After only giving the study a brief look at the time, and in light of the missed and somewhat confirmatory c60oo life-extending findings of BuckyLabs, I decided to take a more detailed look at Ichor's c60 paper.
The study design appears to merit considerable criticism, as well in the reporting of its findings. It seems as though the data was filtered to produce an eye-catching press release, rather than one which would accurately report unglamorous, conflicted, and so unresolved, findings.
Longecity studies, especially in clinical trials, where studies are not concluded often focus on the median, but the mean where can be calculated of course matters, especially if there are no extreme outliers skewing the results. The median contextualises the mean.
There was no presentation of mean data and apart from a lazy nothing-to-see-here reference and no supplementary data provided to enable the reader to do so. There was, though, a clear life-extending effect with c60oo in both experiments over oo (olive oil).
The area under the curve for the older mice injected with c60oo intraperitoneally (IP) is evidently larger - so more life and so longer average lifespan. Without the lifespan data, all that is left is to visually estimate: 6 weeks per c60oo mouse over their olive oil consuming counterparts was the guess.
When you're after 90%, what does it matter whether it's 2 or 8%: mice could collectively sneeze themselves into double digit life-extension. Well, proving that c60oo is life extending in mice matters, and it matters a great deal. The importance was perhaps lost within the community anticipating replication of Baati, unable to reframe or reimagine Baati. However, it shouldn't be lost on scientific rigour driven researchers for whom professional process and reporting of findings pays the bills.. But it was unacceptably and inexplicably overlooked.
So the first study looks a little like 6 weeks life extension for c60oo over oo which equates to something like 4% absolute of lifespan gain, and a study duration gain of maybe 15 - 20% (the gain from the beginning of the lifespan study).
In the second expanded study, this time with oral administration and dual intervention ages (though lifespan measurements start from the same age) there is clearly once again (unreported, unrecorded) life-extension of c60oo over oo. The visual guesstimate was of a lifespan gain of 5 weeks. A different strain of mice, these rodents were shorter-lived than their IP-counterparts. A little shy of 4% on total-lifespan with something similar on effective study extended lifespan of around 15+%).
While the following is not an effective reduction of the studies combined, due to in particular, the intervention split age start dates of the second study, consider the following 60 mouse study: 30 intervention, 30 control starting at 110 weeks for a species with expected lifespan of 140 weeks and one where the intervention group lived on average for 145 weeks. As cited there are naturally flaws with this simplification, but it is a reasonable comparison. Lazily, I ran this through both AI chats and they both reached the same conclusion: 97% confidence of the intervention being life-extending. So a 3% chance the intervention is not life extending.
The studies combined should run close to this - it could fall a little short of 95%.
Now is it reasonable to assume Ichor did not do this analysis, a simple mean life expectancy and confidence test? Given that does not seem likely, lest the authors be incompetent, did they not see fit to make the data available for those interested in the study to do the test, and to those within a community who partially funded it? They plainly dodged it.
We have this:
"Since olive oil and many other oils may form lipid granulomas when administered by IP [36], it may be hypothesized that the lack of efficacy observed in the lifespan study on CB6F1 mice may be affected by this phenomenon. However, the mean lifespan of treated and untreated mice were comparable to lifespans reported by others for CB6F1 mice (126–143 weeks) [37], and even with the oral supplementation, we have not been able to demonstrate a clear effect on lifespan. In support of the idea that lipid granulomas were not a significant cause of the acute lethality of these injections, previous works have demonstrated that injections of similar volumes of olive oil had relatively little ill effect""
So no mean lifespan data, just that it is similar to other lifespans of this strain of mice. And yes, there is a clear demonstrable effect of c60oo on lifespan within the study over olive oil, Ichor just failed to report it.
So there is clear evidence that "c60 in olive oil extends life in mice over olive-oil controls" rather than c60oo does not extend lifespan in mice, as the study claims. The only controls in the study were those treated with olive oil: yet the lifespan comparison was unreported.
This purported 4% is not simply a knot for the longecity community to cling on to, it is a critical and powerful finding which likely provides supporting evidence to Baati and Moussa’s extraordinary claims, as unlikely as it seems: it just didn't feel that way for those of us who had harboured imaginings of c60 becoming the Higson-Boson biology. The community just needed to reframe, and Ichor failed to disclose findings which might have helped enable this transition.
One response to this criticism would be: well so what they didn’t beat controls, and significant life-extension above controls is what we’re for. It would seem puerile to state there within study no controls, just external ones, so they couldn’t ever beat internal controls. This study was about 90% life-extension replication, the use of external controls over internal ones are not going to blur this effect. Utilising mice typically reserved for a control group for a larger interventional group or to resource another study would seem wise, and in retrospect was proven so.
