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SUMMARY Both hospital- and community-acquired Staphylococcus aureus infections have become major health concerns in terms of morbidity, suffering and cost. Trimethoprim-sulfamethoxazole (TMP-SMZ) is an alternative treatment for methicillin-resistant S. aureus (MRSA) infections. However, TMP-resistant strains have arisen with point mutations in dihydrofolate reductase (DHFR), the target for TMP. A single point mutation, F98Y, has been shown biochemically to confer the majority of this resistance to TMP. Using a structure-based approach, we have designed a series of novel propargyl-linked DHFR inhibitors that are active against several trimethoprim-resistant enzymes. We screened this series against wild-type and mutant (F98Y) S. aureus DHFR and found that several are active against both enzymes and specifically that the meta-biphenyl class of these inhibitors is the most potent. In order to understand the structural basis of this potency, we determined eight high-resolution crystal structures: four each of the wild-type and mutant DHFR enzymes bound to various propargyl-linked DHFR inhibitors. In addition to explaining the structure-activity relationships, several of the structures reveal a novel conformation for the cofactor, NADPH. In this new conformation that is predominantly associated with the mutant enzyme, the nicotinamide ring is displaced from its conserved location and three water molecules complete a network of hydrogen bonds between the nicotinamide ring and the protein. In this new position, NADPH has reduced interactions with the inhibitor. An equilibrium between the two conformations of NADPH, implied by their occupancies in the eight crystal structures, is influenced both by the ligand and the F98Y mutation. The mutation induced equilibrium between two NADPH binding conformations may contribute to decrease TMP binding and thus may be responsible for TMP resistance.

Background The therapeutic potential of exosomes derived from stem cells has attracted increasing interest recently, because they can exert similar paracrine functions of stem cells and overcome the limitations of stem cells transplantation. Exosomes derived from bone mesenchymal stem cells (BMSC-Exos) have been confirmed to promote osteogenesis and angiogenesis. The magnetic nanoparticles (eg. Fe3O4, γ-Fe2O3) combined with a static magnetic field (SMF) has been commonly used to increase wound healing and bone regeneration. Hence, this study aims to evaluate whether exosomes derived from BMSCs preconditioned with a low dose of Fe3O4 nanoparticles with or without the SMF, exert superior pro-osteogenic and pro-angiogenic activities in bone regeneration and the underlying mechanisms involved. Methods Two novel types of exosomes derived from preconditioned BMSCs that fabricated by regulating the contents with the stimulation of magnetic nanoparticles and/or a SMF. Then, the new exosomes were isolated by ultracentrifugation and characterized. Afterwards, we conducted in vitro experiments in which we measured osteogenic differentiation, cell proliferation, cell migration, and tube formation, then established an in vivo critical-sized calvarial defect rat model. The miRNA expression profiles were compared among the exosomes to detect the potential mechanism of improving osteogenesis and angiogenesis. At last, the function of exosomal miRNA during bone regeneration was confirmed by utilizing a series of gain- and loss-of-function experiments in vitro. Results 50 µg/mL Fe3O4 nanoparticles and a 100 mT SMF were chosen as the optimum magnetic conditions to fabricate two new exosomes, named BMSC-Fe3O4-Exos and BMSC-Fe3O4-SMF-Exos. They were both confirmed to enhance osteogenesis and angiogenesis in vitro and in vivo compared with BMSC-Exos, and BMSC-Fe3O4-SMF-Exos had the most marked effect. The promotion effect was found to be related to the highly riched miR-1260a in BMSC-Fe3O4-SMF-Exos. Furthermore, miR-1260a was verified to enhance osteogenesis and angiogenesis through inhibition of HDAC7 and COL4A2, respectively. Conclusion These results suggest that low doses of Fe3O4 nanoparticles combined with a SMF trigger exosomes to exert enhanced osteogenesis and angiogenesis and that targeting of HDAC7 and COL4A2 by exosomal miR-1260a plays a crucial role in this process. This work could provide a new protocol to promote bone regeneration for tissue engineering in the future. Graphical abstract

