Protein expression analysis by western blots appears to be the weak spot of life science, because these blots are so often reported to be manipulated. However, this is only because anybody, even without any background in biology can spot re-used western blot bands, just using a good eye or some computer skills. Unlike with other analytical tools, like real-time PCR or microscopy imaging, you don’t need specialist knowledge or access to raw data to spot digital manipulations of western blots. One of the most famous western blot breeders is the Brazilian diabetes researcher Mario Saad (see whistleblower reports on my site here and here). Below is the case of his two colleagues from Poland.
Some time ago, I was contacted by a Polish scientist with a dossier of suspected western blot duplications in the publication of the pharmacologist Bożena Gabryel from the Department of Pharmacology at the Medical University of Silesia in Katowice, Poland. Gabryel specializes in oxidative stress research, four of her publications on this topic were accused to feature duplicated western blots, in different context. Sometimes suspicions were raised that selected individual bands in different western blots look too similar, which might suggest manipulative band duplications.
Soon after, another whistleblower independently contacted me with another dossier about yet another set of Gabryel’s papers, all featuring as lead or even corresponding author Krzysztof Łabuzek, who is an adjunct professor at this same Medical University of Silesia with interest in immune system and who also has a clinical practice for internal medicine in a small town near Katowice. Labuzek’s papers seem to feature a whole gallery of re-used western blots, as the dossier suggests. Additionally, that second whistleblower expressed concerns about western blot splicing and suspected band re-used in papers featuring the first author Joanna Ślusarczyk, PhD student in the lab of Agnieszka Basta-Kaim at the Polish Academy of Sciences in Krakow. Basta-Kaim is also co-author on one Gabryel paper now suspected of data manipulations.
Peculiarly, Labuzek and his coauthors had to retract in December 2016 the paper Buldak et al 2016 from Spandidos journal Experimental and Therapeutic Medicine, not even a year after that paper was published. The retraction notice shows a peculiar attitude of the authors to research integrity:
“ Following an enquiry that we received concerning the data presented in the western blots in Figs. 2-5 of the above article, we attempted to perform a thorough analysis of the data, but the original blots in their entirety were found to be missing. The error in blot selection is likely to have occurred during the final stages of figure preparation, and furthermore, the stored biological material was insufficient to enable the experiments to be repeated. Although we consider that the integrity of the data was not affected (which, unfortunately, we are unable to confirm), all the authors are in agreement that it would be prudent to retract the manuscript”.
So, to Labuzek even if the actual experimental data or samples are missing (or maybe never existed), there is no reason to doubt their hypothetical integrity anyway. One is tempted to see Labuzek alone as the creative party responsible for re-used western blots and bands, but his name is absent on two of the four Gabryel papers reported by the first whistleblower (who chose an assumed name “Ewa Stutman”). These are the papers, and the responses I obtained from the Editors-in-Chief (EiC) of the respective journals is also provided.
- Gabryel B, Jarząbek K, Machnik G, Adamczyk J, Belowski D, Obuchowicz E, Urbanek T. Superoxide dismutase 1 and glutathione peroxidase 1 are involved in the protective effect of sulodexide on vascular endothelial cells exposed to oxygen-glucose deprivation. Microvasc Res. 2016 Jan;103:26-35. doi: 10.1016/j.mvr.2015.10.001. Epub 2015 Oct 23. PubMed PMID: 26477504.
Here, the journal’s EiC Patricia A. D’Amore, professor of ophthalmology and pathology at Harvard Medical School, USA announced to “forward it to our legal department who handles such issues”.
- Urbanek T, Kuczmik W, Basta-Kaim A, Gabryel B. Rapamycin induces of protective autophagy in vascular endothelial cells exposed to oxygen-glucose deprivation. Brain Res. 2014 Mar 17;1553:1-11. doi: 10.1016/j.brainres.2014.01.017. Epub 2014 Jan 22. PubMed PMID: 24462935.
Unfortunately, the EiC of that journal, Matthew LaVoie, who is professor of neurology at Brigham and Women’s Hospital, USA, chose not to reply to my two emails. Thus, I am not sure if this Harvard Medical School professor found the Polish evidence on data integrity in his own journal compelling enough to waste his precious time on it.
- Gabryel B, Kost A, Kasprowska D, Liber S, Machnik G, Wiaderkiewicz R, Łabuzek AMP activated protein kinase is involved in induction of protective autophagy in astrocytes exposed to oxygen-glucose deprivation. Cell Biol Int. 2014 Oct;38(10):1086-97. doi: 10.1002/cbin.10299. Epub 2014 Jun 18. PubMed PMID: 24798185
The EiC of this journal, Sergio Schenkman, professor of microbiology at the Universidade Federal de Sao Paulo in Brazil, seemed rather convinced by the figure reuse across sevral manuscripts and announced to me to “ask the authors an explanation and take the right measures soon”.
