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Andreasen posted an update 7 months, 2 weeks ago
Interest in understanding the cycling of ethanol in the environment has grown as ethanol use as a gasoline additive has increased. The production of acetaldehyde from ethanol was measured in Southern California coastal seawater. The rate of increase of acetaldehyde was positively correlated with the rate constant for ethanol biodegradation and bacteria count and was consistent with two consecutive first-order reactions where acetaldehyde is first biologically produced from ethanol then consumed. Correlation with bacteria counts suggested that acetaldehyde degradation was also a biological process. The rate constants for acetaldehyde production from ethanol and acetaldehyde loss averaged 3.0 ± 3.4 × 10-3 min-1 and 2.3 ± 4.5 × 10-2 min-1 respectively. The branching ratio for acetaldehyde production from ethanol was 0.46 ± 0.26 and estimated acetaldehyde biological production rates ranged from 0.022 to 0.800 nM min-1. With high bacterial counts, biological production rates from ethanol exceeded photochemical production rates from chromophoric dissolved organic matter. Overall, acetaldehyde production rates were larger than biodegradation rates, suggesting these waters are a source of acetaldehyde to the atmosphere. Extrapolation to higher ethanol concentrations associated with spills suggests that the production rate of acetaldehyde will initially increase and then decrease as ethanol concentrations increase.BACKGROUND AND OBJECTIVE Exenatide promotes insulin secretion and inhibits postprandial glucagon secretion. Polyethylene glycolated exenatide injection (PB-119), a derivative obtained by modification of exenatide, is more stable in metabolic behavior than exenatide in vivo. Our study aimed to evaluate the safety, tolerability and pharmacokinetic characteristics of polyethylene glycolated exenatide as a single subcutaneous injection in healthy volunteers. METHODS Seventy subjects were randomly assigned to 8 incremental dosage groups (2, 5, 10, 25, 50, 100, 200 and 400 µg). The 2- to 50-µg groups had 8 subjects in each group (the ratio of test preparation to placebo was 31), and the 100- to 400-µg groups had 10 subjects in each group (the ratio of test preparation to placebo was 41). All the subjects received a single subcutaneous injection of polyethylene glycolated exenatide and placebo according to the dosage groups. The tolerability test was conducted in the 2- to 10-µg groups. The pharmacokinetic test was tolerable for healthy volunteers. Once-weekly polyethylene glycolated exenatide injection can be recommended. Varoglutamstat datasheet CLINICAL TRIALS REGISTRATION The study was registered at clinicaltrials.gov (No. NCT02084251).Nuclear magnetic resonance (NMR) spectroscopy is a key experimental method to investigate the structure and dynamics of RNA. RNA often has only partially ordered structures responsible for its function, which makes it difficult to crystallize. In this chapter, we present the methodologies for RNA structure determination by liquid-state NMR, including the preparation of isotopically labeled RNA by in vitro transcription, NMR resonance assignment strategy, and structure calculation. Selected examples of NMR spectra are given for the first stem-loop of DsrA RNA (23 nt).Since its development, single-particle cryogenic electron microscopy (cryo-EM) has played a central role in the study at medium resolution of both bacterial and eukaryotic ribosomal complexes. With the advent of the direct electron detectors and new processing software which allow obtaining structures at atomic resolution, formerly obtained only by X-ray crystallography, cryo-EM has become the method of choice for the structural analysis of the translation machinery. In most of the cases, the ribosomal complexes at different stages of the translation process are assembled in vitro from purified components, which limit the analysis to previously well-characterized complexes with known factors composition. The initiation phase of the protein synthesis is a very dynamic process during which several proteins interact with the translation apparatus leading to the formation of a chronological series of initiation complexes (ICs). Here we describe a method to isolate ICs assembled on natural in vitro transcribed mRNA directly from rabbit reticulocyte lysate (RRL) by sucrose density gradient centrifugation . The Grad-cryo-EM approach allows investigating structures and composition of intermediate ribosomal complexes prepared in near-native condition by cryo-EM and mass spectrometry analyses. This is a powerful approach, which could be used to study translation initiation of any mRNAs, including IRES containing ones, and which could be adapted to different cell extracts.Atomic force and transmission electron microscopies (AFM/TEM) are powerful tools to analyze RNA-based nanostructures. While cryo-TEM analysis allows the determination of near-atomic resolution structures of large RNA complexes, this chapter intends to present how RNA nanostructures can be analyzed at room temperature on surfaces. Indeed, TEM and AFM analyses permit the conformation of a large population of individual molecular structures to be observed, providing a statistical basis for the variability of these nanostructures within the population. Nevertheless, if double-stranded DNA molecular imaging has been described extensively, only a few investigations of single-stranded DNA and RNA filaments have been conducted so far. Indeed, technique for spreading and adsorption of ss-molecules on AFM surfaces or TEM grids is a crucial step to avoid disturbing RNA conformation on the surface. In this chapter, we present a specific method to analyze RNA assemblies and RNA-protein complexes for molecular microscopies.Recent advances in multi-wavelength analytical ultracentrifugation (MWL-AUC) combine the power of an exquisitely sensitive hydrodynamic-based separation technique with the added dimension of spectral separation. This added dimension has opened up new doors to much improved characterization of multiple, interacting species in solution. When applied to structural investigations of RNA, MWL-AUC can precisely report on the hydrodynamic radius and the overall shape of an RNA molecule by enabling precise measurements of its sedimentation and diffusion coefficients and identify the stoichiometry of interacting components based on spectral decomposition. Information provided in this chapter will allow an investigator to design experiments for probing ion and/or protein-induced global conformational changes of an RNA molecule and exploit spectral differences between proteins and RNA to characterize their interactions in a physiological solution environment.