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Mass Spectrometry of Human Transporters.

Transporters are key to understanding how an individual will respond a particular dose of a drug. Two patients with similar systemic concentrations may have quite different local concentrations of a drug at the required site. The transporter profile of any individual depends upon a variety of genetic and environmental factors, including genotype, age, and diet status. Robust models (virtual patients) are therefore required and these models are data hungry. Necessary data include quantitative transporter profiles at the relevant organ. Liquid chromatography with tandem mass spectrometry (LC-MS/MS) is currently the most powerful method available for obtaining this information. Challenges include sourcing the tissue, isolating the hydrophobic membrane-embedded transporter proteins, preparing the samples for MS (including proteolytic digestion), choosing appropriate quantification methodology, and optimizing the LC-MS/MS conditions. Great progress has been made with all of these, especially within the last few years, and is discussed here. Expected final online publication date for the , Volume 13 is June 12, 2020. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

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Inner-Stress-Optimized High-Density Fe3O4 Dots Embedded in Graphitic Carbon Layers with Enhanced Lithium Storage.

The volume variation of electrode materials will lead to poor cyclability of lithium-ion batteries (LIBs) during lithiation/delithiation process. Instead, inner stress fragmentation is creatively used to change carbon-layer-capped Fe3O4 particles ~30 nm in diameter into high-density Fe3O4 dots ~4 nm in size embedded in ultrathin carbon layers. The optimized structure shows a remarkable 45.2 % enhancement of lithium storage from 804.7 (the 10th cycle) to 1168.7 mA h g-1 (the 250th cycle) at 500 mA g-1, even retaining 1239.5 mA h g-1 after another 550 cycles. The electrochemical measurements reveal the enhanced capacitive behavior of the high-density Fe3O4 [email protected] layers, which has more extra active sites for the insertion/extraction of Li+ ions confirmed by the differential capacity plots (DCPs), leading to remarkably increased specific capacity during cycling. The restructured electrode also shows a superior rate capacity and excellent cycling stability (938.7 and 815.4 mA h g-1 over 2000 cycles at 1000 and 2000 mA g-1, respectively). XPS and TEM characterizations show the optimized structure has stable structural and componental stability even at large rates. This work presents a MOF-guided synthesis of high-density Fe3O4-dots anode material optimized by inner stress fragmentation, showing a feasible route to design high-efficient electrode materials.

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High-loading boron nitride-based bioinspired paper with plastic-like ductility and metal-like thermal conductivity.

Although desirable in next-generation flexible electronics, fabricating hybrid film materials with excellent integration of mechanical, thermally conductive yet electrically insulating properties is still under challenge. In mollusk nacre, a small volume of chitin nanofiber framework hosts 95 vol% CaCO3 microplatelets, enabling the high-loading natural composites to exhibit ductile deformation behavior. Inspired by this, we fabricate a large-area, boron nitride-based bioinspired paper by a facile sol-gel-film conversion approach, in which BN microplatelets with a loading of 40-80 wt% are embedded into 3D poly(p-phenylene benzobisoxazole) nanofiber framework. Due to the vital role of 3D nanofiber framework, the BN-based paper exhibits plastic-like ductility (38-80%), ultrahigh toughness (10-100 MJ m-3) and good folding endurance. The high-loading BN platelets form an oriented, percolative network, and impart the paper with outstanding in-plane thermal conductivity (77.1-214.2 W m-1 K-1), comparable to some metals, such as aluminum alloys (108-230 W m-1 K-1). Using the electrically insulating BN-based paper as a flexible substrate, we demonstrate its promising application for lowering the temperature of electronic device.

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Automated platform for cell selection and separation based on four-dimensional motility and matrix degradation.

