The intrepid Dr. Mikovits went up against Darth Vader today in Ottawa. In the face of incredible adversity, she took the heat. For us. And she points the way to the next step. Next Generation Sequencing. Here are her slides. Click each to enlarge. May the force be with her. Brava!
>Thank you, Dr. Deckoff-Jones. Wow! Thank you for the slides; these are really helpful.
I especially like the RMCP.
Patricia Carter
>Jamie: Was the last slide- the mouse contamination procedure- part of Judy's presentation or did you add it? Whatever- it's hilarious, but if Judy closed with it- more power to her. A light, humorous touch in the face of such adversity is very classy!
>my NK cell function went from single digits to 60 LU on HIV meds
>Thanks for being our Princess Leia!
>Scott,
The slide is Judy's. That's Max. We all owe Max a debt of gratitude.
And here's some more levity. Dr. Judy and Dr. Snyderman:):
http://www.youtube.com/watch?v=1ToztqqDcaY
Jamie
>Jamie: Wrong link. That one takes you to a trailer for an upcoming star wars movie- entertaining but no dr. judy :-)
>Right link. Sorry, Scott, my peculiar sense of humor:). Our heroes fighting to restore hope and peace across the galaxy:). I should have warned you.
Jamie
>What a waste of time and money this all is. Time to move on
>Why were the results yesterday publicized one day before the conference?
>To the Scientist that wants to move on. I am all for that. I have an HGRV, there is no other explanation for the improvement of my CFS and leukemia with ARVs. Furthermore, an early analysis of our survey suggests 1/4 partners catch CFS. Dr. Coffin, there is an HGRV in CFS patients. It is time to stop arguing about sequences, look at the clinical realities. Yes it is time to move on AND spend some money – to find a treatment.
Michael Snyderman, MD
>Mr/Mrs Scientist, in your view is a waste of time and money, in our view is a must that should have taken place 30 years ago with the first outbreaks of this illness.
We deserve some research in the end, We have lived with this long enough, it is about time that research is done on this field, and We all trust that the retroviral clues pointed by WPI makes a hell of a lot of sense in a chronic illness that started with an infection in most of the cases.
We all know this, because we all started to be sick the same way, out of a sudden we were sick!
A waste of time has been the 30 years that no public funding has been dedicated to solve ME/CFS.
A waste of money is the research done on CBT in the psychiatric field. Would not make sense for cancer or aids, what makes anyone think that it does make sense for ME/CFS?
You better think, and find out the real story of ME before you speak in here an easy comment like that.
>This must be a retrovirus, We all know because We all though that We had contracted HIV when we fell sick. It felt like aids, it felt like lack of immunity, like not being able to resolve whatever infection we got.
I still remember that in my worst days in the first 3 years, I was even scared to have sex with someone, because a sole kiss put me into beed for days, the minimal dose of bacteria or virus that came inside my body was a drama for my health, and I could feel it, I could feel how any "exposure" had an immediate effect on me and how my body responded.
That is why I am still convinced that regardless of the name, XMRV, PMRV, HGRV, BGRV, it will end up being a retrovirus that causes immune deficiency.
>From “The Demon-Haunted World: Science as a Candle in the Dark” by Carl Sagan, 1996.
If we resolutely refuse to acknowledge where we are liable to fall into error, then we can confidently expect that error — even serious error, profound mistakes — will be our companion forever.
There are no forbidden questions in science, no matters too sensitive or delicate to be probed, no sacred truths. That openness to new ideas, combined with the most rigorous, skeptical scrutiny of all ideas, sifts the wheat from he chaff. It makes no difference how smart, august or beloved you are. You must prove your case in the face of determined, expert criticism. Diversity and debate are valued. Opinions are encouraged to contend — substantively and in depth.
The British physicist Michael Faraday warned of the powerful temptation “to seek for such evidence and appearances as are in the favour of our desires, and to disregard those which oppose them….We receive as friendly that which agrees with [us], we resist with dislike that which opposes us, whereas the very reverse is required by every dictate of common sense.”
Valid criticism does you a favor.
The more we want it to be true, the more careful we have to be. No witness’s say-so is good enough. People make mistakes. People play practical jokes. People stretch the truth for money or attention or fame. People occasionally misunderstand what they’re seeing. People sometimes even see things that aren’t there.
What skeptical thinking boils down to is the means to construct, and to understand, a reasoned argument and — especially — to recognize a fallacious or fraudulent argument. The question is not whether we like the conclusion that emerges out of a train of reasoning, but whether the conclusion follows from the premise or starting point and whether that premise is true.
