Search

SUMMARYOne of the earliest pathophysiological perturbations in Alzheimer’s Disease (AD) may arise from dysfunction of fast-spiking parvalbumin (PV) interneurons (PV-INs). Defining early protein-level (proteomic) alterations in PV-INs can provide key biological and translationally relevant insights. Here, we use cell-type-specific in vivo biotinylation of proteins (CIBOP) coupled with mass spectrometry to obtain native-state proteomes of PV interneurons. PV-INs exhibited proteomic signatures of high metabolic, mitochondrial, and translational activity, with over-representation of causally linked AD genetic risk factors. Analyses of bulk brain proteomes indicated strong correlations between PV-IN proteins with cognitive decline in humans, and with progressive neuropathology in humans and mouse models of Aβ pathology. Furthermore, PV-IN-specific proteomes revealed unique signatures of increased mitochondrial and metabolic proteins, but decreased synaptic and mTOR signaling proteins in response to early Aβ pathology. PV-specific changes were not apparent in whole-brain proteomes. These findings showcase the first native state PV-IN proteomes in mammalian brain, revealing a molecular basis for their unique vulnerabilities in AD.

The importance of mitogen-activated protein kinase (MAPK) pathway signaling in regulating microglia-mediated neuroinflammation in Alzheimer’s disease (AD) remains unclear. We examined the role of MAPK signaling in microglia using a preclinical model of AD pathology and quantitative proteomics studies of postmortem human brains. In multiplex immunoassay analyses of MAPK phosphoproteins in acutely isolated microglia and brain tissue from 5xFAD mice, we found phosphorylated extracellular signal-regulated kinase (ERK) was the most strongly upregulated phosphoprotein within the MAPK pathway in acutely isolated microglia, but not whole-brain tissue from 5xFAD mice. The importance of ERK signaling in primary microglia cultures was next investigated using transcriptomic profiling and functional assays of amyloid-β and neuronal phagocytosis, which confirmed that ERK is a critical regulator of IFNγ-mediated pro-inflammatory activation of microglia, although it was also partly important for constitutive microglial functions. Phospho-ERK was an upstream regulator of disease-associated microglial gene expression (Trem2, Tyrobp), as well as several human AD risk genes (Bin1, Cd33, Trem2, Cnn2), indicative of the importance of microglial ERK signaling in AD pathology. Quantitative proteomic analyses of post-mortem human brain showed that ERK1 and ERK2 were the only MAPK proteins with increased protein expression and positive associations with neuropathological grade. In a human brain phosphoproteomic study, we found evidence for increased flux through the ERK signaling pathway in AD. Overall, our analyses strongly suggest that ERK phosphorylation, particularly in microglia in mouse models, is a regulator of pro-inflammatory immune responses in AD pathogenesis.

This study adopts the nonlinear time-varying factor model and the Mann–Kendall non-parametric rank-sequence test to assess the convergence of non-financial listed companies’ debt leverage in China ...

MnSiO3 nanoparticles (NPs) were prepared by precipitation method, their structure and composition were characterised by transmission electron microscopy, electron diffraction spectroscopy, X-ray diffraction, and X-ray photoelectron spectroscopy. Due to the oxidase-like enzyme activity of MnSiO3, it can catalyse the substrate 3,3′,5,5′-tetramethylbenzidine (TMB) in acidic environment to form an oxidised TMB which has an absorption peak at 652 nm in the absence of hydrogen peroxide. When Fe2+ was added to the assay, the oxidase-like activity of MnSiO3 was inhibited, and the intensity of the absorption at 652 nm was reduced. Based on the high oxidase-like activity, a colorimetric method for Fe2+ detection was established, it showed high selectivity and sensitivity. In the optimised conditions, the limit of detection was 0.5 μM with a linear range from 3 to 63 μM. Moreover, the method was applied to determine Fe2+ in spiked water samples which gave satisfactory recoveries.

