Rigor Reviewed

Rigor-Reviewed Manuscript

Chronic administration of R-flurbiprofen attenuates learning impairments in transgenic amyloid precursor protein mice

Name: Adcurare Rigor Record — Chronic administration of R-flurbiprofen attenuates learning impairments in transgenic amyloid precursor protein mice
Item: Chronic administration of R-flurbiprofen attenuates learning impairments in transgenic amyloid precursor protein mice
Rating: 39
Author: Adcurare

Kukar T, Prescott S, Eriksen JL, ... Koo EH, Golde TE

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Adcurare Rigor Review — 1★ (Critical), 2 critical, 6 high issues for “Chronic administration of R-flurbiprofen attenuates learning impairments in transgenic amyloid precursor protein mice”

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Adcurare Editorial

This rigor review was examined and confirmed by Adcurare Editorial · July 28, 2026

Adcurare Rigor Review

Rigor Reviewed

1 / 5

Critical — a demonstrable failure

Interventional Study

Critical

High-priority

How this rating was calculated▸

Started at 5★ — deductions

IntegrityIntegrity concern ×5−2.5★
StatisticsImpossible or misreported statistic ×2−2★
StatisticsStatistic did not reproduce ×3−1.5★
ReportingKey resources not met−0.5★
ReportingStatistical analysis not met−0.5★
ReportingData & code availability not met−0.5★
ReportingStudy design partially met−0.25★

A demonstrable critical failure caps the rating at the minimum, regardless of the deductions above.

Model-reported confidence: highReviewer agreement: highEngine: 7.7.0Source: full text

Not verified in this report

References were not verified against Crossref/OpenAlex.
No data or code availability links were detected to verify.

Problems surfaced8 types

Significance claim flips when recomputed
Statistical analysis inadequately reported
Data and code not shared
Key resources not identified
Reported statistics do not recompute
Study-design details incomplete (controls, blinding, power)
Internal contradictions in the reported numbers
Other integrity concern

This Adcurare 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.

Critical findings

• Recomputed 13 tests: 8 consistent, 5 inconsistent (2 change significance at p<.05), 13 via agent-written checks.

The paper has a strong scientific premise, adequate ethical approvals, and generally transparent reporting, but it is undermined by major gaps in study design (randomization, blinding), incomplete identification of key resources, deficient statistical reporting (including verified inconsistencies and a rounding error), and a complete absence of data/code availability. These issues collectively reduce confidence in the published findings and warrant a correction or re-analysis.

Three independent reviewers (all AI) fully agreed on dimension statuses, strengthening reliability. The statistics verification component recomputed 13 tests (coverage limited to those with test statistic+df or effect+CI), finding 5 inconsistencies and 2 decision errors. The copyedit pass flagged 11 minor issues. No citation concerns were raised. The study is a published preclinical interventional paper; this assessment is a post-publication audit.

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

Recomputed 13 tests: 8 consistent, 5 inconsistent (2 change significance at p<.05), 13 via agent-written checks.

5 integrity concerns flagged (0 high).

Reporting

Eight Dimensions4 pass · 1 warn · 3 fail

The background cites epidemiological and experimental evidence linking NSAIDs to AD risk and Aβ42 lowering, including prior work on R-flurbiprofen. The hypothesis that R-flurbiprofen could improve cognition and reduce Aβ pathology is logically derived. Limitations of prior clinical trials and the need for further mechanistic studies are acknowledged.

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 premise by citing relevant prior work, providing a logical rationale, and addressing limitations of previous studies.

Evidence

direct quote[Background, paragraph 1]

“We and others have shown that certain NSAIDs reduce secretion of Aβ42 in cell culture and animal models, and that the effect of NSAIDs on Aβ42 is independent of the inhibition of cyclooxygenase by these compounds.”

direct quote[Background, paragraph 3]

“The identification of compounds that selectively lower Aβ42 provides a unique opportunity to test the hypothesis that decreasing levels of this peptide will have a positive impact on Aβ plaque pathology and cognition.”

direct quote[Background, paragraph 4]

“Unfortunately, the potential toxicity related to inhibition of COX as well as diverse secondary targets of this class of drugs, complicates efforts to experimentally test this hypothesis.”

