Single-cell western blotting.

To measure cell-to-cell variation in protein-mediated functions, we developed an approach to conduct ∼10(3) concurrent single-cell western blots (scWesterns) in ∼4 h. A microscope slide supporting a 30-μm-thick photoactive polyacrylamide gel enables western blotting: settling of single cells into microwells, lysis in situ, gel electrophoresis, photoinitiated blotting to immobilize proteins and antibody probing. We applied this scWestern method to monitor single-cell differentiation of rat neural stem cells and responses to mitogen stimulation. The scWestern quantified target proteins even with off-target antibody binding, multiplexed to 11 protein targets per single cell with detection thresholds of <30,000 molecules, and supported analyses of low starting cell numbers (∼200) when integrated with FACS. The scWestern overcomes limitations of antibody fidelity and sensitivity in other single-cell protein analysis methods and constitutes a versatile tool for the study of complex cell populations at single-cell resolution

Platinum-Gallium (Pt-Ga) Intermetallic Alloys for Propane Dehydrogenation

Natural gas is a source of energy for the United States. The Center for Innovative Strategic Transformation of Alkane Resources (CISTAR) plans to use shale gas extracted from shale rock formations as a bridge fuel to replace coal and oil while the US transitions to renewable energy like solar and wind. After methane, the largest components in shale gas are light alkanes such as ethane and propane. These can be catalytically converted to olefins, which can be further reacted to produce fuels, for example. Olefins from alkanes can be accomplished by dehydrogenation by promoted platinum alloys. This study compares the structure and chemical properties of Pt-Ga alloys on silica (SiO2) and ceria (CeO2) supports to determine if the support plays an important role in this chemistry. The catalysts containing different Pt:Ga ratios were synthesized using incipient wetness impregnation. These catalysts were characterized by in situ X-ray diffraction (XRD) and X-ray adsorption spectroscopy (XAS) to determine if an alloy was formed, and if so, the structure of that alloy. Finally, the catalysts were tested in a fixed bed reactor, where it was found that the silica-supported Pt-Ga alloy has a selectivity of \u3e90% towards propylene. Understanding catalyst design can lead to higher catalytic conversion of substances and potentially an improved selectivity for the formation of preferred products. Pt-Ga on ceria is tested for comparison and there appears to behave differently from that on silica demonstrating the importance of the role of the support on these catalysts

A novel patient engagement platform using accessible text messages and calls (Epharmix): Feasibility study

BACKGROUND: Patient noncompliance with therapy, treatments, and appointments represents a significant barrier to improving health care delivery and reducing the cost of care. One method to improve therapeutic adherence is to improve feedback loops in getting clinically acute events and issues to the relevant clinical providers as necessary (ranging from detecting hypoglycemic events for patients with diabetes to notifying the provider when patients are out of medications). Patients often don\u27t know which information should prompt a call to their physician and proactive checks by the clinics themselves can be very resource intensive. We hypothesized that a two-way SMS system combined with a platform web service for providers would enable both high patient engagement but also the ability to detect relevant clinical alerts. OBJECTIVE: The objectives of this study are to develop a feasible two-way automated SMS/phone call + web service platform for patient-provider communication, and then study the feasibility and acceptability of the Epharmix platform. First, we report utilization rates over the course of the first 18 months of operation including total identified clinically significant events, and second, review results of patient user-satisfaction surveys for interventions for patients with diabetes, COPD, congestive heart failure, hypertension, surgical site infections, and breastfeeding difficulties. METHODS: To test this question, we developed a web service + SMS/phone infrastructure ( Epharmix ). Utilization results were measured based on the total number of text messages or calls sent and received, with percentage engagement defined as a patient responding to a text message at least once in a given week, including the number of clinically significant alerts generated. User satisfaction surveys were sent once per month over the 18 months to measure satisfaction with the system, frequency and degree of communication. Descriptive statistics were used to describe the above information. RESULTS: In total, 28,386 text messages and 24,017 calls were sent to 929 patients over 9 months. Patients responded to 80% to 90% of messages allowing the system to detect 1164 clinically significant events. Patients reported increased satisfaction and communication with their provider. Epharmix increased the number of patient-provider interactions to over 10 on average in any given month for patients with diabetes, COPD, congestive heart failure, hypertension, surgical site infections, and breastfeeding difficulties. CONCLUSIONS: Engaging high-risk patients remains a difficult process that may be improved through novel, digital health interventions. The Epharmix platform enables increased patient engagement with very low risk to improve clinical outcomes. We demonstrated that engagement among high-risk populations is possible when health care comes conveniently to where they are

