Reviewer's report
Title:Transporter function and cyclic AMP turnover in normal colonic mucosa
from patients with and without colorectal neoplasia
Version: 1Date:20 February 2012
Reviewer: Kacper Wojtal
Reviewer's report:
Major Compulsory Revisions:
1. In my opinion the fact that db-cAMP is a membrane permeable analog puts the
working hypothesis of transport of this molecule into question. Hence, the extent
of the passive uptake in the generation of SCC by this molecule must be
somehow considered and possibly verified. cAMP did not give rise to a strong
signal, possibly due to either lower influx rate as compared to db-cAMP or
hydrolysis by PDEs. These issues could be approached by using either another
cAMP analog or the experiment with cAMP should be performed in combination
with any of the phosphodiesterase inhibitors, such as IBMX.
2. The proposed involvement of PKA and PDEs (Discussion; page 8-9) in the
generation of SCC could provide the support for the observed differences
between control and neoplastic tissues. If not approached experimentally, this
aspect should be more elaborated in the discussion section of the manuscript.
Discretionary Revisions:
1. Despite determining mRNA levels and localization in the colonic tissues, one
would still consider checking the protein levels of implicated transporters in both
control and neoplastic specimens by Western blot.
2. Concerning the other transporters which could be possibly involved in the
uptake of cNTs: were ENTs and/or CNTs tested? Do they transport cAMP?
Minor Essential Revisions:
Microscopic images in figures 5 and 6 appear to have different optical settings,
which should be adjusted. Also, in my opinion instead of providing the info about
the objectives used in the figure legends, placing the appropriate size bars
directly on the images would be more informative.
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:Transporter function and cyclic AMP turnover in normal colonic mucosa
from patients with and without colorectal neoplasia
Version: 1Date:20 February 2012
Reviewer: Kacper Wojtal
Reviewer's report:
Major Compulsory Revisions:
1. In my opinion the fact that db-cAMP is a membrane permeable analog puts the
working hypothesis of transport of this molecule into question. Hence, the extent
of the passive uptake in the generation of SCC by this molecule must be
somehow considered and possibly verified. cAMP did not give rise to a strong
signal, possibly due to either lower influx rate as compared to db-cAMP or
hydrolysis by PDEs. These issues could be approached by using either another
cAMP analog or the experiment with cAMP should be performed in combination
with any of the phosphodiesterase inhibitors, such as IBMX.
2. The proposed involvement of PKA and PDEs (Discussion; page 8-9) in the
generation of SCC could provide the support for the observed differences
between control and neoplastic tissues. If not approached experimentally, this
aspect should be more elaborated in the discussion section of the manuscript.
Discretionary Revisions:
1. Despite determining mRNA levels and localization in the colonic tissues, one
would still consider checking the protein levels of implicated transporters in both
control and neoplastic specimens by Western blot.
2. Concerning the other transporters which could be possibly involved in the
uptake of cNTs: were ENTs and/or CNTs tested? Do they transport cAMP?
Minor Essential Revisions:
Microscopic images in figures 5 and 6 appear to have different optical settings,
which should be adjusted. Also, in my opinion instead of providing the info about
the objectives used in the figure legends, placing the appropriate size bars
directly on the images would be more informative.
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:Transporter function and cyclic AMP turnover in normal colonic mucosa
from patients with and without colorectal neoplasia
Version: 1Date:20 February 2012
Reviewer: Kacper Wojtal
Reviewer's report:
Major Compulsory Revisions:
1. In my opinion the fact that db-cAMP is a membrane permeable analog puts the
working hypothesis of transport of this molecule into question. Hence, the extent
of the passive uptake in the generation of SCC by this molecule must be
somehow considered and possibly verified. cAMP did not give rise to a strong
signal, possibly due to either lower influx rate as compared to db-cAMP or
hydrolysis by PDEs. These issues could be approached by using either another
cAMP analog or the experiment with cAMP should be performed in combination
with any of the phosphodiesterase inhibitors, such as IBMX.
2. The proposed involvement of PKA and PDEs (Discussion; page 8-9) in the
generation of SCC could provide the support for the observed differences
between control and neoplastic tissues. If not approached experimentally, this
aspect should be more elaborated in the discussion section of the manuscript.
Discretionary Revisions:
1. Despite determining mRNA levels and localization in the colonic tissues, one
would still consider checking the protein levels of implicated transporters in both
control and neoplastic specimens by Western blot.
2. Concerning the other transporters which could be possibly involved in the
uptake of cNTs: were ENTs and/or CNTs tested? Do they transport cAMP?
Minor Essential Revisions:
Microscopic images in figures 5 and 6 appear to have different optical settings,
which should be adjusted. Also, in my opinion instead of providing the info about
the objectives used in the figure legends, placing the appropriate size bars
directly on the images would be more informative.
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
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