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VDB admits doping...?

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Road Cycling
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3 October 2003
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Hb
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  1. "Kyle Legate" <[email hidden]> wrote in message
    "]news:[email hidden]...

    Quoted message said:
    Quoted message said:


    The key, they say, is genetic manipulation. By the Athens Olympics in 2004, dozens if not
    hundreds of athletes are expected to have experimented with the rapidly emerging range of
    gene-altering drugs. Unfettered by fears of being caught they will, according to the experts,
    shatter the accepted limits of human performance.


    No.

    Maybe by the _next_ Athens Olympics, but not these ones. The technology is not there yet. What you
    quoted was an ill-informed piece of fluff.

    Dumbass -

    perhaps, but its being speculated about all over the 'net. If people are thinking about it, that
    means that somewhere, someone is going to try it, sooner rather than later.

    maxmag.maxsportsinternational.com28sci1.htm

    With relatively few old-timers showing an inclination to pump iron three times a week for the rest
    of their of lives, the potential market for an alternative muscle-building drug is clearly enormous.
    Science finally appears close to creating one.

    In separate experiments over the past couple of years at the University of Pennsylvania Medical
    Center in Philadelphia, and University College Medical School in London, as well as the Copenhagen
    Muscle Research Center in Sweden, researchers have tested muscle-building vaccines based on
    engineered genes. Injected into mice, these vaccines have boosted muscle mass in the animals legs by
    15 to 27%. Amazingly, these increases were measurable in only a month or so and did not require any
    exercise at all! Many muscle researchers believe that the first human trials will occur within the
    next couple of years. This could also be a major breakthrough for the treatment of a host of
    degenerative muscle diseases, including the various forms of muscular dystrophy.

    On the down side, it takes little imagination to see the possibilities for abuse of the vaccines by
    healthy young athletes in power sports such as football, weight lifting, sprinting and
    short-distance swimming. Compared with anabolic steroids, a vaccine based on an engineered gene
    would offer some major advantages. It would need to be administered only one time, rather than
    periodically, and it would be essentially undetectable in the body.

    MUSCLE PHYSIOLOGY 101

    A single muscle cell consists of a membrane, many scattered nuclei that contains genes and thousands
    of inner strands called myofibrils. Filling the inside of muscle fiber, the myofibrils can be as
    long as the fiber and are the part that enables the cells to contract forcefully in response to
    nerve impulses. The actual contraction is accomplished by the myofibril tiny component units which
    are called sarcomeres. Within each sarcomere are two proteins, called myosin and actin, whose
    interaction causes contraction of the muscle. Basically, during contraction a sarcomere is shortened
    like a collapsing telescope, as the actin filaments at each end of a central myosin filament slide
    toward to the myosin's center.

    Muscle cells, also known as fibers cannot split themselves to form completely new fibers. A muscle
    can become more massive only when its individual fibers become thicker. What causes this thickening
    is the creation of new myofibrils. The mechanical stresses that exercise exerts on tendons and other
    structures connected to the muscle trigger different biochemical pathways that ultimately cause the
    muscle cells to make more proteins.

    Enormous amounts of these proteins, chiefly myosin and actin, are needed as the cell produces
    additional myofibrils. As muscle cells cannot divide, the new nuclei are donated by so-called
    satellite cells, which are scattered among the many nuclei on the surface of a skeletal muscle
    fiber. Satellite cells proliferate in response to the stresses and wear and tear of exercise. As
    they multiply, some remain as satellites on the fiber, but others become incorporated into it. With
    these additional nuclei, the fiber is able to turn out more proteins and create more myofibrils.

    Rigorous exercise inflicts tiny "micro tears" in muscle fibers. The damaged area attracts the
    satellite cells, which incorporate themselves into the muscle tissue and begin producing proteins to
    fill the gap. Gradually, as more micro tears are repaired in this manner, the overall number of
    nuclei grows, as does the fiber itself (i.e. muscle enlarges).

    One component of the myosin molecule, the so-called heavy chain, determines the functional
    characteristics of the muscle fiber. In an adult, this heavy chain exists in three different forms,
    known as isoforms. These isoforms are designated Type I, Type IIa, and Type IIx, as are the fibers
    that contain them.

    Type I fibers are also known as slow fibers; Type IIa and IIx are referred to as fast fibers. The
    fibers are called slow and fast for good reason; the maximum contraction velocity of a single Type I
    fiber is approximately 1/10th of every Type IIx fiber. The velocity of Type IIa fibers are somewhere
    between those of Type I and IIx. Slow fibers depend more on relatively efficient aerobic exercise
    where as the fast fibers depend more on anaerobic exercise. Thus, slow fibers are important for
    endurance activities and sports such as long distance running, cycling, or swimming, where as fast
    fibers are key to power pursuits such as weight lifting and sprinting.

    The "average" healthy adult has relatively equal numbers of slow and fast fibers in say the
    quadriceps muscle of the thigh. But as a species, humans show a great variation in this regard. A
    person with a predominance of slow fibers would probably become an accomplished marathoner but would
    never get anywhere as a sprinter or power lifter; the opposite would be true of a person with the
    predominance of fast fibers.

