Reviewer's report 
 Adenoviral Expression of a Transforming Growth Factor-beta1 Antisense mRNA is Effective in Title: 
Preventing Liver Fibrosis in Bile-Duct Ligated Rats 
 1  21 August 2003 Version: Date: 
 Serhiy Souchelnytskyi Reviewer: 
Reviewer's report: 
General 
The manuscript shows that expression of TGFbeta1 antisense mRNA resulted in decrease of liver fibrosis, as evaluated by a morphological analysis, deposition of collagen and expression of alpha-smooth muscle actin. 
The subject of study is of high importance and of interest for broad audience. 
This work is further development of studies of TGFbeta role in liver fibrosis. It presents new results consolidating previously published data, including those published by the authors. 
The manuscript is well written, results are presented clearly, and conclusions are supported by the 
data. 
Discretionary Revisions (which the author can choose to ignore) 
1. The authors convincingly showed high expression level of TGFbeta1 antisense mRNA. How is 
affected TGFbeta1 protein level? Do the authors have an estimation of how much active TGFbeta is 
still present in liver cells of Ad5-CMV-AS-TGFb1-infected animals? 
This question is because TGFbeta can be delivered from other organs, and/or it expression can be affected differently in various cells by Ad5-CMV-AS-TGFb1 infection. 
2. Did infection of Ad5-CMV-AS-TGFb1 have any systemic effect on rat? 
Previously it was published that an increased expression of TGFbeta1 can lead to loss of body 
weight and deterioration of a clinical status of animals. Thus, decrease of TGFbeta1 expression may 
have a systemic effect as well. 
On the other side, the short term of the experiments described in this manuscript may not be sufficient to see such changes, compared to reported longer observation terms. 
3. The authors use in the text expressions like ¿vector abrogates the TGF-beta-enchanced 
production¿¿ (page 5, as example), while there is no data showing that it is really TGFbeta-induced activity is affected. As I can see, it is shown that antisense mRNA of TGFbeta1 did block development of fibrosis. To state that the effect is on a TGFbeta-dependent activity, controlled 
TGFbeta-induced events have to be evaluated. 
4. Animal experiments were performed according to ¿strict governmental and international 
guidelines¿, which is a too general statement. Which guidelines were followed? 
5. Description of Figure 2, panels G and H, is very short. What do you mean as an obvious liver 
appearance? This is in contrast with the detailed description and presentation of a surgery procedure. 
6. Magnification bars in Figures 4 and 5 would be useful. 
Minor Compulsory Revisions (such as missing labels on figures, or the wrong use of a term, which 
the author can be trusted to correct) None 
Major Compulsory Revisions (that the author must respond to before a decision on publication can 
be reached) 
None 
 Accept after discretionary revisions Advice on publication: 
 A paper of considerable merit and interest in its field Level of interest: 
 Acceptable Quality of written English: 
Declaration of competing interests: 
None 
Reviewer's report 
 Adenoviral Expression of a Transforming Growth Factor-beta1 Antisense mRNA is Effective in Title: 
Preventing Liver Fibrosis in Bile-Duct Ligated Rats 
 1  21 August 2003 Version: Date: 
 Serhiy Souchelnytskyi Reviewer: 
Reviewer's report: 
General 
The manuscript shows that expression of TGFbeta1 antisense mRNA resulted in decrease of liver fibrosis, as evaluated by a morphological analysis, deposition of collagen and expression of alpha-smooth muscle actin. 
The subject of study is of high importance and of interest for broad audience. 
This work is further development of studies of TGFbeta role in liver fibrosis. It presents new results consolidating previously published data, including those published by the authors. 
The manuscript is well written, results are presented clearly, and conclusions are supported by the 
data. 
Discretionary Revisions (which the author can choose to ignore) 
1. The authors convincingly showed high expression level of TGFbeta1 antisense mRNA. How is 
affected TGFbeta1 protein level? Do the authors have an estimation of how much active TGFbeta is 
still present in liver cells of Ad5-CMV-AS-TGFb1-infected animals? 
This question is because TGFbeta can be delivered from other organs, and/or it expression can be affected differently in various cells by Ad5-CMV-AS-TGFb1 infection. 
2. Did infection of Ad5-CMV-AS-TGFb1 have any systemic effect on rat? 
Previously it was published that an increased expression of TGFbeta1 can lead to loss of body 
weight and deterioration of a clinical status of animals. Thus, decrease of TGFbeta1 expression may 
have a systemic effect as well. 
