Verified Rigor RecordRigor Reviewed

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Circ_0081054 facilitates melanoma development via sponging miR‐637 and regulating RAB9A · 10.1111/srt.13313

Name: Verified Rigor Record — Circ_0081054 facilitates melanoma development via sponging miR‐637 and regulating RAB9A · 10.1111/srt.13313
Item: Circ_0081054 facilitates melanoma development via sponging miR‐637 and regulating RAB9A · 10.1111/srt.13313
Rating: 71
Author: Αlpha¹

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Αlpha¹ Editorial

This rigor review was examined and confirmed by Αlpha¹ Editorial · July 6, 2026

Αlpha¹ Rigor Review

Rigor Reviewed

WARN

Warnings to address · Interventional Study

Eight dimensions — click a segment for details

Critical

High-priority

Model-reported confidence: highReviewer agreement: moderateEngine: 7.6.0Source: full text

Not verified in this report

No reported statistical tests were found to recompute.

This Αlpha¹ Rigor Review uses AI Rigor Reviewers trained on a curated corpus of high-fidelity and retracted papers, with expert supervision and curation. It can still make mistakes; verify each finding against the source before relying on it.

The paper has clear ethical approvals and a logical scientific premise, but suffers from critical deficiencies in statistical reporting (only threshold p-values, no effect sizes), a factually incorrect data availability statement, and incomplete rigor details (no blinding, no power analysis, no cell line authentication). The statistical analysis and data availability are the most consequential weaknesses.

Three independent reviewers evaluated the full-text manuscript and all flagged similar issues. The statistics component could not recompute any tests because only threshold p-values were reported (coverage note). The citation component found 12 references not found in Crossref/OpenAlex, raising fabrication concerns, though none were confirmed retracted.

12 major claims checked against the paper's own evidence: all adequately supported.

8 integrity concerns flagged (1 high).

Rigor

Eight Dimensions2 pass · 5 warn · 1 fail

12 major claims2 partial10 supported

The introduction cites multiple prior studies on circRNAs in melanoma and notes that the role of circ_0081054 is unclear, establishing a logical premise. Limitations of prior research are not explicitly addressed as a separate point, but the premise rational is clear.

Sub-criteria

prior work citedADEQUATE

premise rationaleADEQUATE

limitations addressedNOT REPORTED

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 90% conf

The paper adequately cites prior work on circRNA in melanoma and establishes a clear rationale for investigating circ_0081054.

Evidence

direct quote[Introduction, paragraphs 3-4]

“For example, circ_0062270 regulated miR‐331‐3p/EPHA2 axis thus promoted melanoma development.”

direct quote[Introduction, paragraph 4]

“Circ_0081054 was upregulated in melanoma tissues, but, the role and regulatory mechanism of circ_0081054 in melanoma is still unclear.”

absence[Introduction]

Limitations of prior work not explicitly discussed.

PASSReviewer 2· 90% conf

The paper establishes a clear scientific premise by citing prior work on circRNAs in melanoma, providing a rationale for the study, and acknowledging some gaps.

Evidence

paraphrase[Introduction, paragraph 2]

“Cites examples such as circ_0062270, circ_0084043, and circ_0016418 in melanoma.”

direct quote[Introduction, paragraph 2]

“Circ_0081054 was upregulated in melanoma tissues, but, the role and regulatory mechanism of circ_0081054 in melanoma is still unclear.”

absence[Introduction, paragraph 2]

Limitations of prior research are barely addressed.

PASSReviewer 3· 90% conf

The paper adequately cites prior work and establishes a logical rationale linking circ_0081054, miR-637, and RAB9A to melanoma, though limitations of prior research are addressed only indirectly.

Evidence

direct quote[Introduction, paragraph 5]

“For example, circ_0062270 regulated miR-331-3p/EPHA2 axis thus promoted melanoma development.”

paraphrase[Introduction, final paragraph]

“The study states that it aims to measure dysregulations and biological roles of circ_0081054, miR-637 and RAB9A, and to validate the ceRNA network.”

direct quote[Discussion, paragraph 2]

“However, the in vivo experiment only investigated the expression of miR-637 and RAB9A in the primary tumors from A375 cells.”

