: 0 Date: 07 Dec 2017  
 
Reviewer: Anna Means  
 
Reviewer's report:  
 
This manuscript by Kohoutova et al. presents a descriptive study detailing the extent and pattern 
of proliferation and apoptosis as colonic epitheliu m progresses from normal to adenoma to 
carcinoma. They find that proliferation increases in early adenomas but that apoptosis also 
increases. In carcinomas, apoptosis is reduced while proliferation remains high. There is some 
variation in timing in differe nt regions of crypts (lower crypt, upper crypt, and surface cells) but 
the pattern is similar. Overall, the study is performed well with many variables accounted for 
(although much of this is "data not shown") and data are presented and analyzed well. The data 
are descriptive but interesting and support the dogma that tumorigenesis first involves increased 
proliferation then increased survival.  
 
Concerns:  
1. Since the main conclusion stated at the end of the manuscript is that proliferation and 
apoptosis a re similar between normal healthy donors and healthy adjacent tissue from cancer 
patients, those data should be shown as a graph, not just stated as "No differences… p<0.05" 
(page 9, line 1).  
 
2. Similarly, the authors state that for left and right sided co lon tissue "no differences were 
found in each pathology; p<0.05." The authors should show these data graphed so that the 
reader could know if there any increased variability in left vs. right sided colon. 
Additionally, if MSI status is available for these tumors, it would be helpful to know since 
these tumors (typically right sided) tend to have different characteristics that microsatellite 
stable tumors.  
 
3. Page 6, line 29: please clarify if it was 750 cells evaluated per patient sample as opposed to 
750 c ells totaled from all samples.  
 
4. Page 7, line 10: please clarify if it was 1500 cells evaluated per patient sample as opposed to 
1500 cells totaled from all samples.  
 
5. The Discussion section is overly long. The third and fourth paragraphs are not relevan t to 
the results and could be deleted.  
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 recommend additional statistical review  
 
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 r eceived 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, o r 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 ca n 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 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