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In this study, the correlation between the composition, hydrogen/carbon (H/C) ratio, and hydrogen/oxygen (H/O) ratio of a used cooking oil (UCO) and the specific emissions embodied in the derived hydroprocessed esters and fatty acids synthetic paraffinic kerosene (HEFA‐SPK) is investigated. It is shown that HEFA‐SPK produced from UCOs with low concentrations of C18:1, high concentrations of C18:2, and low H/C ratios utilize less energy and more hydrogen during Fuel Production. Hence, HEFA‐SPK produced from such UCOs will embody higher gCO2e for fossil hydrogen utilization scenarios, and lower gCO2e for green hydrogen utilization scenarios compared with other UCOs. Conversely, it is shown that HEFA‐SPK produced from UCOs with high concentrations of C18:1, low concentrations of C18:2 and high H/C ratios utilize more energy and less hydrogen during Fuel Production. Hence, HEFA‐SPK derived from such UCOs will embody lower gCO2e for fossil hydrogen utilization scenarios, and higher gCO2e for green hydrogen utilization scenarios compared with other UCOs. Monte Carlo simulation gives the emissions embodied in Fuel Production a 95% confidence interval for all UCO‐derived HEFA‐SPK, showing a similar uncertainty for all compositions. A maximum of +1.1 gCO2e/MJSAF and −1.0 gCO2e/MJSAF is obtained for the upper and lower bounds respectively for the emissions embodied during HEFA‐SPK production from UCO. The application of the correlations founded in this study allows for the prediction of the specific emissions embodied in the feedstock‐to‐fuel conversion of any UCO, providing the C18:1 concentration, C18:2 concentration, H/C ratio and H/O ratio of the UCO is known. [ABSTRACT FROM AUTHOR]

A high-throughput screen to discover inhibitors of p97 ATPase activity identified an indole amide that bound to an allosteric site of the protein. Medicinal chemistry optimization led to improvements in potency and solubility. Indole amide 3 represents a novel uncompetitive inhibitor with excellent physical and pharmaceutical properties that can be used as a starting point for drug discovery efforts.

Actin filament networks assemble on cellular membranes in response to signals that locally activate neural Wiskott-Aldrich-syndrome protein (N-WASP) and the Arp2/3 complex. An inactive conformation of N-WASP is stabilized by intramolecular contacts between the GTPase binding domain (GBD) and the C helix of the verprolin-homology, connector-helix, acidic motif (VCA) segment. Multiple SH3 domain-containing adapter proteins can bind and possibly activate N-WASP, but it remains unclear how such binding events relieve autoinhibition to unmask the VCA segment and activate the Arp2/3 complex. Here, we have used purified components to reconstitute a signaling cascade driven by membrane-localized Src homology 3 (SH3) adapters and N-WASP, resulting in the assembly of dynamic actin networks. Among six SH3 adapters tested, Nck was the most potent activator of N-WASP-driven actin assembly. We identify within Nck a previously unrecognized activation motif in a linker between the first two SH3 domains. This linker sequence, reminiscent of bacterial virulence factors, directly engages the N-WASP GBD and competes with VCA binding. Our results suggest that animals, like pathogenic bacteria, have evolved peptide motifs that allosterically activate N-WASP, leading to localized actin nucleation on cellular membranes.