At the heart of science is an essential balance between two seemingly contradictory attitudes — an openness to new ideas, no matter how bizarre or counterintuitive, and the most ruthlessly skeptical scrutiny of all ideas, old and new. This is how deep truths are winnowed from deep nonsense. The collective enterprise of creative thinking and skeptical thinking, working together, keeps the field on track.
Nature is always more subtle, more intricate, more elegant than what we are able to imagine. And as in a detective story, it’s a joy to frame key questions, to work through alternative explanations, and maybe even to advance the process of scientific discovery.
*
I still think that my own experience with this disease, and what I know of others’ experiences with it, makes more sense with a retrovirus that acts in the way that I’ve been told that XMRV acts. But that is just a hypothesis.
If WPI wants to pursue this hypothesis, by all means they should do so. I wish them every success.
But what’s going on in this community right now has nothing in common with any sort of science I’ve ever heard of.
Science is pursued in the lab and through critical discussions with other individuals who care enough about getting to the truth to ruthlessly dig into the ideas — not by rallying the troops.
Cordially,
Lisa Petrison, Ph.D.
>That last slide was about the only one I understood. I needed the laugh. :)
>There are three flaws in the BWG study design and implementation that invalidates the results.
-In Lo/Alter's paper they used two PCR assays. For the sake of simplicity called 1 and 2. PCR 1 found NOTHING. In the BWG Lo used assay 1.
-Not all patients in the negative group were validated as being negative because not all patients in that group were tested.
-The patients in the positive group were on antiherpes and other medication which would have caused false negative results.
This borders on the negligent and who will take responsibility for wasting so much taxpayers money?
>This is getting embarrassing now. Let's just recap the results of the blood working group:
1– The labs that cant find XMRV in patient samples can find XMRV when it is there:
Of the nine labs that tested the samples, only one lab could not reliably detect XMRV in the positive controls (blank samples spiked with XMRV): The Whittemore Peterson Institute
2– Of the nine labs that tested the samples, only two found XMRV in any experimental samples (CFS and healthy controls):
WPI and NCI, the two groups that have published the 'XMRV–>CFS' Science paper.
3– WPI and NCI 'found' XMRV at equivalent rates in the negative controls and the CFS patients:
WPI called 8/15 negative controls 'positive'. They called 6/10 of their own 'positives' positives. They called 5/5 Lo et als positives positive.
NCI called 10/15 negative controls 'positive'. They called 5/10 WPIs positives positive. They called 2/5 Lo et als positives positive.
4– WPI and NCI 'positive' results did not correlate:
WPI called Negative Patient #10 positive. NCI called Patient 10 negative. NCI called Negative Patient #13 positive. WPI called Patient 13 negative. Over and over.
5– WPI and NCI contradicted their own results:
WPI Patient #2– WPI called this patient negative on 2/3 assays. Positive by one assay once, but not twice. NCI called Patient #2 negative on two assays.
WPI Patient #1– WPI called Patient #1 negative. NCI called them positive on one out of two assays. But the 'positive' result was only positive 1/2 times the procedure was replicated.
These results speak for themselves. My positive XMRV test is not worth the paper it is printed on. Unless the WPI get their act together, I think they may well be receiving a number of demands for refunds very soon. And what is going on with people blindly following Mikovits like she's the new messiah. She's sounding more and more unhinged. I can see this ending up with several hundred patients and Mikovits barricaded in the WPI and TV cameras and the army outside, a la Waco, Texas.
Mikovits has one more chance to redeem herself and she'd be advised to keep her head down and mouth shut until then. If the Lipkin study is a blowout, then she will have to go, if the WPI is ever going to rebuild their reputation, and I don't think the job offers will come flooding in for her.
Apologies for the undoubted offence, but sometimes these things have to be said. I hope I turn out to be "doubting Thomas", but I doubt it…
>The facts on the ground, the clinical realities speak for themselves. I have CFS and leukemia and both have responded to ARVs. An early analysis of our survey indeed sugests that CFS is infectious.
As physicians, we have to deal with labs that don't fit the clinical picture, both false positives and false negatives. We are trained to analyze the individual situations and make the best synthesis of all the data that we can. My best assessment is that there is an HGRV and that it can be treated and I am pleased with my response. If I had listened to all the negativity out there, I would have deprived myself of a response that has added at least a year to my survival.
Michael Snyderman,MD
>Michael, I'm delighted that you've responded so well to ARVs and I really hope you continue to do so. This could be an indication that you are infected with an HGRV, or it could equally be an indication that you are infected with another type of retrovirus, or the ARVs may be having some other type of therapeutic effect on your immune system, or some other unknown. It's quite possible you're experiencing a very real placebo effect or there are other unpredictable factors at play. I don't see why it has to be an HGRV. Also, many other "XMRV positives" haven't responded so well to ARVs.
