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Product B - Telomerase Activation


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#391 Louis

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Posted 20 January 2012 - 02:17 AM

But Andrews has gone on public record in the past and stated that Sierra Sciences has tested TA-65 and that it does very weakly activate telomerase in their fibroblast screens. (Despite his more recent ambiguous answers to this question.)

It's cycloastragenol that's in question here.
Despite the fact that they test out very close on mass spect, there still appear to be some important differences.


Waitaminute... cycloastragenol and TA65 show a difference on mass spec? As in they are not structurally identical? The RevGenetics analysis was HPLC, wasn't it? The physical form, micronized or not, shouldn't matter in either MS, LC, or cell based assays, since it will be put into solution, probably in DMSO, in preparation for any of these.


Maybe I wasn't clear the way I wrote it. Cyclo and TA-65 test "very close on mass spect".
But I do not believe that TA-65 is simple bulk cycloastragenol.

#392 Louis

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Posted 20 January 2012 - 02:52 AM

Thanks, Anthony. Is this the one you're referring to? Perhaps someone here might explain the significance of "the transcription of human telomerase reverse transcriptase gene" and how it relates to protecting or lengthening telomeres.

http://www.ncbi.nlm....pubmed/22197555

SIRT1 prevents replicative senescence of normal human umbilical cord fibroblast through potentiating the transcription of human telomerase reverse transcriptase gene.
Yamashita S, Ogawa K, Ikei T, Udono M, Fujiki T, Katakura Y.

Abstract
SIRT1, the mammalian homolog of sirtuins, has emerged as a mediator of the beneficial effects of calorie restriction. Among them, we focused on the SIRT1-induced prevention of cellular senescence, and tried to reveal the molecular mechanisms that define the effects of SIRT1. Firstly in this study, we observed that overexpression of SIRT1 resulted in the prevention of cellular senescence of normal human umbilical cord fibroblast HUC-F2 cells. Here, we focused on the human telomerase reverse transcriptase (hTERT) gene as a target of the SIRT1-induced prevention of cellular senescence. Results showed that SIRT1, SIRT1 activator, resveratrol, and SIRT1 activating condition, starved condition, increased the transcription of hTERT in HUC-F2 cells. Next, we found that SIRT1 increased hTERT transcription in a c-MYC-dependent manner, triggered the transcription of the c-MYC gene and increased the amount of c-MYC recruited to the hTERT promoter. Further, SIRT1 increased the transcriptional activation ability of c-MYC and correspondingly increased the amount of acetylated H4 histone at the hTERT promoter. All of these results indicated that SIRT1 activates hTERT transcription through the involvement of c-MYC, and suggested that this SIRT1-induced augmentation of hTERT transcription resulted in the extension of the cellular life span of HUC-F2 cells.

Transcription means that an RNA copy of genomic DNA is being created. This report probably does not mean that taking resveratrol with lengthen your telomeres. You have to look at the dose they used in their in vitro system, and see if it's even possible to attain that level in vivo. Usually in this kind of study, it isn't.


For most cells, transcription of hTERT is the rate limiting step in producing the telomerase enzyme. So these cells should be producing more telomerase enzyme w/ resv than without, assuming that the resv is in fact activating SIRT1. The authors are trying to argue that this is why the cells are living longer with resv in the experiment.

But like cycloastragenol which operates on hTERT very indirectly by phosphorylating ERK, resveratrol appears to be operating in this experiment through another indirect mechanism involving first SIRT1 and then c-MYC to ultimately affect hTERT. This mechanism is again naturally present in the cell and it looks like the key starting point to activate it is SIRT1. The resveratrol is not directly interacting with the hTERT repressor as you might expect an activator to do; it's working through this indirect mechanism involving sirtuins to ultimately get to hTERT.

I haven't read the full paper, but the authors appear to be assembling the beginnings of a bigger argument that calorie restriction and telomerase are related through SIRT1 and then c-MYC. I think they are putting together a case that restricting calories activates this indirect SIRT1/c-MYC mechanism to indirectly turn on hTERT and produce telomerase.

Very very interesting abstract. I was not aware that there was any connection between calories restriction and telomerase, so this is pretty exciting to me -- even though it's just one little isolated experiment.

Edited by Louis, 20 January 2012 - 03:00 AM.


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#393 Louis

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Posted 20 January 2012 - 06:02 PM

Another study showing a connection between calorie restriction and telomerase in vitro:
http://www.fasebj.or...24/5/1442.short

In a healthy cell line, restricting glucose in the experiment activates telomerase. In a cancer cell line, it de-activates telomerase. Again, an example where the mechanisms controlling telomerase in healthy vs cancer cells are completely different.

