PONE-D-24-47138Application of Transthoracic Echocardiography to Assess the Dynamic Evolution of Early Cardiac Damage Induced by Abdominal Aorta-Inferior Vena Cava Fistula in Rats with Chronic Renal FailurePLOS ONE

Dear Dr. Wang,

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Academic Editor

PLOS ONE

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3. Thank you for stating the following financial disclosure: [This work was supported by : Natural Science Foundation of Shandong Province (ZR2023MH041)]. 

Please state what role the funders took in the study.  If the funders had no role, please state: ""The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.""

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Additional Editor Comments:

Major issues should be carefully addressed by the authors:

1) Title: add "...in female rats..." and discuss in the main text the choice of female rats.

2) "All surgery was performed under isoflurane...": please add concentration of the dose

3) Add information on assay used to measure each parameter.

4) Add reference supporting the CRF model and add more methodological details on the model.

5) Is echocardiography performed in awaken or anesthetized animals?

6) TOMTEC image analysis software: add details on the firm.

7) Table 1: Is the CRF model without significant changes in BUN and serum creatinine a clinically relevant model? Please discuss on it mentioning proper reference

8) Table 3: add data on heart rate and cardiac mass.

9) Echocardiographic parameters should be normalized on body weight

10) Figure legends: improve the text by adding sample size.

11) Since CRF leads to hypertrophic cardiomyocytes, the authors should measure size of cardiomyocytes. Finally, data should be discussed in the light of previous unmentioned study(10.1126/sciadv.adj4846).

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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: Yes

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

Reviewer #1: Yes

Reviewer #2: Yes

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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: Yes

Reviewer #2: Yes

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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: Yes

Reviewer #2: Yes

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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: Peer Review Report for Manuscript PONE-D-24-47138

1. Is the manuscript technically sound, and do the data support the conclusions?

The study presents a well-structured investigation into the use of transthoracic echocardiography for early detection of left ventricular dysfunction in a rat model of chronic renal failure with an arteriovenous fistula. The methodology is clearly outlined, and the experiments appear to have been conducted rigorously with appropriate controls. The statistical analysis is generally sound, and the data seem to support the conclusions. However, additional clarification on sample size justification and power analysis would strengthen the reliability of the findings.

One area of concern is the limited discussion of potential confounding factors, such as sex-based differences and possible variability in cardiac function assessments. Additionally, the inclusion of a more detailed comparison with existing literature could provide stronger contextual support for the study’s claims.

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

The manuscript employs standard statistical methods, including ANOVA and t-tests, to compare groups. However, while p-values are provided, effect sizes and confidence intervals should be reported to offer a clearer interpretation of the data’s clinical relevance. A justification for the sample size should be explicitly stated to confirm that the study is sufficiently powered to detect meaningful differences.

3. Have the authors made all data underlying the findings in their manuscript fully available?

The manuscript states that all relevant data are available within the article. However, to comply with PLOS ONE’s data-sharing policies, the authors should consider depositing raw data in a public repository and providing a DOI or accession number for transparency. Additionally, figures and tables should include access to numerical datasets where applicable.

4. Is the manuscript presented in an intelligible fashion and written in standard English?

The manuscript is well-written and generally clear. However, there are minor grammatical errors and awkward phrasing that should be addressed in revision. Some sections, particularly the results and discussion, would benefit from improved flow and organization to enhance readability. A thorough language edit is recommended to ensure clarity and precision.

Additional Comments:

Ethical considerations: The study adheres to animal research ethics guidelines, but the methods section should explicitly state whether humane endpoints were used to minimize animal suffering.

Publication ethics: No concerns about dual publication or research misconduct were identified.

Figures and tables: Some figure legends require more detailed explanations to ensure the data are easily interpretable.

Recommendation:

The study presents valuable insights into early cardiac dysfunction detection using echocardiography in a rat model of chronic renal failure. With improvements in statistical reporting, language clarity, and data transparency, the manuscript would be suitable for publication. Minor revisions are recommended.

Additional comments:

The manuscript should strictly follow the PLOS ONE submission guidelines, including structure, formatting, and citation style.

Experimental Design and Clarity:

The rationale for selecting female rats exclusively should be explained or justified, given potential sex-specific differences in cardiac remodeling.

The choice of a 6-week endpoint for the study is understandable due to mortality concerns, but it limits the ability to assess long-term changes. This limitation should be clearly emphasized in the discussion.

Data Presentation:

Figures and tables are critical for supporting key findings. Ensure all figures (e.g., histopathological images, strain imaging graphs) are high quality and provide enough detail to validate the claims. Some legends are unclear and should be revised for better comprehension.

Reviewer #2: This study presents a novel approach to assess left ventricular dysfunction (LVD) in rats with chronic renal failure (CRI) and arteriovenous fistula (AVF) using advanced imaging techniques. The researchers identified early functional changes by means of global longitudinal strain (GLS), global circumferential strain (GCS) and E/DT ratio, parameters that proved to be more sensitive than conventional echocardiography parameters. The results show continuous myocardial damage over time, underlining the potential of these innovative methods in detecting early cardiac dysfunction in a clinically relevant condition.

The study is well written and structured with adequate statistical analysis. Despite some limitations, the methodology is sound, and the conclusions are well justified, making this research a valuable contribution to the field.

Major comments:

- In the results better explain the results of EF and Strain, because we have an increase in EF and a decrease in Strain between the groups.

Minor Comments:

- Lines 123-123: 4 chamber view or 5 chamber view to obtain the LAD measurements.

- Lines 130-131: How do you obtain the RVCO? You said you don’t evaluate the tight heart for the artifact.

- In the tables will be useful to put the number of animals in each group.

- I suggest to put the values of the Systolic variables in the tables (LVESD, LVESV), the EF ad FS depends from them.

- Figure 2-3: the legend does not specify which group the bullette is from.

- For histopathological images, you have to put the same area for comparison. You have a section with the fibre in longitudinal axis and the comparison in transverse axis.

I suggest you improve the quality of the images.

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Reviewer #1: No

Reviewer #2: No

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Application of Transthoracic Echocardiography to Assess the Dynamic Evolution of Early Cardiac Damage Induced by Abdominal Aorta-Inferior Vena Cava Fistula in Rats with Chronic Renal Failure

PONE-D-24-47138R1

Dear Dr. Wang,

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.

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Kind regards,

Vincenzo Lionetti, M.D., PhD

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 #2: (No Response)

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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 #2: Yes

**********

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

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 #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 #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 #2: (No Response)

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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 #2: No

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