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Rigor-Reviewed Manuscript

Antioxidant Properties and Gastroprotective Effects of 2-(Ethylthio)Benzohydrazones on Ethanol-Induced Acute Gastric Mucosal Lesions in Rats

Name: Kaimen Rigor review — Antioxidant Properties and Gastroprotective Effects of 2-(Ethylthio)Benzohydrazones on Ethanol-Induced Acute Gastric Mucosal Lesions in Rats
Item: Antioxidant Properties and Gastroprotective Effects of 2-(Ethylthio)Benzohydrazones on Ethanol-Induced Acute Gastric Mucosal Lesions in Rats
Rating: 59
Author: Alpha1

Nazarbahjat N, Kadir FA, Ariffin A, Abdulla MA, Abdullah Z, Yehye WA.

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Kaimen Rigor review — 1★ (Critical), 0 critical, 8 high issues for “Antioxidant Properties and Gastroprotective Effects of 2-(Ethylthio)Benzohydrazones on Ethanol-Induced Acute Gastric Mucosal Lesions in Rats”

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This rigor review was examined and confirmed by Alpha1 Editorial · August 5, 2026

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Antioxidant Properties and Gastroprotective Effects of 2-(Ethylthio)Benzohydrazones on Ethanol-Induced Acute Gastric Mucosal Lesions in Rats

Nazarbahjat N, Kadir FA, Ariffin A, Abdulla MA, Abdullah Z, Yehye WA.

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Journal

DOI

10.1371/journal.pone.0156022

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Kaimen Rigor reviewed

Rigor-Reviewed Manuscript

Antioxidant Properties and Gastroprotective Effects of 2-(Ethylthio)Benzohydrazones on Ethanol-Induced Acute Gastric Mucosal Lesions in Rats

Nazarbahjat N, Kadir FA, Ariffin A, Abdulla MA, Abdullah Z, Yehye WA.

Paper source

Antioxidant Properties and Gastroprotective Effects of 2-(Ethylthio)Benzohydrazones on Ethanol-Induced Acute Gastric Mucosal Lesions in Rats

Read full text (PMC)Journal

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Links back to this Kaimen Rigor review.

Alpha1 Editorial

This rigor review was examined and confirmed by Alpha1 Editorial · August 5, 2026

Kaimen Rigor review

Kaimen Rigor reviewed

1 / 5

Critical — serious problems across many checks

Interventional Study

Critical

High-priority

How this rating was calculated▸

Started at 5★ — deductions

IntegrityIntegrity concern ×5−2.5★
ReportingStudy design not met−0.5★
ReportingKey resources not met−0.5★
ReportingStatistical analysis not met−0.5★
ReportingData & code availability not met−0.5★
LinksDead data/code link−0.25★

Model-reported confidence: moderateReviewer agreement: moderateEngine: 7.15.0Source: full text

Not verified in this report

References were not verified against Crossref/OpenAlex.
Statistics were not checked: no recomputable values were found in this text — no test statistic reported with its degrees of freedom, no effect estimate printed with both a 95% CI and a p-value, and no percentage printed with both its count and its denominator.

Biggest issues

01Study design lacks key rigor safeguards
highDisclosurereviewer-assessedFull detail
The study design lacks many essential rigor elements: randomization method is unspecified, no blinding, no power analysis, no outlier handling, and no independent replication.
“The rats were divided randomly into 5 groups of 6 rats each.”
Methods
02Key resources not identified
highDisclosurereviewer-assessedFull detail
Only the organism source is adequately identified; antibodies, reagents, and software lack catalog numbers/versions, leading to inadequate identification of key resources.
“Both proteins were purchased from Santa Cruz Biotechnology, Inc., California, USA.”
Methods
03Statistical reporting inadequate
highDisclosurereviewer-assessedFull detail
The statistical analysis is inadequately reported: the post-hoc test is not named, assumptions are not verified, p-values are only given as thresholds, effect sizes are not reported with confidence intervals, and the software version is missing.
“The statistical significance of differences between groups was assessed using a one-way analysis of variance (ANOVA) followed by post hoc analysis by the prism statistical package.”
Statistical analysis
04Data and code not shared
highDisclosurereviewer-assessedFull detail
The data availability statement is vague (data within the paper) and no repository deposit, accession numbers, or code sharing is provided, making it inadequate.
“Data Availability All relevant data are within the paper and its Supporting Information files.”
Data availability
05Declared data/code link does not resolve
mediumAvailabilityrecomputedFull detail
Dead link — nothing to verify.
“http://umresearch.um.edu.my/mainpage.php?module=Maklumat&kategori=86&id=63&papar=1”

Findings by category

101ArithmeticNumerical inconsistencies 102OverreachOverstated conclusions · partly checked 103AuthenticityData authenticity concerns 404DisclosureReporting gaps 105AvailabilityBroken references and links 06Copyedit15 minor suggestions

This Kaimen Rigor review uses Kaimen 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's main strengths are a clear scientific premise, adequate ethical approvals, and fairly detailed methods. Major weaknesses are the lack of study design rigor (no blinding, no power analysis, incomplete randomization), inadequate statistical reporting (threshold p-values only, no effect sizes), and insufficient identification of key resources (antibodies, reagents, software).

This post-publication audit is based on three independent reviewer runs (all from the same model) and a copyedit pass. The study type is interventional animal study. No citations were flagged (0 checked). No statistical inconsistencies were found due to threshold-only p-values (coverage limited). The reproducibility check found 1 dead link (the paper's link to supporting information may be dead). Integrity concerns include implausibly small SDs in acute toxicity data and a non-monotonic pH response.

Numerical inconsistencies

1 finding · worst low

Values that contradict each other or are impossible for the stated sample: recomputed p-values and test statistics, GRIM/GRIMMER checks on summary numbers, percentages against their own counts, totals against their parts, and estimates against their own confidence intervals.