Direct Laser Writing of Graphene Made from Chemical Vapor Deposition for Flexible, Integratable Micro-Supercapacitors with Ultrahigh Power Output

High‐performance yet flexible micro‐supercapacitors (MSCs) hold great promise as miniaturized power sources for increasing demand of integrated electronic devices. Herein, this study demonstrates a scalable fabrication of multilayered graphene‐based MSCs (MG‐MSCs), by direct laser writing (DLW) of stacked graphene films made from industry‐scale chemical vapor deposition (CVD). Combining the dry transfer of multilayered CVD graphene films, DLW allows a highly efficient fabrication of large‐areal MSCs with exceptional flexibility, diverse planar geometry, and capability of customer‐designed integration. The MG‐MSCs exhibit simultaneously ultrahigh energy density of 23 mWh cm−3 and power density of 1860 W cm−3 in an ionogel electrolyte. Notably, such MG‐MSCs demonstrate an outstanding flexible alternating current line‐filtering performance in poly(vinyl alcohol) (PVA)/H2SO4 hydrogel electrolyte, indicated by a phase angle of −76.2° at 120 Hz and a resistance–capacitance constant of 0.54 ms, due to the efficient ion transport coupled with the excellent electric conductance of the planar MG microelectrodes. MG–polyaniline (MG‐PANI) hybrid MSCs fabricated by DLW of MG‐PANI hybrid films show an optimized capacitance of 3.8 mF cm−2 in PVA/H2SO4 hydrogel electrolyte; an integrated device comprising MG‐MSCs line filtering, MG‐PANI MSCs, and pressure/gas sensors is demonstrated

Simultaneous Determination of 49 Antibiotics Residues in Pork by a Modified QuEChERS Method Based on Silanized Melamine Sponge Coupled with Ultra-high Performance Liquid Chromatography-Tandem Mass Spectrometry

Two new types of elastic porous silanized melamine sponges (MeS) were prepared by silylation reaction using octadecyltrichlorosilane (OTS) and N-[3-(trimethoxysilyl)propyl]ethylenediamine (ATS), which were respectively designated as OTS@MeS and ATS@MeS. The silanized sponges were used to develop a modified quick, easy, check, effective, rugged, and safe (QuEChERS) method that can quickly and efficiently separate interfering matrices from the extract through spontaneous solution infiltration and physical extrusion. In this study, an analytical method using the modified QuEChERS procedure combined with ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was established for the simultaneous determination of 49 antibiotic residues in pork. Samples were extracted with 10 mL of acetonitrile containing 0.1% acetic acid, and then salted out with 2.0 g of Na2SO4 and 0.5 g of NaCl. After centrifugation, a 1 mL aliquot of the supernatant was cleaned up with a mixture of OTS@MeS and ATS@MeS. The chromatographic separation was conducted on an Agilent ZORBAX Eclipse Plus C18 column with gradient elution using a mobile phase comprised of methanol and aqueous solution (methanol/water, 5:95, V/V) containing 0.1% formic acid and 5 mmol/L ammonium acetate. The qualitative and quantitative detection were performed by multiple-reaction monitoring (MRM) using an electrospray ionization source in the positive ion mode. The results showed that the correlation coefficients for all analytes were greater than 0.999. The matrix effects (ME) were in the range of −13.5%−10.9%. The limits of detection (LOQ) and quantitation (LOQ) were 0.1–10.0 and 0.3–33.3 μg/kg, respectively. The recoveries at three spiked levels ranged from 65.0% to 112.7%, with intra- and interday relative standard deviations (RSDs) of 0.3%–11.8% and 2.4%–18.4%, respectively. The developed method was simple, rapid, highly sensitive and accurate, and could be used for the efficient and rapid determination of the 49 antibiotics residues in pork

Evidence-Based Annotation of Gene Function in Shewanella oneidensis MR-1 Using Genome-Wide Fitness Profiling across 121 Conditions

