Structural insights into peptide self-assembly using photo-induced crosslinking experiments and discontinuous molecular dynamics

Determining the structure of the (oligomeric) intermediates that form during the self-assembly of amyloidogenic peptides is challenging because of their heterogeneous and dynamic nature. Thus, there is need for methodology to analyze the underlying molecular structure of these transient species. In this work, a combination of fluorescence quenching, photo-induced crosslinking (PIC) and molecular dynamics simulation was used to study the assembly of a synthetic amyloid-forming peptide, Aβ_16-22. A PIC amino acid containing a trifluormethyldiazirine (TFMD) group—Fmoc(TFMD)Phe—was incorporated into the sequence (Aβ*_16–22). Electrospray ionization ion-mobility spectrometry mass-spectrometry (ESI-IMS-MS) analysis of the PIC products confirmed that Aβ*_16–22 forms assemblies with the monomers arranged as anti-parallel, in-register β-strands at all time points during the aggregation assay. The assembly process was also monitored separately using fluorescence quenching to profile the fibril assembly reaction. The molecular picture resulting from discontinuous molecule dynamics simulations showed that Aβ_16-22 assembles through a single-step nucleation into a β-sheet fibril in agreement with these experimental observations. This study provides detailed structural insights into the Aβ_16-22 self-assembly processes, paving the way to explore the self-assembly mechanism of larger, more complex peptides, including those whose aggregation is responsible for human disease.     

Extrusion of highly filled flexible polymer sheet

Highly-filled polymer systems include color masterbatches, feedstocks for powder injection molding, and rigid sheets with high levels of flame retardants, but they have not been explored for flexible sheet. This work investigated the (a) selecting a polymer matrix with enough melt strength and flexibility to form a stable sheet with high filler loading, (b) the maximum achievable filler loading for the sheet, and (c) optimizing the process of extruding a highly-filled flexible polymer system. Extrusion grade low-density polyethylene (LDPE) provided sufficient flexibility and permitted a maximum filler loading of 36 vol% (~78 wt%). Good dispersion of the nanoparticle filler, however, required two passes through multiple screw extruders and a small reduction in the viscosity of the LDPE. Sheet with thickness of 415 μm, surface roughness of <1 μm, and sufficient flexibility was extruded continuously at a rate of 10 m/min., but it required a more traditional coat hanger manifold to prevent filler hang up in the sheet die. The filler particles were distributed uniformly through the core and skin of the sheet, giving the sheet good mechanical properties.     

Ubiquitin C-Terminal Hydrolase L1: Biochemical and Cellular Characterization of a Covalent Cyanopyrrolidine-Based Inhibitor

The deubiquitinase (DUB) ubiquitin C-terminal hydrolase L1 (UCHL1) is expressed primarily in the central nervous system under normal physiological conditions. However, UCHL1 is overexpressed in various aggressive forms of cancer with strong evidence supporting UCHL1 as an oncogene in lung, glioma, and blood cancers. In particular, the level of UCHL1 expression in these cancers correlates with increased invasiveness and metastatic behavior, as well as poor patient prognosis. Although UCHL1 is considered an oncogene with potential as a therapeutic target, there remains a significant lack of useful small-molecule probes to pharmacologically validate in vivo targeting of the enzyme. Herein, we describe the characterization of a new covalent cyanopyrrolidine-based UCHL1 inhibitory scaffold in biochemical and cellular studies to better understand the utility of this inhibitor in elucidating the role of UCHL1 in cancer biology.     

Impact of pre-treatment (soaking or germination) on nutrient and anti-nutrient contents,cooking time and acceptability of cooked red dry bean (Phaseolus vulgaris L.) and chickpea (Cicer arietinum L.) grown in Ethiopia

Pulses are processed in diverse ways prior to consumption. Soaking and germination are among the most common traditional, household-level food processing strategies. This study was carried out to determine the effects of soaking, germination, cooking and their combinations on the contents of selected nutrients and anti-nutrients of red dry bean and chickpea. In addition, the effects of pre-treatment on cooking time and the acceptability of dishes prepared from red dry bean and chickpea were determined. The nutrient compositions (zinc, iron and calcium) of most soaked-cooked and germinated-cooked red dry bean and chickpea samples were not significantly different than those of respective controls. However, soaking and germination pre-treatments significantly lowered the phytate and tannin contents of the red dry bean and chickpea samples, with a few exceptions, and overall, polyphenol contents were lower after soaking-cooking than after germination-cooking. Most scores for sensory attributes of bean-based and chickpea-based dishes prepared from soaked or germinated samples were not significantly different than those of the controls. For most red dry bean and chickpea samples, longer germination times yielded superior results in terms of reductions in cooking time, tannin content, and phytate:zinc and phytate:iron molar ratio.     

