Reviewer's report 
 Caerulein-Induced Acute Pancreatitis in Mice that Constitutively Overexpress Reg/PAP Genes Title: 
 1  14 January 2006 Version: Date: 
 Makoto Otsuki Reviewer: 
Reviewer's report: 
General 
The pancreas of the CFTR null mouse (CF mouse) has elevated expression of the Reg/PAP genes 
and these genes are suggested to be involved in protection or recovery from pancreatic injury. 
Authors postulated that the elevated expression of Reg/PAP genes in the CF mouse pancreas 
ameliorates caerulein-induced pancreatitis. However, caerulein-induced pancreatitis in the CF mice 
was similar to or slightly severer than that in wild mice. Authors have suggested that elevated 
expression of the Reg/PAP genes in the CF mouse reduces apoptosis and increases the severity of 
pancreatitis. 
------------------------------------------------------------------------------- 
Major Compulsory Revisions (that the author must respond to before a decision on publication can 
be reached) 1. The Discussion should be shortened by at least 30%. 
2. Although authors stated that the severity of pancreatitis was increased by an elevation of the PAP 
gene expression and a reduction in apoptosis, it is not convincing because of the following two 
points. 
1) Although Reg3a proteins in the CF mice were significantly increased at 1h and 7h after induction 
of pancreatitis compared with those in wild type mice, significant increases in pancreatic MPO 
activities were observed only at 1h. 
2) The pancreas of the CF mouse is only mildly affected by loss of CFTR function. Since, however, there is already a mild inflammation in the pancreas as authors mentioned in the discussion, it is 
quite conceivable that the severity of pancreatitis in the CF mouse increased after repeated 
caerulein injections. 
3. Real time PCR was normalized against total RNA. Although total RNA was intact by gel 
electrophoresis, there is a possibility that total RNA is altered during pancreatitis. 
4. Authors suggest circulating Reg3 proteins contribute to airway inflammation. Although pancreatic 
Reg3 protein levels at 7h was the maximum, lung MPO levels at 7h and 12h in the CF mice were 
similar to those at control. How do you explain these difference between Reg3 protein and MPO 
levels? 
5. Many parameters were normalized at 7d after induction of pancreatitis in this report. Authors have 
to explain the reason why % wet weights at 7d in both mice significantly decreased compared to 
those at respective normal levels. 
6. Authors have to explain the reason why Reg3a protein levels remained unchanged during 
pancreatitis in both mice despite Reg3a mRNA levels were highly expressed. 
------------------------------------------------------------------------------- Minor Essential Revisions (such as missing labels on figures, or the wrong use of a term, which the author can be trusted to correct) 
1. Citation of reference is incorrect. The report of reference (9) (p3 line 10) demonstrates that Itmap1 
deficiency induces apoptosis and worsens severity of pancreatitis. 
2. Page 13 line 11: Fig. 10K; Fig. 8K. 
3. Page 18 line 21: The definite disparity is not clear. 
------------------------------------------------------------------------------- 
Discretionary Revisions (which the author can choose to ignore) 
 Accept after minor essential revisions What next?: 
 An article whose findings are important to those with closely related research Level of interest: 
interests 
 Acceptable Quality of written English: 
 No Statistical review: 
Declaration of competing interests: 
I declare that I have no competing interests. 
Reviewer's report 
 Caerulein-Induced Acute Pancreatitis in Mice that Constitutively Overexpress Reg/PAP Genes Title: 
 1  14 January 2006 Version: Date: 
 Makoto Otsuki Reviewer: 
Reviewer's report: 
General 
The pancreas of the CFTR null mouse (CF mouse) has elevated expression of the Reg/PAP genes 
and these genes are suggested to be involved in protection or recovery from pancreatic injury. 
Authors postulated that the elevated expression of Reg/PAP genes in the CF mouse pancreas 
ameliorates caerulein-induced pancreatitis. However, caerulein-induced pancreatitis in the CF mice 
was similar to or slightly severer than that in wild mice. Authors have suggested that elevated 
expression of the Reg/PAP genes in the CF mouse reduces apoptosis and increases the severity of 
pancreatitis. 
------------------------------------------------------------------------------- 
Major Compulsory Revisions (that the author must respond to before a decision on publication can 
be reached) 1. The Discussion should be shortened by at least 30%. 
2. Although authors stated that the severity of pancreatitis was increased by an elevation of the PAP 
gene expression and a reduction in apoptosis, it is not convincing because of the following two 
points. 
1) Although Reg3a proteins in the CF mice were significantly increased at 1h and 7h after induction 
of pancreatitis compared with those in wild type mice, significant increases in pancreatic MPO 
activities were observed only at 1h. 
2) The pancreas of the CF mouse is only mildly affected by loss of CFTR function. Since, however, there is already a mild inflammation in the pancreas as authors mentioned in the discussion, it is 
quite conceivable that the severity of pancreatitis in the CF mouse increased after repeated 
caerulein injections. 
