#109~ The Birth of "Advocates 4 Answers"

OH Happy Day ♥ Happy Birthday Dr Judy  

and for the birth of "Advocates 4 Answers"

Happy April 1st everyone... and it's NO Fools Joke !

Please Help us wish our very own, Dr Judy Mikovits' Birthday today !!  

Birthday Greetings  for Dr. Judy maybe emailed HERE

Also, Congratulations to Dr. Judy in her New Position at WPI as the 
Director of Translational Research. What a perfect fit for her and blessing for WPI and the patients.

What a Perfect Day to Launch the New Website and New Advocacy Campaign  
for Neuro-Immune Diseases. 

Dr. Judy and WPI epitomize the essence of real

science, passion and compassion~ along with now a new advocacy campaign ~

 The website is UP and Available NOW for you to Sign the Guestbook, so you can Help be one of the "Advocates 4 Answers" and becoming a part of WPI's "May Awareness Month" for neuro-immune disease.

 

Throughout the month of May, we are inviting you to join us in raising awareness and funds for the important research into the underlying causes of neuro-immune diseases. Every day there will be an opportunity to show your support. By signing our guest book, you are pledging that you want to be a part of our movement and will participate at a level you are comfortable with to show your support.

You will receive an email prior to May 1 to help get you started. 

You will also be show the link for the New Facebook page link for this Campaign also...

Be an "Advocate 4 Answers" and sign up today~ by going here ~

Please SHARE this post and  get everyone you know 
friends, family, and care-takers to help and participate. 

http://anida.com

We look forward to an Exciting 2 months of Advocacy and Beyond......

Mobilize and "Join Our Energies TOGETHER" so we can be Positive Source for advocacy at this much needed time with all of the recent research developments thanks to WPI and our Heroes "on the Reno Hill."

I sincerely look forward to Joining with you ALL and taking part in this 

"much needed" long over-due United Campaign for the millions that these illnesses effects.. 

Its it TIME to Unite behind the diseases and drop any political illusions of us being separate and work with the one place that we already KNOW and TRUST ♥

We are as Powerful as our United Voices.

Our time HAS come.. Let's Grab this Brass Ring and Run with it, OK ?

We are now on our 3rd Generation that is being effected with these illnesses and it is Time NOW for us since the Universe has seen fit to unite us with this technology to help Be "Our Own Voice" ~ 

Please Come Join us... 30 years is long ENOUGH

It is TIME for the People to USE their Voices..



XMRV will NOT Quit and Neither will WE or WPI, OK

Heartfelt Thanks for Participating and Joining us 

on this New Campaign of Advocacy !

ShareThis

#64~ Letter to the Editor by Dr Judy Mikovits~XMRV+ U.K.

Letter to the Editor

---

Judy A. Mikovits, PhD, Director of Research, Whittemore Peterson Institute, Reno, Nevada

    Since the publication of the Science article in October 2009 concerning Xenotropic murine leukemia-related virus (XMRV) in patients with chronic fatigue syndrome (CFS) (1), there have been three PCR-only papers published that have been unable to detect this agent (2-4). As these papers have generated questions concerning the presence of XMRV in persons with CFS, we wish to document the precise methods used when XMRV was amplified from the patient samples. We would urge researchers not to rely solely on DNA PCR on unactivated PBMCs, and offer our assistance if needed.

Why are reports of negative PCR studies occurring?

    The first possibility is that like HTLV-1, and unlike HIV-1, the worldwide distribution of XMRV is low, particularly in Europe, and that chronic disease such as CFS may have varied environmental triggers in different parts of the world. In our subsequent work since the publication of the Science paper, we have found XMRV in persons from around the world and we think this explanation is unlikely.

    A second possibility is that there is more sequence diversity in XMRV than previously reported. That is, because of strain differences, PCR detection will occur only if the primer sequences employed are correct for that particular strain. However, PCR sequences generally are from conserved regions of the viral genome, so this explanation may also be unlikely.

    The replication rate for XMRV could be very low and/or the levels could fluctuate in a given individual over time. This explanation may apply for individuals, but would be unlikely to apply to large numbers as the three negative studies have used. However, if any of the last three are true, single round PCR of genomic DNA isolated from PBMCs, using primers based on published sequences from using highly specific PCR based on the sequence of a single molecular clone, (VP62) might not result in the amplification of XMRV, even from an infected individual.