ME/CFS may well be infectious, though a selective survey can't confirm that, but again that doesn't mean it's an HGRV. It could be another unidentified retrovirus or other pathogen. It could be that several pathogens can trigger the same disease. It could be a re-actived endogenous retrovirus. The truth is that these questions have not been answered and the WPI and Ruscetti's assays are completely unreliable.
I have good reason to believe in XMRV, not only because of my ME/CFS, but also my Father was diagnosed with prostate cancer earlier this year, but emotion and circumstance do not change facts. The BWG results could not have been any worse. There was no consistency at all. How can the original study results be taken seriously when the WPI's assays are so unreliable and with such a high false-positive rate. Even the discoverer of XMRV, Robert Silverman, is holding his hands up and admitting contamination with regard to both his involvement in the WPI study and his prostate cancer work. For the ME/CFS / HGRV connection to be real, it would take such a bizarre set of events to be true, though I admit stranger things have happened. We'll see.
Whatever happens, I hope you continue to go from strength to strength and we all start to see some light at the end of the tunnel.
Ed
>Here are three articles that have run in the past two days, criticizing the use of ARV's in CFS:
http://www.nytimes.com/2011/09/23/us/off-label-use-of-hiv-medications-is-catalyst-for-more-controversy.html?_r=1
http://www.fiercepharma.com/story/should-hiv-drugs-get-label-use-chronic-fatigue/2011-09-23#ixzz1YsX8qExh
http://www.thebody.com/content/64087/using-antiretrovirals-for-chronic-fatigue-syndrome.html
The first is from the New York Times. Perhaps even more disturbingly, the third is from a newsletter for AIDS patients.
This seems like it thus is becoming a PR problem for us. None of these articles exactly suggest that CFS patients aren't sick, but they certainly make it seem like it's more important that AIDS patients have access to the drugs than that CFS patients get to use them — especially since the retroviral connection to CFS has a lot less weight behind it now than it did a few days ago.
AIDS patients are extremely well-organized and vocal. I certainly do not want to become the focal point of their wrath!
Of course, it's not like if CFS patients weren't taking ARV's, they would be available to AIDS patients without insurance or money to pay for them. The funding comes from different sources. But that seems to be a point that, so far, has not prevented the articles from being written.
One point that I think we have in our favor is that Raltegravir is known to be effective against herpes viruses. Obviously there's substantial literature that shows that many CFS patients do indeed have reactivations of this class of virus, and it may be that Raltegravir is less toxic than Valcyte (used by doctors like Jose Montoya of Stanford) or other anti-herpes drugs such as Vistide (used by Peterson's clinic). So insofar as this is the argument for using Raltegravir, I don't think anyone can have any objections.
http://www.aidsmeds.com/articles/hiv_herpes_Isentress_1667_19150.shtml
The other drugs may have a wider spectrum of efficacy than has been currently researched. Tenofovir, for instance, is already known to have efficacy against Hepatitis B. I tend to think it would be a bit difficult to make an argument for CFS patients starting the drugs in the hope of randomly killing something (that would sort of be like throwing darts blindfolded). However, if people have had success on the drugs already, I think we can make a reasonable argument (even to people who have dismissed the idea that XMRV is a factor for us) that this is possibly due to their controlling one of the many infections that plague people with our disease rather than a placebo effect.
Best,
Lisa
>Ed:
You wrote: "It's quite possible you're experiencing a very real placebo effect…"
What very real mechanism do you propose quite possibly explains the data?
>Samuel,
Here's an article that suggests that EBV is a "progression factor" in CLL. Conceivably other herpes family viruses (including those that have yet to be discovered) could be be serving this role as well.
http://www.infectagentscancer.com/content/5/1/22
As I said earlier, the idea that an XMRV-like retrovirus is active in this illness is one that I think makes sense. But as scientists, being open to alternative explanations is the standard way to proceed when solid evidence is not yet in.
Best, Lisa
>Samuel, if I knew the mechanism by which the placebo effect worked, I'd be a rich man and possibly also a well man. However, no drug researcher and very few doctors would deny that it's a very real phenomenon. About 30% on average, in placebo groups respond, though for some reason in ME/CFS it's usually lower, at about 20%.
http://www.psychosomaticmedicine.org/content/67/2/301.short
>Samuel Wales:
poor lab controls, contamination and, of course, the ideological motivation that one is right.
