PONE-D-21-03985

Effect of Malaria and HIV/AIDS co-infection on Red Blood Cell Indices and Its relation with the CD4 level of Patients on HAART in Bench Sheko Zone, Southwest Ethiopia.

PLOS ONE

Dear Dr. Solomon Ejigu

Thank you for submitting your manuscript to PLOS ONE. After careful review with two experts in the fields, we feel that your manuscripts but does not meet PLOS ONE’s publication criteria as it currently stands. You are welcome to submit a revised version of the manuscript for our consideration if you could rigorously address the all points raised during the review process. However, please aware a revision does not always lead to a publication.

Please address point by point for all reviewers' comments including expert reviewer#1's detailed points as well as expert reviewer#2's general points, which I agreed. In addition, 1) please use less abbreviations or define it clearly at the beginning; 2) please spell out the gender specific Anemia criteria used;  3) please address whether there is a potential inclusion bias between the groups as we did see obviously the coinfected group seems sicker as reflected by their CD4; and 4) please acknowledge the small number of population studied.

Please ensure that your decision is justified on PLOS ONE’s publication criteria and not, for example, on novelty or perceived impact.

For Lab, Study and Registered Report Protocols: These article types are not expected to include results but may include pilot data. 

Please submit your revised manuscript by 6/4/21. 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

We look forward to receiving your revised manuscript.

Kind regards,

Weijing He, M.D.

Academic Editor

PLOS ONE

Journal Requirements:

When submitting your revision, we need you to address these additional requirements.

1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at

https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and

https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf

2. In the Methods, please clarify that participants provided oral consent. Please also state in the Methods:

- Why written consent could not be obtained

- Whether the Institutional Review Board (IRB) approved use of oral consent

- How oral consent was documented

For more information, please see our guidelines for human subjects research: https://journals.plos.org/plosone/s/submission-guidelines#loc-human-subjects-research

3. Please provide a sample size and power calculation in the Methods, or discuss the reasons for not performing one before study initiation.

4. Please ensure you have discussed any potential limitations of your study in the Discussion, including study design, sample size and/or potential confounders.

5. Please include additional information regarding the survey or questionnaire used in the study and ensure that you have provided sufficient details that others could replicate the analyses. For instance, if you developed a questionnaire as part of this study and it is not under a copyright more restrictive than CC-BY, please include a copy, in both the original language and English, as Supporting Information.

6. PLOS requires an ORCID iD for the corresponding author in Editorial Manager on papers submitted after December 6th, 2016. Please ensure that you have an ORCID iD and that it is validated in Editorial Manager. To do this, go to ‘Update my Information’ (in the upper left-hand corner of the main menu), and click on the Fetch/Validate link next to the ORCID field. This will take you to the ORCID site and allow you to create a new iD or authenticate a pre-existing iD in Editorial Manager. Please see the following video for instructions on linking an ORCID iD to your Editorial Manager account: https://www.youtube.com/watch?v=_xcclfuvtxQ

7. We note that you have indicated that data from this study are available upon request. PLOS only allows data to be available upon request if there are legal or ethical restrictions on sharing data publicly. For information on unacceptable data access restrictions, please see http://journals.plos.org/plosone/s/data-availability#loc-unacceptable-data-access-restrictions.

In your revised cover letter, please address the following prompts:

a) If there are ethical or legal restrictions on sharing a de-identified data set, please explain them in detail (e.g., data contain potentially identifying or sensitive patient information) and who has imposed them (e.g., an ethics committee). Please also provide contact information for a data access committee, ethics committee, or other institutional body to which data requests may be sent.

b) If there are no restrictions, please upload the minimal anonymized data set necessary to replicate your study findings as either Supporting Information files or to a stable, public repository and provide us with the relevant URLs, DOIs, or accession numbers. Please see http://www.bmj.com/content/340/bmj.c181.long for guidelines on how to de-identify and prepare clinical data for publication. For a list of acceptable repositories, please see http://journals.plos.org/plosone/s/data-availability#loc-recommended-repositories.

