This story spans several years between 2010 and 2014. It was sitting in my drafts folder for over a decade, so some links may be dead and the current status of various people or companies or science may no longer be accurate.

Here’s the short version…

• My lab got a drug from a company via a material transfer agreement (MTA)
• We found some really cool results
• We tried to publish results
• The company tried to sue us
• The company went bankrupt
• We published the paper anyway
• Rival paper from former company employees came out, doesn’t cite us

Background on Conflict of Interest in Basic Science
For life scientists in academia, one of the most common examples of potential conflict-of-interest (COI) is receiving resources from a drug company to do a research project. Sometimes this takes the form of money directly intended to fund the research. Other times the reward can be a position on the scientific advisory board of the company, or being granted access to rare chemicals/reagents that are not commercially available to everyone. In the US, such support is usually required to be disclosed both to the employing University, and federal or other funding agencies.

Usually when the resource being shared is a new drug, companies like to control what happens to the material, such as whether/how the academics can discuss the results with others, including publication. Such legal matters are usually dealt with via Confidentiality/Non-Disclosure Agreements (CNDAs) and Material Transfer Agreements (MTAs). These documents are drafted up jointly between the lawyers in the company and the University’s Technology Transfer Office. Herein, I discuss an example of such an agreement gone awry….

The research that led us to request an MTA for a new drug
My lab’ has a long-standing interest in mitochondrial K⁺ channels, and in our work we’ve often used a drug called NS1619, which is reported to specifically activate mitochondrial K_Ca channels. The nomenclature is a bit odd here… a family of channels is encoded by the Slo genes, of which there are 4 isotypes in mammals (Slo1, Slo2.1, Slo2.2, Slo3). The channel proteins are also called “BK” channels, since they are large conductance (big) K⁺ channels. They’re also often referred to as K_Ca channels, even though the Slo2 variants are not Ca²⁺ activated (they’re Na⁺ activated, so K_Na channels). There are also some hybrid terms such as BK_Ca, or mBK (mitochondrial BK). Anyway, the important thing is these channels are thought to be important for protecting the heart against ischemia-reperfusion (IR) injury, and the debate mainly centers on which isotype is present in mitochondria and responsible for these effects. Other people think it’s Slo1 but our results suggest otherwise.

NS1619 was originally made by a Danish company called Neurosearch (archived link from 2019 shortly before site went dead), but is readily available from chemical suppliers such as Sigma. However, over decade or so since the drug was made available, it emerged that NS1619 might have non-specific side effects in mitochondria. So, interest in the field shifted toward a related newer compound, NS11021, reported to be more potent and specific for the mito’ channel. NS11021 is not commercially available, so in late 2011 we wrote to Neurosearch to obtain some. Following a few rounds of back-and-forth between lawyers, an MTA was executed and shortly afterward we were sent an aliquot of NS11021. The scientist within the company who liaised with us, Morten Grunnet, was collaborative and very open to discussing experimental details and results. I consider him a colleague and friend to this day, and he was a co-author on our paper that came out of this work.

Although publishing the entire MTA here would be inappropriate, below I want to highlight specifically the part describing the work to be performed in my lab using the drug. Note the mention of potential use of knockout mice for various types of K⁺ channel…

Appendix A: Study Protocol. The overall direction of the research project, which the study will be part of, is the identification of K+ channels in mitochondria, which are involved in cardioprotection by ischemic or anesthetic preconditioning. The aim of the study would be to investigate the role of the BK channel in anesthetic preconditioning using FVB mice. In Langendorff perfused rat hearts, the volatile anesthetic isoflurane provides protection against ischemia-reperfusion injury in a manner that is sensitive to the BK channel inhibitor paxilline.  University of Rochester would like to determine if this manner of isoflurane mediated protection can be mimicked by the administration of the specific BK channel activator NS11021. University of Rochester would also like to look at the effect of NS11021 on BK channel activity in isolated mitochondria, using novel thallium-flux assay we recently published (PMID:20185796, PDF attached). None of the K+ channels in the mitochondrial membrane have been identified at the molecular genetic level. Knockout mice for various types of KCa and KATP channels may be available to us in future, and knowing which channel to look for could be greatly facilitated by the use of highly specific pharmacologic reagents.

So, we did some experiments and the data looked great. Without getting into details (all of which can be found in our published paper on this topic), the key finding was that NS11021 protected hearts against IR injury, and this effect was lost in Slo1^-/- mice. Furthermore, the effect was likely mediated not by Slo1 channels inside mitochondria of cardiomyocytes, but instead by channels in intrinsic cardiac neurons.

