Reviewer ’s report  
 
Title: Intestinal Candida parapsilosis isolates from Rett syndrome subjects bear potential virulent 
traits  and capacity to persist within the host  
 
Version: 0 Date: 19 Dec 2017  
 
Reviewer: Roberto PIVA  
 
Reviewer's report:  
 
Strati et al. report the presence of putative virulent, pro -inflammatory intestinal C. parapsilosis 
strains as the most abundant yeast species in RTT subjects. The authors show that C. parapsilosis 
isolates from RTT subjects display distinct genotypic profiles and are significantly more resistant 
to azole antifungals as compared to healthy controls' isolates. The capacity of these intestinal 
fungi to persist within host is supported by the high levels of IL -10 produce d by PMBCs and 
mixed Th1/Th17 cells population induced by RTT C. parapsilosis.  
Altogether, the manuscript contains important aspects that should be considered in RTT -
assiociated gastrointestinal pathophysiology. The methods and techniques employed are 
appropriated.  
However, several criticisms should be addressed to improve the quality of the manuscript.  
 
Major comments  
1. Line 234 -244: the description of resistance to antifungals in HC and RTT subjects is clear.  
However, MICs reported in Figure 1 and Tabl e 1 are not concordant. The authors should 
provide correct data and specify in Figure 1 legend whether MIC90 values were used.  
 
2. Line 266 -269: the authors indicate that RTT isolates induced higher levels of IL -1b, IL -6, 
TNF -a, and IL -10. However, statisti cal analysis to support IL -6 and TNF -a increase is 
missing.  
 
3. Line 272: expression of IDO1 in PBMCs stimulated by C. parapsilosis isolates is not shown.  
 
Minor Comments  
1. The title should be adjusted to emphasize the link between RTT syndrome and C. 
parapsilosis.  
 
2. The abstract does not report the purpose of the paper; the background should synthesize and 
stress the link between RTT syndrome and fungal infection; the c onclusions are not very 
clear.  
 
3. Molecular mechanisms underlying pro -inflammatory status should be discussed more 
extensively.  
 
4. The paper is well structured and written; however it requires accurate grammar proof -
readings.  
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 contr ols? 
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.  
 
Yes 
 
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  
 
Quali ty of written English  
Please indicate 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 quest ions:  
 
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 o f 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 i nterests  
 
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 i n 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: Intestinal Candida parapsilosis isolates from Rett syndrome subjects bear potential virulent 
traits  and capacity to persist within the host  
 
Version: 0 Date: 19 Dec 2017  
 
Reviewer: Roberto PIVA  
 
Reviewer's report:  
 
Strati et al. report the presence of putative virulent, pro -inflammatory intestinal C. parapsilosis 
strains as the most abundant yeast species in RTT subjects. The authors show that C. parapsilosis 
isolates from RTT subjects display distinct genotypic profiles and are significantly more resistant 
to azole antifungals as compared to healthy controls' isolates. The capacity of these intestinal 
fungi to persist within host is supported by the high levels of IL -10 produce d by PMBCs and 
mixed Th1/Th17 cells population induced by RTT C. parapsilosis.  
Altogether, the manuscript contains important aspects that should be considered in RTT -
assiociated gastrointestinal pathophysiology. The methods and techniques employed are 
appropriated.  
However, several criticisms should be addressed to improve the quality of the manuscript.  
 
Major comments  
1. Line 234 -244: the description of resistance to antifungals in HC and RTT subjects is clear.  
However, MICs reported in Figure 1 and Tabl e 1 are not concordant. The authors should 
provide correct data and specify in Figure 1 legend whether MIC90 values were used.  
 
2. Line 266 -269: the authors indicate that RTT isolates induced higher levels of IL -1b, IL -6, 
TNF -a, and IL -10. However, statisti cal analysis to support IL -6 and TNF -a increase is 
missing.  
 
3. Line 272: expression of IDO1 in PBMCs stimulated by C. parapsilosis isolates is not shown.  
 
Minor Comments  
1. The title should be adjusted to emphasize the link between RTT syndrome and C. 
parapsilosis.  
 
2. The abstract does not report the purpose of the paper; the background should synthesize and 
stress the link between RTT syndrome and fungal infection; the c onclusions are not very 
clear.  
 
3. Molecular mechanisms underlying pro -inflammatory status should be discussed more 
extensively.  
 
4. The paper is well structured and written; however it requires accurate grammar proof -
readings.  
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 contr ols? 
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.  
 
Yes 
 
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  
 
Quali ty of written English  
Please indicate 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 quest ions:  
 
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 o f 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 i nterests  
 
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 i n 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: Intestinal Candida parapsilosis isolates from Rett syndrome subjects bear potential virulent 
traits  and capacity to persist within the host  
 
Version: 0 Date: 19 Dec 2017  
 
Reviewer: Roberto PIVA  
 
Reviewer's report:  
 
Strati et al. report the presence of putative virulent, pro -inflammatory intestinal C. parapsilosis 
strains as the most abundant yeast species in RTT subjects. The authors show that C. parapsilosis 
isolates from RTT subjects display distinct genotypic profiles and are significantly more resistant 
to azole antifungals as compared to healthy controls' isolates. The capacity of these intestinal 
fungi to persist within host is supported by the high levels of IL -10 produce d by PMBCs and 
mixed Th1/Th17 cells population induced by RTT C. parapsilosis.  
Altogether, the manuscript contains important aspects that should be considered in RTT -
assiociated gastrointestinal pathophysiology. The methods and techniques employed are 
appropriated.  
However, several criticisms should be addressed to improve the quality of the manuscript.  
 
Major comments  
1. Line 234 -244: the description of resistance to antifungals in HC and RTT subjects is clear.  
However, MICs reported in Figure 1 and Tabl e 1 are not concordant. The authors should 
provide correct data and specify in Figure 1 legend whether MIC90 values were used.  
 
2. Line 266 -269: the authors indicate that RTT isolates induced higher levels of IL -1b, IL -6, 
TNF -a, and IL -10. However, statisti cal analysis to support IL -6 and TNF -a increase is 
missing.  
 
3. Line 272: expression of IDO1 in PBMCs stimulated by C. parapsilosis isolates is not shown.  
 
Minor Comments  
1. The title should be adjusted to emphasize the link between RTT syndrome and C. 
parapsilosis.  
 
2. The abstract does not report the purpose of the paper; the background should synthesize and 
stress the link between RTT syndrome and fungal infection; the c onclusions are not very 
clear.  
 
3. Molecular mechanisms underlying pro -inflammatory status should be discussed more 
extensively.  
 
4. The paper is well structured and written; however it requires accurate grammar proof -
readings.  
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 contr ols? 
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.  
 
Yes 
 
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  
 
Quali ty of written English  
Please indicate 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 quest ions:  
 
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 o f 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 i nterests  
 
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 i n 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