PASSReviewer 2· 95% conf

The scientific premise is well-established with extensive prior work cited, a clear logical rationale, and acknowledgment of limitations of prior research.

Evidence

direct quote[Background, paragraph 3-4]

“Over the past 20 years, a substantial number of epidemiological reports have shown that long-term use of non-aspirin, nonsteroidal, anti-inflammatory drugs (NSAIDs) is associated with protection from the development of AD”

direct quote[Background, paragraph 6]

“Extensive screening of NSAIDs, derivatives and related compounds led to the identification of R -flurbiprofen as a promising selective Aβ42-lowering agent that may circumvent some of these complications.”

direct quote[Background, paragraph 5]

“Unfortunately, the potential toxicity related to inhibition of COX as well as diverse secondary targets of this class of drugs, complicates efforts to experimentally test this hypothesis.”

PASSReviewer 3· 95% conf

The paper establishes a strong scientific premise by citing relevant prior work on NSAIDs in AD, the hypothesis that Aβ42 lowering may be protective, and the rationale for testing R-flurbiprofen.

Evidence

paraphrase[Background, paragraphs 1-5]

“The paper reviews epidemiological studies, animal model work, and the discovery that certain NSAIDs lower Aβ42 independently of COX inhibition, leading to the rationale for testing R-flurbiprofen.”

direct quote[Background, paragraph 5]

“Based on R -flurbiprofen's selective lowering of Aβ42, reduced COX activity and safety profile in humans we have previously suggested that this drug was a good candidate for clinical testing in AD.”

direct quote[Background, paragraph 3]

“Given the growing AD epidemic there is an urgent need to discover alternative, more effective, therapies that not only target the disease symptoms but can also slow or halt the underlying neurodegenerative process.”

No randomization method is described; mice were assigned to groups based on age. Blinding was only reported for plaque quantification, not for drug administration or behavioral testing. No a priori power analysis was performed. Inclusion/exclusion criteria were partially defined (cue training performance, stereotypic behavior). Outlier handling for behavioral data was reported but not for biochemical assays. Controls (untreated mice) were included, and the study was replicated across two experiments (1 and 2).

Sub-criteria

controlsADEQUATE

power analysisNOT REPORTED

blinding levelsINADEQUATE

outlier handlingINADEQUATE

randomization unitNOT REPORTED

inclusion exclusionADEQUATE

randomization methodNOT REPORTED

replicate distinctionNOT REPORTED

independent replicationADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 85% conf

The study design has several reporting gaps: randomization method, blinding, and power analysis are not reported; replication and inclusion/exclusion criteria are adequate.

Evidence

direct quote[Methods, R-Flurbiprofen experiments]

“The mice were assigned to a 'Preventative' or a 'Therapeutic' experimental group based on age.”

direct quote[Methods, Spatial learning assessment]

“Mice that performed +/- three times the standard deviation from the mean were excluded from further study.”

direct quote[Methods, Statistical analysis]

“Experiments 1 and 2 were replicate experiments investigating the long-term treatment effects of R-flurbiprofen on spatial memory.”

WARNReviewer 2· 85% conf

The study design lacks randomization, blinding (except for plaque quantification), and a power analysis, though inclusion/exclusion criteria and outlier handling are well-defined.

Evidence

direct quote[Methods, 'R-Flurbiprofen experiments']

“The mice were assigned to a 'Preventative' or a 'Therapeutic' experimental group based on age.”

direct quote[Methods, 'Quantitation of amyloid plaque burden']

“Computer-assisted quantification of Aβ plaques was performed in a blinded fashion”

direct quote[Methods, 'Spatial learning assessment']

“Mice that performed +/- three times the standard deviation from the mean were excluded from further study.”

WARNReviewer 3· 90% conf

The study design has several strengths, including pre-specified exclusion criteria and attempted replication, but lacks randomization, blinding at multiple stages, power analysis, and clear distinction between biological and technical replicates.

