19 results on '"Priola, Suzette A."'
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2. Prophylactic and therapeutic effects of phthalocyanine tetrasulfonate in scrapie-infected mice
- Author
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Priola, Suzette A., Raines, Anne, and Caughey, Winslow
- Subjects
Therapeutics -- Physiological aspects ,Therapeutics -- Health aspects ,Phthalocyanins -- Physiological aspects ,Phthalocyanins -- Testing ,Pyrrole -- Physiological aspects ,Pyrrole -- Testing ,Encephalopathy -- Health aspects ,Encephalopathy -- Causes of ,Encephalopathy -- Research ,Encephalopathy -- Care and treatment ,Encephalopathy -- Prevention ,Rodents as laboratory animals -- Research ,Airborne infection -- Research ,Airborne infection -- Causes of ,Health - Published
- 2003
3. The Size and Stability of Infectious Prion Aggregates Fluctuate Dynamically during Cellular Uptake and Disaggregation
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Shoup, Daniel and Priola, Suzette A.
- Abstract
Prion diseases arise when PrPSc, an aggregated, infectious, and insoluble conformer of the normally soluble mammalian prion protein, PrPC, catalyzes the conversion of PrPCinto more PrPSc, which then accumulates in the brain leading to disease. PrPScis the primary, if not sole, component of the infectious prion. Despite the stability and protease insensitivity of PrPScaggregates, they can be degraded after cellular uptake. However, how cells disassemble and degrade PrPScis poorly understood. In this work, we analyzed how the protease sensitivity and size distribution of PrPScaggregates from two different mouse-adapted prion strains, 22L, that can persistently infect cells and 87V, that cannot, changed during cellular uptake. We show that within the first 4 h following uptake large PrPScaggregates from both prion strains become less resistant to digestion by proteinase K (PK) through a mechanism that is dependent upon the acidic environment of endocytic vesicles. We further show that during disassembly, PrPScaggregates from both strains become more resistant to PK digestion through the apparent removal of protease-sensitive PrPSc, with PrPScfrom the 87V strain disassembled more readily than PrPScfrom the 22L strain. Taken together, our data demonstrate that the sizes and stabilities of PrPScfrom different prion strains change during cellular uptake and degradation, thereby potentially impacting the ability of prions to infect cells.
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- 2021
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4. Relative Abundance of apoE and Aβ1–42 Associated with Abnormal Prion Protein Differs between Creutzfeldt-Jakob Disease Subtypes
- Author
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Moore, Roger A., Choi, Young Pyo, Head, Mark W., Ironside, James W., Faris, Robert, Ritchie, Diane L., Zanusso, Gianluigi, and Priola, Suzette A.
- Abstract
Aggregated and protease-resistant mammalian prion protein (PrPSc) is the primary protein component of infectious prions. Enriched PrPScpreparations are often used to study the mechanisms that underly prion disease. However, most enrichment procedures are relatively nonspecific and tend to yield significant amounts of non-PrPSccomponents including various proteins that could confound functional and structural studies. It is thus important to identify these proteins and assess their potential relevance to prion pathogenesis. Following proteinase K treatment and phosphotungstic acid precipitation of brain homogenate, we have used mass spectrometry to analyze the protein content of PrPScisolated from prion-infected mice, multiple cases of sporadic Creutzfeldt-Jakob disease (sCJD), and human growth hormone associated cases of iatrogenic CJD (iCJD). Creatine kinase was the primary protein contaminant in all PrPScsamples, while many of the other proteins identified were also found in non-CJD controls, which suggests that they are not CJD specific. Interestingly, the Alzheimer’s disease associated peptide amyloid β 1–42 (Aβ1–42) was identified in the majority of the sCJD cases as well as non-CJD age-matched controls, while apoliprotein E was found in greater abundance in the sCJD cases. By contrast, while some of the iCJD cases showed evidence of higher molecular weight Aβ oligomers, monomeric Aβ1–42 peptide was not detected by immunoblot, and only one case had significant levels of apolipoprotein E. Our data are consistent with the age-associated deposition of Aβ1–42 in older sporadic CJD and non-CJD patients and suggest that both apolipoprotein E and Aβ1–42 abundance can differ depending upon the type of CJD.
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- 2016
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5. Proteomics Analysis of Amyloid and Nonamyloid Prion Disease Phenotypes Reveals Both Common and Divergent Mechanisms of Neuropathogenesis
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Moore, Roger A., Sturdevant, Dan E., Chesebro, Bruce, and Priola, Suzette A.
