686 research outputs found
Purification of human butyrylcholinesterase from frozen Cohn fraction IV-4 by ion exchange and Hupresin affinity chromatography.
No full textHuman butyrylcholinesterase (HuBChE) is being developed as a therapeutic for protection from the toxicity of nerve agents. An enriched source of HuBChE is Cohn fraction IV-4 from pooled human plasma. For the past 40 years, purification of HuBChE has included affinity chromatography on procainamide-Sepharose. The present report supports a new affinity sorbent, Hupresin, for purification of HuBChE from Cohn fraction IV-4. Nine batches of 70-80 kg frozen Cohn fraction were extracted with water, filtered, and chromatographed on 30 L of Q-Ceramic ion exchange sorbent at pH 4.5. The 4% pure Q-eluent was pumped onto 4.2 L Hupresin, where contaminants were washed off with 0.3 M NaCl in 20 mM sodium phosphate pH 8.0, before 99% pure HuBChE was eluted with 0.1 M tetramethylammonium bromide. The average yield was 1.5 g of HuBChE from 80 kg Cohn paste. Recovery of HuBChE was reduced by 90% when the paste was stored at -20°C for 1 year, and reduced 100% when stored at 4°C for 24h. No reduction in HuBChE recovery occurred when paste was stored at -80°C for 3 months or 3 years. Hupresin and procainamide-Sepharose were equally effective at purifying HuBChE from Cohn fraction. HuBChE in Cohn fraction required 1000-fold purification to attain 99% purity, but 15,000-fold purification when the starting material was plasma. HuBChE (P06276) purified from Cohn fraction was a 340 kDa tetramer of 4 identical N-glycated subunits, stable for years in solution or as a lyophilized product
Solvent/detergent treated Cohn extract on 4.2 L Hupresin.
No full textSolvent/detergent treated Cohn extract on 4.2 L Hupresin.</p
ICarbS, Volume 2, Issue 1
No full textContents
General Grant and Mark Twain by John Y. Simon / 3
Ben L. Reitman, M.D.: Portrait of a Pseudo-Anarchist by Paul E. Kuhl / 11
Brian O\u27Nolan: The Privacy of His Mind by Miles Orvell / 23
Borstal Revisited by Corey Phelps / 39
James Joyce and James Stephens: The Coincidence of the Second of February by Alan M. Cohn / 61
James Joyce Collections at Morris Library by Steven P. Lund and
Alan M. Cohn / 67
Illustrations
Line drawing of Samuel Clemens by Werner Mertz / 4
Samuel Clemens to Julia Dent Grant, 3 February 1886 / 6-8
Irish Times (dummy) crowns Myles King of Ireland / 24
Manuscript of O\u27Nolan article on James Joyce / 28
Typescript of O\u27N alan light poem on Ezra Pound / 32
Manuscript opening for The Courteous Borstal / 40
Typescript from Behan\u27s Bridewell Revisited / 48
Behan typescript, Deliverance / 54
James Stephens\u27 Sarasvati / 62
Minuette, with comments by Stephens / 64
John Sullivan, James Joyce and James Stephens in Paris / 6
Human butyrylcholinesterase in Cohn fraction IV-4 purified in a single chromatography step on Hupresin.
No full textProtection from the toxicity of nerve agents is achieved by pretreatment with human butyrylcholinesterase (BChE). Current methods for purifying large quantities of BChE from frozen Cohn fraction IV-4 produce 99% pure enzyme, but the yield is low (21%). Our goal was to simplify the purification procedure and increase the yield. Butyrylcholinesterase was extracted from frozen Cohn fraction IV-4 in 10 volumes of water pH 6. The filtered extract was pumped onto a Hupresin affinity column. The previously utilized anion exchange chromatography step was omitted. Solvent and detergent reagents used to inactivate lipid enveloped virus, bacteria and protozoa did not bind to Hupresin. BChE was eluted with 0.1 M tetramethylammonium bromide in 20 mM sodium phosphate pH 8.0. BChE protein was concentrated on a Pellicon tangential flow filtration system and demonstrated to be highly purified by mass spectrometry. A high pump rate produced protein aggregates, but a low pump rate caused minimal turbidity. Possible contamination by prekallikrein and prekallikrein activator was examined by LC-MS/MS and by a chromogenic substrate assay for kallikrein activity. Prekallikrein and kallikrein were not detected by mass spectrometry in the 99% pure BChE. The chromogenic assay indicated kallikrein activity was less than 9 mU/mL. This new, 1-step chromatography protocol on Hupresin increased the yield of butyrylcholinesterase by 200%. The new method significantly reduces production costs by optimizing yield of 99% pure butyrylcholinesterase
Did the 2007 Legal Arizona Workers Act Reduce the State's Unauthorized Immigrant Population?