Evidence

absence[Methods, subsection 'R-Flurbiprofen experiments']

No randomization method described.

direct quote[Methods, subsection 'Quantitation of amyloid plaque burden']

“Computer-assisted quantification of Aβ plaques was performed in a blinded fashion”

direct quote[Methods, subsection 'Spatial learning assessment']

“Mice that performed +/- three times the standard deviation from the mean were excluded from further study.”

The use of female Tg2576 mice is stated, but no scientific justification for single-sex is provided. Age at treatment onset and sacrifice are reported; weight and health status are not explicitly described. The strain (C57BL/6.SJL background) and breeding scheme are detailed. Housing (3-5 per cage, food/water ad lib, 12h light/dark) is described.

Sub-criteria

demographicsN/A

sex reportedADEQUATE

sex justifiedNOT REPORTED

age weight healthINADEQUATE

housing conditionsADEQUATE

species strain sourceADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 90% conf

Sex, species/strain, and housing are adequately reported; age is given but weight/health not detailed; no justification for single-sex use.

Evidence

direct quote[Methods, Animals]

“Female Tg2576 mice were generated and confirmed by genotyping as described previously.”

direct quote[Methods, Animals]

“All animals were housed 3-5 to a cage and maintained on ad libitum food and water with a 12-hour light/dark cycle in a controlled environment.”

PASSReviewer 2· 90% conf

Sex, age, species/strain, and housing conditions are adequately reported, but no justification is given for using only female mice.

Evidence

direct quote[Methods, 'Animals']

“Female Tg2576 mice were generated and confirmed by genotyping as described previously”

direct quote[Methods, 'Animals']

“Kibble consumption, general health and body weight were monitored on a weekly basis.”

direct quote[Methods, 'Animals']

“All animals were housed 3–5 to a cage and maintained on ad libitum food and water with a 12-hour light/dark cycle in a controlled environment.”

PASSReviewer 3· 85% conf

Sex, strain, and housing conditions are adequately reported, but justification for using only females is missing, and age/health reporting is somewhat vague.

Evidence

direct quote[Methods, subsection 'Animals']

“Female Tg2576 mice were generated and confirmed by genotyping”

direct quote[Methods, subsection 'R-Flurbiprofen experiments']

“Kibble consumption, general health and body weight were monitored on a weekly basis.”

direct quote[Methods, subsection 'Animals']

“All animals were housed 3–5 to a cage and maintained on ad libitum food and water with a 12-hour light/dark cycle in a controlled environment.”

The methods state that all procedures were approved by the Mayo Clinic Institutional Animal Care and Use Committee and conducted in accordance with NIH guidelines, meeting the criteria for ethical reporting in animal research.

Sub-criteria

iacuc statementADEQUATE

informed consentN/A

irb ethics statementN/A

regulatory complianceADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 95% conf

IACUC approval and regulatory compliance are adequately stated.

Evidence

direct quote[Methods, Animals]

“All animal husbandry and testing procedures performed were approved by the Mayo Clinic Institutional Animal Care and Use Committee in accordance with NIH guidelines.”

PASSReviewer 2· 95% conf

Animal ethics approval is clearly stated with a named committee and compliance with NIH guidelines.

Evidence

direct quote[Methods, 'Animals']

“All animal husbandry and testing procedures performed were approved by the Mayo Clinic Institutional Animal Care and Use Committee in accordance with NIH guidelines.”

PASSReviewer 3· 95% conf

Ethical approvals and regulatory compliance for animal research are adequately reported.

Evidence

direct quote[Methods, subsection 'Animals']

“All animal husbandry and testing procedures performed were approved by the Mayo Clinic Institutional Animal Care and Use Committee in accordance with NIH guidelines.”

Antibodies used (e.g., 33.1.1, anti-Iba1, anti-GFAP) are named but not with complete vendor/catalog/RRID information. R-flurbiprofen source is not provided; only the diet formulation company is named. The Tg2576 mouse strain is described but no RRID or commercial source given. Software (HVS Image, MetaMorph) is adequately identified with versions.

Sub-criteria

mycoplasma testingN/A

reagents identifiedINADEQUATE

organisms identifiedINADEQUATE

antibodies identifiedINADEQUATE

cell line authenticationN/A

software tools identifiedADEQUATE

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 85% conf

Key resources for antibodies and the investigational drug are inadequately identified; software tools lack versions and RRIDs; no catalog numbers or authentication provided.

