Hsp60 Quantification in Human Gastric Mucosa Shows Differences between Pathologies with Various Degrees of Proliferation and Malignancy Grade

Round 1

Reviewer 1 Report

The peper is very interesting and very good structurate. I suggest few minor language and editing corrections.

Author Response

Response point-by-point to the Reviewer

Reviewer 1 comment (RC1): The paper is very interesting and very good structurate. I suggest few minor language and editing corrections.

Authors’ Reply (AR): We thank this Reviewer for this positive comment. Two authors are experts in English and have double-checked the text to ensure correct vocabulary and construction.

Author Response File: Author Response.pdf

Reviewer 2 Report

Pitruzzella et al present a study of HSP60 expression by IHC on a wide array of gastric pathologies. This topic is of interest and the results showing an increase of HSP60 expression from normal mucosa, inflammatory situation, pre-cancerous proliferation to highly agressive and proliferative cancerous cells are convincing. display of data from figures 1 could be improved :

- please provide the mean +/- stdev for each patient category within the text. The "average percentage" provided is not sufficient.

- in the figure 1 legend, please indicate again the number of patients per group.

Author Response

Response point-by-point to the Reviewer

 

Reviewer 2 comment (RC2): Pitruzzella et al present a study of HSP60 expression by IHC on a wide array of gastric pathologies. This topic is of interest and the results showing an increase of HSP60 expression from normal mucosa, inflammatory situation, pre-cancerous proliferation to highly agressive and proliferative cancerous cells are convincing. display of data from figures 1 could be improved:

Authors’ Reply (AR): We thank this Reviewer for this positive comment.

RC2.1: 1) please provide the mean +/- stdev for each patient category within the text. The "average percentage" provided is not sufficient.

AR: We have included the respective standard deviations in the text, as requested.

RC2.2: 2) in the figure 1 legend, please indicate again the number of patients per group.

AR: We have added the number of cases for each group in the figure legend, as requested.

Author Response File: Author Response.pdf

Reviewer 3 Report

Taking into consideration the reported increase of Hsp60 levels  in human neoplastic pathology of many organs, such as uterus, colon, prostate, thyroid and salivary gland and colon the authors have immunohistochemically analyzed different groups of gastric mucosa, either normal or inflamed as well as hyperplastic , metaplastic and neoplastic specimens.

The quantitative analysis has documented a progressive increase of average percentage of Hsp60 from normal mucosa (32.8) to mild/moderate gastritis (33.5), severe gastritis (51.8), hyperplastic polyps (58.4),   intestinal metaplasia (67.0), gastric dysplasia (89.4) and adenocarcinomas (92.5).

Although an evident difference between dysplasia and adenocarcinoma with the other gastric pathologies was appreciable, the diagnostic differential purposes for Hsp60 is quite debatable, due to the evident morphological differences. By contrast, the hypothesized mechanism as well as the Hsp60 role on gastric carcinogenesis may be shareable.

The paper may be suitable for the publication even if some improvements are required:

1) The title should be changed avoiding the comment about the diagnostic criterion;

2) The agreement between two histopathology should be reported as k statistics value;

3) Figures should be reconsidered utilizing low and high magnification paired fields, clearly showing the different percentages of Hsp60 staining reported in the results sections. All figures should be furnished at least 300 dpi.

4) A more accurate English grammar and style revision should be performed.

Author Response

Response point-by-point to the Reviewer

 

Reviewer 3 comment (RC3): Taking into consideration the reported increase of Hsp60 levels in human neoplastic pathology of many organs, such as uterus, colon, prostate, thyroid and salivary gland and colon the authors have immunohistochemically analyzed different groups of gastric mucosa, either normal or inflamed as well as hyperplastic, metaplastic and neoplastic specimens. The quantitative analysis has documented a progressive increase of average percentage of Hsp60 from normal mucosa (32.8) to mild/moderate gastritis (33.5), severe gastritis (51.8), hyperplastic polyps (58.4), intestinal metaplasia (67.0), gastric dysplasia (89.4) and adenocarcinomas (92.5).

Although an evident difference between dysplasia and adenocarcinoma with the other gastric pathologies was appreciable, the diagnostic differential purposes for Hsp60 is quite debatable, due to the evident morphological differences. By contrast, the hypothesized mechanism as well as the Hsp60 role on gastric carcinogenesis may be shareable. The paper may be suitable for the publication even if some improvements are required:

Authors’ Reply (AR): We thank the Reviewer for the helpful comments.

RC3.1: 1) The title should be changed avoiding the comment about the diagnostic criterion;

AR: We have changed the title of the article, following this Reviewer’s suggestion.

RC3.2: 2) The agreement between two histopathology should be reported as k statistics value;

AR: We have integrated into the text the K value for each group examined (NM, MMG, SG, HP, IM, GD and GC). On each of the 10 cases, the observers independently determined the positivity of the sample, subsequently calculated in percentage in a high-power field (HPF) at 400x of magnification and repeated for 10 HPF, as reported in the text. K values are now reported in the text flanking each subgroup examination.

 

RC3.3: 3) Figures should be reconsidered utilizing low and high magnification paired fields, clearly showing the different percentages of Hsp60 staining reported in the results sections. All figures should be furnished at least 300 dpi.

AR: In Figure 2 we used 200x and 400x magnification for all the groups to show the differences between them. In figures 3 and 4, we wanted to show some details that are commented on in the text. Figure 1 was provided at 600dpi, while the other figures were provided at 300dpi.

RC3.4: 4) A more accurate English grammar and style revision should be performed.

AR: One author is a native English speaker, and another specializes in languages, including English. Each has published numerous articles in English, in reputable scientific journals. They have also considerable experience as journal editors and referees. These authors have double-checked the manuscript to ensure language quality, following this Reviewer’s suggestion.

 

Author Response File: Author Response.pdf