Motility and invasion are key steps in the metastatic cascade, enabling cells to move through normal tissue borders into the surrounding stroma. Most available in vitro assays track cell motility or cell invasion but lack the ability to measure both simultaneously and then separate single cells with unique behaviors. In this work, we developed a cell-separation platform capable of tracking cell movement (chemokinesis) and invasion through an extracellular matrix in space and time. The platform utilized a collagen scaffold with embedded tumor cells overlaid onto a microraft array. Confocal microscopy enabled high resolution (0.4 × 0.4 × 3.5 µm voxel) monitoring of cell movement within the scaffolds. Two pancreatic cancer cell lines with known differing invasiveness were characterized on this platform, with median motilities of 14 ± 6 μm and 10 ± 4 μm over 48 h. Within the same cell line, cells demonstrated highly variable motility, with XYZ movement ranging from 144 μm to 2 μm over 24 h. The ten lowest and highest motility cells, with median movements of 33 ± 11 μm and 3 ± 1 μm, respectively, were separated and sub-cultured. After 6 weeks of culture, the cell populations were assayed on a Transwell invasion assay and 227 ± 56 cells were invasive in the high motility population while only 48 ± 10 cells were invasive in the low motility population, indicating that the resulting offspring possessed a motility phenotype reflective of the parental cells. This work demonstrates the feasibility of sorting single cells based on complex phenotypes along with the capability to further probe those cells and explore biological phenomena.

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Characterizations of Gene Alterations in Melanoma Patients from Chinese Population.

Melanoma is a human skin malignant tumor with high invasion and poor prognosis. The limited understanding of genomic alterations in melanomas in China impedes the diagnosis and therapeutic strategy selection. We conducted comprehensive genomic profiling of melanomas from 39 primary and metastatic formalin-fixed paraffin-embedded (FFPE) samples from 27 patients in China based on an NGS panel of 223 genes. No significant difference in gene alterations was found between primary and metastasis melanomas. The status of germline mutation, CNV, and somatic mutation in our cohort was quite different from that reported in Western populations. We further delineated the mutation patterns of 4 molecular subgroups (BRAF, RAS, NF1, and Triple-WT) of melanoma in our cohort. BRAF mutations were more frequently identified in melanomas without chromic sun-induced damage (non-CSD), while RAS mutations were more likely observed in acral melanomas. NF1 and Triple-WT subgroups were unbiased between melanomas arising in non-CSD and acral skin. BRAF, RAS, and NF1 mutations were significantly associated with lymph node metastasis or presence of ulceration, implying that these cancer driver genes were independent prognostic factors. In summary, our results suggest that mutational profiles of malignant melanomas in China are significantly different from Western countries, and both gene mutation and amplification play an important role in the development and progression of melanomas.

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Polarization of Macrophages in Epidural Inflammation Induced by Canine Intervertebral Disc Herniation.

Canine interverterbral disc (IVD), although physiologically acellular, displays an inflammatory cell population consisting almost exclusively of macrophages (Mϕ) when acutely herniated. Mϕ encompass a heterogenous cell population, roughly divided into classically (M1) or alternatively activated (M2)Mϕ. Polarization into M1 Mϕ leads to strong antimicrobial activity and pro-inflammatory response. In contrast, M2Mϕ exibit anti-inflammatory function and regulate wound healing. The purpose of this study was to characterize the phenotype of the Mϕ population present in naturally occurring IVD herniation. IVD material of dogs with IVD disease was collected during standard decompressive surgery. A negative control consisting of IVD material of dogs without IVD degeneration and a positive control consisting of canine liver and lymph node samples were also included. All samples were embedded in OCT and shock frozen. Eight micrometer cryostat sections were prepared, air dried and immunostained without prefixation or permeabilization. CD14 was used as marker Mϕ, MHCII for M1Mϕ and CD206 for M2Mϕ. Fifteen samples of dogs with IVD herniation, 10 negative, and 5 positive control samples were obtained. No positive cell was found in the negative control group. The positive control group displayed several MHCII and CD206 positive cells, all of them being simultaneously positive to CD14. All herniated samples displayed a mixed population of M1Mϕ and M2Mϕ, and some sparse Mϕ displaying markers for both M1 and M2Mϕ simultaneously. The mixed phenotype encountered shows the plasticity and dynamism of Mϕ and evidences the chronic component of IVD disease despite its acute clinical presentation.

