J Biol Chem. 2008 Mar 10
Manganese binding to the prion protein.
Brazier MW, Davies P, Player E, Marken F, Viles JH, Brown DR.
Biology and Biochemsitry, University of Bath, Bath BA2 7AY.
There is considerable evidence that the prion protein binds copper. However, there have also been suggestions that PrP binds manganese. We used isothermal titration calorimetry to identify the manganese binding sites in wild-type mouse PrP. The protein showed two manganese binding sites with affinities that would bind manganese at concentrations of 63 M and 200 M at pH5.5. This indicates that PrP binds manganese with similar affinity to other know manganese binding proteins. Further study indicated that the main manganese binding site is associated with H95 in the so called "5th site" normally associated with copper binding. Additionally, it was shown that occupancy by copper does not prevent manganese binding. Under these conditions, manganese binding results in an altered conformation of PrP, displacement of copper and altered redox chemistry of the metal-protein complex. Cyclic voltammetric measurements suggest complex redox chemistry involving Mn bound to PrP while Cu bound PrP is able to undergo fully reversible electron cycling. Additionally, Mn binding to PrP converts it to a form able to catalyse aggregation of metal free PrP. These results further support the notion that manganese binding could cause conformation change in PrP and trigger changes in the protein similar to those associated with prion disease.
PMID: 18332141
Listen Matthew -- your conventional transmission theory is all about implying and could's as well.
20 years of testing?????
J Occup Health. 2008 Jan;50(1):1-6.
Body distribution of inhaled fluorescent magnetic nanoparticles in the mice.
Kwon JT, Hwang SK, Jin H, Kim DS, Minai-Tehrani A, Yoon HJ, Choi M, Yoon TJ, Han DY, Kang YW, Yoon BI, Lee JK, Cho MH.
Laboratory of Toxicology, College of Veterinary Medicine, Seoul National University, Seoul, Korea.
Reducing the particle size of materials is an efficient and reliable tool for improving the bioavailability of a gene or drug delivery system. In fact, nanotechnology helps in overcoming the limitations of size and can change the outlook of the world regarding science. However, a potential harmful effect of nanomaterial on workers manufacturing nanoparticles is expected in the workplace and the lack of information regarding body distribution of inhaled nanoparticles may pose serious problem. In this study, we addressed this question by studying the body distribution of inhaled nanoparticles in mice using approximately 50-nm fluorescent magnetic nanoparticles (FMNPs) as a model of nanoparticles through nose-only exposure chamber system developed by our group. Scanning mobility particle sizer (SMPS) analysis revealed that the mice were exposed to FMNPs with a total particle number of 4.89 x 10(5) +/- 2.37 x 10(4)/cm(3) (low concentration) and 9.34 x 10(5) +/- 5.11 x 10(4)/cm(3) (high concentration) for 4 wk (4 h/d, 5 d/wk). The body distribution of FMNPs was examined by magnetic resonance imaging (MRI) and Confocal Laser Scanning Microscope (CLSM) analysis. FMNPs were distributed in various organs, including the liver, testis, spleen, lung and brain. T2-weighted spin-echo MR images showed that FMNPs could penetrate the blood-brain-barrier (BBB). Application of nanotechnologies should not produce adverse effects on human health and the environment. To predict and prevent the potential toxicity of nanomaterials, therefore, extensive studies should be performed under different routes of exposure with different sizes and shapes of nanomaterials.
PMID: 18285638
Sorry, but the link for this paper wouldn't open up for me today. Do a google search.Toxicol Sci. 2006 Jan;89(1):338-47. Epub 2005 Oct 19. Links
Erratum in:
Toxicol Sci. 2006 Mar;90(1):267.
Toxicity and tissue distribution of magnetic nanoparticles in mice.
Kim JS, Yoon TJ, Yu KN, Kim BG, Park SJ, Kim HW, Lee KH, Park SB, Lee JK, Cho MH.
Laboratory of Toxicology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University, Seoul 151-742, Korea.
The development of technology enables the reduction of material size in science. The use of particle reduction in size from micro to nanoscale not only provides benefits to diverse scientific fields but also poses potential risks to humans and the environment. For the successful application of nanomaterials in bioscience, it is essential to understand the biological fate and potential toxicity of nanoparticles. The aim of this study was to evaluate the biological distribution as well as the potential toxicity of magnetic nanoparticles to enable their diverse applications in life science, such as drug development, protein detection, and gene delivery. We recently synthesized biocompatible silica-overcoated magnetic nanoparticles containing rhodamine B isothiocyanate (RITC) within a silica shell of controllable thickness [MNPs@SiO2(RITC)]. In this study, the MNPs@SiO2(RITC) with 50-nm thickness were used as a model nanomaterial. After intraperitoneal administration of MNPs@SiO2(RITC) for 4 weeks into mice, the nanoparticles were detected in the brain, indicating that such nanosized materials can penetrate blood-brain barrier (BBB) without disturbing its function or producing apparent toxicity. After a 4-week observation, MNPs@SiO2(RITC) was still present in various organs without causing apparent toxicity. Taken together, our results demonstrated that magnetic nanoparticles of 50-nm size did not cause apparent toxicity under the experimental conditions of this study.