    MUSCLE CONVERSION

    When healthy muscles are loaded heavily and repeatedly, as in weight training programs, the number
    of fast IIx fibers declines as they convert to fast IIa fibers. In those fibers, the nuclei stop
    expressing the IIx gene and begin expressing the IIa. If vigorous exercise continues for about a
    month or more, the IIx fibers will completely transform to IIa fibers. At the same time, the fibers
    increase their production of proteins, becoming thicker (hypertrophy).

    CONVERTING SLOW TO FAST?

    Is it possible to convert the slower Type I fibers to faster Type II fibers? In the early 1990's
    there was an indication that a rigorous exercise regimen could convert slow fibers to fast IIa
    fibers. Researchers at the University of Copenhagen Muscle Research Center suggested that a program
    of vigorous weight training supplemented with other forms of anaerobic exercise converts not only
    Type IIx fibers to IIa, but also Type I fibers to IIa. If a certain type of exertion can convert
    some Type I fibers to IIa, we might naturally wonder if some other kind can convert IIa to I. It may
    be possible, but so far no link in human training studies has unambiguously demonstrated such a
    shift. It is true, star endurance athletes such as long-distance runners and swimmers, cyclists and
    cross-country skiers generally have remarkably high proportions (up to 95%, as mentioned earlier) of
    slow Type I fibers in their major muscle groups, such as in the legs. Yet at present we do not know
    whether these athletes were born with such a high percentage Type I fibers and gravitated toward
    sports that take advantage of unusual inborn traits or whether they very gradually increased the
    proportion of Type I fibers in their muscles as they trained over a period of many months or years.

    Researchers have found that hypertrophy from resistance training enlarges Type II fibers twice as
    much as it does type I fibers. Thus, weight training can increase the cross-sectional area of the
    muscle covered by fast fibers without changing the relative ratio between the number of slow and
    faster fibers in the muscle. It is the relative cross-sectional area of the fast and slow fiber that
    determines the functional characteristics of the entire muscle. The more area covered by fast
    fibers, the fast and more powerful the overall muscle will be. So a sprinter at least has the option
    of altering the characteristics of his or her leg muscles by exercising them with weights to
    increase the relative cross section of fast fibers.

    THE ERA OF GENETIC MANIPULATION

    Although certain types of fiber conversion, such as IIa to I appear to be difficult to bring about
    through exercise, the time is rapidly approaching when researchers do have the capability to
    accomplish such conversions easily through genetic techniques. Such genetic manipulations, most
    likely in the form of vaccines that insert artificial genes into the nuclei of muscle cells, will
    almost certainly be the performance enhancing drugs of the future. The tiny snippets of genetic
    material and the proteins that gene therapy will leave behind in the athletes muscle cells may be
    difficult, if not will be impossible, to identify as foreign.

    Gene therapy is now being researched intensively in most developed countries for a host of very good
    reasons. Instead of treating the deficiencies by injecting drugs, doctors will be able to prescribe
    genetic treatments that will induce the bodies own protein-making machinery to produce the proteins
    needed to combat illness. Like ordinary genes, the artificial gene consists of DNA. It can be
    delivered to the body in several ways. Suppose the gene is encoded for one of the many signaling
    proteins or hormones (testosterone or growth hormone) that stimulate muscle growth. The approach
    would be to inject the DNA via vaccine into the muscle. The muscle fibers would then take up the DNA
    and add it to the normal pool of genes.

    This method is not very efficient yet, so researchers often use viruses to carry the gene payload
    into a cells nuclei. A virus is essentially a collection of genes packed in a protein capsule that
    is able to bind to a cell and inject the genes. Scientists replace the viruses own genes with the
    artificial gene (i.e. the muscle growth stimulator gene), which the virus will then efficiently
    deliver to the cells in the body.

    GET PUMPED THE EASY WAY

    It is easy to see how the narcissist would find the drug irresistible. A vaccine to build muscle
    mainly where it was injected, making it possible for even the lazy and uncoordinated to sculpt their
    bodies by doing nothing more strenuous than lifting a hypodermic needle. Big biceps, nice calves and
    big bulging pecs would all be just a few injections away. Of course, an instant physique of this
    kind would not come without a physiological price. To improve performance or look really buff,
    athletes and body builders would probably need to take considerably larger doses than what doctors
    will prescribe for therapy. Thus, they would probably suffer some of the already known or suspected
    side effects for abuse of IGF-1, such as an enlarged heart and possibly cardiac arrest.

    THE GENETICALLY ENGINEERED SUPER ATHLETE

    These techniques will be abused by athletes in the future. Sports officials will be hard-pressed to
    detect the abuse, because the artificial genes will produce proteins that in many cases are
    identical to the normal proteins. Furthermore, only one injection will be needed, minimizing the
    risk of disclosure. It is true that officials would be able to detect the DNA of the artificial gene
    itself, but to do so they would have to know the sequence of the artificial gene, and the esters
    would have to obtain a sample of the tissue containing the DNA. Today, however, biopsies are not
    permitted as part of a routine anti-doping test. For all intents and purposes, gene doping will be
    undetectable.