On the other side, the short term of the experiments described in this manuscript may not be sufficient to see such changes, compared to reported longer observation terms. 
3. The authors use in the text expressions like ¿vector abrogates the TGF-beta-enchanced 
production¿¿ (page 5, as example), while there is no data showing that it is really TGFbeta-induced activity is affected. As I can see, it is shown that antisense mRNA of TGFbeta1 did block development of fibrosis. To state that the effect is on a TGFbeta-dependent activity, controlled 
TGFbeta-induced events have to be evaluated. 
4. Animal experiments were performed according to ¿strict governmental and international 
guidelines¿, which is a too general statement. Which guidelines were followed? 
5. Description of Figure 2, panels G and H, is very short. What do you mean as an obvious liver 
appearance? This is in contrast with the detailed description and presentation of a surgery procedure. 
6. Magnification bars in Figures 4 and 5 would be useful. 
Minor Compulsory Revisions (such as missing labels on figures, or the wrong use of a term, which 
the author can be trusted to correct) None 
Major Compulsory Revisions (that the author must respond to before a decision on publication can 
be reached) 
None 
 Accept after discretionary revisions Advice on publication: 
 A paper of considerable merit and interest in its field Level of interest: 
 Acceptable Quality of written English: 
Declaration of competing interests: 
None 
Please review this paper.
user
Reviewer's report 
 Adenoviral Expression of a Transforming Growth Factor-beta1 Antisense mRNA is Effective in Title: 
Preventing Liver Fibrosis in Bile-Duct Ligated Rats 
 1  21 August 2003 Version: Date: 
 Serhiy Souchelnytskyi Reviewer: 
Reviewer's report: 
General 
The manuscript shows that expression of TGFbeta1 antisense mRNA resulted in decrease of liver fibrosis, as evaluated by a morphological analysis, deposition of collagen and expression of alpha-smooth muscle actin. 
The subject of study is of high importance and of interest for broad audience. 
This work is further development of studies of TGFbeta role in liver fibrosis. It presents new results consolidating previously published data, including those published by the authors. 
The manuscript is well written, results are presented clearly, and conclusions are supported by the 
data. 
Discretionary Revisions (which the author can choose to ignore) 
1. The authors convincingly showed high expression level of TGFbeta1 antisense mRNA. How is 
affected TGFbeta1 protein level? Do the authors have an estimation of how much active TGFbeta is 
still present in liver cells of Ad5-CMV-AS-TGFb1-infected animals? 
This question is because TGFbeta can be delivered from other organs, and/or it expression can be affected differently in various cells by Ad5-CMV-AS-TGFb1 infection. 
2. Did infection of Ad5-CMV-AS-TGFb1 have any systemic effect on rat? 
Previously it was published that an increased expression of TGFbeta1 can lead to loss of body 
weight and deterioration of a clinical status of animals. Thus, decrease of TGFbeta1 expression may 
have a systemic effect as well. 
On the other side, the short term of the experiments described in this manuscript may not be sufficient to see such changes, compared to reported longer observation terms. 
3. The authors use in the text expressions like ¿vector abrogates the TGF-beta-enchanced 
production¿¿ (page 5, as example), while there is no data showing that it is really TGFbeta-induced activity is affected. As I can see, it is shown that antisense mRNA of TGFbeta1 did block development of fibrosis. To state that the effect is on a TGFbeta-dependent activity, controlled 
TGFbeta-induced events have to be evaluated. 
4. Animal experiments were performed according to ¿strict governmental and international 
guidelines¿, which is a too general statement. Which guidelines were followed? 
5. Description of Figure 2, panels G and H, is very short. What do you mean as an obvious liver 
appearance? This is in contrast with the detailed description and presentation of a surgery procedure. 
6. Magnification bars in Figures 4 and 5 would be useful. 
Minor Compulsory Revisions (such as missing labels on figures, or the wrong use of a term, which 
the author can be trusted to correct) None 
Major Compulsory Revisions (that the author must respond to before a decision on publication can 
be reached) 
None 
 Accept after discretionary revisions Advice on publication: 
 A paper of considerable merit and interest in its field Level of interest: 
 Acceptable Quality of written English: 
Declaration of competing interests: 
None 
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