German study finds xmrv

[leaves]

Hi IamME

I totally get where you're coming from. I think we have fallen victim to anxiety at your expense. I don't think you were doing anything that most of us weren't doing but, perhaps, you came along at a critical juncture for Leaves. It happens.

I'm sorry you were hurt, IamME. It's not easy to deal with such scary, unsettling and intimate things without having the occasional dust up.

I know how it is to feel really anxious, Leaves. This isn't easy, is it!

Peace out,
Koan

Well if my anxiety could make everyone feel better, lets go for that

 

[anciendaze]

auditory hallucinations and tinnitus

...Could I ask, how would you describe an auditory hallucinations? Sometimes, particularly just as I am drifting off to sleep (when I also experience tinnitus), I 'hear a bang or explosion' in my head [they're coming to take me away, ha, ha!]. My wife has them occasionally too, but they are a very strange experience.

Here's another example of a condition where preconceptions can make all the difference in how you are treated. Before anyone realized there were people without traumatic experiences with explosives who experienced this, you would have been a good candidate for a diagnosis of post traumatic stress syndrome. Since I did have such traumatic experiences, I'm glad I do not have to report such a symptom.

Hearing strange things, or seeing them, or even smelling them, while falling asleep, is actually a pretty common occurrence among healthy people. The general category is hypnagogic hallucinations. One common form is the feeling that someone is calling your name.

The general term for non-psychotic false auditory perceptions, presumably caused by damaged nerves, is tinnitus.

It is usually described as a ringing noise, but in some patients it takes the form of a high pitched whining, buzzing, hissing, screaming, humming, tinging or whistling sound, or as ticking, clicking, roaring, "crickets" or "tree frogs" or "locusts," tunes, songs, beeping, or even a pure steady tone like heard in a hearing test.[5] It has also been described as a "wooshing" sound, as of wind or waves.

People can hear music sometimes, or think there are voices too soft to be understood. This seems to be the brain trying to make sense of some random input from damaged nerves. Again, this can happen to people with no known psychiatric problem.

Healthy people can experience many hallucinations, without the influence of drugs or mental illness, simply from being sleep deprived long enough. Considering the wide range of sleep problems in CFS/ME, this could be expected.

More general auditory hallucinations do cross over into the realm of mental illness. If you read the different definitions carefully, I think you will see what I'm talking about in terms of overlap. Approach things with preconceived ideas, and results are predictable.

Even simply having hallucinations will not get you locked up, as long as you know they are hallucinations, and make an effort to separate private and shared reality. I think the serious trouble comes from acting on interpretations of perceptions not shared by others. This only differs in degree from a common problem of all human existence. We don't have a "mind link" to let us directly perceive what others do.

 

[ahimsa]

I'm looking forward to being a plague carrier.

That's quite a step up in social status for a CFS patient.

Thank you, Esther, for my first laugh out loud moment on the forums today.

I had another good one a while back when someone posted on another thread that "syndrome" really meant "Yeah, good luck with that." The people on these forums are just the best!

Okay, we now return you to the actual scientific discussion of XMRV studies. It's a bit too much over my head for me to join in but I'm enjoying reading the thoughts of all the others out there.

 

[serenity]

ha ha! i'm with ya, ahimsa! i love the funny parts, we need to laugh about it.

& it's so so true. i have lost one friend since i told my friends about XMRV. & that' fine, cuz i know - she's a jerk. nothing more to it than that. but it's been so unlcear all these years & with all the other friends i lost things were so much more complicated. now, it's simple. i have a disease, you choose not to stand by me - your'e a jerk. no discussion. so yeh, that's a big step up from the "it's really all your fault" game i've had to play in the past.

 

[Gerwyn]

I think when it is inserted in a place in the chromosome where it does not interfere with the proper working of the cell and is not activated it could just sitting there without giving symptoms.

Sorry lans i was not clear.it can be harmless if it stays there but retroviruses move around the genome either in the same cell or different cells.The likelyhood of it staying harmless is not high

 

[lansbergen]

Sorry lans i was not clear.it can be harmless if it stays there but retroviruses move around the genome either in the same cell or different cells.The likelyhood of it staying harmless is not high

Do they move ad random? Do they move without a triger?

