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.
- DOI
- 10.1371/journal.pone.0156022
- Record issued
- 2026-08-05
- Engine
- 7.15.0
- Exported
- 2026-09-19
Prepared by Alpha1. This document is confidential: it is intended for the recipient it was shared with and must not be redistributed. The live record at alpha1science.com/verify/a252731a-cc3e-44c6-bdf2-427ed73a1400 is authoritative.
How this rating was calculated
- 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★
- 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.
- 01Study design lacks key rigor safeguards
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
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
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
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
Dead link — nothing to verify.
“http://umresearch.um.edu.my/mainpage.php?module=Maklumat&kategori=86&id=63&papar=1”
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 lowValues 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
- lowinternal contradictionThe gastric pH values are non-monotonic across doses: compound 1 at 50 mg/kg gives pH 5.02 while 100 mg/kg gives pH 3.9 (more acidic), which is inconsistent with a dose-dependent antisecretory effect claimed in the text.
Compound 1 (50 mg/kg) | 5.02 ± 0.48 a | Compound 1 (100 mg/kg) | 3.9±0.9 a
Table 4reviewer’s wording - lowinternal contradictionThe paper uses SEM for in vivo data but SD for in vitro data without explanation, and the statistical analysis section states 'mean ± S.E.M.' while Table 1 footnote states 'mean ± SD'.
All values were reported as mean ± S.E.M. (Statistical analysis) vs. a Each value represents mean ± SD (Table 1 footnote)
Table 1reviewer’s wording - lowinternal contradictionThe acute toxicity study does not report the number of animals used per group, making it impossible to verify the claims of no toxicity.
The acute toxic study was used to determine a safe dose for synthesized compound. ... The rats were fasted overnight and next day the compound ... was administered orally at a dose level 500 mg/kg.
Methodsreviewer’s wording
Overstated conclusions
1 finding · worst lowConclusions 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.
- partialReviewer 1The 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.The paper shows antioxidant activity and IHC changes (HSP70 up, Bax down), but does not provide causal evidence linking these mechanisms to gastroprotection; it is a correlative observation.Evidence: In vitro antioxidant data (Table 1) and IHC images (Figures 6, 7) show HSP70 upregulation and Bax downregulation in compound 1-treated groups.
“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 - partialReviewer 3The gastroprotective effect is attributable to the antioxidant activity of compound 1.Compound 1 has in vitro antioxidant activity (DPPH IC50 0.69±0.08 μg/ml) and the paper hedges the causal link, but no in vivo oxidative-stress markers were measured to tie the antioxidant activity to the gastroprotection.Evidence: Table 1 DPPH/FRAP data for compound 1; Discussion speculation.
“In conclusion, the gastroprotective effect of compound 1 may be due to its antioxidant activity, and/or due to up-regulation of HSP70 and down-regulation of Bax protein in stained tissue section.”
Discussion - partialReviewer 3Compound 1 up-regulates HSP70 and down-regulates Bax, contributing to an anti-apoptotic gastroprotective mechanism.Qualitative immunohistochemistry images show differential staining, but no quantitation or statistics are provided for the IHC, so the claim is only partially backed.Evidence: Fig 6 and Fig 7 photomicrographs (qualitative).
HSP-70 expression was up regulated in compound 1 pretreated-animals more than that observed in ulcer control group.
Resultsreviewer’s wording - supportedReviewer 1Compound 1 possessed an excellent antioxidant activity among the compounds studied.The DPPH and FRAP data directly show compound 1 has the highest activity (IC50 0.69 µg/ml, FRAP 8966.7 µM/100g), significantly outperforming controls.Evidence: Table 1: DPPH IC50 = 0.69 ± 0.08 µg/ml, FRAP = 8966.7 ± 0.03 µM/100g, compared to other compounds and positive controls.
“Compounds 1 and 3 showed the highest radical scavenging activities, with minimum inhibitory concentration IC 50 values of (0.69 ± 0.08 μg/ml), and (2.52 ± 0.11 μg/ml), respectively, which are higher than the values obtained for the positive controls.”
Results - supportedReviewer 1Compound 1 did not cause any sign of acute toxicity in rats.The acute toxicity study at 500 mg/kg showed no deaths, no behavioral changes, and no significant differences in serum biochemistry or histology.Evidence: Acute toxicity study: 'All the animals remained alive and did not manifest any significant visible signs of toxicity at these doses.' Tables 2 and 3 show no significant differences between groups.
