120 replies to this topic

[Mind]

We would have to explain ahead of time to the rat owners that there will be a placebo involved. I am confident we can some some of them that understand the necessity of placebo control.

[Mind]

The lady who "runs" the Rat Fan Club, is very interested in our concept for trying some citizen science. She is currently reading the Baati study. She has recommended somewhat experimental substances to rat owners in the past. Things are looking positive thus far. All I am hoping is that she will help us reach out to rat owners in order to gauge the interest in a possible study.

[Raphy]

Great news, thanks

[Logic]

Mind:

AgeVivo said that " We will soon start mprize at home with c60 (pet owners have answered positively..."
in the 3 mice at home thread.

I assume you have had some positive feedback from the rat fan club?

[Mind]

I have only talked to the owner of the Rat Fan Club website. I am going to make more inquiries now that we have our samples/methods/team set-up. Of course, if anyone would like to buy a couple of Wistar rats and raise them at home, please speak up. It is of course a long-term commitment to care for the animals (5 years...if Baati is a valid study, 2-3 years if your pets gets placebo). You would get the C60oo for free.

[niner]

If the goal is to duplicate Baati exactly, then people should be aware that Baati's rats got injected with C60-oo through a tube that was inserted down their throat. This sounds like something that takes a lot of animal handling expertise. I don't actually think that we need to attempt an exact duplication, particularly considering the other issues with the pet owner scheme. We just need to show significant life extension in mammals, preferably a shorter-lived species so that we can get results faster. (Which I realize is at odds with designing an experiment that is relevant to long-lived mammals like us.)

[Mind]

If the goal is to duplicate Baati exactly, then people should be aware that Baati's rats got injected with C60-oo through a tube that was inserted down their throat. This sounds like something that takes a lot of animal handling expertise. I don't actually think that we need to attempt an exact duplication, particularly considering the other issues with the pet owner scheme. We just need to show significant life extension in mammals, preferably a shorter-lived species so that we can get results faster. (Which I realize is at odds with designing an experiment that is relevant to long-lived mammals like us.)

We would like to get the approximate dosage in the pet rats, without the gavage process. Obviously, we cannot replicate Baati exactly with pet owners at home. The attempt here is to balance community building (open science) with quality research. We are hoping to get somewhat robust data with a larger sample.

[Logic]

Thx for the update mind.
Shout if you need a hand contacting people or anything.

[AgeVivo]

We would like to get the approximate dosage in the pet rats, without the gavage process.

Actually it will be quite precise and I would say better than the gavage process. Gavage cause a lot of stress for the animal (a lot). Here, no stress: you change the cage on Friday evenings for example. You don't give food right away: The next day they are hungry (a little like you after the night); you prepare as many pieces of bread as you have pets; you put a given number of solution drops on each of them. One after the other one, you put one mouse only in the cage with one piece of bread. You wait until it is eaten (fast because the animal is a little hungry) and you go to the next animal. Simple, precise, no stress. When done, you put food in the cage.

[Mind]

We would like to get the approximate dosage in the pet rats, without the gavage process.

Actually it will be quite precise and I would say better than the gavage process. Gavage cause a lot of stress for the animal (a lot). Here, no stress: you change the cage on Friday evenings for example. You don't give food right away: The next day they are hungry (a little like you after the night); you prepare as many pieces of bread as you have pets; you put a given number of solution drops on each of them. One after the other one, you put one mouse only in the cage with one piece of bread. You wait until it is eaten (fast because the animal is a little hungry) and you go to the next animal. Simple, precise, no stress. When done, you put food in the cage.

This will be our feeding protocol. Thanks AgeVivo.

[Shannon Vyff]

We have had rats on and off for years and would be interested in doing this, you can add us to the list.

[Mind]

Wonderful. Thanks Shannon!

[smithx]

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.

This again is not optimal because you have one person creating the samples and the same person deciding who gets what.

