In Vitro Assay and Characterization of the Farnesylation-Dependent Prelamin a Endoprotease

The 72-kDa nuclear lamina protein lamin A is synthesized as a 74-kDa farnesylated precursor. Conversion of this precursor to mature lamin A appears to be mediated by a specific endoprotease. Prior studies of overexpressed wild-type and mutant lamin A proteins in cultured cells have indicated that the precursor possesses the typical carboxyl-terminal S- farnesylated, cysteine methyl ester and that farnesylation is required for endoproteolysis to occur. In this report, we describe the synthesis of an S- farnesyl, cysteinyl methyl ester peptide corresponding to the carboxyl- terminal 18 amino acid residues of human prelamin A. This peptide acts as a substrate for the prelamin A endoprotease in vitro, with cleavage of the synthetic peptide at the expected site between Tyr657 and Leu658. Endoproteolytic cleavage requires the S-prenylated cysteine methyl ester and, in agreement with transfection studies, is more active with the farnesylated than geranylgeranylated cysteinyl substrate. N-Acetyl farnesyl methyl cysteine is shown to be a noncompetitive inhibitor of the enzyme. Taken together, these observations suggest that there is a specific farnesyl binding site on the enzyme which is not at the active site

Imprints, Vol. 2

Imprints is the official publication for Sigma Tau Delta, the honorary English fraternity. The editors welcome creative works submitted by contributors and also publish winners of the annual T. E. Ferguson Writing Contest. Especially welcome are poems, fiction pieces and essays of no more than 5,000 words in length. At this time we would like to express our gratitude to David Whitescarver, Sigma Tau Delta faculty advisor, for his unrelenting optimism and valuable help in the preparation of this journal

Haunted by the Presence of Death: Prisons, Abolitionism and the Right to Life

This chapter explores how prisons in England and Wales are haunted by the presence of death. It details how prisoners experience civil death (death in law), social death (death as a worthy human being) and corporeal death (literal death of the body). The chapter discusses two different but associated abolitionist strategies to contest the prison as a place of death: (i) naming the people who have died and recognising their continued humanity, as a way to promote greater penal accountability; and, (ii) direct action as a way of ‘making something happen’. Overall, the chapter points to the need for a dedicated democratic public space (an agora) committed to rational, informed debate that recognises the inherent deadly outcomes of imprisonment

A measure of centrality based on the spectrum of the Laplacian

We introduce a family of new centralities, the k-spectral centralities. k-Spectral centrality is a measurement of importance with respect to the deformation of the graph Laplacian associated with the graph. Due to this connection, k-spectral centralities have various interpretations in terms of spectrally determined information. We explore this centrality in the context of several examples. While for sparse unweighted networks 1-spectral centrality behaves similarly to other standard centralities, for dense weighted networks they show different properties. In summary, the k-spectral centralities provide a novel and useful measurement of relevance (for single network elements as well as whole subnetworks) distinct from other known measures.Comment: 12 pages, 6 figures, 2 table

Split-time AI : using estrus detection aids to optimize timed artificial insemination (2016)

New July 2016.Includes bibliographical reference

Facing the River Gauntlet: Understanding the Effects of Fisheries Capture and Water Temperature on the Physiology of Coho Salmon

An improved understanding of bycatch mortality can be achieved by complementing field studies with laboratory experiments that use physiological assessments. This study examined the effects of water temperature and the duration of net entanglement on physiological disturbance and recovery in coho salmon (Oncorhynchus kisutch) after release from a simulated beach seine capture. Heart rate was monitored using implanted electrocardiogram biologgers that allowed fish to swim freely before and after release. A subset of fish was recovered in respirometers to monitor metabolic recovery, and separate groups of fish were sacrificed at different times to assess blood and white muscle biochemistry. One hour after release, fish had elevated lactate in muscle and blood plasma, depleted tissue energy stores, and altered osmoregulatory status, particularly in warmer (15 vs. 10°C) and longer (15 vs. 2 min) capture treatments. A significant effect of entanglement duration on blood and muscle metabolites remained after 4 h. Oxygen consumption rate recovered to baseline within 7–10 h. However, recovery of heart rate to routine levels was longer and more variable, with most fish taking over 10 h, and 33% of fish failing to recover within 24 h. There were no significant treatment effects on either oxygen consumption or heart rate recovery. Our results indicate that fishers should minimize handling time for bycatch and maximize oxygen supply during crowding, especially when temperatures are elevated. Physiological data, such as those presented here, can be used to understand mechanisms that underlie bycatch impairment and mortality, and thus inform best practices that ensure the welfare and conservation of affected species

Split-time Al : using estrus detection aids to optimize timed artificial insemination (2018)

Timed AI pregnancy rates can be optimized through use of a split-time AI approach following administration of the 14-day CIDR-PG protocol for heifers and the 7-day COSynch + CIDR protocol for mature cows. Using split-time AI, insemination of non-estrous females is delayed until 20 to 24 hours after the scheduled time for fixed-time AI. Estrotect estrus detection aids applied at the time of PG administration allow producers to determine the estrous status of females and inseminate at the optimal time.Includes bibliographical reference

A comparability study of 5 commercial KRAS tests

<p>Abstract</p> <p>Background</p> <p>Activating mutations in the <it>KRAS </it>gene occur frequently in human tumors, including colorectal carcinomas; most mutations occur in codons 12 and 13. Mutations in <it>KRAS </it>have been associated with poor response to anti-epidermal growth factor receptor antibodies. Therefore, an accurate and readily available analysis of <it>KRAS </it>mutational status is needed. The aim of this study was to evaluate concordance between <it>KRAS </it>assays performed by 6 different laboratories.</p> <p>Methods</p> <p>Forty formalin-fixed paraffin-embedded colorectal cancer tumor samples were obtained. Sample sections were submitted for <it>KRAS </it>mutation analysis to 5 independent commercial laboratories (Agencourt, Gentris, Genzyme, HistoGeneX, and Invitek) and to the Amgen DNA Sequencing Laboratory for direct polymerase chain reaction sequencing. The assay used by Invitek is no longer commercially available and has been replaced by an alternative technique. Results from the commercial services were compared with those from Amgen direct sequencing by κ statistics.</p> <p>Results</p> <p><it>KRAS </it>mutations were observed in codon 12 and/or 13 in 20 of 40 (50%) samples in Amgen direct sequencing assays. Results from HistoGeneX (κ = 0.95), Genzyme (κ = 0.94), and Agencourt (κ = 0.94) were in almost perfect agreement with these results, and the results from Gentris were in substantial agreement with the results from Amgen (κ = 0.75). The Invitek allele-specific assay demonstrated slight agreement (κ = 0.13).</p> <p>Conclusions</p> <p>This study provides data on the comparability of <it>KRAS </it>mutational analyses. The results suggest that most (but not all) commercial services provide analysis that is accurate and comparable with direct sequencing.</p

Brief for the United States as Amicus Curiae in Support of Neither Party

Amicus ("friend of the court") brief written by the United States in support of petitioners in AMP v. Myriad Genetics (Supreme Court Case Docket No. 12-398)