Internal contradictions in the reported numbersAssessed

Overstated conclusions

1 finding · worst low

Conclusions that reach past what the paper's own results support — including a significance claim that no longer holds when the statistic is recomputed, and efficacy resting on an unvalidated surrogate endpoint.

Conclusions only partially backed by the presented evidenceAssessed

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

Data authenticity concerns

1 finding · worst low

An adversarial read for patterns associated with data that may not be genuine: results that look too clean, implausibly large effects, duplicated data or images, and methods that do not match the results reported.

Data look implausibly cleanAssessed

5 integrity concerns flagged (0 high).

Reporting gaps

4 findings · worst high

Required detail the manuscript never states — study design, biological variables, ethics approval and consent, key resources, statistical reporting, data and code availability, and overall transparency.

Data and code not sharedAssessed
Key resources not identifiedAssessed
Statistical reporting inadequateAssessed
Study design lacks key rigor safeguardsAssessed

Eight Dimensions4 pass · 4 fail

The introduction cites multiple references on the role of ROS in gastric ulcer, the limitations of current PPI therapy, and the antioxidant properties of Schiff bases. The rationale for synthesizing new 2-(ethylthio)benzohydrazones and selecting the best antioxidant for in vivo testing is clearly stated. The paper also acknowledges the limitations of current treatments, thus addressing limitations of prior research.

Sub-criteria

prior work citedADEQUATE

premise rationaleADEQUATE

limitations addressedADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 95% conf

The paper clearly establishes the scientific foundation by citing prior work on ROS, gastric ulcers, and Schiff bases, and provides a logical rationale linking the premise to the study objectives.

Evidence

paraphrase[Introduction, paragraphs 1-3]

“Reactive oxygen species (ROS) and free radicals such as superoxide anion radical, hydroxyl radicals, and lipid peroxides are the injurious species known to cause many degenerative diseases []. ... Although the usage of proton-pump inhibitors (PPI) as a classic anti-ulcer drug therapy has revolutionized treatment of peptic ulcers and other gastrointestinal disorders, there is still no complete cure for this disease.”

“Thus, our strategy involves the synthesis of new Schiff bases, 2-(ethylthio)benzohydrazone derivatives. Among them, the most powerful antioxidant Schiff base, whether through hydrogen or electron transfer mechanisms, will be selected in order to illustrate its anti-ulcer effect on ethanol-induced gastric mucosal lesions in rats.”

Introduction

paraphrase[Introduction, paragraph 3]

“It has been shown that long term use of these drugs may be associated with ineffectiveness of different drug regimens and even resistance to these drugs are emerging.”

PASSReviewer 2· 95% conf

The paper cites prior research on ROS, gastric ulcer, and Schiff bases, provides a logical rationale, and addresses limitations of current therapies.

Evidence

“Reactive oxygen species (ROS) and free radicals such as superoxide anion radical, hydroxyl radicals, and lipid peroxides are the injurious species known to cause many degenerative diseases []. There are evidences for the aid of ROS in the etiology and pathophysiology of human diseases, such as neurodegenerative disorders, viral infections, autoimmune gastrointestinal inflammation and peptic ulcer [, ].”

Introduction

“It has been shown that long term use of these drugs may be associated with ineffectiveness of different drug regimens and even resistance to these drugs are emerging [].”

Introduction

PASSReviewer 3· 85% conf

The introduction establishes a clear rationale linking oxidative stress and gastric ulcer pathology to the rationale for testing new antioxidant Schiff-base derivatives, with prior work cited throughout.

Evidence

paraphrase[Introduction]

“Thus, our strategy involves the synthesis of new Schiff bases, 2-(ethylthio)benzohydrazone derivatives. Among them, the most powerful antioxidant Schiff base ... will be selected in order to illustrate its anti-ulcer effect on ethanol-induced gastric mucosal lesions in rats.”

paraphrase[Introduction]

“Although the usage of proton-pump inhibitors (PPI) as a classic anti-ulcer drug therapy has revolutionized treatment ... there is still no complete cure for this disease.”

The paper states that rats were 'divided randomly' but does not specify the randomization method (e.g., RNG, block). The unit of randomization is implicit (individual rat) but not explicitly stated. No blinding of investigators or assessors is reported. No a priori power analysis or sample size calculation is provided. Inclusion criteria are given (age, weight, health) but exclusion criteria are not mentioned. Outlier handling is not addressed. Positive and negative controls are present (ulcer control, omeprazole, normal). No independent replication of key findings is reported or planned. Only 1 of 9 applicable sub-criteria (controls) is adequate, resulting in a fail.

Sub-criteria

controlsADEQUATE

power analysisNOT REPORTED

blinding levelsNOT REPORTED

outlier handlingNOT REPORTED

randomization unitINADEQUATE

inclusion exclusionINADEQUATE

randomization methodINADEQUATE

replicate distinctionINADEQUATE

independent replicationNOT REPORTED

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 98% conf

The study design lacks many essential rigor elements: randomization method is unspecified, no blinding, no power analysis, no outlier handling, and no independent replication.

Evidence

“The rats were divided randomly into 5 groups of 6 rats each.”

Methods

absence[Materials and Methods]

No blinding is reported.

absence[Materials and Methods]

No power analysis is reported.

FAILReviewer 2· 95% conf

The study design lacks critical rigor elements: randomization method not specified, no blinding, no power analysis, no pre-specified exclusion criteria, no outlier handling, and no independent replication.

Evidence

“The rats were divided randomly into 5 groups of 6 rats each.”

Methods

absence[Entire paper]

No mention of blinding.

absence[Entire paper]

No power analysis or sample size calculation.

WARNReviewer 3· 80% conf

Adequate controls (positive omeprazole, ulcer control, normal group) and biological replicate definition (6 rats/group) are present, but randomization method, blinding, sample-size justification, and inclusion/exclusion criteria are not reported.

