Analysis of Protein Turnover by Quantitative SNAP-Based Pulse-Chase Imaging

Assessment of protein dynamics in living cells is crucial for understanding their biological properties and functions. The SNAP-tag, a self labeling suicide enzyme, presents a tool with unique features that can be adopted for determining protein dynamics in living cells. Here we present detailed protocols for the use of SNAP in fluorescent pulse-chase and quench-chase-pulse experiments. These time-slicing methods provide powerful tools to assay and quantify the fate and turnover rate of proteins of different ages. We cover advantages and pitfalls of SNAP-tagging in fixed- and live-cell studies and evaluate the recently developed fast-acting SNAPf variant. In addition, to facilitate the analysis of protein turnover datasets, we present an automated algorithm for spot recognition and quantification.

A Comprehensive Discussion of HMBC Pulse Sequences: 4. Establishing Two-Bond Correlations from HMBC and Related Experiments

The utility of the HMBC experiment for structure elucidation is unquestionable, but the nature of the coupling pathways leading to correlations in an HMBC experiment creates the potential for misinterpretation. This misinterpretation potential is intimately linked to the size of the long-range heteronuclear couplings involved, and may become troublesome in those cases of a particularly strong 2JCH correlation that might be mistaken for a 3JCH correlation or a 4JCH correlation of appreciable strength that could be mistaken for a weaker 3JCH correlation. To address these potential avenues of confusion, work from several laboratories has been focused on the development of what might be considered “coupling pathway edited” long-range heteronuclear correlation experiments that are derived from or related to the HMBC experiment. The first example of an effort to address the problems associated with correlation path length was seen in the heteronucleus-detected XCORFE experiment described by Reynolds and co-workers that predated the development of the HMBC experiment. Proton-detected analogs of the HMBC experiment intended to differentiate 2JCH correlations from nJCH correlations where n = 3, 4, include the 2J,3J-HMBC, HMBC-RELAY, H2BC, edited-HMBC, and HAT H2BC experiments. The principles underlying the critical components of each of these experiments are discussed and experimental verification of the results that can be obtained using model compounds are shown. This contribution concludes with a brief discussion of the 1,1-ADEQUATE experiments that provide an alternative means of identifying adjacent protonated and non-protonated carbon correlations by exploiting 1JCC correlations at natural abundance.

(Table 2) Delta 14C record of sediment core Pulse-32_SMB

The D14C of surface water dissolved inorganic carbon (DIC) in the Southern California Bight was compared to D14C as recorded by the sterols in Santa Monica and Santa Barbara Basin sediments. All of the C26, C27, C28, and C29 sterols as well as dinosterol had 14C concentrations equal to surface water DIC, indicating that all of the major sterols were derived from phytoplanktonic production. There is no detectable terrestrial component. Their tracer capability was confirmed by comparing the "bomb 14C"-derived change in surface water D14CDIC with the change in D14Csterol. The "prebomb" D14CDIC was -82 per mil, and prebomb sterols averaged -75±19 per mil. The D14C value in 1996 was +71 per mil. Eighteen measurements representing eight different sterols from the sediment-water interface of both Santa Monica and Santa Barbara Basins averaged +62±23 per mil. When three of these values were eliminated because of suspected contamination, the remaining data averaged +71 ±12 per mil. The entire compound class could serve as an excellent proxy for the 14C concentration of ocean surface waters.

Micromesistius poutassou (blue whiting) abundance and biomass data in Bay of Biscay estimated from acoustic surveys (2000-2012)

Acoustic and pelagic trawl data were collected during various pelagic surveys carried out by IFREMER in May between 2000 and 2012 (except 2001), on the eastern continental shelf of the Bay of Biscay (Pelgas series). The acoustic data were collected with a Simrad EK60 echosounder operating at 38 kHz (beam angle at -3 dB: 7°, pulse length set to 1.024 ms). The echosounder transducer was mounted on the vessel keel, at 6 m below the sea surface. The sampling design were parallel transects spaced 12 nm apart which were orientated perpendicular to the coast line from 20 m to about 200 m bottom depth. The nominal sailing speed was 10 knots and 3 knots on average during fishing operations. The scrutinising (species identification) of acoustic data was done by first characterising acoustic schools by type and then linking these types with the species composition of specific trawl hauls. The data set contains nautical area backscattering values, biomass and abundance estimates for blue whiting for one nautical mile long transect lines. Further information on the survey design, scrutinising and biomass estimation can be found in Doray et al. 2012.

Fast neutron pulse reactors and fast neutron critical assemblies : a literature search /

"Reactor Technology."