How would you describe it?
Oh, you meant a patient taking a medication and responding well to it, instead of the lab quackery so elegantly on display of late. My bad. What part of "placebo" effect raised by Ed confused you?
Perhaps the data are, rather, explained not by the placebo effect so much as variety of the much-discussed homologous recombinaltion tiniker?
>From Gerwyn Morris:
VP=62 is a synthetic clone made from different prostate tissues which does not exist in nature
Lo Used a PCR assay which had failed to detect any kind of hgrv in lo atler
the other CDC and FDA assays used VP-62 as a positive control and thus were incapable of finding anything anyway
This left the NCI and wpi as the only game in town
the positive patients were all on meds that would have given a false neg
and the wpi were not sent the laboratory negative controls to validate as being negative in the first place
tests are not as straightforward as the marketing people would have you believe
for example it took 5 pcr runs until switzer was able to detect an amplicon in a patient known to be infected
Thus the level of concordance between the Ruscetti and mikovits serology on the Lo patients was very high
the design of the study was awful
they even outsourced the phlebotomy and the company in turn used inexperienced phlebotomists who performed all the coding
After the initial period HGRVs are cleated from the blood and become indetectable by PCR
The antibody response also fades away
The viruses remain readily detectable by PCR in tissue This is typical for a mulv class virus
RNA with sequence variability is intermittently detected in the plasma but of all assays only the wpi.s RT PCR could have detected this because their lower annealing temperatures and higher magnesium concentration will detect mismatches
>There is an abundant pubmed literature on the fact that protease inhibitors are directly immunomodulatory, effect apoptosis, affect t-cells, and are also directly anti parasite. And as mentioned above in comments, one ARV is effective against EBV, another against hepatitis.
Therefore, it is simply a hypothesis that ARVs offered a year reprieve from CLL because of a retrovirus.
All hypotheses must be entertained until there is solid proof and consensus.
And in reference to Jamie's previous post, there has to be more than, to paraphrase, I went off two of the ARVs because I didn't feel they were helping any longer, and I was feeling poorly, but I stayed on Viread…
This does not make much sense, as told.
Jill Neimark
>I just changed the blog settings to allow anonymous comments again, since it has been keeping people from joining in the discussion. I still encourage you to use your names, or at least a handle, so we can get an idea of who you are and consider your thoughts in context.
Btw, there are now over 3000 comments on this blog.
Jamie
>A simple question without hidden meaning provoked unintended
passion.
>Ed
VP-62 xmrv is a synthetic clone manufactured in Silvermans lab. It does not exist in nature.
Lo used an assay already proven not to work.
There was virtually no chance of WPI positives retesting positives because for some reason the patients were taking meds that would invoke false positives!
Once the initial viremic phase is over, gammaretroviruses leave the blood and are thereafter only found in tissues and the antibody response disappears.
The gammaretroviruses are only found in blood intermittently and are usually found as RNA when the virions enter the blood from other compartments.
Using PCR and serology assays optimised to detect a synthetic clone, which does not exist in nature, is never going to be able to detect a HGRV if present.
You dont have XMRV in your blood, you have a HGRV or possibly HGRVs.
Not all the negative controls had been validated as being negative because they had not all been tested.
>Ed,
I have outlined before, as posted in a previous blog why there is no other explanation for my response than treatment of a retrovirus. My screen for active infection with the different herpesviruses was negative. These ARV drugs do not have chemotherapy activity. Anti-telomerase activity has been proposed but anti-telomerase agents have had clinical trials and there are no reported response of human cancer in the clinic. Besides my response has been too rapid for depletion of telomeres causing apoptosis. Some of the retrovirologists have raised the possibility of selective toxicity which has never, ever been seen in Oncology. As for placebo effect lowering a leukemia count, I would discount this.
I believe that one of Dr.Coffin's virologists at Tufts has looked into endogenous retroviruses and CFS and has not reported any positive results. When one looks at what is happening here, a virus has caused proliferation of a B-cell clone and a T-cell clone. This fits the evolutionary strategy of a MLRV and has not been reported with a HERVK. The expansion of clonal gamma delta T-cells that you can see in my data may be a key issue as these cells are able to make the cytokines that are typically elevated in CFS and these cytokines have the ability to drive a simultaneous malignancy. This is called a paracrine activity and is documented. Clonal gamma delta T-cells actually may be the missing link explaining the known association of chronic inflammation and cancer. My results show that this may be happening and that the ARVs may be primarily working by lowering the amount of these clonal gamma delta T-cells. So your point that the ARVs are working via the immune system is what we are saying but specifically by targeting these clonal gamma delta T-cells.
