After digging around over the last few days, I believe there to be likely a very different explanation, at least in a signifcant part, as to the benefits of this protocol, in particular the explanation of the reversal of DNA damage recorded by Turnbuckle through DNA tests.
There seems quite strong evidence to imply that the reversal of DNA damage results from c60 regulating/inducing hormesis in Hydrogen Peroxide. Two fairly recent c60/Wistar papers would seem to point indirectly to this conclusion or to some similar explanation.
In a toxicology study on the effects of c60oo intervention on Thioacetamide (TAA) exposed rats, the low dose c60oo group not only protected against a more than 200% increase in the comet tail - a marker of DNA damage - but remarkably conferred a 30% reduction in tail length over controls. In line with Turnbuckle's observations that low doses are more beneficial, the high dose group (5x) all but wiped out the damage but still retrieved a comet tail greater than those of controls. The inevitable question is to wonder whether c60oo would reduce the comet tail without the burden DNA damaging TAA, without the burden of the DNA damaging TAA. Unfortunately, there was no C60oo group, nor H2o2 measurements.
I would guess though it quite unlikely that c60oo would repair DNA, reduce the tail "on its own", as it were. For one Turnbuckle had over the years taken c60 as have others and though DNA methylation numbers haven't been over-reported, Turnbuckle only witnessed a significant decrease when starting this protocol despite taking c60oo for years at low doses. There have also been failed (mouse) longevity studies. And many people have often noticed an initial benefit, then things drop off under constant use. The other c60 Wistar rat paper, on measuring the intervention on a Huntington Disease (HD) model of C60 in DMSO rather supports that belief. Unlike, the TAA study, there is a lone c60 group to compare with controls. When looking at the array of metrics c60 appears to do absolutely nothing - they could almost be interchangeable with controls, next to no difference on H2o2 or SOD2 levels. Yet when present in the disease states c60 has an enormous impact, for example, pulling in H2o2 concetrations to controls, recovering the GSH cycle, resulting in hugely improved outcomes over the short trial.
C60 for the most part seems to quietly regulate, maintaining homestasis, until something gets of line. The stress-responses to fasting, exercise and some toxins too have been long shown as beneficial at the right level. In the TAA study, remarkably, c60 at the low dose, turns around this extremely harmful concetration of TAA into a signifcant benefit - to the comet tail at least - reducing DNA damage. C60 seems to make it the "right dose".
Given, the apparent lack of this benefit amongst c60oo users, yet clear evidence of it from Turnbuckle, and quite possibly too with the Baati rats, this has to be where an explanation and a safe strategy is sought from. And there seems to be one for both Turnbuckle, Baati and TAA/C60oo. And it is Hydrogen peroxide, H2o2.
In another study on Wistar rats, TAA was shown to signifcantly increase H2o2. In Turnbuckle's' protocol, H2o2 raising palmitric acid (in vitro) is present in stearic acid and perhaps H2o2 is triggered by nicotinamide too, and in the Baati study, as mentioned by Turnbuckle, the rats were fasted, and fasting has been shown in a number of animal models to signifcantly raise H2o2 (in the liver).
So why H2o2? Well, in one study H2o2 was shown to reduce lifespan in all groups of housefiles, except one, the 10% of max H2o2 dose group, which in fact extended lifespan. And H2o2 has been shown to cause DNA damage, as measured by comet assays, while of course too TAA caused DNA damage, as well as known to raise H2o2.
From this paper:
"And Increased hydrogen peroxide in catalase-deficient cells extends chronological lifespan despite parallel increases in oxidative damage. These findings establish a role for hormesis effects of hydrogen peroxide in promoting longevity that have broad implications for understanding aging and age-related diseases"
In the c60/DMSO HD model there is signifcant recovery of raised H2o2 in both the mitochondria of muscle and brain cells. And too there appear clear hormetic effects of H2o2 in extending lifespan, a study demonstrating a c60 induced hormetic effect for the DNA damaging TAA, which in turn has been shown to increase the DNA damaging H2o2.
So there seems astrong possibility that C60 is regulating H2o2 to induce a DNA repairing, life extending hormetic effect. Ordinarily, presumably, endogenous H2o2 is not enough, so some level of exogenous H2o2 needs to be hormetically regulated - except perhaps during fasting, as might be per Baati.
Further and finally:
"This effect indicates that control of hydrogen peroxide production regulates the mitochondrial respiration preventing the insulin resistance in skeletal muscle cells by a mechanism associated with CREB phosphorylation and β-oxidation of fatty acids."
So this might explain some of the immediate mitochondrial-related benefits many of us have experienced, C60 may well regulate H2o2 production and so in turn mitochondrial resporation.
Most links have not been included but are found in the more extensive post on the forum. These papers seem important and haven't been posted, and may not be spotted on the forum by those just following the thread.
This doesn't negate the protocol - it has induced the benefits in Turnbuckle and others - but it may mean that all of the benefits are not explainable through the description of this protocol and so perhaps improved. Certainly it seems quite likely that the C60/TAA provides an explanation for Turnbuckle's lowering of DNA methylation guven c60's dramatic effect on comet tail in this study. And H2o2 hormesis is a strong candidate and present in the protocol with palmitric acid. Also, it might encourage a more cautious approach around levels of c60 and stearic acid/palmitric acid. Perhaps too, it might partly explain the variance in methylation ages - assuming - comet tail and DNA methylation correlate - fluxxing levels of H2o2 and perhaps the timing of c60 as well is the pre-existing levels of c60 when H2o2 is raised.
It might too provide a credible explanation for Baati given the c60oo was stopped years before death, if DNA damage was held back itself for a couple of years.
Edited by ambivalent, 18 December 2022 - 10:37 PM.
