Non-symmetric stapling of native peptides.

Stapling has emerged as a powerful technique in peptide chemistry. It enables precise control over peptide conformation leading to enhanced properties such as improved stability and enhanced binding affinity. Although symmetric stapling methods have been extensively explored, the field of non-symmetric stapling of native peptides has received less attention, largely as a result of the formidable challenges it poses - in particular the complexities involved in achieving the high chemo-selectivity and site-selectivity required to simultaneously modify distinct proteinogenic residues. Over the past 5 years, there have been significant breakthroughs in addressing these challenges. In this Review, we describe the latest strategies for non-symmetric stapling of native peptides, elucidating the protocols, reaction mechanisms and underlying design principles. We also discuss current challenges and opportunities this field offers for future applications, such as ligand discovery and peptide-based therapeutics.

Synthesis of Glycoconjugates through Chlorooxime-Thiol Conjugation.

Introducing glycans represents an efficient chemical approach to improve the pharmacological properties of therapeutic biomolecules. Herein, we report an efficient synthesis of glycoconjugates through chlorooxime-thiol conjugation. The reactive glycosyl chlorooximes, derived from pyranoses or furanoses, readily couple to a wide range of thiol-containing substrates, including peptides, sugars, and thiophenols. This method features mild reaction conditions and fast kinetics. Capability for aqueous media and gram-scale synthesis demonstrates the potential of this method in the bioconjugation of saccharides with biologically active molecules.

3D-printed microfluidic device for high-throughput production of lipid nanoparticles incorporating SARS-CoV-2 spike protein mRNA.

Lipid nanoparticles (LNPs) are drug carriers for protecting nucleic acids for cellular delivery. The first mRNA vaccines authorized by the United States Food and Drug Administration are the mRNA-1273 (Moderna) and BNT162b (BioNTech/Pfizer) vaccines against coronavirus disease 2019 (COVID-19). We designed a 3D printed Omnidirectional Sheath-flow Enabled Microfluidics (OSEM) device for producing mRNA-loaded LNPs that closely resemble the Moderna vaccine: we used the same lipid formulations to encapsulate mRNA encoding SARS-CoV-2 spike protein. The OSEM device is made of durable methacrylate-based materials that can support flow rates in the mL min-1 range and was fabricated by stereolithography (SLA), incorporating readily adaptable interfaces using commercial fluidic connectors. Two key features of the OSEM device are: 1) a 4-way hydrodynamic flow focusing region and 2) a staggered herringbone mixer (SHM). Superior to conventional planar fluid junctions, the 4-way sheath flow channel generates an evenly focused, circular center flow that facilitates the formation of LNPs with low polydispersity. Downstream, fluid mixing in the SHM is intensified by incorporating a zig-zag fluidic pathway to deliver high mRNA encapsulation efficiency. We characterized the mRNA-loaded LNPs produced in the OSEM device and showed that the enhanced 3D microfluidic structures enable a 5-fold higher throughput production rate (60 mL min-1) of LNPs compared to commercial multi-thousand-dollar micromixers. The device produced LNPs of diameter less than 90 nm, with low polydispersity (2-8%) and high mRNA encapsulation efficiency (>90%). The 3D-printed device provides a cost-effective and easily prepared solution for high-throughput LNP production.

Recurrence and Survival Rates of Patients Who Undergo Minimally Invasive Surgery for Endometrial Carcinoma with Different Prognostic Risk Groups.

OBJECTIVE: To compare the recurrence rate and prognosis between minimally invasive surgical (MIS) approach and open surgical approach of endometrial carcinoma (EC) patients with different prognostic risk groups. METHODS: A retrospective analysis of all cases undergoing EC surgery between January 2011 and March 2018 was performed. The patients were grouped according to the management guidelines of EC patients jointly formulated by the ESGO/ESTRO/ESP 2020. Different surgical approaches were compared with regard to tumor characteristics, recurrence rate, disease-free survival (DFS), and overall survival (OS). RESULTS: A total of 665 patients met the inclusion criteria of which 196 patients underwent MIS (29.5%), and 469 patients underwent open surgery (70.5%). In the MIS group, there was a significant higher rate of recurrence (17.3% vs 6.6%, P = 0.000) compared to the open surgery group. The recurrence rate of MIS was 7.7% (P = 0.000) in the medium-high risk group and 8.2% (P = 0.014) in the high-risk group. Multivariate logistic regression analysis showed that the independent factors influencing recurrence included prognostic risk grouping, surgical approach and lymph vascular space invasion (LVSI) positivity (P < 0.05). K-M survival analysis revealed that in the intermediate and high-risk group of EC, MIS patients had a significantly lower DFS than those undergoing open surgery (P < 0.05), but no significant difference was found in OS. CONCLUSIONS: For patients with EC at moderate and high prognostic risk, MIS is associated with poorer DFS compared to open surgery, but OS was similar across prognostic risk groups. The application of MIS in patients with moderate and high-risk EC needs further research and analysis.

