Say what you want about Chinese papermill fraud, but at least the rodent experiments they describe are entirely fictional. No animals were harmed in the production of those fake biomedical papers from China, but it is quite different with this Sicilian papermill which my colleagues Cheshire (Actinopolyspora biskrensis) and Aneurus Inconstans uncovered. Approved by the University of Messina, many rodents were subjected to cruel experiments, but instead of real scientific results, only Photoshop fraud was published. Virtually every “original research” paper which the sleuths looked at contained fake data. All because there was money to be made: the Fraud Squad patented some flavanol drugs (one later proved to be toxic), and they soon enough moved to experimenting on… children.
This looks like a gigantic research fraud scandal, even for Italian standards. It spans two decades and involves many people, mostly at the University of Messina. At the centre is the pharmacology professor Francesco Squadrito.
And it is not the first Sicilian fraud ring my colleagues uncovered. Here is another gang in Palermo, who managed to forge tobacco research even:
But this flavanol fraud affair in Messina seems to be much, much bigger. Let me introduce you to other key members of Squadrito’s Fraud Squad. First of all, there is the University of Messina pharmacology professor Domenica Altavilla, who also happens to be Squadrito’s wife. I found out these two are married from a Facebook post by the Rotary Club in Messina from 2017, which mentions:
There are more family members in the gang, as co-authors of fraudulent papers. There is another Messina professor named Giovanni Squadrito, presumably the ringleader’s brother. There seems to be other family relations involved, like Rolando Marini (deceased in 2019) and Herbert Ryan Marini (an US-American who possibly married Rolando’s daughter). The Messina professors Giuseppe Vita and Carmelo Romeo are likely the fathers of the Gian Luca Vita (local neurologist), and Sara Romeo (postdoc in UK), respectively.
In Italy, women keep their maiden name when marrying, so it is difficult to establish who is married to whom there. Yet, entire extended families are known to be employed at same Italian university.
The Messina professor Alessandra Bitto is a major figure in the affair, her name is on almost all of the fake papers I will present, but she doesn’t seem to be married to someone else, not a co-author of the fake papers. A younger Messina gang member with a big role is the assistant professor Natasha Irrera, she calls Squadrito her “scientific father“. Another important name is Salvatore Guarini, a Sicilian based at the University of Modena and Reggio Emilia in Northern Italy, Bitto did her PhD there under his supervision.
Finally, for all we know a regular member of the Fraud Squad, the University of Messina professor Letteria Minutoli, might be related to top governmental officials in the commune of Messina (Massimo Minutoli and Caterina Minutoli), which would preclude all attempts of investigation into the affair.
And here is Squadrito himself telling you about the curative magic of the bergamot fruit. The man is obsessed with plant flavanols.
Now, let us finally look at all these fake papers. We shall start with a Squadrito-Altavilla-Bitto-Irrera paper which was recently corrected by the publisher Wiley.
Alessandra Bitto, Domenica Altavilla , Gabriele Pizzino , Natasha Irrera , Giovanni Pallio , Michele R Colonna , Francesco Squadrito Inhibition of inflammasome activation improves the impaired pattern of healing in genetically diabetic mice British journal of pharmacology (2014) doi: 10.1111/bph.12557
This was the correction notice, issued in August 2021:
“Accidental mistakes during article preparation led to duplications in Figures 1b, d, 2a and 3a.
Since substantial time has elapsed since the study was originally conducted, the authors were not able to retrieve the original images. The authors have therefore re-probed the original protein samples and re-run the western blots with the antibodies currently in use.
Images of the re-run analyses are provided below. As per current BJP guidelines, the authors have set a single level of significance as P < 0.05. The statistical analysis shown is one way ANOVA with Tukey’s multiple comparisons post-tests conducted only if F was significant at (P < 0.05).
The authors sincerely apologize for these mistakes and state that these mistakes do not affect the quantification or conclusions of this study.”
Now look at the Figure 2A.
This PubPeer exchange shows that in the Squadrito and Altavilla labs “raw data” is a foreign concept. Anything can be raw data: unrelated gel images, digital fabrications, just label it any way you feel like. The journals approve. Well, anyway, MDPI does.
That corrected paper used diabetic mouse model to have their backs slashed to study wound healing. The team did a lot of animal research of this kind, all of its results were faked in Photoshop.
Squadrito doesn’t reply on PubPeer anymore. Initially, he did, here is one such case (a rare cell culture study without any abused rodents, btw):
Federica Mannino , Giovanni Pallio, Roberta Corsaro , Letteria Minutoli, Domenica Altavilla , Giovanna Vermiglio , Alessandro Allegra , Ali H. Eid , Alessandra Bitto, Francesco Squadrito , Natasha Irrera Beta-Caryophyllene Exhibits Anti-Proliferative Effects through Apoptosis Induction and Cell Cycle Modulation in Multiple Myeloma Cells Cancers (2021) doi: 10.3390/cancers13225741
Squadrito replied on PubPeer:
That supplemental file which MDPI published online was not really raw data. It looked as if someone scanned an empty X-Ray film and digitally pasted a string of gel bands onto it.
This kind of gel image is very similar to what the fraudsters of Cassava Science presented to the journals as “raw data”.
As Cheshire also noted, “The supplemental files provided on the journal site do not appear to include the control bands, although one would expect to see them on the same gel.” So Squadrito send MDPI another gel image and the publisher happily added that.
Yep, because the originally published actin gel obviously never existed as such, they just sent a random picture of some other western blot experiment. Who cares.
Nobody also cares about all that animal abuse which was absolutely unnecessary because the authors faked the results anyway.
In December 2014, someone (possibly a former lab member) shared something on PubPeer regarding a Squadrito gang paper, the posted link is broken by now. But in November 2016, this person returned to share their concerns as an image:
Letteria Minutoli, Alessandra Bitto, Francesco Squadrito, Natasha Irrera , Mariagrazia Rinaldi , Piero Antonio Nicotina , Salvatore Arena , Carlo Magno , Herbert Marini , Luca Spaccapelo , Alessandra Ottani , Daniela Giuliani , Carmelo Romeo , Salvatore Guarini , Pietro Antonuccio, Domenica Altavilla Melanocortin 4 receptor activation protects against testicular ischemia-reperfusion injury by triggering the cholinergic antiinflammatory pathway Endocrinology (2011) doi: 10.1210/en.2011-1016
What’s wrong here? Every one of these four actin western blots is a composite of one single gel band cloned 8 times.
Here, live rats had their testicles torn off:
“Rats were anesthetized with an ip injection of 50 mg/kg of pentobarbital sodium. Testicular ischemia-reperfusion (TI/R) injury was induced by torsion of the left testis, with a 720° twisting of the spermatic cord so as to produce a total occlusion of testis for 1 h. The same testis was then detorted.”
Note that the rats did not receive any anaesthetic after the testicle-twisting operation and suffered in pain for a whole day until they were killed.
More testicular torture on rats:
Pietro Antonuccio , Letteria Minutoli , Carmelo Romeo , Piero Antonio Nicòtina , Alessandra Bitto, Salvatore Arena , Domenica Altavilla, Biagio Zuccarello , Francesca Polito, Francesco Squadrito Lipid peroxidation activates mitogen-activated protein kinases in testicular ischemia-reperfusion injury The Journal of Urology (2006) doi: 10.1016/j.juro.2006.06.086
“All animals were anesthetized with an intraperitoneal injection of 50 mg/kg pentobarbital sodium. TI/R [Testicular Ischemia-Reperfusion Injury,- LS] was induced by torsion of the left testis with 720-degree twisting of the spermatic cord to produce total testicular occlusion for 1 hour. The same testis was then detorsed. Following 0, 10, 15,20, 25 and 30 minutes, and 1, 3 and 24 hours of reperfusion, respectively, rats were sacrificed with an overdose of pentobarbital sodium and bilateral orchiectomy was performed.”