Randomization and blinding are not reported for cell culture or animal experiments. No a priori power analysis is provided. Replicate distinction is noted as 'three repeats of cell experiments', but it is unclear if these are biological or technical replicates. Inclusion/exclusion criteria and outlier handling are not described. Controls (si-con, miR-con, pcDNA) are used but not discussed as formal positive/negative controls. No independent replication is reported.

Sub-criteria

controlsINADEQUATE

power analysisNOT REPORTED

blinding levelsNOT REPORTED

outlier handlingNOT REPORTED

randomization unitNOT REPORTED

inclusion exclusionNOT REPORTED

randomization methodNOT REPORTED

replicate distinctionNOT REPORTED

independent replicationNOT REPORTED

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 80% conf

The design lacks several key rigor elements: no blinding, no power analysis, no distinction between biological and technical replicates, and no pre-specified inclusion/exclusion or outlier handling for in vitro experiments.

Evidence

absence[Methods]

No blinding is described.

absence[Methods]

Power analysis is absent.

direct quote[Statistical analysis, Methods]

“There were three repeats of cell experiments.”

WARNReviewer 2· 75% conf

The study design lacks several key rigor elements: no randomization or blinding, no power analysis, no distinction between biological and technical replicates, and no explicit outlier handling. Inclusion/exclusion criteria and controls are partially reported.

Evidence

direct quote[Methods, section 2.6]

“Two small interfering RNAs (siRNA) against circ_0081054 (si-circ_0081054 #1 and si-circ_0081054 #2 ) and matched control (si-con)...”

direct quote[Methods, section 2.18]

“There were three repeats of cell experiments.”

absence[Methods, section 2]

No randomization or blinding is reported.

WARNReviewer 3· 75% conf

The study design includes multiple experimental approaches, but lacks randomization, blinding, power analysis, and clear replicate distinction for the in vitro experiments, and the animal study has limited design details.

Evidence

direct quote[Methods, section 2.18]

“There were three repeats of cell experiments.”

absence[Methods, section 2.17]

No randomization method or blinding is mentioned for the in vivo study.

direct quote[Methods, section 2.6]

“Two small interfering RNAs (siRNA) against circ_0081054 (si-circ_0081054 #1 and si-circ_0081054 #2 ) and matched control (si-con)...”

Animal sex (male) and age (5-week-old) are reported, but weight is not. Species/strain is given as 'BALB/C mice' but the substrain (e.g., BALB/c nu/nu) is not specified; source (SLAC) is provided. Housing conditions are briefly described as 'pathogen-free environment' but no details on cycle/temperature/enrichment. For cell lines, source and species are given but no biological variables (e.g., sex of donor, passage number) are reported. Health status is not mentioned for cells.

Sub-criteria

demographicsN/A

sex reportedADEQUATE

sex justifiedN/A

age weight healthINADEQUATE

housing conditionsINADEQUATE

species strain sourceINADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 85% conf

Sex and age/weight/health are reported for animals, but species/strain/source are incomplete, and housing conditions are vaguely described. Cell lines are identified but no biological variables are reported for them.

Evidence

direct quote[Methods, Tumor xenograft assay]

“A total of 18 male BALB/C mice (male; 5‐week‐old)”

direct quote[Methods, Tumor xenograft assay]

“were kept in pathogen‐free environment.”

direct quote[Methods, Cell lines]

“Human epidermal melanocyte (HEM) was bought from ScienCell... Melanoma cell lines A375 and A2058 were bought from American Type Culture Collection”

PASSReviewer 2· 85% conf

Sex (male) and age (5-week-old) of animals are reported, and human cell lines are identified. However, housing conditions are only partially described and health status is not mentioned.

Evidence

direct quote[Methods, section 2.17]

“A total of 18 male BALB/C mice (male; 5-week-old) purchased from SLAC (Shanghai, China)”

direct quote[Methods, section 2.17]

“kept in pathogen-free environment”

direct quote[Methods, section 2.2]

“Human epidermal melanocyte (HEM) was bought from ScienCell...Melanoma cell lines A375 and A2058 were bought from American Type Culture Collection (ATCC, Manassas, VA, USA).”

WARNReviewer 3· 70% conf

Sex of mice is reported, but age/weight are only partially given; strain and source are adequate; housing conditions are vague; human demographics are not reported.

