Reviewer ’s report  
 
Title:  HPV positive, wild type TP53, and p16 overexpression correlate with the absence of 
residual tu mors after chemoradiotherapy in anal squamous cell carcinoma.  
 
Version: 0 Date: 04 Nov 2017 
 
Reviewer:  Veronika  Vymetalkova  
 
Reviewer’s report:  
 
Soares et al. aimed to investigate the impact of HPV and HIV status, p16 expression level and 
TP53 mutations as predictors of the local response i n Squamous Cell Carcinoma (SCC) of the 
anal canal.  
 
Although it is engaged in interesting topic, it has several important discrepancies which should 
be addressed. Also, the novelty of the study is limited. All above do together interact in a way 
that makes publication less relevant. I clearly suggest major revision.  
 
There are some typos and grammatical errors in the manuscript - therefore the final version would 
benefit from proofreading by native speaker or language editor.  
 
 
Introduction:  
 
The study lacks novelty, there are several other articles already published on all factors analyzed 
in this study. For this reason, the rationale for focusing on analyzed parameters should be more 
elaborated.  
 
Second paragraph in introduction is not clear, please reformulate it.  
 
When mention gene, use italics.  
 
 
Material methods:  
 
Why only 6 months follow up was analyzed, if patients have been recruited since 2003, it is 
beneficial if authors could analyze also overall and recurrence free interval. The majorit y of 
discussion is already based on prognostic factors of evaluated parameters.  
 
Why exon 1 of TP53 gene was not analysed for mutations?  
 
Page 7, row 24, the term "and" is redundant  
 
 
Results:  
Page 8, last three sentences in first paragraph in results sectio n could be merged together  
 
What about the concomitant analysis of all studied parameters? Are patients with any 
combination of parameters in higher risk for clinical response to chemoradiotherapy?  
 
Table 2 is difficult to understand, please define it more i n the result section. Especially, when 
mention the particular cases (cases 50,60, 19 and 10)  
 
 
Discussion:  
 
Discussion is very wordy and explaining the same results several times. This could be merged 
and thus shorten the text  
 
The statement about the importa nce of HPV to study for their role in SCC of the anal cancer is 
not very clear in the connection with head and neck cancer as stated in discussion, please enlarge 
this more.  
 
Authors state that homozygous DNA mutations can result from a loss of heterozygosi ty. The 
term homozygous refers to individuals that have identical alleles for a given trait. This part is not 
comprehensive. Please state clearly if it concerns allelic loss.  
 
Page 13, last paragraph, authors mentioned three mechanisms of explanation for c hromosome 17 
and tP53 mutations, however above are mention only two.  
 
Page 14, last paragraph is not clear at all, please reformulate it  
 
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.  
 
Unable to assess  
 
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 recomme nd 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 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.  
 
I declare that I have no competing interests.  
 
I agree to the open peer review policy of the journ al. 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 m y 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:  HPV positive, wild type TP53, and p16 overexpression correlate with the absence of 
residual tu mors after chemoradiotherapy in anal squamous cell carcinoma.  
 
Version: 0 Date: 04 Nov 2017 
 
Reviewer:  Veronika  Vymetalkova  
 
Reviewer’s report:  
 
Soares et al. aimed to investigate the impact of HPV and HIV status, p16 expression level and 
TP53 mutations as predictors of the local response i n Squamous Cell Carcinoma (SCC) of the 
anal canal.  
 
Although it is engaged in interesting topic, it has several important discrepancies which should 
be addressed. Also, the novelty of the study is limited. All above do together interact in a way 
that makes publication less relevant. I clearly suggest major revision.  
 
There are some typos and grammatical errors in the manuscript - therefore the final version would 
benefit from proofreading by native speaker or language editor.  
 
 
Introduction:  
 
The study lacks novelty, there are several other articles already published on all factors analyzed 
in this study. For this reason, the rationale for focusing on analyzed parameters should be more 
elaborated.  
 
Second paragraph in introduction is not clear, please reformulate it.  
 
When mention gene, use italics.  
 
 
Material methods:  
 
Why only 6 months follow up was analyzed, if patients have been recruited since 2003, it is 
beneficial if authors could analyze also overall and recurrence free interval. The majorit y of 
discussion is already based on prognostic factors of evaluated parameters.  
 
