Development of a Novel GC/MS Method for the Detection of Nicotinamide and Activity of ADP-Ribosylating Toxins

ADP-ribosylating toxin enzymes break NADH (Figure1) and transfer the ADP-ribosyl group to a residue on a target protein, permanently inactivating or denaturing the protein. This activity is typically detected with a radioassay, which is expensive and requires radioactive materials. ADP-ribosylation corresponds with the release of nicotinamide. It is possible to detect nicotinamide with a Gas Chromatograph/Mass Spectrometer (GC/MS) (Jacobson, Dame, Pyrek & Jacobson, 1995). The purpose of this study is to measure ADP-ribosylation activity using GC/MS by detecting the liberated nicotinamide. By derivatizing the nicotinamide, the detection limit was lowered to 0.5ng/μl. Control measurements of ADP-ribosylation activity by a cholera toxin protein found low levels of nicotinamide contamination

The Grizzly, November 30, 1979

College Costs To Skyrocket • Committee Explores Campus Problems • Audiovisual Services Relocated in Myrin • USGA Notes • Letters to the Editor • Roving Reporter: Iranian Crisis • Portrait of a Professor: Dr. John Wickersham • JDB - Full of Surprises • Vonnegut\u27s \u27Jailbird\u27: Life of Walter Starbuck • Pilgrim To Speak On Economy • Cagers Open 79-80 Campaign With Victory • Gridders Win Finale With Shut-Out • Garner Races To 68th In Nation • Sports Profile: Rick Morris • Pool Records Sethttps://digitalcommons.ursinus.edu/grizzlynews/1029/thumbnail.jp

Microbiomes of Thalassia testudinum throughout the Atlantic Ocean, Caribbean Sea, and Gulf of Mexico are influenced by site and region while maintaining a core microbiome

Plant microbiomes are known to serve several important functions for their host, and it is therefore important to understand their composition as well as the factors that may influence these microbial communities. The microbiome of Thalassia testudinum has only recently been explored, and studies to-date have primarily focused on characterizing the microbiome of plants in a single region. Here, we present the first characterization of the composition of the microbial communities of T. testudinum across a wide geographical range spanning three distinct regions with varying physicochemical conditions. We collected samples of leaves, roots, sediment, and water from six sites throughout the Atlantic Ocean, Caribbean Sea, and the Gulf of Mexico. We then analyzed these samples using 16S rRNA amplicon sequencing. We found that site and region can influence the microbial communities of T. testudinum, while maintaining a plant-associated core microbiome. A comprehensive comparison of available microbial community data from T. testudinum studies determined a core microbiome composed of 14 ASVs that consisted mostly of the family Rhodobacteraceae. The most abundant genera in the microbial communities included organisms with possible plant-beneficial functions, like plant-growth promoting taxa, disease suppressing taxa, and nitrogen fixers

A dual role for the N-terminal domain of the IL-3 receptor in cell signalling

The interleukin-3 (IL-3) receptor is a cell-surface heterodimer that links the haemopoietic, vascular and immune systems and is overexpressed in acute and chronic myeloid leukaemia progenitor cells. It belongs to the type I cytokine receptor family in which the α-subunits consist of two fibronectin III-like domains that bind cytokine, and a third, evolutionarily unrelated and topologically conserved, N-terminal domain (NTD) with unknown function. Here we show by crystallography that, while the NTD of IL3Rα is highly mobile in the presence of IL-3, it becomes surprisingly rigid in the presence of IL-3 K116W. Mutagenesis, biochemical and functional studies show that the NTD of IL3Rα regulates IL-3 binding and signalling and reveal an unexpected role in preventing spontaneous receptor dimerisation. Our work identifies a dual role for the NTD in this cytokine receptor family, protecting against inappropriate signalling and dynamically regulating cytokine receptor binding and function.Sophie E. Broughton, Timothy R. Hercus, Tracy L. Nero, Winnie L. Kan ... Timothy P. Hughes, Angel F. Lopez ... et al

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Development of a Novel GC/MS Method for the Detection of Nicotinamide and Activity of ADP-Ribosylating Toxins

ADP-ribosylating toxin enzymes break NADH (Figure1) and transfer the ADP-ribosyl group to a residue on a target protein, permanently inactivating or denaturing the protein. This activity is typically detected with a radioassay, which is expensive and requires radioactive materials. ADP-ribosylation corresponds with the release of nicotinamide. It is possible to detect nicotinamide with a Gas Chromatograph/Mass Spectrometer (GC/MS) (Jacobson, Dame, Pyrek & Jacobson, 1995). The purpose of this study is to measure ADP-ribosylation activity using GC/MS by detecting the liberated nicotinamide. By derivatizing the nicotinamide, the detection limit was lowered to 0.5ng/μl. Control measurements of ADP-ribosylation activity by a cholera toxin protein found low levels of nicotinamide contamination