Evidence

“The rats were divided randomly into 5 groups of 6 rats each.”

Methods

“The reference group received oral doses of 20 mg/kg omeprazole in 10%Tween 20 (5 ml/kg) as positive control.”

Methods

absence[Materials and Methods]

No a priori power analysis or sample-size justification is given.

The paper reports that adult healthy female Sprague Dawley rats (6-8 weeks old, 180-225 g) were used, sourced from the Animal House, University of Malaya. Housing conditions are partially described: standard pellets and tap water, individual cages with wire-mesh bottoms. However, no justification is given for using only females, and housing details such as light cycle, temperature, and enrichment are missing. Three of five applicable sub-criteria are adequate (sex, age/weight/health, species/strain/source), meeting the 60% threshold for pass.

Sub-criteria

demographicsN/A

sex reportedADEQUATE

sex justifiedNOT REPORTED

age weight healthADEQUATE

housing conditionsINADEQUATE

species strain sourceADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 85% conf

Sex, age, weight, health status, species, strain, and source are reported, but sex is not justified and housing conditions lack details on light cycle and temperature.

Evidence

“Adult healthy female Sprague Dawley rats (6–8 weeks old) were obtained from the Animal House Experimental Unit, Faculty of Medicine, University of Malaya, Kuala Lumpur.”

Methods

absence[Materials and Methods]

No justification for using only females is provided.

paraphrase[Materials and Methods, Experimental animals for gastric ulcer]

“The rats were fed on standard pellets and tap water. Each rat was placed individually in a separate cage (one rat per cage) with wide-mesh wire bottoms to prevent coprophagia during the experiment.”

PASSReviewer 2· 85% conf

Sex, age, weight, health status, and species/strain/source are adequately reported, but housing conditions are only partially described and single-sex justification is missing.

Evidence

“Adult healthy female Sprague Dawley rats (6–8 weeks old) were obtained from the Animal House Experimental Unit, Faculty of Medicine, University of Malaya, Kuala Lumpur. The rats weighed between 180–200 g.”

Methods

“Each rat was placed individually in a separate cage (one rat per cage) with wide-mesh wire bottoms to prevent coprophagia during the experiment. The animals were fed on standard pellets and tap water.”

Methods

absence[Entire paper]

No justification for single-sex study.

PASSReviewer 3· 80% conf

Sex, age, weight, strain, source, and housing are reported for the rats; the only gap is lack of scientific justification for using a single sex (females).

Evidence

Methods

“Each rat was placed individually in a separate cage (one rat per cage) with wide-mesh wire bottoms to prevent coprophagia during the experiment.”

Methods

absence[Materials and Methods]

No rationale given for using only female rats.

The ethics statement explicitly names the Institutional Animal Care and Use Committee of University of Malaya (UM IACUC) and provides two ethic numbers. It also states that the study was carried out according to the criteria outlined in the 'Guide for the Care and Use of Laboratory Animals.' Both applicable sub-criteria (iacuc_statement, regulatory_compliance) are reported and adequate.

Sub-criteria

iacuc statementADEQUATE

informed consentN/A

irb ethics statementN/A

regulatory complianceADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 98% conf

The study has a named IACUC approval with protocol numbers and states compliance with the Guide for the Care and Use of Laboratory Animals.

Evidence

“The protocol was approved by the Institutional Animal Care and Use Committee of University of Malaya (UM IACUC) with Ethic Numbers: PM/07/05/2012/MAA (a) (R) and PM/28/09/2012/MAA (R).”

Methods

paraphrase[Materials and Methods, Ethics statement]

“This study was carried out according to the criteria outlined in the "Guide for the Care and Use of laboratory Animals" prepared by the National Academy of Sciences and published by the national Institute of health.”

PASSReviewer 2· 95% conf

Animal ethics approval from a named committee with protocol numbers is provided, and regulatory compliance with the Guide for the Care and Use of Laboratory Animals is stated.

Evidence

“The protocol was approved by the Institutional Animal Care and Use Committee of University of Malaya (UM IACUC) with Ethic Numbers: PM/07/05/2012/MAA (a) (R) and PM/28/09/2012/MAA (R).”

Methods

paraphrase[Materials and Methods, Ethics statement]

“This study was carried out according to the criteria outlined in the 'Guide for the Care and Use of laboratory Animals' prepared by the National Academy of Sciences and published by the national Institute of health.”

PASSReviewer 3· 90% conf

The animal study has a named IACUC approval with protocol numbers and a named regulatory framework (Guide for the Care and Use of Laboratory Animals).

Evidence

“The protocol was approved by the Institutional Animal Care and Use Committee of University of Malaya (UM IACUC) with Ethic Numbers: PM/07/05/2012/MAA (a) (R) and PM/28/09/2012/MAA (R).”

Methods

paraphrase[Materials and Methods, Ethics statement]

Antibodies (HSP70 and Bax) are identified by vendor only, with no catalog number, clone, or validation data. Reagents (chemicals, omeprazole) are named with source but no catalog numbers. Statistical software 'prism' is mentioned without version. The organism (Sprague Dawley rats) is adequately identified with source.

Sub-criteria

mycoplasma testingN/A

reagents identifiedINADEQUATE

organisms identifiedADEQUATE

antibodies identifiedINADEQUATE

cell line authenticationN/A

software tools identifiedINADEQUATE

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 90% conf

Key resources are inadequately identified: antibodies lack catalog numbers and dilutions, reagents like omeprazole lack lot numbers, and software lacks version information.

Evidence

“Both proteins were purchased from Santa Cruz Biotechnology, Inc., California, USA.”

Methods

“In this study, omeprazole was used as the reference anti-ulcer drug, and was obtained from the University Malaya Medical Centre Pharmacy.”