3. Real time PCR was normalized against total RNA. Although total RNA was intact by gel 
electrophoresis, there is a possibility that total RNA is altered during pancreatitis. 
4. Authors suggest circulating Reg3 proteins contribute to airway inflammation. Although pancreatic 
Reg3 protein levels at 7h was the maximum, lung MPO levels at 7h and 12h in the CF mice were 
similar to those at control. How do you explain these difference between Reg3 protein and MPO 
levels? 
5. Many parameters were normalized at 7d after induction of pancreatitis in this report. Authors have 
to explain the reason why % wet weights at 7d in both mice significantly decreased compared to 
those at respective normal levels. 
6. Authors have to explain the reason why Reg3a protein levels remained unchanged during 
pancreatitis in both mice despite Reg3a mRNA levels were highly expressed. 
------------------------------------------------------------------------------- Minor Essential Revisions (such as missing labels on figures, or the wrong use of a term, which the author can be trusted to correct) 
1. Citation of reference is incorrect. The report of reference (9) (p3 line 10) demonstrates that Itmap1 
deficiency induces apoptosis and worsens severity of pancreatitis. 
2. Page 13 line 11: Fig. 10K; Fig. 8K. 
3. Page 18 line 21: The definite disparity is not clear. 
------------------------------------------------------------------------------- 
Discretionary Revisions (which the author can choose to ignore) 
 Accept after minor essential revisions What next?: 
 An article whose findings are important to those with closely related research Level of interest: 
interests 
 Acceptable Quality of written English: 
 No Statistical review: 
Declaration of competing interests: 
I declare that I have no competing interests. 
Please review this paper.
user
Reviewer's report 
 Caerulein-Induced Acute Pancreatitis in Mice that Constitutively Overexpress Reg/PAP Genes Title: 
 1  14 January 2006 Version: Date: 
 Makoto Otsuki Reviewer: 
Reviewer's report: 
General 
The pancreas of the CFTR null mouse (CF mouse) has elevated expression of the Reg/PAP genes 
and these genes are suggested to be involved in protection or recovery from pancreatic injury. 
Authors postulated that the elevated expression of Reg/PAP genes in the CF mouse pancreas 
ameliorates caerulein-induced pancreatitis. However, caerulein-induced pancreatitis in the CF mice 
was similar to or slightly severer than that in wild mice. Authors have suggested that elevated 
expression of the Reg/PAP genes in the CF mouse reduces apoptosis and increases the severity of 
pancreatitis. 
------------------------------------------------------------------------------- 
Major Compulsory Revisions (that the author must respond to before a decision on publication can 
be reached) 1. The Discussion should be shortened by at least 30%. 
2. Although authors stated that the severity of pancreatitis was increased by an elevation of the PAP 
gene expression and a reduction in apoptosis, it is not convincing because of the following two 
points. 
1) Although Reg3a proteins in the CF mice were significantly increased at 1h and 7h after induction 
of pancreatitis compared with those in wild type mice, significant increases in pancreatic MPO 
activities were observed only at 1h. 
2) The pancreas of the CF mouse is only mildly affected by loss of CFTR function. Since, however, there is already a mild inflammation in the pancreas as authors mentioned in the discussion, it is 
quite conceivable that the severity of pancreatitis in the CF mouse increased after repeated 
caerulein injections. 
3. Real time PCR was normalized against total RNA. Although total RNA was intact by gel 
electrophoresis, there is a possibility that total RNA is altered during pancreatitis. 
4. Authors suggest circulating Reg3 proteins contribute to airway inflammation. Although pancreatic 
Reg3 protein levels at 7h was the maximum, lung MPO levels at 7h and 12h in the CF mice were 
similar to those at control. How do you explain these difference between Reg3 protein and MPO 
levels? 
5. Many parameters were normalized at 7d after induction of pancreatitis in this report. Authors have 
to explain the reason why % wet weights at 7d in both mice significantly decreased compared to 
those at respective normal levels. 
6. Authors have to explain the reason why Reg3a protein levels remained unchanged during 
pancreatitis in both mice despite Reg3a mRNA levels were highly expressed. 
------------------------------------------------------------------------------- Minor Essential Revisions (such as missing labels on figures, or the wrong use of a term, which the author can be trusted to correct) 
1. Citation of reference is incorrect. The report of reference (9) (p3 line 10) demonstrates that Itmap1 
deficiency induces apoptosis and worsens severity of pancreatitis. 
2. Page 13 line 11: Fig. 10K; Fig. 8K. 
3. Page 18 line 21: The definite disparity is not clear. 
------------------------------------------------------------------------------- 
Discretionary Revisions (which the author can choose to ignore) 
 Accept after minor essential revisions What next?: 
 An article whose findings are important to those with closely related research Level of interest: 
interests 
 Acceptable Quality of written English: 
 No Statistical review: 
Declaration of competing interests: 
I declare that I have no competing interests. 
assistant