Reviewer ’s report 
 
Title: CTRC gene polymorphism (p.G60=; c.180 C>T) in acute pancreatitis  
 
Version: 1 Date: 05 Jul 2016  
 
Reviewer: Ulrich Weiss  
 
Reviewer's report:  
 
The manuscript by Koziel et al. is a genetic association study, investigating  the presence of the 
CTRC c.180C>T allele in Polish patients suffering from  acute pancreatitis and its potential 
influence on disease severity. The authors confirm and extend previous findings of higher allele 
frequencies in CP and recurrent AP also in their investigated cohort of acute pancreatitis. 
Surprisingly only heterozygous  carriers were more frequent in the AP cohort,  but not 
homozygous carrier. Also the authors claim that they found no significant correlation between 
having the CT or TT genotype and the severity of the clinical course of AP.  
 
 
Apparently this is a resubmission, answering several points of critique from a fist reviewing 
process. The study has interesting new findings and the topic is of clinical importance. Most of 
the reviewers questions seem to have been answered but there remain additional points that need 
to be addressed:  
 
 
 
1) To me the data support a r ole of the c.180C>T allele as a risk factor for developing acute 
pancreatitis. The findings also indicate, that the presence of the  c.180C>T allele is not 
associated with moderate or a severe course of AP (30.8% CT carrier vs. 31% in 
controls).  
 
However, I  would not support the notion that there is a difference between CT and 
homozygous carriers of the mutant allele. 31% of five homozygous patients means that 
only one or of them would have been expected to experience severe disease. The 
statement therefore depends on the presence or absence of a single patient, does not 
reflect a reliable finding and should be removed from the abstract.  
 
 
 
2) Abstract: "In the AP group in 1.7% of the cases, the gene CTRC mutation was revealed 
vs. 0.7% in the group of healthy pe ople".  
 
What CTRC mutation are you referring to ?  If you refer to additionally found CTRC 
mutations you should state that here otherwise the reader gets confused  ! In addition, no 
sequence analysis of the whole CTRC coding region has been performed with y our 
manuscript, at least it has not been described in your methods. It is very confusing to state 
results in the abstract which apparently are not part of the reported work.  Also, the role 
of other CTRC mutations is not the main topic of the present manus cript, so I would 
suggest to remove these statements from the paper. Otherwise the source of these data 
has to be clearly revealed and the analysis carefully explained  
 
 
 
3) Page 3 line 34: "cause of the disease is the activation of digestive enzymes in the 
follicular cells".  The pancreas has "acinar cells", but no follicular cells.  
 
 
 
4) Page 5 line 24: "Acute pancreatitis was diagnosed if a patient gained ≥10 points on the 
SADD Questionnaire, or if a period of alcohol abuse lasted ≥1 year, and the daily dose 
was 40 g of pure ethanol".  "A diagnosis of chronic pancreatitis was an exclusion 
criterion".  
 
This apparently is the diagnosis of acute "alcoholic pancreatitis not of AP in general ! 
What were the diagnostic criteria for chronic pancreatitis and the exclus ion of those 
patients ?  
 
 
 
5) Table 5 is very  confusing and should be reconsidered. Which specific SPINK1, CFTR, 
CTRC mutations  do the individuals carry. There are two different p -values in the table, 
but it is unclear how they have been calculated or what  they refer to. The description of 
these results (page 10)  is also rather vague and should be rewritten and more specific.  
Which analysis supports your conclusion that SPINK1 mutations in combination with the 
c.180C>T allele are responsible for a more se vere course of AP ?  
 
 
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
No 
 
Does the work include the neces sary controls?  
If not, please specify which controls are required in your comments to the authors.  
 
Yes 
 
Are the conclusions drawn adequately supported by the data shown?  
If not, please explain in your comments to the authors.  
 