    <snip><end

  2. Quoted post said:

    Originally posted by Kyle Legate
    Maybe by the _next_ Athens Olympics, but not these ones. The technology is not there yet. What you
    quoted was an ill-informed piece of fluff.

    I have a feeling Kurgmeister is right.

    Tetracycline induceable systems for regulated expression are showing promise. Adeno associated vectors can be made to integrate into the genome, or to just transiently infect:

    DNA Cell Biol. 2002 Dec;21(12):895-913. Related Articles, Links

    Adenovirus and adeno-associated virus vectors.

    Lai CM, Lai YK, Rakoczy PE.

    Centre for Ophthalmology and Visual Science, University of Western Australia, Nedlands, Western Australia.

    Recombinant adenovirus (rAd) and recombinant adeno-associated virus (rAAV) are among the most extensively used vectors in gene therapy studies to date. These two vectors share some similar features such as a broad host range and ability to infect both proliferating and quiescent cells. However, they also possess their own unique set of properties that render them particularly attractive for gene therapy applications. rAd vectors can accommodate larger inserts, mediate transient but high levels of protein expression, and can be easily produced at high titers. Development of gutted rAd vectors has further increased the cloning capacity of these vectors. The gaining popularity of rAAV use in gene therapy can be attributed to its lack of pathogenicity and added safety due to its replication defectiveness, and its ability to mediate long-term expression in a variety of tissues. Site-specific integration, as occurs with wild-type AAV, will be a unique and valuable feature if incorporated into rAAV vectors, further improving their safety. This paper describes these properties of rAd and rAAV vectors, and discusses further development and vector improvements that continue to extend the utility of these vectors, such as cell retargeting by capsid modification, differential transduction by use of serotypes, and extension of the cloning capacity of rAAV vectors by dual vector heterodimerization.

    Hum Gene Ther. 2003 Sep 1;14(13):1265-77.

    Tight positive regulation of transgene expression by a single adenovirus vector containing the rtTA and tTS expression cassettes in separate genome regions.

    Mizuguchi H, Xu ZL, Sakurai F, Mayumi T, Hayakawa T.

    Division of Cellular and Gene Therapy Products, National Institute of Health Sciences, Tokyo 158-8501, Japan. [email hidden]

    We previously developed single adenovirus (Ad) vectors that contained the components for a tetracycline-regulatable gene-expression system in the E1 and E3 deletion regions, and showed that the Ad vectors containing the tet-on system exhibit a much inferior regulation of transgene expression than those containing the tet-off system. In many cases, the tet-on system may be preferable because of its positive regulation of transgene expression. To this end, in the present study, by introducing the latest generation reverse tetracycline-responsive transcriptional activator (rtTA2s-M2 or rtTA2s-S2) and the tetracycline-controlled transcriptional silencer (tTS) into the original tet-on system, we constructed various modified Ad-mediated tet-on systems. Among them, the novel single Ad vector, which contained three heterologous gene-expression cassettes of the gene of interest, rtTA2s-S2, and tTS in the E1 deletion region, the E3 deletion region, and the region between E4 and 3'ITR, respectively, displayed vastly improved doxycycline-inducible gene expression in terms of low basal expression, high induced expression, and high responsiveness to doxycycline both in vitro and in vivo. These results also suggest that the low responsiveness to doxycycline may explain why the original tet-on system in the context of the Ad vector is not effective in vivo. This is the first report describing the cloning of three heterologous gene-expression cassettes into three separate regions of the E1/E3-deleted Ad genome. This improved Ad-mediated tet-on system might be useful for gene therapy and greatly facilitate the analyses of gene function.

    PMID: 12952598 [PubMed - in process]

  3. Kurgan Gringioni said:

    "Kyle Legate" <[email hidden]> wrote in message
    "]news:[email hidden]...

    Quoted message said:
    Quoted message said:


    Yack yack yack, you've already posted this in it's entirety, you don't have to do it again.
    What I would appreciate more is a reference to the peer reviewed literature from the groups
    mentioned above.

    After the scientists injected the virus into the animals' muscles, it infiltrated their cells,
    inserting the epo gene and spurring the cells to pump out the protein. This boosted mouse
    hematocrits (the proportion of the blood volume made up of red blood cells) from 49 per cent to 81
    per cent, while the monkeys' hematocrits rose from 40 per cent to 70 per cent or more (Human Gene
    Therapy, vol 8, p 1797). A single injection elevated hematocrits for over a year in the mice and
    for 12 weeks in the monkeys.

    Researchers at the biotech company Chiron in Emeryville, California, reported similar results in a
    1998 trial that used AAVs to deliver the epo gene to two baboons (Gene Therapy, vol 5, p 665).
    After 10 weeks, their hematocrits had risen from 38 per cent and 40 per cent to 62 and 75 per
    cent, respectively, and stayed at those levels for the entire 28 weeks of the study.


    Read this again and tell me if it's a feasible approach in humans. My original questions all still
    stand, and you have made no effort to answer them.

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