Lipid-modulated assembly of magnetized iron-filled carbon nanotubes in millimeter-scale structures

Biomolecule-functionalized carbon nanotubes (CNTs) combine the molecular recognition properties of biomaterials with the electrical properties of nanoscale solid state transducers. Application of this hybrid material in bioelectronic devices requires the development of methods for the reproducible self-assembly of CNTs into higher-order structures in an aqueous environment. To this end, we have studied pattern formation of lipid-coated Fe-filled CNTs, with lengths in the 1–5 µm range, by controlled evaporation of aqueous CNT-lipid suspensions. Novel diffusion limited aggregation structures composed of end-to-end oriented nanotubes were observed by optical and atomic force microscopy. Significantly, the lateral dimension of assemblies of magnetized Fe-filled CNTs was in the millimeter range. Control experiments in the absence of lipids and without magnetization indicated that the formation of these long linear nanotube patterns is driven by a subtle interplay between radial flow forces in the evaporating droplet, lipid-modulated van der Waals forces, and magnetic dipole–dipole interactions. Keywords

A low-noise transimpedance amplifier for BLM-based ion channel recording

High-throughput screening (HTS) using ion channel recording is a powerful drug discovery technique in pharmacology. Ion channel recording with planar bilayer lipid membranes (BLM) is scalable and has very high sensitivity. A HTS system based on BLM ion channel recording faces three main challenges: (i) design of scalable microfluidic devices; (ii) design of compact ultra-low-noise transimpedance amplifiers able to detect currents in the pA range with bandwidth >10 kHz; (iii) design of compact, robust and scalable systems that integrate these two elements. This paper presents a low-noise transimpedance amplifier with integrated A/D conversion realized in CMOS 0.35 µm technology. The CMOS amplifier acquires currents in the range ±200 pA and ±20 nA, with 100 kHz bandwidth while dissipating 41 mW. An integrated digital offset compensation loop balances any voltage offsets from Ag/AgCl electrodes. The measured open-input input-referred noise current is as low as 4 fA/Root Hz at ±200 pA range. The current amplifier is embedded in an integrated platform, together with a microfluidic device, for current recording from ion channels. Gramicidin-A, alpha-haemolysin and KcsA potassium channels have been used to prove both the platform and the current-to-digital converter

Multiple stakeholders’ perspectives of marine social ecological systems, a case study on the Barents Sea

The Barents Sea ecosystem components and services are under pressure from climate change and other anthropogenic impacts. Following an Ecosystem-based management approach, multiple simultaneous pressures are addressed by using integrative strategies, but regular prioritization of key issues is needed. Identification of such priorities is typically done in a ‘scoping’ phase, where the characterization of the social-ecological system is defined and discussed. We performed a scoping exercise using an open and flexible multi-stakeholder approach to build conceptual models of the Barents Sea social-ecological system. After standardizing vocabulary, a com plex hierarchical model structure containing 155 elements was condensed to a simpler model structure con taining a maximum of 36 elements. To capture a common understanding across stakeholder groups, inputs from the individual group models were compiled into a collective model. Stakeholders’ representation of the Barents Sea social-ecological system is complex and often group specific, emphasizing the need to include social scientific methods to ensure the identification and inclusion of key stakeholders in the process. Any summary or simpli fication of the stakeholders’ representation neglects important information. Some commonalities are highlighted in the collective model, and additional information from the hierarchical model is provided by multicriteria analysis. The collective conceptual stakeholder model provides input to an integrated overview and strengthens prioritization in Ecosystem-based management by supporting the development of qualitative network models. Such models allow for exploration of perturbations and can inform cross-sectoral management trade-offs and prioritiespublishedVersio

Transmembrane helix dynamics of bacterial chemoreceptors supports a piston model of signalling.

Transmembrane α-helices play a key role in many receptors, transmitting a signal from one side to the other of the lipid bilayer membrane. Bacterial chemoreceptors are one of the best studied such systems, with a wealth of biophysical and mutational data indicating a key role for the TM2 helix in signalling. In particular, aromatic (Trp and Tyr) and basic (Arg) residues help to lock α-helices into a membrane. Mutants in TM2 of E. coli Tar and related chemoreceptors involving these residues implicate changes in helix location and/or orientation in signalling. We have investigated the detailed structural basis of this via high throughput coarse-grained molecular dynamics (CG-MD) of Tar TM2 and its mutants in lipid bilayers. We focus on the position (shift) and orientation (tilt, rotation) of TM2 relative to the bilayer and how these are perturbed in mutants relative to the wildtype. The simulations reveal a clear correlation between small (ca. 1.5 Å) shift in position of TM2 along the bilayer normal and downstream changes in signalling activity. Weaker correlations are seen with helix tilt, and little/none between signalling and helix twist. This analysis of relatively subtle changes was only possible because the high throughput simulation method allowed us to run large (n = 100) ensembles for substantial numbers of different helix sequences, amounting to ca. 2000 simulations in total. Overall, this analysis supports a swinging-piston model of transmembrane signalling by Tar and related chemoreceptors