All to produce this fake data:
Also these rats had their testicles ripped off, with these results:
Letteria Minutoli, Pietro Antonuccio, Francesca Polito, Alessandra Bitto , Francesco Squadrito, Natasha Irrera, Piero Antonio Nicotina , Carmine Fazzari , Angela Simona Montalto , Vincenzo Di Stefano , Carmelo Romeo, Domenica Altavilla Peroxisome proliferator activated receptor beta/delta activation prevents extracellular regulated kinase 1/2 phosphorylation and protects the testis from ischemia and reperfusion injury The Journal of Urology (2009) doi: 10.1016/j.juro.2008.11.095
More rats with their testicles torn off:
Letteria Minutoli , Pietro Antonuccio , Carmelo Romeo , Piero Antonio Nicòtina , Alessandra Bitto, Salvatore Arena , Francesca Polito , Domenica Altavilla , Nunzio Turiaco , Antonio Cutrupi , Biagio Zuccarello , Francesco Squadrito Evidence for a role of mitogen-activated protein kinase 3/mitogen-activated protein kinase in the development of testicular ischemia-reperfusion injury Biology of Reproduction (2005) doi: 10.1095/biolreprod.105.040741
Not just rats, also mice were subjected to testicle twisting:
Letteria Minutoli, Pietro Antonuccio , Francesca Polito, Alessandra Bitto, Tiziana Fiumara , Francesco Squadrito, Piero Antonio Nicotina , Salvatore Arena , Herbert Marini, Carmelo Romeo, Domenica Altavilla Involvement of mitogen-activated protein kinases (MAPKs) during testicular ischemia-reperfusion injury in nuclear factor-kappaB knock-out mice Life Sciences (2007) doi: 10.1016/j.lfs.2007.06.016
“Mice were anesthetized with an intraperitoneal injection of 80 mg/kg of pentobarbital sodium. Testis ischemia–reperfusion injury (TI/R) was induced by torsion of the left testis, with a 720° twisting of the spermatic cord so as to produce a total occlusion of testis for one hour. The same testis was then detorted. Following 0, 10, 30 min and 1, 3 and 24 h of reperfusion, the animals were sacrificed with an overdose of pentobarbital sodium and bilateral orchidectomies were performed.”
The Messina professors had their fun:
More testicle torture, mice again:
Letteria Minutoli , Pietro Antonuccio, Francesca Polito, Alessandra Bitto, Francesco Squadrito, Vincenzo Di Stefano , Piero Antonio Nicotina , Carmine Fazzari , Daniele Maisano, Carmelo Romeo, Domenica Altavilla Mitogen-activated protein kinase 3/mitogen-activated protein kinase 1 activates apoptosis during testicular ischemia-reperfusion injury in a nuclear factor-kappaB-independent manner European Journal of Pharmacology (2009) doi: 10.1016/j.ejphar.2008.12.028
“Animals were anesthetized with an intraperitoneal injection of 80 mg\kg of pentobarbital sodium to perform torsion of the left testis and spermatic cord, 720° for 1 h.”
They just kept tearing off rodents’ testicles, but what for? Because the results were always faked in Photoshop.
The team was tearing off testicles from live rodents for years, so much fun they had, “authorized by our local institution“:
Letteria Minutoli, Pietro Antonuccio , Natasha Irrera , Mariagrazia Rinaldi , Alessandra Bitto, Herbert Marini , Gabriele Pizzino , Carmelo Romeo , Antonina Pisani , Giuseppe Santoro , Domenico Puzzolo , Carlo Magno , Francesco Squadrito , Antonio Micali , Domenica Altavilla NLRP3 Inflammasome Involvement in the Organ Damage and Impaired Spermatogenesis Induced by Testicular Ischemia and Reperfusion in Mice The Journal of pharmacology and experimental therapeutics (2015) doi: 10.1124/jpet.115.226936
“Both WT and KO mice (total number = 126; each group consisted of seven animals) were anesthetized with an intraperitoneal injection of 80 mg/kg of pentobarbital sodium and torsion of the left testis and spermatic cord was performed as previously described (Minutoli et al., 2009). Then, the same testis was detorted.”
Everything in this paper is fake:
The latter image reuse involved this Frontiers paper with other problems (the editor was Squadrito’s own university rector, Salvatore Cuzzocrea). The mice were injected with the toxic heavy metal cadmium (Cd):
Francesco Squadrito, Antonio Micali , Mariagrazia Rinaldi , Natasha Irrera , Herbert Marini , Domenico Puzzolo , Antonina Pisani , Cesare Lorenzini , Andrea Valenti , Rosaria Laurà , Antonino Germanà , Alessandra Bitto, Gabriele Pizzino , Giovanni Pallio , Domenica Altavilla , Letteria Minutoli Polydeoxyribonucleotide, an Adenosine-A2A Receptor Agonist, Preserves Blood Testis Barrier from Cadmium-Induced Injury Frontiers in Pharmacology (2017) doi: 10.3389/fphar.2016.00537
In this paper, Cheshire noted:
“”The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. […]
However, the paper does not seem to mention that the first author notes on his CV states that he was “Principal Investigator 2007-2010 – Mastelli s.r.l” which seems to have invented and holds the patent on PDRN, which is the subject of this research. Perhaps related to this, the first author may also be a co-inventor [on a patent ](https://patents.google.com/patent/EP3377075A1/en?assignee=Mastelli+S.R.L.)currently assigned to Mastelli related to PDRN (that patent also lists Giulia Cattarini Mastelli and Laura Cattarini Mastelli as inventors).”
We will hear more of Squadrito’s COIs later. But for now you must understand: his own top boss, the University of Messina rector magnificus Cuzzocrea, obviously fully supports all this misconduct, even jumping in as editor when needed. This is Italian academia in all its pride, as we know it from Giorgio Zauli affair.
Anyway, some more testicular damage. Here, male rats received a surgery which caused varicocele (testicle vein enlargement), the “protocol was approved by the Committee of Animal Health and Care of University of Messina“:
Letteria Minutoli, Salvatore Arena , Pietro Antonuccio , Carmelo Romeo , Alessandra Bitto, Carlo Magno , Mariagrazia Rinaldi , Antonio Micali , Natasha Irrera , Gabriele Pizzino , Federica Galfo , Francesco Squadrito, Domenica Altavilla , Herbert Marini Role of Inhibitors of Apoptosis Proteins in Testicular Function and Male Fertility: Effects of Polydeoxyribonucleotide Administration in Experimental Varicocele BioMed research international (2015) doi: 10.1155/2015/248976
Too rpoduce these forgeries as results:
But now let’s see what other creative things Squadrito’s team did to rodents.
