Torturing Small Animals

As we are repeatedly educated, animal experiments are absolutely necessary to find cures for incurable human diseases. OK, this very well may be the theory, but what about the rather widespread practice?

Why do so many scientists keep subjecting animals to painful and cruel experiments to prove a fancy pet theory, which quite often fails to be proven experimentally, regardless of how many animals are being “used up”, ad these is why these same scientists resort to data forgery? So they can publish their fancy pet theory in a fancy journal and use it to celebrate themselves on the media and demand more money and more freedom to conduct even more painful and cruel animal experiments?

And to top it off: the “authorities” who are supposed to be preventing and sanctioning this animal abuse too often just don’t care. Let me give you some examples of what goes on.

Some months ago, Clare Francis reported this paper in a Wiley-published society journal:

Rosa Quiles-Perez , José Antonio Muñoz-Gámez , Ángeles Ruiz-Extremera , Francisco O’Valle , Laura Sanjuán-Nuñez , Ana Belén Martín-Alvarez , David Martín-Oliva , Trinidad Caballero , Paloma Muñoz De Rueda , Josefa León , Raúl Gonzalez , Jordi Muntané , Francisco Javier Oliver , Javier Salmerón Inhibition of poly adenosine diphosphate-ribose polymerase decreases hepatocellular carcinoma growth by modulation of tumor-related gene expression Hepatology (2010) doi: 10.1002/hep.23249

The mouse tumour xenografts are just too big, the animals had to suffer immensely and unnecessary. In USA, such experiments are regulated by Institutional Animal Care and Use Committees (IACUC), in this case specifically the guidelines say:

” Maximum tumor volume should not exceed 2000 (mm)ᶾ for mice”

The tumours were over 3000 mm³ big, thus 50% more than allowed. But this is how the journal’s Editor-in-Chief Gregory J. Gores, professor of medicine at Mayo Clinic and Executive Dean for Research there, replied to Clare Francis:

“We will review as per your request
But why do you assume the tumor volume is too big”

Because of IACUC guidelines maybe? Gores is actually responsible for their implementation, not just in his own lab but, in his capacity as executive dean for research, at the entire Mayo Clinic. And he pretends to be utterly clueless. Now imagine what kind of experiments must be going on at Mayo without anyone batting an eye at the excessive animal abuse.

I wrote to Gores about this, with Mayo’s IACUC officer in cc. There was of course no reply.

It wasn’t just tortured mice, but also fake data. That paper’s co-author, the Spanish liver researcher Jordi Muntane, has a hefty PubPeer record of falsified data in his papers, while promising to serve “in the care of patients with liver cancer“. Also the Figure 8A in his Hepatology paper was clearly forged:

Guess what message Gores had for Clare Francis?

“For the sake of due diligence, the figures were subjected to recent forensic analysis. No concerns were identified. Regarding Figure 1A specifically, we do not have sufficient concerns to warrant further pursuit of the issue. We now consider this issue resolved.“

Indeed. Animal abuse is bad enough, but scientists who don’t care about this part of research ethics, often care even less about another part: data manipulation. And people like Gores who are supposed to watch over both not only don’t give a flying toss, they even educate you that this exactly the right way to do science.

Here a typical Muntane paper by the way, with its fake gels:

Dalia Fouad , Emilio Siendones , Guadalupe Costán , Jordi Muntané Role of NF‐κB activation and nitric oxide expression during PGE ₁ protection against d ‐galactosamine‐induced cell death in cultured rat hepatocytes Liver international (2004) doi: 10.1111/j.1478-3231.2004.00913.x

This one is nice also:

Raúl González , Adolfo Cruz , Gustavo Ferrín , Pedro López-Cillero , Rubén Fernández-Rodríguez , Javier Briceño , Miguel A. Gómez , Sebastián Rufián , Manuel De La Mata , Antonio Martínez-Ruiz , Jose J.G. Marin , Jordi Muntané Nitric oxide mimics transcriptional and post-translational regulation during α-tocopherol cytoprotection against glycochenodeoxycholate-induced cell death in hepatocytes Journal of Hepatology (2011) doi: 10.1016/j.jhep.2010.10.022