Things get nasty
You might think if a company was touting a drug as acting on a specific target, and someone came along with data showing the effect of the drug disappears in a knockout mouse for the target, they’d be happy?  Not Neurosearch, they lawyered-up and tried to stop us from publishing.  They essentially said that if we attempted to publish, it would be a breach of the MTA, and they would sue for breach of contract.

Backing up for a second, during the time we were collecting our data and preparing our manuscript, we were in regular discussion with Dr. Grunnet, who was essentially our contact within the company. However, in the summer of 2012 he left Neurosearch to work for a different Danish Biotech, Lundbeck. We continued communicating while the manuscript went through several iterations, then in June 2012 we heard via Dr. Grunnet that Neurosearch were “rather reluctant to let us publish since they claim that the part including KO mice is not covered by the MTA”.  Also “they have decided to perform a small study on 11021 in WT and BK KO mice with I/R in Langendorff. The primary aim is to confirm BK selectivity of 11021 and they are aiming at a small publication”. The suggested solution was that we hold off until their study was published. In other words, the company scientists didn’t want to get scooped!

Given our study was complete and ready to publish, and theirs had barely even begun, we tried explaining that it might be better to simply add the interested parties onto our paper as co-authors. In my view this was an outreach effort that went above and beyond necessary collegiality (remember – they were trying to sue us). However, we were then contacted by Søren-Peter Olesen who oversaw Neurosearch’s collaboration with the University of Copenhagen, and also holds an academic appointment there.

In the remainder of fall 2012, while our paper was being put through the wringer by journal reviewers, a conversation ensued between myself, Dr. Olesen, and the Tech’ Transfer lawyers here in Rochester. This culminated in an email from Dr. Olesen in November, stating “Neurosearch will then close the matter and conclude that you do not want any legal permission to publish on NS11021 in relation to transgenic animals”.

We had the University lawyers look over the MTA again, and they made two important conclusions… First, the language was sufficiently broad regarding knockout mice that in their view we hadn’t breached the terms. Second (and more important) the company had no right to bar us from publishing because the MTA itself made no mention of a right-of-veto on publications. The MTA simply requested we send a copy of any manuscript to Neurosearch 30 days before journal submission, for them to review and comment. We did this, and in-fact our company point-person had been kept in the loop from the earliest stages, so Neurosearch knew about these knockout mouse studies for almost a year before the paper was submitted.

Neurosearch goes kaput
Eventually, after 4 journals and 8 months of review/reject/resubmit, we got our paper published in Peer J  in February 2013. But then something strange happened… Neurosearch went belly up. Well, technically the terms used were restructuring, transfer of assets, prosecution for share price manipulation. So, the company that threatened to block us from publishing no longer exists.  Another company trading under the same name and with new management did emerge from the ashes of the old Neurosearch, but disappeared in 2019.

What about the company-backed study?
Olesen and colleagues finally published their version of the story in PLoS One, in collaboration with a group from the University of Tübingen. Although the new paper cites our work, there are a number of problems with it…

• The paper purports to show patch-clamp data of BK channel activity in mitoplasts (isolated mitochondrial inner membranes). The problem is, they used a “Port-a-Patch” system from NanION. This system works by using a vacuum to pull down a spherical object (ideally a mitoplast) onto a pre-formed patch pipet, with no microscope or any other confirmation that what you’re actually patching IS a mitoplast. The method depends on the purity of the “soup” you put into the chamber. We tested one of these systems in my lab in 2012 and determined it was useless for mitoplasts. Any contamination with other membrane fragments gave false readings, and notably the PLoS paper contains no information on the purity of preparations used for patching. The image of mitoplasts in Figure S1 shows a lot of membrane fragments apart from mitoplasts.
• The IR injury data concludes that hearts from Slo1 KO mice cannot be protected by ischemic preconditioning. This experiment is completely opposite to our published findings. No attempt is made to explain this (remember, they don’t cite us) but here are some suggestions… we did everything in male mice, on a single genetic background (FVB), whereas they used both male and female mice in a mixed background (Sv129/C57BL/6). We measured cardiac function with a pressure balloon, but they only measured heart rate, so did not have any functional data for the heart perfusions. They had a 4 minute delay on ice between heart extraction and perfusion. The mouse heart is exquisitely temperature sensitive, and our delay is typically <30s. with no ice. Any longer and contractility is compromised.  In effect, they may have been looking at hearts with drastically compromised function, before any IR injury.
• We use a constant flow system so the heart is always sufficiently oxygenated and function is not affected by coronary vascular tone. The Lukowski study used constant pressure in which coronary flow (O2 delivery) is affected by vascular tone. They showed that IPC improved post-IR coronary flow, and this was absent in BK knockouts. It cannot be ruled out that the knockout mice had compromised post-IR coronary flow. This links the effects of BK in IPC to coronary vasculature, NOT mitochondrial channels inside cardiomyocytes.
• The dose of NS11021 used in the patch-clamp studies is very high (10 μM). Previously we showed 50 nM could activate a paxilline sensitive K⁺ channel in mitochondria. The PLoS paper claims that their patch data complement findings of another paper touting Slo1 as a mito’ BK channel, but that paper didn’t show any patch data.