High-performance artificial synaptic devices are indispensable for developing neuromorphic computing systems with high energy efficiency. However, the reliability and variability issues of existing devices such as nonlinear and asymmetric weight update are the major hurdles in their practical applications for energy-efficient neuromorphic computing. Here, a two-terminal floating-gate memory (2TFGM) based artificial synapse built from all-2D van der Waals materials is reported. The 2TFGM synaptic device exhibits excellent linear and symmetric weight update characteristics with high reliability and tunability. In particular, the high linearity and symmetric synaptic weight realized by simple programming with identical pulses can eliminate the additional latency and power consumption caused by the peripheral circuit design and achieve an ultralow energy consumption for the synapses in the neural network implementation. A large number of states up to ≈3000, high switching speed of 40 ns and low energy consumption of 18 fJ for a single pulse have been demonstrated experimentally. A high classification accuracy up to 97.7% (close to the software baseline of 98%) has been achieved in the Modified National Institute of Standards and Technology (MNIST) simulations based on the experimental data. These results demonstrate the potential of all-2D 2TFGM for high-speed and low-power neuromorphic computing.

modulate their electrical, optical, and structural properties by introducing impurities for doping. [5-10] So far, the chemisorption and charge transfer through surface functionalization are effective approaches to achieve doping, but it has a relatively weak influences on band structures or improvements of electronic properties. [11,12] By contrast, substitutional doping is more stable and capable of tailoring the bandgap of 2D-TMDs without introducing in-gap states. [13-16] Such substitutional doping could be realized through replacing the host transition metal or chalcogen atoms with other elements during synthesis to form alloys such as Mo 1-x W x S 2 , MoS 2x Se 2(1-x) , and V x W y Mo 1-x-y S 2z Se 2(1-z) , etc. [17-21] Recently, oxygen doping of 2D-MoS 2 has attracted considerable research interests. Previous studies show that post treatments of intrinsic MoS 2 in air, ozone, or oxygen plasma could induce oxygen doping, evidenced from the enhanced catalytic reactivity, quenched photoluminescence, and improved electrical conductivity. [9,18,22-24] Such doping processes usually lead to oxygen substitution and oxidation as well and consequently yield highly disordered or fragmented lattice structures. In situ oxygen substitutional doping in 2D-MoS 2 with a controlled and nondestructive manner is thus highly desired to preserve its original lattice structure, but still remains challenging so far. In 2D semiconductors, doping offers an effective approach to modulate their optical and electronic properties. Here, an in situ doping of oxygen atoms in monolayer molybdenum disulfide (MoS 2) is reported during the chemical vapor deposition process. Oxygen concentrations up to 20-25% can be reliable achieved in these doped monolayers, MoS 2-x O x. These oxygen dopants are in a form of substitution of sulfur atoms in the MoS 2 lattice and can reduce the bandgap of intrinsic MoS 2 without introducing in-gap states as confirmed by photoluminescence spectroscopy and scanning tunneling spectroscopy. Field effect transistors made of monolayer MoS 2-x O x show enhanced electrical performances, such as high field-effect mobility (≈100 cm 2 V-1 s-1) and inverter gain, ultrahigh devices' on/off ratio (>10 9) and small subthreshold swing value (≈80 mV dec-1). This in situ oxygen doping technique holds great promise on developing advanced electronics based on 2D semiconductors.