Lean construction (LC) and building information modeling (BIM) are two of the prominent concepts challenging the traditional practices in construction management. Small and medium-sized enterprises (SMEs) often constitute the largest group in construction supply chains. Increasing BIM and LC adoption amongst SMEs is a key condition for achieving the transformation of the construction industry through BIM and LC. The paper presents a systematic literature review of the adoption of (i) LC, (ii) BIM, and (iii) both LC and BIM in SMEs to evaluate the current literature, and 114 papers were included in the review. The bibliographic and content characteristics of the literature were discussed in detail. It was found that despite the importance of SMEs, the current LC and BIM literature falls short in terms of both number of publications and content of publications. The paper concludes with some generic suggestions for future research and action. Key words: state-of-the-art review, building information modeling (BIM), lean construction, small and medium-sized enterprises (SMEs), construction management. La construction frugale (CF) et la modelisation des donnees du batiment (<< BIM >>) sont deux des principaux concepts qui remettent en question les pratiques traditionnelles de gestion de la construction. Les petites et moyennes entreprises (PME) constituent souvent le groupe le plus important dans les chaines d'approvisionnement de la construction. Promouvoir l'adoption de la BIM et de la CF dans les PME est une condition essentielle a la realisation de la transformation de l'industrie de la construction grace a la BIM et la CF. Cet article presente une revue systematique de la litterature portant sur l'adoption (i) de la CF, (ii) de la BIM et (iii) de ces deux concepts dans les PME afin d'evaluer la litterature actuelle. Notre revue compte 114 articles. Les caracteristiques bibliographiques et le contenu de la litterature ont ete analyses en detail. On a constate que, malgre l'importance des PME, la litterature actuelle au sujet de la CF et la BIM est insuffisante tant en termes de nombre de publications que de contenu. Le document se termine par quelques suggestions generales quant a la recherche et a l'action futures. [Traduit par la Redaction] Mots-cles : examen approfondi, modelisation des donnees du batiment (BIM), construction frugale (CF), petite ou moyenne entreprise (PME), gestion de la construction., 1. Introduction The construction industry in most countries is categorised by a small number of larger companies and a very large number of small and medium-sized enterprises (SMEs). In many [...]

The retinoblastoma binding protein KDM5A removes methyl marks from lysine 4 of histone H3 (H3K4). Misregulation of KDM5A contributes to the pathogenesis of lung and gastric cancers. In addition to its catalytic jumonji C domain, KDM5A contains three PHD reader domains, commonly recognized as chromatin recruitment modules. It is unknown whether any of these domains in KDM5A have functions beyond recruitment and whether they regulate the catalytic activity of the demethylase. Here using biochemical and nuclear magnetic resonance (NMR)-based structural studies, we show that the PHD1 preferentially recognizes unmethylated H3K4 histone tail, product of KDM5A-mediated demethylation of tri-methylated H3K4 (H3K4me3). Binding of unmodified H3 peptide to the PHD1 stimulates catalytic domain-mediated removal of methyl marks from H3K4me3 peptide and nucleosome substrates. This positive-feedback mechanism--enabled by the functional coupling between a reader and a catalytic domain in KDM5A--suggests a model for the spread of demethylation on chromatin.

Hsp90 is a conformationally dynamic molecular chaperone known to promote the folding and activation of a broad array of protein substrates ("clients"). Hsp90 is believed to preferentially interact with partially folded substrates, and it has been hypothesized that the chaperone can significantly alter substrate structure as a mechanism to alter the substrate functional state. However, critically testing the mechanism of substrate recognition and remodeling by Hsp90 has been challenging. Using a partially folded protein as a model system, we find that the bacterial Hsp90 adapts its conformation to the substrate, forming a binding site that spans the middle and C-terminal domains of the chaperone. Cross-linking and NMR measurements indicate that Hsp90 binds to a large partially folded region of the substrate and significantly alters both its local and long-range structure. These findings implicate Hsp90's conformational dynamics in its ability to bind and remodel partially folded proteins. Moreover, native-state hydrogen exchange indicates that Hsp90 can also interact with partially folded states only transiently populated from within a thermodynamically stable, native-state ensemble. These results suggest a general mechanism by which Hsp90 can recognize and remodel native proteins by binding and remodeling partially folded states that are transiently sampled from within the native ensemble.

BackgroundMild brain hypothermia (32°-34°C) after human neonatal asphyxia improves neurodevelopmental outcomes. Astrocytes but not neurons have pyruvate carboxylase and an acetate uptake transporter. C nuclear magnetic resonance spectroscopy of rodent brain extracts after administering [1-C]glucose and [1,2-C]acetate can distinguish metabolic differences between glia and neurons, and tricarboxylic acid cycle entry via pyruvate dehydrogenase and pyruvate carboxylase.MethodsNeonatal rat cerebrocortical slices receiving a C-acetate/glucose mixture underwent a 45-min asphyxia simulation via oxygen-glucose-deprivation followed by 6 h of recovery. Protocols in three groups of N=3 experiments were identical except for temperature management. The three temperature groups were: normothermia (37°C), hypothermia (32°C for 3.75 h beginning at oxygen--glucose deprivation start), and delayed hypothermia (32°C for 3.75 h, beginning 15 min after oxygen-glucose deprivation start). Multivariate analysis of nuclear magnetic resonance metabolite quantifications included principal component analyses and the L1-penalized regularized regression algorithm known as the least absolute shrinkage and selection operator.ResultsThe most significant metabolite difference (P