- Abstract
Prion diseases are a heterogeneous group of neurodegenerative disorders affecting various mammals including humans. Prion diseases are characterized by a misfolding of the host-encoded prion protein (PrPC) into a pathological isoform termed PrPSc. In wild-type mice, PrPCis attached to the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor and PrPSctypically accumulates in diffuse nonamyloid deposits with gray matter spongiosis. By contrast, when mice lacking the GPI anchor are infected with the same prion inoculum, PrPScaccumulates in dense perivascular amyloid plaques with little or no gray matter spongiosis. In order to evaluate whether different host biochemical pathways were implicated in these two phenotypically distinct prion disease models, we utilized a proteomics approach. In both models, infected mice displayed evidence of a neuroinflammatory response and complement activation. Proteins involved in cell death and calcium homeostasis were also identified in both phenotypes. However, mitochondrial pathways of apoptosis were implicated only in the nonamyloid form, whereas metal binding and synaptic vesicle transport were more disrupted in the amyloid phenotype. Thus, following infection with a single prion strain, PrPCanchoring to the plasma membrane correlated not only with the type of PrPScdeposition but also with unique biochemical pathways associated with pathogenesis.
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- 2014
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6. The role of the prion protein membrane anchor in prion infection
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Priola, Suzette A. and McNally, Kristin L.
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In transmissible spongiform encephalopathies (TSE or prion diseases) such as sheep scrapie, bovine spongiform encephalopathy and human Creutzfeldt-Jakob disease, normally soluble and protease-sensitive prion protein (PrP-sen or PrPC) is converted to an abnormal, insoluble and protease-resistant form termed PrP-res or PrPSc. PrP-res/PrPSc is believed to be the main component of the prion, the infectious agent of the TSE/prion diseases. Its precursor, PrP-sen, is anchored to the cell surface at the C-terminus by a co-translationally added glycophosphatidyl-inositol (GPI) membrane anchor which can be cleaved by the enzyme phosphatidyl-inositol specific phospholipase (PIPLC). The GPI anchor is also present in PrP-res, but is inaccessible to PIPLC digestion suggesting that conformational changes in PrP associated with PrP-res formation have blocked the PIPLC cleavage site. Although the GPI anchor is present in both PrP-sen and PrP-res, its precise role in TSE diseases remains unclear primarily because there are data to suggest that it both is and is not necessary for PrP-res formation and prion infection.
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- 2009
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7. Cyclic Tetrapyrrole Sulfonation, Metals, and Oligomerization in Antiprion Activity
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Caughey, Winslow S., Priola, Suzette A., Kocisko, David A., Raymond, Lynne D., Ward, Anne, and Caughey, Byron
- Abstract
ABSTRACTCyclic tetrapyrroles are among the most potent compounds with activity against transmissible spongiform encephalopathies (TSEs; or prion diseases). Here the effects of differential sulfonation and metal binding to cyclic tetrapyrroles were investigated. Their potencies in inhibiting disease-associated protease-resistant prion protein were compared in several types of TSE-infected cell cultures. In addition, prophylactic antiscrapie activities were determined in scrapie-infected mice. The activity of phthalocyanine was relatively insensitive to the number of peripheral sulfonate groups but varied with the type of metal bound at the center of the molecule. The tendency of the various phthalocyanine sulfonates to oligomerize (i.e., stack) correlated with anti-TSE activity. Notably, aluminum(III) phthalocyanine tetrasulfonate was both the poorest anti-TSE compound and the least prone to oligomerization in aqueous media. Similar comparisons of iron- and manganese-bound porphyrin sulfonates confirmed that stacking ability correlates with anti-TSE activity among cyclic tetrapyrroles.
