Author Topic: Prostate cancer patient used to create 22Rv1 WAS infected with XMRV!!!!!!!!!  (Read 1418 times)

Tango

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Permission to repost!!!

Thank you Cath and Gerwyn.



How do we know that the patient that was used to create the 22Rv1 cell line was infected with XMRV?

The Coffin paper from Science last weeks shows that he was infect!!!

Here are some extracts from Paprotka et al.  As you can see their PCR assay could not amplify mouse ERVs or HERVs.

Quote
To quantify the amount of XMRV DNA in the CWR22 xenografts, we developed a real-time PCR primer-probe set that specifically detected XMRV env and excluded murine endogenous proviruses present in BALB/c and NIH3T3 genomic DNA (Fig. 1C).  We used quantitative PCR of 22Rv1 DNA to estimate 20 proviruses/cell and used the 22Rv1 DNA to generate a standard curve. The CWR22 xenografts had significantly fewer copies of XMRV env (<1–3 copies/100 cells) compared to the 22Rv1 cells (2000 copies/100 cells).  The CWR-R1 cell line had 􀗽3000 copies/100 cells, and the NU/NU and Hsd nude mice, thought to have been used to passage the CWR22 xenograft, had 58 and 68 copies/100 cells, respectively.


In early passage they only found XMRV env sequences.


Quote
We used the same XMRV-specific primer sets to amplify and sequence DNA from early passage xenografts (736, 777, 8L, 8R, 16R, and 18R; Fig. 2B); the results showed that XMRV env, but not gag sequences were present (sequencing coverage summarized in fig. S3), indicating that the early xenografts did not contain XMRV.


They say in the supporting material that:


Quote
Since no XMRV was detected in any of the early xenografts, the XMRV env sequences detected are from the endogenous Prexmrv1 provirus in the nude mouse DNA.


But look at the env section of PreXMRV-1.  It is only 91% similar to XMRV in some sections and thus would not be the same env.


Thus, the only place that the env sequences could have come from, is the prostate cancer sufferer whose cells were used to create the xenograft. 
« Last Edit: June 12, 2011, 09:45:55 PM by v99 »
"I suspect there have been a number of conspiracies that never were described or leaked out. But I suspect none of the magnitude and sweep of Watergate." Woodward

"I would favor any name that does not impose (or give the appearance of imposing) taxonomic preconceptions on the nomenclature." Coffin

Tango

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Translation:

Coffin went looking for the source of XMRV in a prostate cancer cell line, called 22Rv1.

There are several generations of this cell line that still exist.  So you can look in each one to see if XMRV is there.  If it was not there in the earlier generations, XMRV must have been created during the time the cells were taken from the prostate cancer patient and today.  This is not the same as using a test that cannot find XMRV in the earlier generations and is on the assumption that all tests work perfectly.

So when they looked all they could find was part of XMRV in the earlier generations. 

But as their test won't detect mouse endogenous viruses, won't find human endogenous viruses, and because this section of the virus does not match those viruses they found and named PreXMRV-1 and 2, it must be XMRV they found.

Which could only have come from the infected human.
"I suspect there have been a number of conspiracies that never were described or leaked out. But I suspect none of the magnitude and sweep of Watergate." Woodward

"I would favor any name that does not impose (or give the appearance of imposing) taxonomic preconceptions on the nomenclature." Coffin

Gerwyn

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Permission to repost!!!

Thank you Cath and Gerwyn.



How do we know that the patient that was used to create the 22Rv1 cell line was infected with XMRV?

The Coffin paper from Science last weeks shows that he was infect!!!

Here are some extracts from Paprotka et al.  As you can see their PCR assay could not amplify mouse ERVs or HERVs.


In early passage they only found XMRV env sequences.



They say in the supporting material that:



But look at the env section of PreXMRV-1.  It is only 91% similar to XMRV in some sections and thus would not be the same env.