No full textWe test for an effect of Arizona’s 2007 Legal Arizona Workers Act (LAWA) on the proportion of the state population characterized as foreign-born, as non-citizen, and as non-citizen Hispanic. We use the synthetic control method to select a group of states against which the population trends of Arizona can be compared. We document a notable and statistically significant reduction in the proportion of the Arizona population that is foreign-born and in particular, that is Hispanic noncitizen. The decline observed for Arizona matches the timing of LAWA's implementation, deviates from the time series for the chosen synthetic control group, and stands out relative to the distribution of placebo estimates for the remainder of states in the nation. Furthermore, we do not observe similar declines for Hispanic naturalized citizens, a group not targeted by the legislation. Our results on LAWA's impact on the housing market provide further support for our findings.illegal, unauthorized, undocumented, immigration, Hispanic, Arizona
Comparison of BChE yield with and without solvent/detergent (S/D) in the Cohn extract.
No full textComparison of BChE yield with and without solvent/detergent (S/D) in the Cohn extract.</p
Summary of HuBChE yield from Cohn paste extract purified on 30 L of Q-Ceramic<sup>a</sup>.
No full textSummary of HuBChE yield from Cohn paste extract purified on 30 L of Q-Ceramica.</p
Writing and the rights of reality: usurpation and potentiality in Derrida, Plato, Nietzsche, and Beckett
No full textThe thesis critically evaluates Jacques Derrida's conferral of the rights of reality on writing, focussing on his theory of an arche-text in light of the speculative nature of this theory. The theory is initially considered in the context of Derrida's elucidation of the usurpatory status of writing within the Platonic and Nietzschean texts. This consideration reveals an admission of writing's usurpatory status by both writers while at the same time demonstrating their awareness of the intrinsically speculative nature of this view, the significance of writing lying in its ability to exteriorise the radically indeterminate status of consciousness m relation to reality rather than its ability to displace consciousness or reality The analyses, therefore, not only bring the Derridean hypothesis of a repressive or phonocentric metaphysical episteme into question but also exhibit the historical and philosophical role of potentiality in relation to writing, writing's ultimate significance lying in its capacity to exteriorise our existence as a mode of potentiality. Accordingly, in the second half of the thesis the Derridean theory of writing is countered with a specifically Aristotelian theory of the text as it is exhibited in the prose of Samuel Beckett, an author whose significance lies in his close alignment with Derridean theory within contemporary criticism. It is demonstrated that this identification has obviated an awareness of the significance of potentiality within the Beckettian text, his work consequently being appraised in the previously neglected context of Aristotelian metaphysics
2005 Bill Summary
No full textMEMBERS: Mark Lena, Chair Jay La Suer, Vice Chair Rebecca Cohn Mervyn M. Dymally Jackie Goldberg Todd Spitzer Vacancy
COMMITTEE STAFF Gregory Pagan, Chief Counsel Kathleen Ragan, Counsel Heather Hopkins, Counsel Steven Meinrath, Counsel Kimberly Horiuchi, Counsel
Sue Highland, Committee Secretary Toni J. Nakashima, Committee Secretar
Purification of human butyrylcholinesterase from frozen Cohn fraction IV-4 by ion exchange and Hupresin affinity chromatography - Fig 9
No full textNondenaturing polyacrylamide gradient gel stained for BChE activity (A) and counterstained with Coomassie blue (B). HuBChE eluted from Hupresin (lanes 1–8) consists predominantly of tetramers (C4). Desialylated HuBChE in the Cohn extract and the Q-Ceramic pass-thru migrates near the top of the gel in a broad band (lanes 9 and 10). Cohn paste extract (lane 9) contains partly desialylated HuBChE. The desialylated HuBChE did not bind to Q-Ceramic at pH 4.5, but passed through as visualized by the intense staining for the Q-Ceramic pass through (lane 10). Faint bands of activity are best seen in panel A where C1 is the HuBChE monomer, C2 is an albumin-HuBChE dimer [25], and C3 is a HuBChE dimer. The major band of HuBChE activity is the tetramer C4. The gel counterstained with Coomassie blue (panel B) shows the same pattern of blue bands in lanes 9 and 10. This means the major protein contaminants in the Cohn extract (lane 9) pass through Q-Ceramic without binding (lane 10).</p
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