Evidence

direct quote[Methods, Analysis of Aβ levels]

“To measure Aβ42 levels-peptides were captured with mAb 2.1.3 (specific for the c-terminus of Aβ42) and detected with HRP-conjugated mAb Ab9 (specific for human Aβ1-16).”

direct quote[Methods, R-Flurbiprofen experiments]

“R-Flurbiprofen, at a concentration of 67, 167.5 or 335 or mg per kg of diet was homogenously incorporated into Harlan Teklad 7012 kibble by Research Diets, Inc.”

direct quote[Methods, Spatial learning assessment]

“Data are analyzed using a video tracking system and software developed by Richard Baker, HVS Image Analyzing (Hampton, U.K.).”

FAILReviewer 2· 85% conf

Key resources are inadequately identified: antibodies lack vendor/catalog numbers, software lacks version details, and the drug is not fully specified with a catalog number.

Evidence

direct quote[Methods, 'Immunohistology' and 'Analysis of Aβ levels']

“To measure Aβ42 levels-peptides were captured with mAb 2.1.3 (specific for the c-terminus of Aβ42) and detected with HRP-conjugated mAb Ab9”

direct quote[Methods, 'R-Flurbiprofen experiments']

“R -Flurbiprofen, at a concentration of 67, 167.5 or 335 or mg per kg of diet was homogenously incorporated into Harlan Teklad 7012 kibble by Research Diets, Inc.”

direct quote[Methods, 'Spatial learning assessment']

“Data are analyzed using a video tracking system and software developed by Richard Baker, HVS Image Analyzing (Hampton, U.K.).”

FAILReviewer 3· 90% conf

Key biological resources are inadequately reported: antibodies lack full identifiers, drug source is not specified, and mouse strain lacks RRID.

Evidence

paraphrase[Methods, subsections 'Analysis of Aβ levels' and 'Immunohistology']

“Antibodies are named (e.g., 2.1.3, Ab9, 13.1.1, 33.1.1, anti-Iba1, anti-GFAP) but vendors and catalog numbers are only given for Iba1 and GFAP; no RRIDs.”

direct quote[Methods, subsection 'R-Flurbiprofen experiments']

“R -Flurbiprofen, at a concentration of 67, 167.5 or 335 or mg per kg of diet was homogenously incorporated into Harlan Teklad 7012 kibble by Research Diets, Inc.”

direct quote[Methods, subsection 'Quantitation of amyloid plaque burden']

“MetaMorph 6.1 software (Universal Imaging Corp, Downington, PA)”

Statistical tests (repeated measures ANOVA, Fisher's PLSD, Chi-square) are named. However, no verification of ANOVA assumptions (normality, homogeneity of variance) is reported. Many p-values are reported only as thresholds (e.g., p < 0.05, p < 0.0001) rather than exact values. No effect sizes or confidence intervals are provided. The statistical software used is not identified. Data are presented as bar graphs without individual data points, and error bars (SEM) are not always defined in figure legends.

Sub-criteria

tests namedADEQUATE

exact p valuesINADEQUATE

effect sizes ciNOT REPORTED

data presentationINADEQUATE

software identifiedNOT REPORTED

assumptions verifiedNOT REPORTED

mathematical plausibilityPASS

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 80% conf

Statistical reporting is deficient: test assumptions not verified, exact p-values not consistently reported, effect sizes and CIs absent, statistical software not identified, and data presentation lacks individual points.

Evidence

direct quote[Results, Spatial reference memory]

“A significant interaction between training day and group indicated that the groups differed on certain days but there was no main effect of drug-treatment group [F(2,55) = 1.16, ns].”

absence[Results (all sections)]

Effect sizes and CIs not reported.

absence[Methods, Statistical analysis]

Statistical software not identified.

FAILReviewer 2· 80% conf

Statistical reporting is incomplete: test assumptions are not verified, exact p-values are rarely given, effect sizes/confidence intervals are absent, and the statistical software is not identified.