Controls were never going to seem that meaningful, if we have Baati, we have Baati: if we don't, we don't. Controls were never going to critically prove or disprove Baati & Moussa.
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We didn't get Baati, though. Blinded by the deflation of C60’s “God molecule" status, the community, abetted by Ichor, largely overlooked a key finding from the study. They might have dwelled longer on this result had Ichor as they were duty-bound to do reported c60 in olive oil significantly extended lifespan over olive oil in mice.
Once c60oo isn't Baati life-extending or even life-extending over typical strain lifespan, all that subsequently matters is the relationship between c60oo and olive oil (oo) on lifespan - and it matters a great deal. Within this context, the worst outcome for attempts to seek partial validation of Baati or to begin constructing a new function of c60oo on life-extension incorporating both studies would be to find no difference between the c60oo and oo groups respectively. That in effect c60 does nothing. This is precisely the message signalled when Ichor stated median lifespans to be almost identical: nothing doing here (there clearly was).
Better or worse is better than nothing for Baati-validation since C60 does something on lifespan and as such the study attests to something to solve, untangle, for C60oo on lifespan. Well, from lifespan curves, we had c60oo as significantly better than oo, yet it was unreported, just indifferent medians. C60 in olive oil was significantly decoupled from olive oil for impact on lifespan - yet not one line in the study write-up reports this extremely important partially Baati-validating fact.
Suppose Ichor's work had preceded Baati and Moussa, with the life-extending effect of c60oo over oo reported. How would the longevity community view Baati within the framing of Ichor's results, dismissively? No, context matters - the study-order matters, it would seem. Amidst the speculation there would be one word repeatedly surfacing on the community's lips: hormesis. Ichor would have been seen to have revealed a piece of the c60oo-lifespan relationship which the remarkable Baati study expanded - not considered to be a result in retrospect to be some advanced, a priori debunking of Baati.
Were c60oo proven to be hormetic it would have been of little surprise, and was a cautionary voice in the community against high dosing. Much of that which benefits health is hormetic, so why not c60oo? And given the extraordinary claims of Baati and Moussa, it should not be mystifying were the conditions enabling this dramatic life-extension revealed to be finely calibrated - that there was a sweet spot for lifespan extension in Wistar rats, which resembled the hormesis curve.
Returning to Baati - and that result. If proven true, one of the most remarkable findings in lab research was announced by Moussa and Baati in 2012 through accident, not design. The c60oo rats live nearly twice as long as controls, with no sign of cancer, typically prevalent in this breed, it is asserted, and purple livers.
What would be a professional scientific response to these findings in a world driven towards altruism? These guys were and are not liver, cancer or longevity experts - in publishing these results they perform "a handover" to those scientific communities of interest duty-bound to investigate these claims. It would be expected within this idealistic worldview, for representatives of these fields to pour over the work, seeking every study detail, however trivial it may seem, like a good detective taking down an eye-witness statement. Many scientific breakthroughs are accidentally discovered, this could be one of them. There is one key objective: verification. And so replication. Once established, then explore. Nothing apart from resource limitation, would prevent exploration and replication to be parallel experimental bedfellows. Exploration, as with Ichor, through a range of experimental alterations, could, given Baati's extraordinary results, have held the potential to verify despite the differences (i.e. with doubling the lifespan of mice). Ichor, though, should never have been considered as a verification-test. As such iIt could never disprove Baati, only prove it.
Ichor should not so much raise doubts over Baati's findings, but more so the conditions upon which those results were generated. And again it should have been stated that c60oo clearly demonstrated a life-extending effect over olive-oil controls in this study but wasn't not stated.
Ichor did not replicate. And though making cursory and obligatory references to study distinctions, Ichor through unqualified and undiluted headlines quick to communicate considerable doubt over C60oo life extending potential. They could have chosen a different headline: c60 in olive oil extends lifespan over olive oil controls in mice. And it did. Instead, Ichor went for C60 does not extend lifespan in mice: no nuance.
When Baati/Moussa announced their findings the (effective) response of the then guru of the longevity community when asked to comment: they screwed up the slides (see correction), so we have to doubt the findings of this study i.e. did the rats really live this long? This looks like resentment towards others achieving through accident, what they wished to do by design. Putting it politely it appeared a scientifically incurious response, but it was inexcusable, appearing cynically motivated. The researchers, and the paper's reviewers, were lazy, perhaps momentarily incompetent - so it should be assumed these scientists with decades of reliable experience, were not reliable reporters to the doubling of rat-lifespans: an absurd deduction To ignore this study, and its remarkable claims on lifespan and health, would require a 99.9% confidence it to be false - there was no such basis. The only people interested in replicating this study were the community of longecity - it was a citizen science project.