Thus, the only place that the env sequences could have come from, is the prostate cancer sufferer whose cells were used to create the xenograft.

nothing to do with me   credit goes to you and cath

Tango

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"I suspect there have been a number of conspiracies that never were described or leaked out. But I suspect none of the magnitude and sweep of Watergate." Woodward

"I would favor any name that does not impose (or give the appearance of imposing) taxonomic preconceptions on the nomenclature." Coffin

Robyn

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So Coffin's test might not be able to find XMRV in the earlier cell lines because it is not sensitive enough?
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Tango

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No.  He found XMRV in the earlier cell lines.  The env region of XMRV, not a PreXMRV.
"I suspect there have been a number of conspiracies that never were described or leaked out. But I suspect none of the magnitude and sweep of Watergate." Woodward

"I would favor any name that does not impose (or give the appearance of imposing) taxonomic preconceptions on the nomenclature." Coffin

Dr. Yes

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V, can you set up a comparison between the env/gag primers used by Coffin and the appropriate env and gag sequences on preXMRV1?  I'd like to see if they are in a section with enough variation to throw off a PCR. 

[Btw the other possible source of XMRV in these experiments - well, marginally possible - is contamination that Coffin and co. accidentally introduced into their samples.  But I would consider the hypothesis of the patient as the source far more likely..]
« Last Edit: June 13, 2011, 07:06:40 AM by Dr. Yes »
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JT1024

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While I'm not online enough to read and absorb all the info, my brief encounters so far indicate there is so much more dirt to find!

Thanks V, Cath, Ger, and Doc for your expertise and persistence.  Wish I could contribute more to the hunt!

I was just watching a CNN "documentary" on Julian Assange.  I didn't watch the whole thing but it appears some of his (and his groups) talents could have been used to unearth information on XMRV and ME/CFS.  Like the character in Stieg Larsson's novels, Lizbeth Salander, Julian Assange is an infamous hacker. Those MRC files in the UK that are "secret" until the current generation is dead... would most likely be a piece of cake to him and his associates.  Similar files from the CDC and other US government agencies would be equally accessible.

Many of you know of my Christian beliefs and that I often do not concur with other members beliefs and opinions.  The reason I bring this up is my conscience.  I want to get to the bottom of this crap as much as anyone else does. I am not one to combat government agencies  since I am barely able to work.  I believe medical knowledge and research has been hindered or outright obstructed (unless I find information that proves otherwise).

It seems too many scientists lack a conscience and are playing games (or essentially recreating the scientific process to meet their agendas).  Gerwyn and others have stated how the "scientific process" has not been followed and I totally agree. Having worked in a clinical laboratory for many years, this is such a blatant disregard to science it should be "blasphemy" to any true scientist. Now there is an appropriate word to use against the "scientists behaving badly"!  They've abandoned their own fundamentals and other scientists should be calling them on it.

Where is Dr. Collins of the NIH? What are Drs. Alter and Lo saying to the likes of Coffin, Singh, Levy, etc.??

Too tired to continue further.

I look forward to more revelations. NOTHING will surprise me at this point.

First they Ignore you , then they Laugh at you , then they Attack you , then you WIN!!!

Mahatma Gandhi

hatshepsut

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Mark3981

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It seems too many scientists lack a conscience and are playing games (or essentially recreating the scientific process to meet their agendas).  Gerwyn and others have stated how the "scientific process" has not been followed and I totally agree. Having worked in a clinical laboratory for many years, this is such a blatant disregard to science it should be "blasphemy" to any true scientist. Now there is an appropriate word to use against the "scientists behaving badly"!  They've abandoned their own fundamentals and other scientists should be calling them on it.


Thanks JT. Frustrating to me as well.

It has to be miserable for the true scientists who have to deal with the double standard. Meaning the true scientists have to work with science and limited funding or else they loose credibility and face a barrage of attacks.

 THen we have the other side who seems to have enough money to buy, craft whatever opinion they want through the use of SPIN organizations, corporate owned media propaganda, faux advocacy organizations, political influence, and government officials beholden to the special interests. AND everything else I forgot to mention!!! whew.

What a dang mess. What a double standard!