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Transcription of Judy Mikovits Prohealth Lecture

Discussion in 'Media, Interviews, Blogs, Talks, Events about XMRV' started by thefreeprisoner, Jan 23, 2010.

  1. froufox

    froufox Senior Member

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    There are still some things she said in the bit that I transcribed that I cant understand - I have highlighted them in black. One is about the techniques the other studies used which is obviously important to know, but I've listened lots of times now to no avail. If anyone else can work out what she is saying could you post a msg and I can edit my post. That would be great thanks.
     
  2. V99

    V99 *****

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    froufox, I think I have worked out what two of you missing bits are.

    e, satisfied folks postulates as we know them for viruses - for an adult T-cell leukaemia, and this is a very aggressive leukaemia and the mechanisms for how it causes that are still largely unknown. But at any rate 10-20 million people are infected, but you see very few - only sporadic cases occur in the US or Europe and the US incidence is only about 0.2%. They dont even test for it in the blood supply because its just simply not a problem in America, its endemic in the regions that are shown here today.

    And the second argument that supports maybe whats different between these studies is the transmission from the actived PBMCs .. so if I take the white blood cells, some of which where I can't see virus and just put them on LNCaP, I can transmit the virus to this indicator cell-line that has shown you because its defective in RNaseL (theoretically because its defective in those, but we learn more about it later), will amplify and replicate high levels of the virus. So there are scientific reasons why there are differences between these studies, but I dont think there is any doubt that XMRV is a new human retrovirus, and since both HIV and HTLV1 are associated with neurological diseases and cancer, and now we have associated them with a neurological disease and cancer, that this a real human pathogen.

    So recent publications after those publications (I'm just walking through the literature off the last few years) might give us a clue to the pathogenesis - how XMRV might cause disease. So this paper by Steve Goff's lab shows that XMRV establishes in an efficient infection, and spreading infection, thats enhanced by transcriptional activity in prostate cancer cells. And what that means is, I told you the receptor is on every cell of the body, but clearly every cell doesnt have the machinery necessary to replicate the virus to high levels. In fact we see that the peripheral blood mononuclear cells really don't, and thats why we dont know where the tissue reservoir is. So he simply infected a lot of different cell-lines and he found that the expression was very very low level except in essentially one cell-line and thats LNCaP. So we got very very lucky in that this was the only cell-line I thought about as an indicator cell-line....we could have screened the hundreds of cell lines I know of that we do regularly when we're looking for viruses because if you can't grow it you can't study it.

    So LNCaP turned out to be really serendipitous and I think the key technical advance to being able to make that discovery, its just clearly luck. He showed that LNCaP responds to androgens, I told you it lacks interferon and RNA cell anti-viral responses, and I'll show you whats called the promotor and describe , the response elements, that might give us a clue as to the pathogenesis. And then Bob Silverman's lab showed the same thing, he showed that


    What do you think?
     
  3. kurt

    kurt Senior Member

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    Thanks to all the transcribers. This is great work (and so fast!).
     
  4. garcia

    garcia Aristocrat Extraordinaire

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    This is actually a tough one to work out unless you know the term. She actually says:

    "satisfied Koch's postulates as we know them for viruses"

    More on what Koch's postulates are here:
    http://en.wikipedia.org/wiki/Koch's_postulates

    I think she says there:

    This is a mouse b-cell line that expresses the erythropoietin receptor
     
  5. Lily

    Lily *Believe*

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    Hi froufox -

    Here's what I think:

    In paragraph 2, the word you are looking for is erythropoeitin

    In paragraph 7, It's Norbert Bannert (instead of Norman)

    In paragraph 8 where you have ???, she actually didn't finish her thought there, and I think she mis-spoke and then went on with what she intended to say instead - she sort of corrected herself -

    In paragraph 9 where you have satisfied folks - I believe that should be Koch's postulates

    In paragraph 12 promotor, is actually promoter


    You did a great job!!!! Thank you soooooo much!!!!:D

    ETA: Wiki says: Koch's postulates are four criteria designed to establish a causal relationship between a causative microbe and a disease. The postulates were formulated by Robert Koch and Friedrich Loeffler in 1884 and refined and published by Koch in 1890. Koch applied the postulates to establish the etiology of anthrax and tuberculosis, but they have been generalized to other diseases.

