Association of SNPs within TMPRSS6 and BMP2 genes with iron deficiency status in Saudi Arabia

Osama M. Al-Amer; Atif Abdulwahab A. Oyouni; Mohammed Ali Alshehri; Abdulrahman Alasmari; Othman R. Alzahrani; Saad Ali S. Aljohani; Noura Alasmael; Abdulrahman Theyab; Mohammad Algahtani; Hadeel Al Sadoun; Khalaf F. Alsharif; Abdullah Hamad; Wed A. Abdali; Yousef MohammedRabaa Hawasawi

doi:10.1371/journal.pone.0257895

Peer Review History

Original SubmissionMarch 10, 2021
10 Mar 2021 Author Response Attachments Attachment Submitted filename: English proofreading letter.docx https://doi.org/10.1371/journal.pone.0257895.r001
7 May 2021 Decision Letter - Cinzia Ciccacci, Editor PONE-D-21-07480 Association of polymorphisms of genes BMP2 and TMPRSS6 with iron deficiency status in Saudi Arabia PLOS ONE Dear Dr. Hawsawi, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. Please pay particular attention to the criticisms raised by Reviewer2, including the problem with the English and the issue of genotype frequency discrepancy. Please submit your revised manuscript by Jun 19 2021 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. Please include the following items when submitting your revised manuscript: A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter. 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Please revise the manuscript to rephrase the duplicated text, cite your sources, and provide details as to how the current manuscript advances on previous work. Please note that further consideration is dependent on the submission of a manuscript that addresses these concerns about the overlap in text with published work. We will carefully review your manuscript upon resubmission, so please ensure that your revision is thorough. [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: Partly Reviewer #2: Partly ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes ****** 3. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: No Reviewer #2: Yes ****** 4. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: No Reviewer #2: No ****** 5. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: Sample size is so small for PhD paper On which criteria you classified the students as healthy and iron deficient Have you made any questionnaire for participants? If yes provide it Provide consent form of participants for provision of blood samples Primers used for PCR are previously used or novelty designed What is m in the first line of discussion? Provide the form of ethical committee or institution review board of university Reviewer #2: The current study tried to assess the effects of TMPRSS6 and BMP2 SNP on the risk of iron deficiency anaemia in adult females from Saudi Arabia. The authors have done a great job in planning and executing this study. Both the study design and the results are is clear laid out. However, the article would require a complete review of the English grammar to make it suitable for publication. Here are my comments on the article: 1. The title should be reviewed for correctness. For example, “polymorphisms of genes TMPRSS6 and BMP2” may be written as SNPs within TMPRSS6 and BMP2 genes with iron deficiency…. 2. Introduction is unnecessarily long. Can be shortened to begin by talking about IDA and exclude general description of iron. 3. Why was TMPRSS6 rs4820268 not included in the analysis. This SNP has been widely associated with low iron status in different global populations. 4. Further English editing might help to improve the grammar. For example, the last paragraph of the introduction “….to investigate polymorphism rs1421312 of the TMPRSS6 gene…” may be written as TMPRSS6 rs1421312. 5. In section 3.2, it is not clear what this 3-5cc means. It will be helpful to write this use SI units (mL for the blood volume). 6. In the results section, line 1, TMPRSS6 rs1421312 is wrongly written rs141312. 7. Genotype frequencies of the two SNPs in the study population differed widely from the global and all regional populations including Asian populations in the 1000 Genomes project. I recommend that the authors review their genotyping method to ensure that there were no errors. 8. In the section “Genotype distribution of TMPRSS6 rs1421312 and BMP2 rs235756 according to clinical parameters”, the description of the haematology and biochemical parameters using “normal” and “abnormal” should be high and low levels respectively. 9. In page 9, the sentence “Although the exact mechanism is unclear, compelling evidence emphasizes the role of TMPRSS6 polymorphisms in causing IDA”, needs to be reviewed. TMPRSS6 SNPs have been associated with IDA but causality has is not established. Similarly, the grammar in the first sentence of the last paragraph in page 9 needs to be reviewed. 10. In the last sentence on page 9, “TMPRSS6 rs141312” the SNP ID is incorrect. 11. The information about the location of BMP2 mentioned in the second sentence of page 10 needs to be reviewed. BMP2 is located in chromosome 20, see Here, but not Chr 2 as mentioned by the authors. 12. The last paragraph in page 10 needs to be reviewed. The sentence “we observed no significant association between increased risk for IDA and SNP rs235756 on BMP2” may be written as ….risk of IDA and BMP2 rs236756…. 13. The conclusion that the results of this study “can be used as a predictive marker of IDA is Sauda Arabia” is premature. Further research on more genetic markers is needed to verify the clinical usability of this findings. These two SNPs are not sufficient to make this conclusion. ****** 6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: No Reviewer #2: Yes:** Dr Momodou W. Jallow [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0257895.r002