We get to your final point of an unidentified retrovirus causing my disorder. I must remind you that three independent and excellent labs in the 1970s reported MLRVs associated with human cancer so these viruses have a history. One could consider other gamma retroviruses I suppose such as feline or primate, but there is no evidence for these being important and certainly this is not HIV or HTLV1 or 2 so we have run out of alternative unidentified flying retroviruses.
As to your lack of faith in Drs.Ruscetti and Mikovits, that is your opinion and I don't share it. Newer methods have become available as Dr.Mikovits points out. She would like to use them but as you must appreciate funds have dried up because Dr.Coffin says there is no infection in CFS.
In our survey we do see an excess of prostate cancer. Our survey has gone viral and the only built in bias is that people have to be able to read English and have a computer. The only requirement is that their disorder fulfils the Canadian Criteria for diagnosis of CFS.
From my patient population I have to share this. I have a physician friend ("AJ") who has 3 brothers, and all of the brothers had either CLL or prostate cancer. Dr AJ has recurrent prostate cancer and most recently developed CLL. He fulfils the Canadian Criteria for CFS. Dr.AJ's wife now has a very bad CLL and a very bad CFS. This is not so unusual in my experience.
Michael Snyderman, MD
>Dr Petrison,
Several things come to mind. First blaming a shortage of drugs on a ill patient population is a travesty. The drug companies have the ability to ramp up production but chose not to because it wouldn't be profitable. There is even a shortage of chemo drugs in the US and people who might be cured with chemo (there are some) are out of luck.
I agree totally that these drugs should be given in a rationally designed protocol so we could learn who would benefit and what drugs we should use. The problem is there are no funds for this especially now that we are supposed to believe that CFS is not due to a retroviral infection. It is not just the drugs that have to be paid for, we need to be able to follow some sort of parameters such as cytokines and NK function.
I did look for other viral infections in myself including all the herpesviruses except HHV8 which was not available and was unable to get any positive results.
I have not personally treated any one with ARVs except myself because of the medical legal issues. However I was the perfect patient to use them on because I have such easily obtainable parameters of disease and I didn't need approval of my peers to treat myself. I am glad I took ARVs and I believe they added at least a year to my survival. I am an Oncologist so I know my options and I also know my survival stats too. As the saying goes I was trying to make lemonade out of lemons and I believe my results can be used as a rational starting place for a drug trial. I am not yet done, I plan to substitute fosamprenavir for the raltegravir when I relapse again. By the way, I paid for my drugs out of my own funds.
Michael Snyderman
>I just knew from the moment I got sick that I had a retrovirus. What else could it be? I had all the classic symptoms Deep down in my heart and soul I knew it had to be a retrovirus, specifically XMRV. It was intuitive. I knew it was there. Even if my Ouija board and my XMRV test from the WPI hadn't confirmed this, I would still believe that a retrovirus is responsible for my illness.
Laurie B.
>Do we have any idea how many CFS patients are using ARV's? Dozens? Hundreds? i find it hard to believe more than that, but maybe I'm missing something.
Thanks, Lisa
>@Healers of disease
"VP-62 xmrv is a synthetic clone manufactured in Silvermans lab. It does not exist in nature."
VP-62 is a clone of the XMRV that Silverman initially isolated from prostate cancer samples, the same XMRV found in the 22RV1 cell line that was contaminating his lab.
"Lo used an assay already proven not to work."
So you are saying that Dr Lo deliberately decided to use an assay that he knew could not detect the virus, when he had one that could. If he changed his testing method that would have only been because he discovered that mouse DNA was contaminating his results.
"There was virtually no chance of WPI positives retesting positives because for some reason the patients were taking meds that would invoke false positives!"
Do you mean false negatives? So what about the ones that did test positive? And were the controls taking meds that invoked false positives?
"Once the initial viremic phase is over, gammaretroviruses leave the blood and are thereafter only found in tissues and the antibody response disappears."
In that case, how did the WPI ever find the virus in the first place, as all their blood samples would have been taken long after the acute infection? It takes at least 6 months to get a diagnosis of CFS.
"The gammaretroviruses are only found in blood intermittently and are usually found as RNA when the virions enter the blood from other compartments."
How come the WPI claim to have repeatedly isolated the virus from blood?
"Using PCR and serology assays optimised to detect a synthetic clone, which does not exist in nature, is never going to be able to detect a HGRV if present."
Well how on earth did the WPI manage it then? The whole thing was sparked by Silverman sending them VP-62 and them testing some of their samples against using PCR!
