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Basis for TSE Blood Tests

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Well-known member
Feb 11, 2005
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Home on the Range, Alberta
I apologize in advance for the length of this posting; however, please take a look at it. Much is said about developing blood tests for TSEs. I have shown below the interconnection of three various researchers and their ideas for blood tests. Dr. Vodyanoy of Auburn Uni. tops them all with his statements found within his patent application.

The use of ultrasound, heat and pressure are all forms of applied energy that excite the metal molecules and cause malformation of healthy prions. However, as Kretschmar states, below, without applying the ultrasound, the healthy proteins, which had misfolded during the procedure, quickly converted back to their healthy shape.

The replication of prion proteins associated with neurodegenerative diseases can take up to 40 years in vivo, but a team led by Dr. Claudio Soto of the Serono Pharmaceutical Research Institute in Geneva has been able to complete the process in a single day. "When an animal or a human being gets infected with the prion, this abnormal protein begins to replicate very slowly and transforms the host's normal protein into abnormal protein," says Soto. "So we thought, why can't we do the same thing in a test tube?"

The abnormal protein grows to form aggregates that remain clumped together for long periods. "The rate of growth depends on the number of units," says Soto. "So we tried letting them grow and then breaking them into little pieces, incubating them again to let them grow, and so on in a cyclical process."

The method is analogous to polymerase chain reaction (PCR), the cyclical technique commonly used to detect DNA by amplification. "You can't use conventional PCR here, because there is no DNA," he added. "We tried to create an amplification system like a PCR, but specially for proteins." Minute quantities of hamster brain tissue infected with scrapie were mixed with normal protein from the brain tissue of healthy hamsters, and ultrasound was used to break apart the prion aggregates (Nature 411:810-3). Detection of the resulting abnormal protein was performed by densitometric analysis. "

Breakage of PrP aggregates is essential for efficient autocatalytic propagation of misfolded prion protein.

Piening N, Weber P, Giese A, Kretzschmar H.

Center for Neuropathology and Prion Research, Ludwig Maximilians University of Munich, Feodor-Lynen-Strasse 23, 81377 Munich, Germany.

The conversion of cellular prion protein (PrP(C)) to the disease-associated misfolded isoform (PrP(Sc)) is an essential process for prion replication. This structural conversion can be modelled in protein misfolding cyclic amplification (PMCA) reactions in which PrP(Sc) is inoculated into healthy hamster brain homogenate, followed by cycles of incubation and sonication. In serial transmission PMCA experiments it has recently been shown that the protease-resistant PrP obtained in vitro (PrPres) is generated by an autocatalytic mechanism. Here, serial transmission PMCA experiments were compared with serial transmission reactions lacking the sonication steps. We achieved approximately 200,000-fold PrPres amplification by PMCA. In contrast, although initial amplification was comparable to PMCA reactions, PrPres levels quickly dropped below detection limit when samples were not subjected to ultrasound. These results indicate that aggregate breakage is essential for efficient autocatalytic amplification of misfolded prion protein and suggest an important role of aggregate breakage in prion propagation.

Dr. Claudio Soto’s process of protein misfolding cyclic amplification , otherwise known as PMCA, used “sonication” to break apart prion aggregates to seed new crystals within the next sample.

“ultrasound was used to break apart the prion aggregates"

Dr. H. Kretzschmar, et al. showed in their research that without the sonication step,

“PrPres levels quickly dropped below detection limit when samples were not subjected to ultrasound”

Dr. Kretschmar also made the statement:

“These results indicate that aggregate breakage is essential for efficient autocatalytic amplification of misfolded prion protein and suggest an important role of aggregate breakage in prion propagation.”

What both of these researchers are speaking of, is the seeding and growing of crystals.

Dr. Vitaly Vodyanoy of Auburn University has also stated within his patent application # 20040137523 : (selected quotes)

“The method can be manipulated by subjecting the sample to heat or pressure, or by carrying out various numbers of seeding steps. Methods for amplification include heat and pressure treatment of a sample. Likewise, subjecting a biological sample to or treating it with metal clusters increases the concentration of proteons.”