 

[Gerwyn]

I read that a lot and i also know that some people (mostly in North America, i think) try to treat those infections. But then, why can't those infections be diagnosed? Or can they?
In my case, for example, doctors told me that they can see that i've had EBV, but from the type of antibodies that were found, they could also say that the infection must have occurred at least 6 months in the past. They never found an accute infection. Is this the case in others? Active viruses?
Because in this case that would be a criterion that allows to distinguish at least some PWCFS from healthy persons. I'm sure in this case there would have been studies that have found that in PWCFS there are more of those viruses present than in the average population. And this would have ended the "is CFS physical" discussion long time ago.

the problem is not an acute infection.EBV is another example of a latent virus.Detecting even a high titre latent virus is difficult.Detecting a gammaretrovirus is nigh impossible.These viruses do their damage when inserted into the DNA.they dont behave like other viruses.Because of the way they replicate they may not even produce antibodies

 

[Gerwyn]

Do they move ad random? Do they move without a triger?

yes lans I,m afraid that they do.It is called transposition.Pathogens can make the spread quicker but they will spread on their own

 

[Gerwyn]

I don't understand, Gerwyn. You say: "It could quite happily do nothing if not in a regulatory gene" but than you say "Will everyone with XMRV... have some health issues? Yes".
And besides, does people with HIV that don't have AIDS have health issues? If not - why do you think that everyone with XMRV will have health issues?

because XMRV does its damage when inserted which is different to HIV and inserts into start sites of genes.it moves around the genome.If it inserts into a regulatory gene which regualtes 40 other genes then some consequences are virtually inevitable

 

[lansbergen]

because XMRV does its damage when inserted which is different to HIV and inserts into start sites of genes.it moves around the genome.If it inserts into a regulatory gene which regualtes 40 other genes then some consequences are virtually inevitable

That would cause chaos.

You talked about inserting in start sites before. It is still not clear to me. Can you explain it again?

 

[Gerwyn]

Lol. You are driving me crazy . But i don't want to start a discussion again, so i won't talk about this on and on.

Some people were worrying about what will happen if the general public starts to become aware of XMRV and XMRV is actually dangerous. Since there is no certainty yet about which illnesses XMRV causes, the only way we can try to answer what will happen, is to make some thoughts based on assumptions. This is a CFS forum, so people here only have to be worried if XMRV and CFS are associated, because if that's not the case, most people here will not be XMRV positive.

I think the assumption of a prevalence of XMRV in the population of developped countries of 3% is a pretty good guess (we had the Science study, now this RKI study and the Japanese Red Cross values). If in healthy controls it's between 2 and 4%, then it will also be somewhere in that region (a bit higher of course, but probably not much) in the general poplulation. Or what percentage of the gerenal population do you think won't qualify as healthy controls (are not healthy)? Give them 100% XMRV positivity (which is too high as not every health problem is caused by XMRV or leads to XMRV positivity) and you can calculate how much higher than the 2 to 4% the prevalence in the general population can be at maximum.
Make a better guess, if you like, i'd like to hear. If it's higher than 3% that will mean less trouble for the XMRV positive people, because the more there are, the less possible it's to stigmatize them.

I also think the assumption of 0.3% of the population of developped countries having CFS is pretty safe. That would be ~1 million in the USA. Sometimes we hear even higher estimates.

I don't think you can say that 97% of the people diagnosed with CFS according to the CCC have XMRV.
From the WPI we have heard numbers as high as 99%, but you can't give an exact number since XMRV tests have not been done in large enough numbers around the world and are not reliable enough yet.
But for my calculation it does not matter anyway, because i was calculating for the (until now) hypothetical case, that XMRV is the cause of CFS. And in this case it will be 100% of XMRV positivity in true CFS cases, the others will have been misdiagnosed.