A better way to do it is:

- Person A supplies a set of C60 samples
- Person B supplies a set of placebo samples
- Person C randomizes the numbers and puts the randomized numbers on each sample. So sample 1 may be either C60 or placebo, sample 2 may be either C60 or placebo, etc. Only the key lets us know which is which.
- The key should be put into an encrypted file and distributed to several people to insure that it isn't tampered with. The password to the file should be kept secure in one place.

- After this process is complete, Person D gets a box of samples which are completely randomized. This person has no way of knowing which is which.
- Person D sends the samples to the test subjects, monitors them and collates the results

- The experiment should have a defined time limit longer than the expected lifespan of any of the mice, or end when all the mice are dead, whichever comes first
- After the end of the experiment, the data should be analyzed and published
- Only after this step should the key be revealed, so that we can see if the lifespan of the mice was correlated in any way with whether they got placebo or C60.
- The fact that the data is analyzed before knowing the key, and the fact that they key was held by several people and therefore hasn't been tampered with, will prove the validity of the results.

[niner]

This again is not optimal because you have one person creating the samples and the same person deciding who gets what.

A better way to do it is:

- Person A supplies a set of C60 samples
- Person B supplies a set of placebo samples
- Person C randomizes the numbers and puts the randomized numbers on each sample. So sample 1 may be either C60 or placebo, sample 2 may be either C60 or placebo, etc. Only the key lets us know which is which.
- The key should be put into an encrypted file and distributed to several people to insure that it isn't tampered with. The password to the file should be kept secure in one place.

- After this process is complete, Person D gets a box of samples which are completely randomized. This person has no way of knowing which is which.
- Person D sends the samples to the test subjects, monitors them and collates the results

- The experiment should have a defined time limit longer than the expected lifespan of any of the mice, or end when all the mice are dead, whichever comes first
- After the end of the experiment, the data should be analyzed and published
- Only after this step should the key be revealed, so that we can see if the lifespan of the mice was correlated in any way with whether they got placebo or C60.
- The fact that the data is analyzed before knowing the key, and the fact that they key was held by several people and therefore hasn't been tampered with, will prove the validity of the results.

I don't see why persons A and B shouldn't be the same person. In fact, a single person would ensure that the oil was handled identically, thus making it a better control. If the samples and controls are really indistinguishable, then the A/B person could also fulfill the role of person D. Did you really mean that the code should not be broken until after the paper is published? I don't see how that works. I think the whole idea of publication is questionable, given the random set of animals, environments, and animal handlers. I'm afraid that peer reviewers are going to have a hard time taking that seriously. Maybe the ultra high-security blinding is overkill, given the randomness in the experiment.

Come to think of it, if we are using an olive oil control, then we are not going to do the blank control, which was water in Baati, and would presumably be a piece of bread with nothing on it in our case. Considering the large effect that Baati saw from olive oil, and the greater degree of noise in our data, we could be setting ourselves up for a result that's lost in the noise. We could perhaps fix this by using corn oil, or even a "lite" olive oil as the control.

[smithx]

I don't see why persons A and B shouldn't be the same person. In fact, a single person would ensure that the oil was handled identically, thus making it a better control. If the samples and controls are really indistinguishable, then the A/B person could also fulfill the role of person D. Did you really mean that the code should not be broken until after the paper is published? I don't see how that works. I think the whole idea of publication is questionable, given the random set of animals, environments, and animal handlers. I'm afraid that peer reviewers are going to have a hard time taking that seriously. Maybe the ultra high-security blinding is overkill, given the randomness in the experiment.

Come to think of it, if we are using an olive oil control, then we are not going to do the blank control, which was water in Baati, and would presumably be a piece of bread with nothing on it in our case. Considering the large effect that Baati saw from olive oil, and the greater degree of noise in our data, we could be setting ourselves up for a result that's lost in the noise. We could perhaps fix this by using corn oil, or even a "lite" olive oil as the control.

The reason I have them separate is because A is likely to be a vendor of C60 products, and they should not touch the controls because someone could claim they have a motivation for poisoning them or otherwise adulterating them.