Evidence

absence[Methods, 'Statistical analysis']

No verification of ANOVA assumptions (normality, equal variance) is reported.

direct quote[Results, e.g., 'spatial bias' paragraph]

“main effect of group [F(2,55) = 3.91, p < .05]”

direct quote[Results, 'Strategy preference']

“31% of the Therapeutic group”

FAILReviewer 3· 85% conf

Statistical reporting is inadequate: tests are named but assumptions not verified, exact p-values are often missing, effect sizes not reported, software not identified, and data presentation lacks individual points.

Evidence

direct quote[Methods, subsection 'Statistical analysis']

“Repeated measures ANOVAs (× Group) were then used to examine visible platform latency, hidden platform search error and probe trial data.”

direct quote[Results, multiple locations]

“p < .0001 (e.g., for plaque burden reduction)”

absence[Results, throughout]

No effect sizes or confidence intervals reported.

The paper does not include a data availability statement. Raw data are not deposited in a public repository. No accession numbers for sequencing or other data are provided. No custom code was shared or mentioned.

Sub-criteria

code sharingNOT REPORTED

accession numbersNOT REPORTED

repository depositNOT REPORTED

data availability statementNOT REPORTED

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 95% conf

No data availability statement, no repository deposit, no accession numbers, and no shared code.

Evidence

absence[Entire paper]

No data availability statement.

FAILReviewer 2· 90% conf

No data availability statement is provided, and no data or code repositories are mentioned.

Evidence

absence[Entire manuscript]

No data availability statement is present anywhere in the manuscript.

FAILReviewer 3· 95% conf

No data or code availability statements are provided; no public repositories are mentioned.

Evidence

absence[Entire paper]

No data availability statement.

The methods provide sufficient detail for replication of drug administration, behavioral testing, and biochemical analyses. All prespecified outcomes (cognition and Aβ) are reported. Limitations such as sub-optimal dosing, conversion to S-enantiomer, and variability in cognitive phenotype are discussed. Conclusions are cautious and proportional. However, no reporting guideline (e.g., ARRIVE) is referenced, and a conflict of interest statement is absent.

Sub-criteria

funding coiNOT REPORTED

trial registrationN/A

reporting guidelineNOT REPORTED

methods completenessADEQUATE

all outcomes reportedADEQUATE

limitations discussedADEQUATE

conclusions proportionalADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 90% conf

Methods are detailed, all outcomes are reported, limitations are discussed, and conclusions are proportional; but no reporting guideline checklist and no COI statement.

Evidence

direct quote[Methods (entire)]

“R-Flurbiprofen, at a concentration of 67, 167.5 or 335 or mg per kg of diet was homogenously incorporated into Harlan Teklad 7012 kibble by Research Diets, Inc.”

direct quote[Discussion and conclusion, paragraph 5]

“An important caveat is that these results were obtained using a sub-optimal dose of R-flurbiprofen, which was necessitated by the toxicity of the compound at higher doses in these mice.”

absence[End of paper]

No COI statement.

PASSReviewer 2· 85% conf

Methods are detailed, all outcomes are reported, limitations are discussed, and conclusions are proportional, but no reporting guideline is referenced and a formal conflicts of interest statement is missing.

Evidence

direct quote[Acknowledgements]

“These studies were funded by the US National Institutes of Health National Institute on Aging and the Alzheimer's Association”

direct quote[Discussion, paragraph 4]

“An important caveat is that these results were obtained using a sub-optimal dose of R -flurbiprofen, which was necessitated by the toxicity of the compound at higher doses in these mice”

absence[Methods and entire manuscript]

No reporting guideline is referenced.

PASSReviewer 3· 90% conf

Reporting is generally transparent: methods are detailed, outcomes are fully reported, limitations are discussed, conclusions are proportional, and funding is acknowledged; however, a formal conflicts of interest statement is missing.

Evidence

direct quote[Discussion, final paragraph]

“An important caveat is that these results were obtained using a sub-optimal dose of R -flurbiprofen, which was necessitated by the toxicity of the compound at higher doses in these mice, most likely caused by bioinversion to S -flurbiprofen”

direct quote[Acknowledgements]

“These studies were funded by the US National Institutes of Health National Institute on Aging and the Alzheimer's Association (P01 AG20206 to E.K. and T.G.; NIRG 02-4082 to M.M.N.).”