Overall not a good situation. Sufficient time has passed since these events that it’s probably OK to talk about it now. Olesen is retired/emeritus. The company that NS sold some of its IP/assets to is still around, but has yet to bring a single product to market based on the Neurosearch drugs.  The lawyers involved have all moved on in their careers. My lab no longer does much work on mitochondrial ion channels (but we do have some unpublished data).  People are still publishing on NS11021, and would I doubt that any of the more recent folks using this molecule are aware of the above troubles.

Continuing the saga of Alan Cash and Terra Biological, trying to get a dietary supplement containing oxaloacetate into clinical trials for all sorts of conditions from long-covid to PMS, there appears to have been a “breakthrough“!

The company finally got around to publishing the results of a clinical trial on the effects of OAA on self-reported symptoms of chronic fatigue syndrome / myalgic encephalomyoelitis (CFS/ME).  Now, this is a condition that scientist George Monbiot has called “The greatest Medical Scandal of the 21st Century”, so straight away that should start raising flags. Why pick a poorly-defined condition, the very existence of which is hotly debated?

Prima facie, the results appear promising and statistically significant, and it’s also commendable that the group chose to provide the complete (not actually) data set.  However, digging into the data there are are several problems…

The number of patients is “wobbly”

The trial started out with 40 patients in the control arm and 42 in the treatment arm. However, there was apparently a greater attrition rate in the controls (12 leaving) than the OAAs (5 leaving).  This would leave 28 controls and 37 OAAs as “completers” of the trial, as nicely explained in the flow diagram in Figure 3 of the paper and in the manuscript text…

…which makes it a bit weird when we go to the original data set (re-hosted here) and see there are actually 29 controls!  Where did the extra control patient come from?

Biased reporting of patients who got worse

Things get real squirrelly when we look at Figure 5 of the paper. The y-axis here is number of patients, the x-axis is the change in fatigue score pre/post trial. The orange are controls and the blue OAAs.

It’s a fairly simple matter to look at the dots and count the number of patients at each score point, then add them up. Doing so, we again arrive at 29 for the controls. BUT, importantly there are only 35 patients in the OAA set. We’re missing two of them, and by looking at the original data we find there were indeed two patients on OAA whose scores got worse (-4). They were simply eliminated from this graph, making it appear as if no patients on OAA got worse.

There are other discrepancies between the data and this graph, as shown in the table below. Anything highlighted blue doesn’t match up. There are 3 instances where a ‘1’ was assigned to the control group in the figure, suggesting a patient got worse, when in-fact there was no patient getting worse at that score level.  Furthermore, there are 2 patients who got better in the control group but were not counted on the graph…

Plotting Figure 5 as shown in the paper (left image below) alongside the real data (on the right) shows the way in which the paper makes it appear the controls got worse. In the published version there are more orange (control) points above the x-axis on the left side of the graph (worse scores).  Notice the missing 2 OAA patients who also got worse (at -4 in the right hand graph).

Just for fun, I also switched the order of the data series, so now the controls (orange) appear on top of the OAAs (blue) in the “real” graph on the right. This highlights the 3 controls who had a big improvement of 11 points… the same as the OAA group.  It’s amazing what little differences like this can make to the perception of a result.

Trial non-completers?

Now remember, Figure 5 is only the folks who completed the trial. As already mentioned, many did not. By comparing the data from the completers vs. the whole set, we can gain some more insight into the non-completers group, as shown in this table below.