    A fourth explanation is that peripheral blood mononuclear cells might not be the main reservoir for the virus. It should be noted that early in HIV infection, PCR of peripheral blood mononuclear cells will not detect the virus, as the reservoir is in macrophages. It is only later that CD4 cells become infected and PCR of peripheral blood becomes positive. From in vitro studies we are aware of tropism of XMRV, however no full investigation of in vivo tropism has been carried out to date. There are other possible explanations for the disparity of results between the Lombardi et al study and the PCR-only studies which are currently being explored.

    However, despite the current assumption that PCR answers all questions, there are serious flaws in this assumption, and PCR-only papers are rarely published without supporting data. For this reason, the Science paper employed several other biologic technologies in asserting that XMRV is an infectious retrovirus, and a human pathogen linked to persons with CFS. It should be noted that the question of XMRV "causing" CFS was never discussed in the Science paper.

    Because of these issues, we will describe below the methods we and other investigators currently use to reproducibly detect XMRV. Because we wish to accurately share technical information, the following will be quite technical, and possibly difficult for clinicians. Again we would like to offer any assistance we might be able to give in discussing these methods.


AMPLIFICATION METHODS FOR XMRV DETECTION:

1) REVERSE TRANSCRIPTION (RT)-[PCR]

    To avoid problems with laboratory DNA contamination, we perform nested PCR with separate reagents in a separate laboratory room designated free of high copy amplicon or plasmid DNA. Negative controls (absence of added cDNA or DNA) were included in every experiment. Two methods are used. TRIzol method is used to extract RNA. PBMC (2-5 million) or plasma (250 μL) are added to 1 mL TRIzol reagent (or 750 uL TRIzol LS reagent) and RNA prepared according to manufacturer’s instructions. The final RNA pellet was suspended in 14 to 30 μL of RNase-free water, and quantified using the Quant-iT RiboGreen RNA (Invitrogen). Reverse transcription (RT) with SuperScript VILO cDNA synthesis kit (Invitrogen), cDNA was made by a mixture of oligo d(T) and random primer with 2.5 μg total RNA. Subsequent optimization of the nested PCR revealed that an increase in starting template leads to a reduction in false negatives. 

    Identification of XMRV gag and env genes was performed by PCR in separate reactions. Reactions were performed as follows: 750 to 1000 ng DNA/cDNA, 2 μL of 25 mM MgCl2, 25 μL of HotStart-IT FideliTaq Master Mix (USB Corporation), 0.75 μL of each of 20 μM forward and reverse primers in reaction volumes of 50 μL. For identification of gag, 419F (5’-ATCAGTTAACCTACCCGAGTCGGAC-3’) and 1154R (5’- GCCGCCTCTTCTTCATTGTTCTC-3’) were forward and reverse primers. For env, 5922F (5’- GCTAATGCTACCTCCCTCCTGG-3’) and 6273R (5’- GGAGCCCACTGAGGAATC AAAACAGG-3’) were used. For both gag and env PCR, 94 °C for 4 min initial denaturation was performed for every reaction followed by 94°C for 30 seconds, 57°C for 30 seconds and 72 °C for 1 minute. 35 cycles was performed followed by final extension at 72°C for 2 min. For second round PCR, annealing was performed at 54°C for 35 cycles. Six μL of each reaction product was loaded onto 2% agarose gels in TBE buffer with 1 kb+ DNA ladder as markers. PCR products were purified using Wizard SV Gel and PCR Clean-Up kit (Promega) and sequenced.

    Alternatively, in the first round of PCR, cDNA is synthesized followed immediately by the first round of PCR amplification using USB Taq polymerase and master mix according to the manufacturer’s instructions, using 0.5 ug RNA as template and the previously described GAG-O-F (F542) and GAG-O-R (R1153) as the sense and antisense primers, respectively (1). The second round of PCR was performed using 5 ul of the reaction mix, the primers GAG-I-F (F603) and GAG-I-R (R1015) using 0.5 ul USB Taq polymerase and Invitrogen PCR buffer, supplemented with an additional 0.5mM MgCl2. The PCR products were then separated on a 1% agarose gel, and the PCR products of the expected size (413 bp) were recovered with a QIAEX II gel extraction kit (Qiagen) and sequenced. Using a similar approach for the amplification of env sequences using previously described primers only resulted in the amplification of a small percentage(gag positive samples.