CFS has been around for how long? Around 25 years? So if it's caused by XMRV, which is probably the only case in which people here really have to worry about being percieved as dangerous, then, unless CFS develops a very long time after the XMRV infection, we can say that XMRV is not easily transmitted, because if it were, we would have tons of cases where one person in a couple or family has had CFS and later the other person/other family members got/have gotten ill too. Like in HIV, where it could clearly be seen that it's infectious, even though it's much less easy to transmit than influenza or other illnesses.

97 % of people diagnosed by the ccc crteria have the virus.That is published.The illness has been around for a long time possibly a hundred years or so .XMRV can well be described as a slow virus and could well take many years to produce symptoms.That is the whole point.The prevelance of XMRV in the general population is an order of magnitude greater than HIV.you know that making asumptions does not work in science.The only way to measure infectivity is to measure it. some Your figures are based on a 100% diagnosis .The actual figures quote that some 90% of people with ME are not diagnosed.Apply that to your equation and 97 % of me sufferers have xmrv.ties inquite well wih the published data.you are also forgetting that this virus could cause a number of different illnesses in different people.This is a large number of people.What is 4% of 310 million(the population of the usa) to me that is 12 million people not imillion.You are also forgetting that this is an oncovirus.I think people should be very scared

 

[Mark]

So I'm Mr insensitive now, am I? What about my feelings or is whomever gets in first? I feel Leaves has overreacted and left me feeling as if cast as some sort of board villain! I don't see why it should be my responsibility how someone else feels whether it's 10, 20 50, 80 or 99% of +XMRV people *could* go on to get ME/CFS or not. These are all possibilities, some likely and some no doubt extremeley unlikely -- and depending as I say on how a new CFS or XAND is defined as well -- if people don't want to consider these possibilities they shouldn't be reading scary research papers and discussing it on this thread, and certainly not trying to censor other people's ideas (even if they're silly!) As well as being responsible to each other we also have to be responsible to our own limits after all. Both my parents are dead and had multiple serious autoimmune diseases. We don't know what the exact significances of having XMRV are yet. I'm feeling quite hurt and taken aback about all this.

I'm really sorry if I unintentionally seemd to cast you as insensitive IamME. I intended to make a neutral plea to everyone to be careful and sensitive since all of us are facing some extremely tough issues here. I'm in the enforced single situation too actually, and I'd like to say it stinks don't it...all those taxes all going only one way and nowt for you or me...in fact I'll mention now that I've assumed I may well be infectious with something medically unknown and exremely nasty for roughly the last 10 years, and since I've not been up to much anyway I've kept myself in relative isolation for most of that time. I'm terribly sorry to hear what you say about your parents, that's a horrible thing to have to contemplate now, in the context of the emerging news.

Lest there be any doubt that I'm accusing you of insensitivity, I want to say very clearly that although I recall your sharp critique of one of my own previous posts, I thought your response was such a powerful statement, and as I remember, a fair and accurate critique, that I nicked your closing sentence for my sig because I felt you were somebody well worth listening to. I respect your bravery and honesty in confronting issues that may be difficult or painful, and your openness in sharing your experience.

I'm sorry if you feel your 'welcome' to Phoenix Forums has been less than warm. We are perhaps a little touchy of late, but I appreciate your presence here, and I'm sure we'll all stick together as we confront the difficult and painful possible implications of the latest research findings, and as we try to assess the most likely truth about XMRV and us.

These difficulties are presumably why convention would say that this is not a good thing for us to be doing, at such an early stage in the research, as patients - but I presume we are all here, writing and reading this thread, because we don't agree with that...

 

[Dr. Yes]

I think there are a couple facts and a bit of semantic confusion that need to be cleared up here...

they did use the same techniques Doc. there is lymphoid tissue in the lungs.T and B cells are lymphoid.The WPI looked in B and T cells.In old blo0d they would not have been replicating.That is the reason for the apparently complex methodology.The Germans went straight forthe lymphoid tissue in the lungs where the virus would have been replicating anyway(all mulv class viruses replicate in lymphoid tissue).They key however even at these relatively huge titres they still had to culture the sample to find XMRV by pcr!