#394 AdamI

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Posted 20 January 2012 - 06:28 PM

Another study showing a connection between calorie restriction and telomerase in vitro:
http://www.fasebj.or...24/5/1442.short

In a healthy cell line, restricting glucose in the experiment activates telomerase. In a cancer cell line, it de-activates telomerase. Again, an example where the mechanisms controlling telomerase in healthy vs cancer cells are completely different.


Interesting, soo one should try to starve during the sessions of Cyclo & AIV and other telomerase activators.

#395 hav

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Posted 20 January 2012 - 11:02 PM

I had a negatjve experience once taking a gram of astragalus root on an empty stomach. Probably more optimal to take with a high fiber cereal or low fat yogurt.

Howard

#396 Louis

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Posted 23 January 2012 - 08:33 PM

Hot off the presses, just published Jan 11, 2012.
This observational study in humans finds an inverse correlation between calorie intake and telomere length:
http://www.ncbi.nlm....pubmed/22237065

This is the strongest evidence of a link between telomeres and calorie restriction that I've been able to find, and that's not saying much given the observational nature of the study.
The total evidence to date is very weak, but nevertheless intriguing.

Edited by Louis, 23 January 2012 - 08:39 PM.


#397 Anthony_Loera

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Posted 26 January 2012 - 02:43 PM

But Andrews has gone on public record in the past and stated that Sierra Sciences has tested TA-65 and that it does very weakly activate telomerase in their fibroblast screens. (Despite his more recent ambiguous answers to this question.)

It's cycloastragenol that's in question here.
Despite the fact that they test out very close on mass spect, there still appear to be some important differences.


Waitaminute... cycloastragenol and TA65 show a difference on mass spec? As in they are not structurally identical? The RevGenetics analysis was HPLC, wasn't it? The physical form, micronized or not, shouldn't matter in either MS, LC, or cell based assays, since it will be put into solution, probably in DMSO, in preparation for any of these.



You got a point niner.

Lets remember, Dalton in Canada confirmed NMR cycloastragenol content data in astragalus samples we sent way back in 2008. They used (HPLC-ELSD, NMR and MS). This data was then sent to two labs... Intertek AACL and Chromadex. Samples of different cycloastragenol material, were then sent to these labs to verify accuracy of new testing HPLC methods created from the Dalton information. After it was established that accuracy was solid, we started using them exclusively to verify cycloastragenol in all products we manufactured.

So why is Dalton lab so important? TA Sciences used it to verify that TA-65 was not found in regular off-the-shelf astragalus supplements. You can then see why we used them in our initial investigation regarding astragalus.

This is ancient history for the most part. Most of this information was posted here a loooooooonngg time ago (January 2009 apparently):
http://www.longecity...post__p__289595

Cheers
A

Edited by Anthony_Loera, 26 January 2012 - 02:46 PM.


#398 niner

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Posted 26 January 2012 - 04:22 PM

Anthony, I think that you have solid analytical work; HPLC, NMR, and MS are sufficient to nail down a structure as well as get a picture of the contaminants. I'm still not sure what Louis meant about "very close" on MS. Mass spectra have a binary nature; a peak that corresponds to a particular mass is either there or it isn't. The masses change by units of neutron/proton mass; that's what leads to the clean spectra.

#399 Louis

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Posted 26 January 2012 - 07:24 PM

The accuracy of mass spect in measuring molecular mass is usually only accurate to several ppm. Sometimes that's not enough.

#400 Louis

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Posted 26 January 2012 - 07:57 PM

Does someone have chromatography and NMR data specifically on TA-65?

That might help explain why they behave differently in the Sierra fibroblast screen.

#401 Louis

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Posted 26 January 2012 - 09:52 PM

Here we report initial findings from a dietary supplement
program which includes TA-65, a purified smallmolecule
telomerase activator derived from an extract of a
plant commonly used in traditional Chinese medicine.
Telomerase activation and functional studies on a related
molecule (TAT2) from the same plant have been previously
reported for human skin keratinocytes and immune cells in
culture.36 Effects of TAT2 in tissue culture studies with
CD8þ T cells from HIV/acquired immunodeficiency syndrome
(AIDS) subjects included increased replicative capacity,
improved cytokine and chemokine responses to
antigens, and increased killing of autologous HIV-infected
CD4þ cells.

--------

The above is quoted from the paper on the TA-65 human trial:
http://online.lieber...9/rej.2010.1085

Harley goes out of his way to specifically describe TAT2 (cyclo) as distinct from TA-65, but "related".