Methods

“The statistical significance of differences between groups was assessed using a one-way analysis of variance (ANOVA) followed by post hoc analysis by the prism statistical package.”

Statistical analysis

FAILReviewer 2· 95% conf

Only the organism source is adequately identified; antibodies, reagents, and software lack catalog numbers/versions, leading to inadequate identification of key resources.

Evidence

“Both proteins were purchased from Santa Cruz Biotechnology, Inc., California, USA.”

Methods

“All chemicals and solvents were of analytical grade and purchased from Aldrich and Merck.”

Methods

“The statistical significance of differences between groups was assessed using a one-way analysis of variance (ANOVA) followed by post hoc analysis by the prism statistical package.”

Methods

WARNReviewer 3· 75% conf

Organisms and reagents are identified, but antibodies (HSP-70, Bax) lack catalog numbers/RRIDs/validation details and the statistical software has no version specified.

Evidence

“Both proteins were purchased from Santa Cruz Biotechnology, Inc., California, USA.”

Methods

“assessed using a one-way analysis of variance (ANOVA) followed by post hoc analysis by the prism statistical package.”

Methods

The paper states that ANOVA followed by post hoc analysis was used, but does not name the post-hoc test (e.g., Tukey, Dunnett). No verification of test assumptions (normality, equal variance) is reported. P-values are reported only as 'p<0.05' without exact values. Effect sizes are given as mean ± SEM, but no confidence intervals for effect sizes or differences are provided. The software (prism) is named but version is not given. Data presentation includes per-group n and SEM, but no individual data points are shown. Mathematical plausibility checks revealed no arithmetic errors. Only 1 of 7 applicable sub-criteria (mathematical_plausibility) is adequate, resulting in a fail.

Sub-criteria

tests namedINADEQUATE

exact p valuesINADEQUATE

effect sizes ciNOT REPORTED

data presentationINADEQUATE

software identifiedINADEQUATE

assumptions verifiedNOT REPORTED

mathematical plausibilityPASS

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 95% conf

The statistical analysis is inadequately reported: the post-hoc test is not named, assumptions are not verified, p-values are only given as thresholds, effect sizes are not reported with confidence intervals, and the software version is missing.

Evidence

“The statistical significance of differences between groups was assessed using a one-way analysis of variance (ANOVA) followed by post hoc analysis by the prism statistical package.”

Statistical analysis

absence[Statistical analysis]

No verification of assumptions reported.

“A value of p<0.05 was considered significant.”

Results

FAILReviewer 2· 95% conf

Only the test name is adequately reported; assumptions are not verified, p-values are reported only as thresholds, effect sizes lack confidence intervals, software version is missing, and data presentation does not show individual points.

Evidence

“The statistical significance of differences between groups was assessed using a one-way analysis of variance (ANOVA) followed by post hoc analysis by the prism statistical package.”

Methods

“Values are assumed as mean ± S.E.M. The statistical analysis was assessed with one-way ANOVA (post hoc analysis) with P < 0.05.”

Table 4

absence[Results, Table 4]

No confidence intervals for effect sizes.

WARNReviewer 3· 75% conf

ANOVA with post-hoc analysis is named and per-group data are presented as mean±SEM, but only threshold p-values ('p<0.05') are reported, effect sizes/CIs are absent, and assumptions are not verified.

Evidence

“The statistical significance of differences between groups was assessed using a one-way analysis of variance (ANOVA) followed by post hoc analysis by the prism statistical package.”

Methods

“The statistical analysis was assessed with one-way ANOVA (post hoc analysis) with P < 0.05.”

Table 4

paraphrase[Results, Table 4]

“Inhibition values (79%, 92.6%, 97.8%) match (850-treat)/850 for the tabulated ulcer-area means.”

The data availability statement says 'All relevant data are within the paper and its Supporting Information files', which is not a concrete route to a repository. No raw data are deposited in a public repository with a persistent identifier. No accession numbers or custom code are provided. Supporting information files are part of the journal article but not a separate repository deposit.

Sub-criteria

code sharingN/A

accession numbersNOT REPORTED

repository depositNOT REPORTED

data availability statementINADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 90% conf

The data availability statement indicates that all relevant data are within the paper and its Supporting Information files, which is adequate for this study; no repository or code is required.

Evidence

“All relevant data are within the paper and its Supporting Information files.”

Data availability

FAILReviewer 2· 95% conf

The data availability statement is vague (data within the paper) and no repository deposit, accession numbers, or code sharing is provided, making it inadequate.

Evidence

“Data Availability All relevant data are within the paper and its Supporting Information files.”

Data availability

absence[Entire paper]

No repository deposit.

absence[Entire paper]

No code sharing.

FAILReviewer 3· 70% conf

The only data-availability statement is a vague 'all relevant data are within the paper and its Supporting Information files' with no repository, accession, or concrete access mechanism.

Evidence

“Data Availability All relevant data are within the paper and its Supporting Information files.”

Data availability

absence[Data Availability]

No raw data deposited in a public repository.

The methods section provides sufficient detail for synthesis and in vivo experiments, including protocols for DPPH, FRAP, ulcer induction, histology, and IHC. All outcomes mentioned in methods are reported in results. Conclusions are proportional to the evidence, with no overclaiming of causality. Funding sources and a competing interests statement are provided. However, the paper does not reference the ARRIVE guidelines for animal studies, and there is no dedicated limitations section. Four of six applicable sub-criteria are adequate, meeting the 60% threshold for pass.

Sub-criteria

funding coiADEQUATE

trial registrationN/A

reporting guidelineNOT REPORTED

methods completenessADEQUATE

all outcomes reportedADEQUATE

limitations discussedNOT REPORTED

conclusions proportionalADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 85% conf

The methods are detailed enough for replication, all outcomes are reported, conclusions are proportional, and funding/COI are disclosed, but the ARRIVE reporting guideline is not referenced and limitations are not discussed.