Bleed, poison and boil
Here, the mice were merely injected with drugs to cause prostata tumours, and not subjected to “surgeries”:
Letteria Minutoli, Domenica Altavilla , Herbert Marini, Mariagrazia Rinaldi , Natasha Irrera , Gabriele Pizzino , Alessandra Bitto , Salvatore Arena , Sebastiano Cimino , Francesco Squadrito, Giorgio Ivan Russo, Giuseppe Morgia Inhibitors of apoptosis proteins in experimental benign prostatic hyperplasia: effects of serenoa repens, selenium and lycopene Journal of Biomedical Science (2014) doi: 10.1186/1423-0127-21-19
But of course, there was no restraint in fraud. Look what the Figure 1 alone looks like:
All that was just Figure 1, mind you.
In the next study, a number of things was done to live rats, in particular very slowly bleeding them to death:
Salvatore Guarini , Domenica Altavilla, Maria-Michela Cainazzo , Daniela Giuliani , Albertino Bigiani , Herbert Marini , Giovanni Squadrito, Letteria Minutoli, Alfio Bertolini , Rolando Marini , Elena B. Adamo , Francesco S. Venuti , Francesco Squadrito Efferent vagal fibre stimulation blunts nuclear factor-kappaB activation and protects against hypovolemic hemorrhagic shock Circulation (2003) doi: 10.1161/01.cir.0000050627.90734.ed
“Under general anesthesia (urethane, 1.25 g/kg intraperitoneally) and after heparinization (heparin sodium, 600 IU/kg), rats were instrumented with indwelling polyethylene catheters in the left common carotid artery to record arterial blood pressure and into the right iliac vein to inject drugs and to bleed. […]
Acute hypovolemic hemorrhagic shock14–16 was induced by a graded withdrawal of blood (1.8 to 2 mL per 100 g body weight) until the MAP fell and stabilized at levels of 35 to 40 mm Hg. […]
Two minutes before the start of bleeding, rats were subjected to bilateral cervical vagotomy or sham surgical procedures. Direct stimulation of both caudal vagal trunks was carried out by means of bipolar platinum electrodes connected to a stimulator. […] Survival rate and survival time were evaluated for 180 minutes after the bleeding was discontinued.”
This is the kind of fake data these scientists produced, possibly while waiting for the rats to die:
Also in this study rats were slowly bled to death, and its fake data was flagged on PubPeer already in May 2015:
Salvatore Guarini , Maria Michela Cainazzo , Daniela Giuliani , Chiara Mioni , Domenica Altavilla , Herbert Marini , Albertino Bigiani , Valeria Ghiaroni , Maria Passaniti , Sheila Leone , Carla Bazzani , Achille P Caputi , Francesco Squadrito, Alfio Bertolini Adrenocorticotropin reverses hemorrhagic shock in anesthetized rats through the rapid activation of a vagal anti-inflammatory pathway Cardiovascular Research (2004) doi: 10.1016/j.cardiores.2004.03.029
“Acute hemorrhagic shock was induced by a stepwise (within 20–25 min) withdrawal of about 50% of the circulating blood…”
Here, mice were fed alcohol to cause liver damage:
Domenica Altavilla, Herbert Marini, Paolo Seminara , Giovanni Squadrito, Letteria Minutoli, Maria Passaniti , Alessandra Bitto, Gioacchino Calapai , Margherita Calò , Achille P. Caputi , Francesco Squadrito Protective effects of antioxidant raxofelast in alcohol-induced liver disease in mice Pharmacology (2005) doi: 10.1159/000082939
Squadrito’s antioxidants can cure anything, as the Photoshop forgery testifies:
This study was particularly nasty, mice were dumped into 80°C hot water:
Mariarosaria Galeano , Domenica Altavilla, Alessandra Bitto, Letteria Minutoli, Margherita Calò , Patrizia Lo Cascio , Francesca Polito , Giovanni Giugliano , Giovanni Squadrito, Chiara Mioni , Daniela Giuliani , Francesco S. Venuti , Francesco Squadrito Recombinant human erythropoietin improves angiogenesis and wound healing in experimental burn wounds* Critical care medicine (2006) doi: 10.1097/01.ccm.0000206468.18653.ec
“After general anesthesia with sodium pentobarbital (80 mg/kg), hair on the back was shaved […] Mice were inserted headfirst into a burn template so that the dorsum was centered by the 2 3 cm window. The shaved skin on the back was immersed in 80°C water for 10 secs to achieve a second-degree scald burn. We produced a deep-dermal second degree burn.”
The mice were tortured again for further experiments, all to obtain this fake data:
More mouse boiling:
Natasha Irrera , Alessandra Bitto, Gabriele Pizzino , Mario Vaccaro , Francesco Squadrito, Mariarosaria Galeano , Francesco Stagno D’Alcontres , Ferdinando Stagno D’Alcontres , Michele Buemi , Letteria Minutoli , Michele Rosario Colonna , Domenica Altavilla Epoetin Alpha and Epoetin Zeta: A Comparative Study on Stimulation of Angiogenesis and Wound Repair in an Experimental Model of Burn Injury BioMed research international (2015) doi: 10.1155/2015/968927
“The protocol was evaluated and accepted by the Ethics Committee of the University of Messina […] Skin injury was performed on the shaved back of mice: after general anaesthesia with sodium pentobarbital (80 mg/kg/i.p.), hair on the back was shaved using a depilatory cream to reduce any possible injury due to hair removal. Burn injury was produced with 80°C water, and mice were immersed for 10 seconds with a burn template so that the dorsum was exposed to hot water through a 2 × 3 cm window. In this way, we produced a deep-dermal second-degree burn; postburn sedation and analgesia were provided with diazepam (50 mg/L added to drinking water) for 7 days to alleviate pain.”
Do we believe they really gave mice pain medicine after boiling their backs? Because they didn’t do this in earlier study. And anyway, nothing these authors claim can be trusted:
How about stroke?
Francesco Squadrito , Letteria Minutoli, Maria Esposito , Alessandra Bitto, Herbert Marini , Paolo Seminara , Alessandra Crisafulli , Mariolina Passaniti , Elena Bianca Adamo , Rolando Marini , Salvatore Guarini, Domenica Altavilla Lipid peroxidation triggers both c-Jun N-terminal kinase (JNK) and extracellular-regulated kinase (ERK) activation and neointimal hyperplasia induced by cessation of blood flow in the mouse carotid artery Atherosclerosis (2005) doi: 10.1016/j.atherosclerosis.2004.10.013
“Mice were anesthetized by intraperitoneal injection with a solution of xylazine (5 mg/kg) and ketamine (80 mg/kg). The left common carotid artery was exposed through a small mid-line incision in the neck. The artery was completely ligated
just proximal to the carotid bifurcation to disrupt blood flow.”
The results were then fabricated:
More stroke, but this time with Mongolian gerbils!
