Understanding the Pattern of Oropharyngeal Cancers from North-East Romanian Patients

Round 1

Reviewer 1 Report

In this article, Ursu and colleagues reported the results of a study conducted on formalin fixed paraffin embedded (FFPE) collected from Romanian patients to assess the role of HPV in selected oropharyngeal squamous cellular cancers (OPSCC) using molecular assay.

Authors reported the results obtained from 54 samples tested to detect the presence of HPV DNA and RNA, p16 expression and FGFR3 mutation status.

 

It is important to well define the course of Human Papillomavirus (HPV) infection and its association with the development of OPSCC to improve diagnosis and prevention programs. Even if a study of this type must be predominantly descriptive, data reported are interesting and could increase the results already reported in literature.

However, results obtained are very limited and approximately described. Methods used should be descried more in details or some reference should be added to better explained techniques used to analyze FFPE samples. For example, no description regarding nucleic acids extraction is reported.

The use of English language should be revised in some parts.

 

There are major revisions that require attention by the authors.

 

Introduction:

• Authors should better describe why is important to study FGFR3 mutation status in patients with OPSCC
• Line 85: Has OPC the same meaning of OPSCC?
• Line 90: Authors should report the meaning of ASR

 

Materials and Methods:

• Line 106: Authors should better describe the number of types of OPSCC samples analyzed.
• Lines 122-123: Authors should insert reference regarding HPV DNA and RNA detection.
• A description of nucleic acids extraction method used should be added

 

Results:

• No statistical analysis is present.
• Table 1: Authors should specify NOS.

 

Discussion:

The discussion section is divided in paragraph as a review paper, but this manuscript is reports as a research article. Different studies reported in the Discussion section as reference articles reported a higher number of tested samples compared to that reported in the Results. Authors should better describe the limits of this study.

 

Conclusions:

• Line 322: Authors reported: “We have detected a higher concordance between HPV DNA and p16 than between HPV DNA and RNA”. Is this difference statistically significant?

Author Response

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Author Response File: Author Response.docx

Reviewer 2 Report

In the manuscript entitled: “Understanding the pattern of oropharyngeal cancers from North-East Romanian patients”, Ramona Gabriela Ursu and colleagues assessed the role of HPV in selected OPSCC cases, from Romanian patients by sensitive and complementary molecular assay.

The presentation and rationale of the study are not accurately described, several errors were found in the text that should be extensively reviewed throughout the paper, from the introduction, to the methods and in the discussion.

I list just a few of the many factors that are not appropriate:

1- Line 58-59 Recheck the logical sense of the sentence

2- Insert the definition at the first appearance of the acronym

3- Line 73-74 what is the message? What type of therapeutic approach is involved?

4- The introduction is not adequately focused on the goals of the experimental work.

5- The structure of the work does not make possible the reproducibility of the data exposed, there are not the defined sections of an experimental work: Materials and methods must be detailed, the exact number of patients analyzed is not reported

6- The functions of FGFR3 mutation status is not introduced

7- In tab-1_ recheck the tab title should be generic for HPV, since all clinical data of the 18 patients positive for both HPV16 and HPV18 should be reported.

It is suggested that the authors should focus their work better, choosing to do experimental article or literature review  article.

Author Response

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Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

After revisions, authors have responded to the requests made and expanded the manuscript.

Authors introduced a new paragraph in the Introduction section describing FGFR3 mutation status as potential biomarker in patients with OPSCC. Discussion has been improved and limits of this study has been better described.

However, Materials and Methods section is still not described in details. Authors have added he nucleic acids extraction kit used but they should also reported which is the starting amount of formalin fixed paraffin embedded samples used for nucleic acids extraction.

Moreover, authors should describe in detail the protocols used for biomarker analysis. For example, volume of reagents for amplification analysis, thermal profiles used, the volume of extracted DNA or RNA for each reaction… This should help the reproducibility of results obtained and the reader to better understand specific experimental procedures used.

Results even if limited should be divided into paragraphs based on the Materials and Methods subdivisions.

Some minor revisions are still necessary to improve the manuscript.

Author Response

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Author Response File: Author Response.docx

Reviewer 2 Report

The paper has been improved from the first version, but unfortunately it is not yet suitable for publication. It needs to be carefully rechecked by the authors so that anyone reading the paper can have a clear opinion about what the research has shown.

Beyond that it is also suggested:

To detail the method "Extraction of nucleic acids from FFP", or report a reference that has already used it.

At the beginning of the results specify the subject: 
Line 170-171 "18/54 (33.4%) were DNA/HPV positive..."
tissue samples? or patients?

It is suggested that the numbers and characteristics of all patients and tissues analyzed in this study be reported in Table 1.

The specimen number codes shown in Figure 1, "The p16-positive oropharyngeal tumors." to which patients correspond from Table 1? It is suggested that this be written in the caption of Figure 1.

Author Response

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Author Response File: Author Response.docx