All the animals remained alive and did not manifest any significant visible signs of toxicity at these doses. Thus, clinical observations, serum biochemistry, and histopathology data did not show any significant differences between control and treated groups.
Resultsreviewer’s wording - supportedReviewer 1Compound 1 could significantly preserve the gastric mucosa against ethanol-induced injury.The ulcer area data show significant reduction in both compound 1 groups compared to ulcer control, with inhibition percentages of 92.6% and 97.8%.Evidence: Table 4: Ulcer area for compound 1 (50 mg/kg) = 63.36 ± 34.80 mm, (100 mg/kg) = 18.72 ± 5.42 mm, vs control = 850.00 ± 14.43 mm. Histology also shows protection.
Results showed that rats pre-treated with omeprazole, or compound 1 before being given absolute alcohol, had significantly reduced areas of gastric ulcer formation compared to rats pre-treated with (ulcer control group).
Resultsreviewer’s wording - supportedReviewer 2A series of new 2-(ethylthio)benzohydrazone derivatives (1–6) were prepared and characterised.The paper provides synthesis details and spectroscopic characterization (IR, NMR, mass) for all six compounds.Evidence: Synthesis and characterization data are provided in the Experimental section and Supplementary Information.
“The condensation reaction of 2-(ethylsulfanyl)benzohydrazide with substituted aromatic aldehydes in the presence of ethanol afforded the novel 2-(ethylthio)benzohydrazone derivatives (1–6) (). The structure of the hydrazones was confirmed by IR, 1H NMR, 13C NMR, and mass spectroscopy.”
Results and discussion - supportedReviewer 2Compound 1 has the most powerful antioxidant activity among the synthesized compounds.DPPH and FRAP assays show compound 1 has the lowest IC50 and highest FRAP value, outperforming positive controls.Evidence: Table 1 shows compound 1 has IC50 0.69 ± 0.08 μg/ml and FRAP 8966.7 ± 0.03 μM/100g.
“Compounds 1 and 3 showed the highest radical scavenging activities, with minimum inhibitory concentration IC 50 values of (0.69 ± 0.08 μg/ml), and (2.52 ± 0.11 μg/ml), respectively, which are higher than the values obtained for the positive controls.”
Results and discussion - supportedReviewer 2Compound 1 pre-treatment increases gastric pH and mucus production in ethanol-induced gastric ulcer rats.Table 4 shows significantly higher pH and mucus weight in compound 1-treated groups compared to ulcer control.Evidence: Table 4: pH of gastric tissue for compound 1 (50 mg/kg) = 5.02 ± 0.48, mucus weight = 0.50 ± 0.098 g; ulcer control pH = 2.95 ± 0.40, mucus = 0.512 ± 0.015 g.
The acidity of the gastric content in experimental rats pretreated with compound 1 was decreased significantly compared to that of the ulcer control group (p<0.05). The mucus production of gastric mucosa was significantly (p<0.05) increased in animals pretreated with compound 1 compared to the ulcer control group.
Resultsreviewer’s wording - supportedReviewer 2Compound 1 pre-treatment up-regulates HSP70 and down-regulates Bax protein expression in gastric tissue.Immunohistochemistry images and text describe increased HSP70 staining and decreased Bax staining in compound 1-treated groups.Evidence: Figures 6 and 7 show immunohistochemical staining; text states up-regulation of HSP70 and down-regulation of Bax.
HSP-70 expression was up regulated in compound 1 pretreated-animals more than that observed in ulcer control group (). Meanwhile, there was down regulation of Bax protein expression in compound 1 pretreated-animals compared to the ulcer control group ().
Resultsreviewer’s wording - supportedReviewer 2The gastroprotective effect of compound 1 may be due to its antioxidant activity and/or modulation of HSP70 and Bax.The paper presents correlative evidence linking antioxidant activity, HSP70 up-regulation, and Bax down-regulation to gastroprotection, though causality is not proven.Evidence: The paper shows compound 1 has antioxidant activity in vitro and modulates HSP70 and Bax in vivo, and discusses these as potential mechanisms.
“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 - supportedReviewer 3Compound 1 protects the gastric mucosa against ethanol-induced injury in rats.The presented ulcer-area data (reduced ulcer area and high inhibition % at both doses vs. ulcer control) and gross/histological findings directly support gastroprotection.Evidence: Table 4: ulcer area 850.00±14.43 (control) vs 63.36±34.80 (50 mg/kg) and 18.72±5.42 (100 mg/kg); Fig 3 gross photomicrographs.