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- 2007
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8. DNA Aptamers That Bind to PrPCand Not PrpScShow Sequence and Structure Specificity
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Takemura, Kaori, Wang, Ping, Vorberg, Ina, Surewicz, Witold, Priola, Suzette A., Kanthasamy, Anumantha, Pottathil, Ravi, Chen, Shu G., and Sreevatsan, Srinand
- Abstract
DNA aptamers were selected against recombinant human (rhu) cellular prion protein (PrPc) 23–231 by systematic evolution of llgands via a systematic evolution of ligands by exponential (SELEX) enrichment procedure using lateral flow chromatography. The SELEX procedure was performed with an aptamer library consisting of a randomized 40-nucleotide core flanked by 28-mer primer-binding sites that, theoretically, represented approximately 1024distinct nucleic acid species. Sixty nanograms of rhuPrPc23–231 immobilized in the center of a lateral flow device was used as the target molecule for SELEX. At the end of 6 iterations of SELEX, 13 distinct candidate aptamers were Identified, of which, 3 aptamers represented 32%, 8%, and 5% of the sequences respectively. Eight aptamers, including the three most frequently occurring candidates, were selected for further evaluation. Selected aptamers bound to rhuPrPc23–231 at 10–6M to 10–8M concentrations. Two of the eight aptamers bound at higher concentrations to rhuPrPc90–231. Theoretical thermodynamic modeling of selected aptamer sequences Identified several common motifs among the selected aptamers that could play a role in PrP binding. Binding affinity to rhuPrPc23–231 was both aptamer sequence and structure dependent. Further, selected aptamers bound to mammalian PrPs derived from brain of healthy sheep, calf, piglet, and deer, and to PrPcexpressed in mouse neuroblastoma cells. None of the aptamers bound to proteinase K-digested scrapie-infected mouse neuroblastoma cells or untreated PrP-null cells, which further confirmed the PrPcspecificity of the aptamers. In summary, we enriched and selected DNA aptamers that bind specifically to rhuPrPcand mammalian PrPcwith varying affinities and can be applied to biological samples for PrPcenrichment and as diagnostic tools in double ligand assay systems.
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- 2006
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9. Molecular Basis of Scrapie Strain Glycoform Variation*
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Vorberg, Ina and Priola, Suzette A.
- Abstract
Transmissible spongiform encephalopathies (TSE) are characterized by the conversion of a protease-sensitive host glycoprotein, prion protein or PrP-sen, to a protease-resistant form (PrP-res). PrP-res molecules that accumulate in the brain and lymphoreticular system of the host consist of three differentially glycosylated forms. Analysis of the relative amounts of the PrP-res glycoforms has been used to discriminate TSE strains and has become increasingly important in the differential diagnosis of human TSEs. However, the molecular basis of PrP-res glycoform variation between different TSE agents is unknown. Here we report that PrP-res itself can dictate strain-specific PrP-res glycoforms. The final PrP-res glycoform pattern, however, can be influenced by the cell and significantly altered by subtle changes in the glycosylation state of PrP-sen. Thus, strain-specific PrP-res glycosylation profiles are likely the consequence of a complex interaction between PrP-res, PrP-sen, and the cell and may indicate the cellular compartment in which the strain-specific formation of PrP-res occurs.
- Published
- 2002
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10. Antiaggregating Antibody Raised Against Human PrP 106-126 Recognizes Pathological and Normal Isoforms of the Whole Prion Protein
- Author
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Hanan, Eilat, Priola, Suzette A., and Solomon, Beka
- Abstract
Antibodies to the prion protein (PrP) have been critical to the neuropathological and biochemical characterization of PrP-related degenerative diseases in humans and animals. Although PrP is highly conserved evolutionarily, there is some sequence divergence among species; as a consequence, anti-PrP antibodies have a wide spectrum of reactivity when challenged with PrP from diverse species. We have produced an antibody [monoclonal antibody (mAb) 2-40] raised against a synthetic peptide corresponding to residues (106-126 of human PrP and have characterized it by epitope mapping, Western immunoblot analysis, and immunohistochemistry. The antibody recognizes not only human PrP isoforms but also pathological PrP from all species tested (i.e., sheep, hamsters, and mice). Together with the fact that it recognizes the whole PrP in both cellular and scrapie isoforms, mAb 2-40 may be helpful in studying conformational changes of the PrP, as well as establishing a possible connection between human and animal diseases.
- Published
- 2001
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11. Deletion of ß-Strand and a-Helix Secondary Structure in Normal Prion Protein Inhibits Formation of Its Protease-Resistant Isoform
- Author
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Vorberg, Ina, Chan, Kaman, and Priola, Suzette A.