So is standard liquid chromatography mass spec really sufficient to identify the precise unique chemical structure of TA-65, or is it really only narrowing down the possibilities to a small subset of related compounds which include both TAT2 and TA-65. Sometimes you just can't be 100% sure you have the unique structure from standard LC-MS alone.

That's why I'm saying they test very close (which yes is technically an abuse of the proper terminology), because I'm not convinced that standard LC-MS proved that the exact unique structure of TA-65 is the same as cyclo.

Additional tests like NMR would really help to settle it once and for all.

Edited by Louis, 26 January 2012 - 09:58 PM.


#402 PWAIN

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Posted 27 January 2012 - 12:59 AM

I thought there was a patent doco floating around a while back that specifically mentioned Cyclo in relation to TA-65.
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#403 Anthony_Loera

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Posted 27 January 2012 - 03:32 AM

How can identical twins not be related?

;)

Edited by Anthony_Loera, 27 January 2012 - 06:13 PM.


#404 AdamI

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Posted 27 January 2012 - 02:56 PM

The accuracy of mass spect in measuring molecular mass is usually only accurate to several ppm. Sometimes that's not enough.


All this talk about Mass spect, and I stumbled on this, some new software for analyzing with that instrument, if someone is interested:
http://www.spectrolyzer.com/home/

#405 niner

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Posted 27 January 2012 - 03:34 PM

The accuracy of mass spect in measuring molecular mass is usually only accurate to several ppm. Sometimes that's not enough.


It depends on the exact technique; there are ways to see changes in a single proton on a protein, for example, though not all MS methods would be appropriate for that. For small molecules like cycloastragenol, MS should have no problem at all in pinning down the exact mass of the parent ion and any fragments; it's a fairly easy problem when something is that small.

#406 Louis

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Posted 27 January 2012 - 04:36 PM

The rule of thumb I have in my head is: With 5ppm accuracy, you can uniquely determine a formula for molecular masses below 200u.
I believe cyclo has more than twice that mass. So you'd need a pretty high accuracy setup.

Half the time it's just tech's that run these things and then they're interpreted purely by software. And for the cheap tests, they tend to use the cheap equipment.

Has anyone run NMR on TA-65?

#407 niner

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Posted 27 January 2012 - 07:30 PM

The rule of thumb I have in my head is: With 5ppm accuracy, you can uniquely determine a formula for molecular masses below 200u.
I believe cyclo has more than twice that mass. So you'd need a pretty high accuracy setup.

Half the time it's just tech's that run these things and then they're interpreted purely by software. And for the cheap tests, they tend to use the cheap equipment.

Has anyone run NMR on TA-65?


Cyclo MW is 491; You would need a very good setup to get the parent ion with absolutely no ambiguity, but I'm presuming a fragmentation analysis, which could be done on a lower resolution instrument. Cyclo isn't a simple molecule; in addition to its size, it has a dozen stereocenters. Anthony mentioned that Dalton ran an NMR.

#408 Louis

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Posted 27 January 2012 - 10:35 PM

You bring up a good point: I wonder if TA-65 is just a particular stereoisomer of cycloastragenol?
That would certainly explain the mass spec results. And it might explain biological differences in activating hTERT (e.g. why TA-65 works in the Sierra fibroblast screen, but bulk cyclo did not).

Did he say specifically that Dalton ran the NMR on TA-65? I thought he was referring to other astragalus compounds or cyclo w/r/t to Dalton tests.
You may be able to tell from NMR if TA-65 contains a higher percentage of a particular stereoisomer than bulk cyclo.

Edited by Louis, 27 January 2012 - 10:38 PM.


#409 Anthony_Loera

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Posted 30 January 2012 - 03:54 PM

Good news:
I was going over our internal standards regarding telomerase activators... and we have decided to move the needle.

What this means, is that although TA65 remains the standard to beat, we will begin to sell telomerase activators that do not meet the TA65 standard, but still show some activity (better than controls) at much higher concentrations.

This will allow RevGenetics to sell Product B.

I am awaiting on Louis to provide his link to "signup" so that he benefits. He took the most grief from my posts, and deserves (in my opinion) a token of confidence for putting up with me, and hanging in there.

IM me your link Louis.

Cheers
A


#410 Louis

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Posted 30 January 2012 - 04:03 PM

Anthony, I don't understand.

In particular, I'm not clear on what you're asking me to do here or to send you?

#411 Anthony_Loera

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Posted 30 January 2012 - 04:55 PM

Good news:
I was going over our internal standards regarding telomerase activators... and we have decided to move the needle.