Isolation and proteomic profiling of brain cell types, particularly neurons, pose several technical challenges which limit our ability to resolve distinct cellular phenotypes in neurological diseases. Therefore, we generated a novel mouse line that enables cell type-specific expression of a biotin ligase, TurboID, via Cre-lox strategy for in vivo proximity-dependent biotinylation of proteins. Using adenoviral-based and transgenic approaches, we show striking protein biotinylation in neuronal cell bodies and axons throughout the mouse brain. We quantified more than 2,000 neuron-derived proteins following enrichment that mapped to numerous subcellular compartments. Synaptic, transmembrane transporters, ion channel subunits, and disease-relevant druggable targets were among the most significantly enriched proteins. Remarkably, we resolved brain region-specific proteomic profiles of Camk2a neurons with distinct functional molecular signatures and disease associations that may underlie regional neuronal vulnerability. Leveraging the neuronal specificity of this in vivo biotinylation strategy, we used an antibody-based approach to uncover regionally unique patterns of neuron-derived signaling phospho-proteins and cytokines, particularly in the cortex and cerebellum. Our work provides a proteomic framework to investigate cell type-specific mechanisms driving physiological and pathological states of the brain as well as complex tissues beyond the brain.

Proteomic profiling of brain cell types using isolation-based strategies pose limitations in resolving cellular phenotypes representative of their native state. We describe a mouse line for cell type-specific expression of biotin ligase TurboID, for in vivo biotinylation of proteins. Using adenoviral and transgenic approaches to label neurons, we show robust protein biotinylation in neuronal soma and axons throughout the brain, allowing quantitation of over 2000 neuron-derived proteins spanning synaptic proteins, transporters, ion channels and disease-relevant druggable targets. Next, we contrast Camk2a-neuron and Aldh1l1-astrocyte proteomes and identify brain region-specific proteomic differences within both cell types, some of which might potentially underlie the selective vulnerability to neurological diseases. Leveraging the cellular specificity of proteomic labeling, we apply an antibody-based approach to uncover differences in neuron and astrocyte-derived signaling phospho-proteins and cytokines. This approach will facilitate the characterization of cell-type specific proteomes in a diverse number of tissues under both physiological and pathological states.

Kv1.3 potassium channels, expressed by proinflammatory central nervous system mononuclear phagocytes (CNS-MPs), are promising therapeutic targets for modulating neuroinflammation in Alzheimer’s disease (AD). The molecular characteristics of Kv1.3-high CNS-MPs and their cellular origin from microglia or CNS-infiltrating monocytes are unclear. While Kv1.3 blockade reduces amyloid beta (Aβ) burden in mouse models, the downstream immune effects on molecular profiles of CNS-MPs remain unknown. We show that functional Kv1.3 channels are selectively expressed by a subset of CD11b(+)CD45(+) CNS-MPs acutely isolated from an Aβ mouse model (5xFAD) as well as fresh postmortem human AD brain. Transcriptomic profiling of purified CD11b(+)Kv1.3(+) CNS-MPs, CD11b(+)CD45(int) Kv1.3(neg) microglia, and peripheral monocytes from 5xFAD mice revealed that Kv1.3-high CNS-MPs highly express canonical microglial markers (Tmem119, P2ry12) and are distinct from peripheral Ly6c(high)/Ly6c(low) monocytes. Unlike homeostatic microglia, Kv1.3-high CNS-MPs express relatively lower levels of homeostatic genes, higher levels of CD11c, and increased levels of glutamatergic transcripts, potentially representing phagocytic uptake of neuronal elements. Using irradiation bone marrow CD45.1/CD45.2 chimerism in 5xFAD mice, we show that Kv1.3(+) CNS-MPs originate from microglia and not blood-derived monocytes. We show that Kv1.3 channels regulate membrane potential and early signaling events in microglia. Finally, in vivo blockade of Kv1.3 channels in 5xFAD mice by ShK-223 reduced Aβ burden, increased CD11c(+) CNS-MPs, and expression of phagocytic genes while suppressing proinflammatory genes (IL1b). Our results confirm the microglial origin and identify unique molecular features of Kv1.3-expressing CNS-MPs. In addition, we provide evidence for CNS immunomodulation by Kv1.3 blockers in AD mouse models resulting in a prophagocytic phenotype.