Here another one by Muntane, I recognise other known cheaters among the authors, Javier Gonzalez-Gallego and his mentee Jose Mauriz:

María A. Rodríguez-Hernández , Raúl González, Ángel J. De La Rosa , Paloma Gallego , Raquel Ordóñez , Elena Navarro-Villarán, Laura Contreras, Mario Rodríguez-Arribas , Javier González-Gallego , José M. Álamo-Martínez , Luís M. Marín-Gómez , José A. Del Campo , José L. Quiles , José M. Fuentes, Jesús De La Cruz, José L. Mauriz , Francisco J. Padillo , Jordi Muntané Molecular characterization of autophagic and apoptotic signaling induced by sorafenib in liver cancer cells Journal of Cellular Physiology (2018) doi: 10.1002/jcp.26855

Muntane reacted by sharing raw data on PubPeer which soon proved to look, well, modified.

Unsurprisingly, Muntane also collaborates with the fraudster team around Angela Valverde in Madrid, together they publish papers with forged figures.

But enough of Muntane and his friends. In Spain research fraud and animal abuse are not just tolerated, those are actively celebrated. Recall how Carlos Lopez-Otin killed over 5000 “transgenic” mice to cover up tracks of research fraud, saluted by the Spanish scientific and political elites.

And then again, what can you expect from academic elites like Gores, right:

Yuko Akazawa , Justin L. Mott , Steven F. Bronk , Nathan W. Werneburg , Alisan Kahraman , Maria Eugenia Guicciardi , Xue Wei Meng , Shigeru Kohno , Vijay H. Shah , Scott H. Kaufmann , Mark A. McNiven , Gregory J. Gores Death receptor 5 internalization is required for lysosomal permeabilization by TRAIL in malignant liver cell lines Gastroenterology (2009) doi: 10.1053/j.gastro.2009.02.071

Another example of science by Gores, corrected in May 2022, with the entire notice consisting of “In Figure 4A the Bcl-x_L 231 panel was incorrect“:

Animal abuse and research fraud often go hand-in-hand (and bullying, and other forms of abuse for that matter, but that is another story). Let’s recall the former star cancer and ageing researcher in Germany, Karl Lenhard Rudolph, whose Fritz-Lippmann-Institute (FLI) in Jena first faced police raids for “on suspected breaches of the animal welfare and drug law, as well as embezzlement”, and then Rudolph was slapped with research misconduct findings by the DFG and the Leibniz society and forced to resign as FLI director. Yet despite proven data falsification, none of the misconduct-tainted papers were retracted. Because Rudolph seems to have made a deal with the journal editors.

Across the ocean, the Harvard professor Sam W Lee was initially fingered for gross mouse abuse.

Lakshmi Raj , Takao Ide , Aditi U. Gurkar , Michael Foley , Monica Schenone , Xiaoyu Li , Nicola J. Tolliday , Todd R. Golub , Steven A. Carr , Alykhan F. Shamji , Andrew M. Stern , Anna Mandinova , Stuart L. Schreiber , Sam W. Lee Selective killing of cancer cells by a small molecule targeting the stress response to ROS Nature (2011) doi: 10.1038/nature10167 

So how did Nature resolve this initially? With a huge Correction in 2012, where some rigged data was quietly fixed and where some of the evidence of animal torture was simply removed.

“We have also been unable to verify without doubt that the image in Supplementary Fig. 9b shows four different mice within the treated and untreated groups and therefore wish to replace this figure (see Fig. 2 of this Corrigendum).”

Look what Nature admitted as replacement:

This is pure sadism. In 2015, Nature had to issue another big Correction to purge the evidence once again:

“In this Letter, we presented findings from experiments using the EJ bladder xenograft cancer model, in which some tumours on some of the animals exceeded the maximum size (15 mm in any dimension) permitted by the Institutional Animal Care and Use Committee (IACUC) at Massachusetts General Hospital (MGH). Therefore, we withdraw the data presented in Supplementary Fig. 9b, as well as in Fig. 2 from the first Corrigendum. Although other tumours were found to exceed the permitted maximum, owing to the degree of tumour size and the circumstances of the experimental procedures, this was considered acceptable…”