    The postulates are:
    The microorganism must be found in abundance in all organisms suffering from the disease, but should not be found in healthy animals.
    The microorganism must be isolated from a diseased organism and grown in pure culture.
    The cultured microorganism should cause disease when introduced into a healthy organism.
    The microorganism must be reisolated from the inoculated, diseased experimental host and identified as being identical to the original specific causative agent.
    http://en.wikipedia.org/wiki/Koch's_postulates
     
  6. froufox

    froufox Senior Member

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    Thanks guys!! That was a big help, I've corrected it all now. Well done on getting Koch's postulates, I'd heard of Koch but not heard of Koch's postulates! Thanks for posting the info about the postulates Lily thats very interesting.

    It took me a while to transcribe as Judy M talks so fast but it was fascinating to do at the same time even though it was quite technical - a good learning process!

    Thanx again.
     
  7. Advocate

    Advocate Senior Member

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    I'm going to try Section 7, right now. I say try because I was unable to get normal sound on my computer. So I taped it into an old recorder off another computer, and again, the tape recorder adds another layer of bad sound. If I can't do it, I'll let you know.
     
  8. Advocate

    Advocate Senior Member

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    Nope, it didn't work. Maybe I'll try another computer tomorrow.

    The first time I watched the Mikovits video the sound was fine. Today it is unintelligible.
     
  9. Lily

    Lily *Believe*

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    I'll do video #2 Section 8, 30 mins-45 mins.
     
  10. JAS

    JAS

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    I wil do Video No. 2....45 mins - end. I do have a problem as I cannot unfortunately hear the questions, would it be ok if I just transcribe what Dr Mikovits says but leave the questions out?
     
  11. fds66

    fds66 Senior Member

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    Been tied up all week so not been able to transcribe. I think that all is covered now but difficult to tell. If there is anything left to do then let me know and I'll do some. If anyone is having problems and needs help let me know. Well done everyone.
     
  12. fds66

    fds66 Senior Member

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    One other thing - where is the completely transcript going to be?
     
  13. garcia

    garcia Aristocrat Extraordinaire

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    Presumably Cort will put it up like he did with the other transcriptions. But this time I would like the slides (since we have access to them) interspersed with the text as pictures if possible? That way you have the text & the slides all in the same place for convenience.
     
  14. Lily

    Lily *Believe*

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    Dr. Mikovits transcription

    If anyone sees an error in my interpretation of what I thought I heard, or some incorrect terminology, I'd appreciate it if you would let me know. Thanks in advance;)

    Dr. Judy Mikovits, January 22, 2010, Video 2 Section 8, 30 mins-45 mins

    (Q & A)

    Question: Inaudible

    Dr. Judy: We have looked in a limited number of families that we’ve done, and in fact, maybe only one member of the family has CFS, and so it will be an interesting year, but, um, once these families and the various diseases and trying to understand – it’s a very slow replicating virus so it really just sits there for a long time.

    So if you can keep the reservoirs low, you might have the virus your whole life and never get sick. We don’t know how long it’s been in the population. We think like with other retroviruses, the younger you get sick, the more severe the disease.

    If you give it to the rats when they’re neonates, they get cancers. If you get it at 30 or 40, they don’t get anything.

    So the immune system is educated and grows as you go along. It may be more fragile at different times in your life. We’ve seen a lot of it at puberty, boys and girls alike. There’s a lot of infection or at least apparent infection, disease occurrences at 12 or 13. And I do know that’s when Andrea got sick. (to Annette Whittemore) And presumably, your family was in the same space, but there was no other disease or child you had at puberty at that point.

    Retroviruses don’t infect people differently. You can’t go to Germany and say the reason they don’t have it is because they have hardier genes in Germany, because everybody gets infected. It’s just which immune system can control the virus and keep it down.

    Since we’ve been able to treat HIV/AIDS now, we find there are elite controllers. People are walking around with HIV who never knew they had HIV, and the copy numbers are all low. Their immune system is fine, and they have no idea when they got infected.

    Question: So a lot of patients say they had a flu, a weird flu, a week or two before they got CFS. So what you’re saying is that flu-like illness is a bug that came along and allowed the XMRV to create CFS?

    Judy: Well, no. What I think is happening is – I know almost nothing about CFS – but what we think is happening is – remember the slide where I showed you those little events? You know, what was the event that was the straw that broke the camel’s back?

    Where did the balance tip between, where you’ve got your immune system working well and the virus and the immune system are co-existing just fine, then some other, that other bug, whether it be lime, a flu, a anything, GETS YOU.

    And the virus, the cells divide, and cause the B and T cells you need to mount an immune response. And now you’ve got your memory population that might have been harboring the virus and it’s replicating ‘cause it’s seen that same pathogen before, so it could be a common everyday pathogen, and then you just tip the scale to where now your immune system can’t handle it or any. And every day you’re seeing more and more infection ‘cause you’re NK cells aren’t working, your B cells aren’t working.

