131 replies to this topic

[bixbyte]

 

http://www.jimmunol....gAbstracts/55.2

 

kmoody, this is exiting work, thanks for keeping us informed on your progress.

 

I was drawn to this forum because of a thoughtful post by niner on fullerenes and asthma.  In regards to the study design, the study linked above (for some reason I can't link inline) had two treatment groups, one treated during the development of asthma and another treated after.  So they were able to test both the preventative and "after infection" treatment value of C70 for asthma.

 

On a different topic, could someone help put these initial results in layman terms?  Is this indicating that C60oo treatment is causing more tumors to form in bone marrow?

The C60oo treated mice have double the tumor burden within the bone marrow as compared to controls. Most of our C60oo treated mice also have palpable tumors whereas most controls do not. We have a few working hypotheses to explain this data.

 

It could be that C60oo exacerbates cancer. However, I do not believe this to be the case. Our pilot study showed a very compelling, dose-dependent decrease in all-cause mortality.

 

I am aware of three major changes to the study design between the pilot and our current study. One was a switch from CIEA NOG to NOD/SCID mice. These are very similar immunocompromised mouse models that I don't think are sufficiently distinct to cause this discrepancy, but an essential non-obvious difference could exist. Another possibility is the lag time with treatment. In the pilot study we began treating with C60oo when the mice were inoculated with the leukemia cells. In the repeat, we wanted to clearly differentiate the effects of prophylactic treatment vs. reduced engraftment of cancer vs. cancer treatment. We chose to study the cancer treatment, and waited until 1 week after the cancer was injected before beginning treatment. It could be that properties of C60 reduce tumor engraftment but exacerbate cancer. Again, I do not believe this to be the case.

 

Our current lead hypothesis is sourcing. For our pilot study, we made C60oo fresh in-house. At the time of the current study, we wanted to add quality assurance to the study design so we chose to go with a vendor that claimed robust product specifications. We have 3 ongoing studies that are using C60oo sourced from that vendor, including the leukemia one described here. In all three studies, the C60oo treated mice are trending heavily in the direction of accelerated death from all-cause mortality. In our repeat lifespan study, we have confirmed by blinded 3rd party histopathology that at least some C60oo treated mice are dying of cancer. The product from this vendor has failed every one of our quality control tests as we have brought them online in-house. We have repeatedly asked for CoA and product specification documents but these forms have not been provided.

 

So in layman terms, something bad appears to be happening to these mice. A plausible common denominator is that C60oo sourcing is incredibly important. Some vendors may have a product that could promote all the health benefits we hear about with C60oo. Some vendors may have a product that can dramatically increase all cause mortality. We are aggressively studying the differences between our formulation and that of the vendor to understand what the difference may be.

 

 

It's valid to defend the aim of an experiment , I think what's being said here is that there could be a different aim.

 

Not taking anything away from Ichor, however:

 

there is little monetization available for a preventative/prophylactic anything, and I don't believe that the development of prophylactic drugs is part of Ichor's business model -- except as perhaps part of the contract research.

 

With the emerging data that shows environmental and lifestyle choices are overwhelmingly the cause of cancer, a cancer preventative would face a huge epidemiological and evidence battle, as would any "life-extension" drug.

 

Marie Calment ate 2 lbs of dark chocolate a week and reportedly drowned her food (most meals) in olive oil -- combination of anti-oxidant and olive oil  -- correlated? causal? how long a study would suffice.

 

Tweak a C60OO molecule enough to get a patent, and show efficacy against existing disease and you have a cheap to manufacture almost priceless drug.

 

Truvada was initially a  HAART treatment for AIDS and HIV patients, that just so happens to be an extremely potent and efficacious pre-exposure prophylactic anti-viral with respect to HIV.

 

I agree with sensei on this one. Happy to run contract research near cost for the benefit of the community, but the value to us as a business is in drugs. That said, there is obvious overlap in the safety profile of C60oo as a drug candidate or a prophylactic supplement, so robust characterization of manufacturing, formulation, stability, and safety issues is where everyone's interests are likely aligned. Note that the initial study was not really meant to show C60oo can treat cancer. Rather, it was to see if it would exacerbate a human cancer model in mice.

 

" We chose to study the cancer treatment, and waited until 1 week after the cancer was injected before beginning treatment. "

 

And you blame the results on the C60 Olive Oil ?

I do not think you understand how cancer proliferates.

Cancer injected turns into every kind of cancer.

 

So, I still suggest the same procedure as I did in your original pilot study posted results and your second study:

 

First treat these test animals with C60 Olive Oil for a week or more.

And then inoculate them with the Human Cancer cells. 

Do the drug trial similar to Baati.

That is what everyone wants to know.

[stefan_001]

http://www.jimmunol....gAbstracts/55.2
 
kmoody, this is exiting work, thanks for keeping us informed on your progress.
 
I was drawn to this forum because of a thoughtful post by niner on fullerenes and asthma.  In regards to the study design, the study linked above (for some reason I can't link inline) had two treatment groups, one treated during the development of asthma and another treated after.  So they were able to test both the preventative and "after infection" treatment value of C70 for asthma.
 
On a different topic, could someone help put these initial results in layman terms?  Is this indicating that C60oo treatment is causing more tumors to form in bone marrow?

The C60oo treated mice have double the tumor burden within the bone marrow as compared to controls. Most of our C60oo treated mice also have palpable tumors whereas most controls do not. We have a few working hypotheses to explain this data.
 
It could be that C60oo exacerbates cancer. However, I do not believe this to be the case. Our pilot study showed a very compelling, dose-dependent decrease in all-cause mortality.
 
I am aware of three major changes to the study design between the pilot and our current study. One was a switch from CIEA NOG to NOD/SCID mice. These are very similar immunocompromised mouse models that I don't think are sufficiently distinct to cause this discrepancy, but an essential non-obvious difference could exist. Another possibility is the lag time with treatment. In the pilot study we began treating with C60oo when the mice were inoculated with the leukemia cells. In the repeat, we wanted to clearly differentiate the effects of prophylactic treatment vs. reduced engraftment of cancer vs. cancer treatment. We chose to study the cancer treatment, and waited until 1 week after the cancer was injected before beginning treatment. It could be that properties of C60 reduce tumor engraftment but exacerbate cancer. Again, I do not believe this to be the case.
 