- Gabryel B, Bielecka A, Bernacki J, Łabuzek K, Herman ZS. Immunosuppressant cytoprotection correlates with HMGB1 suppression in primary astrocyte cultures exposed to combined oxygen-glucose deprivation. Pharmacol Rep. 2011;63(2):392-402. PubMed PMID: 21602594.
A reaction from this journal’s EiC is presented below.
The whistleblower Ewa Stutman tells this:
“Some time ago reading original papers I noticed surprisingly similar bands in presented western blots. After looking in depth into data showed in other publications of the same corresponding author (Bozena Gabryel I found a series of publications where the same bands were repeatedly presented as results of immunodetection of different proteins in totally different experiments. I couldn’t believe my eyes when I saw that”.
Here are some of these examples, the full dossier is available here:
Ewa Stutman concluded:
“There is no problem to find more combinations of the same bands showed in different configurations in publications of Dr Gabryel. Only a few examples are presented“.
The second whistleblower offered no assumed name. Below is some of his evidence (full dossier here and here), following the list of flagged Labuzek papers, Gabryel is a co-author on four of them:
1. Łabuzek K, Liber S, Gabryel B, Okopień B. AICAR (5-aminoimidazole-4-carboxamide-1-beta-4-ribofuranoside) increases the production of toxic molecules and affects the profile of cytokines release in LPS-stimulated rat primary microglial cultures. Neurotoxicology. 2010 Jan;31(1):134-46. doi: 10.1016/j.neuro.2009.10.006.
The EiC of this journal, Joan Cranmer, professor of pediatrics at the University of Arkansas for Medical Sciences, USA, forwarded the evidence to the Elsevier Journal Manager, Clement Abinesh, who announced to me to “check the details” and keep me updated.
2. Łabuzek K, Liber S, Gabryel B, Bułdak L, Okopień B. Ambivalent effects of compound C (dorsomorphin) on inflammatory response in LPS-stimulated rat primary microglial cultures. Naunyn Schmiedebergs Arch Pharmacol. 2010 Jan;381(1):41-57. doi: 10.1007/s00210-009-0472-2.
3. Łabuzek K, Liber S, Gabryel B, Adamczyk J, Okopień B. Metformin increases phagocytosis and acidifies lysosomal/endosomal compartments in AMPK-dependent manner in rat primary microglia. Naunyn Schmiedebergs Arch Pharmacol. 2010 Feb;381(2):171-86. doi: 10.1007/s00210-009-0477-x.
Here the journal’s EiC Roland Seifert, head of Institute of Pharmacology at Hannover Medical School (MHH) in Germany promised in his email from December 11th 2016 to investigate the “suspected case of data duplication” according to their standard operating practice.
4. Łabuzek K, Liber S, Gabryel B, Okopień B. Metformin has adenosine-monophosphate activated protein kinase (AMPK)-independent effects on LPS-stimulated rat primary microglial cultures. Pharmacol Rep. 2010 Sep-Oct;62(5):827-48.
5. Łabuzek K, Liber S, Bułdak Ł, Machnik G, Liber J, Okopień B. Eplerenone promotes alternative activation in human monocyte-derived macrophages. Pharmacol Rep. 2013;65(1):226-34.
6. Bułdak Ł, Łabuzek K, Bułdak RJ, Kozłowski M, Machnik G, Liber S, Suchy D, Duława-Bułdak A, Okopień B. Metformin affects macrophages’ phenotype and improves the activity of glutathione peroxidase, superoxide dismutase, catalase and decreases malondialdehyde concentration in a partially AMPK-independent manner in LPS-stimulated human monocytes/macrophages. Pharmacol Rep. 2014 Jun;66(3):418-29. doi: 10.1016/j.pharep.2013.11.008.
7. Suchy D, Łabuzek K, Bułdak Ł, Szkudłapski D, Okopień B. Comparison of chosen activation markers of human monocytes/macrophages isolated from the peripheral blood of young and elderly volunteers. Pharmacol Rep. 2014 Oct;66(5):759-65. doi: 10.1016/j.pharep.2014.04.008.