What this shows, is of the 11 controls who quit early, they had an average improvement score of 2, and none of them got worse during the trial. However, of the 5 in the OAA arm who quit early, 3 of them actually got worse.

Combining the analysis of the completers and non-completers, overall in the control group 7 patients got worse during the trial, but none of them quit early. However, in the OAA group 8 patients got worse during the trial, and 3 of them quit early. Readers can judge for themselves whether there may have been any sort of “encouragement” applied to certain groups who were feeling worse to quit the trial early, but no such encouragement applied to those in the other group.

Appropriate Statistical Tests

Lastly, I’m not a statistician but my understanding is that when testing if 2 groups are different from each other, without knowing in advance which direction any difference might be, you should use a 2-tailed T-test.  For some reason, here the authors chose to use 1-tailed tests for everything, i.e., they only hypothesized that the results would go in one direction (presumably OAA being better than control), and not the opposite possibility. Needless to say, if you re-do the tests applying the proper criteria, some of the differences get a lot smaller or vanish altogether.  For example, in Figure 4 the p-value of 0.057 is described in the text as “trending toward significance”.

Performing this test with 2 tails yields a far-less impressive p-value of 0.114, quite literally nothing to write home about.

Wrapping this up

Let’s not get into the numerous typos in the paper that really speak more about the shitty editorial standards at Frontiers than anything else. Further illustrating such problems – one person who reviewed the paper (Alison Bested) has published with at least two authors on the paper (Yellman and Bateman) as recently as 2021, so it was reviewed by familiar folks. The discussion of the paper repeats many of the mistakes regarding the simple biochemistry of OAA that I’ve written about in the past. Bottom line, yet again, the company shilling a $600 a year supplement has managed to get something published with a veneer of scientific legitimacy in a not very good (predatory?) journal. A not-very-deep-dive shows problems with data reporting and the basic arithmetic keeping track of numbers of patients. Don’t human patients with hard-to-diagnose-and-measure chronic diseases deserve better than this?

It’s been a while! Several updates to the lab over the past few months…

• Rahiim Lake passed his PhD qualifier exam last fall (congrats!)
• We hosted the 11th annual TRiMAD (Translational Research in Mitochondria Aging & Disease) conference here in October, attended by over 160 people.
• Paul spoke at a number of conferences and outside institutions, including a conference at UPenn on research integrity, resulting in this paper.
• The University finally adopted and published the Open Access Publishing Guidelines that we worked to bring through the faculty senate. It only took 7 years!
• Paul went back to school in the Executive MBA program at UofR’s Simon School of Business (anticipated graduation May 2026). It resulted in finally (reluctantly) getting a linked-in page.
• Post-doc Sabarna Chowdhury got married in April 2025 (congrats!)
• Lots of other papers got published, including some exciting work on taurine metabolism with the Bajaj lab in the (newly NCI designated) Cancer Center, which is accepted for publication in Nature.

A rotary evaporator (RotaVap) is a core piece of equipment for any lab that does chemical synthesis, and Buchi make some of the best ones. Unfortunately, ours is about 25 years old and they no longer make parts for it.

In the heart of the apparatus there’s a worm-drive… the motor is connected to a helical spindle which drives a ring gear. This is what spins the round-bottomed flask.

Over the past few months we’d been noticing that at low spin speeds, the rotation would skip.  It would intermittently speed up and slow down through the rotation cycle.  So, we opened up the gearbox to see what was inside (note – you may need to use a hammer to get the parts to separate out).

The problem is, Buchi decided to make the gear out of plastic, and years of exposure to grease and chemicals causes the plastic to crack, as shown here…

The result is the motor spins but the plastic rung just grinds around and doesn’t spin the metal part that’s eventually attached to the flask.  The worm gear is basically slipping once per rotation. We tried buying a used model off eBay, and upon dissection it had exactly the same problem, so it appears this is a common fault on this particular model (RE-111).

The solution is not as simple as “glue it back together”. There’s too much grease everywhere, so it needs completely stripping down to remove all that so the glue will stick. And then, removing the ring gear can only be done by snapping it in two. The problem then is there’s not much surface area to glue back together so it will probably break again soon.

Enter the 3D printer!

With some quick work in Sketchup  I was able to make a model of the gear with a break in the middle and plenty of surface area for gluing the 2 halves together, as well as gluing to the metal spindle inside the machine.

Here’s the new gear, printed in Elegoo ABS-like resin on a Mars 3 printer. The STL file is here to download if your Buchi Rotavap is in need of a fix!