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Heterogeneous catalysts coupled with non-thermal plasma are known to achieve higher reaction yields, but disentangling the gain achieved by the individual components remains challenging. Here, NO is produced from N2 and O2 in a plasma-catalytic reactor at conditions at which neither catalyst nor plasma alone is productive, providing unambiguous evidence of plasma-catalyst synergy.

Hypothesis: The purpose of this study was to determine the incidence of clinically significant postoperative stiffness after arthroscopic rotator cuff repair and its resolution. The study also sought to determine clinical and surgical factors that may be associated with increased rates of postoperative stiffness. Methods: We conducted a level III retrospective review of a consecutive series of arthroscopic rotator cuff repairs. During a 5-year period, the senior author (C.J.R.) performed 150 arthroscopic rotator cuff repairs at our institution. Demographic data, comorbid medical conditions, descriptions of rotator cuff tears (including size and level of retraction), and concomitant surgical procedures were evaluated on their correlation with stiffness. All office visits were reviewed to determine preoperative and postoperative motion. Patients were followed up at 1 week, 3 weeks, 6-8 weeks, 3 months, about 6 months, and 1 year postoperatively. Results: In our analysis of tear types, we were unable to associate stiffness with the type of tear, the tendon torn, or the number of tendons torn or with whether the tendons were retracted. However, we were able to associate female sex, workers' compensation insurance, and a concomitant biceps procedure with stiffness at several time points. The incidence of stiffness was highest at 12 weeks, with 7.3% of patients presenting with stiffness. The rate of stiffness decreased with continued follow-up. Stiffness was found in 3.3% of patients at 16-24 weeks and in 1.6% of patients at 1 year. Conclusions: Prolonged physical therapy will result in resolution of stiffness in the vast majority of cases, often obviating the return to the operating room for capsular release and lysis of adhesions or mobilization under anesthesia.

To model hepatocyte function accurately in vitro, it is necessary to generate and maintain a polarized epithelium. Here, the authors describe a protocol to generate polarized human pluripotent stem cell-derived hepatocyte-like cells (HLCs) to model enteric virus production and drug secretion in vitro.

Abstract Many RNA viruses have genetically diverse populations in a single host. Important biological characteristics may be related to the levels of diversity, including adaptability, host specificity, and host range. Shifting the virus between hosts might result in a change in the levels of diversity associated with the new host. The level of genetic diversity for these viruses is related to host, vector and virus interactions, and understanding these interactions may facilitate the prediction and prevention of emerging viral diseases. It is known that luteoviruses have a very specific interaction with aphid vectors. Previous studies suggested that there may be a tradeoff effect between the viral adaptation and aphid transmission when Soybean dwarf virus (SbDV) was transmitted into new plant hosts by aphid vectors. In this study, virus titers in different aphid vectors and the levels of population diversity of SbDV in different plant hosts were examined during multiple sequential aphid transmission assays. The diversity of SbDV populations revealed biases for particular types of substitutions and for regions of the genome that may incur mutations among different hosts. Our results suggest that the selection on SbDV in soybean was probably leading to reduced efficiency of virus recognition in the aphid which would inhibit movement of SbDV through vector tissues known to regulate the specificity relationship between aphid and virus in many systems.

Sleep disturbance is a common and disruptive symptom of neurodegenerative diseases such as Alzheimer's and Huntington's disease (HD). In HD patients, sleep fragmentation appears at an early stage of disease, although features of the earliest sleep abnormalities in presymptomatic HD are not fully established. Here we used novel automated analysis of quantitative electroencephalography to study transitions between wake and non-rapid eye movement sleep in a sheep model of presymptomatic HD. We found that while the number of transitions between sleep and wake were similar in normal and HD sheep, the dynamics of transitions from sleep-to-wake differed markedly between genotypes. Rather than the gradual changes in EEG power that occurs during transitioning from sleep-to-wake in normal sheep, transition into wake was abrupt in HD sheep. Furthermore, transitions to wake in normal sheep were preceded by a significant reduction in slow wave power, whereas in HD sheep this prior reduction in slow wave power was far less pronounced. This suggests an impaired ability to prepare for waking in HD sheep. The abruptness of awakenings may also have potential to disrupt sleep-dependent processes if they are interrupted in an untimely and disjointed manner. We propose that not only could these abnormal dynamics of sleep transitions be useful as an early biomarker of HD, but also that our novel methodology would be useful for studying transition dynamics in other sleep disorders.