“Rats pre-treated with compound 1 showed increased in gastric pH and gastric mucus.”
Abstract
Data authenticity concerns
1 finding · worst lowAn 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).
- lowdata too cleanThe inhibition percentages for compound 1 (92.6% and 97.8%) are very high, even surpassing the reference drug omeprazole (79%), which might be considered implausibly effective for a single dose without statistical comparison to omeprazole.
Compound 1 (50 mg/kg) ... 92.6 ... Compound 1 (100 mg/kg) ... 97.8 ... Omeprazole (20mg/kg) ... 79
Table 4reviewer’s wording - lowdata too cleanThe acute toxicity liver function table reports a 'TB' value of '2.00 ± 0.006' where the SD (0.006) is implausibly small relative to the mean on a μmol/L scale, and the vehicle is '2.00 ± 0.00' — suspiciously identical means across groups.
TB (μmol/L) | 2.00 ± 0.00 | ... | 2.00 ± 0.006
Table 3reviewer’s wording
Reporting gaps
4 findings · worst highRequired 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
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.
“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.”
“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 [, ].”
“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 [].”
“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.”
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.
“The rats were divided randomly into 5 groups of 6 rats each.”
“The rats were divided randomly into 5 groups of 6 rats each.”
“The rats were divided randomly into 5 groups of 6 rats each.”
“The reference group received oral doses of 20 mg/kg omeprazole in 10%Tween 20 (5 ml/kg) as positive control.”
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.
“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.”
“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.”
“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.”
“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.”
“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 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.
“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).”
“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).”
“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).”
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.
“Both proteins were purchased from Santa Cruz Biotechnology, Inc., California, USA.”
“In this study, omeprazole was used as the reference anti-ulcer drug, and was obtained from the University Malaya Medical Centre Pharmacy.”
“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.”
“Both proteins were purchased from Santa Cruz Biotechnology, Inc., California, USA.”
“All chemicals and solvents were of analytical grade and purchased from Aldrich and Merck.”
“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.”
“Both proteins were purchased from Santa Cruz Biotechnology, Inc., California, USA.”
“assessed using a one-way analysis of variance (ANOVA) followed by post hoc analysis by the prism statistical package.”
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.
“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.”
“A value of p<0.05 was considered significant.”
“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.”
“Values are assumed as mean ± S.E.M. The statistical analysis was assessed with one-way ANOVA (post hoc analysis) with P < 0.05.”
“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.”
“The statistical analysis was assessed with one-way ANOVA (post hoc analysis) with P < 0.05.”
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.
“All relevant data are within the paper and its Supporting Information files.”
“Data Availability All relevant data are within the paper and its Supporting Information files.”
“Data Availability All relevant data are within the paper and its Supporting Information files.”
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.
“Competing Interests: The authors have declared that no competing interests exist.”
“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.”
“Competing Interests: The authors have declared that no competing interests exist.”
“Competing Interests: The authors have declared that no competing interests exist.”
Broken references and links
1 finding · worst mediumReferences 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.
- datahttp://umresearch.um.edu.my/mainpage.php?module=Maklumat&kategori=86&id=63&papar=1DEADHTTP 404Dead link — nothing to verify.
Copyediting
15 minorWording, 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.
- MINORconsistencyTable 1 footnote vs. Statistical analysis“In Table 1, footnote says 'Each value represents mean ± SD', but the Statistical analysis section says 'All values were reported as mean ± S.E.M.'”→ Reconcile the reporting of dispersion; either use SD or SEM consistently, or clarify which applies to which data.This inconsistency could confuse readers.
- MINORtypoAbstract, line 1“Nazarbahjat Nafal”→ Check if this is a name or a typo; likely should be 'Nafal Nazarbahjat' or similar.The author list appears to have a formatting issue.
- MINORclarityResults, pH of gastric content and mucus production“The acidity of the gastric content in experimental rats pretreated with compound 1 was decreased significantly compared to that of the ulcer control group (p<0.05).”→ Specify which dose of compound 1 (50 or 100 mg/kg) is being compared, or present both.The sentence is ambiguous.
- MINORpunctuationThroughout the text“Missing spaces after periods in some places (e.g., '...in rats.Adult healthy...')”→ Add spaces after periods throughout the manuscript.Minor formatting issues.
- MINORgrammarAbstract“increased in gastric pH”→ increased gastric pHPreposition 'in' is unnecessary.