Au@AgPt yolk-shell nanoparticles (Y-SNPs) with Au core and porous Ag/Pt alloy shell were prepared via a double-template method. This synthesis employed Au@Ag core-shell nanoparticles as a hard-template to obtain Au@AgPt Y-SNPs by a galvanic replacement reaction between PtCl62− and Ag. Brij-58 surfactant micelles served as a soft-template to prepare dendritic and porous shell. The thus-synthesized Au@AgPt Y-SNPs exhibited enhanced catalytic performance (889 mA mgPt−1) and stability for methanol oxidation reaction (MOR) at room temperature, which is 5 times as high as the commercial Pt/C (20 wt% Pt). The enhanced electrocatalytic activity is attributed to following aspects: electronic effect between Pt and Au, synergism between Pt and Ag, and nanoreactor confinement effect in the cavity between the Au core and the shell. The present work provides a simple strategy for the formation of ternary metallic Y-SNPs, which exhibit promising applications in electrocatalysis, energy conversion and sensing.

Background Proteomic characterization of microglia provides the most proximate assessment of functionally relevant molecular mechanisms of neuroinflammation. However, microglial proteomics studies have been limited by low cellular yield and contamination by non-microglial proteins using existing enrichment strategies. Methods We coupled magnetic-activated cell sorting (MACS) and fluorescence activated cell sorting (FACS) of microglia with tandem mass tag-mass spectrometry (TMT-MS) to obtain a highly-pure microglial proteome and identified a core set of highly-abundant microglial proteins in adult mouse brain. We interrogated existing human proteomic data for Alzheimer’s disease (AD) relevance of highly-abundant microglial proteins and performed immuno-histochemical and in-vitro validation studies. Results Quantitative multiplexed proteomics by TMT-MS of CD11b + MACS-enriched (N = 5 mice) and FACS-isolated (N = 5 mice), from adult wild-type mice, identified 1791 proteins. A total of 203 proteins were highly abundant in both datasets, representing a core-set of highly abundant microglial proteins. In addition, we found 953 differentially enriched proteins comparing MACS and FACS-based approaches, indicating significant differences between both strategies. The FACS-isolated microglia proteome was enriched with cytosolic, endoplasmic reticulum, and ribosomal proteins involved in protein metabolism and immune system functions, as well as an abundance of canonical microglial proteins. Conversely, the MACS-enriched microglia proteome was enriched with mitochondrial and synaptic proteins and higher abundance of neuronal, oligodendrocytic and astrocytic proteins. From the 203 consensus microglial proteins with high abundance in both datasets, we confirmed microglial expression of moesin (Msn) in wild-type and 5xFAD mouse brains as well as in human AD brains. Msn expression is nearly exclusively found in microglia that surround Aβ plaques in 5xFAD brains. In in-vitro primary microglial studies, Msn silencing by siRNA decreased Aβ phagocytosis and increased lipopolysaccharide-induced production of the pro-inflammatory cytokine, tumor necrosis factor (TNF). In network analysis of human brain proteomic data, Msn was a hub protein of an inflammatory co-expression module positively associated with AD neuropathological features and cognitive dysfunction. Conclusions Using FACS coupled with TMT-MS as the method of choice for microglial proteomics, we define a core set of highly-abundant adult microglial proteins. Among these, we validate Msn as highly-abundant in plaque-associated microglia with relevance to human AD.