Our current lead hypothesis is sourcing. For our pilot study, we made C60oo fresh in-house. At the time of the current study, we wanted to add quality assurance to the study design so we chose to go with a vendor that claimed robust product specifications. We have 3 ongoing studies that are using C60oo sourced from that vendor, including the leukemia one described here. In all three studies, the C60oo treated mice are trending heavily in the direction of accelerated death from all-cause mortality. In our repeat lifespan study, we have confirmed by blinded 3rd party histopathology that at least some C60oo treated mice are dying of cancer. The product from this vendor has failed every one of our quality control tests as we have brought them online in-house. We have repeatedly asked for CoA and product specification documents but these forms have not been provided.
 
So in layman terms, something bad appears to be happening to these mice. A plausible common denominator is that C60oo sourcing is incredibly important. Some vendors may have a product that could promote all the health benefits we hear about with C60oo. Some vendors may have a product that can dramatically increase all cause mortality. We are aggressively studying the differences between our formulation and that of the vendor to understand what the difference may be.
 

It's valid to defend the aim of an experiment , I think what's being said here is that there could be a different aim.

 
Not taking anything away from Ichor, however:
 
there is little monetization available for a preventative/prophylactic anything, and I don't believe that the development of prophylactic drugs is part of Ichor's business model -- except as perhaps part of the contract research.
 
With the emerging data that shows environmental and lifestyle choices are overwhelmingly the cause of cancer, a cancer preventative would face a huge epidemiological and evidence battle, as would any "life-extension" drug.
 
Marie Calment ate 2 lbs of dark chocolate a week and reportedly drowned her food (most meals) in olive oil -- combination of anti-oxidant and olive oil  -- correlated? causal? how long a study would suffice.
 
Tweak a C60OO molecule enough to get a patent, and show efficacy against existing disease and you have a cheap to manufacture almost priceless drug.
 
Truvada was initially a  HAART treatment for AIDS and HIV patients, that just so happens to be an extremely potent and efficacious pre-exposure prophylactic anti-viral with respect to HIV.

I agree with sensei on this one. Happy to run contract research near cost for the benefit of the community, but the value to us as a business is in drugs. That said, there is obvious overlap in the safety profile of C60oo as a drug candidate or a prophylactic supplement, so robust characterization of manufacturing, formulation, stability, and safety issues is where everyone's interests are likely aligned. Note that the initial study was not really meant to show C60oo can treat cancer. Rather, it was to see if it would exacerbate a human cancer model in mice.

" We chose to study the cancer treatment, and waited until 1 week after the cancer was injected before beginning treatment. "
 
And you blame the results on the C60 Olive Oil ?
I do not think you understand how cancer proliferates.
Cancer injected turns into every kind of cancer.
 
So, I still suggest the same procedure as I did in your original pilot study posted results and your second study:
 
First treat these test animals with C60 Olive Oil for a week or more.
And then inoculate them with the Human Cancer cells. 
Do the drug trial similar to Baati.
That is what everyone wants to know.

I think it's important to know both. There may be people starting to use it who are not cancer free.

[kmoody]

Once again thanks very much for your dilligence to the c60 community; is there are anyway of helping fund the cost of your research?

 

 

I wonder if it is useful to study a group of mice on a course of your in-house c60 after being previously being dosed with the apparent dodgy batch. It might be useful information for those concerned about the product they've taken.

Thanks for the offer to help with funding, but lets hold off on that discussion for now. We have some questions at hand that need to be addressed. Once answered, they may or may not support future research directions. We're comfortably funded with the grant from Methuselah Foundation to get these things figured out.

 

It could be useful to give good C60oo after this apparent bad batch, but that would significantly confound any results we obtain from ongoing studies. If we finish them as planned and see an increase in all cause mortality, then redo them with fresh C60oo and see a decrease in all cause mortality, we will have data strongly supporting that the vendor's C60oo is problematic. I don't think we can assume any outcomes despite clear trends in all of my mice. Unfortunately that just isn't good science. We have to answer one question at a time. No cutting corners.

 

" We chose to study the cancer treatment, and waited until 1 week after the cancer was injected before beginning treatment. "

 

 

 

And you blame the results on the C60 Olive Oil ?

I do not think you understand how cancer proliferates.

Cancer injected turns into every kind of cancer.

 

So, I still suggest the same procedure as I did in your original pilot study posted results and your second study:

 

First treat these test animals with C60 Olive Oil for a week or more.

And then inoculate them with the Human Cancer cells. 

Do the drug trial similar to Baati.

That is what everyone wants to know.

The C60oo issues are not confined to the leukemia study, which suggests that the C60 olive oil is to blame, not necessarily the cancer. If the positive effects of C60oo were indeed prophylactic, I would expect C60oo treated mice to have similar outcomes as the control mice, not accelerated death. This trend is consistent among three ongoing studies, 1) leukemia, 2) radiation syndrome, and 3) lifespan. I am unclear what you mean by "cancer injected turns into all kinds of cancer". That is not accurate. The kg1a cells used are a human myeloid leukemia line that remains confined within the myeloid lineage, and predominantly proliferates as a myeloid progenitor cell type.

 

There are two different questions that can be asked, 1) does C60 prevent cancer, and 2) does C60 treat cancer. These must be separated to get clear results, particularly in the context of this cancer model. If we see efficacy in a treatment C60oo group, that will provide us with critical data necessary to obtain additional funding to further study C60oo in different models. I do not see a need to run a prophylactic arm of the leukemia study because we are running a C60oo arm of an ongoing lifespan study, which should tell us something interesting about C60oo's ability to prevent cancer. However, I suspect that will need to be restarted because we used this vendor's C60oo, and are seeing a strong trend of C60oo treated mice dying as compared to controls. We have C60oo treated mice in the lifespan study that have died from cancer, suggesting either C60oo is not a magic bullet for preventing cancer, or perhaps that the C60oo we received is no good (our working hypothesis).

