Controlled delivery of membrane proteins to artificial lipid bilayers by nystatin-ergosterol modulated vesicle fusion

The study of ion channels and other membrane proteins and their potential use as biosensors and drug screening targets require their reconstitution in an artificial membrane. These applications would greatly benefit from microfabricated devices in which stable artificial lipid bilayers can be rapidly and reliably formed. However, the amount of protein delivered to the bilayer must be carefully controlled. A vesicle fusion technique is investigated where composite ion channels of the polyene antibiotic nystatin and the sterol ergosterol are employed to render protein-carrying vesicles fusogenic After fusion with an ergosterol-free artificial bilayer the nystatin-ergosterol channels do not dissociate immediately and thus cause a transient current signal that marks the vesicle fusion event. Experimental pitfalls of this method were identified, the influence of the nystatin and ergosterol concentration on the fusion rate and the shape of the fusion event marker was explored, and the number of different lipid was reduced. Under these conditions, the B-amyloid peptide could be delivered in a controlled manner to a standard planar bilayer. Additionally, the electrical recordings were obtained of vesicles fusing with a planar lipid bilayer in a microfabricated device, demonstrating the suitability of nystatin-ergosterol modulated vesicle fusion for protein delivery within microsystems

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 game-theoretic model of interspecific brood parasitism with sequential decisions

The interaction between hosts and parasites in bird populations has been studied extensively. This paper uses game-theoretic methods to model this interaction. This has been done in previous papers but has not been studied taking into account the detailed sequential nature of this game. We introduce a model allowing the host and parasite to make a number of decisions which will depend on various natural factors. The sequence of events begins with the host forming a nest and laying a number of eggs, followed by the possibility that a parasite bird will arrive at the nest; if it does it can choose to destroy some of the host eggs and lay one of its own. A sequence of events follows, which is broken down into two key stages; firstly the interaction between the host and the parasite adult, and secondly that between the host and the parasite chick. The final decision involves the host choosing whether to raise or abandon the chicks that are in the nest. There are certain natural parameters and probabilities which are central to these various decisions; in particular the host is generally uncertain whether parasitism has taken place, but can assess the likelihood of parasitism based upon certain cues (e.g. how many eggs remain in its nest). We then use this methodology to model two real-world interactions, that of the Reed Warbler with the Common Cuckoo and also the Yellow Warbler with the Brown-headed Cowbird. These parasites have different methods in the way they parasitize the nests of their hosts, and the hosts can in turn have different reactions to these parasites. Our model predictions generally match the real results well, and the model also makes predictions of the effect of changes in various key parameters on the type of parasitic interactions that should occur

Circular material flow in the intensive care unit-environmental effects and identification of hotspots

PurposeThe healthcare sector is responsible for 6–7% of CO2 emissions. The intensive care unit (ICU) contributes to these CO2 emissions and a shift from a linear system to a circular system is needed. The aim of our research was to perform a material flow analysis (MFA) in an academic ICU. Secondary aims were to obtain information and numbers on mass, carbon footprint, agricultural land occupation and water usage and to determine so-called “environmental hotspots” in the ICU.MethodsA material flow analysis was performed over the year 2019, followed by an environmental footprint analysis of materials and environmental hotspot identification.Results2839 patients were admitted to our ICU in 2019. The average length of stay was 4.6 days. Our MFA showed a material mass inflow of 247,000 kg in 2019 for intensive care, of which 50,000 kg is incinerated as (hazardous) hospital waste. The environmental impact per patient resulted in 17 kg of mass, 12 kg CO2 eq, 300 L of water usage and 4 m2 of agricultural land occupation per day. Five hotspots were identified: non-sterile gloves, isolation gowns, bed liners, surgical masks and syringes (including packaging).ConclusionThis is the first material flow analysis that identified environmental risks and its magnitude in the intensive care unit

Living Earth:Implementing national standardised land cover classification systems for Earth Observation in support of sustainable development

Earth Observation (EO) has been recognised as a key data source for supporting the United Nations Sustainable Development Goals (SDGs). Advances in data availability and analytical capabilities have provided a wide range of users access to global coverage analysis-ready data (ARD). However, ARD does not provide the information required by national agencies tasked with coordinating the implementation of SDGs. Reliable, standardised, scalable mapping of land cover and its change over time and space facilitates informed decision making, providing cohesive methods for target setting and reporting of SDGs. The aim of this study was to implement a global framework for classifying land cover. The Food and Agriculture Organisation’s Land Cover Classification System (FAO LCCS) provides a global land cover taxonomy suitable to comprehensively support SDG target setting and reporting. We present a fully implemented FAO LCCS optimised for EO data; Living Earth, an open-source software package that can be readily applied using existing national EO infrastructure and satellite data. We resolve several semantic challenges of LCCS for consistent EO implementation, including modifications to environmental descriptors, inter-dependency within the modular-hierarchical framework, and increased flexibility associated with limited data availability. To ensure easy adoption of Living Earth for SDG reporting, we identified key environmental descriptors to provide resource allocation recommendations for generating routinely retrieved input parameters. Living Earth provides an optimal platform for global adoption of EO4SDGs ensuring a transparent methodology that allows monitoring to be standardised for all countrie