The EiC of this Polish journal Władysław Lasoń, pharmacology professor at the Polish Academy of Sciences in Krakow, acknowledged receiving the dossiers and forwarded me this editorial decision on another Labuzek paper which was not even mentioned in the dossier:
“Because of a mistake in Fig. 3, the corresponding author (Dr. Bułdak) requested to publish a corrigendum to the paper: Łukasz Bułdak, Grzegorz Machnik, Rafał Jakub Bułdak, Krzysztof Łabuzek, Aleksandra Bołdys, Dariusz Belowski, Marcin Basiak, Bogusław Okopień “Exenatide (a GLP-1 agonist) expresses anti-inflammatory properties in cultured human monocytes/macrophages in a protein kinase A and B/Akt manner”, 2016, vol. 68(2), 329-337. The request was approved by Prof. Lasoń and Dr. Grażyna Skuza”.
These files were attached, the erratum notice which simply stated without elaborating: “in this published article, Figure 3 contained a mistake”, and the corrected figure 3.
We can probably expect at least five more corrections of the Labuzek papers flagged by the two whistleblowers from Pharmacological Reports, which will mention in passing that some figures contained “a mistake”.
I notified also the Polish university of Gabryel and Labuzek. According to my information, the internal investigations at the Medical University of Silesia were dragging on without much visible progress, part of delay was caused by Gabryel herself postponing the hearing and digging up new evidence which allegedly exonerated her. Eventually, the Vice-Rector for Scientific Affairs had this explanatory note forwarded to me (the previous one is here):
Meanwhile, thanks to the hint from my second whistleblower, the Polish Academy of science might be interested in checking these two publications by the above mentioned Gabryel collaborator Basta-Kaim and her PhD student Slusarczyk (full dossier here):
1. Ślusarczyk J, Trojan E, Glombik K, Piotrowska A, Budziszewska B, Kubera M, Popiolek-Barczyk K, Lason W, Mika J, Basta-Kaim A. Anti-inflammatory properties of tianeptine on lipopolysaccharide-induced changes in microglial cells involve toll-like receptor-related pathways. J Neurochem. 2016 Mar;136(5):958-70. doi: 10.1111/jnc.13452. PubMed PMID: 26640965.
Update 24.08.2017: this paper has been retracted on 11.07.2017 by the publisher Wiley, detailing all data manipulations. Quote from retraction notice:
“The Editorial Office was alerted by a science journalist that the same Western Blot lane had been used to represent two different proteins”.
2. Ślusarczyk J, Trojan E, Głombik K, Chamera K, Roman A, Budziszewska B, Basta-Kaim A. Fractalkine Attenuates Microglial Cell Activation Induced by Prenatal Stress. Neural Plast. 2016;2016:7258201. doi: 10.1155/2016/7258201. PubMed PMID: 27239349; PubMed Central PMCID: PMC4863104.
Update 18.01.2017. What does one say to this? A third Polish whistleblower contacted me with further evidence of apparent image re-use in these publications by Gabryel (full dossier here):
- Gabryel B, Adamczyk J, Huzarska M, Pudełko A, Trzeciak HI. Aniracetam attenuates apoptosis of astrocytes subjected to simulated ischemia in vitro. Neurotoxicology. 2002 Sep;23(3):385-95.
- Gabryel B, Pudelko A, Malecki A. Erk1/2 and Akt kinases are involved in the protective effect of aniracetam in astrocytes subjected to simulated ischemia in vitro. Eur J Pharmacol. 2004 Jun 28;494(2-3):111-20.
- Gabryel B, Chalimoniuk M, Stolecka A, Waniek K, Langfort J, Malecki A. Inhibition of arachidonic acid release by cytosolic phospholipase A2 is involved in the antiapoptotic effect of FK506 and cyclosporin a on astrocytes exposed to simulated ischemia in vitro. J Pharmacol Sci. 2006 Sep;102(1):77-87.
Now it’s not only western blots, but also microscopy images. How will the medical University of Silesia act upon this evidence?
Update 7.02.2017. The Vice Rector for Scientific Affairs of the Medical University of Silesia, Tomasz Szczepanski, notified me today (full letter here):
In reply to your e-mail dated on 18.01.2017 which contains new information about suspected falsification of research results in the mentioned publications I kindly inform that contained data has been sent to the competent an authority in this case“.
I can’t recall sending an email on that day to the university, but the update above is dated 18.01.2017. It’s good to know my reporting is being taken seriously at Katowice.
Bozena Gabryel was now elected to the Polish Academy of Science. as member of the Scientific Committee for the 2020-2023 term. Who says crime doesn’t pay?