Evidence

direct quote[Methods, section 2.17]

“A total of 18 male BALB/C mice (male; 5-week-old) purchased from SLAC (Shanghai, China) were kept in pathogen-free environment.”

absence[Methods, section 2.1]

Human demographics (age, sex, comorbidities) are not reported for the 52 melanoma patients.

direct quote[Methods, section 2.17]

“were kept in pathogen-free environment.”

The human tissue study was approved by the 'Ethics Committee of the Affiliated Huaian NO. 1 People's Hospital of Nanjing Medical University' and written informed consent was obtained from all participants. The animal study was approved by the 'Animal Care and Use Committee of the Affiliated Huaian NO.1 People's Hospital of Nanjing Medical University'. Regulatory compliance is implicitly addressed through the approval statements.

Sub-criteria

iacuc statementADEQUATE

informed consentADEQUATE

irb ethics statementADEQUATE

regulatory complianceINADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 95% conf

Ethics approval is explicitly stated for both human sample collection and the animal study, with named committees and informed consent for human participants.

Evidence

direct quote[Methods, Sample collection]

“This research was conducted with the approval of the Ethics Committee of the Affiliated Huaian NO. 1 People's Hospital of Nanjing Medical University.”

direct quote[Methods, Sample collection]

“All patients provided informed consent before starting this study.”

direct quote[Methods, Tumor xenograft assay]

“Animal study was approved by the Animal Care and Use Committee of the Affiliated Huaian NO.1 People's Hospital of Nanjing Medical University.”

PASSReviewer 2· 90% conf

Ethics approval for human tissue collection and animal experiments is reported with named committees. Informed consent from patients is stated.

Evidence

direct quote[Methods, section 2.1]

“This research was conducted with the approval of the Ethics Committee of the Affiliated Huaian NO. 1 People's Hospital of Nanjing Medical University.”

direct quote[Methods, section 2.1]

“All patients provided informed consent before starting this study.”

direct quote[Methods, section 2.17]

“Animal study was approved by the Animal Care and Use Committee of the Affiliated Huaian NO.1 People's Hospital of Nanjing Medical University.”

PASSReviewer 3· 90% conf

Ethical approvals for both human tissue collection and the animal study are reported with named committees, and informed consent is stated.

Evidence

direct quote[Methods, section 2.1]

“This research was conducted with the approval of the Ethics Committee of the Affiliated Huaian NO. 1 People's Hospital of Nanjing Medical University.”

direct quote[Methods, section 2.17]

“Animal study was approved by the Animal Care and Use Committee of the Affiliated Huaian NO.1 People's Hospital of Nanjing Medical University.”

direct quote[Methods, section 2.1]

“All patients provided informed consent before starting this study.”

Antibodies from Abcam are listed with catalog numbers and dilutions. Cell lines are sourced from commercial vendors but no authentication (STR profiling) or mycoplasma testing is stated. The xenograft uses BALB/C mice from SLAC with no genotyping details. Key reagents (TRIzol, kits) are identified with vendors. Software like SPSS 22.0 is named with version; ImageJ is used but version is not stated.

Sub-criteria

mycoplasma testingNOT REPORTED

reagents identifiedADEQUATE

organisms identifiedINADEQUATE

antibodies identifiedADEQUATE

cell line authenticationNOT REPORTED

software tools identifiedINADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 80% conf

Antibodies are identified with catalog numbers and applications, but cell line authentication and mycoplasma testing are not reported. Reagents are adequately identified, and software is identified but versions are sometimes missing.

Evidence

direct quote[Methods, Western blot analysis]

“The antibodies (Abcam, Cambridge, UK) including: hexokinase II (HK2; ab227198, 1:1000), β‐actin (ab8227, 1:5,000)...”

absence[Methods]

Cell line authentication and mycoplasma testing not reported.

direct quote[Methods, Statistical analysis]

“SPSS 22.0 (SPSS Inc., Chicago, IL, USA)”

WARNReviewer 2· 80% conf

Antibodies are identified with catalog numbers, cell lines have sources, reagents are mostly identified, and software is named. However, cell line authentication and mycoplasma testing are not reported, and primer sequences are provided in a table but some details are lacking.

Evidence

direct quote[Methods, section 2.11]

“The antibodies (Abcam, Cambridge, UK) including: hexokinase II (HK2; ab227198, 1:1000), β-actin (ab8227, 1:5,000), Goat Anti-Rabbit IgG H&L secondary antibody (ab205718, 1:3000), RAB9A (ab235538, 1:2000).”

absence[Methods, section 2.2]

Cell line authentication is not mentioned.

absence[Methods, section 2.2]

Mycoplasma testing is not mentioned.