BackgroundThe mitogen-activated protein kinase (MAPK) pathway is a central regulator of gene expression, pro-survival signaling, and inflammation. However, the importance of MAPK pathway signaling in regulating microglia-mediated neuroinflammation in Alzheimer’s Disease (AD) remains unclear. Here we examined the role of MAPK signaling in microglia using pre-clinical in-vitro and in-vivo models of AD pathology integrated with quantitative proteomics studies of post-mortem human brains.MethodsWe performed multiplexed immunoassay analyses of MAPK phosphoproteins, particularly ERK1/2, in acutely-isolated microglia and brain tissue from wild-type and 5xFAD mice. Neuropathological studies of mouse and human brain tissues were performed to quantify total and phosphorylated ERK protein in AD. The importance of ERK signaling in unstimulated and interferon γ (IFNγ)-stimulated primary microglia cultures was investigated using NanoString transcriptomic profiling, coupled with functional assays of amyloid β (Aβ) and neuronal phagocytosis. Receptor tyrosine kinases (RTKs) likely responsible for ERK signaling in homeostatic microglia and disease-associated-microglia (DAM) states and ERK-regulated human AD risk genes were identified using gene expression data. Total and phosphorylated MAPKs in human post-mortem brain tissues were measured in quantitative proteomic datasets.ResultsPhosphorylated ERK was the most strongly up-regulated signaling protein within the MAPK pathway in microglia acutely isolated from 5xFAD brains. Neuroinflammatory transcriptomic and phagocytic profiling of mouse microglia confirmed that ERK is a critical regulator of IFNγ-mediated pro-inflammatory activation of microglia, although it was also important for constitutive microglial functions. Phospho-ERK was an upstream regulator of disease-associated microglia (DAM) gene expression (Trem2, Tyrobp), as well as of several human AD risk genes (Bin1, Cd33, Trem2, Cnn2). Among RTKs that signal via ERK, CSF1R and MERTK were primarily expressed by homeostatic microglia while AXL and FLT1 were likely regulators of ERK signaling in DAM. Within DAM, FLT4 and IGF1R were specifically expressed by pro- and anti-inflammatory DAM sub-profiles respectively. In quantitative proteomic analyses of post-mortem human brains from non-disease, asymptomatic and cognitively-impaired AD cases, ERK1 and ERK2 were the only MAPK pathway signaling proteins with increased protein expression and positive associations with neuropathological grade. Moreover, in a phospho-proteomic study of post-mortem human brains from controls, asymptomatic and symptomatic AD cases, we found evidence for a progressive increased flux through the ERK signaling pathway.ConclusionsOur integrated analyses using pre-clinical models and human proteomic data strongly suggest that ERK phosphorylation in microglia is a critical regulator of pro-inflammatory immune response in AD pathogenesis and that modulation of ERK via upstream RTKs may reveal novel avenues for immunomodulation.

Plasmonic metal nanocatalysts have excellent light trapping properties and high chemical reactivity. Impressively, Au nanostructures can absorb a wide array of visible light by tuning their morphology. In this work, spherical gold nanoparticles (Au NSs), multi-branched gold nanoparticles (Au NMs) and gold nanorods (Au NRs) were successfully synthesized; the shape- and size-dependences of these gold nanocatalysts on the methanol oxidation reaction (MOR) under light irradiation were studied. It is worth mentioning that Au NRs have the highest anode peak current density under dark conditions due to the exposure of highly active facets. A similar enhancement effect was obtained for Au NSs and Au NMs under visible light irradiation, which is due to the generation of a high concentration of energetic charge carriers on these Au nanostructures. The size dependences of Au NSs on the MOR showed that a larger electrochemically active surface area (ECSA) was obtained for small nanoparticles, which is due to the surface effect. In addition, the catalytic performance, durability and anti-CO stripping of these Au nanocatalysts under visible light irradiation, as well as the effect of light intensity and wavelength were described in detail. This work provides an insight into the mechanism of plasmon enhanced electrocatalysis by Au nanostructures with different sizes and shapes.