- Abstract
ABSTRACTA fundamental event in the pathogenesis of transmissible spongiform encephalopathies (TSE) is the conversion of a normal, proteinase K-sensitive, host-encoded protein, PrP-sen, into its protease-resistant isoform, PrP-res. During the formation of PrP-res, PrP-sen undergoes conformational changes that involve an increase of ß-sheet secondary structure. While previous studies in which PrP-sen deletion mutants were expressed in transgenic mice or scrapie-infected cell cultures have identified regions in PrP-sen that are important in the formation of PrP-res, the exact role of PrP-sen secondary structures in the conformational transition of PrP-sen to PrP-res has not yet been defined. We constructed PrP-sen mutants with deletions of the first ß-strand, the second ß-strand, or the first a-helix and tested whether these mutants could be converted to PrP-res in both scrapie-infected neuroblastoma cells (Sc+-MNB cells) and a cell-free conversion assay. Removal of the second ß-strand or the first a-helix significantly altered both processing and the cellular localization of PrP-sen, while deletion of the first ß-strand had no effect on these events. However, all of the mutants significantly inhibited the formation of PrP-res in Sc+-MNB cells and had a greatly reduced ability to form protease-resistant PrP in a cell-free assay system. Thus, our results demonstrate that deletion of the ß-strands and the first a-helix of PrP-sen can fundamentally affect PrP-res formation and/or PrP-sen processing.
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- 2001
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12. N-terminal Truncation of Prion Protein Affects Both Formation and Conformation of Abnormal Protease-resistant Prion Protein Generatedin Vitro*
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Lawson, Victoria A., Priola, Suzette A., Wehrly, Kathy, and Chesebro, Bruce
- Abstract
Transmissible spongiform encephalopathy diseases are characterized by conversion of the normal protease-sensitive host prion protein, PrP-sen, to an abnormal protease-resistant form, PrP-res. In the current study, deletions were introduced into the flexible tail of PrP-sen (23) to determine if this region was required for formation of PrP-res in a cell-free assay. PrP-res formation was significantly reduced by deletion of residues 34–94 relative to full-length hamster PrP. Deletion of another nineteen amino acids to residue 113 further reduced the amount of PrP-res formed. Furthermore, the presence of additional proteinase K cleavage sites indicated that deletion to residue 113 generated a protease-resistant product with an altered conformation. Conversion of PrP deletion mutants was also affected by post-translational modifications to PrP-sen. Conversion of unglycosylated PrP-sen appeared to alter both the amount and the conformation of protease-resistant PrP-res produced from N-terminally truncated PrP-sen. The N-terminal region also affected the ability of hamster PrP to block mouse PrP-res formation in scrapie-infected mouse neuroblastoma cells. Thus, regions within the flexible N-terminal tail of PrP influenced interactions required for both generating and disrupting PrP-res formation.
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- 2001
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13. Efficient Conversion of Normal Prion Protein (PrP) by Abnormal Hamster PrP Is Determined by Homology at Amino Acid Residue 155
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Priola, Suzette A., Chabry, Joe¨lle, and Chan, Kaman
- Abstract
ABSTRACTIn the transmissible spongiform encephalopathies, disease is closely associated with the conversion of the normal proteinase K-sensitive host prion protein (PrP-sen) to the abnormal proteinase K-resistant form (PrP-res). Amino acid sequence homology between PrP-res and PrP-sen is important in the formation of new PrP-res and thus in the efficient transmission of infectivity across species barriers. It was previously shown that the generation of mouse PrP-res was strongly influenced by homology between PrP-sen and PrP-res at amino acid residue 138, a residue located in a region of loop structure common to PrP molecules from many different species. In order to determine if homology at residue 138 also affected the formation of PrP-res in a different animal species, we assayed the ability of hamster PrP-res to convert a panel of recombinant PrP-sen molecules to protease-resistant PrP in a cell-free conversion system. Homology at amino acid residue 138 was not critical for the formation of protease-resistant hamster PrP. Rather, homology between PrP-sen and hamster PrP-res at amino acid residue 155 determined the efficiency of formation of a protease-resistant product induced by hamster PrP-res. Structurally, residue 155 resides in a turn at the end of the first alpha helix in hamster PrP-sen; this feature is not present in mouse PrP-sen. Thus, our data suggest that PrP-res molecules isolated from scrapie-infected brains of different animal species have different PrP-sen structural requirements for the efficient formation of protease-resistant PrP.