No 
Are you able to asse ss any statistics in the manuscript or would you recommend an  
additional statistical review?  
If an additional statistical review is recommended, please specify what aspects require further  
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Acceptable  
 
 
 
Declaration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
 
1. Have you in the pa st five years received reimbursements, fees, funding, or salary from an  
organisation that may in any way gain or lose financially from the publication of this  
manuscript, either now or in the future?  
 
2. Do you hold any stocks or shares in an organisation t hat may in any way gain or lose  
financially from the publication of this manuscript, either now or in the future?  
 
3. Do you hold or are you currently applying for any patents relating to the content of the  
manuscript?  
 
4. Have you received reimbursements, f ees, funding, or salary from an organization that 
holds  or has applied for patents relating to the content of the manuscript?  
 
5. Do you have any other financial competing interests?  
 
6. Do you have any non -financial competing interests in relation to this p aper?  
 
 
If you can answer no to all of the above, write 'I declare that I have no competing interests' 
below.  If your reply is yes to any, please give details below.  
 
I declare that I have no competeing interests  
 
I agree to the open peer review policy of t he journal. I understand that my name will be included  
on my report to the authors and, if the manuscript is accepted for publication, my named report  
including any attachments I upload will be posted on the website along with the authors'  
responses. I agree for my report to be made available under a n Open Access Creative Commons  
CC-BY license ( http://creativecommons.org/licenses/by/4.0/ ). I understand that any comments  
which I do not wish to be included in my named report can be included as  confidential comments  
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
Reviewer ’s report 
 
Title: CTRC gene polymorphism (p.G60=; c.180 C>T) in acute pancreatitis  
 
Version: 1 Date: 05 Jul 2016  
 
Reviewer: Ulrich Weiss  
 
Reviewer's report:  
 
The manuscript by Koziel et al. is a genetic association study, investigating  the presence of the 
CTRC c.180C>T allele in Polish patients suffering from  acute pancreatitis and its potential 
influence on disease severity. The authors confirm and extend previous findings of higher allele 
frequencies in CP and recurrent AP also in their investigated cohort of acute pancreatitis. 
Surprisingly only heterozygous  carriers were more frequent in the AP cohort,  but not 
homozygous carrier. Also the authors claim that they found no significant correlation between 
having the CT or TT genotype and the severity of the clinical course of AP.  
 
 
Apparently this is a resubmission, answering several points of critique from a fist reviewing 
process. The study has interesting new findings and the topic is of clinical importance. Most of 
the reviewers questions seem to have been answered but there remain additional points that need 
to be addressed:  
 
 
 
1) To me the data support a r ole of the c.180C>T allele as a risk factor for developing acute 
pancreatitis. The findings also indicate, that the presence of the  c.180C>T allele is not 
associated with moderate or a severe course of AP (30.8% CT carrier vs. 31% in 
controls).  
 
However, I  would not support the notion that there is a difference between CT and 
homozygous carriers of the mutant allele. 31% of five homozygous patients means that 
only one or of them would have been expected to experience severe disease. The 
statement therefore depends on the presence or absence of a single patient, does not 
reflect a reliable finding and should be removed from the abstract.  
 
 
 
2) Abstract: "In the AP group in 1.7% of the cases, the gene CTRC mutation was revealed 
vs. 0.7% in the group of healthy pe ople".  
 
What CTRC mutation are you referring to ?  If you refer to additionally found CTRC 
mutations you should state that here otherwise the reader gets confused  ! In addition, no 
sequence analysis of the whole CTRC coding region has been performed with y our 
manuscript, at least it has not been described in your methods. It is very confusing to state 
results in the abstract which apparently are not part of the reported work.  Also, the role 
of other CTRC mutations is not the main topic of the present manus cript, so I would 
suggest to remove these statements from the paper. Otherwise the source of these data 
has to be clearly revealed and the analysis carefully explained  
 
 
 
3) Page 3 line 34: "cause of the disease is the activation of digestive enzymes in the 
follicular cells".  The pancreas has "acinar cells", but no follicular cells.  
 