Zika virus (ZIKV) is a re-emerging flavivirus that has caused large-scale epidemics. Infection during pregnancy can lead to neurologic developmental abnormalities in children. There is no approved vaccine or therapy for ZIKV. To uncover cellular pathways required for ZIKV that can be therapeutically targeted, we transcriptionally upregulated all known human coding genes with an engineered CRISPR–Cas9 activation complex in human fibroblasts deficient in interferon (IFN) signaling. We identified Ras homolog family member V (RhoV) and WW domain-containing transcription regulator 1 (WWTR1) as proviral factors, and found them to play important roles during early ZIKV infection in A549 cells. We then focused on RhoV, a Rho GTPase with atypical terminal sequences and membrane association, and validated its proviral effects on ZIKV infection and virion production in SNB-19 cells. We found that RhoV promotes infection of some flaviviruses and acts at the step of viral entry. Furthermore, RhoV proviral effects depend on the complete GTPase cycle. By depleting Rho GTPases and related proteins, we identified RhoB and Pak1 as additional proviral factors. Taken together, these results highlight the positive role of RhoV in ZIKV infection and confirm CRISPR activation as a relevant method to identify novel host–pathogen interactions.

Mixed halide hybrid perovskites possess tunable band gaps, however, under illumination they undergo phase separation. Using spectroscopic measurements and theoretical modelling, Draguta and Sharia et al. quantitatively rationalize the microscopic processes that occur during phase separation.

ABSTRACT Rathayibacter toxicus is a species of Gram-positive, corynetoxin-producing bacteria that causes annual ryegrass toxicity, a disease often fatal to grazing animals. A phylogenomic approach was employed to model the evolution of R. toxicus to explain the low genetic diversity observed among isolates collected during a 30-year period of sampling in three regions of Australia, gain insight into the taxonomy of Rathayibacter, and provide a framework for studying these bacteria. Analyses of a data set of more than 100 sequenced Rathayibacter genomes indicated that Rathayibacter forms nine species-level groups. R. toxicus is the most genetically distant, and evidence suggested that this species experienced a dramatic event in its evolution. Its genome is significantly reduced in size but is colinear to those of sister species. Moreover, R. toxicus has low intergroup genomic diversity and almost no intragroup genomic diversity between ecologically separated isolates. R. toxicus is the only species of the genus that encodes a clustered regularly interspaced short palindromic repeat (CRISPR) locus and that is known to host a bacteriophage parasite. The spacers, which represent a chronological history of infections, were characterized for information on past events. We propose a three-stage process that emphasizes the importance of the bacteriophage and CRISPR in the genome reduction and low genetic diversity of the R. toxicus species. IMPORTANCE Rathayibacter toxicus is a toxin-producing species found in Australia and is often fatal to grazing animals. The threat of introduction of the species into the United States led to its inclusion in the Federal Select Agent Program, which makes R. toxicus a highly regulated species. This work provides novel insights into the evolution of R. toxicus. R. toxicus is the only species in the genus to have acquired a CRISPR adaptive immune system to protect against bacteriophages. Results suggest that coexistence with the bacteriophage NCPPB3778 led to the massive shrinkage of the R. toxicus genome, species divergence, and the maintenance of low genetic diversity in extant bacterial groups. This work contributes to an understanding of the evolution and ecology of an agriculturally important species of bacteria.