Background Microglia are innate immune cells of the brain that perform phagocytic and inflammatory functions in disease conditions. Transcriptomic studies of acutely-isolated microglia have provided novel insights into their molecular and functional diversity in homeostatic and neurodegenerative disease states. State-of-the-art mass spectrometry methods can comprehensively characterize proteomic alterations in microglia in neurodegenerative disorders, potentially providing novel functionally relevant molecular insights that are not provided by transcriptomics. However, comprehensive proteomic profiling of adult primary microglia in neurodegenerative disease conditions has not been performed. Methods We performed quantitative mass spectrometry based proteomic analyses of purified CD11b+ acutely-isolated microglia from adult (6 mo) mice in normal, acute neuroinflammatory (LPS-treatment) and chronic neurodegenerative states (5xFAD model of Alzheimer’s disease [AD]). Differential expression analyses were performed to characterize specific microglial proteomic changes in 5xFAD mice and identify overlap with LPS-induced pro-inflammatory changes. Our results were also contrasted with existing proteomic data from wild-type mouse microglia and from existing microglial transcriptomic data from wild-type and 5xFAD mice. Neuropathological validation studies of select proteins were performed in human AD and 5xFAD brains. Results Of 4133 proteins identified, 187 microglial proteins were differentially expressed in the 5xFAD mouse model of AD pathology, including proteins with previously known (Apoe, Clu and Htra1) as well as previously unreported relevance to AD biology (Cotl1 and Hexb). Proteins upregulated in 5xFAD microglia shared significant overlap with pro-inflammatory changes observed in LPS-treated mice. Several proteins increased in human AD brain were also upregulated by 5xFAD microglia (Aβ peptide, Apoe, Htra1, Cotl1 and Clu). Cotl1 was identified as a novel microglia-specific marker with increased expression and strong association with AD neuropathology. Apoe protein was also detected within plaque-associated microglia in which Apoe and Aβ were highly co-localized, suggesting a role for Apoe in phagocytic clearance of Aβ. Conclusions We report a comprehensive proteomic study of adult mouse microglia derived from acute neuroinflammation and AD models, representing a valuable resource to the neuroscience research community. We highlight shared and unique microglial proteomic changes in acute neuroinflammation aging and AD mouse models and identify novel roles for microglial proteins in human neurodegeneration. Electronic supplementary material The online version of this article (10.1186/s13024-018-0266-4) contains supplementary material, which is available to authorized users.

Multifunctional enzymatic nanomaterials (nanozymes) with good biocompatibility, strong enzymatic activity, and substrate specificity are the desired alternatives to natural enzymes. Here, the design of a peroxidase mimetic nanozyme capable of detecting and depleting glutathione (GSH) in the bacterial cells is presented. The nanozyme palladium-iron nanostructure decorated graphdiyne nanosheet (PdFe/GDY) was prepared by a facile hydrothermal approach. It can efficiently catalyze the decomposition of H2O2 to produce •OH by a peroxidase-like mechanism. Surprisingly, the PdFe/GDY nanozymes can detect GSH in bacteria with excellent specificity and ultra-low detection limit of 24.45 nM. Moreover, the PdFe/GDY nanozymes can consume GSH for bacterial disinfection with killing efficacy > 99.99%, promote wound healing and do not cause significant toxicity in vitro and in vivo. To our knowledge, this is the first example of a GDY-based nanozyme biosensor and antibacterial agent. Collectively, this work would be indicative to construct GDY-based nanozymes in clinical diagnosis and sterilization.

In this work, the PtCu electrocatalyst supported on graphdiyne (PtCu/GDY) nanocomposite was synthesized for methanol oxidation reaction (MOR). The results demonstrated that the obtained PtCu/GDY catalyst has a good dispersion on graphdiyne. The electrochemical experiments indicated that the PtCu/GDY displayed a superior electrocatalytic activity towards MOR with a high mass activity of 336 mA mg−1. Furthermore, the presence of GDY can significantly improve the CO tolerance property of PtCu for MOR, which facilitating OHads generation from H2O decomposition. Therefore, the PtCu/GDY electrocatalyst offers an exciting opportunity to be commercialization of direct methanol fuel cells.