 
 
4) Page 5 line 24: "Acute pancreatitis was diagnosed if a patient gained ≥10 points on the 
SADD Questionnaire, or if a period of alcohol abuse lasted ≥1 year, and the daily dose 
was 40 g of pure ethanol".  "A diagnosis of chronic pancreatitis was an exclusion 
criterion".  
 
This apparently is the diagnosis of acute "alcoholic pancreatitis not of AP in general ! 
What were the diagnostic criteria for chronic pancreatitis and the exclus ion of those 
patients ?  
 
 
 
5) Table 5 is very  confusing and should be reconsidered. Which specific SPINK1, CFTR, 
CTRC mutations  do the individuals carry. There are two different p -values in the table, 
but it is unclear how they have been calculated or what  they refer to. The description of 
these results (page 10)  is also rather vague and should be rewritten and more specific.  
Which analysis supports your conclusion that SPINK1 mutations in combination with the 
c.180C>T allele are responsible for a more se vere course of AP ?  
 
 
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
No 
 
Does the work include the neces sary controls?  
If not, please specify which controls are required in your comments to the authors.  
 
Yes 
 
Are the conclusions drawn adequately supported by the data shown?  
If not, please explain in your comments to the authors.  
 
No 
Are you able to asse ss any statistics in the manuscript or would you recommend an  
additional statistical review?  
If an additional statistical review is recommended, please specify what aspects require further  
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Acceptable  
 
 
 
Declaration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
 
1. Have you in the pa st five years received reimbursements, fees, funding, or salary from an  
organisation that may in any way gain or lose financially from the publication of this  
manuscript, either now or in the future?  
 
2. Do you hold any stocks or shares in an organisation t hat may in any way gain or lose  
financially from the publication of this manuscript, either now or in the future?  
 
3. Do you hold or are you currently applying for any patents relating to the content of the  
manuscript?  
 
4. Have you received reimbursements, f ees, funding, or salary from an organization that 
holds  or has applied for patents relating to the content of the manuscript?  
 
5. Do you have any other financial competing interests?  
 
6. Do you have any non -financial competing interests in relation to this p aper?  
 
 
If you can answer no to all of the above, write 'I declare that I have no competing interests' 
below.  If your reply is yes to any, please give details below.  
 
I declare that I have no competeing interests  
 
I agree to the open peer review policy of t he journal. I understand that my name will be included  
on my report to the authors and, if the manuscript is accepted for publication, my named report  
including any attachments I upload will be posted on the website along with the authors'  
responses. I agree for my report to be made available under a n Open Access Creative Commons  
CC-BY license ( http://creativecommons.org/licenses/by/4.0/ ). I understand that any comments  
which I do not wish to be included in my named report can be included as  confidential comments  
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
Please review this paper.
user
Reviewer ’s report 
 
Title: CTRC gene polymorphism (p.G60=; c.180 C>T) in acute pancreatitis  
 
Version: 1 Date: 05 Jul 2016  
 
Reviewer: Ulrich Weiss  
 
Reviewer's report:  
 
The manuscript by Koziel et al. is a genetic association study, investigating  the presence of the 
CTRC c.180C>T allele in Polish patients suffering from  acute pancreatitis and its potential 
influence on disease severity. The authors confirm and extend previous findings of higher allele 
frequencies in CP and recurrent AP also in their investigated cohort of acute pancreatitis. 
Surprisingly only heterozygous  carriers were more frequent in the AP cohort,  but not 
homozygous carrier. Also the authors claim that they found no significant correlation between 
having the CT or TT genotype and the severity of the clinical course of AP.  
 
 
Apparently this is a resubmission, answering several points of critique from a fist reviewing 
process. The study has interesting new findings and the topic is of clinical importance. Most of 
the reviewers questions seem to have been answered but there remain additional points that need 
to be addressed:  
 
 
 
1) To me the data support a r ole of the c.180C>T allele as a risk factor for developing acute 
pancreatitis. The findings also indicate, that the presence of the  c.180C>T allele is not 
associated with moderate or a severe course of AP (30.8% CT carrier vs. 31% in 
controls).  
 
