120 replies to this topic

[smithx]

In case I haven't been clear about the samples, it should look like this:

001 - C60
002 - C60
003 - Placebo
004 - C60
005 - Placebo
006 - Placebo

in other words, when the samples are given to the test administrator, they should be completely randomized. It should not be one big block of C60 samples and another big block of placebos.

[maxwatt]

Standard tests of mice for vigor are roto-rod (how long a mouse can cling to a rotating rod) and swimming-to-exhaustion, or time on treadmill till exhaustion.
It is possible to hook an exercise wheel to a counter, so mice or groups of mice can be compared as to amount of physical activity the spontaneously indulge in.
Such might make possible comparative measures that would be a stand-in for retaining mitochondrial function with age.

I would also like to see a study that could tease apart the contribution of EVOO polyphenols; make a fullerend OO solution from a high polyphenol oil, and another from a low polyphenol oil, plus a control group, and possibly olive oil only control groups. My reasoning is that the olive-oil-only group showed almost 20% greater life span over controls, versus almost 50% improvement in OO + C60. Apparently the olive oil is not inert. Is it the oil itself, or the polyphenols therein that cause the effect? Do the bucky-balls act as a carrier by forming adjuncts with either polyphenols or oleic acid (a major component of olive oil?) which then get carried into the cells by fullerene molecules?

[AgeVivo]

rotarod

Edited by AgeVivo, 16 June 2012 - 02:16 PM.

[carbon]

If you guys can put together a serious study I would love to sponsor part of it by supplying C60 olive oil at cost.

www.carbon60oliveoil.com

Edited by carbon, 21 June 2012 - 07:05 AM.

[maxwatt]

rotarod

BOTH spellings are commonly used.

http://btc.bol.ucla.edu/rod.htm --
Improved rotorod performance and hyperactivity in mice deficient in a protein repair methyltransferase

RotoRod Protocol

ScienceDirect.com - Neuroscience Research - Rotorod ...


Rotarod Test for Mice and Rats

Influence of task parameters on rotarod performance and sensitivity ...

RotaRod | Biomodels

[brokenportal]

So who is in to do this study? Can we list some good candidates to go and contact to see about doing this too? Whose got the time and at least the bare credentials and know how to do this? We can gather estimates on amounts of money that people will do it for to consider too. I believe that Agevivo and wccaguy are in to do the study so far.

Edited by brokenportal, 24 June 2012 - 09:30 PM.

[carbon]

If the study is somewhat modest, say, 100 treated rats and 100 placebo rats, I can supply all of the C60/oo and placebo olive oil for free. I could send it all to a project coordinator who could send it out to the participants. I already produce a package for rat pet owners to try to reproduce this study. It consists of a 30 ml bottle and an applicator to feed it to your rats. In the study the rats got 24 one ml doses. I could make 200 of those packages, half with C60 and half just olive oil. Each bottle could be uniquely labeled with a random code. Then we could correlate the results to a spreadsheet. One benefit of this approach is that all of the C60 / placebo will be consistent. I have the capacity to produce this in a controlled, consistent way. I am not ISO 9000 yet, but I could provide process documentation for the batch.

[zorba990]

I'd really like to see hairless Mice and UV exposure in one study, one with topical application, one with internal low dose, internal high dose, both, and a control.
We should probably setup a webpage and a paypal/amazon account for those interested in contributing.

[carbon]

We should design a study that is friendly to the rats. We already expect good results for the rats because those are the results that we are trying to replicate. We should not do any of the following:

1) Cull rats for dissection.
2) Use gavage needles. The rats will drink this stuff. I provide blunt tip applicators in my kits.
3) Use I.P. injection. Yes the study did that for bio-distribution. We do not need to do that to replicate the longevity portion of the test.

One thing that is exciting about an open source / non-lab experiment is that we can provide a benefit to rats that are living in good, pet conditions. I have to agree with PETA that animal testing is a bad thing. We can turn it into a good thing by trying to give our pets the good results of this study, instead of feeding them C60 then chopping them up to see what happened. Also, I'm all for people taking the responsibility on themselves to experiment. A lot of people are reporting back results. We should try to collect that data.