- MINORtypoDiscussion“flatting of the gastric mucosal folds”→ flattening of the gastric mucosal foldsCorrect spelling is 'flattening'.
- MINORtypoResults, Mucus (PAS) staining“Upsurge in the glycogen content”→ Increase in the glycogen content'Upsurge' is less common; 'increase' is clearer.
- MINORpunctuationResults, Gross pathological findings“the standard drug used to treat gastric ulcer, and”→ the standard drug used to treat gastric ulcer.Incomplete sentence trailing 'and'.
- MINORclarityResults, DPPH Radical scavenging activity“Compounds 1 and 3 showed the highest radical scavenging activities, with minimum inhibitory concentration IC 50 values of (0.69 ± 0.08 μg/ml), and (2.52 ± 0.11 μg/ml), respectively, which are higher than the values obtained for the positive controls.”→ Compounds 1 and 3 showed the highest radical scavenging activities, with IC50 values of 0.69 ± 0.08 μg/ml and 2.52 ± 0.11 μg/ml, respectively, which are lower than those of the positive controls.Higher scavenging activity corresponds to lower IC50; clarify comparison.
- MINORtypoMaterials and Methods, Gastric ulcer study/Table 4“10% Twee20”→ 10% Tween 20Missing 'n' in Tween; recurring typo.
- MINORtypoTable 4 header“pH of Gastrictissue”→ pH of gastric tissueMissing space.
- MINORclarityResults, DPPH section“with minimum inhibitory concentration IC 50 values”→ with IC50 valuesIC50 is not a 'minimum inhibitory concentration' in this antioxidant context; avoid the MIC terminology.
- MINORgrammarResults, Discussion“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”→ ...and even resistance to these drugs is emergingSubject-verb agreement ('resistance ... is').
- MINORtypoMaterials and Methods, FRAP assay“1 ml of the2,4,6-tripyridyl-s-triazine)TPTZ solution”→ 1 ml of the 2,4,6-tripyridyl-s-triazine (TPTZ) solutionMissing space and stray parenthesis.
- MINORclarityResults, Table 1 text“rutin (394)”→ rutin (393.89)Inconsistent rounding vs. table value 393.89±0.02.
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.HIGHrigorAcknowledge 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.HIGHstatisticsProvide 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 codeDisclose 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.HIGHotherClarify 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.HIGHrigorState 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.HIGHreportingAdd 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.HIGHreportingReference the ARRIVE guidelines for animal research in the Methods or Ethics statement.Adherence to reporting guidelines is expected for preclinical animal studies.
- 8.HIGHdata codeDeposit 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.MEDIUMcopyeditReconcile 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.MEDIUMcopyeditCorrect the typo '10% Twee20' to '10% Tween 20' throughout the manuscript.Typographical errors can undermine confidence in the reporting.
- 11.MEDIUMcopyeditIn 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.LOWcopyeditCorrect 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.
The star rating is the report’s one-glance summary. Every paper starts at 5★ and loses stars for the concrete problems the review finds — so a rating is never a vague average, it’s a running total you can read line by line under “How this rating was calculated.”
- Reporting — 8 dimensionseach dimension that fully fails−½★
- each dimension partially met−¼★
- Statistics · Integrity · Claimseach serious problem−1★
- each medium problem−½★
- Citationseach retracted or unverifiable reference−¼★
- Copyeditonly when the manuscript needs a full edit−½★
The rating never drops below 1★, and a demonstrable critical failure (an impossible statistic, a proven ethics violation) caps it at 1★ on its own — so the stars can never look healthy when the verdict is CRITICAL.
The rating draws on a panel of agents. Three independent Kaimen Rigor reviewers grade the eight dimensions below across several independent passes (the shown verdict is their majority vote — steadier than any single run), isolate the paper’s major claims and check its own evidence backs them, and flag integrity concerns. Alongside them, a citation agent resolves every reference against Crossref, OpenAlex, and Retraction Watch; a statistics agent recomputes reported tests; and rule-based checks verify that declared data/code links actually resolve. Full text is required — an abstract-only submission is not analyzed.
Graded against NIH, MDAR, ARRIVE 2.0, CONSORT, EQUATOR, and RRID guidelines. A dimension that doesn’t apply to the study type is skipped, never penalized.
This Kaimen Rigor review is model-assisted and is not a substitute for formal expert review. It complements human evaluation by surfacing potential methodological concerns — verify each finding against the source.