 

Baati did not do a drug trial. He looked at a model of induced liver toxicity. I'm not sure what you mean by this. Are you able to clarify? We are doing something similar with a radiation syndrome study.

 

I think it's important to know both. There may be people starting to use it who are not cancer free.

I agree. Again, funding is an ever present issue. If someone wants to cut me a check for a few hundred thousand I would be happy to look at whatever they want. Else, I need to ensure the studies we decide to do are appropriate to leverage further funding if interesting results are observed.

 

Of note, our company may be expanding our vivarium in the near future as we are currently running at capacity. It is possible that we could take on studies to answer some of these additional questions in the future. So please, do keep posting questions. Even if we can't answer them now the questions could be very helpful to guide studies in the future.

Edited by kmoody, 25 December 2015 - 08:58 PM.

[sensei]

 

 

The C60oo issues are not confined to the leukemia study, which suggests that the C60 olive oil is to blame, not necessarily the cancer. If the positive effects of C60oo were indeed prophylactic, I would expect C60oo treated mice to have similar outcomes as the control mice, not accelerated death. This trend is consistent among three ongoing studies, 1) leukemia, 2) radiation syndrome, and 3) lifespan. 

 

 

That is extremely interesting as I and others have seen notable increases in the amount of ionizing radiation necessary to induce acute radiation sickness in epithelial cells (human sunburn); as well as shortened duration of healing.

 

Unless, in your trial, the C60OO is administered post radiation challenge.

 

Many many experiences point to C60 as being preventative.

 

Even the effect on alcohol intoxication has never worked for me if I take the C60 post drinking -- in fact, during one experiment, consumption of approx 30 mg of C60 after consuming a bottle of wine plus a few glasses I experienced one of the worst hangovers in my life later that evening and throughout the next day .

 

I also believe the life extension mechanism seen in Baati is due to the large acute dosage protocol causing some sort of effect. That is the only reasonable assumption I can make as there is no long term low dose protocol results to compare to.  This is one of the reasons why my doses are usually 30 mg+

 

That is not to say that there are not benefits reported across the community from long term low dose protocol, nor that the same effect can be achieved from long term low dose protocols.

 

However, High Desert Wizard and I have ONLY seen the dramatic changes after commencing high dosage protocols whether acute, long term or both.

[smithx]

Yes, SES does (or at least did) use sonication rather than following Baati's methodology. See my thread here:

http://www.longecity...e-oil-supplier/

 

I called them up at that time and asked them about their process in detail, and what I wrote is what they told me on the phone.

 

 

[bixbyte]

There are two different questions that can be asked, 1) does C60 prevent cancer, and 2) does C60 treat cancer. These must be separated to get clear results, particularly in the context of this cancer model. If we see efficacy in a treatment C60oo group, that will provide us with critical data necessary to obtain additional funding to further study C60oo in different models. I do not see a need to run a prophylactic arm of the leukemia study because we are running a C60oo arm of an ongoing lifespan study, which should tell us something interesting about C60oo's ability to prevent cancer. 

 

 

 

Here is what I see in your first test your results for the mice with the largest dose of C60 showed good results and in your second test your results show failure that you blame on the C60 vendor.

So, in order for you to have success in your business venture you need to repeat the original results.

 

Second since you see no need to do an arm prophylactic that might result in no need for your medication to come to market.

 

That is what I see.  

 

I have read numerous cancer drug studies for many years.

Specially as a long term investor in startups and secondly to gain insight into the science.    

 

 

 

[ambivalent]

It might be a long shot but I wonder if Moussa still has any of the oil left from his experiment. I seem to recall he supplied Agevivo with some that was left from his study, which I believe would have been quite old at the time, and all of Agevivo's mice had tumours at the time of death. If the oil was available it would be interesting to know if it now would fail the quality testing in the same way as the SES mix.

Edited by ambivalent, 26 December 2015 - 12:52 PM.

[Wilberforce]

Agree with Sensei's comment on bad hangover when taking c60 after alcohol.

[Invariant]

kmoody, and others: would it perhaps be a good idea to involve prof. Fathi Moussa or another expert on the chemistry of C60 in this discussion? Maybe they can guess what the problem with the new batch of C60 is, and tell us whether sonication is indeed a bad idea.

[kmoody]

 

Here is what I see in your first test your results for the mice with the largest dose of C60 showed good results and in your second test your results show failure that you blame on the C60 vendor.

So, in order for you to have success in your business venture you need to repeat the original results.

 

Second since you see no need to do an arm prophylactic that might result in no need for your medication to come to market.

 

That is what I see.  

 

I have read numerous cancer drug studies for many years.

Specially as a long term investor in startups and secondly to gain insight into the science.    

 

We already have a C60oo arm in an ongoing lifespan study. These mice are also dying faster than controls. We were curious as to why, so we began having blinded 3rd party histopathology on necropsy. In the C60oo treated mouse that was tested, cause of death was cancer. We will be analyzing others as they die. I understand that you think prophylactic treatment is worth investigating. I want to clearly articulate that we are already assessing that in our ongoing lifespan study. However, the data we have so far shows a clear trend of C60oo treated mice dying prematurely and at least the one we've tested so far (we will be testing others) died of cancer.

 

The xenograft model cannot accurately assess prophylactic efficacy because the cancer cells are not generated de novo, they are grown ex vivo and engrafted. The lifespan study is answering the question about prophylactic treatment. The xenograft model is answering the question about therapeutic effects. These are two different questions that require different models to answer them reliably.

 

The consistent trend that is supported by all available data is that the C60 supplied by the vendor is responsible. This is a hypothesis that is supported by all quality assurance testing we have performed since noticing the trend, and one which we are continuing to aggressively test.

 

Success in my business has nothing to do with seeing results we want to see in this study. We are basically running these studies at cost for the good of the community, and doing lots of pro bono work besides (for example, quality control of C60oo was NOT in the job description of any project I have received funding for, we are doing this because we want to do good science). If I wanted to "prove" C60oo is a good cancer drug I would simply not report the data we are currently seeing and instead wait until the study is repeated and shows what I want it to show. My interest here is providing a public service announcement to members of the Longecity community who may be consuming C60oo from SES research. In my laboratory, we are seeing a clear trend in increased all-cause mortality in three different ongoing studies that seems to be attributable to this product. I thought some folks here might be interested in knowing that.