Polarized epithelium-sperm co-culture system reveals stimulatory factors for the secretion of mouse epididymal quiescin sulfhydryl oxidase 1.

Spermatozoa acquire fertilization ability through post-translational modifications. These membrane surface alterations occur in various segments of the epididymis. Quiescin sulfhydryl oxidases, which catalyze thiol-oxidation reactions, are involved in disulfide bond formation, which is essential for sperm maturation, upon transition and migration in the epididymis. Using castration and azoospermia transgenic mouse models, in the present study, we showed that quiescin sulfhydryl oxidase 1 (QSOX1) protein expression and secretion are positively correlated with the presence of testosterone and sperm cells. A two-dimensional in vitro epithelium-sperm co-culture system provided further evidence in support of the notion that both testosterone and its dominant metabolite, 5α-dihydrotestosterone, promote epididymal QSOX1 secretion. We also demonstrated that immature caput spermatozoa, but not mature cauda sperm cells, exhibited great potential to stimulate QSOX1 secretion in vitro, suggesting that sperm maturation is a key regulatory factor for mouse epididymal QSOX1 secretion. Proteomic analysis identified 582 secretory proteins from the co-culture supernatant, of which 258 were sperm-specific and 154 were of epididymal epithelium-origin. Gene Ontology analysis indicated that these secreted proteins exhibit functions known to facilitate sperm membrane organization, cellular activity, and sperm-egg recognition. Taken together, our data demonstrated that testosterone and sperm maturation status are key regulators of mouse epididymal QSOX1 protein expression and secretion.

Compatibility of Popular Three-Dimensional Printed Microfluidics Materials with In Vitro Enzymatic Reactions.

3D printed microfluidics offer several advantages over conventional planar microfabrication techniques including fabrication of 3D microstructures, rapid prototyping, and inertness. While 3D printed materials have been studied for their biocompatibility in cell and tissue culture applications, their compatibility for in vitro biochemistry and molecular biology has not been systematically investigated. Here, we evaluate the compatibility of several common enzymatic reactions in the context of 3D-printed microfluidics: (1) polymerase chain reaction (PCR), (2) T7 in vitro transcription, (3) mammalian in vitro translation, and (4) reverse transcription. Surprisingly, all the materials tested significantly inhibit one or more of these in vitro enzymatic reactions. Inclusion of BSA mitigates only some of these inhibitory effects. Overall, inhibition appears to be due to a combination of the surface properties of the resins as well as soluble components (leachate) originating in the matrix.

[Diversity and community characteristics of organic phosphate-mineralizing bacteria in the sea area between Minjiang Estuary to Pingtan.] / 闽江口-å¹³æ½­æµ·åŸŸæœ‰æœºè§£ç£·èŒå¤šæ ·æ€§åŠç¾¤è½ç‰¹å¾.

Microbial mineralization of organic phosphorus is an important component of marine phosphorus cycle. The research on organic phosphate-mineralizing bacteria (OPB) is helpful to reveal microbial driving mechanism of organic phosphorus mineralization in eutrophic sea area. The diversity and community characteristics of OPB were examined by Illumina high-throughput sequencing using the primer sets phoX in the sea area between Minjiang Estuary to Pingtan in April (spring) and July (summer) 2019. The results showed that the Shannon index of OPB in the surface seawater samples ranged from 3.21 to 7.91, and that the diversity at each station was greater in spring than that in summer. Shannon index of OPB in the sediment samples ranged from 2.04 to 8.70, which was greater in summer than that in spring. Shannon index of OPB in surface seawater of each station was higher than that of sediment in spring, while it was in adverse in summer. Nine phyla of OPB were detected in surface seawater, with Proteobacteria and Cyanobacteria being the most abundant. Tweleve phyla of OPB were detected in the sediments, with Proteobacteria and Bacteroidetes being the most dominant. OPB community composition at the genus level showed obvious spatio-temporal variation. Leisingera, Phaeobacter, Thalassococcus, and Pseudomonas were the major genera in the seawater in spring, while Synechococcus, Halioglobus, Roseovarius, Phaeo-bacter, Sulfitobacter, and Hyphomonas were the major genera detected in summer. Leisingera, Phaeobacter, Vibrio, and Sulfitobacter were major genera in the sediment in spring, while Azospirillum, Aminobacter, Sulfurifustis, Burkholderia, and Thiohalobacter were the major genera in summer. A large number of unclassified OPB were detected in both surface seawater and sediment. The redundancy analysis results showed that dissolved oxygen, water temperature, pH, dissolved inorganic nitrogen, NO2--N, and NO3--N had great influences on community distribution of OPB in the surface seawater. The abundant OPB in the surface seawater and sediment might play an important role in phosphorus cycle in this sea area.