2-VIRAL AMPLIFICATION BY TRANSMISSION TO LNCaP (Biological amplification)

    Two methods are used to detect virus following transmission to LNCaP cells. The first uses nested PCR: Plasma from 20 mL of anti-coagulant blood is flash frozen and PBMC isolated by ficoll-hypaque density centrifugation. The PBMC are activated for three days RPMI-1640 medium supplemented with 10% fetal calf serum, 2 mM glutamine, 1 mM sodium pyruvate and antibiotics supplemented with PHA (1 μg/mL) and IL-2 (20 units/mL). In 15 mL centrifuge tubes, 5 x 105 detached LNCaP, 1 x 105 activated PBMC free of IL-2 and 50-250 μL of autologous plasma in 250 μL of RPMI complete media are centrifuged for 10 min at 1500 rpm. The entire contents of the cell pellet are cultured in a T-25 flask in complete RPMI 1640 media for 4-5 days until the LNCaP cells are confluent. DNA is then extracted from the cells and nested PCR for gag is performed as previously described. A companion negative normal donor is always run under the same conditions.
  
3-VIRAL ISOLATION

    Cell-free plasma (100-150 μL diluted to 300 μL with serum-free tissue culture media) is added to a six-well culture plate with the LNCaP cell line (30-50% confluent). Negative normal plasma is included in one well in each plate as a control. The plates are centrifuged 5 min at 1500 RPM, rotated 180o to prevent drying out part of the culture and centrifuged again for 5 min. It is important to minimize toxicity in the plate and conditions will vary with type of centrifuge used. Each well is given 3-4 mL of complete RPMI. For cell-cell transmission, 1 x 106 PBMC given PHA but no IL-2 are added to a six-well culture plate with the LNCaP cell line (30-50% confluent) are centrifuged and cultured as above. For PCR determination the cell/plasma samples are culture for 5 days then harvested. For viral isolation, after overnight incubation, the plasma/PBMC is removed and fresh media is added. The cultures are grown until confluent, expanded by transferring to T-75 flask. Productive virus isolation is determined at passage 2 by immuno-blotting or PCR detection and sequencing.

        In sum, we have had success in identifying XMRV with the above methods. We remain confident that XMRV is linked to CFS and further studies are proceeding. We are aware that the research concerning XMRV's role in persons with CFS and other neuro-immune illnesses is in its infancy, and eagerly look forward to emerging knowledge in this field.



1.    Lombardi V, Ruscetti F, Gupta J, Pfost M, Hagen K, Peterson D, et al. Detection of an infectious retrovirus, XMRV, in blood cells of patients with chronic fatigue syndrome. Science. 2009;326(5952):595-89.
2.    Erlwein O, Kaye S, McClure M, Weber J, Wills G, Collier D, et al. Failure to detect the novel XMRV in chronic fatigue syndrome. PLoS ONE. 2010;5: e8519. doi:10.1371/journal.pone.0008519.
3.    Groom H, Boucherit V, Makinson K, Randal E, Baptista S, Hagen S, et al. Absence of xenotropic murine leukaemia virus-related virus in UK patients with chronic fatigue syndrome. Retrovirology. 2010;7(10 [Epub ahead of print]).
4.    Kuppeveld F, de Jong A, Lanke K, Verhaegh G, Melchers W, Swanink C, et al. Prevalence of xenotropic murine leukaemia virus-related virus in patients with chronic fatigue syndrome in the Netherlands: retrospective analysis of samples from an established cohort. BMJ 2010;340(doi:10.1136/bmj.c1018).
*************************************************
Also for those in the U.K. ♥♥♥

"We are having an independent phlebotomy company draw samples in the UK in the next two weeks if you or others would like to participate send me your addresses and contact info ASAP. Yes we will resume blood draws next week and all 200 participants will be drawn in the next few weeks and all the studies will be done This project is our highest priority! It is going very well and being done efficiently. I hope this addresses your concerns

Best wishes, Judy"'

#60~ Thanks Dr Peterson for EVERTHING~ WPI ♥♥♥

Whittemore Peterson Institute announces search for medical director.

Planned transition paves a smooth path for the fall opening of the new Institute
Reno, Nev.
–
As part of a planned transition, the Whittemore  Peterson Institute (WPI) announces the national search for a full time Medical Director as 

Dr. Daniel Peterson retires from this position.
 

Dr. Peterson, a pioneering physician in describing methods of diagnosing, managing and treating myalgic encephalomyelitis (ME/CFS), was one of the
first physicians to identify this disease in the United States.