WARNReviewer 3· 80% conf

Cell lines, antibodies, and reagents are identified with vendors and catalog numbers, but organisms (mice) are not given an RRID, custom primers are listed but not as RRIDs, and software (SPSS, ImageJ) is named without versions for ImageJ.

Evidence

direct quote[Methods, section 2.11]

paraphrase[Methods, section 2.2]

“Cell lines were bought from ATCC and ScienCell, but no authentication (e.g., STR profiling) or mycoplasma testing is reported.”

direct quote[Methods, section 2.9]

“Image J (NIH, Bethesda, Maryland, USA) software was used for invasive cells counting.”

The paper states 'Analysis of variance or Student's t-test' and Pearson correlation. Assumptions are not verified. p-values are only given as '* p < 0.05' in figures. Effect sizes and confidence intervals are not reported. SPSS 22.0 is identified. Data presentation uses bar graphs (as shown in figures) without individual data points. No arithmetic checks are possible as exact values are not reported in the text for key outcomes (e.g., mean ± SD for each group). mathematical_plausibility is not_applicable given limited numerical reporting.

Sub-criteria

tests namedINADEQUATE

exact p valuesNOT REPORTED

effect sizes ciNOT REPORTED

data presentationINADEQUATE

software identifiedADEQUATE

assumptions verifiedNOT REPORTED

mathematical plausibilityN/A

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 90% conf

The statistical analysis has multiple critical failures: no exact p-values are reported (only 'p < 0.05'), no effect sizes or confidence intervals, test assumptions are not verified, and a strong potential mathematical inconsistency is present when checking the provided sample size of 52 tissues against the reported p-value from a correlation of a single variable.

Evidence

direct quote[Throughout Results]

“* p < 0.05.”

absence[Results]

No effect sizes or CIs.

direct quote[Statistical analysis, Methods]

“Analysis of variance or Student's t‐test was used to analyze relevant data”

WARNReviewer 2· 70% conf

Statistical tests are named (t-test, ANOVA), and software (SPSS 22.0) is identified. However, exact p-values are not reported (only p < 0.05), effect sizes and confidence intervals are absent, and assumptions are not verified. Data presentation mostly shows bar graphs without individual points.

Evidence

direct quote[Methods, section 2.18]

“Analysis of variance or Student's t-test was used to analyze relevant data through SPSS 22.0”

direct quote[Results, throughout]

“* p < 0.05.”

absence[Methods, section 2.18]

Effect sizes and confidence intervals are not reported.

WARNReviewer 3· 80% conf

Statistical tests are named (t-test, ANOVA, Pearson correlation), but exact p-values are not reported (only p<0.05), effect sizes and confidence intervals are absent, software is partially identified, and data presentation mostly uses bar charts without individual points.

Evidence

direct quote[Methods, section 2.18]

“Analysis of variance or Student's t-test was used to analyze relevant data through SPSS 22.0”

direct quote[All figure legends]

“* p < 0.05.”

absence[Results sections 3.1-3.8]

No effect sizes or confidence intervals are provided.

The data availability statement 'Data sharing is not applicable to this article as no new data were created or analyzed in this study' is demonstrably false given the extensive new data reported. No data repository deposit, accession numbers, or code sharing are described. The statement is 'reported_but_inadequate' because it exists, but it misrepresents the study and provides no access route to the underlying data.

Sub-criteria

code sharingNOT REPORTED

accession numbersNOT REPORTED

repository depositNOT REPORTED

data availability statementINADEQUATE

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 95% conf

The data availability statement is contradictory and ultimately inadequate: it first says data sharing is not applicable as no new data were created, yet the paper clearly reports new experimental data from 52 melanoma tissues, cell lines, and in vivo experiments.

Evidence

direct quote[Data Availability Statement]

“Data sharing is not applicable to this article as no new data were created or analyzed in this study.”

absence[Data Availability Statement]

No data repository mentioned.

WARNReviewer 2· 95% conf

A data availability statement is present but contradictory: it says 'no new data were created,' yet the study generated experimental data from cell lines and animals. No repository deposit or code sharing is provided.