    We put that antibody up there for a reason, we haven’t been able to correlate the levels yet, because we haven’t been able to find high enough numbers, but we do see these infected families where infected spouses and things have very high levels of antibodies, that suggest maybe these antibodies in the retrovirus can be protective, and maybe there’s an immune therapy on the horizon as well.

    So you can think about it in that way. It doesn’t have to be the insult. You might not know how long you harbored that virus, or it could be….

    Question: So what you’re saying is XMRV was there, then something else came along that was the tipping point.

    Judy: Yes, that was the tipping point, that’s correct. That’s our hypothesis. And again, it’s testable, we don’t know.

    Question: (partially inaudible) Is there differences between – ‘cause you’ve got a big population with sudden onset and then you’ve got a big population that had a gradual onset. And a lot of the gradual onset patients are worried that they don’t fit this equation. So what would you say to that?

    Judy: That the bump is smaller, it’s not a huge burst. So little insults over time. I think, for me anyway, I know it’s only a handful that I’ve looked at, that I know the patients and I know what the onset was. So sudden onset, gradual onset – it just depends on what the environment was, the triggers and what the events were that spurred it on. So I don’t see that if you were gradual onset, that you might not be infected as well.

    Question: That’s an important thing to know.

    Question: Have you tracked any of the inflammatory markers in the blood with the XMRV virus?

    Judy: Yes.

    Question: What are you finding?

    Judy: We find a signature suggesting a viral infection, an unclear viral infection. So we have 5-10 inflammatory cytokines and chemokines that will cluster in an infected person. But the problem is, that we don’t know if it indicates active infection. Certainly when the virus is quiet those cytokines will go down and those cytokines will change, and it might be a nice biomarker for following active infection, but we haven’t analyzed the data in that way yet. So it could very well sit, and some of those go down and up very quickly. So like 3 days you’re up at IL-6 or IL-8, then some of the chemokines are elevated and then they go back , like an EEG. Just like you might assume the retrovirus could be an EEG, depending where in the body that is is. We don’t know the reservoirs, so we don’t know what’s controlling it.

    Question: I’m confused about something. You say you can’t find it very easily by PCR, so you are culturing it. Then how did you find it for the Science paper?

    Judy: Well, because I found it in 67%, by PCR, of the patients, but I looked several times, at any given time. I just said it's like an EEG. So I got lucky and found it when the patient was high. They’ll come to the doctor when they’re sick which might mean they’re replicating more WBCs and there’s more virus in their WBCs.

    Question: So you checked these patients multiple times over time?

    Judy: No, I did them all at the same time because they were in the repository. So we collected those samples multiple times over time.

    Question: So you took like 10 samples from one patient?

    Judy: No, usually it was four. A Poisson distribution, the copy number could be as low as 5 or 10 copies per mil of blood, so there’s a statistic called Poisson distribution, you might find it one out of three times so we went more than one and in most people we found it.

    Question: So the UK paper had studied patients. If they would have looked at 4 or 5 per patient, do you think they might have found it?

    Judy: They might have certainly found more, yeah.

    Question: They essentially accused you of contamination.

    Judy: Right. But why would I have contaminant in my sick people and not my healthy people? How would I do that? I did it in three different labs. I did it in Cleveland Clinic. When I first came to the Institute, we didn’t have a lab, so everything we drew from around the world, I sent to Frank’s office. They processed it. I went there, I put it in the microarray or whatever I was going to do, and we worked there for a couple of weeks, you know, while we were building our lab and finally got started.

    So there, from the way we did the study there’s just NO possible way there was contamination. And that’s what the reviewer concluded. And the phylogenetic analysis proved two things. It wasn’t a mouse virus, it’s a human virus; it wasn’t a contaminant from a lab. We never do mouse work anyway, but it wasn’t a contaminant from mouse feces or something in the lab and it was clearly a new branch of the (phylogenetic) tree. A human virus. And our virus wasn’t exactly the same as the prostate virus. It’s still XMRV, because it’s 99% the similar. But that’s enough to show it wasn’t a contaminant.

    One of the things you have to use to get a good PCR is at least 750 nanograms of DNA. They have no idea how much DNA was there. And they quantified 3-9 out of 186. Sure they found a band of globins, but globins are in every single cell, so again, you’re making an unfair comparison of what you’re saying you see. And then you amplify it for less cycles than what would really push the envelope.

