Let's focus on the issue nobody has really solved - how do we know we have the "right c60?" - we need to establish this first, and these tests can lead to designs of better tests that rule out this problem.
Also the pilot study did confirm the life extension properties (it just used different c60 oil), I'm not sure how much the mice strain's life span was considered in statistical calculations, but there must be some kind of error correction right? e.g. I am assuming the mice are dying before / or close to the minimum age in the current 3 tests.
Was there a particular reason wistar rats were not used?
Agreed on your first point. This must be established first.
Wistar rats were not used in our study because we only use mice in our laboratory (though we expect to have rats in the near future) and because of the cost and time expense. C57BL/6 male mice are the only model that can be purchased pre-aged, which knocks about a year off the study duration. That is largely why this model was chosen.
And if I can add my questions to Kmoody:
1) What was the rationale behind picking the current 4mg/kg weekly dosage (both in the AML and the current study)? Why has not the more effective 8mg/kg dose used in the AML study been espoused?
2) Given the strong suspicion of the particular C60oo product being the major cause for the currently observed accelerated mortality, are you planning to replace the C60oo source or you will continue running the study with the original C60oo?
3) Is it logical to conclude if these very cancer-prone mice are not developing cancer then a non-genetically crippled mouse would fare better or that would be too big of a leap in thought?
1. I might have mis-typed that. We are doing 8mg/kg in the AML study. We chose the 4mg/kg because this is similar to what Baati chose. Since we were using a dietary form of administration we opted not to do a loading dose but did not slow or stop administration later as Baati did. We keep an approximate 4mg/kg/week going throughout.
2. No. We will complete at least two of the three studies as they are. For the lifespan study, we may choose to terminate the C60oo arms and restart it using our formulation. This has not yet been decided however, and is pending more data.
3. Too big of a leap in thought. But note that with the off-the-shelf C60oo we are seeing an INCREASE in cancer burden in our AML model and our lifespan study mice (a different strain) are not all cancer free.
One thing is sure: sonificated C60oo is a whole different compound and should not be used in research for our purposes.
I agree this is likely. Which is why I am not suggesting C60oo does not have positive effects (particularly since I saw such a dramatic positive effect in my pilot study). Rather, we think that the C60oo purchased off-the-shelf from the vendor may have been prepared in a manner inconsistent with our (and Baati's) prep.
Kmoody, could you confirm that the control group is being fed plain OO? If this is not the case, do you agree that a reasonable alternative hypothesis for the observed increase in mortality in the C60OO group is that the fat is bad for these mice?
For the AML there is an olive oil only control. For the lifespan study we do have an indirect control for olive oil.
Group 1: C60oo only
Group 2: Olive oil + Proprietary Formulation
Group 3: C60oo + Proprietary Formulation
Groups 1 and 3 are trending worse than group 2.
Despite all the hype, no one really knows what those rats were given and what is having the positive effects. There is only speculation until a properly manufactured product shows efficacy or a mechanism of action is clearly determined.
But haven't you plunged into testing on animals using a product prepared by a method unknown to you (at the time) and highly problematical? People here saw the SES/oil product as suspect some time ago. Anything prepared with heat, high shear or ultrasonic energy has to be suspect. And the same goes for using an oil product that is years old as one person here did. So any data generated that indicates a problem can't be said to be a problem with C60/EVOO per se, but with C60/EVOO prepared by a certain method or containing some level of rancidity.
Yes. This is precisely why I want to get away from research grade studies and move into FDA compliant studies. It eliminates these variables. What seems to not be appreciated here is "plunging into testing... using a product prepared by a method unknown to you" is how most research is done in small companies and in many universities.
If you did this study at a university, 9 times out of 10 you would have bought the stuff off the shelf and believed the vendor's product specifications and that this product was manufactured the same way as Baati's. In fact, we were never paid nor even asked to do C60oo quality control in the scope of any of our studies. This is something we did on our own in an attempt to ensure the quality of our science, based in part, by quality assurance issues raised on this forum (so kudos guys for making sure we were pointed in the right direction). 
So yes, I do not believe the problems we see have anything to do with C60oo. I believe the problems have to do with a product labeled C60oo from one particular vendor. I feel the most efficient way to address this problem is to manufacture and characterize a C60oo product under FDA compliant GMP standards. Then we will KNOW what we are using. We will KNOW the shelf-life, the breakdown products, etc. GMP is an exceptionally high burden of quality assurance in the manufacturing process. You can put anything in a box that is "research grade" and almost always get away with it.
I believe he said that he was actually worried the quality of the C60 he was preparing wouldn't be as good as something that came from the actual manufacturer SES. He simply assumed he was actually using higher quality C60 when he isn't - well we don't know for sure yet what happened.
Yeah I read the C60 thread some people did raise these points, but they are buried as the c60 thread is like 60 pages or something so he probably never read those comments, no doubt he is getting a lot of useful feedback for a better design in the future.
Correct on both accounts, though some folks on this forum were quick to point out the C60oo concerns which is partially why we were quick to test out our vendor's product as soon as we were able to.