Hollow dendritic Ag/Pt alloy nanoparticles were synthesized by a double template method: Ag nanoparticles as the hard template to obtain hollow spheres by a galvanic replacement reaction between PtCl62− and metallic Ag and surfactant micelles (Brij58) as the soft template to generate porous dendrites. The formation of a Ag/Pt alloy phase was confirmed by XRD and HRTEM. Elemental mapping and line scanning revealed the formation of the hollow architecture. We studied the effects of the Ag/Pt ratio, surfactant and reaction temperature on the morphology. In addition, we explored the formation process of hollow dendritic Ag/Pt nanoparticles by tracking the morphologies of the nanostructures formed at different stages. In order to improve the electrocatalytic property, we controlled the size of the nanoparticles and the thickness of the shell by adjusting the amount of the precursor. We found that these Ag/Pt alloy nanoparticles exhibited high activity (440 mA mg−1) and stability as an electrocatalyst for catalyzing methanol oxidation.

A Schiff base derivative, N-(3-methoxy-4-hydroxybenzylidene)salicyloylhydrazone (L), shows high selectivity to Lu3+ not only in abiotic systems but also in living cells with fluorescence. The presence of PVP can increase the fluorescence intensity. The fluorescence enhancement is presumably due to a combinational effect of intramolecular charge transfer (ICT) and excited-state intra/intermolecular proton transfer (ESPT) during the chelation of L to Lu3+ in a 2 : 1 complex mode.

Microglia transform from homeostatic to disease-associated-microglia (DAM) profiles in neurodegeneration. Within DAM, we recently identified distinct pro-inflammatory and anti-inflammatory sub-profiles although the transcriptional regulators of homeostatic and distinct DAM gene profiles in microglia remain unclear. Informed by our previous studies, we nominated CEBPα, IRF1 and LXRβ as likely regulators of homeostatic, pro-inflammatory and anti-inflammatory DAM states and performed in-vitro siRNA studies in primary microglia to identify the role of each TF in regulating microglial activation, using an integrated transcriptomics, bioinformatics and experimental validation approach. Efficient (>70%) silencing of each TF in primary microglia revealed reciprocal regulation between each TF specifically following pro-inflammatory activation. Neuroinflammatory transcriptomic profiling of microglia coupled with qPCR validation revealed distinct gene clusters with unique patterns of regulation by each TF which were independent of LPS stimulation. While all three TFs (especially IRF1 and LXRβ) positively regulated core DAM genes (Apoe, Axl, Clec7a, Tyrobp and Trem2) as well as homeostatic and pro-inflammatory DAM genes, LPS and IFNγ increased pro-inflammatory DAM but suppressed homeostatic and anti-inflammatory DAM gene expression via an Erk1/2-dependent signaling pathway. IRF1 and LXRβ silencing suppressed microglial phagocytic activity for macroparticles (polystyrene microspheres) as well as fAβ42 while IRF1 silencing strongly suppressed production of pro-inflammatory cytokines in response to LPS. Our studies reveal complex transcriptional regulation of homeostatic microglia and DAM profiles whereby IRF1, LXRβ and CEBPα positively regulate both pro- and anti-inflammatory DAM genes while activating stimuli independently augment pro-inflammatory DAM responses and suppress homeostatic and anti-inflammatory responses via Erk signaling. This framework can guide development of therapeutic immuno-modulatory strategies for neurodegeneration.