    Also to show no mouse contamination with the CDC, and Bill Switzer - after he saw the results in the paper. He said, “I have an assay that’ll show it’s a mouse virus contaminant. It’s a very sensitive, very specific PCR. Will you do it?” I said, “Sure, send it to me”. And we did it on all 100 and not a one. Not one cell line in our lab. He’s found a couple in his lab, but we didn’t find any. Perfectly controlled. He said, “Congratulations, it’s not a mouse contaminant.”

    So there’s little else we can do except wait for the rest of the community. It’s there.

    And the prostate people say, “oh you didn’t find it in 500 people, it must not have anything to do with prostate cancer. It just didn’t have anything to do with that population.”

    And again, Norbert Bannert, is a high quality scientist and as soon as he saw the paper he called me and asked for the reagent. Because he’s gonna go back and look to see if it really is there, and help us find some answers. He’s also looking to see a CFS group. So it just depends on what you really want to find. We weren’t biased in our study. You know, I’m a cancer cell biologist and we aren’t biased.

    I had to work really hard to get most of the people – not Frank and company. The NCI didn't know what CFS was. You know, fortunately, our scientists can at some level do what they want as long as it’s along with the mission. I remember one gentleman, high level NCI official, said “tell ‘em to get over it”. You know if you can’t, and again that’s a credit to Annette and the formation of the WPI. Um, we got a grant where we literally named XMRV within my first six months in 2007, because of the juxtaposition of seeing that paper in prostate cancer, right when we met.

    And thinking about the possible mechanisms and the grant got rejected three times, you know, because scientifically, retroviruses aren’t in CFS. If you would have looked in Wikipedia in August, it would say retroviruses aren’t in CFS. It doesn’t say that anymore. So we made progress.

    So will there be a variant? Maybe England will be a variant. Maybe there’s XMRV2, and one causes more. Maybe we found the one that causes the least severe disease, so maybe there’s XMRV2.

    A group emailed me from China, and the guy said “please”, you know in very broken English, “please don’t leave me, please write me back. Please help me”. And so Sam Chow was going over there, and I said will you look up this group? And Sam Chow has found a virus that looks similar. Might not necessarily be the same virus, might be a lot more aggressive virus over there. Now that’s just anecdotal.

    Question: Inaudible (something about SARS?).

    Judy: I don’t know, I’m not familiar with that literature.

    So we don’t know. The good news is we have something to work with that is a very testable hypothesis. This has been very rigorous. You don’t get more rigorous than Science and Frank and Sandy Ruscetti. They’re highly regarded. That entire team are experts. So the CFS population had the opportunity to have them look at us and they didn’t show any bias. They looked. You know, I asked Frank to come out to Reno before I took the job, because I said, you know, I hear a lot of things about this. So he came out and stayed a few days and saw some patients and spent some time with them. And I was on an East coast trip working. And I came back a few days later and I said, “Well, what do I do?” And he said, “Take the damn job!”.

    So if you’re really looking unbiased and you look at this, it COULD be only 10,000,000 in America, it could be only endemic right here, it could be like Japan. And HTLV is fairly innocuous – only 20% get that tropical spastic paresis.
     
  15. fds66

    fds66 Senior Member

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    Thanks Lily,

    The statistics that she was talking about with low copy numbers is called poisson statistics. You've written SANS in your script I think. It a mathematical way of predicting probabilities.

    Hope that helps

    Corrected:


    Judy: No, usually it was four. A Poisson distribution, the copy number could be as low as 5 or 10 copies per mil of blood, so there’s a statistic called Poisson distribution, you might find it one out of three times so we went more than one and in most people we found it.
     
  16. Lily

    Lily *Believe*

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    YAY! thanks so much fds66!!!!!
     
  17. garcia

    garcia Aristocrat Extraordinaire

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    Judy Mikovits says:

    Well because I found it in 67% by PCR of the patients but I looked several times, at any given time. I just said it's like an EEG. So I got lucky and found it at a time when the patient was high ...

    The missing word is "know".

    I remember one gentleman, high level NCI official, said tell em to get over it. You know if you cant, and again thats a credit to Annette and the formation of the WPI. We got a grant where we literally named XMRV within my first six months in 2007, because of the juxtaposition of seeing that paper in prostate cancer, right when we met.
     
  18. Lily

    Lily *Believe*

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    thank you, garcia!!!!
     
  19. JAS

    JAS

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    I have transcribed Video 2, 45 mins to the end, there are only a few little wrinkles! Just want to check it through tomorrow when I am more awake and then will post.
     
  20. Wow JAS - all of it? Right to the end?
     

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