By publication, I mean, publication here. If the data is published online before the key is published, then there's zero chance that anyone fudged the results.

[smithx]

People viewing this thread may not be aware that the actual planning for this study is being done elsewhere, in this thread:
http://www.longecity...munity-science/

Everyone who would like to have input into how the study is actually conducted should post there instead!

[clairvoyant]

Simply replicating Baati et al. would be an achievement. That would take up to four years, however, which is kind of long. I would hope that Baati's group already has a second study in the wings that is well under way. It has probably been a couple years since they had the earliest indications of unusual lifespans, so that wouldn't be unheard of. Because of the time involved in doing a rat lifespan study, not to mention the expense, it might be worth thinking about other shorter-lived species.

It would be good to nail down exactly what you get when you mix C60 with olive oil. Is it C60 dissolved in oil, a reaction product in which C60 is covalently bound to a long chain fatty acid ester, or some mixture of these? This is just an analytical chemistry problem, and it probably wouldn't take long in the appropriate lab.

A smaller, cheaper, shorter-lived species would allow us to ask questions like what is the optimal oil or what is the optimal dosing schedule.

Using radio-labeled C60, we could look at tissue localization and the kinetics of clearance. The effects that extend far beyond the last dose suggest an exceptionally long residence time; it would be nice to confirm or refute that hypothesis. This could be done more quickly than a lifespan study.

All great thoughts. I take their import to be that several smaller projects would move us forward faster than a single project aimed at replicating Baati et al even if a few other findings were included.

• From a post by Anthony Loera in that other thread, I take it that he may have contacts who know people on the Baati et al team. Perhaps, we could get Anthony to find out what they're up to.

• I especially like the idea of a small, initial project to determine what happens when C60 is mixed with Olive Oil. My initial thought is that the lab which determined that GPR40 "directly mediates" Olive Oil-"induced secretion of cholecystokinin" (CCK) would be a good candidate lab to do this. Perhaps, because of their previous experience, they could resolve the C60/Olive Oil bonding question as well as the question of what happens to that bond following CCK secretion.

• Getting at the explanation for the effects that extend far beyond the last dose would be a profound achievement. As I understand it, there are at least two hypotheses for those long lasting effects.

the C60s have an exceptionally long residence time in tissue (although Baati et al say the time for clearance was measured in "tens of hours")

from a Turnbuckle post, the C60 "acts to strip methyl groups from the mitochondrial DNA"

I don't know enough to know whether these are the only two hypotheses which might explain the long lasting effects. Are there others?

• Do you have a thought about what animal(s) might be appropriate to try the formulation with rather than rats?

These are great ideas niner. I like them even more now that I've slept on them before posting this reply!

I would like to comment the long lasting effect of C60 AFTER the discontinuation of its administration. Possible cause for this might be that C60 repairs lipid peroxides into normal ones (already peroxided fat tissues(including membranes)) and the animals becomes younger. After stopping of C60 they have extra "time" to get old.

I have already bought 2 grams C60 99.95% for $45 a gram plus $90 for shipping=$170. I am waiting to receive them. I am going to dose myself with 10 MICRO grams first dose and raise it by doubling it, so that it can reach 0.1 -1 MILLIGRAMS per day. If anything occurs I will stop at lower doses, because I have read that 10mg have been taken by humans. Nevertheless I am afraid of possible long lasting deleterious effect but more of old age and death.
The first signs of aging started to appear on me.
My body weight is 70 kg, 44 year old, 1.70 meter high, low body fat with muscles.
My mother is afraid for me not to die. Please, wish me success. I will report.
I think(hope) that c60 not only stop aging but rejuvenate, too. Thus, it will be good to have a tread in this site called: human testimonials.

[lifequester]

I would like to comment the long lasting effect of C60 AFTER the discontinuation of its administration. Possible cause for this might be that C60 repairs lipid peroxides into normal ones (already peroxided fat tissues(including membranes)) and the animals becomes younger. After stopping of C60 they have extra "time" to get old.

