Reviewer ’s report  
 
Title:  The utility of apoptosis inhibitor of macrophages as a possible diagnostic marker in 
patients with Crohn's dis ease 
 
Version: 0 Date: 28 Dec 2015 
 
Reviewer:  Edward  Barnes  
 
Reviewer’s report:  
 
Ono et al. present their findings regarding the use of apoptosis inhibitor of macrophages (AIM) 
as a diagnostic marker in the evaluation of patients with Inflammatory Bowel Disease (IBD), 
particularly Crohn's Disease (CD).  Several of the findings prese nted are quite interesting, 
including the apparent ability for AIM to differentiate CD from UC as well as other 
inflammatory intestinal conditions such as Behcet's Disease as well as the documentation of AIM 
present in staining of active macrophages within  the intestinal mucosa.  I think that the authors' 
conclusion that AIM may be useful as an "auxiliary method of discriminating between CD and 
other chronic intestinal diseases" is important, and the use of AIM as a novel biomarker in the 
further evaluation s that have been proposed seems worthwhile.  
 
I do have a few questions/comments from the manuscript that I would pose to the authors:  
 
1. On Page 8, Materials and Methods, Patients section:  "This study enrolled 107 IBD patients, 
39 of whom were diagnosed with CD and 51 with UC; 17 patients with intestinal Behcet's 
disease (BD) were used as intestinal disease controls (Table 1)." Although CD and Behcet's 
Disease can demonstrate similar clinical features, I would argue that most providers would still 
identi fy IBD and Behcet's Disease as two separate disease entities and as such I would not 
include these 17 patients into the total population of IBD patients.  I think that the comparison to 
patients with Behcet's disease makes sense as a control group, but wou ld list these as a separate 
entity and thus list only 90 patients total with IBD (51 UC, 39 CD).  
 
2. On Page 8, Materials and Methods, Patients section: "The active phase was defined as a CDAI 
score greater than or equal to 6."  As the CDAI >150 has alread y been defined a few lines above, 
perhaps this sentence was meant to refer to a UC index such as the Simple Clinical Colitis 
Activity Index (SCCAI)?  
 
3. How was the cutoff value for ASCA decided (320 U/mL)?  If this value was pre -determined as 
a cutoff bas ed on another study I would consider citing this study. However if this value was 
specific to your ROC analysis, I would consider an additional discussion of or comparison to 
more standard reported ASCA cutoff values, such as those used in the Prometheus l ab assay 
(17.8 EU/mL) for clarification.  The conclusion that under ROC analysis, AIM had better 
sensitivity and specificity than ASCA for the screening of CD would likely require further 
clarification for full acceptance and use a diagnostic marker.  
 
4. The expression of AIM in active macrophages in inflamed intestinal mucosa is an interesting 
finding.  Although there is some discussion of the potential reasons that AIM was not correlated 
with disease activity, are there any other hypothesized reasons for this lack of association?  This 
appears to be an area of planned future study by the authors.  
 
5. The final conclusion paragraph seems overreaching, considering that the use of AIM as a drug 
target is not discussed in other areas.  A summary conclusion tha t identifies that recognizes the 
identification of CD14 and CD16 positive cells and potential future directions is potentially 
appropriate.  However, the major aim of the study as stated in the abstract and introduction was 
to evaluate AIM kinetics in the blood and tissues of patients with IBD, and the utility of AIM as 
a biomarker of IBD.  As such, I would think that the conclusion more similar to the conclusion 
stated in the abstract "AIM is a novel biomarker of CD that can distinguish CD from UC or BD. 
It is suggested that AIM may contribute to intestinal inflammation by inhibiting the apoptosis of 
macrophages" would be appropriate.  
 
Overall, I found the potential use for AIM as a novel diagnostic biomarker among patients with 
CD intriguing and appreciate  the authors' efforts in this area.  
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
Yes 
 
Does the work include the necessary 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 assess 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 i ndicate the quality of language in the manuscript:  
 
Needs some language corrections before being published  
 
Declaration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
 
1. Have you in the past 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 that 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, fees, 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 paper?  
 
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 competing interests  
 
I agree to the open peer review policy of the 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 an  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 con fidential comments 
to the editors, which will not be published . 
 