Revision 1
17 Jul 2021 Author Response Letter of Response to Reviewer’s Comments Dear chief editor for PLOS ONE Journal Many thanks for your peer review and comments. With great pleasure, we would like to inform you that, the manuscript has been revised comprehensively and we address all the reviewers' comments. We hope our manuscript satisfied the editor and reviewer and be considered for publication in your decent journal. Our updated response to the reviewers as following: 1- Response to Reviewer #1: Sample size is so small for PhD paper Many thanks for the important comments and recommendations. In Saudi Arabia and due to the Arabian culture where females are totally separated from males in the university, it was very difficult to collect samples from the female students. We did our best to collect as much as we can and we finally obtained a total of 108 blood samples from female students. Moreover, for the iron deficient students, we excluded all students diagnosed with eating disorders, pregnant, and those taking nutritional medications or supplements which reduced the number of participants. On which criteria you classified the students as healthy and iron deficient Many thanks for the important comments and recommendations. The criteria for the students as healthy and iron deficient were according to the WHO iron deficiency anemia assessment which was published in 2001. Levels of ferritin < 15 ng/ml and haemoglobin < 12 g/dl were defined as iron deficiency anaemia, whereas levels of ferritin < 15 ng/ml and haemoglobin > 12 g/dl were defined as iron deficiency without anaemia. Have you made any questionnaire for participants? If yes provide it Many thanks for this essential comment. However, because we measured the serum iron level there was no questionnaire for the participants. Provide consent form of participants for provision of blood samples Many thanks for this crucial comment. Consent form of participants for provision of blood samples is attached as requested. Primers used for PCR are previously used or novelty designed Many thanks for this valuable point. The primers were novelty designed. What is m in the first line of discussion? Many thanks for this precise notice. The m was a mistake from the proofreading and it has been removed. The sentence is " was thought to be due to dietary and environmental factors" and it has been highlighted in yellow. Provide the form of ethical committee or institution review board of university. Many thanks for this essential point. Ethical approval (IRB #2018-36) is attached as requested. 2- Response to Reviewer #2: The current study tried to assess the effects of TMPRSS6 and BMP2 SNP on the risk of iron deficiency anaemia in adult females from Saudi Arabia. The authors have done a great job in planning and executing this study. Both the study design and the results are is clear laid out. However, the article would require a complete review of the English grammar to make it suitable for publication. Many thanks for this and the manuscript has been sent for proofreading and English grammar was reviewed. 1. The title should be reviewed for correctness. For example, “polymorphisms of genes TMPRSS6 and BMP2” may be written as SNPs within TMPRSS6 and BMP2 genes with iron deficiency…. Many thanks for this essential comment. The title has been changed according to your valuable advice, and highlighted in yellow. 2. Introduction is unnecessarily long. Can be shortened to begin by talking about IDA and exclude general description of iron. This is a valid point and we agree with the reviewer. Therefore, the introduction has been shortened and focused on IDA. 3. Why was TMPRSS6 rs4820268 not included in the analysis. This SNP has been widely associated with low iron status in different global populations. Many thanks for this valid point. In our previous publication, we studied the TMPRSS6 rs4820268 SNPs in details. 4. Further English editing might help to improve the grammar. For example, the last paragraph of the introduction “….to investigate polymorphism rs1421312 of the TMPRSS6 gene…” may be written as TMPRSS6 rs1421312. Many thanks for this point and the manuscript has been sent for proofreading and English grammar were reviewed. The mentioned sentence has been corrected and highlighted in yellow. 5. In section 3.2, it is not clear what this 3-5cc means. It will be helpful to write this use SI units (mL for the blood volume). Many thanks for this point and the 3-5cc was corrected to mL in the manuscript and highlighted in yellow in page 4. 6. In the results section, line 1, TMPRSS6 rs1421312 is wrongly written rs141312. Many thanks for this point. The SNP is now corrected as rs1421312 and highlighted in yellow. 7. Genotype frequencies of the two SNPs in the study population differed widely from the global and all regional populations including Asian populations in the 1000 Genomes project. I recommend that the authors review their genotyping method to ensure that there were no errors. Many thanks and the genotype frequencies of the two SNPs in the study population was reviewed and there was no error. 8. In the section “Genotype distribution of TMPRSS6 rs1421312 and BMP2 rs235756 according to clinical parameters”, the description of the haematology and biochemical parameters using “normal” and “abnormal” should be high and low levels respectively. Many thanks and the description of the haematology and biochemical parameters were changed and highlighted in yellow. 