Activity

Synthetic peptides are a critical requirement for the development and application of targeted mass spectrometry (MS)-based assays for the quantitation of proteins from biological matrices. Y-27632 Transporting synthetic peptides on dry ice from one laboratory to another is costly and often difficult because of country-specific import and export regulations. Therefore, in this study, we assessed the impact of leaving a lyophilized mixture consisting of 125 peptides at room temperature for up to 20 days, and we assessed the effect on the quantitative performance of multiple reaction monitoring-MS (MRM-MS) assays. The findings suggest that there are no significant differences in the MRM-MS results for the time points assessed in this study (up to 20 days). All the calibration curves and quality control (QC) samples met the acceptance criteria for precision and accuracy (raw data are available via the public MS data repository PanoramaWeb, identifier /MRM Proteomics/2020_BAK125_RT). The number of endogenous proteins quantifiable across five plasma samples was consistently between 87 and 99 out of 125 for all time points. Moreover, the coefficients of variation (CVs) calculated for the majority of peptide concentrations across all samples and time points were less then 5%. In addition, a lyophilized peptide mixture was transported from Canada to Iceland without dry ice. The results showed that there was no significant difference in the quantitative performance, with the determined concentrations of most proteins in the samples falling within 30% between the analyses performed on the same three plasma samples in Iceland and those in Canada. Overall, a comparison of the results obtained in Canada and in Iceland indicated that the peptides were stable under the conditions tested and also indicated that shipping lyophilized peptide mixtures without dry ice, but in the presence of sufficient desiccant material, could be a feasible option in cases where transport difficulties may arise or dry-ice sublimation may occur.Alkyl and aryl halides have been studied extensively as radical precursors; however, mild and less toxic conditions for the activation of alkyl bromides toward alkyl radicals are still desirable. Reported here is a reductive radical conjugate addition that allows for the formation of alkyl radicals via activation of alkyl bromides through cobalt/iridium catalysis. The developed conditions are emphasized in the broad substrate scope presented, including benzylic halides and halides containing free alcohols, silanes, and chlorides.A kind of blocked aptamer-functionalized molecular beacon (MB) was designed as fluorescence sensors to detect thrombins by binding-induced “turn on” structural transformation. Three MBs named MB(8 + 8), MB(15 + 8), and MB(15 + 6) consisted of two single-stranded oligonucleotides. One long single-stranded oligonucleotide (abbreviated as SS) contained a thrombin aptamer sequence and was modified with a fluorescence group and quenching group on each end side. Another short single-stranded oligonucleotide (written as cDNA) was partially complementary to the long SS. It was interesting to find that the complementary sequence length of cDNA greatly influenced the structure of the MBs. The construction of MB experiments proved that MB(8 + 8) and MB(15 + 8) could form the quenching MBs but MB(15 + 6) could not. MB(8 + 8) was composed of a SS strand paired with a complementary cDNA(8 + 8), which was called one-to-one combination, while MB(15 + 8) was two-to-two combination and MB(15 + 6) was one-to-two combination. When the ratio of SS and cDNA (15 + 8) was 11, the quenching efficiency reached maximum. But with the molar ratio of SS and cDNA(8 + 8) increasing, the quenching efficiency increased continuously. Under the optimal conditions that we studied, the detection limit of thrombin by MB(8 + 8) and MB(15 + 8) was 0.19 and 1.2 nM, respectively. In addition, the assay proved to be selective, and the average recovery of thrombin detected by MB(8 + 8) and MB(15 + 8) in diluted serum was 95.4 and 94.5%, respectively.The controllable integration of low-dimensional nanomaterials on solid surfaces is pivotal for the fabrication of next-generation miniaturized electronic and optoelectronic devices. For instance, organization of two-dimensional (2D) nanomaterials on polymeric surfaces paves the way for the development of flexible electronics for applications in wearable devices. Nevertheless, the understanding of the molecular interactions between these nanomaterials and the polymeric surfaces remains limited, which impedes the rational design of 2D nanomaterial-based functional coatings. In the current work, we report that graphene oxide (GO) nanosheets, in their dispersion phase, can be adsorbed on multiple polymeric surfaces in a spontaneous manner. Both experimental findings and simulational results indicate that the main driving force is hydrogen bonding interactions, although other molecular interactions such as polarity and dispersion ones contribute to the adsorption as well. The relatively high hydrogen bonding interactions cause not only increased GO surface coverage but also enhanced GO adsorption kinetics on polymeric surfaces. The adsorbed GO layers are robust, which can be explained by the large aspect ratios of GO nanosheets and the presence of multiple spots for molecular interactions. As a proof of concept, GO-covered polymethyl methacrylate effectively decreases surface static charges when compared with its pristine counterpart. The integration of the GO constituents turns many inert polymeric substrates into multifunctional hybrids, and the functional groups on GO can be used further to bridge with additional functional materials for the development of high-performance