Unlike with most of Squadrito’s animal studies in Messina, this one was supervised by Guarini and “approved by the Committee on Animal Health and Care of Modena and Reggio Emilia University“
Daniela Giuliani, Alessandra Ottani , Letteria Minutoli, Vincenzo Di Stefano , Maria Galantucci , Alessandra Bitto, Davide Zaffe , Domenica Altavilla, Annibale R. Botticelli , Francesco Squadrito, Salvatore Guarini Functional recovery after delayed treatment of ischemic stroke with melanocortins is associated with overexpression of the activity-dependent gene Zif268 Brain Behavior and Immunity (2009) doi: 10.1016/j.bbi.2009.03.009
“Transient global brain ischemia was induced by occluding with atraumatic clips both common carotid arteries for 10 min”
The gerbils were abused to produce this bizarrely fake data:
More abused gerbils in Modena:
Daniela Giuliani , Chiara Mioni , Domenica Altavilla , Sheila Leone , Carla Bazzani , Letteria Minutoli, Alessandra Bitto , Maria-Michela Cainazzo , Herbert Marini , Davide Zaffe , Annibale R. Botticelli , Roberto Pizzala , Monica Savio , Daniela Necchi , Helgi B. Schiöth , Alfio Bertolini , Francesco Squadrito, Salvatore Guarini Both early and delayed treatment with melanocortin 4 receptor-stimulating melanocortins produces neuroprotection in cerebral ischemia Endocrinology (2006) doi: 10.1210/en.2005-0692
Again stroke was induced:
“Transient global brain ischemia was induced, under general anesthesia with chloral hydrate (400 mg/kg ip; Sigma, St. Louis, MO), by occluding with atraumatic clips both common carotid arteries for 10 min…”
And this is how one fakes the result:
Let me show you one utter insanity of the Squadrito team. This paper should be used as teaching material to educate students of biomedicine of the utter failure of peer review and scholarly publishing in general. It was published by a SAGE journal with an impact factor of 6.7.
Alessandra Ottani, Daniela Giuliani, Chiara Mioni , Maria Galantucci , Letteria Minutoli, Alessandra Bitto, Domenica Altavilla, Davide Zaffe , Annibale R Botticelli , Francesco Squadrito, Salvatore Guarini Vagus nerve mediates the protective effects of melanocortins against cerebral and systemic damage after ischemic stroke Journal of Cerebral Blood Flow & Metabolism (2009) doi: 10.1038/jcbfm.2008.140
Here the authors caused stroke in rats by injecting endothelin-1 into their brains (and removing the vagus nerve). But now observe what the figures of the paper look like:
The rest, Figures 1 and 7, are just bar diagrams. Maybe you noticed that the figures are not just boring repetitions: all gel bands are cloned. Strings of cloned gel bands, in every single western blot panel. As Aneurus Inconstans wrote on PubPeer:
Once, Squadrito and his team decided to prove that abdominal infection from a ripped appendix can be cured with pumping CO2 into the peritoneal cavity.
Angela Simona Montalto , Alessandra Bitto, Letteria Minutoli , Pietro Impellizzeri , Gaetano Costa , Natasha Irrera , Gabriele Pizzino , Francesco Squadrito, Domenica Altavilla , Carmelo Romeo CO2Pneumoperitoneum Preservesβ-Arrestin 2 Content and Reduces High Mobility Group Box-1 (HMGB-1) Expression in an Animal Model of Peritonitis Oxidative Medicine and Cellular Longevity (2015) doi: 10.1155/2015/160568
“…rats were anaesthetized with ether, and a midline incision was made below the diaphragm to expose the cecum. The cecum was ligated at the colon juncture with a 2-0 silk ligature suture without interrupting intestinal continuity. The cecum was punctured twice with an 18-gauge needle. The cecum was returned to the abdomen, and the incision was closed in layers with a 2-0 silk ligature suture. Six hours following CLP, animals were randomly selected to undergo CO2 pneumoperitoneum (LS) at a pressure of 5–7 mmHg or laparotomy…”
The real therapy was Photoshop of course:
There are so many ways to cause intestinal pain to make rodents suffer.
Letteria Minutoli, Domenica Altavilla, Alessandra Bitto, Francesca Polito, Ersilia Bellocco, Giuseppina Laganà, Daniela Giuliani, Tiziana Fiumara , Salvatore Magazù, Pietro Ruggeri , Salvatore Guarini, Francesco Squadrito The disaccharide trehalose inhibits proinflammatory phenotype activation in macrophages and prevents mortality in experimental septic shock Shock (Augusta, Ga.) (2007) doi: 10.1097/01.shk.0000235092.76292.bc
Rats were tortured by inducing inflammation shock:
“Rats were anesthetized with ethyl ether and a cannula (PE 50) was inserted into the right jugular vein to allow intravenous injection and facilitate blood withdraw.
Twenty-four hours after surgery endotoxin shock was induced in animals by a single intravenous injection of S. enteritidis LPS (20 mg/kg; LD90 ).”
The rats had to die in pain so the authors could publish this shocking fraud:
Also here rats were injected with LPS to cause massive inflammation:
Alessandra Bitto, Letteria Minutoli, Maria Rosaria Galeano, Domenica Altavilla, Francesca Polito, Tiziana Fiumara , Margherita Calò, Patrizia Lo Cascio, Lorena Zentilin , Mauro Giacca, Francesco Squadrito Angiopoietin-1 gene transfer improves impaired wound healing in genetically diabetic mice without increasing VEGF expression Clinical Science (2008) doi: 10.1042/cs20070250
The figures showing the desired experimental results were faked. Some of the data was recycled from the Shock paper.
The shocking Shock paper had other uses:
Domenica Altavilla, Francesca Polito, Alessandra Bitto, Letteria Minutoli, Elisabetta Miraldi, Tiziana Fiumara , Marco Biagi, Herbert Marini, Daniela Giachetti, Mario Vaccaro, Francesco Squadrito Anti-Inflammatory Effects of the Methanol Extract of Sedum telephium ssp. maximum in Lipopolysaccharide- Stimulated Rat Peritoneal Macrophages Pharmacology (2008) doi: 10.1159/000157626
For some reason, the authors didn’t torture rats before killing them to obtain macrophages. Or maybe they did? After all, the paper doesn’t say if the rats were killed before “washing the abdominal cavity with RPMI 1640“. Anyway, here is the result:
The Messina slashers
Sometimes the rodents were cut. How about some fake diabetes research:
Mariarosaria Galeano , Domenica Altavilla, Domenico Cucinotta , Giuseppina T. Russo , Margherita Calò , Alessandra Bitto, Herbert Marini, Rolando Marini, Elena B. Adamo , Paolo Seminara , Letteria Minutoli, Valerio Torre , Francesco Squadrito Recombinant human erythropoietin stimulates angiogenesis and wound healing in the genetically diabetic mouse Diabetes (2004) doi: 10.2337/diabetes.53.9.2509
Transgenic models of diabetic mice had their backs slashed:
“After general anesthesia with ketamine hydrochloride (110 mg/kg), the hair on the back was shaved and the skin washed with povidone-iodine solution and wiped with sterile water. Two full-thickness longitudinal incisions (4 cm) were made on the dorsum of the mice, and the wound edges were closed with skin clips placed at 1-cm intervals. Seven animals for each group were killed after 3, 6, and 12 days”
All to produce some fake immunohistochemistry:
Once again, diabetic mice were used and then “Two full-thickness longitudinal incisions (4 cm) were made on the dorsum of the mice“:
Mariarosaria Galeano, Francesca Polito, Alessandra Bitto, Natasha Irrera, Giuseppe M. Campo , Angela Avenoso , Margherita Calò, Patrizia Lo Cascio, Letteria Minutoli, Mauro Barone , Francesco Squadrito, Domenica Altavilla Systemic administration of high-molecular weight hyaluronan stimulates wound healing in genetically diabetic mice Biochimica et Biophysica Acta (2011) doi: 10.1016/j.bbadis.2011.03.012
To produce these completely made-up results:
More back-slashing of mice:
Letteria Minutoli, Domenica Altavilla, Alessandra Bitto, Francesca Polito, Ersilia Bellocco, Giuseppina Laganà, Tiziana Fiumara , Salvatore Magazù, Federica Migliardo , Francesco Saverio Venuti , Francesco Squadrito Trehalose: a biophysics approach to modulate the inflammatory response during endotoxic shock European Journal of Pharmacology (2008) doi: 10.1016/j.ejphar.2008.04.005
And then the results were faked:
That other paper (again with diabetic mice whose backs were slashed) reused immunohistochemistry images in other ways:
Alessandra Bitto, Letteria Minutoli, Domenica Altavilla, Francesca Polito, Tiziana Fiumara , Herbert Marini, Mariarosaria Galeano, Margherita Calò, Patrizia Lo Cascio, Michele Bonaiuto , Alba Migliorato, Achille P Caputi , Francesco Squadrito Simvastatin enhances VEGF production and ameliorates impaired wound healing in experimental diabetes Pharmacological Research (2008) doi: 10.1016/j.phrs.2008.01.005
More diabetic mice slashed:
Alessandra Bitto, Natasha Irrera, Letteria Minutoli, Margherita Calò, Patrizia Lo Cascio, Paolo Caccia, Gabriele Pizzino, Giovanni Pallio, Antonio Micali, Mario Vaccaro, Antonino Saitta, Francesco Squadrito, Domenica Altavilla Relaxin improves multiple markers of wound healing and ameliorates the disturbed healing pattern of genetically diabetic mice Clinical Science (2013) doi: 10.1042/cs20130105
“After general anaesthesia with sodium pentobarbital (80 mg/kg of body weight, intraperitonealy), hair on the back was shaved and two parallel 4-cm incisions were produced with the use of a scalpel (Figure 1A), on the back of all mice as described previously .”