[smithx]

I think it's great that Carbon is offering free test samples.

But since he is a vendor I propose the following, to avoid any claims anyone might make that there was tampering with the results to create a false positive effect:

- Carbon donates the C60 olive oil
- someone else with no vested interests produces the placebos (to avoid any claim that the placebos were poisoned or otherwise deleterious to the mice)
- either the person producing the placebos or someone else with no vested interests produces the labels and spreadsheet and keeps the key of which samples are C60 and which are placebo

[niner]

I would also like to see a study that could tease apart the contribution of EVOO polyphenols; make a fullerend OO solution from a high polyphenol oil, and another from a low polyphenol oil, plus a control group, and possibly olive oil only control groups. My reasoning is that the olive-oil-only group showed almost 20% greater life span over controls, versus almost 50% improvement in OO + C60. Apparently the olive oil is not inert. Is it the oil itself, or the polyphenols therein that cause the effect?

After seeing the paper about the effect of olive oil on mortality rates in the Spanish cohort of the EPIC population, and the dose-response relationship, I think a study on olive oil megadosing is warranted, with or without C60. Good olive oil vendors can supply the chemistry of a given batch, so one could compare the effects of differing polyphenol levels easily. While the C60 result stole the show, the olive oil arm of Baati could be called more important, in that it doesn't involve ingesting something that was unknown before 1985. This is not to say that I'm anti-C60; I'll probably be getting on that train myself one of these days- but the OO-only result isn't getting the attention it deserves.

[maxwatt]

I think it's great that Carbon is offering free test samples.

But since he is a vendor I propose the following, to avoid any claims anyone might make that there was tampering with the results to create a false positive effect:

- Carbon donates the C60 olive oil
- someone else with no vested interests produces the placebos (to avoid any claim that the placebos were poisoned or otherwise deleterious to the mice)
- either the person producing the placebos or someone else with no vested interests produces the labels and spreadsheet and keeps the key of which samples are C60 and which are placebo

And the keeper of the key is not the one who gives the samples to the tester.

[niner]

On the subject of recruiting pet owners to run the experiment, there are some major problems:

They may not know the age or species of their rats.

They are by definition emotionally involved with the animals, and might be tempted to give the active compound to both animals, perhaps by mixing the active and placebo, or by alternating them.

For the same reason, they might treat the animals differently, perhaps medicating one that gets sick.

If we send them two actives, or two placebos, instead of one of each, then the experiment becomes completely uncontrolled.

I think it would be better to start with a uniform population of animals, and keep them in one or several decent-quality facilities. We could do a lightweight version of this by recruiting people to run a mouse/rat group, and provide them all the equipment and animals they need for a reasonable number (5-10?) of rodents. I would rather have the people running this be in it for the science, as opposed to rat-o-philia. We want people who like the animals enough to care for them well, but not so much that they might blow the experiment to save one of their babies.

[smithx]

I think it's great that Carbon is offering free test samples.

But since he is a vendor I propose the following, to avoid any claims anyone might make that there was tampering with the results to create a false positive effect:

- Carbon donates the C60 olive oil
- someone else with no vested interests produces the placebos (to avoid any claim that the placebos were poisoned or otherwise deleterious to the mice)
- either the person producing the placebos or someone else with no vested interests produces the labels and spreadsheet and keeps the key of which samples are C60 and which are placebo

And the keeper of the key is not the one who gives the samples to the tester.

Exactly.

That part was covered in my previous suggested protocol, but it's good to highlight it.

[smithx]

For the same reason, they might treat the animals differently, perhaps medicating one that gets sick.

This is exactly why we can't send anyone both a placebo and a C60 sample.

If they know they have one of each, they may be tempted to try to figure out which is which, and give the C60 to their favored or sickly animal.