 

So to be clear -- we ARE assessing the prophylactic effects of C60oo in mice via the lifespan study and available data suggests that those mice ARE DYING FASTER than controls. We are ALSO assessing the direct therapeutic benefits of C60oo in mice that already have cancer. In ALL CASES C60oo appears to be bad. We are not simply blaming this on the vendor. Quality issues with the product is our working hypothesis and this position is based on all available data. We are continuing to work to confirm this.

 

kmoody, and others: would it perhaps be a good idea to involve prof. Fathi Moussa or another expert on the chemistry of C60 in this discussion? Maybe they can guess what the problem with the new batch of C60 is, and tell us whether sonication is indeed a bad idea.

We could, though my biochemist and organic chemist are both already working on this with me. We're moving as fast as we can towards finding answers, but things are a bit slow on account of the holidays and that sonication is just one possible explanation of several that we are investigating.

Edited by kmoody, 26 December 2015 - 05:35 PM.

[sensei]

 

 

So to be clear -- we ARE assessing the prophylactic effects of C60oo in mice via the lifespan study and available data suggests that those mice ARE DYING FASTER than controls. 

 

 

 

Was the protocol the same as Baati?

 

1.7 mg/kg by gavage daily for a week, then other dosing every two weeks(or scaled for mice lifespan vs rat) for a total of 24 doses @ 1.7 mg/kg?

 

Were the mice age-matched to the mice in the Baati study, wrt time of initial treatment?

Edited by sensei, 26 December 2015 - 06:25 PM.

[bixbyte]

 

 

 

Just another question:

Can your chart be right?

You have detected C70 signature in your test?

Maybe someone needs to contact sesres?

 

 

 

Purchase C60 in Olive Oil

As the nations largest manufacturer and supplier of scientific grade Carbon 60, we've produced a 100ML C60 Olive Oil product. The purest form of C60 was used as well as certified organic extra virgin olive oil.

At SES Research, we understand this research has not been tested on humans and don't recommend human consumption. We manufacturer and stock Carbon 60 (C60) in its pure state for sale by the gram.

 

 

https://sesres.com/C60-olive-oil.asp

 

 

 

[sensei]

 

 

 

Just another question:

Can your chart be right?

You have detected C70 signature in your test?

Maybe someone needs to contact sesres?

 

 

 

Purchase C60 in Olive Oil

As the nations largest manufacturer and supplier of scientific grade Carbon 60, we've produced a 100ML C60 Olive Oil product. The purest form of C60 was used as well as certified organic extra virgin olive oil.

At SES Research, we understand this research has not been tested on humans and don't recommend human consumption. We manufacturer and stock Carbon 60 (C60) in its pure state for sale by the gram.

 

 

https://sesres.com/C60-olive-oil.asp

 

C70 is known as the primary (overwhelming) contaminant in C60 -- even at 99.95% there is still C70 contamination 

 

The chart is correct

[pampoenkop]

The C70 was added to the samples at a known concentration so that they could account for any losses during the extraction process. This was mentioned in the first post and later explained in more detail a few post down.

[sensei]

The C70 was added to the samples at a known concentration so that they could account for any losses during the extraction process. This was mentioned in the first post and later explained in more detail a few post down.

 

 

Still does not change the FACT that the primary contaminant of C60 is C70.

[bixbyte]

My interest here is providing a public service announcement to members of the Longecity community who may be consuming C60oo from SES research. In my laboratory, we are seeing a clear trend in increased all-cause mortality in three different ongoing studies that seems to be attributable to this product. I thought some folks here might be interested in knowing that.

 

So to be clear -- we ARE assessing the prophylactic effects of C60oo in mice via the lifespan study and available data suggests that those mice ARE DYING FASTER than controls. We are ALSO assessing the direct therapeutic benefits of C60oo in mice that already have cancer. In ALL CASES C60oo appears to be bad. We are not simply blaming this on the vendor. Quality issues with the product is our working hypothesis and this position is based on all available data. We are continuing to work to confirm this.

 

 

 I do not see a need to run a prophylactic arm of the leukemia study because we are running a C60oo arm of an ongoing lifespan study, which should tell us something interesting about C60oo's ability to prevent cancer. However, I suspect that will need to be restarted because we used this vendor's C60oo, and are seeing a strong trend of C60oo treated mice dying as compared to controls. We have C60oo treated mice in the lifespan study that have died from cancer, suggesting either C60oo is not a magic bullet for preventing cancer, or perhaps that the C60oo we received is no good (our working hypothesis).

 

 

 

First of all, you sound like you are contradicting yourself?

 

Second, there are members on this board that have been dosing on C60 Olive Oil for many years. 

Any of the members dosing with C60 olive oil now have cancer?

(That is a question maybe could be answered by the c60 poll?)

 

Assessing the prophylactic effect without actually preforming a prophylactic arm?? 

 

I certainly do not see how you can determine that the members that have been dosing with C60 and are NOT 

injected with Leukemia could have a similar life expectancy to your Leukemia injected mice?

 

Are you informing the experimenters that supplement to cease consuming C60 Olive Olive?

 

I am sure lucky I read this board for 2 years before I tested one #0 cap of C60 olive oil / day.

I did not want to be a guinea pig!

 

 

 

[aconita]

Sonification involves a huge heat generation within the micro bubbles which may in some way alter the olive oil, the temperature is by far above the 200 c of the olive oil smoking point even if for only a very short amount of time...but still some changes are likely to occur.

 

Not supported by any scientific evidence but intuition leads me to believe it is likely that "amplifying" is one of the C60 mechanism of action, we all know that olive oil itself is healthy, promotes longer lifespan and has anti carcinogenic proprieties therefore if my feeling is right adding C60 amplifies those health effects of olive oil.

 

We also know heating above a certain point oils is going to turn them very unhealthy, if C60 amplifies those unhealthy effects too maybe we have got kind of an explanation for sonificated C60oo showing disturbing and contradictory results.