Most genes in bacteria are experimentally uncharacterized and cannot be annotated with a specific function. Given the great diversity of bacteria and the ease of genome sequencing, high-throughput approaches to identify gene function experimentally are needed. Here, we use pools of tagged transposon mutants in the metal-reducing bacterium Shewanella oneidensis MR-1 to probe the mutant fitness of 3,355 genes in 121 diverse conditions including different growth substrates, alternative electron acceptors, stresses, and motility. We find that 2,350 genes have a pattern of fitness that is significantly different from random and 1,230 of these genes (37% of our total assayed genes) have enough signal to show strong biological correlations. We find that genes in all functional categories have phenotypes, including hundreds of hypotheticals, and that potentially redundant genes (over 50% amino acid identity to another gene in the genome) are also likely to have distinct phenotypes. Using fitness patterns, we were able to propose specific molecular functions for 40 genes or operons that lacked specific annotations or had incomplete annotations. In one example, we demonstrate that the previously hypothetical gene SO_3749 encodes a functional acetylornithine deacetylase, thus filling a missing step in S. oneidensis metabolism. Additionally, we demonstrate that the orphan histidine kinase SO_2742 and orphan response regulator SO_2648 form a signal transduction pathway that activates expression of acetyl-CoA synthase and is required for S. oneidensis to grow on acetate as a carbon source. Lastly, we demonstrate that gene expression and mutant fitness are poorly correlated and that mutant fitness generates more confident predictions of gene function than does gene expression. The approach described here can be applied generally to create large-scale gene-phenotype maps for evidence-based annotation of gene function in prokaryotes

Conversion of Shale Gas with Supported Metal Catalysts

As shale gas exploitation has been developed, production of shale gas in the US has rapidly increased during the last decade. This has motivated the development of techniques to covert shale gas components (mainly C1 to C3) to liquid fuels by catalytic conversion. The main goal of the dissertation is to study the geometric and electronic structures of the metal catalysts, which are crucial for understanding the structure-property relationship. The first project studies bimetallic Pt-Bi catalyst for non-oxidative coupling of methane. In a recent publication published in ACS catalysis, Pt-Bi/ZSM-5 catalyst has been shown to stably convert methane into C2 for 8 hours under non-oxidative conditions. In this manuscript, structure of the Pt-Bi/ZSM-5 was shown with HAADF imaging, synchrotron XAS and XRD. A new surface cubic Pt3Bi phase on Pt nanoparticles with Pt-Bi bond distance of 2.80 ˚A was formed. Formation of noble metal intermetallic alloys such as Pt3M may be the clue for non-oxidative conversion of methane. The second and third project highlight strong metal-support interaction catalysts for propane dehydrogenation. Chemisorption showed partial coverage of the SMSI oxides on the surface of the nanoparticles. In situX-ray absorption near edge (XANES), resonant inelastic X-ray scattering (RIXS), X-ray photoelectron spectroscopy (XPS) have shown that little electronic effect on the metal nanoparticles. The catalyst activity per mol of metal decreased due to the partial coverage of the SMSI oxides on the surface of the catalysts. The catalysts, however, had higher selectivity due to smaller ensembles inhibiting hydrogenolysis. In the fourth project Pt-P catalyst was investigated to understand the promoting effect of P.Pt-P catalysts had much higher selectivity for propane dehydrogenation (\u3e95%). These give two types of catalysts, a PtP2-rich surface on Pt core and full PtP2 ordered structure, which were confirmed by scanning transmission electron microscopy (STEM), and in situ methods of EXAFS, synchrotron XRD, XPS, and Resonant Inelastic X-ray Spectroscopy (RIXS). The PtP2 structure has isolated Pt atoms separated by P2atoms. In addition XANES, XPS and RIXS indicate a strong electronic modification in the energy of the valence orbitals. It can be concluded from the Pt-Bi catalyst that intermetallic alloys might be selective for NOCM. Therefore, promoters with higher reduction temperature, such as Mn and Cr, should be used to have stable catalysts at high temperature. Moreover, both Pt-Bi and Pt/CeO2suggest that selective catalysts for propane dehydrogenation and NOCM may have some correlation. Further studies would be conducted to understand the correlation between the two reactions

Multidistribution Center Location Based on Real-Parameter Quantum Evolutionary Clustering Algorithm

To determine the multidistribution center location and the distribution scope of the distribution center with high efficiency, the real-parameter quantum-inspired evolutionary clustering algorithm (RQECA) is proposed. RQECA is applied to choose multidistribution center location on the basis of the conventional fuzzy C-means clustering algorithm (FCM). The combination of the real-parameter quantum-inspired evolutionary algorithm (RQIEA) and FCM can overcome the local search defect of FCM and make the optimization result independent of the choice of initial values. The comparison of FCM, clustering based on simulated annealing genetic algorithm (CSAGA), and RQECA indicates that RQECA has the same good convergence as CSAGA, but the search efficiency of RQECA is better than that of CSAGA. Therefore, RQECA is more efficient to solve the multidistribution center location problem