As described previously, the results were fabricated:
More wound healing research, where mouse backs were slashed and the results faked in Photoshop:
Mariarosaria Galeano, Francesca Polito, Alessandra Bitto, Natasha Irrera, Giuseppe M. Campo , Angela Avenoso , Margherita Calò, Patrizia Lo Cascio, Letteria Minutoli, Mauro Barone , Francesco Squadrito, Domenica Altavilla Systemic administration of high-molecular weight hyaluronan stimulates wound healing in genetically diabetic mice Biochimica et Biophysica Acta (2011)
Other creative ways to torture animals while faking the results:
Alessandra Bitto, Francesca Polito, Domenica Altavilla, Letteria Minutoli, Alba Migliorato, Francesco Squadrito Polydeoxyribonucleotide (PDRN) restores blood flow in an experimental model of peripheral artery occlusive disease Journal of Vascular Surgery (2008) doi: 10.1016/j.jvs.2008.06.041
Rats were abused by cutting off blood flow to their hindlimbs:
“The left femoral artery was exposed aseptically through a 2-mm incision and isolated from the femoral vein and nerve, with care taken to avoid damage to vessels or nerve, then was ligated with 7-0 suture just distal to the bifurcation of the anterior epigastric and lateral caudal femoral arteries. Finally, the femoral artery was excised from the ligation to the point distally where it bifurcates into the saphenous and popliteal arteries. As a consequence, blood flow to the ischemic limb becomes completely dependent on the collateral vessels (HLI animals).”
The experimental outcome was forged:
Falsified Laser Doppler perfusion imaging data, one doesn’t see this every day. How exotic! And yes, there were also fake western blots:
Here, the team induced skin and lung fibrosis in mice, because why not.
This paper features a father and son team from Messina, Gianfilippo Bagnato (deceased in 2019) and Gianluca Bagnato, which makes me wonder why Bitto’s old mentor, the Messina professor Antonino Saitta didn’t also invite his son Carlo Saitta to join, as on his other papers with Bitto? Oh, and the Messina rheumatologist Marco Atteritano is quite possibly Bitto’s partner, judging from this photo.
Gianluca Bagnato , Alessandra Bitto , Natasha Irrera, Gabriele Pizzino , Donatella Sangari , Maurizio Cinquegrani , William Roberts , Marco Atteritano , Domenica Altavilla, Francesco Squadrito, Gianfilippo Bagnato , Antonino Saitta Propylthiouracil prevents cutaneous and pulmonary fibrosis in the reactive oxygen species murine model of systemic sclerosis Arthritis Research & Therapy (2013) doi: 10.1186/ar4300
“A total of 100 μl of solution containing HOCl was injected s.c. into the back of the mice, by using a 27-gauge needle, every day for 6 weeks. Mice (n = 10) from the HOCl group (n = 20) were randomly chosen to be treated with propylthiouracil
(Sigma-Aldrich, Italy///) at the dose of 12 mg/kg/day.
[…] PTU administration was initiated 30 minutes after the HOCl subcutaneous injection, and continued for 6 weeks.”
To obtain these fake figures:
How about hitting mice on the head with a hammer?
Natasha Irrera , Gabriele Pizzino , Margherita Calò , Giovanni Pallio , Federica Mannino , Fausto Famà , Vincenzo Arcoraci , Vincenzo Fodale , Antonio David , Cosentino Francesca , Letteria Minutoli, Emanuela Mazzon , Placido Bramanti , Francesco Squadrito, Domenica Altavilla , Alessandra Bitto Lack of the Nlrp3 Inflammasome Improves Mice Recovery Following Traumatic Brain Injury Frontiers in pharmacology (2017) doi: 10.3389/fphar.2017.00459
“…animals were anesthetized by intraperitoneal injection of ketamine/xylazine (80 and 10 mg/kg, respectively) and subjected to an impact-acceleration model of diffuse TBI, based on the model described by Marmarou et al. (Marmarou et al., 1994), with some modifications. Briefly, an incision was made along the sagittal midline to expose the skull and a 3 mm thick steel disk was placed on between λ and bregma sutures. Animals were arranged on a 10-cm foam bed to soften the impact. Briefly, a 8 g weight with a 5 mm diameter and 5.5 cm length was dropped from a distance of 1.27 m, to produce TBI. After TBI, the steel disk was removed and the skin immediately sutured; a 2% lidocaine jelly was spread on to the impact site to reduce pain.”
Why yes, the brain injury was real, “approved by the Ethic Committee of the University of Messina“, but the figures in this Frontiers paper fake.