There is also the chance that samples would be mixed up and given to the wrong animals, if one pet owner got both types. That's why I suggest that each pet owner get either placebo or C60.

If we are going to have pet owners treat multiple animals, we would produce sets of different sized bottles:
- Single animal bottles
- Two animal bottles
- Three animal bottles
- etc.

In each case, there would be an equal number of C60 and of placebo bottles, randomly labeled, with a key spreadsheet.

In this way, we will be able to have some pet owners treat multiple pets, but make sure that there's no chance of mixups, accidental or otherwise.

Edited by smithx, 26 June 2012 - 10:09 PM.

[niner]

I don't think it would be publishable without controls. It would also stand a good chance of not telling us what we need to know. I really think we need a controlled experiment.

[smithx]

I'm not sure what you're thinking of.

The placebo animals would be the controls. There would be an equal number of placebo vs C60 animals.

That seems controlled to me.

The only issue that comes to mind is the ages of each group, but with a large enough sample size, the ages should be distributed roughly evenly between groups, just through random assignment.

[Turnbuckle]

I don't think it would be publishable without controls. It would also stand a good chance of not telling us what we need to know. I really think we need a controlled experiment.

It's not going to be publishable without credentials.

[niner]

I'm not sure what you're thinking of.

The placebo animals would be the controls. There would be an equal number of placebo vs C60 animals.

Not very good ones, since they would be in different environments, with different levels of care, and there would be problem of unknown or mis-assigned ages and breeds. With a big enough sample size, we could still get a signal from the data; it's just a lot noisier than it would be in an animal facility. That means we'd need a much larger sample size for the same information. I think it's going to be hard to get the number of people we'd need for decent statistics.

[Mind]

On the subject of recruiting pet owners to run the experiment, there are some major problems:

They may not know the age or species of their rats.

They are by definition emotionally involved with the animals, and might be tempted to give the active compound to both animals, perhaps by mixing the active and placebo, or by alternating them.

For the same reason, they might treat the animals differently, perhaps medicating one that gets sick.

If we send them two actives, or two placebos, instead of one of each, then the experiment becomes completely uncontrolled.

I think it would be better to start with a uniform population of animals, and keep them in one or several decent-quality facilities. We could do a lightweight version of this by recruiting people to run a mouse/rat group, and provide them all the equipment and animals they need for a reasonable number (5-10?) of rodents. I would rather have the people running this be in it for the science, as opposed to rat-o-philia. We want people who like the animals enough to care for them well, but not so much that they might blow the experiment to save one of their babies.

Because the lifespan effect of the C60 + Olive Oil mixture is supposed to be so extreme, these objections matter little for the "pet owner experiment" (IMO). We can fund a different controlled study in a lab somewhere in the world and get robust results and attempt to get it published.

The main thing is that the owners keep a log of their feeding schedule and care and that they have a reasonable estimation of the age of the rats when they start the experiment. A bonus is if they would share pictures and/or video of the animals. I view the "pet owner experiment" as something cheap and fast to give us a solid next level of evidence. The lifespan extension is supposed to be so extreme that we can deal with some uncertainty and still see a noticeable result. If all of these rats are living many months or years (as Baati would suggest) beyond normal then the pet owners should see noticeable results. If, on average, all of the rats are keeling over pretty much on schedule, then we have an inkling that there might have been something wrong with the original experiment. Agevivo has already started a small sample experiment - bravo! If we could get a few dozen rat owners to start feeding them the mixture, how much would that cost? We only need to order and ship the stuff.

Edited by Mind, 27 June 2012 - 09:48 PM.

[AgeVivo]

For the same reason, they might treat the animals differently, perhaps medicating one that gets sick.

This is exactly why we can't send anyone both a placebo and a C60 sample.

If they know they have one of each, they may be tempted to try to figure out which is which, and give the C60 to their favored or sickly animal.

I don't think these are much of issues

There is also the chance that samples would be mixed up and given to the wrong animals, if one pet owner got both types. That's why I suggest that each pet owner get either placebo or C60.