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Some time back, said University started a postgraduate degree program on homeopathy
It was also reported on reddit (for those of you with only a Google Translational understanding of Polish)
It is no longer part of the curriculum
Here’s what the headmaster thinks of the criticism
Here’s at least one EU employee who put having attended the course on her CV
Click to access tomaszewskak_CV.pdf
Finally, the homeopathic studies program was ended
See also https://pl.wikipedia.org/wiki/Wyrok_SN_III_CSK_73/07_Boiron_S.A._vs._Gregosiewicz for a Polish court’s handling of a case where a professor’s criticism of a French homeopathic company was deemed to be in violation of the company’s personal rights (get that? We don’t either)
Polish Newsweek reports on Doctors Against Homeopathy
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With sadness I became acquainted with your document which values the results of our research in an unreliable way. I am surprised and it is incomprehensible why having doubts before the publication of our research neither you, Sir, or the two anonymous informants didn’t contact me about this case. The published article is harmful especially for my Phd student, who is just starting scientific career.
I would like to inform you that my participation as a co-author in one paper published by Dr. Gabryel was only restricted to discussion of the conception of this manuscript and I have not been involved in the experimental part. Western blot technique has been commonly used in our department by many researchers for almost 30 years. Each research is repeated many times. The samples are applied on the gel in the repetitions and combinations, and the results are calculated to the research from the many experiments. In the manuscripts we post a chosen representative blot. Recently for the research we use mainly ready-made gels (like Bio-rad) what allows us to obtain even striations and in many cases they are similar to each other, which is a contrary to the electrophoresis on gels prepared by us.
I inform you that all the membrane pictures included in the our papers (even the ones made few years back) have been automatically archived in the computer program, and their full documentation is available at any time to verify it, so if required the additional explanations remain available. Moreover after re-analysis the documentation of both papers I found the suggestion put forward in your article as unfounded, as well as the presentation of our research team and institution unjust.
Therefore I believe that as science journalist, the purpose of who is to promote reproductibility, honesty and fairness in academic research, you put rectification of article, which will restore the prestige of research and results obtained in our team.
Thank you for your comment. I guess you would rather not share those original western blot images? Could you also explain in more detail, how ready-made gels are able to generate bands which look that remarkably similar? I am genuinely curious.
Thus, you essentially deny everything and ask people to take back all their suspicions and to me, convincing proof, presented above. That’s not very mature. The argument that pre-casted gels will generate identical bands is childish. Computers can compare pixel patterns, you know that?
Fair point – so put the data up on FigShare or somewhere similar, so the community can see the that there are not issues. Without evidence, it is just hand waving.
As Richard Smith once said: When you see a fraudulent behaviour in a peer reviewed publication, don’t think even for a second that this is the first time that such person engages in something fraudulent, this is usually part of pattern, part of a career an a life that are fraudulent…
A correction appeared for Łabuzek et al, “Metformin increases phagocytosis and acidifies lysosomal/endosomal compartments in AMPK-dependent manner in rat primary microglia”. Naunyn Schmiedebergs Arch Pharmacol. 2010:
As you expected, there are further correction for papers. Strangely it appeared on the same day as previous one, and strangely there is exactly the same explanation
Erratum to: Ambivalent effects of compound C (dorsomorphin) on inflammatory response in LPS-stimulated rat primary microglial cultures
The authors regret that the Figure 6 containing representative western blots is not of sufficient quality and should be replaced. In the final steps of figure preparation the graphics technician made an incorrect insertion of blots into figure 6, which probably resulted from relatively low resolution of images.
In order to meet high standard of Naunyn-Schmiedebergs Archives of Pharmacology and to improve clarity of data presentation we wish to perform a corrigendum of figure 6, which would only affect graphical representation of WB bands and not scientific merit.
The authors would like to apologise for any inconvenience caused.
I hope it’s not my fault. The EiC of the journal, Roland Seifert, is professor at Hannover Medical School (MHH) which press office declared me persona non grata because of my reporting and won’t even tell me the time of the day 😉
And than authors (or corresponding author only?) publish a curious corrigendum:
Some editors just don’t care…
You might want to check this also https://pubpeer.com/publications/5615964D918F67D2B86CCE72F12E67#undefined
Thanks, I know the story. Australia-based researcher Gaetan Burgio and his lab did great work disproving NgAgo DNA editing claims. Not that he got any credit or even mention when Retraction Watch brought the news.
Sorry, I mean you should check the new comment posted by pubpeer. Then you will absolutely get a new picture of the “famous” author.
ah, I see. https://pubpeer.com/publications/5615964D918F67D2B86CCE72F12E67#14
Well, that supports my view that retractions due to “honest error” are extremely rare and should be put in museums.
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Dr.(?) Labuzek has his own TV show in Poland … A big ego, it seems.