Ichor was no replication. The rats were supplemented with c60 mid-life, well for most Wistar rats mid-life, for 7 months: between months 10-17 (Reference, Ref: Moussa interview. These mice, who were not knowingly rats, were given c60 until the end of their lives - many starting out late in life.
The study is clearly meaningful in design, but a clear failure of replication. And when a study which looks a lot like an attempt to replicate fails to replicate, the failure of replication id broadcast - no matter the obligatory and almost trivial disclaimers (Baati was all male, we were mixed sex) with a message of "we were never trying to replicate, but we replicated enough to bring into serious doubt Baati's results".
Ichor didn't hold back, there was no tempered study headline as they claimed perhaps with a trace of hubris "Our results call into question the biological benefit of C60-OO in aging." - the benefit of c60oo on aging had not been established, they were here to shed light to that question, not to bring into question what was already being questioned: what is the role of c60oo on lifespan?. Baati produced data: we did this under these conditions, and produced that effect.
However, this slight of rhetorical hand signals to the reader they are disproving some established theory - not simply doing what they were doing, adding more data. More to the point they were not publishing basic data which creates tension with the study’s headlines and conclusions offering a hint of Baati-validation: this was no replication study.
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Nietzsche proclaimed one virtue to be better than two, for it is more of a knot to cling on to. Well, this study holds a second virtue, another knot, though a handy one. One rather like the first, smoothed out to avoid the friction of conflicting conclusions .
Take a look at the two survival charts. First Fig 3; only reported is the virtual identical median lifespan, not as mentioned that c60oo was shown to be life extending in mice over the effective within study control, the olive oil group - no reference to the two very distinct survival paths journeyed by the groups upon arrival at virtually identical medians - there was no lifespan shadowing.
When the olive group dropped down to a survivability of 55%, c60 was at the dizzying heights of 90% before collapsing onto the survivability percentage of the olive group. Prior to this was where the c60 lifespan gains were banked. This looks very strange, it seems c60 is doing as expected, something similar to Baati, then the cliff-edge. And in the end one olive oil mouse outlives all the c60 group.
The intervention looks to be initially protective, then harmful - a cumulatively hormetic relationship, perhaps - or the poor mice accidentally ingested some of Ichor's toxic light exposed c60oo!
This is a small sample size, and this could happen by chance - but could portend to something else, and would warrant a further look.
Move on to the second study: Fig 4, once again. What do we see this time? Almost identical behaviour: when the merged olive groups have a survivability of around 55%, with external controls below 60%, c60oo is once again at around 90% before dramatically collapsing onto those two groups with a shared survivability in the 50%'s. This is quite bizarre, yet unreported within the study.
This is likely very a highly confident effect, that it isn’t chance c60oo is initially lifespan-extending and subsequently lifespan-shortening. This analysis would need to be done, and the data made available. If as seems likely c60 is demonstrating a clear and distinct effect overall on life span then it attests to a likely complex relationship which this study is only partially exploring, as was the case with Baati only with very different outcomes.
There are other patterns of interest, though much weaker in confidence, once this perceived benefit is gone, the c60oo groups track the olive oil lifespans. The young c60oo group appears to salvage some lifespan benefit, hinting at the possibility that late earlier treatment, as per Baati, may have been beneficial - but this of course is quite possibly chance.
The conclusion for this study would have been that c60 in olive is life-extending in mice over olive oil controls, and these gains over olive oil harvested early in the study with c60oo offering no life extending gains to older olive oil mice, specifically the intervention becomes comparatively harmful late in life, and as such those early gains are not conserved into producing longer lived mice.
For Baati and Ichor c60 rodent studies to demonstrate c60oo to extend lifespan over olive oil "controls", where the former extends this relative lifespan by something like 60%, and the second to around 4% is suggestive of key study design differences and or the possibility of an hormetic relationship.
Breaking open the 4% life-extension gain provides strong evidence of an hormetic relationship between c60oo and rodent. One possibility is there being some negative cumulative effect of taking c60oo - perhaps as Turnbuckle and QuesforLife have indicated through different means the depletion of stem cells stores, though it might be expected that the mice starting c60 earlier would die more quickly were this true. There may be other cumulative negative effects of c60, the mention in one paper of c60 crystals becoming pro-oxidants. Another possibility is that the hormetic function changes with age.
The intervention may remain constant but the biological system will not - so the sweet spot of the hormetic curve may alter with age, and possibly, a dose that is optimal at one age, or even beneficial, may not be at another, possibly harmful. That the benefits of Baati could be sensitively calibrated should not be too surprising - which again is why it was imperative to replicate every facet of the study, before expanding into new territory.