The host factor and interferon (IFN)-stimulated gene (ISG) product, zinc-finger antiviral protein (ZAP), inhibits a number of diverse viruses by usurping and intersecting with multiple cellular pathways. To elucidate its antiviral mechanism, we perform a loss-of-function genome-wide RNAi screen to identify cellular cofactors required for ZAP antiviral activity against the prototype alphavirus, Sindbis virus (SINV). In order to exclude off-target effects, we carry out stringent confirmatory assays to verify the top hits. Important ZAP-liaising partners identified include proteins involved in membrane ion permeability, type I IFN signaling, and post-translational protein modification. The factor contributing most to the antiviral function of ZAP is TRIM25, an E3 ubiquitin and ISG15 ligase. We demonstrate here that TRIM25 interacts with ZAP through the SPRY domain, and TRIM25 mutants lacking the RING or coiled coil domain fail to stimulate ZAP's antiviral activity, suggesting that both TRIM25 ligase activity and its ability to form oligomers are critical for its cofactor function. TRIM25 increases the modification of both the short and long ZAP isoforms by K48- and K63-linked polyubiquitin, although ubiquitination of ZAP does not directly affect its antiviral activity. However, TRIM25 is critical for ZAP's ability to inhibit translation of the incoming SINV genome. Taken together, these data uncover TRIM25 as a bona fide ZAP cofactor that leads to increased ZAP modification enhancing its translational inhibition activity.

Rathayibacter toxicus is a forage grass associated Gram-positive bacterium of major concern to food safety and agriculture. This species is listed by USDA-APHIS as a plant pathogen select agent because it produces a tunicamycin-like toxin that is lethal to livestock and may be vectored by nematode species native to the U.S. The complete genomes of two strains of R. toxicus, including the type strain FH-79, were sequenced and analyzed in comparison with all available, complete R. toxicus genomes. Genome sizes ranged from 2,343,780 to 2,394,755 nucleotides, with 2079 to 2137 predicted open reading frames; all four strains showed remarkable synteny over nearly the entire genome, with only a small transposed region. A cluster of genes with similarity to the tunicamycin biosynthetic cluster from Streptomyces chartreusis was identified. The tunicamycin gene cluster (TGC) in R. toxicus contained 14 genes in two transcriptional units, with all of the functional elements for tunicamycin biosynthesis present. The TGC had a significantly lower GC content (52%) than the rest of the genome (61.5%), suggesting that the TGC may have originated from a horizontal transfer event. Further analysis indicated numerous remnants of other potential horizontal transfer events are present in the genome. In addition to the TGC, genes potentially associated with carotenoid and exopolysaccharide production, bacteriocins and secondary metabolites were identified. A CRISPR array is evident. There were relatively few plant-associated cell-wall hydrolyzing enzymes, but there were numerous secreted serine proteases that share sequence homology to the pathogenicity-associated protein Pat-1 of Clavibacter michiganensis. Overall, the genome provides clear insight into the possible mechanisms for toxin production in R. toxicus, providing a basis for future genetic approaches.

Bleeding, is one of the most common postoperative complications after palatoplasty in patients with cleft palate. The purpose of this article is to present our experience using a Foley catheter balloon for perioperative palatoplasty bleeding management. A retrospective data analysis was performed for all cases of palatoplasty performed by the author from 1999 to 2012 that experienced postoperative hemorrhage. These patients were managed by utilizing posterior nasal compression with a Foley catheter balloon for the period 2006 to 2012. We have compared two methods (we use before 2006 the reoperative method) with regards to the later development of palatal fistulae and cases with reoperative hemostasis. The study found no statistically significant differences between the conventional and proposed method in relation to the development of palatal fistulae (P=0.7). These findings suggest that nasal packing using a Foley balloon is a safe and effective method for postoperative bleeding control after palatoplasty in patients with cleft palate.