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- 2001
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14. Species-Independent Inhibition of Abnormal Prion Protein (PrP) Formation by a Peptide Containing a Conserved PrP Sequence
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Chabry, Joe¨lle, Priola, Suzette A., Wehrly, Kathy, Nishio, Jane, Hope, James, and Chesebro, Bruce
- Abstract
ABSTRACTConversion of the normal protease-sensitive prion protein (PrP) to its abnormal protease-resistant isoform (PrP-res) is a major feature of the pathogenesis associated with transmissible spongiform encephalopathy (TSE) diseases. In previous experiments, PrP conversion was inhibited by a peptide composed of hamster PrP residues 109 to 141, suggesting that this region of the PrP molecule plays a crucial role in the conversion process. In this study, we used PrP-res derived from animals infected with two different mouse scrapie strains and one hamster scrapie strain to investigate the species specificity of these conversion reactions. Conversion of PrP was found to be completely species specific; however, despite having three amino acid differences, peptides corresponding to the hamster and mouse PrP sequences from residues 109 to 141 inhibited both the mouse and hamster PrP conversion systems equally. Furthermore, a peptide corresponding to hamster PrP residues 119 to 136, which was identical in both mouse and hamster PrP, was able to inhibit PrP-res formation in both the mouse and hamster cell-free systems as well as in scrapie-infected mouse neuroblastoma cell cultures. Because the PrP region from 119 to 136 is very conserved in most species, this peptide may have inhibitory effects on PrP conversion in a wide variety of TSE diseases.
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- 1999
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15. Neuron-specific expression of a hamster prion protein minigene in transgenic mice induces susceptibility to hamster scrapie agent
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Race, Richard E., Priola, Suzette A., Bessen, Richard A., Ernst, Darwin, Dockter, Janel, Rall, Glenn F., Mucke, Lennart, Chesebro, Bruce, and Oldstone, Michael B.A.
- Abstract
To study the effect of cell type-restricted hamster PrP expression on susceptibility to the hamster scrapie agent, we generated transgenic mice using a 1 kb hamster cDNA clone containing the 0.76 kb HPrP open reading frame under control of the neuron-specific enolase promoter. In these mice, expression of HPrP was detected only in brain tissue, with highest levels found in neurons of the cerebellu, hippocampus, thalamus, and cerebral cortex. These transgenic mice were susceptible to infection by the 263K strain of hamster scrapie with an average incubation period of 93 days, compared to 72 days in normal hamsters. In contrast, nontransgenic mice were not susceptible to this agent. These results indicate that neuron-specific expression of the 1 kb HPrP minigene including the HPrP open-reading frame is sufficient to mediate susceptibility to hamster scraple, and that HPrP expression in nonneuronal brain cells is not necessary to overcome the TSE species barrier.
- Published
- 1995
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16. A 60-kDa Prion Protein (PrP) with Properties of Both the Normal and Scrapie-associated Forms of PrP (∗)
- Author
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Priola, Suzette A., Caughey, Byron, Wehrly, Kathy, and Chesebro, Bruce
- Abstract
Scrapie is a transmissible spongiform encephalopathy of sheep and other mammals in which disease appears to be caused by the accumulation of an abnormal form of a host protein, prion protein (PrP), in the brain and other tissues. The process by which the normal protease-sensitive form of PrP is converted into the abnormal protease-resistant form is unknown. Several hypotheses predict that oligomeric forms of either the normal or abnormal PrP may act as intermediates in the conversion process. We have now identified a 60-kDa PrP derived from hamster PrP expressed in murine neuroblastoma cells. Peptide mapping studies provided evidence that the 60-kDa PrP was composed solely of PrP and, based on its molecular mass, appeared to be a PrP dimer. The 60-kDa PrP was not dissociated under several harsh denaturing conditions, which indicated that it was covalently linked. It was similar to the disease-associated form of PrP in that it formed large aggregates. However, it resembled the normal form of PrP in that it was sensitive to proteinase K and had a short metabolic half-life. The 60-kDa PrP, therefore, had characteristics of both the normal and disease-associated forms of PrP. Formation and aggregation of the 60-kDa hamster PrP occurs in uninfected mouse neuroblastoma cells, which suggests that hamster PrP has a predisposition to aggregate even in the absence of scrapie infectivity. Similar 60-kDa PrP bands were identified in scrapie-infected hamster brain but not in uninfected brain. Therefore, a 60-kDa molecule might participate in the scrapie-associated conversion of protease-sensitive PrP to protease-resistant PrP.
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- 1995
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17. Disinfection and Sterilization of Prion-Contaminated Medical Instruments
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Belay, Ermias D., Schonberger, Lawrence B., Brown, Paul, Priola, Suzette A., Chesebro, Bruce, Will, Robert G., and Asher, David M.
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- 2010
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18. Similar protein signatures for BSE and vCJD
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Priola, Suzette A.
- Published
- 1996
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19. Therapeutic Potential of Prion Protein Peptides in the Transmissible Spongiform Encephalopathies
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Priola, Suzette A.
- Published
- 2002
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