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Preclinical activity of sacituzumab govitecan (IMMU-132) in uterine and ovarian carcinosarcomas.

Uterine and ovarian carcinosarcomas (CS) are rare cancers with poor prognosis. Sacituzumab-govitecan (SG) is a new class of antibody-drug-conjugate (ADC) targeting the human-trophoblast-cell-surface marker (Trop-2) conjugated with the active metabolite of irinotecan (SN-38). We evaluated the efficacy of SG against biologically aggressive CS.

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Optimization of ATP Synthase c-Rings for Oxygenic Photosynthesis.

The conversion of sunlight into useable cellular energy occurs the proton-coupled electron transfer reactions of photosynthesis. Light is absorbed by photosynthetic pigments and transferred to photochemical reaction centers to initiate electron and proton transfer reactions to store energy in a redox gradient and an electrochemical proton gradient (proton motive force, ), composed of a concentration gradient (ΔpH) and an electric field (Δ), which drives the synthesis of ATP through the thylakoid FF-ATP synthase. Although ATP synthase structure and function are conserved across biological kingdoms, the number of membrane-embedded ion-binding subunits varies between organisms, ranging from 8 to 17, theoretically altering the H/ATP ratio for different ATP synthase complexes, with profound implications for the bioenergetic processes of cellular metabolism. Of the known -ring stoichiometries, photosynthetic -rings are among the largest identified stoichiometries, and it has been proposed that decreasing the c-stoichiometry could increase the energy conversion efficiency of photosynthesis. Indeed, there is strong evidence that the high H/ATP of the chloroplast ATP synthase results in a low ATP/nicotinamide adenine dinucleotide phosphate (NADPH) ratio produced by photosynthetic linear electron flow, requiring secondary processes such as cyclic electron flow to support downstream metabolism. We hypothesize that the larger subunit stoichiometry observed in photosynthetic ATP synthases was selected for because it allows the thylakoid to maintain in a range where ATP synthesis is supported, but avoids excess Δ and ΔpH, both of which can lead to production of reactive oxygen species and subsequent photodamage. Numerical kinetic simulations of the energetics of chloroplast photosynthetic reactions with altered -ring size predicts the energy storage of and its effects on the photochemical reaction centers strongly support this hypothesis, suggesting that, despite the low efficiency and suboptimal ATP/NADPH ratio, a high H/ATP is favored to avoid photodamage. This has important implications for the evolution and regulation of photosynthesis as well as for synthetic biology efforts to alter photosynthetic efficiency by engineering the ATP synthase.

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Nanofabrication for all-soft and high-density electronic devices based on liquid metal.

Innovations in soft material synthesis and fabrication technologies have led to the development of integrated soft electronic devices. Such soft devices offer opportunities to interact with biological cells, mimicking their soft environment. However, existing fabrication technologies cannot create the submicron-scale, soft transducers needed for healthcare and medical applications involving single cells. This work presents a nanofabrication strategy to create submicron-scale, all-soft electronic devices based on eutectic gallium-indium alloy (EGaIn) using a hybrid method utilizing electron-beam lithography and soft lithography. The hybrid lithography process is applied to a biphasic structure, comprising a metallic adhesion layer coated with EGaIn, to create soft nano/microstructures embedded in elastomeric materials. Submicron-scale EGaIn thin-film patterning with feature sizes as small as 180 nm and 1 μm line spacing was achieved, resulting in the highest resolution EGaIn patterning technique to date. The resulting soft and stretchable EGaIn patterns offer a currently unrivaled combination of resolution, electrical conductivity, and electronic/wiring density.

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