Reviewer's report
Title:High mobility group B1 mediates gut bacterial translocation in
acetaminophen hepatotoxicity
Version: 1Date:4 June 2013
Reviewer: Carol R Gardner
Reviewer's report:
The authors addressed many of the issues that were brought up in the prior
review of the manuscript, however, there are still some points which need to be
addressed. Tenses and plurals need to be checked as there are sentences such
as “….and this Invitrogen PCNA kits is ….”. Many sections of text, particularly in
the results section, contain long, run-on sentences which are very difficult to
understand.
Major compulsory revisions:
In the methods and materials, what is the route of anti-HMGB1 administration?
From what route was blood collected for the ALT measurements? Please define
KHBB.
In the results section discussing hepatic mRNA expression, the authors state that
sham IgG treatment increased mRNA levels of TNF and IL-6. Was this compared
to saline only control or APAP alone? These data should have been compared to
APAP alone as that is what is causing the up-regulation of TNF and IL-6 at 24 h,
not the sham IgG.
The authors provided micrographs to support their contention that the cells
labeled with PCNA were nucleated cells. Unfortunately, these micrographs
demonstrated a lack of specificity of the PCNA antibody in their tissue or a
cross-reaction with the Invitrogen anti-HMGB1 antibody circulating in the blood
stream. The only labeled cells in the liver sections are erythrocytes and, in the
blood vessel, both erythrocytes and neutrophils appear to be labeled. If the
PCNA data are an important part of this manuscript, then the authors should try a
PCNA antibody from another vendor. The BrdU data are much more convincing
and enlarged inserts should be included. In addition, careful higher power
analysis of the BrdU images may indicate some positive non-parenchymal cells
as well. Thus, the PCNA data should be deleted from the manuscript. In addition,
the authors referenced a paper by Vaquero to support their PCNA data, which
supposedly enumerated proliferating non-parenchymal cells. Vaquero analyzed
only proliferating hepatocytes using a BrdU method.
The reporting of the hepatic BrdU data states that the number of BrdU-labeled
nuclei was significantly increased in both sham IgG and anti-HMGB1 groups. To
what are the authors comparing these data?
In the discussion, the authors suggest that inflammatory macrophages, along
with hepatocytes, may be responsible for the increase in TNF and IL-6 48 h
post-acetaminophen. While macrophages would be increased by 24 h,
regeneration is clearly occurring by 48 h, thus the macrophages might not be
inflammatory but anti-inflammatory, or a combination of the different populations.
The added paragraph on the correlation of gut permeability and bacterial
translocation is poorly written and needs to be edited.
Minor essential revisions:
Please rewrite the sections on serum HMGB1 concentrations and bacterial
translocation to MLN and the liver, the phrasing is very awkward and difficult to
understand.
The backgrounds of the micrographs in fig. 4 do not match.
There is no magnification stated for the main micrographs in figures 4 or 6,
although the authors have included a magnification for the figure 6C insets. If
PCNA data are to be included in another revised manuscript, insets should be
included for figure 6D as well.
Level of interest: An article whose findings are important to those with closely
related research interests
Quality of written English: Needs some language corrections before being
published
Statistical review: No, the manuscript does not need to be seen by a
statistician.
Declaration of competing interests:
I declare that I have no competing interests
Reviewer's report
Title:High mobility group B1 mediates gut bacterial translocation in
acetaminophen hepatotoxicity
Version: 1Date:4 June 2013
Reviewer: Carol R Gardner
Reviewer's report:
The authors addressed many of the issues that were brought up in the prior
review of the manuscript, however, there are still some points which need to be
addressed. Tenses and plurals need to be checked as there are sentences such
as “….and this Invitrogen PCNA kits is ….”. Many sections of text, particularly in
the results section, contain long, run-on sentences which are very difficult to
understand.
Major compulsory revisions:
In the methods and materials, what is the route of anti-HMGB1 administration?
From what route was blood collected for the ALT measurements? Please define
KHBB.
In the results section discussing hepatic mRNA expression, the authors state that
sham IgG treatment increased mRNA levels of TNF and IL-6. Was this compared
to saline only control or APAP alone? These data should have been compared to
APAP alone as that is what is causing the up-regulation of TNF and IL-6 at 24 h,
not the sham IgG.
The authors provided micrographs to support their contention that the cells
labeled with PCNA were nucleated cells. Unfortunately, these micrographs
demonstrated a lack of specificity of the PCNA antibody in their tissue or a
cross-reaction with the Invitrogen anti-HMGB1 antibody circulating in the blood
stream. The only labeled cells in the liver sections are erythrocytes and, in the
blood vessel, both erythrocytes and neutrophils appear to be labeled. If the
PCNA data are an important part of this manuscript, then the authors should try a
PCNA antibody from another vendor. The BrdU data are much more convincing
and enlarged inserts should be included. In addition, careful higher power
analysis of the BrdU images may indicate some positive non-parenchymal cells
as well. Thus, the PCNA data should be deleted from the manuscript. In addition,
the authors referenced a paper by Vaquero to support their PCNA data, which
supposedly enumerated proliferating non-parenchymal cells. Vaquero analyzed
only proliferating hepatocytes using a BrdU method.