"You dont have XMRV in your blood, you have a HGRV or possibly HGRVs."
I thought it had left my blood to hide in my tissues? I can't keep up.
"Not all the negative controls had been validated as being negative because they had not all been tested."
The WPI agreed the negative controls along with everybody else. Anyway it doesn't really matter whether the positives or negatives had been tested, as the study clearly demonstrates that the WPI and Ruscetti's assays are about as reliable as tossing a coin!
>Sigh…
Even if Gerwyn's (AKA Healer of Disease's) arguments had any merit (they don't) the results still wouldn't make any sense at all.
After all, Mikovits and Ruscetti collectively designated 11 out of 15 "negative controls" as positive – and apparently "just" with assays calibrated to only VP62. Now, even if there was no pedigreeing done AT ALL regarding the controls, the odds of "accidentally" selecting 11 HGRV positive healthy controls out of 15 are astronomically low.
For instance, using Lombardi et al.'s estimate of 3.7% of healty people being infected, the probability of this freak occurence happening would be about 1 in 5 trillion (1/5,000,000,000,000). For your reference, the odds of winning the Powerball lotery are about 100,000 times higher.
>what part of my explanation do you not understand
>The Gamma retrovirus now discovered in 3 separate studies examining people with ME was given the wrong name.
The virus is a HGRV and not an XMRV.
HGRVs are known to cause severe neuroimmune disease in Apes and Cats.
Scientists also gave HIV the wrong name and initially all studies called in HTLV-III, when it was really a lentivirus.
>sorry my last post was directed at ed
In case anyone missed it
5 aza cytosine is a demethylation agent
methylation of cytosine is one of the epigenetic mechanisms involved in the transcription of genes
specifically methylation silences gene transcription
integrated GRV proviruses are heavily methylated.
This is how latency is maintained and in this state the grvs are not replicating and sont produce any virions or even proteins
to complicate matters further grvs integrate into areas of high guanine cytosine content(called CpG islands).The bonds between them are so strong that they can be very difficult if not impossible to amplify by PCR.Special conditions and reagent compositions are needed. PCR assaysoptimised to detect a free floating clone in a spiked blood sample simply wont cut it
As dr Mikovits demonstrated the addition of a demethylation reagent means that the grv is activated and starts replicating and hence rapidly builds in titre.This may well be the basis of PCR approaches of the future
The reason that I use the term grv is because the retrovirus detected in three separate studies investigating patients with ME was not an XMRV but a HGRV.Retrovirologists have got a nasty habit of giving a newly discovered retrovirus the wrong name.They did it with HIV and now they have done it again
>I understand all of it, but thanks for asking.
My argument is that, even if ALL of your arguments were true, your arguments would still fail to explain how:
At least 11 out of 15 controls "accidentally" turned out to be positive. I showed that the odds of that happening in real life are astronomically low, even if WPI did all of their pedigreeing wrong, and all of the positives were on drugs that made the HGRV in their blood disappear, and Lo used the wrong assays, and HGRV's do a bit of the old in-out with blood anyway…
In short, designating 11 out of 15 controls as "positive" for any (H)GRV (or whatever you want to call it), cannot be explained away by ANY of your arguments.
Now, what part of that do you fail to understand?
>@Ed
"VP-62 is a clone of the XMRV that Silverman initially isolated from prostate cancer samples, the same XMRV found in the 22RV1 cell line that was contaminating his lab. "
VP62 is a clone made from 3 prostate cancer patients. It does not exist in nature. For it to have found its way into 22Rv1 it must have contaminated that cell line post its creation in 2006.
"So you are saying that Dr Lo deliberately decided to use an assay that he knew could not detect the virus, when he had one that could. If he changed his testing method that would have only been because he discovered that mouse DNA was contaminating his results. "
HofD is correct. Two RT-PCR assays were employed in Lo et al. One found nothing the other detected 86% positive for HGRV. It was the negative assay Lo used in the working group.
I am not sure why you would suggest a method would detect MLVs? There is no reasoning to you leap. XMRV is not a mouse virus and the others have found HGRVs. Such a result would also have been reported in the paper. It is also possible that those leading the study directed them to do this.
"Do you mean false negatives? So what about the ones that did test positive? And were the controls taking meds that invoked false positives? "
False negatives will occur on such meds. There is much on the subject to be found in pubmed. That by itself destroys the validity the working group
Once you also understand that the paper makes it clear that not all controls were tested by all labs, there is no argument.