 

It seems in facts that above smoking point oils tends to turn carcinogenic.

 

I know, it is a bit out of topic and not very scientific but...  

[kmoody]

 

 

 

So to be clear -- we ARE assessing the prophylactic effects of C60oo in mice via the lifespan study and available data suggests that those mice ARE DYING FASTER than controls. 

 

 

 

Was the protocol the same as Baati?

 

1.7 mg/kg by gavage daily for a week, then other dosing every two weeks(or scaled for mice lifespan vs rat) for a total of 24 doses @ 1.7 mg/kg?

 

Were the mice age-matched to the mice in the Baati study, wrt time of initial treatment?

 

We are administering approximately 4mg/kg in pre-aged C57BL/6 mice by spiking the diet. Mouse weights and food consumption are being monitored so the dosing won't be perfect but we can take into account during final data analysis. We discussed the pros and cons with gavage feeding with the sponsor but it was determined that for cost and practicality using a diet-based administration made the most sense. Note that we are not using a few rats like Baati. There are 45 mice per group so the study is well powered. Mice were all age and gender matched.

 

The C70 was added to the samples at a known concentration so that they could account for any losses during the extraction process. This was mentioned in the first post and later explained in more detail a few post down.

Pampoenkop is correct. This graph just shows we can easily resolve C60 and C70. When we extract C60 from tissues or organs we spike the sample with a known concentration of C70. This allows us to adjust the final concentration of C60 measured to reflect loss during the extraction process. So in response to bixbyte's question yes, our results are accurate. But no, I am not claiming there is significant contamination of C70 in the C60. This graph simply shows we can resolve the two from one another, this graph is not a QC graph on C60 or anything like that. Just a quick snapshot during method development.

 

 

First of all, you sound like you are contradicting yourself?

 

Second, there are members on this board that have been dosing on C60 Olive Oil for many years. 

Any of the members dosing with C60 olive oil now have cancer?

(That is a question maybe could be answered by the c60 poll?)

 

Assessing the prophylactic effect without actually preforming a prophylactic arm?? 

 

I certainly do not see how you can determine that the members that have been dosing with C60 and are NOT 

injected with Leukemia could have a similar life expectancy to your Leukemia injected mice?

 

Are you informing the experimenters that supplement to cease consuming C60 Olive Olive?

 

I am sure lucky I read this board for 2 years before I tested one #0 cap of C60 olive oil / day.

I did not want to be a guinea pig!

 

I'm not sure how clearer I can be on this, but let me try...

 

The cancer study we are running is called a xenograft study. What this means is that we have an immunocompromised mouse strain called NOD/SCID that is capable of receiving tissues from a different species. NOD/SCID mice do not reject these tissues because they lack an adaptive immune system, which is normally responsible for detecting, flagging, and eliminating foreign tissues.

 

For our xenograft study, we are injected NOD/SCID mice with the human acute myeloid leukemia cell line kg1a. Xenograft models differ from de novo cancer formation because the cancers are already fully formed when the mice receive them. In contrast, de novo cancer formation involves the progressive accumulation of some dozen or so mutations before precancerous lesions become cancerous.

 

The utility of xenograft studies is highly dependent upon engraftment efficiencies. Some cancers have very high success rates of engraftment when given to NOD/SCID mice (including this model), whereas others are low. We have some cancer lines that do not engraft at all, whereas we have others that are at or near 100%. Because of this, engraftment efficiencies are a large confounding variable in any xenograft model.

 

Suppose we wanted to design a prophylactic study (as you suggest) with a treated and untreated arm to see if there is an increase in lifespan. This does not really tell us what is going on. In such a model, we cannot say the prophylactic treatment does anything to prevent cancer formation because the cancer was already formed when the mouse received it. If we see efficacy, it may simply be that the treatment is decreasing engraftment efficiency (recall this is an ARTIFACT of the model, not something that happens in any appreciable way in real life). The other option to explain efficacy is that there is cancer killing or growth inhibition. So in a prophylactic arm, positive outcomes could mean that the study was compromised because of the engraftment artifact, or it could be that there is therapeutic benefit. The simple way to distinguish which is to start the treatment AFTER engrarftment has occurred, so any positive effect seen can be attributable directly to therapeutic benefit. This is what we did in the present study.

 

There are appropriate models to assess prophylactic effects of C60oo. For example, you could just give a bunch of mice C60oo and track them for a long time to see if there is a difference in rates of cancer formation. This is precisely what we are doing in the lifespan model. Alternatively, you could prophylactically treat a group of mice with C60oo then induce cancer formation by administering carcinogens. These sorts of models have their own artifacts and confounding variables, depending on the study design and model selected.

 

Because of these reasons, we cannot assess the prophylactic effects of C60oo with a prophylactic arm of the xenograft study. This could be tested within the scope of our ongoing lifespan study, which we are doing. However, the mice in the lifespan study that have been treated with C60oo are showing a strong trend of premature death from all-cause mortality.

 

I am aware that there are many members on this board that have been dosing C60oo for many years. I am not suggesting that C60oo treatment causes cancer. I am simply stating that treatment with a C60oo product of one particular vendor is showing a strong trend of increased cancer burden in our xenograft model, and is also showing a strong trend of increased all-cause mortality in three ongoing studies. I make no further claims than that. I make no claims about members that are self dosing with C60oo. There are too many confounding variables that could explain any differences. For example, many that are dosing are doing so with small doses. Many are using home-made product or product from other vendors. So far as I am aware, none of the people on this forum are mice so inter-species differences cannot be excluded. Finally, there is selection bias. I suspect we haven't heard much from anyone who takes C60oo that has died of cancer.

 

I make no explicit claims to people who are self experimenting. I am simply reporting the data from my lab and providing my opinion about what could be happening. I'll leave extrapolations and generalizations to other people because that is outside of my job description.

Edited by kmoody, 28 December 2015 - 05:33 PM.

[Major Legend]

Dosing many years does not mean the substance is not carcinogenic in the long term. The original study was pubished in 2012, thats only 3 years. To think I dosed tablespoons of the stuff at some point - yikes.