9. In page 9, the sentence “Although the exact mechanism is unclear, compelling evidence emphasizes the role of TMPRSS6 polymorphisms in causing IDA”, needs to be reviewed. TMPRSS6 SNPs have been associated with IDA but causality has is not established. Similarly, the grammar in the first sentence of the last paragraph in page 9 needs to be reviewed. Many thanks for this important point. The sentence was reviewed and corrected according to the reviewer advise and highlighted in yellow. 10. In the last sentence on page 9, “TMPRSS6 rs141312” the SNP ID is incorrect. Many thanks and the SNP has been corrected as rs1421312 and highlighted in yellow. 11. The information about the location of BMP2 mentioned in the second sentence of page 10 needs to be reviewed. BMP2 is located in chromosome 20, see Here, but not Chr 2 as mentioned by the authors. Many thanks for this important point, the zero was removed by mistake during the proofreading. Currently, it has been corrected to chromosome 20 and highlighted in yellow. 12. The last paragraph in page 10 needs to be reviewed. The sentence “we observed no significant association between increased risk for IDA and SNP rs235756 on BMP2” may be written as ….risk of IDA and BMP2 rs236756…. Many thanks for this important point and the sentence has been corrected accordingly. 13. The conclusion that the results of this study “can be used as a predictive marker of IDA is Sauda Arabia” is premature. Further research on more genetic markers is needed to verify the clinical usability of this findings. These two SNPs are not sufficient to make this conclusion. This is a valid point and we totally agree with the reviewer and therefore the statement has been removed. Many thanks for your fairly and timely peer review along with valuable comments. We addressed all the highlighted reviewers' comments with a hope that, the revised manuscript will be considered for publication in your decent journal. Looking forward to hearing from you soon. Best regards Yousef Hawsawi Attachments Attachment Submitted filename: Response to reviewer 17-05-2021.docx https://doi.org/10.1371/journal.pone.0257895.r003
2 Sep 2021 Decision Letter - Cinzia Ciccacci, Editor PONE-D-21-07480R1 Association of SNPs within TMPRSS6 and BMP2 genes with iron deficiency status in Saudi Arabia PLOS ONE Dear Dr. Hawsawi, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. ============================== Please, revise the manuscript taking into account the Reviewer 2 comments. ============================== Please submit your revised manuscript by Oct 17 2021 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. Please include the following items when submitting your revised manuscript: A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter. If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols. We look forward to receiving your revised manuscript. Kind regards, Cinzia Ciccacci Academic Editor PLOS ONE Journal Requirements: Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice. Additional Editor Comments (if provided): [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation. Reviewer #1: All comments have been addressed Reviewer #2: (No Response) ******** 2. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: Partly Reviewer #2: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes ****** 4. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: Yes Reviewer #2: Yes ****** 5. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: Yes Reviewer #2: Yes ****** 6. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: Thanks for addressing all concerns. Refrain dual publication of article. try to meet the criteria of sample size Reviewer #2: The authors have made significant improvements on the manuscript, and they have responded to the comments raised during the review. However, there are minor issues that needs to be addressed to improve the quality of the article as follows: 1. Abstract a. 3 – 5 cc in the methods section could be changes to 3 – 5mL. 2. Introduction section: a. The sentence “….developing and low-income countries” could be change to “low- and middle-income countries”. b. In line 3 of the third paragraph, the T in TMPRSS6 is not italised. 3. Results a. In the last sentence of the last paragraph, it is not clear what the authors meant by … “associated with recessive genotypes”. The phrase may be removed. 4. Discussion section a. The beginning of paragraph 3: “McLaren and his colleagues studied SNP in gene TMPRSS6 in four different ethnic groups…” should be revised. Also, the authors may consider removing the pronoun “his” when referring to other authors. Such sentences may be written as e.g. McLaren and colleagues studies. b. In paragraph 4, line 4, the “H” in HFE should be italised. c. Millet and his colleagues, see above comments d. In line 6, it is not necessary to include the P value 0.002 in discussion here and in other places. e. In paragraph 5, lines 4 – 5, the sentence “… direct association between TMPRSS6 and increase risk of IDA” should be revised. The association reported by these authors is between specific TMPRSS6 SNPs and IDA but not TMPRSS6 only. Additional general comments 1. Table 5 need reformatting. In the Odd ration column, there should be a space between the OR and the confidence interval e.g 3.74(0.19-73.05) should be written as 3.74 (0.19-73.05). 2. There should be a footnote to explain what “ref.” means 3. Table 6 should be on one page. And see the above comments regarding footnotes and brakets. ****** 7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: No Reviewer #2: Yes:** Momodou W. Jallow, PhD. [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0257895.r004