Hammering live rodents on the heads (after scalping!) is a cherished tradition in Messina, in this earlier paper they used rats:
Alessandra Bitto, Francesca Polito , Natasha Irrera , Margherita Calò , Luca Spaccapelo , Herbert R. Marini , Daniela Giuliani , Alessandra Ottani , Mariagrazia Rinaldi , Letteria Minutoli , Salvatore Guarini , Francesco Squadrito, Domenica Altavilla Protective effects of melanocortins on short-term changes in a rat model of traumatic brain injury* Critical Care Medicine (2012) doi: 10.1097/ccm.0b013e318236efde
”After 1 wk of acclimatization, animals were anesthetized by intraperitoneal injection of sodium pentobarbital (50 mg/kg) and subjected to the impact-acceleration model of diffuse TBI, as previously described (12). Briefly, a 450-g weight was dropped from a distance of 2 m onto a 10-mm diameter by 3-mm thick steel disc that had been fixed in place on the exposed dorsal surface of the skull midway between and bregma. The animal was supported on a 10-cm foam bed that provided the deceleration after impact. After injury, the steel disc was removed and the skin sutured before termination of anesthesia. ”
“Again the actin control in 2B and 2D (red box) was used three years before in Bitto et al. 2010 J Vasc Surg […] where the control was for human carotid arteries samples, while here is supposed to be for rat brain tissue. Then bands duplications are present also within blots of TNF-a, Caspase 3 and BAX (green, magenta and turquoise boxes). Finally the same actin control in 3C was reused from Figure 2A of again Bitto et al. 2010 J Vasc Surg, where the control was for human carotid arteries samples, while here is supposed to be for rat brain tissue (yellow boxes).“
These people are trolling, no? But no limits for creative ways of torturing a small animal:
Sonia Messina, Domenica Altavilla, M’hammed Aguennouz , Paolo Seminara , Letteria Minutoli , Maria C. Monici , Alessandra Bitto , Anna Mazzeo, Herbert Marini, Francesco Squadrito, Giuseppe Vita Lipid peroxidation inhibition blunts nuclear factor-kappaB activation, reduces skeletal muscle degeneration, and enhances muscle function in mdx mice The American journal of pathology (2006) doi: 10.2353/ajpath.2006.050673
“Five-week-old mdx and WT mice have been treated for 5 weeks with intraperitoneal injections with either IRFI 042 (n:8; 20 mg/kg three times a week) or vehicle (n:8; dimethyl sulfoxide/NaCl 0.9%; 0.1:1 v/v; 0.2 mg/kg three times a week). At the end of the experiments, animals were anesthetized with an intraperitoneal administration of sodium pentobarbital (80 mg/kg). Then, blood, collected by intracardiac puncture, was drawn to analyze creatine kinase (CK) levels and the biceps, quadriceps, and extensor digitorum longus (EDL) muscles were removed bilaterally”
Let’s hope the injected anaesthetics were not as fake as these gels, because the mice were literally vivisected, alive:
The last author, the University of Messina professor Giuseppe Vita pleaded on PubPeer that some immigrant of Muslim origin (his Messina colleague professor Mhammed Aguennouz) was to blame for this horrid conspiracy against Italian science:
Shall we blame Aguennouz also for papers were he is not even co-author? Those foreigners, eh, Dr Vita?
The tech did it!
Aneurus Inconstans and Cheshire have found fraud in almost every Squadrito paper they screened, and, at press time, they have over a hundred papers more to look at. The Messina lab puts every Chinese papermill to shame, also because Chinese forgeries are of much better quality and not just a string of one gel band cloned 10 times over.
But then again, the Chinese forgers have to work harder, because scientific journals do not offer same trust and service to non-white authors. And especially certain kind of Italian “scientists” knows how to organise a scam ring and publish many papers.
Squadrito replied to me only once, and he blames not his faculty colleague Aguennouz, but a technician. This is what Squadrito wrote to me on 24 April 2022:
“Dear Leonid Schneider,
Thank you for your mail. We are providing explanation for the concerns regarding the western blot analysis of the papers you mentioned, but this is a procedure that need time. We are checking all the records, and I kindly ask you 3 working weeks and I will come back to you and PubPeer with proper explanation. At this time I wish to underline that the scientific meaning of the papers under inquire has been confirmed several times by other authors. The concerns are only related to the the western blot analysis. As you know the majority of the articles date back to 2005 (17 years ago !!!!) and the experiments have been carried out even earlier (18-19 years ago) . At that time a lab technician was in charge for the protein extraction, western blot analysis, calculations and graph preparation based on coded samples prepared by the researchers. She retired 5 years ago and we are not keeping the old files of published papers for longer than 5 years; moreover since 2014 we’ve got a new image acquisition system (the Li-Cor C-Digit), while years ago we used fixer and developer and then the image was acquired and processed. Working on coded samples many times the images needed to be cut to get in the proper order the samples, but as said before it was a specific task of our technician. Following the imput of PubPeer we are searching for the old protein samples and if we find the residual extracts in the -80° freezer we will run again the experiments. Finally I wish to underline tha nobody has been investigated for research misconduct
Again we apologize for this unpleasant situation and remain at your complete disposal for any further clarification
Francesco Squadrito “
It is not clear whom Squadrito and his gang will blame for the papers published in the last 5 years. Maybe another technician. Or Aguennouz.
It is also not clear if Squadrito will ever have to retract a single paper, despite all of them being entirely fictional (expect for the reality of abused animals). His networks involve even the rector of his university, who surely will eagerly attest to every inquiring journal that all results are reproducible and no conclusions were ever affected.
No other member of the Squadrito gang replied to my emails. And neither did his university, but again, what with the rector Cuzzocrea, what can one expect. That’s how it’s usually done in Italian universities.
But now let’s talk about Squadrito’s business activities.
Conflicts of Interest
It is all about plant substances with alleged anti-oxydative properties, specifically the flavanols genistein and flavocoxid (which is a mixture of various flavanols, primarily baicalin and catechin). Flavanols are very popular in certain circles of biomedical scamming, because of the money from wine and chocolate industry, and because the drugs are assumed to be safe because natural and from plants. Yes, there are limits to this logic.
Here one such flavocoxid paper by the Squadrito gang:
Antonio Micali , Giovanni Pallio , Natasha Irrera , Herbert Marini , Vincenzo Trichilo , Domenico Puzzolo , Antonina Pisani , Consuelo Malta , Giuseppe Santoro , Rosalba Laurà , Domenico Santoro , Francesco Squadrito, Domenica Altavilla, Antonino Germanà , Letteria Minutoli Flavocoxid, a Natural Antioxidant, Protects Mouse Kidney from Cadmium-Induced Toxicity Oxidative Medicine and Cellular Longevity (2018) doi: 10.1155/2018/9162946
Mice received heavy metal poisoning:
“The animals were divided into four groups to receive vehicle (0.9% NaCl) alone, flavocoxid alone (20 mg/kg/day i.p.), cadmium chloride (CdCl2, 2 mg/kg/day i.p.) alone , or CdCl2 (2 mg/kg/day i.p.) plus flavocoxid (20 mg/kg/day i.p.), respectively. […] All mice were sacrificed after 14 days of treatment with an i.p. overdose of ketamine and xylazine, and bilateral nephrectomies were performed.”
To obtain this fake microscopy image of a kidney:
But then Cheshire spotted something else:
“I am interested if the authors might address the issue of this or other patents on Flavocoxid, studied in this paper, which is available here. It appears the authors could have a relationship that should be disclosed to readers and peer reviewers.“
Suqadrito remained silent, but he did reply in another case of omitted COI, in Irerra et al Nutrients 2017 where Cheshire commented, referencing Squadrito’s CV:
“This paper’s conflicts of interest disclosure reads, “The authors declare no conﬂict of interest.”
Could the authors please describe the relationship between Dr. Francesco Squadrito and Primus Pharmaceuticals of Scottsdale, AZ, USA? In his CV, it states that he has been a principal investigator for this company since 2005, studying pharmacological potential of genistein aglicone. Is this a paid position? Is that position related to this paper? Does Dr. Squadrito receive royalties on the related patents?“
Squadriot replied on PubPeer:
I disclosed myself to Squadrito, but he didn’t explain me.
It is not just Squadrito who works as principal investigator for Primus. Altavilla does this since 2015, as per her CV. Bitto acts as “consulting researcher” for Primus since 2006 as per her CV, and she even founded her own company to market genistein, SunNutraPharma s.r.l.
If you believe Squadrito that he and his gang don’t receive money into their private pockets from Primus, you might just as well believe that their western blots are not fake.