I have already bought 2 grams C60 99.95% for $45 a gram plus $90 for shipping=$170. I am waiting to receive them. I am going to dose myself with 10 MICRO grams first dose and raise it by doubling it, so that it can reach 0.1 -1 MILLIGRAMS per day. If anything occurs I will stop at lower doses, because I have read that 10mg have been taken by humans. Nevertheless I am afraid of possible long lasting deleterious effect but more of old age and death.
The first signs of aging started to appear on me.
My body weight is 70 kg, 44 year old, 1.70 meter high, low body fat with muscles.
My mother is afraid for me not to die. Please, wish me success. I will report.
I think(hope) that c60 not only stop aging but rejuvenate, too. Thus, it will be good to have a tread in this site called: human testimonials.

since starting this personal trial last october, do you have any updates to report clairvoyant? such as, are you still taking it? and if so, what dosage are you on at present - are there any noticable effects to report?

[clairvoyant]

Hello, liferequester,

I took C60 in olive oil. Dose of 0.1 micrograms was too big to me. It may be because I am healthy and don't drink. The adverse effect was hyper activity, heat and energy. Sleep disruption, too.
I posted my experience in the forum "C60 at home". Now I am taking C60-oo in pico gram range on a weekly base. I have many health benefits.
For example skin, endurance, strength, sense of hearing, nerve reaction time, recuperation time improvements and others. If you look like what is in the picture, I recommend you take it immediately. Of course, if you want to live long and healthy. Because, in my country Bulgaria, people do not, and take alcohol and cigarettes ad lib.
Find you dosage carefully, from small to bigger.
As for C60 way of action, I speculate that it neutralizes the poisonous aldehydes (MDA and 4-HNE) which are product of the normal metabolism and/or cleaves protein cross-links caused by these aldehydes (improving skin collagen). I am doing some research in chemistry now.

Live longer and prosper, man.

[nickdino]

hi, i came across this which if correct is a bit of an eye opener perhaps.

In 2006, a JAMA study concluded that, "patients and physicians should remain cautious about extrapolating the findings of prominent animal research to the care of human disease," and that "even high-quality animal studies will replicate poorly in human clinical research."
What’s more, mice are used in nearly 60% of all experiments — and perhaps even more so in life extension research. As Slate's Daniel Engber has argued, mice are among the most unreliable test subjects when it comes to approximating human biological processes. "It's not at all clear that the rise of the mouse — and the million research papers that resulted from it — has produced a revolution in public health," he has said.

He points to research in caloric restriction, for example, a dietary regimen that (arguably) confers life extending benefits. The present consensus is that caloric restriction extends life in mice, but not very much in primates.
Moreover, according to Estep, food fed to mice in the labs is basically junk food — about 70% of calories from starch and sugar. “I sometimes call typical caloric restriction experiments in mice "Cookie Restriction," he says. “It’s not surprising that some mice live longer if fed less of this stuff.”

http://io9.com/do-th...an-co-486041314

[Mind]

Thanks for the reminder nickdino....it is something I post quite often as well.

Members here are well aware of the risk/reward and the basic science surrounding C60. They are willing to experiment in order to benefit humanity (the LongeCity crowd is pretty awesome that way)

[pleb]

Hi Mind I think your idea of getting rat fans and rat clubs interested is a damned good one, they are much more likely to see this through to the end than someone who doesn't normally keep them as pets, for none rat fans who volunteer enthusiasm can be good at the start but wane after a year or two,

[Mind]

I contacted one large group last year, but it was a difficult sell. Because they were mostly non-scientists and did not seem too have a lot of science aptitude there was a general feeling of distrust/fear, even though I tried my darndest to explain it simply and highlight the potential benefits. Maybe if AgeVivo's mice make it a few more months (at least) then more people will be willing.

[VP.]