"In that case, how did the WPI ever find the virus in the first place, as all their blood samples would have been taken long after the acute infection? It takes at least 6 months to get a diagnosis of CFS " "How come the WPI claim to have repeatedly isolated the virus from blood? "
Although such retroviruses reside in tissue, virus does sneak out into the blood on occasion. To detect this highly specific conditions are required. PCR cannot begin to find low level virus in the blood without first lowering the annealing temperature and using RT-PCR. The right adjustments will pick up the virus. Again, pubmed has a wealth of papers on similar gammaretroviruses.
"Well how on earth did the WPI manage it then? The whole thing was sparked by Silverman sending them VP-62 and them testing some of their samples against using PCR! ""
The WPI used VP35. They hypothesised that if the virus was present it would be at a low level. Firstly because gammaretroviruses behave this way in relation to blood and tissue and secondly perhaps this was why other had failed to detect such a virus previously. They also used many of the specifics of Silverman RT-PCR assay, including use of VP35.
" I thought it had left my blood to hide in my tissues? I can't keep up. "
The virus favours tissue, but some will rarely sneak out into the blood.
"The WPI agreed the negative controls along with everybody else. Anyway it doesn't really matter whether the positives or negatives had been tested, as the study clearly demonstrates that the WPI and Ruscetti's assays are about as reliable as tossing a coin!"
The WPI, NCI and Lo labs did not receive all control samples for testing. The others were screened by with assays never shown capable of detecting anything other than a clone of VP62, that does not not exist. Of the labs that did screen certain controls they were unanimous, being in complete harmony or accord.
It is more comparable to say a coin was tossed by the working group, but the coin was never allowed to hit the ground. We have assays designed to detect a false clone that would never find a HGRV, together with patients on meds giving false negatives and controls never screened by those with the only validated assays. The working group failed in its purpose.
>@RRM
The controls found positive by the WPI, NCI and others were never pre-screened by those same labs with their valid assays. There is no longer any mention of the controls in this paper being pedigreed.
You can read both papers to see that Lo used a different assay. Similarly you can read about the affect those meds would have on positives. Why avoid these answers?
What are you failing to want to understand?
>@Muckle
I am not even arguing any of these points. Even if all your assertions were true, there is no chance in hell that 11 out of 15 controls "just happened to be" positive.
Although I think I am unable to convince you, but I'll rephrase, for the open-minded lurker:
Lombardi et al. proposed that 1 in 25 healthy people are infected. If that were true and you would randomly pick 15 people from the street, there would be like a 1 in 5 trillion chance you would accidentally pick 11 positive people.
Your (and or Gerwyn's) arguments do not relate to the validity of my calculation. "I just saw a tree so let's negate the results" would be about comparable.
>@RRM
You are jumping to conclusions. We have no idea where those people were from. They could all be positive. They were also not randomly picked.
Lombardi et al. found 4% positive. Lo found 7%. There is a paper circulating that is finding 14%. As techniques improve the numbers will increase, with some areas having a higher concentration of infected and others lower. HIV and HTLV are no different.
You are mistaken in thinking you have applied any calculation and your opinion is merely speculation.
>@Muckle
Ah, it's all becoming clear, the BWG deliberately picked controls that were positive and made all researchers other than the WPI & Ruscetti use assays that couldn't detect the virus. OMG they're evil! I hear they never blink. They're probably monitoring us right now. Let's switch to morse code:
–. . – / .- / –. .-. .. .–.
>@Ed
The other labs all used assays not diagnostically validated to detect a HGRV. All were only analytically validated to a clone that does not exist in nature.
>@Ed
Are you ok? You appear to be on edge?
>no RRM you think you understand it the fact is you dont
They did not preserve the PMBcs in Trizol or anything else.
The wpi assay alone depended on activated PMBCs( B and T cells) they need to be activated to increase in number so that the provirus integrated into their DNA also nicreases in concentration so that it can be detected
You cant activate or culture dead B and T cells without being treated with preservative such as trizol they would have been as dead as a doornail
the WPI was the only one in this part of the experiment which had PCR test known to be able to detect a grv in a clinically infected person
for some reason lo used a PCR test which had failed before ( he must be mortified)
the serology assays relied on culture so they would not have worked either what a mess!!
This is on top of people being on meds that would have probably led to false negative results any way and forgetting to send all the negatives to all the labs for negative validation
No wonder Lipkin was aghast when he saw this chaos
This is Gerwyn posting by the way
>The full picture regarding the Blood working group is emerging.
They failed to use Trizol or another preserving agent with the PBMCs. This would degrade those cells and prevent the WPIs assays from working.
>I have several areas to address and it may take a few posts (this is Jill Neimark, happy I don't have to register on Google just to post).