Why exon 1 of TP53 gene was not analysed for mutations?  
 
Page 7, row 24, the term "and" is redundant  
 
 
Results:  
Page 8, last three sentences in first paragraph in results sectio n could be merged together  
 
What about the concomitant analysis of all studied parameters? Are patients with any 
combination of parameters in higher risk for clinical response to chemoradiotherapy?  
 
Table 2 is difficult to understand, please define it more i n the result section. Especially, when 
mention the particular cases (cases 50,60, 19 and 10)  
 
 
Discussion:  
 
Discussion is very wordy and explaining the same results several times. This could be merged 
and thus shorten the text  
 
The statement about the importa nce of HPV to study for their role in SCC of the anal cancer is 
not very clear in the connection with head and neck cancer as stated in discussion, please enlarge 
this more.  
 
Authors state that homozygous DNA mutations can result from a loss of heterozygosi ty. The 
term homozygous refers to individuals that have identical alleles for a given trait. This part is not 
comprehensive. Please state clearly if it concerns allelic loss.  
 
Page 13, last paragraph, authors mentioned three mechanisms of explanation for c hromosome 17 
and tP53 mutations, however above are mention only two.  
 
Page 14, last paragraph is not clear at all, please reformulate it  
 
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.  
 
Unable to assess  
 
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 recomme nd 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 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.  
 
I declare that I have no competing interests.  
 
I agree to the open peer review policy of the journ al. 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 m y 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:  HPV positive, wild type TP53, and p16 overexpression correlate with the absence of 
residual tu mors after chemoradiotherapy in anal squamous cell carcinoma.  
 
Version: 0 Date: 04 Nov 2017 
 
Reviewer:  Veronika  Vymetalkova  
 
Reviewer’s report:  
 
Soares et al. aimed to investigate the impact of HPV and HIV status, p16 expression level and 
TP53 mutations as predictors of the local response i n Squamous Cell Carcinoma (SCC) of the 
anal canal.  
 
Although it is engaged in interesting topic, it has several important discrepancies which should 
be addressed. Also, the novelty of the study is limited. All above do together interact in a way 
that makes publication less relevant. I clearly suggest major revision.  
 
There are some typos and grammatical errors in the manuscript - therefore the final version would 
benefit from proofreading by native speaker or language editor.  
 
 
Introduction:  
 
The study lacks novelty, there are several other articles already published on all factors analyzed 
in this study. For this reason, the rationale for focusing on analyzed parameters should be more 
elaborated.  
 
Second paragraph in introduction is not clear, please reformulate it.  
 
When mention gene, use italics.  
 
 
Material methods:  
 
Why only 6 months follow up was analyzed, if patients have been recruited since 2003, it is 
beneficial if authors could analyze also overall and recurrence free interval. The majorit y of 
discussion is already based on prognostic factors of evaluated parameters.  
 
Why exon 1 of TP53 gene was not analysed for mutations?  
 
Page 7, row 24, the term "and" is redundant  
 
 
Results:  
Page 8, last three sentences in first paragraph in results sectio n could be merged together  
 
What about the concomitant analysis of all studied parameters? Are patients with any 
combination of parameters in higher risk for clinical response to chemoradiotherapy?  
 
Table 2 is difficult to understand, please define it more i n the result section. Especially, when 
mention the particular cases (cases 50,60, 19 and 10)  
 
 
Discussion:  
 
Discussion is very wordy and explaining the same results several times. This could be merged 
and thus shorten the text  
 
The statement about the importa nce of HPV to study for their role in SCC of the anal cancer is 
not very clear in the connection with head and neck cancer as stated in discussion, please enlarge 
this more.  
 
Authors state that homozygous DNA mutations can result from a loss of heterozygosi ty. The 
term homozygous refers to individuals that have identical alleles for a given trait. This part is not 
comprehensive. Please state clearly if it concerns allelic loss.  
 
Page 13, last paragraph, authors mentioned three mechanisms of explanation for c hromosome 17 
and tP53 mutations, however above are mention only two.  
 
Page 14, last paragraph is not clear at all, please reformulate it  
 
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.  
 
Unable to assess  
 
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 recomme nd 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 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.  
 
I declare that I have no competing interests.  
 
I agree to the open peer review policy of the journ al. 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 m y 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