Revision 2
5 Sep 2021 Author Response Letter of Response to Reviewer’s Comments Dear chief editor for PLOS ONE Journal Many thanks for your distinguish peer review. With great pleasure, we would like to inform you that, the manuscript has been revised for the second round comprehensively and we address all the reviewers' comments. We hope our manuscript satisfied the editor and reviewers and be considered for publication in your decent journal. Our updated response to the reviewers as following: 1- Response to Reviewer #1: Reviewer #1: Thanks for addressing all concerns. Refrain dual publication of article. try to meet the criteria of sample size Many thanks for the positive comments and recommendations. We will try to meet the criteria of samples size in our next publication. We highly appreciate your valuable comments. 2- Response to Reviewer #2: Reviewer #2: The authors have made significant improvements on the manuscript, and they have responded to the comments raised during the review. Many thanks for the important comments and recommendations. It is highly appreciated. However, there are minor issues that need to be addressed to improve the quality of the article as follows: 1. Abstract a. 3 – 5 cc in the methods section could be changes to 3 – 5mL. Many thanks for this important point. The “3-5 cc” in the methods section has been changed to 3-5 mL. 2. Introduction section: a. The sentence “….developing and low-income countries” could be change to “low- and middle-income countries”. Many thanks for this important comments, the sentence has been changed to low- and middle-income countries. b. In line 3 of the third paragraph, the T in TMPRSS6 is not italised.. Many thanks for this precise comment and the T has been changed into italised. 3. Results a. In the last sentence of the last paragraph, it is not clear what the authors meant by … “associated with recessive genotypes”. The phrase may be removed. Many thanks for this essential comment. The phrase has been removed and the sentence changed to “associated with increased risk for IDA in the female students in the study”. 4. Discussion section a. The beginning of paragraph 3: “McLaren and his colleagues studied SNP in gene TMPRSS6 in four different ethnic groups…” should be revised. Also, the authors may consider removing the pronoun “his” when referring to other authors. Such sentences may be written as e.g. McLaren and colleagues studies. Many thanks for this important comment and the “McLaren and his colleagues studied SNP in gene TMPRSS6 in four different ethnic groups…” has been be revised and changed. The sentence McLaren and his colleagues has been corrected to McLaren and colleagues studies. b. In paragraph 4, line 4, the “H” in HFE should be italised. Many thanks for this precise comment and the H has been changed into italised and highlighted in yellow c. Millet and his colleagues, see above comments Many thanks for this important comment. The Millet and his colleagues has been changed to Millet and colleagues and highlighted in yellow. d. In line 6, it is not necessary to include the P value 0.002 in discussion here and in other places. Many thanks for this important comment, the P values were removed. e. In paragraph 5, lines 4 – 5, the sentence “… direct association between TMPRSS6 and increase risk of IDA” should be revised. The association reported by these authors is between specific TMPRSS6 SNPs and IDA but not TMPRSS6 only.. Many thanks for this important comment, the sentence has been corrected and the SNPS were included. 5. Additional general comments 1. Table 5 need reformatting. In the Odd ration column, there should be a space between the OR and the confidence interval e.g 3.74(0.19-73.05) should be written as 3.74 (0.19-73.05). The table has been space was added and the column was updated. 2. There should be a footnote to explain what “ref.” means The footnote was added. 3. Table 6 should be on one page. And see the above comments regarding footnotes and brakets. The footnote was added and the space also was added. Many thanks for the fairly and timely peer review along with valuable comments. We addressed all the highlighted reviewers' comments with a hope that, the revised manuscript will be considered for publication in your decent journal. Looking forward to hearing from you soon. Best regards Yousef Hawsawi Attachments Attachment Submitted filename: Response second to reviewer 05-09-2021.docx https://doi.org/10.1371/journal.pone.0257895.r005
14 Sep 2021 Decision Letter - Cinzia Ciccacci, Editor Association of SNPs within TMPRSS6 and BMP2 genes with iron deficiency status in Saudi Arabia PONE-D-21-07480R2 Dear Dr. Hawsawi, We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements. Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication. An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org. If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. Kind regards, Cinzia Ciccacci Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0257895.r006
Formally Accepted
5 Nov 2021 Acceptance Letter - Cinzia Ciccacci, Editor PONE-D-21-07480R2 Association of SNPs within TMPRSS6 and BMP2 genes with iron deficiency status in Saudi Arabia Dear Dr. Hawasawi: I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org. If we can help with anything else, please email us at plosone@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Dr. Cinzia Ciccacci Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0257895.r007

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