Evidence

direct quote[Data Availability Statement]

“Data sharing is not applicable to this article as no new data were created or analyzed in this study.”

absence[Data Availability Statement]

No repository deposit for raw data.

absence[Methods]

No code sharing.

FAILReviewer 3· 95% conf

The data availability statement is contradictory and inadequate: it claims no new data were created, but the study generated human tissue and cell line data. No data or code is deposited.

Evidence

direct quote[Data Availability Statement]

“Data sharing is not applicable to this article as no new data were created or analyzed in this study.”

absence[Entire paper]

No data repository deposit is reported.

absence[Entire paper]

No code sharing is reported.

Methods are detailed enough for replication. The paper is not a clinical trial and no registration is expected. No reporting guideline (e.g., ARRIVE) is referenced. Limitations are briefly noted (e.g., 'only investigated the expression of miR‐637 and RAB9A in the primary tumors from A375 cells'). Conclusions are appropriately cautious. Funding and COI are stated ('The authors received no specific funding for this work') but no grant numbers are applicable.

Sub-criteria

funding coiADEQUATE

trial registrationN/A

reporting guidelineNOT REPORTED

methods completenessADEQUATE

all outcomes reportedADEQUATE

limitations discussedINADEQUATE

conclusions proportionalADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 85% conf

Methods are largely complete, limitations are briefly discussed, and conclusions are proportional, but the paper is not pre-registered, no reporting guideline is followed, and funding/COI are stated but funding is explicitly absent.

Evidence

direct quote[Methods]

“Quantitative real‐time polymerase chain reaction (qRT‐PCR) assay was employed to evaluate RNA expression. Trizol reagent (Invitrogen, Carlsbad, CA, USA) was used isolate RNA...”

direct quote[Discussion]

“However, the in vivo experiment only investigated the expression of miR‐637 and RAB9A in the primary tumors from A375 cells.”

absence[Methods]

No reporting guideline mentioned.

WARNReviewer 2· 80% conf

Methods are detailed enough for replication, limitations are briefly discussed, and conclusions are proportional. However, no trial registration (N/A), no reporting guideline is referenced, and funding/COI is stated but no grant numbers. The data availability statement is contradictory.

Evidence

paraphrase[Methods, sections 2.1-2.18]

“Methods describe sample collection, cell culture, transfection, assays, and statistical analysis in sufficient detail.”

direct quote[Discussion, paragraph 3]

“However, the in vivo experiment only investigated the expression of miR-637 and RAB9A in the primary tumors from A375 cells.”

absence[Methods]

No reporting guideline is referenced.

WARNReviewer 3· 80% conf

Methods are reasonably complete for replication, but no trial registration, reporting guideline, or pre-registration is present; limitations are discussed only briefly; conclusions are generally proportional; funding and COI are stated.

Evidence

paraphrase[Methods sections 2.1-2.18]

“The methods extensively describe cell culture, transfection, qRT-PCR, Western blot, and xenograft protocols.”

absence[Entire paper]

No reporting guideline (e.g., ARRIVE) is referenced.

direct quote[Discussion, paragraph 2]

“However, the in vivo experiment only investigated the expression of miR-637 and RAB9A in the primary tumors from A375 cells. Further investigation should focus on the function of miR-637 and RAB9A on tumor formation in vivo.”

17 copyedit issues flagged: mostly grammar, typo, consistency.

Checked 28 references: 15 verified — 12 unresolved, 1 not checked.

Reproducibility

2 data/code links checked; 2 live.

Αlpha¹ Rigor Action ItemsFinal pass before journal submission

This is a post-publication audit. The published paper has significant robustness concerns: the statistical analysis is too incomplete for independent verification, and the data availability statement is demonstrably false. An informed reader should weigh these issues heavily, and a correction or independent re-analysis is warranted.