Regarding this curative substance genistein from plant extracts, here another such COI-free paper:
Sonia Messina, Alessandra Bitto, M’hammed Aguennouz, Gian Luca Vita , Francesca Polito , Natasha Irrera, Domenica Altavilla, Herbert Marini, Alba Migliorato, Francesco Squadrito, Giuseppe Vita The soy isoflavone genistein blunts nuclear factor kappa-B, MAPKs and TNF-α activation and ameliorates muscle function and morphology in mdx mice Neuromuscular disorders : NMD (2011) doi: 10.1016/j.nmd.2011.04.014
“I am hopeful that the authors in the meantime could help readers understand the relationship between 1) the inventors of the patents, 2) the owners of the patents, 3) the authors, and 4) Primus Pharmaceuticals, who may own or have licensed the patents.
It seems likely that Primus has licensed the patents, but not who from. Nor is it clear whether the authors were compensated for the patents or the research related to the inventions (colloquially, but perhaps not legally, “drugs”) being studied, or whether these were owned by the University. It does appear that Primus was the sponsor of multiple clinical trials for Genistein and Flavocoxid conducted by the authors and that one or more of the authors was the principal investigator for Primus for at least one of these drugs. […]
While Primus is mentioned in many of the authors’ papers about these drugs, this particular paper seems to only mention the company in passing, “Genistein aglycone was a kind gift of Primus Pharmaceuticals Inc., Scottsdale, AZ, USA.” There doesn’t appear to be any other disclosures about whatever relationship might exist.“
Well, actually we do know from whom Primus licensed the patents. All these flavanol patents with Squadrito as inventor have the company Primus Pharmaceuticals as assignee. The only non-Messina inventors are Primus executives James Weir (President & CEO) and Bruce Burnett (left company). Here is a flavocoxid paper by the Squadrito gang with Burnett as coauthor (“approved by the Ethics Committee of the University of Messina“):
D Altavilla, L Minutoli , F Polito, N Irrera, S Arena , C Magno , M Rinaldi , B P Burnett , F Squadrito, A Bitto Effects of flavocoxid, a dual inhibitor of COX and 5-lipoxygenase enzymes, on benign prostatic hyperplasia British Journal of Pharmacology (2012) doi: 10.1111/j.1476-5381.2012.01969.x
Here, the rats “were treated daily with testosterone propionate (3mg·kg−1 s.c.) or its vehicle for 14 days to induce” prostatic hyperplasia and then injected with flavanol and other drugs. The experimental outcome:
Aneurus Inconstans noted:
In this study, transgenic mice were injected with flavocoxid, the results forged as usual:
Sonia Messina, Alessandra Bitto, M’hammed Aguennouz, Anna Mazzeo, Alba Migliorato, Francesca Polito, Natasha Irrera, Domenica Altavilla, Gian Luca Vita, Massimo Russo, Antonino Naro , Maria Grazia De Pasquale, Emanuele Rizzuto, Antonio Musarò, Francesco Squadrito, Giuseppe Vita Flavocoxid counteracts muscle necrosis and improves functional properties in mdx mice: a comparison study with methylprednisolone Experimental Neurology (2009) doi: 10.1016/j.expneurol.2009.09.015
The mice in this flavocoxid study were less lucky, they were made to suffer and die from intestinal sepsis:
Alessandra Bitto, Letteria Minutoli, Antonio David, Natasha Irrera, Mariagrazia Rinaldi , Francesco S Venuti , Francesco Squadrito, Domenica Altavilla Flavocoxid, a dual inhibitor of COX-2 and 5-LOX of natural origin, attenuates the inflammatory response and protects mice from sepsis Critical care (2012) doi: 10.1186/1364-8535-16-r32
“Specifically, mice were anesthetized with ether, and a midline incision was made below the diaphragm to expose the cecum. The cecum was ligated at the colon juncture with a 4-0 silk ligature suture without interrupting intestinal continuity. The cecum was punctured once with a 22-gauge needle. The cecum was returned to the abdomen, and the incision was closed in layers with a 4-0 silk ligature suture. After the procedure, the animals were fluid-resuscitated with sterile saline…”
To produce this fake preclinical data in support of flavocoxid:
Despite the aforementioned patent for flavocoxid and other flavonols, the Squadrito gang wrote:
Let’s have more COI-free flavocoxid!
Here, rats were tortured to prove some curative powers of flavocoxid: kainic acid (KA) was injected into their brains to cause symptoms of epilepsy.
Letteria Minutoli, Herbert Marini, Mariagrazia Rinaldi , Alessandra Bitto, Natasha Irrera , Gabriele Pizzino , Giovanni Pallio , Margherita Calò , Elena Bianca Adamo , Vincenzo Trichilo , Monica Interdonato , Federica Galfo , Francesco Squadrito, Domenica Altavilla A dual inhibitor of cyclooxygenase and 5-lipoxygenase protects against kainic acid-induced brain injury NeuroMolecular Medicine (2015) doi: 10.1007/s12017-015-8351-0
“After general anesthesia with sodium pentobarbital (50 mg/kg i.p.), the animals (n = 96) were divided into the following groups: sham + vehicle (1 ml/kg i.p.); sham + flavocoxid (20 mg/kg i.p.); KA (10 mg/kg i.p.) + vehicle; and KA + flavocoxid (20 mg/kg i.p.). Flavocoxid was administered 30 min after KA injection.”
The result was some fake western blots:
So you see, Squadrito did a great job helping Primus to preclinically verify their drug flavoxid, which the company markets as “medical food” under the tradename Limbrel®.
And then, this happened, an FDA announcement from 2018:
Primus Announces a Voluntary Nationwide Recall of All Lots Within Expiry of Prescription Medical Food Limbrel® Due to Rare But Serious and Reversible Adverse Events While Seeking FDA’s Cooperation to Restore Access for Patients with Medical Necessity
This recall has been completed and FDA has terminated this recall.
“Primus Pharmaceuticals, Inc. of Scottsdale, Arizona is voluntarily recalling all unexpired lots of Limbrel products to the patient (user/consumer) level at FDA’s request. FDA has requested a recall of Limbrel due to rare but serious and reversible side effects associated with Limbrel.
Between January 1, 2007, and November 9, 2017, FDA received 30 adverse event reports of elevated liver function tests or acute hypersensitivity pneumonitis associated with the use of Limbrel products. These conditions present in rare cases with varying degrees of severity in patients taking Limbrel for the first time in the initial weeks of exposure, and may go unnoticed by the patient until they consult with their physician or until symptoms develop that require hospitalization. There have been no deaths reported with the use of Limbrel, and in all reported cases adverse effects resolved without residual effects after discontinuing use of the product. [….]
Limbrel has been marketed since 2004 as a medical food available only by prescription for patients under active and ongoing supervision of a physician for the dietary management of osteoarthritis (OA), a degenerative disease of the joints and the most common form of arthritis. Prior to marketing, Primus conducted clinical studies that support the efficacy and safety of Limbrel and compiled an extensive dossier providing an analysis of published data to support the medical food status of Limbrel and to establish how the product meets the distinctive nutritional requirements of OA. Primus stands by the legal status of Limbrel as a medical food. Limbrel products have been distributed nationwide in the USA to wholesalers, pharmacies, and physicians as medical foods without challenge from FDA for over 13 years, with approximately 2 million prescriptions and physician samples dispensed to an estimated 450,000 patients.”