11 copyedit issues flagged: mostly typo, consistency, clarity.

Adcurare Rigor Action ItemsFinal pass before journal submission

As a published paper, the robustness is compromised by the statistical inconsistencies (5 of 13 tests inconsistent, including 2 decision errors) and a mathematical plausibility issue (31% of 15 mice is non-integer). An informed reader should weigh whether these errors affect the paper's conclusions. The authors should consider issuing a correction or providing a re-analysis with corrected p-values and clarification of the rounding error. The missing data availability statement limits reproducibility.

1.
CRITICALstatistics
Re-analyze the 5 inconsistent tests (including the 2 decision errors) identified by the statistics verification component, report corrected p-values, and re-evaluate whether conclusions change.
5 of 13 recomputed tests were inconsistent with reported values, and 2 p-values reported as significant were non-significant on recomputation, indicating a potential data analysis error that could affect the paper's conclusions.
2.
CRITICALstatistics
Clarify the rounding error in the strategy preference result: 31% of 15 mice is 4.65, not an integer. Provide exact counts or state that the percentage is an approximation.
This inconsistency suggests a potential data reporting error or miscalculation that undermines the credibility of the reported result.
3.
HIGHdata code
Add a data availability statement specifying how raw behavioral and biochemical data can be accessed (e.g., on request from the authors or via a repository).
The current paper provides no way for readers to access the underlying data, which is a major barrier to reproducibility and verification.
4.
HIGHrigor
Provide full identifiers (vendor, catalog number, RRID) for all antibodies and key reagents, including the source of R-flurbiprofen (manufacturer, catalog number, purity).
Without these identifiers, other researchers cannot reproduce the experiments or verify the reagents used.
5.
HIGHrigor
Describe the randomization method used to assign mice to treatment groups, or state clearly why randomization was not feasible.
The current assignment based on age introduces potential confounding and is a significant methodological limitation.
6.
HIGHrigor
Report whether investigators were blinded to treatment group during behavioral testing and data analysis, or state clearly why blinding was not performed.
Blinding is a standard safeguard against bias; its absence is a major concern for the validity of behavioral results.
7.
HIGHstatistics
Report exact p-values (e.g., p = 0.023) for all statistical tests instead of threshold values, and include effect sizes (e.g., Cohen's d) and 95% confidence intervals for primary outcomes.
Threshold p-values are insufficient for precise interpretation, and effect sizes are essential for understanding the magnitude of effects.
8.
HIGHstatistics
Identify the statistical software used (e.g., SPSS, GraphPad Prism) with version number in the Methods section.
The software used can affect the reproducibility of analyses, and its absence is a reporting gap.
9.
MEDIUMreporting
Add a formal conflicts of interest statement, even if stating that the authors declare no competing interests.
Transparency about potential conflicts is a standard requirement for most journals.
10.
MEDIUMreporting
Reference the ARRIVE guidelines for reporting animal research, and provide a completed checklist as supplementary material.
Adherence to reporting guidelines is recommended by many journals and improves the completeness and transparency of the report.
11.
MEDIUMcopyedit
Correct the typo in Table 2 header: 'Tg2575' should be 'Tg2576'.
This is a clear typographical error that could cause confusion about the mouse strain used.
12.
MEDIUMcopyedit
Standardize the drug name to 'R-flurbiprofen' throughout the manuscript (remove extra spaces and hyphens).
Inconsistent formatting of the drug name is distracting and may appear unprofessional.
13.
MEDIUMrigor
Provide a scientific justification for using only female mice, or discuss the implications of this limitation.
Single-sex studies without justification may limit the generalizability of the findings.
14.
LOWreporting
Consider showing individual data points in all figures (e.g., dot plots) alongside bar graphs to reveal distribution and variability.
Bar graphs with error bars can obscure the underlying data distribution; individual points improve transparency.

Full text (61,093 chars)Analyzed: Jul 28, 2026

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