Evidence

paraphrase[Materials and Methods, entire section]

“General procedure for the synthesis of 2-(ethylsulfanyl)-N'-[(substitutedphenyl)methylidene]benzohydrazide (1–6) ... To a warm stirred solution of aryl aldehyde (3 mmol in 20 ml absolute ethanol), 2-(ethylsulfanyl)benzo hydrazide (3 mmol) was added in small portion, and refluxed for 7 hours, after cooling the mixture, a precipitate was washed with cold ethanol, then recrystallized from suitable solvent.”

absence[Materials and Methods]

No reference to ARRIVE guidelines.

“Competing Interests: The authors have declared that no competing interests exist.”

Funding

PASSReviewer 2· 85% conf

Methods are complete, all outcomes are reported, conclusions are proportional, and funding/COI is disclosed, but limitations are not discussed and no reporting guideline is referenced.

Evidence

paraphrase[Materials and Methods]

“The paper describes synthesis, DPPH and FRAP assays, animal protocol, histology, and immunohistochemistry in sufficient detail.”

“The possible mechanisms underlying this gastroprotective effect of compound 1 is due to potent antioxidant activity besides HSP-70 up regulation and Bax protein down regulation.”

Conclusion

“Competing Interests: The authors have declared that no competing interests exist.”

Funding

PASSReviewer 3· 75% conf

Methods are detailed, funding and COI are disclosed, and conclusions are appropriately hedged; the main gaps are no ARRIVE reporting guideline and limited explicit limitations discussion.

Evidence

“Competing Interests: The authors have declared that no competing interests exist.”

Funding

paraphrase[Discussion]

“The discussion cites prior studies and mechanisms but does not explicitly enumerate this study's limitations or biases.”

absence[Entire paper]

No ARRIVE or other reporting guideline referenced.

Broken references and links

1 finding · worst medium

References checked against Crossref, OpenAlex and Retraction Watch for retractions and resolvability, plus declared data and code links probed for whether they resolve to content matching the paper.

Dead data/code linksRecomputed

1 data/code link checked; 0 live, 1 dead.

Copyediting

15 minor

Wording, consistency and formatting errors that need correcting before submission.

No major wording or formatting errors. 15 minor suggestions below.

15 copyedit issues flagged: mostly typo, clarity, punctuation.

Kaimen Rigor Action ItemsFinal pass before journal submission

In this post-publication audit, the paper's robustness is limited by missing rigor elements (no blinding, no power analysis, no exact p-values, incomplete resource identification). An informed reader should weigh these gaps heavily; the claims of gastroprotection and the statistical significance are weakened by the absence of standard reporting practices. A correction or independent re-analysis would be warranted to address the statistical reporting and the missing blinding.

1.
HIGHrigor
Acknowledge the lack of blinding, unspecified randomization method, and absence of a power analysis in a correction; discuss the impact on the reliability of the results.
These are fundamental design gaps that undermine the internal validity of the study.
2.
HIGHstatistics
Provide exact p-values and confidence intervals for all reported comparisons in a correction or re-analysis.
Threshold-only p-values (p<0.05) are insufficient for interpretation and reproducibility.
3.
HIGHdata code
Disclose catalog numbers, clone/RRID, and dilution for the HSP-70 and Bax antibodies in a correction.
Antibodies are key resources; missing identifiers impair reproducibility of the immunohistochemistry results.
4.
HIGHother
Clarify or correct the acute toxicity data in Table 3, particularly the implausibly small standard deviations (TB=2.00±0.006, vehicle=2.00±0.00).
Such small SDs are suspicious and may indicate a data reporting error or fabrication.
5.
HIGHrigor
State the version of the statistical software (e.g., GraphPad Prism 5.0) in the Statistical analysis section.
Software version is necessary for reproducibility of the analysis.
6.
HIGHreporting
Add a dedicated limitations section to the Discussion, addressing the single-sex design, small sample size, lack of blinding, and potential confounding factors.
Transparency about limitations helps readers interpret the study's scope.
7.
HIGHreporting
Reference the ARRIVE guidelines for animal research in the Methods or Ethics statement.
Adherence to reporting guidelines is expected for preclinical animal studies.
8.
HIGHdata code
Deposit raw data (e.g., animal measurements, histology scores) in a public repository (e.g., Figshare, Dryad) and update the Data Availability statement with a DOI.
A vague statement ('data within the paper') does not provide a concrete access route; a repository deposit ensures long-term accessibility.
9.
MEDIUMcopyedit
Reconcile the inconsistent use of SD vs. SEM across tables (Table 1 footnote uses SD, others use SEM).
Consistency in reporting dispersion is essential for clarity.
10.
MEDIUMcopyedit
Correct the typo '10% Twee20' to '10% Tween 20' throughout the manuscript.
Typographical errors can undermine confidence in the reporting.
11.
MEDIUMcopyedit
In the DPPH section, replace 'minimum inhibitory concentration IC50' with 'IC50' to avoid confusion with antimicrobial terminology.
IC50 in antioxidant assays is not a minimum inhibitory concentration; accurate terminology is important.
12.
LOWcopyedit
Correct the author name formatting in the Abstract (e.g., 'Nazarbahjat Nafal' may need to be reversed).
Author name formatting should be consistent with journal requirements.

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Antioxidant Properties and Gastroprotective Effects of 2-(Ethylthio)Benzohydrazones on Ethanol-Induced Acute Gastric Mucosal Lesions in Rats

Nazarbahjat N, Kadir FA, Ariffin A, Abdulla MA, Abdullah Z, Yehye WA.