 

I suppose most people are reporting good effects. I am just hoping that our method did not produce the same C60OO you are using from SES.

 

At the very least your studies are proving (if your hypothesis is right) that we were right about the fact that C60 could easily be in other forms and the quality of it can vary. The forum format is good and bad at the same time, the possibility that C60 purity could easily be disrupted was discussed very early on in the C60 thread.

 

I think the sonification is potentially where it went wrong - the major difference between the original study was that I think he didn't actually

grind the C60 or break it down in any vigorous manner. It's reasonable to assume smashing C60 down exposes it to far more air than normal, and may be increasing the amount of c60 deriviatives.

 

As I understand C60 can go both ways, it can also turn into a super oxidant. I think we should be more cautious going forward, afterall this is nanotechnology and technically this isn't so much a drug, but a molecule that can go do things that no current drugs can do, which places it in it's own class of both possibility and danger.

 

Edited by Major Legend, 28 December 2015 - 09:40 PM.

[Turnbuckle]

 It's reasonable to assume smashing C60 down exposes it to far more air than normal, and may be increasing the amount of c60 deriviatives.

 

C60 will neither react with O2 in the atmosphere nor break down in a mortar. So nothing to worry about. Ultrasonic treatment in oil, however, will create a spectrum of oil radicals that will then react with the C60, producing products that are quite different from those used in the original paper.

 

 

Degradation of edible oil during food processing by ultrasound: electron paramagnetic resonance, physicochemical, and sensory appreciation.

 

Abstract

During ultrasound processing of lipid-containing food, some off-flavors can be detected, which can incite depreciation by consumers. The impacts of ultrasound treatment on sunflower oil using two different ultrasound horns (titanium and pyrex) were evaluated. An electron paramagnetic resonance study was performed to identify and quantify the formed radicals, along with the assessment of classical physicochemical parameters such as peroxide value, acid value, anisidine value, conjugated dienes, polar compounds, water content, polymer quantification, fatty acid composition, and volatiles profile. The study shows an increase of formed radicals in sonicated oils, as well as the modification of physicochemical parameters evidencing an oxidation of treated oils.

 

http://www.ncbi.nlm....pubmed/22804736

 

 

 

Edited by Turnbuckle, 28 December 2015 - 09:52 PM.

[Major Legend]

 

 It's reasonable to assume smashing C60 down exposes it to far more air than normal, and may be increasing the amount of c60 deriviatives.

 

C60 will neither react with O2 in the atmosphere nor break down in a mortar. So nothing to worry about. Ultrasonic treatment in oil, however, will create a spectrum of oil radicals that will then react with the C60, producing products that are quite different from those used in the original paper.

 

 

http://blogs.lt.vt.e...in-ambient-air/ - it does react with air according to this, air does seem a little benign as a cause - I was just trying to point out that C60 does react and turn into other things.

 

edit: oh maybe there is no Ozone in normal air, sorry for the messup.

 

But yes - I think sonification may be the issue there in regards to the oil radicals, from what I've read C60 can become a huge number of different things.

Edited by Major Legend, 28 December 2015 - 10:00 PM.

[sensei]

 

 

 

 

So to be clear -- we ARE assessing the prophylactic effects of C60oo in mice via the lifespan study and available data suggests that those mice ARE DYING FASTER than controls. 

 

 

 

Was the protocol the same as Baati?

 

1.7 mg/kg by gavage daily for a week, then other dosing every two weeks(or scaled for mice lifespan vs rat) for a total of 24 doses @ 1.7 mg/kg?

 

Were the mice age-matched to the mice in the Baati study, wrt time of initial treatment?

 

We are administering approximately 4mg/kg in pre-aged C57BL/6 mice by spiking the diet. Mouse weights and food consumption are being monitored so the dosing won't be perfect but we can take into account during final data analysis. We discussed the pros and cons with gavage feeding with the sponsor but it was determined that for cost and practicality using a diet-based administration made the most sense. Note that we are not using a few rats like Baati. There are 45 mice per group so the study is well powered. Mice were all age and gender matched.

 

 

 

 

Can you be a bit more clear?

 

4 mg/kg once, 4 mg/kg once daily for 7 days and then every 2 weeks until the first control mouse dies(died), or 4 mg/kg daily??

 

Firstly, let me say that the Baati study DID NOT use such a dose during the chronic toxicity arm. The 4 mg/kg arm(s) was used to see acute toxicity,  and affect of pre-administration on CCl4 challenge.

 

Secondly, If you are not replicating the 1.7 mg/kg dosing regime, the way Baati executed his study -- how can you expect your results to be comparable?

 

Thirdly, No human, that I know of, has taken a single 4 mg/kg dose ( that is a 380 mg dose for me @ .8 mg/ml saturation that is an entire pint of olive oil - 475 ml)  much less once daily for a week.

 

Even when Anthony Loera drinks a cup that is at most 2.5 mg/kg , but I wager a cup is approximate and his weight brings it to the 1.7 mg/kg used in the study.

 

Lastly, If you are using 4 mg/kg daily -- you may be proving chronic toxicity of such a dose. At least in regular lab mice ( I assume that for the longevity arm you are not using  NOD/SCID mice)

 

Note: 4 mg/kg daily for the average 80 kilo human would end up costing $4000-$8000 a year in C60 depending on purity and a couple thousand more in olive oil

Edited by sensei, 28 December 2015 - 11:08 PM.

[stefan_001]

I am simply stating that treatment with a C60oo product of one particular vendor is showing a strong trend of increased cancer burden in our xenograft model, and is also showing a strong trend of increased all-cause mortality in three ongoing studies. I make no further claims than that.

Thanks for sharing, I think I will put on hold considerations for own use or recommendation for others till it is clearer where this goes wrong. It also makes one wonder how to ensure that you buy quality goods....perhaps I need to build my own test and validation lab...

Edited by stefan_001, 28 December 2015 - 11:15 PM.

[kmoody]

Can you be a bit more clear?

 

4 mg/kg once, 4 mg/kg once daily for 7 days and then every 2 weeks until the first control mouse dies(died), or 4 mg/kg daily??