This is the biggest issue I see in what has been discussed on this page, concerning non extreme life extensions at home. Basically it means that either you are able to find say 10 pet owners who are *VERY* careful, or you find at least *20* pet owners having one cage. But perhaps it is feasible after all -- perhaps it just needs to be tried.

Because the lifespan effect of the C60 + Olive Oil mixture is supposed to be so extreme, these objections matter little for the "pet owner experiment" (IMO). We can fund a different controlled study in a lab somewhere in the world and get robust results and attempt to get it published.

The main thing is that the owners keep a log of their feeding schedule and care and that they have a reasonable estimation of the age of the rats when they start the experiment. A bonus is if they would share pictures and/or video of the animals. I view the "pet owner experiment" as something cheap and fast to give us a solid next level of evidence. The lifespan extension is supposed to be so extreme that we can deal with some uncertainty and still see a noticeable result. If all of these rats are living many months or years (as Baati would suggest) beyond normal then the pet owners should see noticeable results. If, on average, all of the rats are keeling over pretty much on schedule, then we have an inkling that there might have been something wrong with the original experiment. Agevivo has already started a small sample experiment - bravo! If we could get a few dozen rat owners to start feeding them the mixture, how much would that cost? We only need to order and ship the stuff.

Thank you, I fully agree on all this. Moreover, having "normal people" give something supposed to increase the lifespan of their pets has a extremely strong mindshift impact on the perception of desirability and feasibility of life extension.

Edited by AgeVivo, 29 June 2012 - 07:20 PM.

[Mind]

So I am thinking about making a post soon in the pet forums about the rat experiment. Trying for too much scientific rigor in this part of the investigation (pet owners) would add too much complexity and too much room for failure. Giving the rats the C60oo mixture at intervals corresponding to the Baati experiment should be fairly easy for a dedicated pet owner.

I will post in the pet forum about the original C60oo experiment and link to AgeVivo's ongoing experiment. I suspect the more people might be willing to try it if they see someone else doing it and the pets have not died, but seem quite healthy.

Screening Pet Owners:
Those who have the best estimate or documentation of the age and species of their pets would get the highest consideration.
Those who agree to provide pictures/data/video/log of their home experiment in the Longecity forums would get higher consideration.

Legal:
They would have to sign an agreement removing Longecity from any legal/monetary liability.

Mechanics:
We would have to find a good source of the elixer and method of shipping. We might need to ship a measuring device as well so the owners could dose the correct amount.

[Logic]

Because the lifespan effect of the C60 + Olive Oil mixture is supposed to be so extreme, these objections matter little for the "pet owner experiment" (IMO). We can fund a different controlled study in a lab somewhere in the world and get robust results and attempt to get it published.

The main thing is that the owners keep a log of their feeding schedule and care and that they have a reasonable estimation of the age of the rats when they start the experiment. A bonus is if they would share pictures and/or video of the animals. I view the "pet owner experiment" as something cheap and fast to give us a solid next level of evidence. The lifespan extension is supposed to be so extreme that we can deal with some uncertainty and still see a noticeable result. If all of these rats are living many months or years (as Baati would suggest) beyond normal then the pet owners should see noticeable results. If, on average, all of the rats are keeling over pretty much on schedule, then we have an inkling that there might have been something wrong with the original experiment. Agevivo has already started a small sample experiment - bravo! If we could get a few dozen rat owners to start feeding them the mixture, how much would that cost? We only need to order and ship the stuff.

+1!

This is just a quick and cheap way of making sure the stuff works.
It should also help spread the 'word' very well.

Edited by Logic, 03 July 2012 - 12:17 AM.

[smithx]

So I am thinking about making a post soon in the pet forums about the rat experiment.

What's your thinking about the points in my suggested protocol?

Have you picked an online survey tool?

[Mind]

So I am thinking about making a post soon in the pet forums about the rat experiment.

What's your thinking about the points in my suggested protocol?

Have you picked an online survey tool?