Read full text (PMC)

Journal

DOI

10.1371/journal.pone.0156022

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Issued Aug 5, 2026Engine 7.15.0

Alpha1 assesses methodological rigor, not the importance of the findings. See how we calibrate →

Kaimen Rigor review

Kaimen Rigor reviewed

1 / 5

Critical — serious problems across many checks

Interventional Study

Critical

High-priority

How this rating was calculated▸

Started at 5★ — deductions

IntegrityIntegrity concern ×5−2.5★
ReportingStudy design not met−0.5★
ReportingKey resources not met−0.5★
ReportingStatistical analysis not met−0.5★
ReportingData & code availability not met−0.5★
LinksDead data/code link−0.25★

Model-reported confidence: moderateReviewer agreement: moderateEngine: 7.15.0Source: full text

Not verified in this report

References were not verified against Crossref/OpenAlex.
Statistics were not checked: no recomputable values were found in this text — no test statistic reported with its degrees of freedom, no effect estimate printed with both a 95% CI and a p-value, and no percentage printed with both its count and its denominator.

Biggest issues

01Study design lacks key rigor safeguards
highDisclosurereviewer-assessedFull detail
The study design lacks many essential rigor elements: randomization method is unspecified, no blinding, no power analysis, no outlier handling, and no independent replication.
“The rats were divided randomly into 5 groups of 6 rats each.”
Methods
02Key resources not identified
highDisclosurereviewer-assessedFull detail
Only the organism source is adequately identified; antibodies, reagents, and software lack catalog numbers/versions, leading to inadequate identification of key resources.
“Both proteins were purchased from Santa Cruz Biotechnology, Inc., California, USA.”
Methods
03Statistical reporting inadequate
highDisclosurereviewer-assessedFull detail
The statistical analysis is inadequately reported: the post-hoc test is not named, assumptions are not verified, p-values are only given as thresholds, effect sizes are not reported with confidence intervals, and the software version is missing.
“The statistical significance of differences between groups was assessed using a one-way analysis of variance (ANOVA) followed by post hoc analysis by the prism statistical package.”
Statistical analysis
04Data and code not shared
highDisclosurereviewer-assessedFull detail
The data availability statement is vague (data within the paper) and no repository deposit, accession numbers, or code sharing is provided, making it inadequate.
“Data Availability All relevant data are within the paper and its Supporting Information files.”
Data availability
05Declared data/code link does not resolve
mediumAvailabilityrecomputedFull detail
Dead link — nothing to verify.
“http://umresearch.um.edu.my/mainpage.php?module=Maklumat&kategori=86&id=63&papar=1”

Findings by category

Numerical inconsistencies

1 finding · worst low

Internal contradictions in the reported numbersAssessed

Overstated conclusions

1 finding · worst low

Conclusions only partially backed by the presented evidenceAssessed

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

Data authenticity concerns

1 finding · worst low

Data look implausibly cleanAssessed

5 integrity concerns flagged (0 high).

Reporting gaps

4 findings · worst high

Data and code not sharedAssessed
Key resources not identifiedAssessed
Statistical reporting inadequateAssessed
Study design lacks key rigor safeguardsAssessed

Eight Dimensions4 pass · 4 fail

Sub-criteria

prior work citedADEQUATE

premise rationaleADEQUATE

limitations addressedADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 95% conf

The paper clearly establishes the scientific foundation by citing prior work on ROS, gastric ulcers, and Schiff bases, and provides a logical rationale linking the premise to the study objectives.

Evidence

paraphrase[Introduction, paragraphs 1-3]

Introduction

paraphrase[Introduction, paragraph 3]

“It has been shown that long term use of these drugs may be associated with ineffectiveness of different drug regimens and even resistance to these drugs are emerging.”

PASSReviewer 2· 95% conf

The paper cites prior research on ROS, gastric ulcer, and Schiff bases, provides a logical rationale, and addresses limitations of current therapies.

Evidence

“Reactive oxygen species (ROS) and free radicals such as superoxide anion radical, hydroxyl radicals, and lipid peroxides are the injurious species known to cause many degenerative diseases []. There are evidences for the aid of ROS in the etiology and pathophysiology of human diseases, such as neurodegenerative disorders, viral infections, autoimmune gastrointestinal inflammation and peptic ulcer [, ].”

Introduction

“It has been shown that long term use of these drugs may be associated with ineffectiveness of different drug regimens and even resistance to these drugs are emerging [].”

Introduction

PASSReviewer 3· 85% conf

Evidence

paraphrase[Introduction]

“Although the usage of proton-pump inhibitors (PPI) as a classic anti-ulcer drug therapy has revolutionized treatment ... there is still no complete cure for this disease.”

Sub-criteria

controlsADEQUATE

power analysisNOT REPORTED

blinding levelsNOT REPORTED

outlier handlingNOT REPORTED

randomization unitINADEQUATE

inclusion exclusionINADEQUATE

randomization methodINADEQUATE

replicate distinctionINADEQUATE

independent replicationNOT REPORTED

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 98% conf

The study design lacks many essential rigor elements: randomization method is unspecified, no blinding, no power analysis, no outlier handling, and no independent replication.

Evidence

“The rats were divided randomly into 5 groups of 6 rats each.”

Methods

absence[Materials and Methods]

No blinding is reported.

absence[Materials and Methods]

No power analysis is reported.

FAILReviewer 2· 95% conf

Evidence

“The rats were divided randomly into 5 groups of 6 rats each.”

Methods

absence[Entire paper]

No mention of blinding.

absence[Entire paper]

No power analysis or sample size calculation.

WARNReviewer 3· 80% conf

Evidence

“The rats were divided randomly into 5 groups of 6 rats each.”

Methods

“The reference group received oral doses of 20 mg/kg omeprazole in 10%Tween 20 (5 ml/kg) as positive control.”

Methods

absence[Materials and Methods]

No a priori power analysis or sample-size justification is given.

