Reviewer's report
Title:Neoplastic transformation of rat liver epithelial cells is enhanced by
non-transferrin-bound iron.
Version: 2Date:27 November 2007
Reviewer: Kyle E Brown
Reviewer's report:
General
The authors have provided a reasonable response to the critiques, although the
use of the MTT assay to address the issue of cytotoxicity is not wholly
satisfactory. The MTT assay measures a metabolic activity (reduction of a
tetrazolium compound) that is assumed to be proportional to the number of cells
present. In the current work, the MTT assay confirms the the overall result of the
cell counts but does not provide additional information regarding the mechanism
by which iron acts as a tumor promoter. This issue would be more appropriately
addressed by measurements of cell membrane integrity (e.g., Trypan Blue
exclusion, LDH release, etc.) or, better yet, reproductive integrity (clonogenic
survival). Since the effects of high-dose FAC on cell number were apparent as
early as 3 days, these parameters might be informative after a single exposure to
FAC.
-------------------------------------------------------------------------------
Major Compulsory Revisions (that the author must respond to before a decision
on publication can be reached)
-------------------------------------------------------------------------------
Minor Essential Revisions (such as missing labels on figures, or the wrong use of
a term, which the author can be trusted to correct)
On the top of page 16, the authors make reference to "local NTBI concentrations
that exist in necrotic regions of an iron loaded cirrhotic liver." While I accept the
point regarding potential differences in iron citrate levels between the liver and
the periphery, the implication that there is widespread necrosis in iron-loaded
liver (whether human disease or animal models) in incorrect and may be
misleading to readers who are not expert in this area. This should be changed.
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Discretionary Revisions (which the author can choose to ignore)
What next?: Accept after minor essential revisions
Level of interest: An article whose findings are important to those with closely
related research interests
Quality of written English: Acceptable
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:Neoplastic transformation of rat liver epithelial cells is enhanced by
non-transferrin-bound iron.
Version: 2Date:27 November 2007
Reviewer: Kyle E Brown
Reviewer's report:
General
The authors have provided a reasonable response to the critiques, although the
use of the MTT assay to address the issue of cytotoxicity is not wholly
satisfactory. The MTT assay measures a metabolic activity (reduction of a
tetrazolium compound) that is assumed to be proportional to the number of cells
present. In the current work, the MTT assay confirms the the overall result of the
cell counts but does not provide additional information regarding the mechanism
by which iron acts as a tumor promoter. This issue would be more appropriately
addressed by measurements of cell membrane integrity (e.g., Trypan Blue
exclusion, LDH release, etc.) or, better yet, reproductive integrity (clonogenic
survival). Since the effects of high-dose FAC on cell number were apparent as
early as 3 days, these parameters might be informative after a single exposure to
FAC.
-------------------------------------------------------------------------------
Major Compulsory Revisions (that the author must respond to before a decision
on publication can be reached)
-------------------------------------------------------------------------------
Minor Essential Revisions (such as missing labels on figures, or the wrong use of
a term, which the author can be trusted to correct)
On the top of page 16, the authors make reference to "local NTBI concentrations
that exist in necrotic regions of an iron loaded cirrhotic liver." While I accept the
point regarding potential differences in iron citrate levels between the liver and
the periphery, the implication that there is widespread necrosis in iron-loaded
liver (whether human disease or animal models) in incorrect and may be
misleading to readers who are not expert in this area. This should be changed.
-------------------------------------------------------------------------------
Discretionary Revisions (which the author can choose to ignore)
What next?: Accept after minor essential revisions
Level of interest: An article whose findings are important to those with closely
related research interests
Quality of written English: Acceptable
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:Neoplastic transformation of rat liver epithelial cells is enhanced by
non-transferrin-bound iron.
Version: 2Date:27 November 2007
Reviewer: Kyle E Brown
Reviewer's report:
General
The authors have provided a reasonable response to the critiques, although the
use of the MTT assay to address the issue of cytotoxicity is not wholly
satisfactory. The MTT assay measures a metabolic activity (reduction of a
tetrazolium compound) that is assumed to be proportional to the number of cells
present. In the current work, the MTT assay confirms the the overall result of the
cell counts but does not provide additional information regarding the mechanism
by which iron acts as a tumor promoter. This issue would be more appropriately
addressed by measurements of cell membrane integrity (e.g., Trypan Blue
exclusion, LDH release, etc.) or, better yet, reproductive integrity (clonogenic
survival). Since the effects of high-dose FAC on cell number were apparent as
early as 3 days, these parameters might be informative after a single exposure to
FAC.
-------------------------------------------------------------------------------
Major Compulsory Revisions (that the author must respond to before a decision
on publication can be reached)
-------------------------------------------------------------------------------
Minor Essential Revisions (such as missing labels on figures, or the wrong use of
a term, which the author can be trusted to correct)
On the top of page 16, the authors make reference to "local NTBI concentrations
that exist in necrotic regions of an iron loaded cirrhotic liver." While I accept the
point regarding potential differences in iron citrate levels between the liver and
the periphery, the implication that there is widespread necrosis in iron-loaded
liver (whether human disease or animal models) in incorrect and may be
misleading to readers who are not expert in this area. This should be changed.
-------------------------------------------------------------------------------
Discretionary Revisions (which the author can choose to ignore)
What next?: Accept after minor essential revisions
Level of interest: An article whose findings are important to those with closely
related research interests
Quality of written English: Acceptable
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