I agree to the open peer review policy of the journal  
Reviewer ’s report  
 
Title:  The utility of apoptosis inhibitor of macrophages as a possible diagnostic marker in 
patients with Crohn's dis ease 
 
Version: 0 Date: 28 Dec 2015 
 
Reviewer:  Edward  Barnes  
 
Reviewer’s report:  
 
Ono et al. present their findings regarding the use of apoptosis inhibitor of macrophages (AIM) 
as a diagnostic marker in the evaluation of patients with Inflammatory Bowel Disease (IBD), 
particularly Crohn's Disease (CD).  Several of the findings prese nted are quite interesting, 
including the apparent ability for AIM to differentiate CD from UC as well as other 
inflammatory intestinal conditions such as Behcet's Disease as well as the documentation of AIM 
present in staining of active macrophages within  the intestinal mucosa.  I think that the authors' 
conclusion that AIM may be useful as an "auxiliary method of discriminating between CD and 
other chronic intestinal diseases" is important, and the use of AIM as a novel biomarker in the 
further evaluation s that have been proposed seems worthwhile.  
 
I do have a few questions/comments from the manuscript that I would pose to the authors:  
 
1. On Page 8, Materials and Methods, Patients section:  "This study enrolled 107 IBD patients, 
39 of whom were diagnosed with CD and 51 with UC; 17 patients with intestinal Behcet's 
disease (BD) were used as intestinal disease controls (Table 1)." Although CD and Behcet's 
Disease can demonstrate similar clinical features, I would argue that most providers would still 
identi fy IBD and Behcet's Disease as two separate disease entities and as such I would not 
include these 17 patients into the total population of IBD patients.  I think that the comparison to 
patients with Behcet's disease makes sense as a control group, but wou ld list these as a separate 
entity and thus list only 90 patients total with IBD (51 UC, 39 CD).  
 
2. On Page 8, Materials and Methods, Patients section: "The active phase was defined as a CDAI 
score greater than or equal to 6."  As the CDAI >150 has alread y been defined a few lines above, 
perhaps this sentence was meant to refer to a UC index such as the Simple Clinical Colitis 
Activity Index (SCCAI)?  
 
3. How was the cutoff value for ASCA decided (320 U/mL)?  If this value was pre -determined as 
a cutoff bas ed on another study I would consider citing this study. However if this value was 
specific to your ROC analysis, I would consider an additional discussion of or comparison to 
more standard reported ASCA cutoff values, such as those used in the Prometheus l ab assay 
(17.8 EU/mL) for clarification.  The conclusion that under ROC analysis, AIM had better 
sensitivity and specificity than ASCA for the screening of CD would likely require further 
clarification for full acceptance and use a diagnostic marker.  
 
4. The expression of AIM in active macrophages in inflamed intestinal mucosa is an interesting 
finding.  Although there is some discussion of the potential reasons that AIM was not correlated 
with disease activity, are there any other hypothesized reasons for this lack of association?  This 
appears to be an area of planned future study by the authors.  
 
5. The final conclusion paragraph seems overreaching, considering that the use of AIM as a drug 
target is not discussed in other areas.  A summary conclusion tha t identifies that recognizes the 
identification of CD14 and CD16 positive cells and potential future directions is potentially 
appropriate.  However, the major aim of the study as stated in the abstract and introduction was 
to evaluate AIM kinetics in the blood and tissues of patients with IBD, and the utility of AIM as 
a biomarker of IBD.  As such, I would think that the conclusion more similar to the conclusion 
stated in the abstract "AIM is a novel biomarker of CD that can distinguish CD from UC or BD. 
It is suggested that AIM may contribute to intestinal inflammation by inhibiting the apoptosis of 
macrophages" would be appropriate.  
 
Overall, I found the potential use for AIM as a novel diagnostic biomarker among patients with 
CD intriguing and appreciate  the authors' efforts in this area.  
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
Yes 
 
Does the work include the necessary 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 assess 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 i ndicate the quality of language in the manuscript:  
 
Needs some language corrections before being published  
 
Declaration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
 
1. Have you in the past 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 that 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, fees, 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 paper?  
 