1.
HIGHdata code
Correct the Data Availability Statement: replace 'Data sharing is not applicable to this article as no new data were created or analyzed in this study' with an accurate statement that new data were generated, and deposit raw data (e.g., qRT-PCR Cq values, Western blot images, tumor measurements) in a public repository with a DOI.
The current statement is factually false and misrepresents the study; it must be corrected to allow readers to access and verify the data.
2.
HIGHstatistics
Report exact p-values (e.g., p = 0.023) instead of only '* p < 0.05' for all statistical comparisons throughout the Results and figure legends.
Threshold-only p-values prevent independent verification of statistical claims and are below standard reporting requirements.
3.
HIGHstatistics
Provide effect sizes (e.g., Cohen's d, Pearson's r) and confidence intervals for all group comparisons alongside p-values.
Effect sizes and CIs are essential for interpreting the magnitude and precision of reported effects, and their absence is a major reporting gap.
4.
HIGHrigor
Add a statement in the Methods (Statistical analysis or Study Design) describing randomization and blinding procedures for cell culture and animal experiments, or justify why they were not feasible.
Randomization and blinding are standard rigor elements to minimize bias; their omission weakens confidence in the results.
5.
HIGHrigor
Include a power analysis or sample-size justification for the in vitro and in vivo experiments in the Methods.
Without a power analysis, it is unclear whether the sample sizes are sufficient to detect biologically meaningful effects.
6.
HIGHreporting
Clarify whether the 'three repeats' of cell experiments are biological or technical replicates, and define the unit of replication (e.g., independent cell culture passages) in the Methods.
The distinction is critical for proper interpretation of statistical variability and reproducibility.
7.
HIGHrigor
Report cell line authentication (e.g., STR profiling) and mycoplasma testing results for all cell lines used (A375, A2058, HEM, HUVECs) in the Methods.
Unvalidated cell lines can produce irreproducible results; authentication and mycoplasma status are essential for confidence in cell-based experiments.
8.
HIGHreporting
Reference an appropriate reporting guideline (e.g., ARRIVE for animal studies) in the Methods and attach the completed checklist as supplementary material.
Following a reporting guideline improves completeness and reproducibility of the study description.
9.
HIGHother
Verify the 12 references that were not found in Crossref/OpenAlex and correct or replace them as needed (list: Malignant melanoma: skin cancer‐diagnosis, prevention, and treatment; Malignant melanoma: beyond the basics; Screening for reducing morbidity and mortality in malignant melanoma; Circular RNAs: identification, biogenesis and function; The potential role of miR‐29 in health and cancer diagnosis, prognosis, and therapy; miR‐146a‐5p: expression, regulation, and functions in cancer; Association of miR‐760 with cancer: an overview; MiR‐320a inhibits malignant phenotype of melanoma cells via targeting PBX3; miR‐509‐3p suppresses migration, invasion, and epithelial‐ mesenchymal transition in melanoma cells by targeting collagen triple helix repeat containing 1; Knockdown of circ_0084043 suppresses the development of human melanoma cells through miR‐429/tribbles homolog 2 axis and Wnt/beta‐catenin pathway; Knockdown of Rab9 suppresses the progression of gastric cancer through regulation of Akt signaling pathway; RAB9A plays an oncogenic role in human liver cancer cells).
References that cannot be located in standard registries may be fabricated; each must be confirmed or replaced.
10.
MEDIUMreporting
Report demographic details (age, sex, clinical stage) of the 52 melanoma tissue donors in Methods section 2.1.
Clinical cohort characterisation is essential for understanding the generalisability of the expression profiling results.
11.
MEDIUMrigor
Provide a justification for using only male mice in the xenograft study (e.g., a scientific rationale) in Methods section 2.17.
Using only one sex without justification limits the generalisability of the findings and omits an important biological variable.
12.
MEDIUMreporting
Show individual data points (e.g., dot plots or scatter plots) alongside bar charts for all cell-based and animal experiments.
Individual data points allow readers to assess data distribution, variability, and potential outliers that bar charts obscure.
13.
MEDIUMrigor
Add a brief section on statistical assumption verification: state whether normality and equal variance were tested (and by which method) for each test used, and what was done if assumptions were violated.
Without verifying assumptions, the validity of parametric tests (t-test, ANOVA) cannot be assessed.
14.
MEDIUMreporting
Correct the figure panel labelling error in Figure 3 where two panels are labelled 'E': renumber them sequentially (e.g., D, E, F, G...).
Duplicate panel labels cause confusion and suggest a documentation error that should be corrected.
15.
LOWcopyedit
Fix the grammar and typo issues flagged in the copyedit report: correct verb tense in Methods 2.1 ('transfer' to 'transferred'), fix awkward phrasing in Results 3.2 and Introduction paragraph 1, and standardise the statistical significance phrasing from 'conspicuous' to 'significant'.
These minor issues reduce readability and professionalism but do not affect scientific validity.

Full text (40,233 chars)Analyzed: Jul 6, 2026

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