“[0044] PNC of roughly 1-2 nm and containing about 40-300 atoms play an important role in capturing hemoglobin released into blood plasma. While released hemoglobin is normally captured by protein haptoglobin and endocytosed by macrophages, released hemoglobin can be collected by PNCs. Roughly 7.times.10.sup.13 PNC are present in each milliliter of human blood, while only 0.003% of the whole pool of PNC is normally linked to proteins and made into proteons. However, a proteon of medium size of 160 nm can collect about 100,000 protein molecules of similar size to hemoglobin. Thus, the strong protein scavenging properties of metal nanoparticles allow them to collect proteins including misfolded hemoglobin (Kristiansen et al. (2001) Nature 409:198).”
(“The PNCs are comprised of metallic nanoclusters.”)…

[0093] Individual metallic nanoparticles had a random crystallographic orientation. Thus, in cases where the particles had become clumped (in some cases, this appeared to involve flocculation, in others some of the particles had sintered together), a polycrystalline aggregate was produced. In contrast, within a number of relatively large (around 10 nm diameter or above) clumps of particles, significant (.about.5-10 nm wide) regions were encountered with a constant crystallographic orientation. However, none of the clumps was a true single crystal. Some of these relatively large clumps contained a number of, as yet unidentified, second phases in addition to .alpha.-Fe and Cu.

[0094] Both Cu and Fe form stable oxides (for example the Gibbs free energy of formation of even the relatively low stability CuO phase is around -127 kJ mol.sup.-1 at 300 K). See Brandes. and Brook (1992) Smithells Metals Reference Book (7.sup.th ed., Butterworth-Heinenmann, Oxford, UK). Furthermore, the initial stages of oxidation of these metals are rapid, even at room temperature. For example, logarithmic oxidation of initially bare iron, at an oxygen partial pressure of only 10 mPa, results in the growth of around 2 nm of oxide, after less than 20 minutes at 300 K. See Kruger, J and Yolken (1964), cited by Lawless. (1974) Rep. Prog. Phys. 37(2):231-316. The presence of non-noble metallic nanoparticles implies that the surrounding organic matrix has either impeded oxygen access to the metallic particles and/or has a significant reducing effect.

[0095] Many of the nanoparticles survived coarsening. The surface energy of the particles provides a driving force for larger particles to cannibalize smaller particles (the surface area to volume ratio for a 1-nm particle is 6.times.10.sup.9 m.sup.-1 and this drops by an order of magnitude for a 10-nm particle). Metallic materials have relatively high solid-vapor interfacial energies (.gamma..sub.SV) and those for copper and .alpha.-iron are around the middle of the range for metallic materials (at .about.2.2 and 3.2 J m.sup.-2, respectively; Murr (1975) Interfacial Phenomena in Metals and Alloys (Addison-Wesley; reprinted by TechBooks, Herdon, Va.)). Thus unless the metal-organic matrix interface has an interfacial energy (.gamma..sub.SM) that is such that .gamma..sub.SM<<.gamma..sub.SV, there would remain a significant thermodynamic driving force for coarsening. Given the kinetics of coarsening, if all that were present were the metallic nanoparticles, room-temperature coarsening would occur at a negligible rate (solid-state sintering involves bulk diffusion, interfacial diffusion, free surface diffusion and evaporation and re-condensation, all of which would be very slow for Cu or .alpha.-Fe at room temperature). See Ashby (1974), Acta Metallurgica 22(3):275-289. See also, Swinkels and Ashby (1981) Acta Metallurgica 29(2):259-281. Although the presence of the organic liquid matrix raises possibilities for mass transport, it appears that the matrix did not provide a path for the rapid transfer of metal atoms since many of the nanoparticles of served in the present work remained extremely fine.”

You can judge for yourself what these three researchers have shown.

When I look at their statements, I see a direct correlation to the pain-staking epidemiological work done by Mark Purdey. The sonic boom of aircraft, which many people scoff at, provided a form of sonication which would break apart protein aggregates allowing for the seeding of more crystals within the brain.

A brain that is super-saturated with rogue metal ions would be a perfect growing medium for prions. Subsequent exposure of these compromised people/cattle/deer/cats etc. to organophosphates which tie up copper, or, exposure to molybdenum which also ties up copper, would cause these mammals to develop TSEs once seeding of crystals was initiated.

Continue to subject them to forms of energy like sonic shock waves, ultrasound, sonication eg: thunder, heat and pressure changes, and the time frame for disease progression would greatly increase.

This is my opinion on this subject, which I am backing up with the above statements. And, which I know because of the knowledge I have from spending the last 2 years studying TSEs and Mark Purdey’s works.

Science has tried to advance on these issues, but I believe most researchers are scared to death of running out from under the umbrella of Prusiner’s infectious hypothesis. If this disease were infectious – it would not require additional energy, ie: sonication, to spread it. This disease is a contamination of the body with highly charged metal ions.

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