Characterization of the Prokaryotic Sodium Channel NavSp Pore with a Microfluidic Bilayer Platform

This paper describes the use of a newly-developed micro-chip bilayer platform to examine the electrophysiological properties of the prokaryotic voltage-gated sodium channel pore (NavSp) from Silicibacter pomeroyi. The platform allows up to 6 bilayers to be analysed simultaneously. Proteoliposomes were incorporated into suspended lipid bilayers formed within the microfluidic bilayer chips. The chips provide access to bilayers from either side, enabling the fast and controlled titration of compounds. Dose-dependent modulation of the opening probability by the channel blocking drug nifedipine was measured and its IC50 determined

Operational continental-scale land cover mapping of Australia using the Open Data Cube

To comprehensively support national and international initiatives for sustainable development, land cover products need to be reliably and routinely generated within operational frameworks. Coupled with consistent semantics and taxonomies, ensuring confidence in mapping land cover for multiple time periods, facilitates informed decision-making at scales appropriate to multiple policy domains. The United Nations Food and Agriculture Organisation (FAO) Land Cover Classification System (LCCS) provides a taxonomy that comparable at different scales, level of detail and geographic location. The Open Data Cube (ODC) initiative offers a framework for operational continental scale land cover mapping using analysis-ready Earth Observation data. This study utilised the FAO LCCS framework and the Landsat sensor data through Digital Earth Australia (DEA; Australia’s ODC instance) to generate consistent and continent-wide land cover mapping (DEA Land Cover) of the Australian continent. DEA Land Cover provides annual maps from 1988 to 2020 at 25 m resolution. Output maps were validated with ∼12,000 independent validation points, giving an overall map accuracy of 80%. DEA Land Cover provides Australia with a nationally consistent picture of land cover, with an open-source software package using readily available global coverage data and demonstrates a pathway of adoption for national implementations across the worl

Orientation and dynamics of transmembrane peptides: the power of simple models

In this review we discuss recent insights obtained from well-characterized model systems into the factors that determine the orientation and tilt angles of transmembrane peptides in lipid bilayers. We will compare tilt angles of synthetic peptides with those of natural peptides and proteins, and we will discuss how tilt can be modulated by hydrophobic mismatch between the thickness of the bilayer and the length of the membrane spanning part of the peptide or protein. In particular, we will focus on results obtained on tryptophan-flanked model peptides (WALP peptides) as a case study to illustrate possible consequences of hydrophobic mismatch in molecular detail and to highlight the importance of peptide dynamics for the experimental determination of tilt angles. We will conclude with discussing some future prospects and challenges concerning the use of simple peptide/lipid model systems as a tool to understand membrane structure and function

Desmoglein 2 is a substrate of kallikrein 7 in pancreatic cancer

<p>Abstract</p> <p>Background</p> <p>In a previous report we have demonstrated that the chymotryptic-like serine protease kallikrein 7 (<it>KLK7</it>/hK7) is overexpressed in pancreatic cancer. In normal skin, hK7 is thought to participate in skin desquamation by contributing in the degradation of desmosomal components, such as desmogleins. Thus, the ability of hK7 to degrade desmogleins was assessed and the effect of hK7 expression on desmoglein 2 was examined in cultured pancreatic cancer cells.</p> <p>Methods</p> <p>The expression of Dsg1, Dsg2, and Dsg3 in pancreatic tissues was examined by immunohistochemistry and their expression in two pancreatic cancer cell lines, BxPC-3 and Panc-1, was determined by western blot analysis. The ability of hK7 to degrade Dsg1 and Dsg2 was investigated using <it>in vitro </it>degradation assays. BxPC-3 cells stably transfected to overexpress hK7 were used to examine the effect of hK7 on cell-surface resident Dsg2.</p> <p>Results</p> <p>The levels of immunoreactive Dsg1 and Dsg2 were reduced in pancreatic adenocarcinomas compared with both normal pancreatic and chronic pancreatitis tissues. Among the desmosomal proteins examined, Dsg2 exhibited robust expression on the surface of BxPC-3 cells. When hK7 was overexpressed in this cell line, there was a significant increase in the amount of soluble Dsg2 released into the culture medium compared with vector-transfected control cells.</p> <p>Conclusion</p> <p>A reduction in the amount of the cell adhesion components Dsg1 and Dsg2 in pancreatic tumors suggests that loss of these desmosomal proteins may play a role in pancreatic cancer invasion. Using <it>in vitro </it>degradation assays, both Dsg1 and Dsg2 could be readily proteolyzed by hK7, which is overexpressed in pancreatic adenocarcinomas. The enforced expression of hK7 in BxPC-3 cells that express significant amounts of Dsg2 resulted in a marked increase in the shedding of soluble Dsg2, which is consistent with the notion that aberrant expression of hK7 in pancreatic tumors may result in diminished cell-cell adhesion and facilitate tumor cell invasion.</p