Sub-criteria

demographicsN/A

sex reportedADEQUATE

sex justifiedNOT REPORTED

age weight healthADEQUATE

housing conditionsINADEQUATE

species strain sourceADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 85% conf

Sex, age, weight, health status, species, strain, and source are reported, but sex is not justified and housing conditions lack details on light cycle and temperature.

Evidence

“Adult healthy female Sprague Dawley rats (6–8 weeks old) were obtained from the Animal House Experimental Unit, Faculty of Medicine, University of Malaya, Kuala Lumpur.”

Methods

absence[Materials and Methods]

No justification for using only females is provided.

paraphrase[Materials and Methods, Experimental animals for gastric ulcer]

PASSReviewer 2· 85% conf

Sex, age, weight, health status, and species/strain/source are adequately reported, but housing conditions are only partially described and single-sex justification is missing.

Evidence

Methods

absence[Entire paper]

No justification for single-sex study.

PASSReviewer 3· 80% conf

Sex, age, weight, strain, source, and housing are reported for the rats; the only gap is lack of scientific justification for using a single sex (females).

Evidence

Methods

“Each rat was placed individually in a separate cage (one rat per cage) with wide-mesh wire bottoms to prevent coprophagia during the experiment.”

Methods

absence[Materials and Methods]

No rationale given for using only female rats.

Sub-criteria

iacuc statementADEQUATE

informed consentN/A

irb ethics statementN/A

regulatory complianceADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 98% conf

The study has a named IACUC approval with protocol numbers and states compliance with the Guide for the Care and Use of Laboratory Animals.

Evidence

“The protocol was approved by the Institutional Animal Care and Use Committee of University of Malaya (UM IACUC) with Ethic Numbers: PM/07/05/2012/MAA (a) (R) and PM/28/09/2012/MAA (R).”

Methods

paraphrase[Materials and Methods, Ethics statement]

PASSReviewer 2· 95% conf

Animal ethics approval from a named committee with protocol numbers is provided, and regulatory compliance with the Guide for the Care and Use of Laboratory Animals is stated.

Evidence

“The protocol was approved by the Institutional Animal Care and Use Committee of University of Malaya (UM IACUC) with Ethic Numbers: PM/07/05/2012/MAA (a) (R) and PM/28/09/2012/MAA (R).”

Methods

paraphrase[Materials and Methods, Ethics statement]

PASSReviewer 3· 90% conf

The animal study has a named IACUC approval with protocol numbers and a named regulatory framework (Guide for the Care and Use of Laboratory Animals).

Evidence

“The protocol was approved by the Institutional Animal Care and Use Committee of University of Malaya (UM IACUC) with Ethic Numbers: PM/07/05/2012/MAA (a) (R) and PM/28/09/2012/MAA (R).”

Methods

paraphrase[Materials and Methods, Ethics statement]

Sub-criteria

mycoplasma testingN/A

reagents identifiedINADEQUATE

organisms identifiedADEQUATE

antibodies identifiedINADEQUATE

cell line authenticationN/A

software tools identifiedINADEQUATE

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 90% conf

Key resources are inadequately identified: antibodies lack catalog numbers and dilutions, reagents like omeprazole lack lot numbers, and software lacks version information.

Evidence

“Both proteins were purchased from Santa Cruz Biotechnology, Inc., California, USA.”

Methods

“In this study, omeprazole was used as the reference anti-ulcer drug, and was obtained from the University Malaya Medical Centre Pharmacy.”

Methods

“The statistical significance of differences between groups was assessed using a one-way analysis of variance (ANOVA) followed by post hoc analysis by the prism statistical package.”

Statistical analysis

FAILReviewer 2· 95% conf

Only the organism source is adequately identified; antibodies, reagents, and software lack catalog numbers/versions, leading to inadequate identification of key resources.

Evidence

“Both proteins were purchased from Santa Cruz Biotechnology, Inc., California, USA.”

Methods

“All chemicals and solvents were of analytical grade and purchased from Aldrich and Merck.”

Methods

“The statistical significance of differences between groups was assessed using a one-way analysis of variance (ANOVA) followed by post hoc analysis by the prism statistical package.”

Methods

WARNReviewer 3· 75% conf

Organisms and reagents are identified, but antibodies (HSP-70, Bax) lack catalog numbers/RRIDs/validation details and the statistical software has no version specified.

Evidence

“Both proteins were purchased from Santa Cruz Biotechnology, Inc., California, USA.”

Methods

“assessed using a one-way analysis of variance (ANOVA) followed by post hoc analysis by the prism statistical package.”

Methods

Sub-criteria

tests namedINADEQUATE

exact p valuesINADEQUATE

effect sizes ciNOT REPORTED

data presentationINADEQUATE

software identifiedINADEQUATE

assumptions verifiedNOT REPORTED

mathematical plausibilityPASS

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 95% conf

Evidence

“The statistical significance of differences between groups was assessed using a one-way analysis of variance (ANOVA) followed by post hoc analysis by the prism statistical package.”

Statistical analysis

absence[Statistical analysis]

No verification of assumptions reported.

“A value of p<0.05 was considered significant.”

Results

FAILReviewer 2· 95% conf

Evidence

“The statistical significance of differences between groups was assessed using a one-way analysis of variance (ANOVA) followed by post hoc analysis by the prism statistical package.”

Methods

“Values are assumed as mean ± S.E.M. The statistical analysis was assessed with one-way ANOVA (post hoc analysis) with P < 0.05.”

Table 4

absence[Results, Table 4]

No confidence intervals for effect sizes.

WARNReviewer 3· 75% conf

Evidence

“The statistical significance of differences between groups was assessed using a one-way analysis of variance (ANOVA) followed by post hoc analysis by the prism statistical package.”

Methods

“The statistical analysis was assessed with one-way ANOVA (post hoc analysis) with P < 0.05.”

Table 4

paraphrase[Results, Table 4]

“Inhibition values (79%, 92.6%, 97.8%) match (850-treat)/850 for the tabulated ulcer-area means.”