Maybe I should mention that Lee trained as postdoc at Mount Sinai under Stuart Aaronson, a toxic bully with his own research integrity problems:

It is quite likely Aaronson pulled strings to protect his mentee. It didn’t help in the long run, Lee’s fraud was just too massive. In 2018 and after two Corrections, his Nature paper was retracted, but not for animal abuse, but for data forgery:

“This Letter is being retracted owing to issues with Fig. 1d and Supplementary Fig. 31b, and the unavailability of original data for these figures, which raises concerns regarding the integrity of the figures. Nature published two previous corrections related to this Letter^1,2. These issues in aggregate undermine the confidence in the integrity of this study.”

At that time, Lee already had two retractions for fraud, and two more followed, making it five. His PubPeer record has over 30 fraudulent papers. Here is one with an eternal Expression of Concern, where Lee personally is blamed:

Sam W. Lee , Corinne L. Reimer , Li Fang , M. Luisa Iruela-Arispe , Stuart A. Aaronson Overexpression of kinase-associated phosphatase (KAP) in breast and prostate cancer and inhibition of the transformed phenotype by antisense KAP expression Molecular and Cellular Biology (2000) doi: 10.1128/mcb.20.5.1723-1732.2000 

In 2021, Lee settled an agreement with the US Department of Justice, which issued this press release:

“Sam W. Lee, PhD, 67, of Bellevue, Wash., has agreed to pay $215,000 to resolve allegations that he submitted false claims for payment in a grant application to the NIH. MGH, the sponsor institution for the NIH grant, disclosed to the United States that Dr. Lee, the Principal Investigator (PI), submitted the grant application to NIH containing allegedly inauthentic data. MGH separately repaid NIH the full amount of funds it drew from the grant. […]

The United States contends that Dr. Lee altered the experiment descriptions in two of the figures, falsifying the results of the experiments, and that Dr. Lee falsified a third figure by horizontally flipping the image and thus mislabeling the results in the application.”

Torturing animals for fake science is an ever-green classic. Recall the Sicilian fraudster team around Francesco Squadrito:

The journals and even the Italian authorities can’t care less.

Ok, let’s swoop back to USA and meet the German professor at Stony Brook Cancer Center in New York, Ute Moll. Who refused all communication when faced with PubPeer evidence of both animal abuse and data manipulation in her papers, because she is busy with “changing the lives of colorectal cancer patients“.

#StonyBrookU prof Ute Moll's #research in genetic manipulation is changing the lives of colorectal cancer patients by markedly inhibiting tumor growth and invasiveness. https://t.co/nFKGmn3Uww @theNCI @StonyBrookMed #Innovation #Momentum #TuesdayThoughts

— Stony Brook University (@stonybrooku) September 18, 2018

This paper is just 2 years old, there is no lame excuse of times having been different. Basically, the mice were tortured in the Moll lab while elsewhere at Stony Brook students received courses on animal protection regulations:

Karis Tutuska , Laura Parrilla-Monge , Erica Di Cesare , Alice Nemajerova , Ute M. Moll Statin as anti-cancer therapy in autochthonous T-lymphomas expressing stabilized gain-of-function mutant p53 proteins Cell Death & Disease (2020) doi: 10.1038/s41419-020-2466-4 

Because Moll remained silent, I reported this paper and the ones that follow to the Assistant Vice President for Research Compliance at the Stony Brook University, who confirmed receiving my notification. I did not contact that Nature Publishing Group journal because its editors’ stance on research integrity is as credible as a bunch cats preaching veganism.

And here come the papers with manipulated data for Stony Brook to investigate:

Alice Nemajerova , Oleksi Petrenko , Lorenz Trümper , Gustavo Palacios , Ute M. Moll Loss of p73 promotes dissemination of Myc-induced B cell lymphomas in mice Journal of Clinical Investigation (2010) doi: 10.1172/jci40331 

A cloned gel lane. Obviously the mouse experiments didn’t work out as desired. Here an old cell culture study:

In case you are not sure if the gel bands are indeed cloned, what with the low resolution of that old figure, here is more by Moll’s mentee Alexander Zaika (now professor at the University of Miami, Sylvester Health Center):

Alexander Zaika , Natalia Marchenko , Ute M. Moll Cytoplasmically “Sequestered” Wild Type p53 Protein Is Resistant to Mdm2-mediated Degradation Journal of Biological Chemistry (1999) doi: 10.1074/jbc.274.39.27474

The Miami professor remained silent when I asked him to comment on his papers with Moll.

