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Coronado Biosciences, Inc. Message Board

  • galaberge galaberge Apr 5, 2013 8:48 AM Flag

    Facts about the tests performed...with Gatorade.


    By Robert W. Summers, M.D., David E. Elliott, M.D., Ph.D., Khurram Qadir, M.D., Joseph F. Urban, Jr., Ph.D.,
    Robin Thompson, M.H.A., and Joel V. Weinstock, M.D.
    Department of Internal Medicine, James A. Clifton Center for Digestive Diseases, University of Iowa Health
    Care, Iowa City, Iowa; and The Immunology & Disease Resistance Laboratory, United States Department of
    Agriculture, Beltsville, Maryland

    Treatment With T. suis

    Adult worms were harvested from the intestinal mucosa of
    colonized pathogen-free pigs, washed in sterile buffers, incubated
    in antibiotics, and then cultured in sterile tissue
    culture medium in the presence of antibiotics (12). The eggs
    deposited by cultured worms were isolated, washed, and
    placed in 0.2% potassium dichromate solution at 22°C for
    5–6 wk with aeration to obtain infective first-stage larvae
    (Fig. 1). Eggs were washed twice in sterile water and resuspended
    in the desired amount of saline based on a calculated
    dose of 2500. Before the study started, the infectivity
    of the eggs was checked by recovery of adult worms in
    pigs inoculated with known numbers of eggs. Approximately
    10–20% of the estimated infective eggs developed
    into adult worms.
    Each egg preparation was rigorously tested to confirm
    absence of bacterial and viral pathogens such as Shigella,
    Salmonella, Campylobacter, Yersinia, enterotoxic Escherichia
    coli, cytomegalovirus, herpes simplex virus, adenovirus,
    and enteroviruses using standard microbiological
    The absence of swine hepatitis E viral genome in the T.
    suis eggs was confirmed by a negative reverse transcription
    polymerase chain reaction (RT-PCR). Total RNA was extracted
    from 100 ml of medium containing T. suis eggs
    using RNA STAT-50LS reagent (Tel-Test, Friendswood,
    TX) as described by the manufacturer (13). Reverse transcription
    was performed for 1 h at 37°C using 200 units of
    MMLV reverse transcriptase in the presence of random
    hexamers as primers (13). This PCR generates

    Sentiment: Strong Buy

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      199 bp detected by 2% agarose gel electrophoresis. Positive control was a swine hepatitis E virus isolate with 98%
      homology with US-1 human hepatitis E virus in the nested
      PCR target region (Y.V. Karetnyi and S.J. Naides, GenBank
      accession number AF097722).
      The eggs were stored in aliquots of 1 ml of water containing
      2500 ova at 4°C until they were administered to the study subjects. The eggs remained viable and were able to
      colonize pigs even after 1 yr of storage.
      Study Protocol
      The University of Iowa Institutional Review Board approved
      the study protocol. In total, we studied seven patients
      with refractory IBD (four with CD and three with UC).
      After baseline studies were performed, selection criteria
      were met (as outlined below), and informed consent was
      obtained, we gave 2500 live T. suis eggs orally with 30 ml
      of Gatorade (Gatorade, Chicago, IL). Patients were followed
      every 2 wk without additional helminth ova therapy for _12
      wk. Baseline medications were continued at the same dose
      throughout the period. Safety was monitored by following
      the patients’ clinical status and laboratory studies at regular intervals (see Assessment of Safety). The baseline and follow-
      up efficacy responses were monitored every 2 wk using
      standard clinical indices (see Measurement of Disease Activity).
      This treatment and observation period was continued
      for at least 12 wk. The numerical results at baseline and at 2, 4, 6, 8, and 12 wk were compared in tables and graphs.
      After the first five patients were treated, no adverse events
      were encountered and all patients seemed to improve. Because
      the response was temporary in some patients, we
      requested and received permission to give repetitive doses.
      We wanted to explore whether multiple doses were safe and
      whether the benefit could be extended. After a treatment and
      observation period of at least 12 wk, two patients with CD
      and two with UC were given additional doses of 2500 ova at 3-wk intervals in a maintenance period. Agai

      Sentiment: Strong Buy

1.90-0.20(-9.52%)Oct 1 4:00 PMEDT

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