Enabling Flow-Based Kinetic Off-Rate Selections Using a Microfluidic Enrichment Device.

Modern genomic sequencing efforts are identifying potential diagnostic and therapeutic targets more rapidly than existing methods can generate the peptide- and protein-based ligands required to study them. To address this problem, we have developed a microfluidic enrichment device (MFED) enabling kinetic off-rate selection without the use of exogenous competitor. We tuned the conditions of the device (bed volume, flow rate, immobilized target) such that modest, readily achievable changes in flow rates favor formation or dissociation of target-ligand complexes based on affinity. Simple kinetic equations can be used to describe the behavior of ligand binding in the MFED and the kinetic rate constants observed agree with independent measurements. We demonstrate the utility of the MFED by showing a 4-fold improvement in enrichment compared to standard selection. The MFED described here provides a route to simultaneously bias pools toward high-affinity ligands while reducing the demand for target-protein to less than a nanomole per selection.

Interfacial engineering of Ru-S-Sb/antimonene electrocatalysts for highly efficient electrolytic hydrogen generation in neutral electrolyte.

The development of high-kinetic catalysts for the hydrogen evolution reaction (HER) in a neutral electrolyte is of great importance but unfortunately remains a challenge so far. Herein, we report hybrids with abundant Ru-S-Sb bonds and engineered ultrathin antimonene (Ru-S-Sb/antimonene) as highly kinetic, active, stable electrocatalysts for the HER in an aqueous neutral electrolyte. Experiments and density functional theory (DFT) calculations reveal that Ru-S-Sb bonds coupling with antimonene synergistically work to promote HER activity. The present study brings us one step closer to understand the structure-composition-property relationships and practical electrolytic H2 production.

Liposome production and concurrent loading of drug simulants by microfluidic hydrodynamic focusing.

Liposomes are spherical vesicles enclosed by phospholipid bilayers. Nanoscale liposomes are widely employed for drug delivery in the pharmaceutical industry. In this study, nanoscale liposomes are fabricated using the microfluidic hydrodynamic focusing (MHF) approach, and the effects of flow rate ratio (FRR) on liposome size and drug loading efficiency are studied. Fluorescein isothiocyanate modified dextran is used as a hydrophilic drug simulant and Nile red is used as a hydrophobic drug simulant. The experiment results show that hydrophilic drug simulant loading efficiency increases as FRR increases and eventually plateaues at around 90% loading efficiency. The hydrophobic drug simulant loading efficiency and FRR have a positive linear correlation when FRR varies from 10 to 50. Concurrent loading of both hydrophilic and hydrophobic drug simulants maintains the same loading efficiencies as those of loading each drug simulant alone. A negative correlation between liposome size and FRR is also confirmed. Unloaded liposomes and hydrophilic drug-loaded liposomes are of the same sizes, and are smaller than the ones loaded with the hydrophobic drug simulants alone or combined. The results suggest tunable liposome size and drug loading efficiency with the MHF technique. This provides evidence to encourage further studies of microfluidic liposome fabrication in the pharmaceutical industry.

Inkjet-printed unclonable quantum dot fluorescent anti-counterfeiting labels with artificial intelligence authentication.

An ideal anti-counterfeiting technique has to be inexpensive, mass-producible, nondestructive, unclonable and convenient for authentication. Although many anti-counterfeiting technologies have been developed, very few of them fulfill all the above requirements. Here we report a non-destructive, inkjet-printable, artificial intelligence (AI)-decodable and unclonable security label. The stochastic pinning points at the three-phase contact line of the ink droplets is crucial for the successful inkjet printing of the unclonable security labels. Upon the solvent evaporation, the three-phase contact lines are pinned around the pinning points, where the quantum dots in the ink droplets deposited on, forming physically unclonable flower-like patterns. By utilizing the RGB emission quantum dots, full-color fluorescence security labels can be produced. A convenient and reliable AI-based authentication strategy is developed, allowing for the fast authentication of the covert, unclonable flower-like dot patterns with different sharpness, brightness, rotations, amplifications and the mixture of these parameters.

All-Solution-Processed Perovskite Quantum Dots Light-Emitting Diodes Based on the Solvent Engineering Strategy.