To encourage sustainable engineering practices, departments of transportation are interested in reusing winter maintenance truck wash water as part of their brine production and future road application. Traffic-related metals in the wash water, however, could limit this option. The objective of this work was to conduct a pilot scale evaluation of heavy metal (copper, zinc, iron, and lead) removal in a filtration unit (maximum flow rate of 45 L/minute) containing proprietary (MAR Systems Sorbster®) media. Three different trials were conducted and approximately 10,000 L of wash water collected from a winter maintenance facility in Ohio was treated with the pilot unit. Lab studies were also performed on six wash-water samples from multiple facilities to assess particle size removal and estimate settling time as a potential removal mechanism during wash-water storage. Pilot unit total metal removal efficiencies were 79%, 77%, 63%, and 94% for copper, zinc, iron, and lead, respectively. Particle settling calculation estimates for copper and zinc show that 10 hours in storage can also effectively reduce heavy metal concentrations in winter maintenance wash water in excess of 70%. These pilot scale results show promise for reducing heavy metal concentrations to an acceptable level for reuse.

The Ohio Department of Transportation has more than 60 facilities without sewer access generating approximately 19 million gallons of winter maintenance wash water. Off-site disposal is costly, creating the need for sustainable management strategies. The objective of this study was to conduct an exploratory feasibility analysis to assess wash water disposal and potential reuse as brine. Based on a comprehensive literature review and relevant environmental chemistry, a sampling protocol consisting of 31 water quality constituents was utilized for monthly sampling at three geographically distinct Ohio Department of Transportation garages during the winter of 2012. Results were compared to local disposal and reuse guidance limits. Three constituents, including a maximum copper concentration of 858 ppb, exceeded disposal limits, and many constituents also failed to meet reuse limits. Some concentrations were orders of magnitude higher than reuse limits and suggest pre-treatment would be necessary if wash water were reused as brine. These water quality results, in conjunction with copper chemical equilibrium modeling, show pH and dissolved carbon both significantly impact the total dissolved copper concentration and should be measured to assess reuse potential. The sampling protocol and specific obstacles highlighted in this paper aid in the future development of sustainable wash water management strategies.

Solvent-free, single-ion conducting electrolytes are sought after for use in electrochemical energy storage devices. Here, we investigate the ionic conductivity and how this property is influenced by segmental mobility and conducting ion number in crosslinked single-ion conducting polyether-based electrolytes with varying tethered anion and counter-cation types. Crosslinked electrolytes are prepared by the polymerization of poly(ethylene glycol) diacrylate (PEGDA), poly(ethylene glycol) methyl ether acrylate, and ionic monomers. The ionic conductivity of the electrolytes is measured and interpreted in the context of differential scanning calorimetry and Raman spectroscopy measurements. A lithiated crosslinked electrolyte prepared with PEG31DA and (4-styrenesulfonyl)(trifluoromethanesulfonyl)imide (STFSI) monomers is found to have a lithium ion conductivity of 3.2 × 10−6 and 1.8 × 10−5 S/cm at 55 and 100 °C, respectively. The percentage of unpaired anions for this electrolyte was estimated at about 23% via Raman spectroscopy. Despite the large variances in metal cation–STFSI binding energies as predicted via density functional theory (DFT) and large variations in ionic conductivity, STFSI-based crosslinked electrolytes with the same charge density and varying cations (Li, Na, K, Mg, and Ca) were estimated to all have unpaired anion populations in the range of 19 to 29%.

The original version of this Article contained an error in the spelling of the author Joseph S. Manser, which was incorrectly given as Joseph M. Manser. This has now been corrected in both the PDF and HTML versions of the Article.

The hepatitis C virus (HCV) p7 protein is required for infectious virus production via its role in assembly and ion channel activity. Although NMR structures of p7 have been reported, the location of secondary structural elements and orientation of the p7 transmembrane domains differ among models. Furthermore, the p7 structure-function relationship remains unclear. Here, extensive mutagenesis, coupled with infectious virus production phenotyping and molecular modeling, demonstrates that the N-terminal helical region plays a previously underappreciated yet critical functional role, especially with respect to E2/p7 cleavage efficiency. Interrogation of specific N-terminal helix residues identified as having p7-specific defects and predicted to point toward the channel pore, in a context of independent E2/p7 cleavage, further supports p7 as a structurally plastic, minimalist ion channel. Together, our findings indicate that the p7 N-terminal helical region is critical for E2/p7 processing, protein-protein interactions, ion channel activity, and infectious HCV production.