Background Microglia and CNS-infiltrating monocytes/macrophages (CNS-MPs) perform pro-inflammatory and protective anti-inflammatory functions following ischemic stroke. Selective inhibition of pro-inflammatory responses can be achieved by Kv1.3 channel blockade, resulting in a lower infarct size in the transient middle cerebral artery occlusion (tMCAO) model. Whether beneficial effects of Kv1.3 blockers are mediated by targeting microglia or CNS-infiltrating monocytes/macrophages remains unclear. Methods In the 30-min tMCAO mouse model, we profiled functional cell-surface Kv1.3 channels and phagocytic properties of acutely isolated CNS-MPs at various timepoints post-reperfusion. Kv1.3 channels were flow cytometrically detected using fluorescein-conjugated Kv1.3-binding peptide ShK-F6CA as well as by immunohistochemistry. Quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) was performed to measure Kv1.3 (Kcna3) and Kir2.1 (Kcnj2) gene expression. Phagocytosis of 1-μm microspheres by acutely isolated CNS-MPs was measured by flow cytometry. Results In flow cytometric assays, Kv1.3 channel expression by CD11b+ CNS-MPs was increased between 24 and 72 h post-tMCAO and decreased by 7 days post-tMCAO. Increased Kv1.3 expression was restricted to CD11b+CD45lowLy6clow (microglia) and CD11b+CD45highLy6Clow CNS-MPs but not CD11b+CD45highLy6chigh inflammatory monocytes/macrophages. In immunohistochemical studies, Kv1.3 protein expression was increased in Iba1+ microglia at 24-48 h post-tMCAO. No change in Kv1.3 mRNA in CNS-MPs was observed following tMCAO. Conclusions We conclude that resident microglia and a subset of CD45highLy6clow CNS-MPs are the likely cellular targets of Kv1.3 blockers and the delayed phase of neuroinflammation is the optimal therapeutic window for Kv1.3 blockade in ischemic stroke. Electronic supplementary material The online version of this article (10.1186/s12974-019-1510-8) contains supplementary material, which is available to authorized users.

Etched PtCu nanowires (NWs) were synthesized by a hydrothermal reaction and chemical etching process. The NWs are shown to be viable peroxidase (POx) mimics capable of catalyzing the oxidation of the substrate 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of H2O2 to form a blue-green coloration. The mechanism of catalysis was investigated and the results demonstrated that H2O2 is decomposed to form hydroxyl radicals which oxidize TMB in the presence of the NWs. Under optimized conditions, a steady-state kinetic analysis revealed that the NWs possess a stronger affinity for H2O2 and TMB compared to the enzyme horseradish POx. Based on the high POx-like activity, a colorimetric assay for H2O2 was established. Absorbance at 652 nm increases linearly in the 0.1–300 μM H2O2 concentration range, and the detection limit is 0.06 μM (at S/N = 3). The assay was successfully applied to the determination of H2O2 in (spiked) milk and contact lens solution. Furthermore, a highly sensitive test strip was designed which represents a low cost and fast alternative for the visual determination of H2O2.

Background: Neuroinflammation mediated by microglia and CNS-infiltrating macrophages (CNS-MPs) impacts neuronal survival and recovery following ischemic stroke. Transcriptomic profiling of microglia has revealed switching from homeostatic to disease-associated microglial (DAM) states in neurodegeneration but microglial profiles at acute/subacute phases post-stroke remain unclear. Methods: We performed transcriptomics (770 gene panel) of acutely-isolated CNS-MPs from ipsi- and contralateral hemispheres following transient (30min) middle cerebral artery occlusion (tMCAO) in adult C57BL6J mice at 24-72h post-tMCAO. Normalized data were used for differential expression analyses to identify changes distinct to the ipsilateral hemisphere as well as to delayed stages post-MCAO (>±2-fold change, p Results: Among 533 genes included, highly abundant transcripts in our dataset were enriched (p=0.003) for abundantly expressed microglial genes, confirming microglial enrichment. 52 transcripts were differentially expressed ipsilaterally (22 upregulated, 29 downregulated) of which 17 changed 2-fold (7 upregulated: Spp1, Ccl2, Lilrb4a, Anapc15; 10 downregulated: Ppfia4, Trpm4, Reln, Ldlrad3, Camk4). Ipsilaterally, delayed differential expression was observed for 37 genes (33 upregulated including Apoe Fabp5, Il1b and Grn; 4 downregulated) suggesting an emerging DAM post-tMCAO, further corroborated by quantitative RT-PCR for DAM genes (Itgax, Timp2, Il1b, Ptgs2). Conclusions: Early and delayed CNS-MP transcriptomic profiles are significantly different with evidence for emergence of DAM after 72h post-ischemia. We identify microglial osteopontin (Spp1) as being highly upregulated post-MCAO and support roles for neurodegeneration-associated genes (Apoe, Grn) in delayed post-stroke neuroinflammation.