Is there any evidence to support these hypotheses? Well, this in vitro study looking at olive oil's protective effect on dna was interesting.
https://pubmed.ncbi.nlm.nih.gov/16036359/
"Significant protection was observed in extracts from olive oil and olive mill waste water. However, above a concentration of 100 microg/ml olive oil extracts exerted DNA damaging effects by themselves in the absence of any H2O2. Extracts from olive leaves and olive fruits although protective, were also able to induce DNA damage by themselves."
Now if c60oo were to exaggerate the DNA-reparative and seemingly hormetic effect of olive oil, then we could very easily see c60oo switch from hero to villain. I would think it possible, at least, that one given dose of olive oil might protect against a certain degree of DNA damage, and be harmful to another i.e. the protective/harmful effects might not simply be a function of the olive oil dose, but a function of DNA damage, and so age.
The rapid death rate in the Ichor study at a certain point, should give pause for thought.
Another study looking at the potential harmful effects of oleic acid.
https://pubmed.ncbi.nlm.nih.gov/29730927/
Further strong supporting evidence for the sensitive calibration of c60oo, that the therapeutic window could be narrow, is evidenced in a study exploring the protective effects of thioacetamide (TAA) on the livers of Wistar rats. Take a look at these numbers on levels of catalase.
Controls - 72
TAA - 55
TAA + olive oil - 56.5
TAA+High concentration c60oo - 62.
TAA+Low concentration c60oo (20% of high dose) - 92.
C60oo at a high dose recovers 40% of the catalase lost to thioacetamide, but the low dose doesn't simply recover more, it rises to above 30% above controls.
There is no c60oo control for catalase. However in a study looking at the protective effect of c60oo on the pancreas of rats exposed to DMBA, catalase levels in the pancreas of rats receiving just c60oo (no DMBA) differed little from healthy control rats (no c60oo, no DMBA). Though c60oo partially recovered pancreas catalase levels in rats exposed DMBA (below)
https://pmc.ncbi.nlm...5826/figure/f3/
This provides strong supporting evidence to the unusual properties of c60oo - but suggests the potential narrowness of a therapeutic window.
In healthy controls c60oo appears not to change catalase levels in organ tissues, in damaged tissues depleted of catalase it recovers, further suggesting at some dose relationship it increases them 30% above controls.
Now this would seem quite easy to miss.
Were a lab looking to replicate this spiking effect in catalase they are going to need to get the dosing of c60oo and exposure to thioacetamide precisely right.
Further, were lab looking to find similar c60oo effects on catalase levels in the pancreas subject to DMBA, they are going to have to prepare to search extensively. Not knowing the narrowness of this sought-after behaviour, or even if it is there, they would have a number of parameters to explore: they could fix the DMBA and vary the c60oo exposure; do the reverse or vary both. That is quite a large testing space. What they are not going to do is choose one intervention and conclude there to be no effect.
The analogy with Baati/Ichor would be to put the former lab as the one doing the c60oo/Thiacetamide trial but without the high dose group. They would have made the seemingly extraordinary claim that c60oo not only diminished the catalase-loss caused by Thiocetamide but increased catalase 30% above controls.
Pantomime-Ichor steps in to confirm/debunk these claims by altering a few variables, “Well if it does this in the liver, then why not the pancreas with the catalase debilitating DMBA for these guessed at doses of c60oo in DMBA ?”.
Sure it could work, and it would be interesting, but it’s not replication.
And pantomime-Ichor lab finds that c60oo does not raise catalase above (healthy) control-levels in the pancreas DMBA intoxicated rats and openly brings into question the belief whether c60oo can increase catalase in the organs of poisoned rats above those of healthy control levels.
And critically, debunking pantomime-Ichor don’t deem it necessary to indicate c60oo partially recovering catalase levels above those in DMBA exposed rats (and so inferring that there is an effect of c60oo on catalase levels in DMBA exposed pancreas suggestive of a complex relationship). But for pantomime-ichor only two headlines matter - proven in pancreas, or disproven in liver. Then move on (and be sure and burn the baton.)
Now of course no lab could pursue this course given the Thioacetamide-c60oo lab chose two doses of c60oo demonstrating qualitatively different outcomes. An attempt to replicate these effects (and clearly not the study) in the pancreas would require a scattergun approach because there is too much uncertainty before concluding the effect was not reproducible. Failure to do so might raise some doubt over the validity of the c60-Thioacetamide liver-catalase results, but it would drastically fall short of discrediting the results - because the studies are different.
If a lab wishes to discredit the findings claimed, then they will have to replicate the study in all its specifics to show c60oo does not increase catalase levels above those of healthy controls in thioacetamide-exposed rats.
In short, Ichor managed the feat of appearing to attempt to recreate Baati, while introducing so many changes as to raise the chances of failure, and did not report the one result which was partially validating of Baati - and required explanation not forthcoming.
Edited by ambivalent, 17 May 2025 - 04:45 PM.