Dr. Snyderman, I assume you know Malcolm Brenner's work at Baylor? I do not know if you would be a candidate for his cell therapy, but when I wrote about gene and cell therapy I interviewed a man who had a B-cell blood cancer with a name that was somewhat long so I forget, had been bombed with chemo at Mayo and practically killed, was going to be sent home to die, had his son fly him to MD Anderson where they put him on a kindler gentler regimen and suggested he consult with Brenner. Anybody who has failed chemo is a candidate–assuming he's working on your type of cancer.
In this case, targetting EBV was the key to his sustained remission after only two injections of his altered t-cells. Basically, we are all infected with EBV, and eventually we make a seeming truce with it. It remains in a small percentage of our B cells, having adjusted its antigens to what are known as "weak" in terms of our own immune system response. Therefore our immune system recognizes the antigens (put up as flags on the cell surface so to speak), gets annoyed, but lets them live. "Grumbles, but moves on," so to speak. This is a problem when the cell turns malignant. Whether EBV can cause, or contribute to the cancer, is a big question. Certainly EBV is strongly associated with all kinds of morbidity including various cancers.
So basically the t-cells are removed, and are trained in the lab to target the weak EBV and kill any cell with the weak antigens as well. This fellow, in his 70's, had two injections of his altered t-cells, they killed off the cancer cells almost immediately, and he had been in remission for two years when I talked to him.
Not that many people know about this kind of therapy. People die for lack of knowing. I also spoke with lung and pancreatic cancer sufferers. So I think he deals with a wide range of cancers, and tailors treatments. It's been a few years since I spoke with him.
Just fyi. And if you already know about it and are not eligible, my apologies. This post may help somebody else.
On to my next point. I posted above that there is a robust literature on ARVs being immunomodulatory. Is everybody going to ignore this? They have been shown to modulate the response of t-cells. In addition we are starting to gather evidence they are effective in other viruses including the herpes family. This must be *strongly* considered as an alternate hypothesis to, "The ARVs helped me therefore I have an HGRV," end of story. A good scientist considers all hypotheses, and develops statistical methods for filtering out confounding data (in this case, you would have to find a way to prove that none of the other ways that ARVs work, or viruses they work on, are at all involved in your response).
Otherwise, you have a hypothesis. An interesting one, but in light of all the negative data this last year, far from proven.
In addition, I would like to say to everyone, that the fact something is a retrovirus is not special cause for shock and awe. A virus can do lots of damage without being a retrovirus. Consider Ebola, polio, etc. I don't mean this in an obvious way. Because HIV is in our consciousness, and because it targets the CD4 cell, the orchestrator of the immune response, we've come to think of a retrovirus reflexively as something extremely immune damaging. Post polio syndrome is apparently quite debilitating…and polio is not a retrovirus. And we already live with oncogenes and HERVs. Life is complex. I see in all this ballyhoo about the "retrovirus", a certain reflexive unthinking emotional response, that I think is related to the history of HIV in our country. But frankly, that thinking is not scientific.
On to my next post.
>The intensity of the attention on the emerging science is in danger of blocking the scientific process all together. We need to wait. The science is difficult and will take time.
Give it the time it needs and wait with an open mind.
We are so used to getting things immediately we have forgotten that not all areas of human endeavour are rapid. To understand something fully takes a long time. This was understood in the past but we seem to have forgotten it recently.
We do not know what is there yet. We cannot see it, it can only be indirectly detected.
Rapid judgements are bound to be superficial in this instance.
>This is Jill Neimark again. I posted this on Dr. Deckoff-Jones' FB.
I don't understand the change in her self treatment.
I don't think it's a trivial matter.
Why did she go off two of the ARVs? Having detailed the ups and downs of her recovery from bedridden to 80 on the Karnofsky, she says she did not post when she was feeling poorly (I would have liked to know what that meant, what symptoms returned) and that she went off two of the ARVs but not Viread, because…I'm paraphrasing, she had a feeling they weren't helping.
How is this decided? To seemingly randomly go on and off ARVs—–?
I also have no clue how, biologically, a retrovirus that could be producing what she has called a multgenerational epidemic, could be put into latency and kept there, as she suggests, after a short period of ARVs. Certainly HIV does not work this way. It rebounds in full force within weeks. Is there any sound theory for how a retrovirus that would bring down so many (theoretically) could then be put into latency quickly and remain in latency?
I certainly have no clue nor can I think of a biological means by which this would happen. But maybe somebody else does.
And if there is a retrovirus, isn't monotherapy dangerous?