The reporting of the hepatic BrdU data states that the number of BrdU-labeled
nuclei was significantly increased in both sham IgG and anti-HMGB1 groups. To
what are the authors comparing these data?
In the discussion, the authors suggest that inflammatory macrophages, along
with hepatocytes, may be responsible for the increase in TNF and IL-6 48 h
post-acetaminophen. While macrophages would be increased by 24 h,
regeneration is clearly occurring by 48 h, thus the macrophages might not be
inflammatory but anti-inflammatory, or a combination of the different populations.
The added paragraph on the correlation of gut permeability and bacterial
translocation is poorly written and needs to be edited.
Minor essential revisions:
Please rewrite the sections on serum HMGB1 concentrations and bacterial
translocation to MLN and the liver, the phrasing is very awkward and difficult to
understand.
The backgrounds of the micrographs in fig. 4 do not match.
There is no magnification stated for the main micrographs in figures 4 or 6,
although the authors have included a magnification for the figure 6C insets. If
PCNA data are to be included in another revised manuscript, insets should be
included for figure 6D as well.
Level of interest: An article whose findings are important to those with closely
related research interests
Quality of written English: Needs some language corrections before being
published
Statistical review: No, the manuscript does not need to be seen by a
statistician.
Declaration of competing interests:
I declare that I have no competing interests
Please review this paper.
user
Reviewer's report
Title:High mobility group B1 mediates gut bacterial translocation in
acetaminophen hepatotoxicity
Version: 1Date:4 June 2013
Reviewer: Carol R Gardner
Reviewer's report:
The authors addressed many of the issues that were brought up in the prior
review of the manuscript, however, there are still some points which need to be
addressed. Tenses and plurals need to be checked as there are sentences such
as “….and this Invitrogen PCNA kits is ….”. Many sections of text, particularly in
the results section, contain long, run-on sentences which are very difficult to
understand.
Major compulsory revisions:
In the methods and materials, what is the route of anti-HMGB1 administration?
From what route was blood collected for the ALT measurements? Please define
KHBB.
In the results section discussing hepatic mRNA expression, the authors state that
sham IgG treatment increased mRNA levels of TNF and IL-6. Was this compared
to saline only control or APAP alone? These data should have been compared to
APAP alone as that is what is causing the up-regulation of TNF and IL-6 at 24 h,
not the sham IgG.
The authors provided micrographs to support their contention that the cells
labeled with PCNA were nucleated cells. Unfortunately, these micrographs
demonstrated a lack of specificity of the PCNA antibody in their tissue or a
cross-reaction with the Invitrogen anti-HMGB1 antibody circulating in the blood
stream. The only labeled cells in the liver sections are erythrocytes and, in the
blood vessel, both erythrocytes and neutrophils appear to be labeled. If the
PCNA data are an important part of this manuscript, then the authors should try a
PCNA antibody from another vendor. The BrdU data are much more convincing
and enlarged inserts should be included. In addition, careful higher power
analysis of the BrdU images may indicate some positive non-parenchymal cells
as well. Thus, the PCNA data should be deleted from the manuscript. In addition,
the authors referenced a paper by Vaquero to support their PCNA data, which
supposedly enumerated proliferating non-parenchymal cells. Vaquero analyzed
only proliferating hepatocytes using a BrdU method.
The reporting of the hepatic BrdU data states that the number of BrdU-labeled
nuclei was significantly increased in both sham IgG and anti-HMGB1 groups. To
what are the authors comparing these data?
In the discussion, the authors suggest that inflammatory macrophages, along
with hepatocytes, may be responsible for the increase in TNF and IL-6 48 h
post-acetaminophen. While macrophages would be increased by 24 h,
regeneration is clearly occurring by 48 h, thus the macrophages might not be
inflammatory but anti-inflammatory, or a combination of the different populations.
The added paragraph on the correlation of gut permeability and bacterial
translocation is poorly written and needs to be edited.
Minor essential revisions:
Please rewrite the sections on serum HMGB1 concentrations and bacterial
translocation to MLN and the liver, the phrasing is very awkward and difficult to
understand.
The backgrounds of the micrographs in fig. 4 do not match.
There is no magnification stated for the main micrographs in figures 4 or 6,
although the authors have included a magnification for the figure 6C insets. If
PCNA data are to be included in another revised manuscript, insets should be
included for figure 6D as well.
Level of interest: An article whose findings are important to those with closely
related research interests
Quality of written English: Needs some language corrections before being
published
Statistical review: No, the manuscript does not need to be seen by a
statistician.
Declaration of competing interests:
I declare that I have no competing interests
assistant