BACKGROUND: Mild brain hypothermia (31-34 °C) after neonatal hypoxia-ischemia (HI) improves neurodevelopmental outcomes in human and animal neonates. Using an asphyxia model with neonatal mice treated with mild hypothermia after HI, we investigated whether (1)H nuclear magnetic resonance (NMR) metabolomics of brain extracts could suggest biomarkers and distinguish different treatments and outcome groups. METHODS: At postnatal day 7 (P7), CD1 mice underwent right carotid artery occlusion, 30 min of HI (8% oxygen), and 3.5 h of either hypothermia (31 °C) or normothermia (37 °C). Whole brains were frozen immediately after HI, immediately after 3.5 h of hypothermia or normothermia treatments, and 24 h later. Perchloric acid extractions of 36 metabolites were quantified by 900 MHz (1)H NMR spectroscopy. Multivariate analyses included principal component analyses (PCA) and a novel regression algorithm. Histological injury was quantified after HI at 5 d. RESULTS: PCA scores plots separated normothermia/HI animals from hypothermia/HI and control animals, but more data are required for multivariate models to be predictive. Loadings plots identified 11 significant metabolites, whereas the regression algorithm identified 6. Histological injury scores were significantly reduced by hypothermia. CONCLUSION: Different treatment and outcome groups are identifiable by (1)H NMR metabolomics in a neonatal mouse model of mild hypothermia treatment of HI.

Human clinical trials using 72 hours of mild hypothermia (32°C-34°C) after neonatal asphyxia have found substantially improved neurologic outcomes. As temperature changes differently modulate numerous metabolite fluxes and concentrations, we hypothesized that (1)H/(31)P nuclear magnetic resonance (NMR) spectroscopy of intracellular metabolites can distinguish different insults, treatments, and recovery stages. Three groups of superfused neonatal rat brain slices underwent 45 minutes oxygen-glucose deprivation (OGD) and then were: treated for 3 hours with mild hypothermia (32°C) that began with OGD, or similarly treated with hypothermia after a 15-minute delay, or not treated (normothermic control group, 37°C). Hypothermia was followed by 3 hours of normothermic recovery. Slices collected at different predetermined times were processed, respectively, for 14.1 Tesla NMR analysis, enzyme-linked immunosorbent assay (ELISA) cell-death quantification, and superoxide production. Forty-nine NMR-observable metabolites underwent a multivariate analysis. Separated clustering in scores plots was found for treatment and outcome groups. Final ATP (adenosine triphosphate) levels, severely decreased at normothermia, were restored equally by immediate and delayed hypothermia. Cell death was decreased by immediate hypothermia, but was equally substantially greater with normothermia and delayed hypothermia. Potentially important biomarkers in the (1)H spectra included PCr-(1)H (phosphocreatine in the (1)H spectrum), ATP-(1)H (adenosine triphosphate in the (1)H spectrum), and ADP-(1)H (adenosine diphosphate in the (1)H spectrum). The findings suggest a potential role for metabolomic monitoring during therapeutic hypothermia.

Newly generated actin free barbed ends at the front of motile cells provide sites for actin filament assembly driving membrane protrusion. Growth factors induce a rapid biphasic increase in actin free barbed ends, and we found both phases absent in fibroblasts lacking H(+) efflux by the Na-H exchanger NHE1. The first phase is restored by expression of mutant cofilin-H133A but not unphosphorylated cofilin-S3A. Constant pH molecular dynamics simulations and nuclear magnetic resonance (NMR) reveal pH-sensitive structural changes in the cofilin C-terminal filamentous actin binding site dependent on His133. However, cofilin-H133A retains pH-sensitive changes in NMR spectra and severing activity in vitro, which suggests that it has a more complex behavior in cells. Cofilin activity is inhibited by phosphoinositide binding, and we found that phosphoinositide binding is pH-dependent for wild-type cofilin, with decreased binding at a higher pH. In contrast, phosphoinositide binding by cofilin-H133A is attenuated and pH insensitive. These data suggest a molecular mechanism whereby cofilin acts as a pH sensor to mediate a pH-dependent actin filament dynamics.