“Dr. Peterson was central figure in the establishment of the Whittemore Peterson
Institute,”
remarks Annette Whittemore, founder and president of WPI.
“We are deeply grateful to him for his significant advice and support through
the development of the Institute.”

“I am extremely proud to have been a part of the creation of this medical research
institute which promises to bring exciting new answers to patients with
complex neuro-­‐immune diseases,” states Dr. Peterson who will continue his
internal medicine practice in Incline Village, Nev.

The WPI will soon begin a national search for a new full-­‐time medical director
and practicing physician with extensive experience in infectious disease, clinical
trials and medical management.

The Whittemore Peterson Institute will move into a new 100,000-­‐square-­‐foot
state-­‐of-­‐the art research and medical facility, the Center for Molecular Medicine,
at the University of Nevada in the fall of 2010.
Dr. Judy Mikovits will continue to lead the comprehensive research program at the
WPI, which will be the first in the world dedicated to neuro-­‐immune diseases
integrating patient treatment, basic research, clinical trials and medical education.

To learn more about the institute and ongoing research, please visit
http://www.wpinstitute.org.


For the FULL article download the pdf.

http://www.wpinstitute.org/news/docs/NewMedicalDirector_042810.pdf.

#48~ Maybe I'll do a 3 day Medical Study & Publish it?

If psychiatrists can research retrovirology,
and publish medical research papers on 
the same Hey..maybe I can also?  

Heck, why don't we get
a few photographers in, a philosopher 
or two and make it a real party. Not to 
mention an astronomer, a comedian and 
an arborist to keep it balanced! 
Anyone else in?  Look how much 
"media" attention it would get! 

Aren't you TIRED of listening to False

BAD news every day? Let's go 

"on Strike" and only watch/read

THE GOOD NEWS & The TRUTH

from Now ON.. and "Most Importantly" 
Expose the Imposters & Scammers, OK?

Earlier this month that's exactly what
happened in the U.K. regarding a VERY
Important Medical Issue of 
World Importance.

As opposed to waiting for the REAL
Medical Researchers to complete a
REAL study that would replicate the
findings of a new Retrovirus that has
World Health implications...
...tiny minded folks in the pysch
community tried to whip out a
3 day study & write a paper...

When knowing that to do "REAL Research"
takes TIME and meticulous repetition of
not only the EXACT technique originally
used, but also looking for the same strain
of the retrovirus, and using a pool of
test subjects that have been
drawn from the SAME diagnostic criteria
as the original study was...
or else.. 

this Imposter is just THAT..
..and TOTAL SHAM and a disgrace out 

there to the REAL Psychiatrists that are
trying to do some REAL help for their
patients.. 

Especially, if the psychiatrist
that in this study has been previously
involved in muddy situations of possible
child/patient abuse in the past.

And SHAME on the Newspapers &
Magazines and ezines that have so quickly
"jumped onto" this story instead of doing
PROPER "Investigative Reporting."
Obviously, these issues are what we
call in the industry "rags" and not really
producers of Real NEWS, but rather
gossip and BS attention headline grabbing
garbage.... which result in articles that
match the quality of the 3 day study paper
they were "supposedly" reporting on..
aka "Garbage IN = Garbage OUT."

It's HARD enough to get the money
to get REAL Scientific Research funded..
We do NOT need to waste time nor money
for these sham studies or give them ANY
Credibility.... When you read an article,
read the "fine print" also... the details of
who was involved in the study, did they
use proper techniques and reputable
research partners and take the proper
amount of time and use the proper
controls of whom was involved in the
study ??

Just like everything else in life
"Caveat Emptor" = Buyer Beware.
BE an Informed Reader/Patient
/Traveler/ whatever the topic du jour.

My friend that was a reporter taught 
me WELL..
Just because something is in Print
does NOT mean that is it TRUE...
Just because you hear on the radio
doesn't mean it's TRUE...
Just because you read it on the
internet doesn't mean it's TRUE..
Seeing a pattern here?
You mean to tell me you believe every
stupid commercial and ad you read ??
I don't think so....
Unless you are a scammers Dream?

with THAT.. I will give you the Statement
provided from the Original researchers
that did the Original Study that also
collaborated with the Cleveland Clinic 
and the USA National Cancer Institute.