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 competing interests  
 
I agree to the open peer review policy of the 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 an  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 con fidential 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:  The utility of apoptosis inhibitor of macrophages as a possible diagnostic marker in 
patients with Crohn's dis ease 
 
Version: 0 Date: 28 Dec 2015 
 
Reviewer:  Edward  Barnes  
 
Reviewer’s report:  
 
Ono et al. present their findings regarding the use of apoptosis inhibitor of macrophages (AIM) 
as a diagnostic marker in the evaluation of patients with Inflammatory Bowel Disease (IBD), 
particularly Crohn's Disease (CD).  Several of the findings prese nted are quite interesting, 
including the apparent ability for AIM to differentiate CD from UC as well as other 
inflammatory intestinal conditions such as Behcet's Disease as well as the documentation of AIM 
present in staining of active macrophages within  the intestinal mucosa.  I think that the authors' 
conclusion that AIM may be useful as an "auxiliary method of discriminating between CD and 
other chronic intestinal diseases" is important, and the use of AIM as a novel biomarker in the 
further evaluation s that have been proposed seems worthwhile.  
 
I do have a few questions/comments from the manuscript that I would pose to the authors:  
 
1. On Page 8, Materials and Methods, Patients section:  "This study enrolled 107 IBD patients, 
39 of whom were diagnosed with CD and 51 with UC; 17 patients with intestinal Behcet's 
disease (BD) were used as intestinal disease controls (Table 1)." Although CD and Behcet's 
Disease can demonstrate similar clinical features, I would argue that most providers would still 
identi fy IBD and Behcet's Disease as two separate disease entities and as such I would not 
include these 17 patients into the total population of IBD patients.  I think that the comparison to 
patients with Behcet's disease makes sense as a control group, but wou ld list these as a separate 
entity and thus list only 90 patients total with IBD (51 UC, 39 CD).  
 
2. On Page 8, Materials and Methods, Patients section: "The active phase was defined as a CDAI 
score greater than or equal to 6."  As the CDAI >150 has alread y been defined a few lines above, 
perhaps this sentence was meant to refer to a UC index such as the Simple Clinical Colitis 
Activity Index (SCCAI)?  
 
3. How was the cutoff value for ASCA decided (320 U/mL)?  If this value was pre -determined as 
a cutoff bas ed on another study I would consider citing this study. However if this value was 
specific to your ROC analysis, I would consider an additional discussion of or comparison to 
more standard reported ASCA cutoff values, such as those used in the Prometheus l ab assay 
(17.8 EU/mL) for clarification.  The conclusion that under ROC analysis, AIM had better 
sensitivity and specificity than ASCA for the screening of CD would likely require further 
clarification for full acceptance and use a diagnostic marker.  
 
4. The expression of AIM in active macrophages in inflamed intestinal mucosa is an interesting 
finding.  Although there is some discussion of the potential reasons that AIM was not correlated 
with disease activity, are there any other hypothesized reasons for this lack of association?  This 
appears to be an area of planned future study by the authors.  
 
5. The final conclusion paragraph seems overreaching, considering that the use of AIM as a drug 
target is not discussed in other areas.  A summary conclusion tha t identifies that recognizes the 
identification of CD14 and CD16 positive cells and potential future directions is potentially 
appropriate.  However, the major aim of the study as stated in the abstract and introduction was 
to evaluate AIM kinetics in the blood and tissues of patients with IBD, and the utility of AIM as 
a biomarker of IBD.  As such, I would think that the conclusion more similar to the conclusion 
stated in the abstract "AIM is a novel biomarker of CD that can distinguish CD from UC or BD. 
It is suggested that AIM may contribute to intestinal inflammation by inhibiting the apoptosis of 
macrophages" would be appropriate.  
 
Overall, I found the potential use for AIM as a novel diagnostic biomarker among patients with 
CD intriguing and appreciate  the authors' efforts in this area.  
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
Yes 
 
Does the work include the necessary 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 assess 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 i ndicate the quality of language in the manuscript:  
 
Needs some language corrections before being published  
 
Declaration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
 
1. Have you in the past 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 that 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, fees, 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 paper?  
 
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 competing interests  
 
I agree to the open peer review policy of the 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 an  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 con fidential comments 
to the editors, which will not be published . 
 
I agree to the open peer review policy of the journal  
assistant