Perovskite quantum dots (PeQDs) have emerged as a new kind of nanomaterial in various applications, especially light-emitting diodes (LEDs). However, the synthesis of PeQDs is relatively complicated and the electron transport layer (ETL) is usually fabricated in a vacuum because of the dissolution of PeQDs films in organic solvents, which will increase the difficulty and cost in mass production. Here, a simple one-step "ultrasonic bath" treatment to synthesis PeQDs is adopted and applied into the PeQDs-LEDs. Meanwhile, an all-solution process is developed to fabricate PeQDs-LEDs based on the solvent engineering strategy. By using methyl acetate (MeOAc) as the solvent of ETL, the all-solution-processed PeQDs-LEDs exhibit bright luminance with the maximum current efficiency of 3.26 cd/A. This work is simple and easy to be scaled up, which will pave a new way to the low-cost all-solution processable PeQDs-LEDs.

Charging and Discharging Channels in Photoluminescence Intermittency of Single Colloidal CdSe/CdS Core/Shell Quantum Dot.

Understanding photoluminescence (PL) intermittency of single quantum dots (QDs) (intensity blinking by randomly switching between distinguishable brightness states under continuous excitation) has been a long-standing fundamental challenge and potential roadblock for their applications. Here we introduce a new analysis method for single-molecule spectroscopy that treats the blinking as photochemical/chemical processes (switching between neutral/bright and charged/dim states). It uncovers the channels for charging (bright to dim) and discharging (dim to bright) involved in PL blinking of single CdSe/CdS core/shell QDs. Both charging and discharging of the single CdSe/CdS core/shell QD possess a photochemical channel (∼10-5 to 10-6 events/photon) that linearly depends on excitation in both single- and multi-exciton regime. These two linear channels coupled to a spontaneous discharging channel (∼2 events/s) to dictate the QDs from nonblinking to gradually blinking under increasing excitation. For high-quality CdSe/CdS core/shell QDs, Auger ionization of multiexciton for both charging and discharging is negligible.

The mPEG-PCL Copolymer for Selective Fermentation of Staphylococcus lugdunensis Against Candida parapsilosis in the Human Microbiome.

Many human skin diseases, such as seborrheic dermatitis, potentially occur due to the over-growth of fungi. It remains a challenge to develop fungicides with a lower risk of generating resistant fungi and non-specifically killing commensal microbes. Our probiotic approaches using a selective fermentation initiator of skin commensal bacteria, fermentation metabolites or their derivatives provide novel therapeutics to rein in the over-growth of fungi. Staphylococcus lugdunensis (S. lugdunensis) bacteria and Candida parapsilosis (C. parapsilosis) fungi coexist in the scalp microbiome. S. lugdunensis interfered with the growth of C. parapsilosis via fermentation. A methoxy poly(ethylene glycol)-b-poly(ε-caprolactone) (mPEG-PCL) copolymer functioned as a selective fermentation initiator of S. lugdunensis, selectively triggering the S. lugdunensis fermentation to produce acetic and isovaleric acids. The acetic acid and its pro-drug diethyleneglycol diacetate (Ac-DEG-Ac) effectively suppressed the growth of C. parapsilosis in vitro and impeded the fungal expansion in the human dandruff. We demonstrate for the first time that S. lugdunensis is a skin probiotic bacterium that can exploit mPEG-PCL to yield fungicidal short-chain fatty acids (SCFAs). The concept of bacterial fermentation as a part of skin immunity to re-balance the dysbiotic microbiome warrants a novel avenue for studying the probiotic function of the skin microbiome in promoting health.

Crystal structure control of zinc-blende CdSe/CdS core/shell nanocrystals: synthesis and structure-dependent optical properties.

Nearly monodisperse zinc-blende CdSe/CdS core/shell nanocrystals were synthesized by epitaxial growth of 1-6 monolayers of CdS shell onto presynthesized zinc-blende CdSe core nanocrystals in one pot. To retain the zinc-blende structure, the reaction temperature was lowered to the 100-140 °C range by using cadmium diethyldithiocarbamate as a single-source precursor and primary amine as activation reagents for the precursor. Although the wurtzite counterparts grown under the same conditions showed optical properties similar to those reported in the literature, zinc-blende CdSe/CdS core/shell nanocrystals demonstrated surprisingly different optical properties, with ensemble single-exponential photoluminescence decay, significant decrease of photoluminescence peak width by the shell growth, and comparatively high photoluminescence quantum yields. The lifetime for the single-exponential ensemble photoluminescence decay of zinc-blende CdSe/CdS core/shell nanocrystals with 3-4 monolayers of CdS shell was reproducibly found to be approximately 16.5 ± 1.0 ns.