We will update your Data Availability statement on your behalf to reflect the information you provide.

8. Your ethics statement should only appear in the Methods section of your manuscript. If your ethics statement is written in any section besides the Methods, please move it to the Methods section and delete it from any other section. Please ensure that your ethics statement is included in your manuscript, as the ethics statement entered into the online submission form will not be published alongside your manuscript.

[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: Yes

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: The manuscript titled "Effect of Malaria and HIV/AIDS co-infection on Red Blood Cell Indices and Its relationwith the CD4 level of Patients on HAART in Bench Sheko Zone, Southwest Ethiopia" by Solomon Ejigu et al explored Malaria and HIV co-infection and the effects on RBC of infected patients. This is a valid research. However, there is need for major corrections before manuscript can be accepted for publication. The manuscript lacks line numbering, hence it is difficult to make reference to sections of the document while reviewing it. The references are poorly done. I suggest that authors rewrite the entire references following the Journal's guidelines for references. The methods and results were also poorly presented and the discussion was not sequentially done. Please see other specific comments below:

Please correct the spelling of "co infected" to "co-infected" all through the manuscript. Do the same for co-infection.

Abstract:

Pg 2, line 5: "... HIV co infected patients were inconsistent." is this an inference made from the present study or a general setting in the study location? Please rephrase statement.

Pg 2, line 8: "in HIV patients’ co infected" should be "in HIV patients co-infected"

line 9: "and correlate these"

line 15: P ≤ 0.001? Please check and correct this. Use > or < to represent the significant difference and = for exact p value all through the manuscript.

line 16: "between CD4 count with MCV," please use appropriate grammar.

lines 16-18: "While in  MHC,  CD4  count  with  RBC  and  Hgb  and  in  HIV  without  malaria  CD4  count  with  MCV, MCH  and  MCHC  were  positively  correlated." This statement is not clear. Please recast for better understanding.

line 21: what is OH? Please write in full.

The entire result were not properly presented by the authors in the abstract. I suggest that authors use short, simple and clear sentences to describe the results for easy readability. For example, lines 19-21, authors wrote "in MHC was 63.4%, 61.3%  in female sex and 55.9% in patients with  CD4 count of ≤500 cells/μl." and went on to write "Anemia in MHC patients was  higher in those with CD4 count of ≤500  cells/μl (59.3%)" why are there two different results for the same parameter for MHC? (55.9% and 59.3%). Please revisit the result section of the abstract and represent findings in clear words.

line 26:" some  RBC  indices  in  total" please replace "in total" with a better phrase.

Introduction

Pg 3, line 1: "SSA". This is the first time this is being mentioned in the main body, please write in full before introducing acronym in parentheses. Do same for all similar cases (e.g. MHC in line 29, CBC and SOP in Pg 4, line 26 etc).

line 2: "due  to  a  number" please replace due to with "because"

line 8: please write 3/4th in words. P.  falciparum. Please write generic name in full as this is the first time it is mentioned in the manuscript.

line 11: Anopheles, Plasmodium

Pg 4. Authors have a section titled materials and methods, and another section titled methods. I suggest the removal of "methods" and division of the materials and methods into subsections including study location and population, study design, inclusion and exclusion criteria, sample collection, data analysis etc.

line 20: "(Biftu  and ". Where does the parentheses end? ")"

line 22: remove MHC from parentheses.

Results

Pg 5, line 5: "A  total  of  206  study  participants  complete  the  study." please rephrase statement.

line 6: "Female  were  53.9  %  totally  and  higher  in  both  MHC  and  OH," please recast statement.

line 7: "were  37.8  %  with  majority  (26  %)" I suggest maintaining 1 decimal place all through manuscript. Please use 26.0% instead.

lines 5-16: This section needs to be revised by a native English speaker. The manner in which the data were presented is confusing. E.g. I think it would be better understood if authors wrote "A total of 37.8% of the study participants were rural dwellers while 62.2% were urban settlers. 26.0% of the rural dwellers had MHC, while 38.4% of the urban dwellers had OH". Please consider consulting a native English speaker for proof-reading and thorough revision of manuscript.