I will review these issues before moving forward. Thanks for checking in again smithx.

[Brainbox]

I like this initiative! I'm not around to much lately and must say that I'm very very pleased. Unfortunately I do not have the time to keep some animals myself, but will try to find a way to participate in this.

One suggestion: Lets obtain the olive oil for the control group from a neutral and different source from which we get the c60/oo. If we design a good randomized double blind feeding and measuring scheme, than we should also avoid the culprit of possible (unintended) skewing by manipulating the control oil negatively. I'm not implying to mistrust carbon, but for the sake of a good study design this could be very essential. To avoid offset due to the use of different types of olive oil, we could obtain the control group oil from the same source and use the same brand / type, but I have no idea if this is important.

Edit: missed these postings before.

I think it's great that Carbon is offering free test samples.

But since he is a vendor I propose the following, to avoid any claims anyone might make that there was tampering with the results to create a false positive effect:

- Carbon donates the C60 olive oil
- someone else with no vested interests produces the placebos (to avoid any claim that the placebos were poisoned or otherwise deleterious to the mice)
- either the person producing the placebos or someone else with no vested interests produces the labels and spreadsheet and keeps the key of which samples are C60 and which are placebo

And the keeper of the key is not the one who gives the samples to the tester.

What would make it more safe? Containers that are sealed by the manufacturers? How are tests that have a certain risk for manipulation (to some extend all tests are) run professionally?

Edited by Brainbox, 04 July 2012 - 08:40 PM.

[smithx]

So I am thinking about making a post soon in the pet forums about the rat experiment.

What's your thinking about the points in my suggested protocol?

Have you picked an online survey tool?

I will review these issues before moving forward. Thanks for checking in again smithx.

Please let me know if any point seems unclear or unnecessary, or if you disagree with any of them. I have done a lot of scientific studies, and every point has a reason, so I will be happy to provide additional information if needed.

[niner]

One suggestion: Lets obtain the olive oil for the control group from a neutral and different source from which we get the c60/oo. If we design a good randomized double blind feeding and measuring scheme, than we should also avoid the culprit of possible (unintended) skewing by manipulating the control oil negatively. I'm not implying to mistrust carbon, but for the sake of a good study design this could be very essential. To avoid offset due to the use of different types of olive oil, we could obtain the control group oil from the same source and use the same brand / type, but I have no idea if this is important.

I think it's important to use identical oil for the solution and the control, otherwise it's not a real control. I see no reason to introduce another source of randomness if we can possibly help it. In fact, the control oil should be from the same batch as the C60 solution oil, since olive oil chemistry varies a lot. We could always supply the oil to carbon and have him brew a batch, if he's willing to do that.

[carbon]

Sure, I could do that. It is because of these issues that I originally suggested bottling both the C60 and non C60 oil, but however you guys design the study is up to you. The oil that I have been using is working out great. One thing about the oil though. Currently, I am getting oil from the Mediterranean, but I live in the best olive producing area in the United States - the North central valley of California. The olive oil harvest is coming up in a few months, and I want to switch to a local supplier. I think it would be awesome to see the olives get pressed and then immediately add C60 and start mixing. That way I will know for sure the source and quality of the oil, and the C60 will immediately start preserving it. This should make the best possible quality product, and once I have it all dialed in with a local organic olive farmer I would like to stick with that. But if you guys want to source some oil, I would be happy to mix a batch.

[AgeVivo]

I live in the best olive producing area in the United States - the North central valley of California (...) if you guys want to source some oil, I would be happy to mix a batch.

Here is a proposed way of functioning:

- Someone sends the oil to Carbon; say for example s123 (as he was proposing to handle the mixtures for Mprize at home)
- Carbon makes the mixture with C60 and send it to s123
- s123 sends labelled tubes to pet owners.

The pet owners either receive oil or C60+oil: they don't know. s123 is the only one to know. Perhaps we need to secure it a little more, for example s123 sends "who receives what" to some mailing address and the letters will be opened in the future, under a video-camera.