A Disulfide Intercalator Toolbox for the Site‐Directed Modification of Polypeptides

A disulfide intercalator toolbox was developed for site‐specific attachment of a broad variety of functional groups to proteins or peptides under mild, physiological conditions. The peptide hormone somatostatin (SST) served as model compound for intercalation into the available disulfide functionalization schemes starting from the intercalator or the reactive SST precursor before or after bioconjugation. A tetrazole–SST derivative was obtained that undergoes photoinduced cycloaddition in mammalian cells, which was monitored by live‐cell imaging.     

Poly(ethylene glycol)-b-poly(1,3-trimethylene carbonate) Amphiphilic Copolymers for Long-Acting Injectables: Synthesis,Non-Acylating Performance and In Vivo Degradation

This article presents the evaluation of diblock and triblock poly(ethylene glycol)-b-poly(1,3-trimethylene carbonate) amphiphilic copolymers (PEG-PTMCs) as excipients for the formulation of long-acting injectables (LAIs). Copolymers were successfully synthesised through bulk ring-opening polymerisation. The concomitant formation of PTMC homopolymer could not be avoided irrespective of the catalyst amount, but the by-product could easily be removed by gel chromatography. Pure PEG-PTMCs undergo faster erosion in vivo than their corresponding homopolymer. Furthermore, these copolymers show outstanding stability compared to their polyester analogues when formulated with amine-containing reactive drugs, which makes them particularly suitable as LAIs for the sustained release of drugs susceptible to acylation.     

Ratiometric Detection of microRNA Using Hybridization Chain Reaction and Fluorogenic Silver Nanoclusters

miRNA (miR)-155 is a potential biomarker for breast cancers. We aimed at developing a nanosensor for miR-155 detection by integrating hybridization chain reaction (HCR) and silver nanoclusters (AgNCs). HCR serves as an enzyme-free and isothermal amplification method, whereas AgNCs provide a built-in fluorogenic detection probe that could simplify the downstream analysis. The two components were integrated by adding a nucleation sequence of AgNCs to the hairpin of HCR. The working principle was based on the influence of microenvironment towards the hosted AgNCs, whereby unfolding of hairpin upon HCR has manipulated the distance between the hosted AgNCs and cytosine-rich toehold region of hairpin. As such, the dominant emission of AgNCs changed from red to yellow in the absence and presence of miR-155, enabling a ratiometric measurement of miR with high sensitivity. The limit of detection (LOD) of our HCR-AgNCs nanosensor is 1.13 fM in buffered solution. We have also tested the assay in diluted serum samples, with comparable LOD of 1.58 fM obtained. This shows the great promise of our HCR-AgNCs nanosensor for clinical application.     

Validation of a HPLC/MS method for simultaneous quantification of clonidine,morphine and its metabolites in human plasma

A high‐performance liquid chromatography–tandem mass spectrometry method was developed and validated for the simultaneous quantification of morphine, morphine's major metabolites morphine‐3‐glucuronide and morphine‐6‐glucuronide, and clonidine, to support the pharmacokinetic analysis of an ongoing double‐blinded randomized clinical trial that compares the use of morphine and clonidine in infants diagnosed with neonatal abstinence syndrome. Plasma samples were processed by solid‐phase extraction and separated on an Inertsil ODS‐3 (4 μm) column using an 0.1% formic acid in water–0.1% formic acid in methanol gradient. Detection of the analytes was conducted in the positive multiple reaction monitoring mode. The range of quantitation was 1–1000 ng/mL for morphine, morphine‐3‐glucuronide and morphine‐6‐glucuronide, and 0.25–100 ng/mL for clonidine. Intra‐day and inter‐day accuracy and precision were ≤15% for all analytes across the quantitation range. Extraction recovery rates were ≥94% for morphine, ≥90% for M3G, ≥87% for M6G and ≥ 79% for clonidine. Matrix effect ranged from 85–94% for clonidine to 101–106% for M3G. The method fulfilled all predetermined acceptance criteria and required only 100 μL of starting plasma volume. Furthermore, it was successfully applied to 30 clinical trial plasma samples.     