Hi G,

I didn't find a part of the paper that mentions culture of these samples prior to PCR - I had looked, but all they mentioned was a parallel culture from the original samples to attempt to grow fungi or bacteria. I don't see an indication (yet) that they pre-cultured the samples on which they ran PCR. This is a very important point, so we should clear it up... if you know where they indicate that they did such a culture step, please let me know. If they did not, it suggests that the concentrations of XMRV may be much higher in respiratory mucosa than in blood.

The passage of the paper relevant to this issue, as far as I can see:

"All samples were analyzed by culture for pathogenic bacteria and fungi and by PCR for rhinoviruses, adenoviruses, enteroviruses, influenza viruses A and B, parainfluenza viruses 1–3, respiratory syncytial virus, cytomegalovirus, Epstein-Barr virus, and human metapneumovirus. All samples were tested in duplicates obtained by individual RNA extractions. XMRV RNA was reverse transcribed from total RNA, after which nested PCR or real-time PCR were conducted as recently described (1,12)."

In my first post I used the word 'techniques' broadly (maybe too broadly). When I said the German researchers did not use the same techniques as the WPI I meant that they did not follow the same protocol nor use the same approach (choice of tissue, no culture step that I can see) and possibly not the same primers (not sure about this) as the WPI. It was a different methodology (even if the principle was similar) as a result, and thus this cannot be called a replication in a strict sense of the term. However, it can be called a validation in the broad sense that it found XMRV in healthy controls (at around the same percentage, too) using a different methodology, and in some ways that is even better; it's especially a big deal in terms of the debate prompted by the three negative studies.

Of course the word 'techniques' could also mean specific types of techniques, like nested PCR, in which case the German study did indeed use at least one of the same techniques. (They also used others, that the WPI did not, on a small subset of patients, and still found XMRV).

The fact that respiratory secretions were used instead of blood is a major difference that should be highlighted; among many other things it (obviously) suggests a different place to look for XMRV.

The samples - from nasal and throat swabs, sputum, and bronchoalveolar lavage - would indeed contain lymphoid cells, but also epithelial cells (especially in the case of nasal or throat swabs). We should not assume that the lymphoid cells are the only ones that were infected with XMRV; it is important to find out which other types of (non-lymphoid) cells this virus may be infecting in humans.

Why do we think this paper used different methods to the WPI? I see thta one is looking at blood and the other at saliva but maybe the PCR is the same?

The original WPI science paper says:
"we isolated nucleic acids from PBMCs and assayed the samples for XMRV gag sequences by nested PCR (5, 6). Of the 101 CFS samples analyzed, 68 (67%) contained XMRV gag sequence"
If you look, reference no. 5 is to the Urisman paper that is also referenced in the German study under discussion in this thread. Reference no. 6 of the WPI paper is to their own supplementary materials that say "Nested RT-PCR for gag sequences was done as described (5)". Again in the supplement the Science article refers to, I assume, the PCR from the Urisman paper.

If you look up the original Urisman paper (free access - http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.0020025#ppat-0020025-sd002) and look at the supporting material download "Protocol S2. XMRV gag Nested RT-PCR", it describes a nested GAG PCR. Is this the one under discussion? Looks to me like this is the one used in the Fischer study we are discussing. I note also that Fischer is one of the authors listed the Urisman study.

Like others here this study looks like great news, but I am wondering if they have found it precisely because they used the same PCR test as the WPI? If so then it seems to me that this is not an issue of blood vs. saliva so much as demonstrating the difference between testing methods. If I have interpreted this correctly (I realise I might not have) then maybe this is more of a true replication study, in testing terms, than any of the others even though they were using saliva and not blood?

Hi Megan:

They did indeed use one of the same techniques, as I mentioned above, in that they used nested PCR on a gag sequence (not sure which one). However, this still could not be called a replication of the WPI technique for the reasons I also pointed out above; the samples and tissues used make a big difference scientifically speaking. If they did in fact culture prior to PCR, which I don't think they did, that would be closer to what might be called replication, but strictly speaking they would still have had to use blood for that term to be used (as well as closely followed the WPI protocols for preparing the samples, etc). You wondered if perhaps this study confirmed that you need to do nested PCR on gag sequences to find XMRV; however the WPI managed to find XMRV with PCR for env sequences as well. I think it is far more likely that the negative studies were a result of using insufficient concentration of virus (little or no amplification of virus pre-PCR), and a couple other methodological problems.
(Btw, this new study did not test saliva, but respiratory secretions from the nose, throat, trachea and lungs).