*******************************
Official Statement from the 
Whittemore Peterson Institute 
Regarding UK Study
 
Wednesday, January 6, 2010 at 1:00pm
FOR IMMEDIATE RELEASE

Frankie Vigil
R&R Partners for
Whittemore Peterson Institute
775-336-4555
frankie.vigil@rrpartners.com

Official Statement from the 

Whittemore Peterson Institute 

Regarding UK Study

The Whittemore Peterson Institute (WPI) 

has reviewed the paper entitled 

“Failure to Detect the Novel Retrovirus 

XMRV in Chronic Fatigue
Syndrome.” This study did not duplicate 

the rigorous scientific techniques used 

WPI, the National Cancer Institute and 

the Cleveland Clinic, therefore it cannot 

be considered a replication study nor can

the results claim to be anything other 

than a failure not just to detect XMRV, 

but also a failure to suggest 

meaningful results.

The scientific methods used by WPI are
very exact and require specific techniques
 to ensure accuracy. Differences in
techniques employed by Erlwein et al.
not only explain their failure to

replicate the WPI study, but also render 

the conclusions meaningless.
These differences include, but are
not limited to the following:

1) blood sample volumes and processing;
2) patient criteria/population differences;
3) number and type of tests done to assure
       accurate results, including white blood 
       cell culture;
4) use of a molecular plasmid control in water
       versus a positive blood sample; and
5) different primer sequences and 
       amplification protocol used to find 
       the virus, which were not 
       validated by a clinical control.


The WPI study was published after 

six months of rigorous review and three 

independent lab confirmations, proving 
that contamination had not taken place 
and that infectious XMRV was present in
67 percent of CFS patients diagnosed 
according to the Canadian and
Fukuda criteria. 

In contrast, this latest study
was published online after only three days
of review. Significant and critical questions
remain as to the status of patient samples
used in the UK study as those samples may
have been confused with fatigued psychiatric
patients, since the UK has relegated
“CFS” patients to psychiatric care and not
traditional medical practices.

“Little is known about the prevalence of
XMRV world-wide, much less the incidence
of XMRV in ME/CFS or prostate cancer”
emphasizes Dr. Judy Mikovits.
“WPI and its NCI collaborators are actively
engaged with international research teams
to investigate these important questions.”

WPI does not recommend the use of
anti-retroviral drugs that have yet to be
proven to be effective in treating XMRV
infection. However, several large
pharmaceutical companies have expressed
interest in developing anti-retroviral and
immune modulating drugs that will
effectively treat XMRV associated 
diseases.

WPI looks forward to the results of other
scientific groups around the world, 

serious about replicating its scientific 

results, by using the same techniques as

WPI and its collaborators. The fact that 
XMRV was detected in 67 percent of the 
CFS samples in the U.S. study determined
a significant association between XMRV
and CFS, demanding a much more 

serious inquiry by responsible health 

agencies around the world as to the 

cause of this debilitating disease.

-###-

Whittemore Peterson Institute
The Whittemore Peterson Institute for
Neuro-Immune Disease exists to bring
discovery, knowledge, and effective 
treatments to patients with illnesses that
are caused by acquired dysregulation of 
the immune system and the nervous 
system, often results in lifelong disease
and disability. 

The WPI is the first institute in the world
dedicated to
X associated neuro-immune disease 
(XAND), and other X associated diseases,
integrating patient treatment, basic and
clinical research and medical education.
*******************************

Wouldn't the world be in even a worse 
mess if all the people from different 
walks of life could do a 3 day Medical study
and then publish them as if they were 
Medical FACT, while trying to negate the 
work of the Real Scientific Researchers
that have devoted their lives to doing 
REAL WORK in any certain specific field
of Medical Research ?

Besides the Investor Scammers let's

start throwing a wider net and keep 

and eye Open for the Scammers in

other areas of life that are also

Important to us also....like our Health !!!

Doesn't matter WHAT country or 

continent they are from... wrong is 

wrong.

As one my old Favorite movies use to say:

"I'm MAD as Hell and I'm 

NOT going to take it any more."

#9- Dr. Judy Mikovits, PhD, to report Oct 19th

Dr. Judy Mikovits, PhD, will present her
important new chronic fatigue syndrome
research findings in Lisbon, Portugal on
Monday, Oct 19, at the Tri-Society Annual
Conference 2009. Dr. Mikovits is
Director of Research at the
Whittemore-Peterson Institute for
Neuro-Immune Disease in Reno, Nevada.

We have been awaiting some research
that will come from them, so this
hopefully will be of some IMPORTANCE
to us.. time will tell... to be continued.

I have just added a video of a presentation
made in 2008 by DR. Daniel Peterson,
on the right under videos, he is the
Peterson of this Institute.
 
WPI.org