Rapid fabrication of polymer microfluidic systems for the production of artificial lipid bilayers

A polymer microfluidic device has been fabricated using rapid prototyping techniques. The device was built up to allow the formation and subsequent investigation of artificial bilayer lipid membranes (BLMs). A simple dry film photoresist stamp was used to hot emboss microfluidic channels into PMMA films. Laser micromachining was employed to form an aperture into PMMA films. Laser micromachining was employed to form an aperture through the PMMA channels, across which the BLM was later formed. The dry film phororesist was also used as a simple etch mask for the deep etching of glass substrates in buffered HF solutions, which was used in this work for the production of glass embossing stamps. We show that bilayer films can be successfully produced across laser micromachined apertures in PMMA films

A Method to Determine the In-Air Spatial Spread of Clinical Electron Beams

We propose and analyze in detail a method to measure the in-air spatial spread parameter of clinical electron beams. Measurements are performed at the center of the beam and below the adjustable collimators sited in asymmetrical configuration in order to avoid the distortions due to the presence of the applicator. The main advantage of our procedure lies in the fact that the dose profiles are fitted by means of a function which includes, additionally to the Gaussian step usually considered, a background which takes care of the dose produced by different mechanisms that the Gaussian model does not account for. As a result, the spatial spread is obtained directly from the fitting procedure and the accuracy permits a good determination of the angular spread. The way the analysis is done is alternative to that followed by the usual methods based on the evaluation of the penumbra width. Besides, the spatial spread found shows the quadratic-cubic dependence with the distance to the source predicted by the Fermi-Eyges theory. However, the corresponding values obtained for the scattering power are differing from those quoted by ICRU nr. 35 by a factor ~2 or larger, what requires of a more detailed investigation.Comment: 11 pages, 5 Postscript figures, to be published in Medical Physic

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

Changes in vitamin biomarkers during a 2-year intervention trial involving increased fruit and vegetable consumption by free-living volunteers

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Development of a Proton Radiation Therapy Facility at IUCF

This research was sponsored by the National Science Foundation Grant NSF PHY-931478

Status of the IUCF Proton Radiation Therapy Facility

This research was sponsored by the National Science Foundation Grant NSF PHY-931478

Quantification of functionalised gold nanoparticle-targeted knockdown of gene expression in HeLa cells

Introduction: Gene therapy continues to grow as an important area of research, primarily because of its potential in the treatment of disease. One significant area where there is a need for better understanding is in improving the efficiency of oligonucleotide delivery to the cell and indeed, following delivery, the characterization of the effects on the cell. Methods: In this report, we compare different transfection reagents as delivery vehicles for gold nanoparticles functionalized with DNA oligonucleotides, and quantify their relative transfection efficiencies. The inhibitory properties of small interfering RNA (siRNA), single-stranded RNA (ssRNA) and single-stranded DNA (ssDNA) sequences targeted to human metallothionein hMT-IIa are also quantified in HeLa cells. Techniques used in this study include fluorescence and confocal microscopy, qPCR and Western analysis. Findings: We show that the use of transfection reagents does significantly increase nanoparticle transfection efficiencies. Furthermore, siRNA, ssRNA and ssDNA sequences all have comparable inhibitory properties to ssDNA sequences immobilized onto gold nanoparticles. We also show that functionalized gold nanoparticles can co-localize with autophagosomes and illustrate other factors that can affect data collection and interpretation when performing studies with functionalized nanoparticles. Conclusions: The desired outcome for biological knockdown studies is the efficient reduction of a specific target; which we demonstrate by using ssDNA inhibitory sequences targeted to human metallothionein IIa gene transcripts that result in the knockdown of both the mRNA transcript and the target protein

Understanding carbon sequestration from nature-based solutions

The International Union for the Conservation of Nature (IUCN) defines nature-based solutions (NBS) as “actions to protect, sustainably manage, and restore natural or modified ecosystems that address societal challenges effectively and adaptively, simultaneously providing human well-being and biodiversity benefits”. We assessed evidence for the greenhouse gas (GHG) mitigation potential of four nature-based solutions in Scotland (agroforestry, hedgerows, un-cultivated riparian buffer zones and the restoration of species-rich grasslands) and how these can help mitigate the impacts of climate change and reduce biodiversity loss. In addition, we provided a synthesis of the strength of evidence for including these as part of Net Zero policy objectives and carbon codes

Magnetite-doped polydimethylsiloxane (PDMS) for phosphopeptide enrichment

Reversible phosphorylation plays a key role in numerous biological processes. Mass spectrometry-based approaches are commonly used to analyze protein phosphorylation, but such analysis is challenging, largely due to the low phosphorylation stoichiometry. Hence, a number of phosphopeptide enrichment strategies have been developed, including metal oxide affinity chromatography (MOAC). Here, we describe a new material for performing MOAC that employs a magnetite-doped polydimethylsiloxane (PDMS), that is suitable for the creation of microwell array and microfluidic systems to enable low volume, high throughput analysis. Incubation time and sample loading were explored and optimized and demonstrate that the embedded magnetite is able to enrich phosphopeptides. This substrate-based approach is rapid, straightforward and suitable for simultaneously performing multiple, low volume enrichments

A simplified extractive metallurgy exercise to demonstrate selective extraction of copper

A laboratory activity is described for senior high school or 1st year undergraduate level students that illustrates key concepts linked to extractive metallurgy. This experiment demonstrates preferential binding of a methoxyphenolic oxime ligand to Cu2+ in the presence of other transition metal ions in aqueous solution. The students are tasked to investigate the importance of the spatial relationship between the oxime and phenol group of two potential ligands by performing a series of short and simple tests. The objective is achieved by identifying which methoxyphenolic oxime ligand selectively and rapidly forms an isolable precipitate with Cu2+ ions. Only one of the potential ligands exhibits a clear preference for binding to Cu2+ and this can easily be identified visually (and confirmed by UV-visible spectroscopy if desired). The experiment has successfully been employed as part of an extracurricular laboratory course involving groups of 25-30 high school students.PostprintPeer reviewe