PMID: 16237191
J Inorg Biochem. 2001 Dec 1;87(3):125-7.
Prion diseases: copper deficiency states associated with impaired nitrogen monoxide or carbon monoxide transduction and translocation.
Sorenson JR.
Department of Pharmaceutical Sciences, Division of Medicinal Chemistry, Slot 522, University of Arkansas College of Pharmacy, University of Arkansas Medical Sciences Campus, 4301 West Markham Street, Little Rock, AR 72205-7122, USA. [email protected]
Literature concerning prion diseases and Cu metabolism was examined to determine merits of various suggestions concerning the relationship between these diseases and altered Cu metabolism. There are a number of recent suggestions that the normal non-pathogenic form of the prion protein (PrP(C)) contains Cu while the abnormal pathogenic form of this protein, PrP(SC), lacks Cu. Results of experiments showing oxidant sensitivity in the presence of ionically bonded Cu and millimolar concentrations of hydrogen peroxide were found to lack relevance. Demonstrating superoxide disproportionation and a correlation with cellular Cu2Zn2SOD activity is relevant and consistent with a role for PrP(C) in Cu endocytosis. There are also a number of recent suggestions that PrP(C) has a role in nerve transmission. Serum from mice that lack cellular PrP(C) was found to have an elevated Cu content consistent with a response to overcome an inflammatory disease. Attempts to induce a 'transmissible' form of prion disease requiring intracerebral injections of somewhat purified brain homogenates were found lacking in support for an etiology occurring as the result of oral ingestion of supposedly 'infected' tissues. It is suggested that PrP(C) is a normal Cu-dependent cuproglycoprotein of unknown function that may have a role in facilitating normal nitrogen monoxide- or carbon monoxide-mediated biochemistry.
PMID: 11730893
The have, in fact, burned the tissue to temperatures of 600 degress Celcius (thus destroying all PROTEINs) and still been able to "transmit" disease by intracranial injection - demonstrating a reverse concept: "that proteins are NOT the transmissible agent"
The ashed protein study:
Proc Natl Acad Sci U S A. 2000 Mar 28;97(7):3418-21.
New studies on the heat resistance of hamster-adapted scrapie agent: threshold survival after ashing at 600 degrees C suggests an inorganic template of replication.
Brown P, Rau EH, Johnson BK, Bacote AE, Gibbs CJ Jr, Gajdusek DC.
Laboratory of Central Nervous System Studies, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892, USA. [email protected]
One-gram samples from a pool of crude brain tissue from hamsters infected with the 263K strain of hamster-adapted scrapie agent were placed in covered quartz-glass crucibles and exposed for either 5 or 15 min to dry heat at temperatures ranging from 150 degrees C to 1,000 degrees C. Residual infectivity in the treated samples was assayed by the intracerebral inoculation of dilution series into healthy weanling hamsters, which were observed for 10 months; disease transmissions were verified by Western blot testing for proteinase-resistant protein in brains from clinically positive hamsters. Unheated control tissue contained 9.9 log(10)LD(50)/g tissue; after exposure to 150 degrees C, titers equaled or exceeded 6 log(10)LD(50)/g, and after exposure to 300 degrees C, titers equaled or exceeded 4 log(10)LD(50)/g. Exposure to 600 degrees C completely ashed the brain samples, which, when reconstituted with saline to their original weights, transmitted disease to 5 of 35 inoculated hamsters. No transmissions occurred after exposure to 1, 000 degrees C. These results suggest that an inorganic molecular template with a decomposition point near 600 degrees C is capable of nucleating the biological replication of the scrapie agent.
PMID: 10716712
""Even enzyme-mimicking inorganic materials can be made by template catalysts such as amorphous metal oxides and zeolites
http://bubbaepx.stmhost.com/2008/03/09/oral-cancer-in-us-soldiers-soldiers-vs-oral-sex-and-gardasil/
March 9, 2008
http://ahrp.blogspot.com/2007/08/oral-cancer-in-us-soldiers-soldiers-vs.html
Oral Cancer in US Soldiers Soldiers vs Oral Sex and Gardasil
ALLIANCE FOR HUMAN RESEARCH PROTECTION, August 27, 2007
A report in the Arizona Daily Star describes in gruesome details of "a bizarrely aggressive oral cancer rarely seen by doctors."