However, I  would not support the notion that there is a difference between CT and 
homozygous carriers of the mutant allele. 31% of five homozygous patients means that 
only one or of them would have been expected to experience severe disease. The 
statement therefore depends on the presence or absence of a single patient, does not 
reflect a reliable finding and should be removed from the abstract.  
 
 
 
2) Abstract: "In the AP group in 1.7% of the cases, the gene CTRC mutation was revealed 
vs. 0.7% in the group of healthy pe ople".  
 
What CTRC mutation are you referring to ?  If you refer to additionally found CTRC 
mutations you should state that here otherwise the reader gets confused  ! In addition, no 
sequence analysis of the whole CTRC coding region has been performed with y our 
manuscript, at least it has not been described in your methods. It is very confusing to state 
results in the abstract which apparently are not part of the reported work.  Also, the role 
of other CTRC mutations is not the main topic of the present manus cript, so I would 
suggest to remove these statements from the paper. Otherwise the source of these data 
has to be clearly revealed and the analysis carefully explained  
 
 
 
3) Page 3 line 34: "cause of the disease is the activation of digestive enzymes in the 
follicular cells".  The pancreas has "acinar cells", but no follicular cells.  
 
 
 
4) Page 5 line 24: "Acute pancreatitis was diagnosed if a patient gained ≥10 points on the 
SADD Questionnaire, or if a period of alcohol abuse lasted ≥1 year, and the daily dose 
was 40 g of pure ethanol".  "A diagnosis of chronic pancreatitis was an exclusion 
criterion".  
 
This apparently is the diagnosis of acute "alcoholic pancreatitis not of AP in general ! 
What were the diagnostic criteria for chronic pancreatitis and the exclus ion of those 
patients ?  
 
 
 
5) Table 5 is very  confusing and should be reconsidered. Which specific SPINK1, CFTR, 
CTRC mutations  do the individuals carry. There are two different p -values in the table, 
but it is unclear how they have been calculated or what  they refer to. The description of 
these results (page 10)  is also rather vague and should be rewritten and more specific.  
Which analysis supports your conclusion that SPINK1 mutations in combination with the 
c.180C>T allele are responsible for a more se vere course of AP ?  
 
 
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
No 
 
Does the work include the neces sary controls?  
If not, please specify which controls are required in your comments to the authors.  
 
Yes 
 
Are the conclusions drawn adequately supported by the data shown?  
If not, please explain in your comments to the authors.  
 
No 
Are you able to asse ss any statistics in the manuscript or would you recommend an  
additional statistical review?  
If an additional statistical review is recommended, please specify what aspects require further  
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Acceptable  
 
 
 
Declaration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
 
1. Have you in the pa st five years received reimbursements, fees, funding, or salary from an  
organisation that may in any way gain or lose financially from the publication of this  
manuscript, either now or in the future?  
 
2. Do you hold any stocks or shares in an organisation t hat may in any way gain or lose  
financially from the publication of this manuscript, either now or in the future?  
 
3. Do you hold or are you currently applying for any patents relating to the content of the  
manuscript?  
 
4. Have you received reimbursements, f ees, funding, or salary from an organization that 
holds  or has applied for patents relating to the content of the manuscript?  
 
5. Do you have any other financial competing interests?  
 
6. Do you have any non -financial competing interests in relation to this p aper?  
 
 
If you can answer no to all of the above, write 'I declare that I have no competing interests' 
below.  If your reply is yes to any, please give details below.  
 
I declare that I have no competeing interests  
 
I agree to the open peer review policy of t he journal. I understand that my name will be included  
on my report to the authors and, if the manuscript is accepted for publication, my named report  
including any attachments I upload will be posted on the website along with the authors'  
responses. I agree for my report to be made available under a n Open Access Creative Commons  
CC-BY license ( http://creativecommons.org/licenses/by/4.0/ ). I understand that any comments  
which I do not wish to be included in my named report can be included as  confidential comments  
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
assistant