Maybe what we need is a rat/mouse study to see to if there is any truth to the endurance improvements reported by many users. A treadmill study after C60 dosing would be relatively inexpensive and the results could be reported in months rather then years. It would certainly get athletes attention. Here is a rat treadmill for sale: http://www.accuscan-...-Treadmill/1902

[niner]

Sooner or later, I think that the secret is going to get out into the endurance sports community, with or without animal studies. Sarah Vaughter is running a story about Boris Sala, 43, who has competed in the Dutch national indoor rowing championships for a number of years with consistent, though not winning performance. Then he got onto a c60-oo protocol, and now he's the national champion. This is a no-brainer for cyclists.

[AgeVivo]

idea of getting rat fans and rat clubs interested

one year ago Logic and I found online a list of rats & mouse rat fan clubs all over the world. We've sent emails to all of them, explaining what it is in large, and got virtually no answer: only a few "I might be interested" but with no late answer. No negative answer. We don't know what the reason is but it might be that those persons
1) are busy
2) do not feel strong enough to be followed by scientists, and do not speak English well
3) that when asked their veterinarians advise them that it is probably hoax

That's what I perceived through my contact: I got contacts Crevetterbelle through a rat fan friend who is too busy (1) to do the experiment but gave me contacts, Both my friend and Crevertterbelle expressed (1) and (2). My friend's veterinarian was quite skeptical (3) thought he asked for papers, she transmitted the request to me, I sent papers.. and he thought that such a placebo controlled test can bring something. Crevetterbelle caught very well the scientific interest but, as we have seen, she is very busy and it is not easy for her to set the experiment.

So as we can see, yes rat fans are great but there is a strong barrier and the best way is to find rat fans through one's group of friends and be ready to explain the thing (http://www.longecity...lates/c60health).

Edited by AgeVivo, 08 May 2013 - 01:58 PM.

[Turnbuckle]

idea of getting rat fans and rat clubs interested

in fact Logic and I found online a list of rats & mouse rat fan clubs all over the world. We've sent emails to all of them, explaining what it is in large, and got virtually no answer: only a few "I might be interested" but with no late answer. No negative answer. We don't know what the reason is but it might be that those persons
1) are busy
2) do not feel strong enough to be followed by scientists, and do not speak English well
3) that when asked their veterinarians advise them that it is probably hoax

Most likely it's because you don't have the credentials to be doing this. And if you had them, you would be running your own tests on your own rats. Also, people who like rats and mice generally don't like scientists experimenting on their animals.

[pleb]

i've talked to logic about this and i agree with him that the controls in this situation are not really needed there are thousands of published papers showing the average life span of rats is between 2 and 3 years, unless this is for eventual publication in a journal ,
having a seperate group of control that last as long as the millions of other rats that have been used over the years won't make the results any less,viable,that's just my opinion

[AgeVivo]

Most likely it's because you don't have the credentials to be doing this

In the letter I stated that I am a former researcher at the Pasteur Institute, did mouse lifespan tests, I am in contact with the authors of the paper, and have starting with a few mice at home. The answers I've had were quite respectful in that regards, so... I don't know. It is true that otherwise it was an email coming from nowhere. That's why I think that the key thing is to use our existing contacts, otherwise it may sound like spam.

[AgeVivo]

i've talked to logic about this and i agree with him that the controls in this situation are not really needed there are thousands of published papers showing the average life span of rats is between 2 and 3 years, unless this is for eventual publication in a journal ,
having a seperate group of control that last as long as the millions of other rats that have been used over the years won't make the results any less,viable,that's just my opinion

yes if they live 3.5+ year old, otherwise to a certain degree: there are various types of strains with some variation of lifespans, and the animals may be better cared than as usual. If it is treated mice that live 2.5 or 3 years on average, controls would help judge. Also there are some claims of mice and rats having lived very long without treatment, one of which won the Mprize; I look at those cases with suspicion (if someone tells you he is 150 year old...) but not everyone in the field.

Also, indeed, there is a matter of "demonstrating" things (eg a p_value) for the person who refuses to bother with such vague considerations. Eg for a paper but also to convince "fast-interpreting"-scientists.