 

[bel canto]

IamME - there's a whole lot of scary stuff being discussed here. What you said was definitely not any worse than a lot of the other stuff. It must have just "hit" leaves in some way that you could not have predicted. Please don't feel like we think you did/wrote anything wrong. Your input is definitely valued.

leaves - I'm so sorry this hit you hard. If your mom is doing well, there's absolutely no reason to think that won't continue - nobody knows how one person is affected, and it's been pointed out that many people live very normal lifespans. And of course it may be that she does not have this retrovirus. It's very scary for all of us, though - I worry a lot about my family.

I'm so grateful to have this community of people to worry with - what an extraordinary group.

kind regards, bel

 

[Esther12]

re the heavy emphasis people are putting on culturing...

Does Mikovits mention it anywhere in her explanation about why the negative studies are not finding XMRV in CFS patients?

http://www.iacfsme.org/BULLETINSPRING2010/Spring2010MikovitsLetter/tabid/427/Default.aspx

I didn't see any mention of it, but I'm not really up on the terminology.

 

[jspotila]

97 % of people diagnosed by the ccc crteria have the virus.That is published.

Where was this published? My recollection is that WPI has made statements that they are finding 95% or more CFS patients positive for XMRV, but that this has not been published in any peer reviewed literature (yet).

 

[Dr. Yes]

re the heavy emphasis people are putting on culturing...

Does Mikovits mention it anywhere in her explanation about why the negative studies are not finding XMRV in CFS patients?

http://www.iacfsme.org/BULLETINSPRING2010/Spring2010MikovitsLetter/tabid/427/Default.aspx

I didn't see any mention of it, but I'm not really up on the terminology.

She doesn't address it directly there, but instead outlines the WPI methods that include the activation/stimulation/biological amplification/culture steps.

However, she and Ruscetti state it directly in their recent response in Science magazine (bolds mine):

Of the technologies used to identify and isolate XMRV in patients with CFS, PCR from DNA or cDNA from unstimulated peripheral blood mononuclear cells is the least sensitive method. We contend that the three recently published negative PCR studies (1315) do not qualify as being studies that fail to replicate our study, as neither the same PCR methodologies were used nor did these studies draw on the additional cell culture and immunological methods that we employed to observe XMRV nucleic acids and proteins.

 

[serenity]

i'm not sure i understand the touchy subject of parents, & i'm sure i don't understand any of the scientific stuff or even have the energy to try to read it - but to add my 2 cents... i'm an intuative type person, & my intuation always told me that my Grandmother who had Alzheimers had something in common with me. she also had a back injury, as i do, & her 6 children seem to always be angry that she didn't do enough while i have always understood that she was doing the best she could. she has passed, but we were very close. i will always remember her telling me "i thought pain was just a part of life." i have long suspected she & i had similar contidions. then, today while reading, a lightbulb went off - my dad has bi-polar disorder (i think many suspect i have inherited some of his traits tho i do not feel i am either depressed or manic) & my mom had colon cancer. both dad & mom are doing well, but it certainly makes me wonder...
or maybe i'm reading too much in. maybe it's too simple to assume one little virus could be responsible for so much. but i do gotta wonder???
what did i maybe get that was passed down thru my genes?

 

[anciendaze]

culturing/amplification

...I didn't see any mention of it, but I'm not really up on the terminology.

In that letter, she used the term "biological amplification", instead of the more common "culturing", probably because culturing has other meanings. The procedure included activating the cells, then allowing them to divide repeatedly, to boost the number of copies PCR would have to work with. She did not want people to think she was culturing virus using a cloned cell line which might be contaminated.

 

[Esther12]

Thanks for the responses. Off to bed now, but I'll have a good read tomorrow.