Soldiers who had served in combat zones in Iraq are dying of cancer of the mouth, which until now, overwhelmingly strikes smokers, drinkers and tobacco chewers. But the soldiers who are afflicted with cancers of the mouth do not fit that at risk profile: "These are kids 19, 20 and 21 getting all kinds of cancers. The Walter Reed Army Medical Center cancer ward is packed full with them." The doctors are stumped: "Jim's doctors didn't know why he would get this kind of cancer — they had no answers for us."
The prime causal suspect in the minds of many victims — and some scientists–for the alarming number of Iraq war vets who are dying at record speed " is what's known as depleted uranium (UD)— the radioactive chemical prized by the military for its ability to penetrate armored vehicles. When munitions explode, the substance hits the air as fine dust, easily inhaled.
Last month, the Iraqi environment minister blamed the tons of the chemical dropped during the war's "shock and awe" campaign for a surge of cancer cases across the country.
The Daily Star reports that the ranks of sickened and dying Iraq war vets and their families who believe exposures to toxic poisons in the war zone are behind their illnesses — –is growing. But the military brass withholds information and denies the link repeating the Vietnam Agent Orange scenario: "the number of these cancers remains undisclosed, with military officials citing patient privacy issues, as well as lack of evidence the cases are linked to conditions in the war zone. The U.S. Congress has ordered a probe of suspect toxins and may soon begin widespread testing of our armed forces."
Bloomberg News reports that Merck, a major contributor to the University of Texas M.D. Anderson Cancer Center, and two of its researchers are promoting stepped up studies of the HPV vaccine in boys "to expand the vaccine's use."
The two researchers, Drs. Erich Sturgis and Paul Cinciripini provide the marketing pitch for vaccinating boys with the HPV vaccine in an upcoming article in the journal, Cancer. [1] They claim that oral sex "Changing sexual practices such as more frequent oral sex in adolescents and young adults could contribute to an increase in oncogenic HPV-associated oropharyngeal cancers," researchers said in the report.
Notice, "could contribute" rather than has been proven to contribute….
Indeed, although the HPV vaccine is being offered to males in Australia, Mexico, there is, as yet, no clinical proof that it works to prevent HPV infection in men: " we have no data to confirm that, and we won't have any in the near future," says Debbie Saslow, PhD, of the American Cancer Society.
http://www.webmd.com/sexual-conditions/hpv-genital-warts/news/20070827/hpv-linked-to-throat-cancer-trend?src=RSS_PUBLIC
Clearly, lack of evidence does nothing to prevent researchers, with financial ties to vaccine manufacturers, from making
unsubstantiated claims:
"The best way to reduce cancer-causing HPV is to widen the pool of children vaccinated with Gardasil."
This is a blatant example of academics promoting corporate marketing claims to dictate medical practice. Industry analysts provide a more convincing motivation for the promoting expanded use of the vaccine. By adding boys to the vaccine pool, Gardasil may generate more than $3 billion in annual sales. Not surprising, Merck's spokeswoman said Merck is studying the shot in boys and plans to seek U.S. approval for that use.
Reference:
[1] Sturgis EM, Cinciripini PM "Trends in Head and Neck Cancer Incidence in Relation to Smoking Prevalence: An Emerging
Epidemic of Human Papillomavirus-Associated Cancers?" Cancer 2007;110.
United States Patent Application 20080108085
Kind Code A1
Enari; Masato ; et al. May 8, 2008
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Method and Detection of the Presence of Prions Protein
Abstract
The invention relates to methods for determining the presence of prions in a tissue/organ or fluid therefrom; said method comprising the steps of: contacting the tissue/organ with one or more devices, wherein said devices are capable of binding prions; removing said devices from contact with said tissue/organ; determining if said devices are binding prions wherein the device is contacted with the tissue/organ for 120 minutes.
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Inventors: Enari; Masato; (Tokyo, JP) ; Flechsig; Eckhard; (Wurzburg, DE) ; Collinge; John; (London, GB) ; Weissmann; Charles; (London, GB)
Claims:
1. A method for detecting the presence of prions in a tissue/organ; said method comprising the steps of: (a) contacting the tissue/organ with a device, wherein said device is capable of binding prions; and (b) removing said device from contact with said tissue/organ; and (c) determining if said device is binding prions.
2. A non-invasive method for detecting the presence of prions in a tissue/organ; said method comprising the steps of: (a) contacting the tissue/organ with a device, wherein said device is capable of binding prions; (b) removing said device from contact with said tissue/organ; and (c) determining if said device is binding prions.