 

Firstly, let me say that the Baati study DID NOT use such a dose during the chronic toxicity arm. The 4 mg/kg arm(s) was used to see acute toxicity,  and affect of pre-administration on CCl4 challenge.

 

Secondly, If you are not replicating the 1.7 mg/kg dosing regime, the way Baati executed his study -- how can you expect your results to be comparable?

 

Thirdly, No human, that I know of, has taken a single 4 mg/kg dose ( that is a 380 mg dose for me @ .8 mg/ml saturation that is an entire pint of olive oil - 475 ml)  much less once daily for a week.

 

Even when Anthony Loera drinks a cup that is at most 2.5 mg/kg , but I wager a cup is approximate and his weight brings it to the 1.7 mg/kg used in the study.

 

Lastly, If you are using 4 mg/kg daily -- you may be proving chronic toxicity of such a dose. At least in regular lab mice ( I assume that for the longevity arm you are not using  NOD/SCID mice)

 

Note: 4 mg/kg daily for the average 80 kilo human would end up costing $4000-$8000 a year in C60 depending on purity and a couple thousand more in olive oil

 

Sure, happy to clarify.

 

This study design was based on an expected consumption of 4mg/kg C60oo per mouse over the course of a week. The measurements were based on a fixed average mouse mass and a fixed food consumption so there will be variance in the actual amount consumed (our current estimates are ~2.5 - 3 mg/kg). We are tracking food consumption and mouse weights so we can take these variables into consideration at the conclusion of the study.

 

I am not attempting to replicate Baati's study nor compare results per se. Baati's was not designed to be a proper lifespan study. Cutting off administration of C60 was a "the chronic toxicity study is over, lets just see what happens" sort of a situation, rather than something deliberately planned. It just happened to show an apparent lifespan effect. If there was even a small amount of chronic toxicity it is highly unlikely that the positive results Baati reported would have been observed. The modest study design changes (no loading dose, moderate chronic administration) in our study does not reasonably account for the variance in lifespan outcomes we are seeing, at least not in my opinion. You and others here are free to disagree. I'll just stick to reporting on the data.

 

In terms of study design, ours is statistically powered with proper group size (n=45 per arm), with a few critical changes. Our sponsor wanted to look at mice that were already middle aged so we used pre-aged C57BL/6 males for convenience. We are aware that there are limitations associated with this model but it was chosen for practical reasons. Likewise, we are administering via diet approximations rather than gavage. While less accurate, I feel the study size overcomes the assumed variance, especially considering we can adjust at the conclusion of the study because we have been tracking mouse mass and food consumption.

 

I have no input on rates of human consumption or cost of treatment at given doses.

 

So sure, I'll acknowledge that it is possible that what we are seeing is chronic toxicity. However, I feel that this is highly unlikely given the results reported by others and doses/methods of administration/strains in question. When taken with our other data it is very strongly suggestive of a product issue. If the all-cause mortality outcomes of the AML study oppose that of our pilot, that will put this issue to rest in my mind. I'm just waiting to see what happens there.

 

In the context of the discussion regarding prophylactic cancer treatment, it seems unlikely that one dose would magically eliminate all tumors and double lifespan, and our modestly different treatment regimen would cause premature mortality including deaths from cancer. Each argument as a stand alone is plausible I suppose, but collectively this simply does not add up.

[kmoody]

 

I am simply stating that treatment with a C60oo product of one particular vendor is showing a strong trend of increased cancer burden in our xenograft model, and is also showing a strong trend of increased all-cause mortality in three ongoing studies. I make no further claims than that.
 

Thanks for sharing, I think I will put on hold considerations for own use or recommendation for others till it is clearer where this goes wrong. It also makes one wonder how to ensure that you buy quality goods....perhaps I need to build my own test and validation lab...

 

Or source from a provider who manufactures under GMP. This is an area we are considering getting into for exactly these reasons. The GMP standard is a righteous pain to produce under (and by extension more expensive to buy), but you know that what is in the bottle has been well characterized and follows the highest degree of quality assurance.

 

But yes, I definitely agree with you.

Edited by kmoody, 28 December 2015 - 11:49 PM.

[sensei]

Sure, happy to clarify.

 

 

This study design was based on an expected consumption of 4mg/kg C60oo per mouse over the course of a week. The measurements were based on a fixed average mouse mass and a fixed food consumption so there will be variance in the actual amount consumed (our current estimates are ~2.5 - 3 mg/kg). We are tracking food consumption and mouse weights so we can take these variables into consideration at the conclusion of the study.

 

 

 

Thank you for the clarification.

 

Without replicating the Baati study with respect to dose and length of dose, any results regarding longevity are (to use 2 British terms) bolloks and rubbish.

 

Furthermore the strain of mouse you use [C57BL/6]is not particularly suited for longevity analysis as it

 

1. develops tumors on a fatty diet

2. prefers to drink ethanol [not a normal mouse]

3. tends to diabetes

4. mean study lifespans range from 600 days to almost 900 days with a huge standard deviation

 

Also, the tumor development rate at death is 70%;   cancer as a cause of death [because of C60] would only be [potentially] statistically significant if every single mouse died OF CANCER in  more than 1 iteration of the study.

 

http://www.informati...ocs/C57BL.shtml

Edited by sensei, 29 December 2015 - 12:57 AM.

[kmoody]

 

Sure, happy to clarify.

 

 

This study design was based on an expected consumption of 4mg/kg C60oo per mouse over the course of a week. The measurements were based on a fixed average mouse mass and a fixed food consumption so there will be variance in the actual amount consumed (our current estimates are ~2.5 - 3 mg/kg). We are tracking food consumption and mouse weights so we can take these variables into consideration at the conclusion of the study.

 

 

 

Thank you for the clarification.

 

Without replicating the Baati study with respect to dose and length of dose, any results regarding longevity are (to use 2 British terms) bolloks and rubbish.

 

Furthermore the strain of mouse you use [C57BL/6]is not particularly suited for longevity analysis as it

 

1. develops tumors on a fatty diet

2. prefers to drink ethanol [not a normal mouse]

3. tends to diabetes

4. mean study lifespans range from 600 days to almost 900 days with a huge standard deviation

 

Also, the tumor development rate at death is 70%;   cancer as a cause of death [because of C60] would only be [potentially] statistically significant if every single mouse died OF CANCER in  more than 1 iteration of the study.