Virus-induced gene silencing (VIGS) is used to analyze gene function in dicotyledonous plants but less so in monocotyledonous plants (particularly rice and corn), partially due to the limited number of virus expression vectors available. Here, we report the cloning and modification for VIGS of a virus from Festuca arundinacea Schreb. (tall fescue) that caused systemic mosaic symptoms on barley, rice, and a specific cultivar of maize (Va35) under greenhouse conditions. Through sequencing, the virus was determined to be a strain of Brome mosaic virus (BMV). The virus was named F-BMV (F for Festuca), and genetic determinants that controlled the systemic infection of rice were mapped to RNAs 1 and 2 of the tripartite genome. cDNA from RNA 3 of the Russian strain of BMV (R-BMV) was modified to accept inserts from foreign genes. Coinoculation of RNAs 1 and 2 from F-BMV and RNA 3 from R-BMV expressing a portion of a plant gene to leaves of barley, rice, and maize plants resulted in visual silencing-like phenotypes. The visual phenotypes were correlated with decreased target host transcript levels in the corresponding leaves. The VIGS visual phenotype varied from maintained during silencing of actin 1 transcript expression to transient with incomplete penetration through affected tissue during silencing of phytoene desaturase expression. F-BMV RNA 3 was modified to allow greater accumulation of virus while minimizing virus pathogenicity. The modified vector C-BMVA/G (C for chimeric) was shown to be useful for VIGS. These BMV vectors will be useful for analysis of gene function in rice and maize for which no VIGS system is reported.

The increasing prevalence of quinolone-resistant Neisseria gonorrhoeae (QRNG) in the United States is a cause for concern. Detecting resistance is complicated by the widespread use of molecular tests that do not provide isolates for susceptibility testing. The Michigan Department of Community Health developed a sentinel surveillance program to detect antimicrobial drug resistance in N. gonorrhoeae. Sentinel surveillance from 11 laboratories submitted 1,122 isolates for antimicrobial drug susceptibility testing and detected 2 clusters of QRNG from January 2003 to September 2004. These clusters were epidemiologically distinct: one involved young, heterosexual youth, and the other involved older men who have sex with men. This finding led to changes in local treatment recommendations that limited spread of resistant strains. Development of the sentinel program, collection of data, and epidemiologic analysis of the clusters are discussed.

The integration of local knowledge and science represents an opportunity to enhance the understanding of interrelations among climate, hydrology, and socioeconomic systems while providing mutual benefits to scientists and rural communities. Insight from rural Alaskans helped to identify a social-ecological threshold used to model potential driftwood harvest from the Yukon River. Information from residents of Tanana, Alaska, was combined with scientific data to model driftwood harvest rates. Modeling results estimated that between 1980 and 2010, hydrologic factors alone were responsible for a 29% decrease in the annual wood harvest, which approximately balanced a 23% reduction in wood demand because of a decline in number of households. The community's installation of wood-fired boilers in 2007 created a threshold increase (76%) in wood demand that is not met by driftwood harvest. Modeling analyses of numerous climatic scenarios illustrated that increases in hydrologic variability would decrease the reliability of future driftwood harvest. Economic analyses demonstrated that increased climatic variability could have serious economic consequences for subsistence users while demanding more of their time. Lost time is important because it reduces their availability for performing other subsistence activities and learning to adapt to climate-related challenges. Our research may benefit communities by providing a tool that can be used to predict the timing and duration of driftwood runs. Information gathered from discussions with local stakeholders provided critical information for model development and thus provided a better understanding of regional social-ecological dynamics. Our research also illustrates the potential for regional-scale adaptations to limit the social-ecological impacts of environmental change, while providing economic opportunities and energy independence that reduce their vulnerability to variations in climate.