Alex I. Zaika , Neda Slade , Susan H. Erster , Christine Sansome , Troy W. Joseph , Michael Pearl , Eva Chalas , Ute M. Moll DeltaNp73, a dominant-negative inhibitor of wild-type p53 and TAp73, is up-regulated in human tumors The Journal of Experimental Medicine (2002) doi: 10.1084/jem.20020179

Hope you like this Mo-Zaika:

Oleksi Petrenko , Alexander Zaika , Ute M. Moll deltaNp73 facilitates cell immortalization and cooperates with oncogenic Ras in cellular transformation in vivo Molecular and Cellular Biology (2003) doi: 10.1128/mcb.23.16.5540-5555.2003 

Oleksi Petrenko is still under Moll’s wings, as a “research assistant professor” at Stony Brook.

Flaminia Talos , Oleksi Petrenko , Patricio Mena , Ute M. Moll Mitochondrially Targeted p53 Has Tumor Suppressor Activities In vivo Cancer Research (2005) doi: 10.1158/0008-5472.can-05-1084

This paper was flagged on PubPeer only 2 years after it was published. But given how Elsevier’s Cell Press (Fraud-R-Us) runs its business, they probably just are just waiting out to declare the paper is too old to be corrected.

Angelina V. Vaseva , Natalie D. Marchenko , Kyungmin Ji , Stella E. Tsirka , Sonja Holzmann , Ute M. Moll p53 opens the mitochondrial permeability transition pore to trigger necrosis Cell (2012) doi: 10.1016/j.cell.2012.05.014

Moll happens to be a guest professor at the University of Göttingen in Germany, which explains this collaboration:

Ramona Schulz , Natalia D. Marchenko , Lena Holembowski , Günter Fingerle-Rowson , Marina Pesic , Lars Zender , Matthias Dobbelstein , Ute M. Moll Inhibiting the HSP90 chaperone destabilizes macrophage migration inhibitory factor and thereby inhibits breast tumor progression The Journal of Experimental Medicine (2012) doi: 10.1084/jem.20111117 

The paper was already corrected for data inconsistencies in 2012:

“The authors regret that they incorrectly described the error bars as indicating the mean in several legends in the original manuscript. The error bars indicate the standard deviation. In addition, minor errors in panel letter order (Fig. 4) and a figure legend title (Fig. S1) were noted. These errors have been corrected in the html and pdf versions of this article.”

There are several reasons why another correction or even retraction of this paper is utterly out of the question.

• Reason 1: Moll is a big professor at Stony Brook, USA, and guest professor at the University of Göttingen, Germany
• Reason 2: Matthias Dobbelstein is a big professor and institute director at the University of Göttingen, Germany
• Reason 3: Lars Zender is a big professor and clinic director director at the University of Tübingen, Germany

This figure in another Göttingen collaboration and provides a clue how the work is done:

Daniela Kramer , Nadine Stark , Ramona Schulz-Heddergott , Norman Erytch , Shelley Edmunds , Laura Roßmann , Holger Bastians , Nicole Concin , Ute M Moll , Matthias Dobbelstein Strong antitumor synergy between DNA crosslinking and HSP90 inhibition causes massive premitotic DNA fragmentation in ovarian cancer cells Cell Death & Differentiation (2017) doi: 10.1038/cdd.2016.124

Basically, there are no proper loading controls because each panel used at least three gels: one (or two) gel(s)for Fanc protein analysis, one gel for BLM, and yet another gel for the loading control HSC70. We must trust the authors to have been honest and not having misloaded the gels. But you saw the other Moll-Dobbelstein paper above.

Rule of thumb is that most publishers will want to avoid the hustle and do little to nothing, either on research fraud or on animal abuse.

