05/21/2013 09:15:51
Cryme Trainer (moved)
Non-HLA-linked Diseases
Hurricane Sandy and Mold-Related
illnesses (like LYMErix and Lyme Disease, and CFIDS/FM).
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References for psychotropics-induced brain damage
Older data on the incurability of Relapsing Fever
1986, McSweegan trashes Navy for $$$ for ALDF.com
1988, Dattwyler & about immune-suppressing, seronegative Lyme
1990, CDC: "Diagnose Lyme as if it was Relapsing Fever."
Allen Steere "NeuroLyme won't test positive," 1990.
1992, CDC officer Allen Steere falsifies testing in Europe
1992, CDC patents with SmithKline show 2 kinds of Lyme
Compare the 2 kinds of Lyme in the RICO complaint
1994, CDC's Dearborn Booklet .pdf
CDC's invitation to participate in Dearborn .pdf
Evidence Lyme criminals knew LYMErix produced the same "multisystem disease" as "Chronic Lyme"
LYMErix Damage Coverup
(short)
1998, CIA Oilmen & Israelis plan to overthrow Saddam for the oil.
Bush/Gore Oil/War-(Oct,2000)
Bush's own explainer (Oct
2000) re:
Iraq Oil
IDSA REVIEWS, 1989, See the Duray Article,
because he says the same thing then, too.
1992: NASA/NIH/NIC/US Army Top Pathologist: "In Chronic Lyme victims, I see what look like Epstein-Barr transformed lymphocytes."
FY 98 INTER-AGENCY AGREEMENT
NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT
NATIONAL INSTITUTES OF HEALTH
and
OFFICE OF LIFE AND MICROGRAVITY SCIENCES AND APPLICATIONS
NATIONAL AERONAUTICS AND SPACE ADMINISTRATION
8. Assessment of the RWV as a "universal" pathogen culture system
Primary Investigators: , Laboratory of Cellular and Molecular Biophysics, NICHD, NIH and USAMRIID, Dr. Paul Duray, Department of Pathology, NCI, NIH, and Dr. Michael Bray, United States Army Medical Research Institute of Infectious Diseases (USAMRIID), Ft. Detrick, MD.
Aim of Experiments: To demonstrate the potential ability of the RWV Bioreactor to act as a "universal" pathogen culture system for the primary isolation of previously unrecognized pathogens during outbreaks of emerging disease. By demonstrating the ability of the RWV to culture a variety of known pathogens of different classification, i.e. RNA viruses, retroviruses, DNA viruses, parasites, spirochetes, etc., we hope to demonstrate the applicability of the RWV system for de novo primary pathogen isolation protocols.
BACKGROUND:
At present, the de novo isolation of previously unknown or unrecognized emerging disease pathogens requires elucidating the proper culture media or permissive cell line applicable for growth of the pathogenic agent in culture. A few examples serve to illustrate the difficulty inherent in this process using conventional technology. The 1970’s outbreak of Legionaries disease required months to identify the causative agent as a bacterium. The HIV-1 retrovirus required over 2 years to isolate and in the 1993 outbreak of Myuro Canyon disease in the four-corners region of the United States it took 7 weeks to grow the Hantavirus in culture. Clearly these timelines are unacceptable with respect to many public health threat scenarios for emerging disease agents. By utilizing the ability of the RWV to maintain a normal three-dimensional cytoarchitecture and microenviroment for a number of tissues, the possibility of using human tissue explants for primary pathogen isolation, becomes a distinct possibility.
Experiment Design: Attempts will be made to culture a variety of infectious pathogen in the same simple medium (RPMI-1640, 15% FCS) containing human tonsil tissue explants maintained in the RWV Bioreactor. In addition, a human liver and epithelial tissue equivalent will be formulated from established cells lines grown on Cytodex 3 microcarrier beads. These will be co-cultured with the tonsil explants. Known pathogens which have proved to be difficult to isolate by normal protocols, will be introduced into the RWV culture and allowed to incubate with the tissue and tissue equivalents for two weeks.
Continuing Results:
1. It has been previously demonstrated that the spirochetal organism Borrelia burgdorferi, the agent of Lyme disease, grows and proliferates in this system. The system has been shown to support productive infection with HIV-1 strains. Ebola virus was also shown to the amplified in this culture system.
2. The RWV Bioreactor system has been used to study differences between human Ebola strains and Ebola Reston, a macaque strain. Human exposure to Ebola Reston causes no apparent illness, but has an almost 100% mortality rate in non-human primates (NHP). The system has allowed the demonstration that in human tissues unlike in NHPs, Ebola Reston is unable to infect the intact epithelial ling of human blood vessels and this presumably accounts for the non-pathogenicity of the strain in humans.
3. The RWV Bioreactor has also been shown to support productive infection of Monkeypox orthovirus in human lung tissues. This model will hopefully shed light on the pathogenesis of not only monkeypox but also the related smallpox.
Plans: The system will be used to further explore the tissue tropism of Ebola strains, Reston, Zaire, Sudan, and Gabon. RWV Bioreactor cultures will be used to explore the basic pathogenesis of monkeypox in both human and NHP tissues, with particular reference to postulated cytokine production. If possible, in the scientific and political context, this work will be extended to study smallpox infection in human tissues since this type of experiment has never been done before. The RWV will be used in an attempt to culture Treponema pallidum, the causative agent of syphilis. This organism is notorious for its resistance to culture by conventional methods.
9. Study of Lyme disease in RWV Bioreactor
Primary Investigators: Dr. Paul Duray, Department of Pathology, National Cancer Institute, NIH.
Aim of Experiments: To use the RWV Bioreactor to culture Borrelia burgdorferi, the etiologic agent of Lyme disease, in tissues. Proliferation of the spirochete in tissues is desired so that the infection process and ensuing disease progression can be studied in an in vitro model. The model could be used to study the infection process and understand basic questions about the parasite, such as whether it actually infects cells or whether it replicates in intercellular spaces.
BACKGROUND:
Lyme disease is a multisystem inflammatory disease caused by infection with the tick-borne spirochete Borrelia burgdorferi and is the most common vector-borne infection in the United States. Borrelia burgdorferi, is able to persistently infect humans and animals for months or years in the presence of an active immune response. It is not known how the organisms survive immune attack in the mammalian host. There is not an appropriate in vitro model in which to study the progression of Lyme disease in humans or an animal model that presents all the symptoms of human disease.
Experiment Design: Tissue biopsies from patients with Lyme disease will be cultured in RWV Bioreactors to see if proliferation of spirochetes in primary infected tissue can be seen. The RWV will also be used to culture normal human tissues with Borrelia, the Lyme disease parasite, to study the infection process. Media other than BSK will be used to determine whether Borrelia is replicating in the medium in cultures or in the tissue itself, since it is known that Borrelia does not replicate in media other than BSK.
Preliminary Results:
1. Very good replication of Borrelia has been seen tonsils infected with the spirochetes. Infection has been confirmed by silver staining of tissue sections and by PCR.
2. Cultures of tonsils infected with Borrelia in media other than BSK show good replication, suggesting that the spirochete replicates in the tissue blocks and not in the medium.
Plans: Methods of quantitation of spirochete load in infected tissues are being researched so that better methods of comparison are available. With this model of infection we hope to begin studying the differences between infectious and non-infectious Borrelia. We plan to look for genetic variation of infectious spirochetes during a long-term culture.