Also, I suggest presenting results for the entire study population first, then each of the subgroups (MHC and OH) to avoid confusion. Alternatively, results can be presented as done on lines 10-11 "Totally  54.8%  had  CD4  count  of  ≥500  cells/μl  (22.8%  MHC  vs.  32 % OH)"

line 10: "P≤0.02" please refer to my previous comment on this. Secondly, what confidence interval was used for this study? 95% or 99%? Statistical difference should be presented as P < 0.05, P > 0.05, P < 0.01 or P > 0.01. 0.01 is 99% confidence interval, not 0.001. If authors wish to present the P value, then it should be presented with an "=", e.g. p = 0.048 on line 27.

Authors wrote "Totally  54.8%  had  CD4  count  of  ≥500  cells/μl  (22.8%  MHC  vs.  32 % OH)" on lines 10-11, and then went ahead to write "Majority  of  OH  (64.07%)  had  CD4  count  of  ≥500  cells/μl  as  compared  with  MHC  (45.6%)" on line 13. This is confusing. Please revisit results.

lines 24-28: "There  was  no  significant  correlation..." where was this result presented? Please indicate table/figure.

Pg 6, lines 3-4: "Anemia  in  female  was  higher  than in  male  (61.3  %  vs.38.7  %)  among  anemic." please rephrase statement.

lines 8-28: please specify the tables/figures where these results were presented.

Pages 7-9: Authors did not sequentially discuss the results from the study. I suggest that authors discuss the results starting with socio-demographic and clinical characteristics, RBC indices in MHC and OH, correlation of CD4 count and RBC Indices in MHC and OH, etc.

There are also a lot of grammatical errors in this section. e.g."... finding of Nigeria" "Study of Ghana" (line 10) "Study  from ..." (lines 14 and 16) etc

Reviewer #2: The study was performed to find the correlation of RBC Indices and CD4 count in only HIV-infected (OH) and Malaria HIV co-infected (MHC) patients. RBC’s indices of MHC patients were significantly lower as compared to OH patients. On the other hand, a positive correlation was found between MHC and OH patients with respect to anemia and CD4 count. In MHC patients, CD4 count was positively correlated with RBC and Hgb. While in OH patients, CD4 count was positively correlated with MCV, MCH, and MCHC. While these observations in part confirm previous studies, the RBC indices in MHC and OH in Bench Maji Zone in SSA remain to be clarified. I request the authors to re-write the article as it is challenging to follow and incoherent. Please consider dividing the materials and methods with subheadings such as Study subjects, Determination CD4 counts, Statistics, etc. Consider stating a clear hypothesis in the introduction. Please consider proofreading for grammar as the document is full of mistakes.

**********

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: Yes: Nneoma Confidence JeanStephanie Anyanwu

Reviewer #2: Yes: Himanshu Batra

[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.

Effect of Malaria and HIV/AIDS co-infection on Red Blood Cell Indices and Its relation with the CD4 level of Patients on HAART in Bench Sheko Zone, Southwest Ethiopia.

PONE-D-21-03985R1

Dear Dr.Solomon Ejigu ,

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,

Weijing He, M.D.

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

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: All comments have been addressed

**********

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: Yes

Reviewer #2: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: (No Response)

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: (No Response)

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: (No Response)

Reviewer #2: Thanks for addressing my comments during the unrest in your area. I sincerely appreciate it.

**********

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: Yes: Nneoma Confidence J Anyanwu (Ph.D)

Reviewer #2: Yes: Himanshu Batra