Effect of Phosphate Buffered Saline Solutions on Top-Down Fabricated ZnO Nanowire Field Effect Transistor

ZnO based nanowire FETs have been fabricated by implementing a top-down approach, which uses optical photolithography, atomic layer deposition (ALD) of ZnO thin film, and anisotropic plasma etching. The effects of Phosphate Buffered Saline (PBS) solution on the surface of ZnO nanowire were investigated by measuring the FET characteristics at different PBS dilutions. The drain current, ION, exhibited an increase of 39 times in the highest PBS solution concentration compared to measurement in air. From the measured transfer characteristics and output characteristics in various PBS dilutions, the device was found to maintain n-type behaviour. These results indicate that the device can be effectively used for biomolecules sensing

CCN3 controls 3D spatial localization of melanocytes in the human skin through DDR1

Melanocytes reside within the basal layer of the human epidermis, where they attach to the basement membrane and replicate at a rate proportionate to that of keratinocytes, maintaining a lifelong stable ratio. In this study, we report that coculturing melanocytes with keratinocytes up-regulated CCN3, a matricellular protein that we subsequently found to be critical for the spatial localization of melanocytes to the basement membrane. CCN3 knockdown cells were dissociated either upward to the suprabasal layers of the epidermis or downward into the dermis. The overexpression of CCN3 increased adhesion to collagen type IV, the major component of the basement membrane. As the receptor responsible for CCN3-mediated melanocyte localization, we identified discoidin domain receptor 1 (DDR1), a receptor tyrosine kinase that acts as a collagen IV adhesion receptor. DDR1 knockdown decreased melanocyte adhesion to collagen IV and shifted melanocyte localization in a manner similar to CCN3 knockdown. These results demonstrate an intricate and necessary communication between keratinocytes and melanocytes in maintaining normal epidermal homeostasis

CD40 signal rewires fatty acid and glutamine metabolism for stimulating macrophage anti-tumorigenic functions.

Exposure of lipopolysaccharide triggers macrophage pro-inflammatory polarization accompanied by metabolic reprogramming, characterized by elevated aerobic glycolysis and a broken tricarboxylic acid cycle. However, in contrast to lipopolysaccharide, CD40 signal is able to drive pro-inflammatory and anti-tumorigenic polarization by some yet undefined metabolic programming. Here we show that CD40 activation triggers fatty acid oxidation (FAO) and glutamine metabolism to promote ATP citrate lyase-dependent epigenetic reprogramming of pro-inflammatory genes and anti-tumorigenic phenotypes in macrophages. Mechanistically, glutamine usage reinforces FAO-induced pro-inflammatory and anti-tumorigenic activation by fine-tuning the NAD <sup>+</sup> /NADH ratio via glutamine-to-lactate conversion. Genetic ablation of important metabolic enzymes involved in CD40-mediated metabolic reprogramming abolishes agonistic anti-CD40-induced antitumor responses and reeducation of tumor-associated macrophages. Together these data show that metabolic reprogramming, which includes FAO and glutamine metabolism, controls the activation of pro-inflammatory and anti-tumorigenic polarization, and highlight a therapeutic potential of metabolic preconditioning of tumor-associated macrophages before agonistic anti-CD40 treatments

The application of predictive modelling for determining bio-environmental factors affecting the distribution of blackflies (Diptera: Simuliidae) in the Gilgel Gibe watershed in Southwest Ethiopia

Blackflies are important macroinvertebrate groups from a public health as well as ecological point of view. Determining the biological and environmental factors favouring or inhibiting the existence of blackflies could facilitate biomonitoring of rivers as well as control of disease vectors. The combined use of different predictive modelling techniques is known to improve identification of presence/absence and abundance of taxa in a given habitat. This approach enables better identification of the suitable habitat conditions or environmental constraints of a given taxon. Simuliidae larvae are important biological indicators as they are abundant in tropical aquatic ecosystems. Some of the blackfly groups are also important disease vectors in poor tropical countries. Our investigations aim to establish a combination of models able to identify the environmental factors and macroinvertebrate organisms that are favourable or inhibiting blackfly larvae existence in aquatic ecosystems. The models developed using macroinvertebrate predictors showed better performance than those based on environmental predictors. The identified environmental and macroinvertebrate parameters can be used to determine the distribution of blackflies, which in turn can help control river blindness in endemic tropical places. Through a combination of modelling techniques, a reliable method has been developed that explains environmental and biological relationships with the target organism, and, thus, can serve as a decision support tool for ecological management strategies

Recognition Pliability Is Coupled to Structural Heterogeneity: A Calmodulin Intrinsically Disordered Binding Region Complex

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