Sub-criteria

code sharingN/A

accession numbersNOT REPORTED

repository depositNOT REPORTED

data availability statementINADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 90% conf

The data availability statement indicates that all relevant data are within the paper and its Supporting Information files, which is adequate for this study; no repository or code is required.

Evidence

“All relevant data are within the paper and its Supporting Information files.”

Data availability

FAILReviewer 2· 95% conf

The data availability statement is vague (data within the paper) and no repository deposit, accession numbers, or code sharing is provided, making it inadequate.

Evidence

“Data Availability All relevant data are within the paper and its Supporting Information files.”

Data availability

absence[Entire paper]

No repository deposit.

absence[Entire paper]

No code sharing.

FAILReviewer 3· 70% conf

The only data-availability statement is a vague 'all relevant data are within the paper and its Supporting Information files' with no repository, accession, or concrete access mechanism.

Evidence

“Data Availability All relevant data are within the paper and its Supporting Information files.”

Data availability

absence[Data Availability]

No raw data deposited in a public repository.

Sub-criteria

funding coiADEQUATE

trial registrationN/A

reporting guidelineNOT REPORTED

methods completenessADEQUATE

all outcomes reportedADEQUATE

limitations discussedNOT REPORTED

conclusions proportionalADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 85% conf

Evidence

paraphrase[Materials and Methods, entire section]

absence[Materials and Methods]

No reference to ARRIVE guidelines.

“Competing Interests: The authors have declared that no competing interests exist.”

Funding

PASSReviewer 2· 85% conf

Methods are complete, all outcomes are reported, conclusions are proportional, and funding/COI is disclosed, but limitations are not discussed and no reporting guideline is referenced.

Evidence

paraphrase[Materials and Methods]

“The paper describes synthesis, DPPH and FRAP assays, animal protocol, histology, and immunohistochemistry in sufficient detail.”

“The possible mechanisms underlying this gastroprotective effect of compound 1 is due to potent antioxidant activity besides HSP-70 up regulation and Bax protein down regulation.”

Conclusion

“Competing Interests: The authors have declared that no competing interests exist.”

Funding

PASSReviewer 3· 75% conf

Methods are detailed, funding and COI are disclosed, and conclusions are appropriately hedged; the main gaps are no ARRIVE reporting guideline and limited explicit limitations discussion.

Evidence

“Competing Interests: The authors have declared that no competing interests exist.”

Funding

paraphrase[Discussion]

“The discussion cites prior studies and mechanisms but does not explicitly enumerate this study's limitations or biases.”

absence[Entire paper]

No ARRIVE or other reporting guideline referenced.

Broken references and links

1 finding · worst medium

References checked against Crossref, OpenAlex and Retraction Watch for retractions and resolvability, plus declared data and code links probed for whether they resolve to content matching the paper.

Dead data/code linksRecomputed

1 data/code link checked; 0 live, 1 dead.

Copyediting

15 minor

Wording, consistency and formatting errors that need correcting before submission.

No major wording or formatting errors. 15 minor suggestions below.

15 copyedit issues flagged: mostly typo, clarity, punctuation.

Kaimen Rigor Action ItemsFinal pass before journal submission

1.
HIGHrigor
Acknowledge the lack of blinding, unspecified randomization method, and absence of a power analysis in a correction; discuss the impact on the reliability of the results.
These are fundamental design gaps that undermine the internal validity of the study.
2.
HIGHstatistics
Provide exact p-values and confidence intervals for all reported comparisons in a correction or re-analysis.
Threshold-only p-values (p<0.05) are insufficient for interpretation and reproducibility.
3.
HIGHdata code
Disclose catalog numbers, clone/RRID, and dilution for the HSP-70 and Bax antibodies in a correction.
Antibodies are key resources; missing identifiers impair reproducibility of the immunohistochemistry results.
4.
HIGHother
Clarify or correct the acute toxicity data in Table 3, particularly the implausibly small standard deviations (TB=2.00±0.006, vehicle=2.00±0.00).
Such small SDs are suspicious and may indicate a data reporting error or fabrication.
5.
HIGHrigor
State the version of the statistical software (e.g., GraphPad Prism 5.0) in the Statistical analysis section.
Software version is necessary for reproducibility of the analysis.
6.
HIGHreporting
Add a dedicated limitations section to the Discussion, addressing the single-sex design, small sample size, lack of blinding, and potential confounding factors.
Transparency about limitations helps readers interpret the study's scope.
7.
HIGHreporting
Reference the ARRIVE guidelines for animal research in the Methods or Ethics statement.
Adherence to reporting guidelines is expected for preclinical animal studies.
8.
HIGHdata code
Deposit raw data (e.g., animal measurements, histology scores) in a public repository (e.g., Figshare, Dryad) and update the Data Availability statement with a DOI.
A vague statement ('data within the paper') does not provide a concrete access route; a repository deposit ensures long-term accessibility.
9.
MEDIUMcopyedit
Reconcile the inconsistent use of SD vs. SEM across tables (Table 1 footnote uses SD, others use SEM).
Consistency in reporting dispersion is essential for clarity.
10.
MEDIUMcopyedit
Correct the typo '10% Twee20' to '10% Tween 20' throughout the manuscript.
Typographical errors can undermine confidence in the reporting.
11.
MEDIUMcopyedit
In the DPPH section, replace 'minimum inhibitory concentration IC50' with 'IC50' to avoid confusion with antimicrobial terminology.
IC50 in antioxidant assays is not a minimum inhibitory concentration; accurate terminology is important.
12.
LOWcopyedit
Correct the author name formatting in the Abstract (e.g., 'Nazarbahjat Nafal' may need to be reversed).
Author name formatting should be consistent with journal requirements.

Full text (50,790 chars)Analyzed: Aug 5, 2026