01 Oct 2017
File List, RICO
1988 Steere says Lyme is like a B cell leukemia
Fungal Exosomes Inhibit Apoptosis
"Vaccines serve the mfgs, not their victims"
CDC Admits Fraud, 2016
Bush's warcrimes, Oct 2000
Also, IDSA REVIEWS, 1989, See the Duray Article,
because he says the same thing there, too. (Continue,
1992: NASA/NIH/NIC/US Army Top
Pathologist: "In Chronic Lyme victims' cerebrospinal fluid, I see what look like Epstein-Barr
Paul Duray, NASA rotating bioreactors (culturing in zero gravity)
FY 98 INTER-AGENCY AGREEMENT
NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT
NATIONAL INSTITUTES OF HEALTH
OFFICE OF LIFE AND MICROGRAVITY SCIENCES AND APPLICATIONS
NATIONAL AERONAUTICS AND SPACE ADMINISTRATION
8. Assessment of the RWV as a "universal" pathogen culture
, 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
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 1970ís 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.
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
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
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.
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.
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.