Figure S3. Minimal cell-surface Kv1.3 expression by splenic monocytes. (DOCX 475 kb)

A facile method to prepare Pt-Cu nanowires (NWs) was introduced. Structural characterization such as high-resolution transmission electron microscope (HR-TEM), selected-area electron diffraction (SAED), EDS element mapping, X-ray photoelectron spectroscopy (XPS), X-ray diffraction (XRD), and inductively coupled plasma mass spectrometry (ICP-MS) showed the formation of Pt-Cu alloy, with a width of 4.5 nm on average. The formation process of Pt-Cu NWs was studied; it was found that bromine ion, who has preferential adsorption on Pt (100) face, served as a growth-directing agent; Brij58 not only served as a protector but also played an important role in forming Pt-Cu NWs; the mechanism was proposed. Their electrocatalytic activity towards methanol oxidation was investigated; we found that the current density of Pt-Cu NWs was 295 mA·mg-1 when the ratio of Pt/Cu is 1 : 1, which is 11.5 and 2.35 times higher than that of pure Pt (26 mA·mg-1) and commercial Pt/C (126 mA·mg-1). The high electrocatalytic activity is attributed to the presence of abundant structural defects and surface active sites on the synthesized Pt-Cu NWs.

Figure S5. Correlation between functional Kv1.3 channel expression and phagocytic capacity for fluorescent microspheres in acutely isolated CNS-MPs following tMCAO. (DOCX 26 kb)

Figure S2. Alterations in proportions of CD45high CNS-MP subsets following tMCAO. (DOCX 319 kb)

Figure S4. Immunohistochemical validation of Kv1.3 protein expression by microglia following tMCAO (related to Fig. 3). (DOCX 3630 kb)

Dendritic Pt–Cu nanoparticles were synthesized by a facile one-step method with the help of surfactant Brij58 at room temperature, and we also studied the effects of different Pt–Cu ratios on the morphology and size of nanoparticles. In addition, we further tuned the morphology of the Pt–Cu nanostructures by introducing bromide ions, eventually leading to the appearance of some tripod-like structures. Compared with dendritic Pt–Cu and commercial Pt black, these tripod-like Pt–Cu nanostructures exhibited higher electrocatalytic activity and CO tolerance for catalyzing methanol oxidation.

Online reviews provide consumers with information about products and services that may affect their purchasing decisions. As such, the customer attitude in the reviews play a important role to product sales. In this study, new sentiment prediction method is presented to enhance the forecasting accuracy by utilizing review dual-sentiments and employing non- linear manifold dynamics algorithm. Not only this method can extract and separate the polarity sentiment factors from the content of each online review, but also can remove the seasonality of products and mix the different types’ data. Extensive experimental results validate the effectiveness of our proposed method.

A sensitive “on-off” fluorescent protocol for thrombin detection is demonstrated. Firstly, thrombin aptamers which hybridize with labeled help DNA were immobilized on the surface of Ag@SiO2 nanoparticles (NPs). The silver core causes the label Cy5 to display strong metal-enhanced fluorescence. On addition of thrombin and graphene oxide (GO), thrombin (with its high affinity for the aptamers) displaces the Cy5-labeled help DNA which then binds to the surface of GO via π-stacking, causing fluorescence quenching of Cy5. The findings were used to design a thrombin assay that has a 0.05 nM detection limit and excellent selectivity. It was applied to the quantification of thrombin in spiked serum samples where is showed recoveries ranging from 97 % to 107 %, with relative standard deviations between 2.2 and 4.5 %.

The metal complex effect on the hydrogenation of chlorobenzene over a poly(N-vinyl-2-pyrrolidone)-stabilized platinum colloidal nanocatalyst (PVP-Pt) has been studied under 298 K and atmospheric pressure. The introduction of metal complexes and ligands exhibited great effect on the activity and the selectivity of the nanocatalyst. It is evidently manifested that the modification of metal complexes is not the simple sum of those of the metal central ions and the corresponding ligands.