What this means, is that although TA65 remains the standard to beat, we will begin to sell telomerase activators that do not meet the TA65 standard, but still show some activity (better than controls) at much higher concentrations.

This will allow RevGenetics to sell Product B.

I am awaiting on Louis to provide his link to "signup" so that he benefits. He took the most grief from my posts, and deserves (in my opinion) a token of confidence for putting up with me, and hanging in there.

IM me your link Louis.

Cheers
A



Louis, you are an independent vendor or member that buys Product B according to an old post of yours, so you must have a member name that you signed up under. If you are a member, then you will also have a link like this one... except, your member name would go where the XXXXXXX go in the URL below:

http://XXXXXXXX.<spa...ml/rel=nofollow

IM me your member name, or the link you where given to refer folks.

A

#412 Louis

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Posted 30 January 2012 - 05:31 PM

I understand what you're saying now Anthony.

If I were to give you my link/referral number and you were to use it in signing up with Isagenix, I would then derive a profit from your sales of Product B. That's how the MLM model works.
I do appreciate the nice gesture. But I do not currently derive any profit from the sale of product B (aside from what I sell only to myself), and I specifically do NOT want to derive any profit from it. I feel that would be a conflict of interest.

I only buy product B because of the results I've had and to support future research at Sierra Sciences. I have no personal interest in making money from it.

But again, I appreciate the gesture. It's nice of you.

Here's what I suggest you do. Use the Bill Andrews / SierraSciences link when you sign up:
http://telomere.isag...s/en/home.dhtml

This is the account / website specifically set up by Bill Andrews and Sierra Sciences. He has pledged that any profit he derives from product B sales using this site will be funneled directly back into research at Sierra Sciences. If you sign up using Bill Andrew's link, then every time you sell a bottle of product B the proceeds will go directly to Sierra Sciences instead of going to someone such as myself or some random distributor of the product.

By doing this, you will be greatly helping the overall cause of life extension in general and the work that Sierra Sciences is doing with every sale of the product.
That would be a wonderful gesture for everyone.

Louis

Edited by Louis, 30 January 2012 - 05:36 PM.

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#413 Anthony_Loera

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Posted 30 January 2012 - 05:40 PM

Hi Louis,

I wanted you to IM this infor to me, as I am not asking people to sign up. Many folks will not want to sign up, and want to be able to purchase products without the hassle of possible autoship, which the MLM signup sometimes creates.

Are you sure you aren't Bill? :)

Cheers

A

Edited by Anthony_Loera, 30 January 2012 - 05:42 PM.


#414 Louis

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Posted 30 January 2012 - 06:08 PM

Anthony, I have never personally spoken to or met Bill Andrews.
I'm quite certain he has absolutely no idea who I am.

He gave away the link to his website/account above during one of his public interviews. You can find that interview online. I'm just passing it on. It's public information.

#415 Anthony_Loera

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Posted 30 January 2012 - 06:18 PM

That is fine, thanks.

I am looking at the extensive policies and procedures for signing up. It appears I have run into a snag, as internet sales using resale inventory on non-isagenix sites maybe an issue for retailers and possibly not-allowed. I have called, and the question has caused them to punt, and send my question to their compliance area... this run around is proving quite educational, as I can see the limited sales structure this MLM may have... and why you may never see this product in regular retail or various website stores along with your favorite supplements and products.

Well, let's see what compliance responds with. At main issue is the section termed 4.7 Resell of product

I also see lots of violations in this thread, regarding the policies and procedures... so I suppose it is good that no one here actually shares their membership name.

A

Update:

I received an auto response from compliance... hopefully the answer doesn't take 2-4 weeks as they state below, yikes:

Thank you for contacting our Isagenix Compliance Team via email. You will receive a response from one of our representatives within 24-48 hours.
However, all material submitted for review can take up to 2-4 weeks for an approval, so be sure to submit the request well in advance of the intended use. (This includes marketing materials, websites, books, ect)


Edited by Anthony_Loera, 30 January 2012 - 06:22 PM.


#416 Anthony_Loera

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Posted 30 January 2012 - 06:44 PM

Ahhh... I found a provision I assumed was there, but hoping it wasn't:

======================================================================

5.3 Cross Sponsoring/Other Business Opportunities

5.3.1 An Independent Associate or Preferred Customer shall not sell to other Isagenix Position Holders, non-Isagenix products or in any way promote to such Isagenix Position Holders the opportunities in marketing programs of other companies

5.3.2 You cannot sponsor or attempt to sponsor anyone from your personally enrolled or any other Isagenix sales group (downline or upline) into any other multi-level marketing (MLM) or network marketing organization. Nor may you introduce other business opportunities, goods, or services to any Isagenix Position Holder, including those you have personally enrolled. Violation of these policies is grounds for termination of the Independent Associate or Preferred Customer. It may also give rise to other claims for unauthorized use of our confidential information.