3. A method according to claim 1 wherein the device is capable of preserving prions against degradation.
4. A method according to claim 1 wherein the tissue/organ is a mammalian tissue/organ.
5. A method according to claim 4 wherein the tissue/organ is a livestock or a human tissue/organ.
6. A method according to claim 1 wherein the tissue/organ is an tissue/organ in which prions accumulate.
7. A method according to claim 6 wherein the tissue/organ is selected from brain, spleen, lymph node or tonsil.
8. A method according to claim 1 wherein the device comprises metal.
9. A method according to claim 8 wherein the metal comprises one or more metal(s) selected from the group consisting of steel, stainless steel, silver, gold or combinations thereof.
[0004] Usually, diagnosis in humans relies on histopathology and immunohistochemical determination. Further methods for the diagnosis of prion infection require invasive procedures such as brain or tonsil biopsies. Homogenates of these biopsies are injected into the brains of test animals such as mice. If the test animals develop clinical symptoms of prion infection then the brain of the test animal is further examined to confirm that prions are present. Problems associated with this method are that prions contained within the biopsies are subject to degradation. Consequently, infectivity is usually lost within 24 hours.
...
[0006] The present invention provides methods for the detection of prions in a tissue/organ or fluid therefrom. The methods use a device such as a metal wire that is contacted with the tissue/organ. Surprisingly, the device is capable of binding prions within 5 minutes. The device is then removed from contact with the tissue/organ. Surprisingly, the device is able to preserve prions against degradation for greater than 3 days. Using prior art methods, prions degrade after only 24 hours. To determine if the device is binding prions, a number of different methods can be used as discussed below. Since prions bind to the device much faster than previously known, diagnosis of prion infection is significantly quicker than prior art methods.
[0007] According to the first aspect of the present invention, there is provided a method for detecting the presence of prions in a tissue/organ; said method comprising the steps of: contacting the tissue/organ with a device, wherein said device is capable of binding prions; removing said device from contact with said tissue/organ; and determining if said device is binding prions.
[0008] According to a second aspect of the present invention, there is provided a non-invasive method for determining the presence of prions in a tissue/organ; said method comprising the steps of: contacting the tissue/organ with a device, wherein said device is capable of binding prions; removing said device from contact with said tissue/organ; and determining if said device is binding prions. Preferably, said intact tissue/organ is left at least substantially intact by said non-invasive method.
[0009] The device used in the methods of the present invention advantageously preserves prions against degradation.
[0012] The device of the present invention may comprise one or more metals or may comprise plastic such as polystyrene, or glass. It is surprisingly disclosed herein that these materials bind prion protein. Preferably, the device of the present invention may comprise one or more metals. Preferably, the metal is any one or more of the metals selected from the group consisting of steel, stainless steel, silver, gold or combinations thereof. More preferably, the metal is stainless steel.
[0013] Advantageously, the device of the present invention may comprise one or more wires or spheres of diameter less than 5 mm, preferably less than 1 mm, preferably having dimensions as mentioned in the Examples section. Preferably, the device comprises one or more metal wires.
....
[0072] The binding between metals and prions may occur by any form of binding capable of occurring between metals and proteins such as covalent, ionic, Van Der Waals, transient or reversible association, or any other forms of binding interaction.
Preserving Prions
[0073] As used herein, the term "preserving prions" refers to the surprising finding disclosed in the present invention that when prions bind to metal surfaces they are preserved. As used herein, the term "preserved" means that the prions bound to the metal surface are protected against degradation and thus remain infective for a period of time that is longer than would normally be expected. For example, using prior art methods, prions injected into brain remain infective for about 24 hours only. Using the methods of the present invention, prions bound to a metal surface are advantageously preserved in barin for at least 3 days.
[0074] Advantageously, the device may be incubated at a temperature of about -20.degree. C. to further preserve the prions. The preservation may be further enhanced by any action which helps protect prions against degradation such as preventing prions from contacting proteases or preventing prions from contacting phagocytic cells.
[0079] Preferably, the device comprises one or more needles, spatula, pins, wires or spheres. More preferably, the device comprises one or more wires. Most preferably, the device comprises one or wires each measuring about 0.15 mm in diameter and 5 mm in length, such as stainless steel suture monofilament wire available from Braun MelsungerAG, Germany.
.....
[0291] Why are wire-bound prions as infectious as concentrated homogenates? Upon intracerebral inoculation with brain homogenate, infectivity is rapidly distributed throughout the mouse (18) and after 4 days or less prions are no longer detectable in the brain (19). Perhaps wire-bound prions are more stable and can therefore act over a longer period of time.