Collaborative analytics-supported reflective Assessment for Scaffolding Pre-service Teachers’ collaborative Inquiry and Knowledge Building

International Journal of Computer-Supported Collaborative Learning - Helping pre-service teachers (PSTs) develop competencies in collaborative inquiry and knowledge building is crucial, but this...     

Addition of carbon black NIR absorber to galvanised steel primer systems: Influence on NIR cure of polyester melamine topcoats and corrosion protection characteristics

Near infrared (NIR) is becoming a popular option for rapid cure of coatings in the coil coating industry particularly where fast line speeds are required. The technology has the potential to reduce the cure time of a 20 μm polyester coating on a galvanised steel substrate from around 30 s via conventional heating methods down to <10 s under the lamps. Previous work suggested that the ideal situation in this case is to have a topcoat which is slightly transparent to NIR and an absorbing substrate to heat the coating from the substrate outwards in a two stage process which separates solvent removal from cross linking and film formation. This can be taken further by tinting the primer layer with a pigment that absorbs in the NIR region. In this study spectroscopy was used to show that a coated steel system could appear white in the visible region because of the reflectance of TiO₂ but the NIR absorption could be altered by adding absorbing pigments such as carbon black. Lamp settings could be reduced by 20% to achieve equivalent cure with tinted primer systems. The potential degradation in corrosion protection afforded by carbon-black containing pigments at various loadings was assessed for model organic coatings applied to galvanised steel specimens. In situ scanning Kelvin probe studies showed that rates of corrosion-driven coating delamination by cathodic disbondment remained unchanged by pigment loadings of up to 3.5 wt%.     

A Meta-frontier method of decomposing long-term construction productivity components and technological gaps at the firm level: evidence from Malaysia

There is a need for a better approach to measure construction productivity rigorously, multilaterally, longitudinally and decomposed into its components. In response, this paper offers a robust approach to analysing construction productivity at the firm level that has been lacking in the industry to date, by measuring the Total Factor Productivity (TFP) of 37 public-listed Malaysian construction firms over 14 years (2003–2016), based on the Färe-Primont index. In comparing different groups of building, civil and specialist construction firms, this is the first application of a meta-frontier framework to capture the technological gaps involved. Based on the construction firms’ financial data, it is found that TFP improvement generally occurs due to Technical Efficiency (TE) and Scale-Mix Efficiency (SME) (largely scope economies), and significant technological gaps exist among different groups. Moreover, the industry suffered a decline in technologically related production environment over the period – prompting the conclusion that long-term policy engagement should focus on technological improvements. The paper provides a robust approach to analysing construction productivity at the firm level that also can be used for accessing productivity components and technological gaps in construction and other industries.     

Cellular Scent of Influenza Virus Infection.

Volatile organic compounds (VOCs) emanating from humans have the potential to revolutionize non‐invasive diagnostics. Yet, little is known about how these compounds are generated by complex biological systems, and even less is known about how these compounds are reflective of a particular physiological state. In this proof‐of‐concept study, we examined VOCs produced directly at the cellular level from B lymphoblastoid cells upon infection with three live influenza virus subtypes: H9N2 (avian), H6N2 (avian), and H1N1 (human). Using a single cell line helped to alleviate some of the complexity and variability when studying VOC production by an entire organism, and it allowed us to discern marked differences in VOC production upon infection of the cells. The patterns of VOCs produced in response to infection were unique for each virus subtype, while several other non‐specific VOCs were produced after infections with all three strains. Also, there was a specific time course of VOC release post infection. Among emitted VOCs, production of esters and other oxygenated compounds was particularly notable, and these may be attributed to increased oxidative stress resulting from infection. Elucidating VOC signatures that result from the host cells response to infection may yield an avenue for non‐invasive diagnostics and therapy of influenza and other viral infections.