======================================================================

Sounds like no one could offer better products to associate folks, even if they are proven better...

So Louis, even if I became a wholesaler, and sold 3 different telomerase activator products, there is no way I could offer you say... Astral Fruit or TA-65, even if it was proven better than Product B... simply because you (and I, if I signed up) would be bound by this policy. The good part is that it doesn't apply to folks who are not members, but only buy retail from my shop.

Maybe I can live with that... hmmm

A

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#417 Anthony_Loera

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Posted 30 January 2012 - 07:06 PM

I don't know.... here are some more limitations:

============================================================================

6.2 Internet and Web Site Policy

6.2.1 Isagenix maintains an official corporate Website and provides Web sites to all active-status Independent Associates and Preferred Customers. All Independent Associates and Preferred Customers must use the Isagenix-provided Web sites only. Third-party Web sites will not be allowed. An Independent Associate may advertise on the Internet by placing a pre-approved “Banner image” on Web sites that link directly to their Isagenix-provided Web site. You may only use your Isagenix-provided Web site in connection with your Isagenix activities. All Independent Associates and Preferred Customers must implement and publish a privacy policy that protects any information gathered from being sold or used by anyone else.


6.5 Trade Shows
With our written authorization, you, as an Independent Associate, may present our products and income opportunity at trade shows. We must receive a written request for participation in trade shows at least two weeks prior to the show. Only Isagenix products and/or opportunity may be offered in the trade show booth. Only Isagenix produced marketing materials may be displayed or distributed. You may not sell or promote our products or income opportunity at informal and formal gatherings for the barter or sale of goods including but not limited to flea markets, swap meets, garage sales, or street markets.


6.8 Compliance Review

6.8.1 If an Independent Associate wishes to use a mode of advertising their Isagenix business outside of the approved marketing materials and replicated website the Independent Associate must receive written approval from the Isagenix Compliance Department prior to printing or posting such advertisement, copy or any other discussion or claims regarding Isagenix, the products or the income opportunity.

6.8.1.1 Modes of advertisement referred to in this section include but are not limited to: posting on MySpace, YouTube, eBay, or Craigslist, posting on Blogs and Forums, producing print and other multimedia marketing materials, and Team Building Websites.

6.8.2 The Independent Associate may only utilize such advertising medium when they receive written notification directly from the Isagenix Compliance Department.
============================================================================

This would make for a very slow marketing campaign, specially in these forums.

We will confirm section 6.2.1 above regarding 3rd party websites, as that one will obviously become a major issue should Product B actually come out with a 3rd version of the product, (that they say is much better than the current product b version they are selling to the public).

Having said everything, even if version 3 of Product B where to meet the TA-65 standards regarding telomerase activation, the legalese and limitations on this MLM product really appears to limit the sales of it. I don't think retailers would be able to sell this at all because of the policies. This is definitely not Avon.

I suppose I will simply wait on compliance to see what they say.

A

Edited by Anthony_Loera, 30 January 2012 - 07:10 PM.


#418 Anthony_Loera

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Posted 30 January 2012 - 07:56 PM

Well, compliance just emailed me back.

Sure enough they copy-pasted section 6.2 which states no third party sites. I suppose we didn't really need to lower our standards.

After all that difficulty, I am seriously happy that TA-65 remains the standard to beat. I am also happy that we can sell TA-65 to normal folks without asking them for their social security number and birthday, like the MLM for Product B does.

Cheers
A


#419 Logic

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Posted 01 February 2012 - 07:38 AM

The question of whether resveratrol, circumin etc. inhibit the expression of telomerase in non cancerous cells has been investigated by Vince here:

http://www.anti-agingfirewalls.com/2009/06/11/do-resveratrol-curcumin-and-egcg-from-green-tea-really-inhibit-the-expression-of-telomerase/
I would like to believe that nature is cleverer than all of us and that suppliments that inhibit telemorase expression in cancer cells dont do so in normal/healthy cells?
This certainly seems to be the case?

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#420 mdlee19

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Posted 03 February 2012 - 12:48 PM

Hi Anthony,
Since you placed your original order with me for Product B near the beginning of this thread, I will be happy to give folks a link to sign up with isagenix to get Product B at the member discount. My email is mdlee19 AT yahoo.com. I will reply with a signup link.
Thanks.

Mike


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