electronic devices.We present novel data on the composition-, pH-, and salt-dependent zero shear viscosity of the commercially important mixture of anionic sodium dodecyl sulfate (SDS) and zwitterionic lauramidopropyl betaine (LAPB). We show via proton NMR experiments that the notionally zwitterionic LAPB exhibits a large pKa shift in the presence of SDS and can become partially cationic at formulation-relevant pH ranges of 4.5-6.0-that is, the binary system is effectively a ternary system. This has a pronounced effect on the viscosity of the system at low pH, especially if the fraction of LAPB is high. We use theoretical arguments to motivate a semiempirical but practical approach to model the viscosity of the mixtures using thermodynamic parameters such as the excess chemical potentials or activity coefficients of the surfactants. We demonstrate this using an augmented regular solution theory-based mixed micelle thermodynamic model and develop robust regression models using Bayesian approaches. We also show how the pKa shift from NMR experiments can be used to parameterize the thermodynamic model. This framework should be extensible to other arbitrary surfactant mixtures in the future and hence will be of broad interest for the development of surfactant formulations for household, personal care, and other applications.Protected dipeptides can be converted into cyclic ketoaminals, which can be subjected to palladium-catalyzed regioselective C-H functionalization. The best results are obtained using the 2-(methylthio)aniline (MTA) directing group, which is superior to the commonly used 8-aminoquinoline (AQ) group. No epimerization of stereogenic centers is observed. Subsequent cleavage of the directing and protecting groups allows the incorporation of a modified dipeptide into larger peptide chains.Once protein synthesis is excessive or misfolded protein becomes aggregated, which eventually overwhelms the capacity of the endoplasmic reticulum (ER), a state named ER stress would be reached. ER stress could affect many tissues, especially the liver, in which nonalcoholic fatty liver disease, liver steatosis, etc. have been reported relative. However, there is still a lack of systematic insight into ER stress in the liver, which can be obtained by integrating metabolomics and transcriptomics of the tissue. Here, tunicamycin was utilized to induce ER stress in C57BL/6N mice. Microarray and untargeted metabolomics were performed to identify the genes and metabolites significantly altered in liver tissues. Surprisingly, apart from the predictable unfolded protein response, liver lipid, arginine, and proline metabolisms were affirmed to be related to ER stress. Also, the ketone body metabolism changed most prominently in response to ER stress, with few studies backing. What is more, succinate receptor 1 (Sucnr1) may be a novel marker and therapeutical target of liver ER stress. In this study, the combination of the metabolome and transcriptome provided reliable information about liver pathological processes, including key relative pathways, potential markers, and targets involved in ER stress of the liver.The ability to control the motion of single nanoparticles or molecules is currently one of the major scientific and technological challenges. Despite tremendous progress in the field of plasmonic nanotweezers, controlled nanoscale manipulation of nanoparticles trapped by a plasmonic nanogap antenna has not been reported yet. Here, we demonstrate the controlled orbital rotation of a single fluorescent nanodiamond trapped by a gold trimer nanoantenna irradiated by a rotating linearly polarized light or circularly polarized light. Remarkably, the rotation direction is opposite to the light’s polarization rotation. We numerically show that this inversion comes from sequential excitation of individual nanotriangles in the reverse order when the linear polarization is rotated, whereas using a circular polarization, light-nanoparticle angular momentum transfer occurs via the generation of a Poynting vector vortex of reversed handedness. This work provides a new path for the control of light-matter angular momentum transfer using plasmonic nanogap antennas.Anabaena sp. PCC 7120 (Anabaena 7120) is a photoautotrophic filamentous cyanobacterium capable of fixing atmospheric nitrogen. It is a model organism used for studying cell differentiation and nitrogen fixation. Under nitrogen deficiency, Anabaena 7120 forms specialized heterocysts capable of nitrogen fixation. However, the molecular mechanisms involved in the cyanobacterial adaptation to nitrogen deficiency are not well understood. Here, we employed a label-free quantitative proteomic strategy to systematically investigate the nitrogen deficiency response of Anabaena 7120 at different time points. In total, 363, 603, and 669 proteins showed significant changes in protein abundance under nitrogen deficiency for 3, 12, and 24 h, respectively. With mapping onto metabolic pathways, we revealed proteomic perturbation and regulation of carbon and nitrogen metabolism in response to nitrogen deficiency. Functional analysis confirmed the involvement of nitrogen stress-responsive proteins in biological processes, including nitrogen fixation, photosynthesis, energy and carbon metabolism, and heterocyst development. The expression of 10 proteins at different time points was further validated by using multiple reaction monitoring assays. In particular, many dysregulated proteins were found to be time-specific and involved in heterocyst development, providing new candidates for future functional studies in this model cyanobacterium. These results provide novel insights into the molecular mechanisms of nitrogen stress responses and heterocyst development in Anabaena 7120.