In fact, it turns out, that at Wiley ethics approvals for animal research are recommended, but not really mandatory. If authors get caught on having lied about those, it’s enough if they retrospectively auto-certify themselves, by assuring to have adhered to all the rules anyway. Not really that shocking knowing that Wiley is even OK with papermills.

An Erratum was issued for this paper in June 2021:

Lei Zhang, Fei Li, Wang-Jun Qin, Chao Fu, Xiang-Lin Zhang Changes in intestinal microbiota affect metabolism of ginsenoside Re Biomedical Chromatography (2021) https://doi.org/10.1002/bmc.5189

“In Section 2.2. ‘Animal Experiments’ of their article, the authors noted that: “All the animal studies were conducted in accordance with protocols approved by the Animal Ethics Committee of China Japan Friendship Hospital” (Zhang et al., 2018). Following concerns raised to the journal by a reader regarding the study’s ethical approval process, the authors would like to inform readers that, due to a translation error and a misunderstanding of the process, prospective ethical approval for this study was mistakenly not sought. However, the protocols followed for this study in relation to the care and use of animals were undertaken in accordance with the animal care and use policies of the Animal Ethical and Welfare Committee (AEWC) of the Clinical Institute of China Japan Friendship Hospital.

The authors apologise for this error.”

The more alarming case would be that from the 2 pre-teen daughters of Professor Hongliang Li, who completed an animal research on anti cancer benefits of green tea. The 2 elementary school kids performed tumor dissection from the animals, as they claimed and won national prize https://t.co/OJD80Iq6P4

— 虎仔 🇺🇦 (@TigerBB8) July 20, 2020

Tulipa Fosteriana quoted on PubPeer what kind of experiments the authors did, which are apparently “Classification D or above for Animal Pain and Distress Levels”. Highlights theirs:

“The OACS model was established by exposing the rats to continuous fatigue and acute cold stress. The specific steps were as follows: a weight(~5% of the body weight) was tied to the tail of each rat, and the rats were then put in a home made swimming pool. The rats swam twice daily at a 10 min interval for a total of 14 days. The water temperature was maintained at 25 ± 2°C on the first 13 days and at 0 ± 2°C on the last day. “

Thing is, the authors didn’t make a mistake while abusing rats without an ethics approval. They intentionally lied to get the paper published in that Wiley journal. Its Editor-in-Chief and University of Georgia professor Michael Bartlett was informed by Tiger BB8 already in September 2020:

“The first (Lei Zhang) and the corresponding (Xiang-Lin Zhang) authors are affiliated with China Japan Friendship Hospital in Beijing, China. In their paper, the authors stated that the animal protocol was approved by the IACUC of the hospital and they purchased their SD rats from Beijing University School of Medicine. However, based on the information I obtained from their colleagues (prefer to remain anonymous), the hospital recently received an allegation of misconduct and conducted an investigation on this paper. The hospital found that there was never such a protocol reviewed and approved by their IACUC and requested an explanation from the authors. The first author explained that the animal protocol was actually approved by the IACUC of University of Nebraska Medical Center (where the second author Fei Li and the 4^th author Chao Fu were) and the experiment was performed there when he was a visiting scholar for a year. When questioned further, Zhang said that the experimental materials stated in his paper as purchased in China were indeed purchased in China and he brought them over to UNMC while visiting there.

However, that story really doesn’t sound plausible. Based on my investigation, there is not faculty at College of Pharmacy of UNMC with a name as Fei Li or Chao Fu. Actually, Chao Fu was a PharmD student there from 2014-2018 and is a practicing pharmacist in Washington. As we all know, no student can be the PI of an animal protocol, per IACUC policy.“

So what with the corrigendum, it now seems to be official Wiley policy that ethics approvals for animal research are not long mandatory. The publisher’s executive editor Paul Trevorrow who approved that Erratum, promised to me to explain. That was in September 2021. Silence ever since, despite reminders.

Here is a crazy idea: maybe animal abuse should be a criminal offence, not just for private people, but also for those who do it professionally, i.e. meat industry and, imagine this, even for scientists? Here an even crazier idea: if a scientist falsified the results of animal experiments, all these experiments should be treated as animal abuse, even if the protocols were originally officially approved.

Yeah, an insane idea.

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