 

http://www.informati...ocs/C57BL.shtml

 

Bollocks and rubbish constitute an unreasonable assessment. We'll have to agree to disagree on this. A "lifespan study" with n=18 (n=6 per group), as Baati's was, is not valid nor admissible as a lifespan by any stretch of the imagination. I do not contest that the Baati study was well done, but as a toxicity assessment, not a lifespan study. But to each his/her own.

 

That said, I am not suggesting that the lifespan study is an end all be all. I am simply suggesting, based on all available data, that there is a reasonable (in my mind likely) potential for C60oo sourcing to be a serious confounding variable. On what basis do you think this is an unreasonable hypothesis? Or are you simply emphasizing other possibilities in the interest of thoroughness?

Edited by kmoody, 29 December 2015 - 02:33 AM.

[sensei]

Bollocks and rubbish constitute an unreasonable assessment. We'll have to agree to disagree on this. A "lifespan study" with n=18 (n=6 per group), as Baati's was, is not valid nor admissible as a lifespan by any stretch of the imagination. I do not contest that the Baati study was well done, but as a toxicity assessment, not a lifespan study. But to each his/her own.

 

 

That said, I am not suggesting that the lifespan study is an end all be all. I am simply suggesting, based on all available data, that there is a reasonable (in my mind likely) potential for C60oo sourcing to be a serious confounding variable. On what basis do you think this is an unreasonable hypothesis? Or are you simply emphasizing other possibilities in the interest of thoroughness?

 

 

Firstly, the Baati Study was about chronic toxicity, not lifespan.

 

Secondly, the FACT that every single individual in the Baati n=6 cohort with C60OO EXCEEDED the mean and 3 SD of a normal wistar rat makes it statistically significant.

 

Thirdly, your design of experiment is severely flawed re: lifespan as it does not replicate the conditions that resulted in the 90% increase in lifespan seen in Baati.  The dosage and number of doses are not equivalent.

 

Fourthly, your choice of mouse is not optimal to studying lifespan due to the huge range of mean and SD regarding the lifespan of the strain.

 

Fifthly, your choice of mouse is not optimal due to issues regarding metabolic syndrome (predisposed to diabetes) and the 70% tumors at death.

 

I would posit that the single 4 mg/kg dose of C60OO has no effect on the ACTUAL all cause mortality rates of the mice in your study when compared to all cause mortality of the strain -- and will easily fall within a 2 SD statistical neighborhood of the mean, and be in-family.

Edited by sensei, 29 December 2015 - 02:55 AM.

[kmoody]

Firstly, the Baati Study was about chronic toxicity, not lifespan.

 

 

 

Secondly, the FACT that every single individual in the Baati n=6 cohort with C60OO EXCEEDED the mean and 3 SD of a normal wistar rat makes it statistically significant.

 

Thirdly, your design of experiment is severely flawed re: lifespan as it does not replicate the conditions that resulted in the 90% increase in lifespan seen in Baati.  The dosage and number of doses are not equivalent.

 

Fourthly, your choice of mouse is not optimal to studying lifespan due to the huge range of mean and SD regarding the lifespan of the strain.

 

Fifthly, your choice of mouse is not optimal due to issues regarding metabolic syndrome (predisposed to diabetes) and the 70% tumors at death.

 

I would posit that the single 4 mg/kg dose of C60OO has no effect on the ACTUAL all cause mortality rates of the mice in your study when compared to all cause mortality of the strain -- and will easily fall within a 2 SD statistical neighborhood of the mean, and be in-family.

 

I'm not quite sure what point you're trying to make here... I am not attempting to replicate the Baati study. If I were, I would replicate the Baati study. Our study is attempting to determine whether ~4mg/kg weekly dose administered by diet can increase the lifespan in pre-aged C57BL/6 males. The Baati study is irrelevant with respect to answering this question. You are entitled to your opinion regarding the merits of a C57BL/6 model vs. others. We discussed these and other issues in full with the study sponsor to arrive at our current study design. We are very familiar with the limitations. I think you exaggerate them but I am not in the business of this sort of speculation so I'll leave that to you. I am in the business of determining how available data should dictate future research. When I see relatively minor differences among studies and no consistency in outcomes, even at a trend level, and the one unifying variable is a sourcing variable, that suggests very obviously to me that the first place to look is sourcing. That is not to necessarily suggest it is the definitive cause, merely that it is the best place to start looking.

 

All that aside, we are seeing increased all-cause mortality from C60oo in all three studies. Would it not be consistent with your interpretation (that C60oo is awesome health-wise) that product sourcing could be the issue with the results we are seeing, rather than minutia between study designs? I mean, I run nearly identical xenograft studies, one doubles the lifespan and one doubles tumor burden. To me, this is highly suggestive of product sourcing being the issue and not some other variable. Why are you so quick to blame study design when a far more likely conclusion is that product sourcing is the problem? You make your stuff in house, which per my data should give you positive health outcomes. Everything I am seeing is consistent with your interpretation. Why are you so quick to discard our lifespan study?

 

I'm also note sure what you're getting at with the single 4mg/kg dose of C60oo. This is not what we did. For the lifespan study, mice are receiving approximately 4mg/kg C60oo per week for several months, not just once.

Edited by kmoody, 29 December 2015 - 03:24 AM.

[aconita]

Another thing about sonification is that the transducer wears off as well as the container does, those micro particles in C60oo may reach places usually they don't (mitochondrial membranes?) and even if in very tiny amounts may cause issues or at least change outcomes.

 

We don't know the nature of the transducer used as well as the nature of the container, nor we know the frequency, power, temperature and time, those are all variables too.

 

I think we already have enough to deal with and we don't really need more variables to add in.

 

Sonification of C60oo sounds like a great idea but at a closer look I think is not a smart choice.

 

Data coming from research done with sonificated C60oo should be excluded from evaluation because it is a different compound that invalidates the results.