Binder-less and free-standing Co–Fe metal nanoparticles-decorated PVdF-HFP nanofiber membrane as an electrochemical probe for enzyme-less glucose sensors

Co–Fe bimetallic nanoparticles-affixed polyvinylidene fluoride-co-hexafluoropropylene (PVdF-HFP) nanofiber membrane is fabricated using the electrospinning and chemical reduction techniques. The semicrystalline polymeric backbone decorated with the highly crystalline Co–Fe bimetallic nanoparticles enunciates the mechanical integrity, while the incessant and swift electron mobility is articulated with the consistent dissemination of bimetallic nanoparticles on the intersected and multi-layered polymeric nanofibers. The diffusion and adsorption of glucose are expedited in the extended cavities and porosities of as-formulated polymeric nanofibers, maximizing the glucose utilization efficacy, while the uniformly implanted Co⁴⁺/Fe³⁺ active centers on PVdF-HFP nanofibers maximize the electrocatalytic activity toward glucose oxidation under alkaline regimes. Thus, the combinative sorts including nanofiber and nanocomposite strategies of PVdF-HFP/Co–Fe membrane assimilate the enzyme-less electrochemical glucose detection concerts of high sensitivity (375.01 μA mM^?1 cm^?2), low limit of detection (0.65 μm), and wide linear range (0.001 to 8 mM), outfitting the erstwhile enzyme-less glucose detection reports. Additionally, the endowments of high selectivity and real sample glucose-sensing analyses of PVdF-HFP/Co–Fe along with the binder-less and free-standing characteristics construct the state-of-the-art paradigm for the evolution of affordable enzyme-less electrochemical glucose sensors.

     

Zymography for Picogram Detection of Lipase and Esterase Activities

Lipases and esterases are important catalysts with wide varieties of industrial applications. Although many methods have been established for detecting their activities, a simple and sensitive approach for picogram detection of lipolytic enzyme quantity is still highly desirable. Here we report a lipase detection assay which is 1000-fold more sensitive than previously reported methods. Our assay enables the detection of as low as 5 pg and 180 pg of lipolytic activity by direct spotting and zymography, respectively. Furthermore, we demonstrated that the detection sensitivity was adjustable by varying the buffering capacity, which allows for screening of both high and low abundance lipolytic enzymes. Coupled with liquid chromatography-mass spectrometry, our method provides a useful tool for sensitive detection and identification of lipolytic enzymes.     

Bioprinting of Collagen: Considerations,Potentials, and Applications

Collagen is the most abundant extracellular matrix protein that is widely used in tissue engineering (TE). There is little research done on printing pure collagen. To understand the bottlenecks in printing pure collagen, it is imperative to understand collagen from a bottom‐up approach. Here it is aimed to provide a comprehensive overview of collagen printing, where collagen assembly in vivo and the various sources of collagen available for TE application are first understood. Next, the current printing technologies and strategy for printing collagen‐based materials are highlighted. Considerations and key challenges faced in collagen printing are identified. Finally, the key research areas that would enhance the functionality of printed collagen are presented.     

Regulating Oxygen Configuration in Hierarchically Porous Carbon Nanosheets for High-Rate and Durable Na+ Storage

Surface oxygen functionalities (particularly C−O configuration) in carbon materials have negative influence on their electrical conductivity and Na⁺ storage performance. Herein, we propose a concept from surface chemistry to regulate the oxygen configuration in hierarchically porous carbon nanosheets (HPCNS). It is demonstrated that the C−O/C=O ratio in HPCNS reduces from 1.49 to 0.43 and its graphitization degree increases by increasing the carbonization temperature under a reduction atmosphere. Remarkably, such high graphitization degree and low C−O content of the HPCNS-800 are favorable for promoting its electron/ion transfer kinetics, thus endowing it with high-rate (323.6 mAh g⁻¹ at 0.05 A g⁻¹ and 138.5 mAh g⁻¹ at 20.0 A g⁻¹) and durable (96 % capacity retention over 5700 cycles at 10.0 A g⁻¹) Na⁺ storage performance. This work permits the optimization of heteroatom configurations in carbon for superior Na⁺ storage.     

Recycling systems design using reservation incentive data

We consider a problem of siting compact scale biomass recycling units in residential districts, where residents can be motivated to participate in the recycling program when offered with sufficient monetary incentives. Available data samples of the residents’ reservation incentive levels are used directly to estimate the recycling participation rate in the model, which is solved for the optimal recycling unit locations and incentive-to-offer. We propose a novel approximation of the participation rate estimator that can significantly improve the tractability and scalability of the resulting mixed integer optimization model. Furthermore, we consider the effects of feedstock impurity on the economic feasibility of the recycling system, and extend our model to develop solutions that can achieve specified profit targets as well as possible in the presence of feedstock impurity. Finally, computational studies show the efficiency of the proposed model and solution approach, and give positive demonstrations of our proposed model in providing the requisite decision support in realistic recycling systems design problems.