There are now several law firms inviting those damaged by Limbrel to a class action lawsuit against Primus. Sadly, Squadrito’s rodents were tortured to death and can’t sue.
It wasn’t of course just mice, rats and gerbils which Squadrito and his gang deployed to help Primus market their jointly patented drug. There also was a clinical trial at the University of Messina, to treat Duchenne Muscular Dystrophy with the flavanol quackery.
Gian Luca Vita, Maria Sframeli, Norma Licata, Alessandra Bitto, Sara Romeo, Francesca Frisone, Annamaria Ciranni, Giovanni Pallio, Federica Mannino, M’Hammed Aguennouz , Carmelo Rodolico, Francesco Squadrito, Antonio Toscano , Sonia Messina, Giuseppe Vita A Phase 1/2 Study of Flavocoxid, an Oral NF-κB Inhibitor, in Duchenne Muscular Dystrophy Brain Sciences (2021) doi: 10.3390/brainsci11010115
Of course the authors declared to have no conflicts of interests while being listed as patent inventors. Cheshire noted, among other things:
“The registered clinical trial NCT01335295 does help with the timeline a bit. Looks like trial commenced April 14, 2011 and ended December 2013.
The trial page shows 20 people were enrolled. This paper discusses 17 people were treated with flavocoxid and 17 were not. Could the authors clarify the study cohort sizes?“
The trial NCT01335295 listed our friend Giuseppe Vita as principal investigator (his son Gian Luca is first author). Vita Senior explained on PubPeer:
“Ethical approval was obtained early in 2011. At that time, no specific reports of liver injury due to flavocoxid were available except rare elevations of liver enzymes in some trials. The protocol included serum AST, ALT and LDH determinations.The patients were enrolled and followed in the period 2011-2013. […] The owner of the patent is Primus Pharm. […]
When the trial was registered as NCT01335295, the protocol designed the recruitment of 20 patients, all to be treated. Since it was an open-label study, and also after presentation to TREAT-NMD Advisory Committee for Therapeutics (TACT), we then decided to include an untreated group. The number of 20 treated patients was not reached and we stopped at 17 treated subjects and untreated groups same size.“
The trial’s participants were children suffering from Duchenne Muscular Dystrophy, aged “between 4 -16 years”. Having fabricated a massive amount of papers with fake preclincial data about flavocoxid, this Messina gang thought it was a good time to switch from rodents to children. Who then received a liver-damaging drug.
I am sure the ethics committee of University of Messina finds all this hilarious. As reminder, the university never replied to my emails. They don’t give a flying f***. Patient abuse in bullshit clinical trials? Meh.
Finally a proper reaction from University of Messina. Squadrito now announced that he is suing me in court. Because he obviously disagrees with FDA’s decision that Flavocoxid/Limbrel is toxic. Here his email:
“You sent the below mail on May 2nd 2022 to more than 50 coauthors stating that I also experimented on children with a toxic substance [actually, the email was addressed to 50 recepients including trial’s PIs- LS]. This is a false and serious allegation. It is not a problem of blots. This accusation has penal l implications. You cannot send statements of this kind to everyone. With this crazy allegation of yours you are accusing me unfairly and you have created serious moral and material damage for me that you must compensate. You have to pay for it. My attorney is suing you and will demand appropriate money compensation for all the damage you have done to me with this false allegation without proof. People you emailed will testify against you. I am looking forward to meet you in the court.
Francesco Squadrito “
Money, money, money, that’s all he can think about, this “scientist” without any COI. Go sue FDA then.
As Squadrito kept announcing lawsuits in his follow-up emails, he said this interesting thing:
Domenica Altavilla, wife of Squadrito, has suddenly died.
University of Messina announced the funeral for 14 May 2022. It is a bizarre obituary. It mentions her h-index, 300 papers, her work for Primus since 2015, a company she founded (“SunNutraPharma S.r.l.”, unmentioned other owners Squadrito, Bitto, Primus Pharmaceuticals, Pizzino & Irrera), other pointless things like curcumin, but the obituary never states any cause of death, or how old Altavilla was when she died (which was 64). Neither is Squadrito mentioned.
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Another possible concern with some different authors as University of Messina. https://pubpeer.com/publications/A3542BF3E1DAC7A42C50C4A87E8AEA
Retraction, 10 October 2022:
“Retraction to: Letteria Minutoli, Antonio Micali, Antonina Pisani, Domenico Puzzolo, Alessandra Bitto, Mariagrazia Rinaldi, Gabriele Pizzino, Natasha Irrera, Federica Galfo, Salvatore Arena, Giovanni Pallio, Anna Mecchio, Antonino Germanà, Daniele Bruschetta, Rosaria Laurà, Carlo Magno, Herbert Marini, Francesco Squadrito, and Domenica Altavilla “Research Article Flavocoxid Protects Against Cadmium-Induced Disruption of the Blood-Testis Barrier and Improves Testicular Damage and Germ Cell Impairment in Mice” Toxicological Sciences, Volume 148, Issue 1, November 2015, Pages 311–329, https://doi.org/10.1093/toxsci/kfv185
In May 2022, a reader raised concerns about Western blot bands in Figures 1, 2, 5, and 7 of the article. Details of the concerns were published on PubPeer: https://pubpeer.com/publications/A145E6D3AEFFAA20065285BAFCDCAD.
Subsequent examination of the figures showed clear evidence of duplication for parts of the images. The authors are unable to provide the original data for inspection and have not provided satisfactory explanations to the concerns raised. The extent of the manipulation and the impact on the study conclusions are considered extensive. Therefore, the article is retracted in accordance with image manipulation guidelines endorsed by the Committee for Publication Ethics (COPE).”
Wait, there’s more!
Retraction for Bagnato et al. 2013 by Arthritis Research and Therapy, today 4 December 2022:
If I`m not mistaken, the Fraud Squad currently has 3 retractions and 3 expression of concern. And counting.
Bravo ‘Actinopolyspora biskrensis’!
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Posted concerns about 2 more papers today by the Fraud Squad: https://pubpeer.com/publications/FA3889E8BF5C99A6DF7E95BB6F6378 and https://pubpeer.com/publications/5C59D58152694098EB92CB39C4F196 (seems to be a duplicated image across papers, described differently).
Summary as of today 21 March, 2023, for The Fraud Squad.
Seven Expression of concern:
Needless to say we wish many more to come.
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Three more Retractions, all from the journal Critical Care medicine:
Thus, 8 retractions and 7 EoC is the new total for the Fraud Squad as of today 25 March, 2023.
Another retraction, this time from the journal Critical Care, to Bitto et al. 2012:
9 retractions and 7 EoC for the Fraud Squad as of today, 19 Apr 2023.
Tenth RETRACTION for the dreadful Fraud Squad, 2 June 2023, to Altavilla et al. 2012 Br J Pharmacol:
The above article, published online on 3 April 2012 in Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement between the journal’s Editor-in-Chief, Péter Ferdinandy, the British Pharmacological Society, and John Wiley & Sons Ltd. The retraction has been agreed due to evidence of image duplication and manipulation. Specifically, the western blot in Figure 4B has been previously published in another article (Bitto et al., 2010) purporting to display a different protein. Furthermore, two signals are identical of which one is 180 degrees rotated, and b-actin signals have been duplicated. Finally, the b-actin blot of Figure 4C has been previously published in another article (Bitto et al., 2010). As a result, the conclusions reported in the article are not considered reliable.