Background: Both magnetic nanoparticles (MNPs) and exosomes derived from bone mesenchymal stem cells (BMSC-Exos) have been reported to improve wound healing. In this study, novel exosomes (mag-BMSC-Exos) would be fabricated from BMSCs with the stimulation of MNPs and a static magnetic field (SMF) to further enhance wound repair. Methods: Mag-BMSC-Exos, namely, exosomes derived from BMSCs preconditioned with Fe 3 O 4 nanoparticles and a SMF, together with BMSC-Exos were both first isolated by ultracentrifugation, respectively. Afterwards, we conducted in vitro experiments, including scratch wound assays, transwell assays, and tube formation assays, and established an in vivo wound healing model. The miRNA expression profiles were compared between BMSC-Exos and mag-BMSC-Exos to detect the potential mechanism of improving wound healing. At last, the function of exosomal miR-21-5p during wound healing was confirmed by utilizing a series of gain-and loss-of-function experiments in vitro. Results: The optimal working magnetic condition was 50 µg/mL Fe 3 O 4 nanoparticles combined with 100 mT SMF. In vitro, mag-BMSC-Exo administration promoted proliferation, migration and angiogenesis to a greater extent than BMSC-Exo administration. Local transplantation of mag-BMSC-Exos into rat skin wounds resulted in accelerated wound closure, narrower scar widths and enhanced angiogenesis compared with BMSC-Exo transplantation. Notably, miR-21-5p was found to be highly enriched in mag-BMSC-Exos and served as a critical mediator in mag-BMSC-Exo-induced regulatory effects through inhibition of SPRY2 and activation of the PI3K/AKT and ERK1/2 signaling pathways. Conclusion: Mag-BMSC-Exos can further enhance wound healing than BMSC-Exos by improving angiogenesis and fibroblast function, and miR-21-5p upregulation in mag-BMSC-Exos might be the potential mechanism. This work offers an effective and promising protocol to improve wound healing in clinic.

SUMMARY Candida glabrata is a lethal fungal pathogen resistant to many antifungal agents and has emerged as a critical target for drug discovery. Over the past several years we have been developing a class of propargyl-linked antifolates as new antimicrobials and hypothesized that these compounds could be effective inhibitors of dihydrofolate reductase (DHFR) from C. glabrata. We initially screened a small collection of these inhibitors and found modest levels of potency. Subsequently, we determined the crystal structure of C. glabrata DHFR bound to a representative inhibitor with data to 1.6 Å resolution. Using this structure, we designed and synthesized second generation inhibitors. These new inhibitors bind the C. glabrata DHFR enzyme with subnanomolar potency, display greater than 2,000-fold levels of selectivity over the human enzyme and inhibit the growth of C. glabrata at levels observed with clinically employed antifungal therapeutics.

Follicular thyroid adenoma (FTA) precedes follicular thyroid carcinoma (FTC) by definition with a favorable prognosis compared to FTC. However, the genetic mechanism of FTA to FTC progression remains unknown. For this, it is required to disclose FTA and FTC genomes in mutational and evolutionary perspectives. We performed whole-exome sequencing and copy number profiling of 14 FTAs and 13 FTCs, which exhibited previously-known gene mutations (NRAS, HRAS, BRAF, TSHR and EIF1AX) and copy number alterations (CNAs) (22q loss and 1q gain) in follicular tumors. In addition, we found eleven potential cancer-related genes with mutations (EZH1, SPOP, NF1, TCF12, IGF2BP3, KMT2C, CNOT1, BRIP1, KDM5C, STAG2 and MAP4K3) that have not been reported in thyroid follicular tumors. Of note, FTA genomes showed comparable levels of mutations to FTC in terms of the number, sequence composition and functional consequences (potential driver mutations) of mutations. Analyses of evolutionary ages using somatic mutations as molecular clocks further identified that FTA genomes were as old as FTC genomes. Whole-transcriptome sequencing did not find any gene fusions with potential significance. Our data indicate that FTA genomes may be as old as FTC genomes, thus suggesting that follicular thyroid tumor genomes during the transition from FTA to FTC may stand stable at genomic levels in contrast to the discernable changes at pathologic and clinical levels. Also, the data suggest a possibility that the mutational profiles obtained from early biopsies may be useful for the molecular diagnosis and therapeutics of follicular tumor patients.