1. |
- Fisher, Matthew C., et al.
(author)
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Development and worldwide use of non-lethal, and minimal population-level impact, protocols for the isolation of amphibian chytrid fungi
- 2018
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In: Scientific Reports. - : NATURE PUBLISHING GROUP. - 2045-2322. ; 8
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Journal article (peer-reviewed)abstract
- Parasitic chytrid fungi have emerged as a significant threat to amphibian species worldwide, necessitating the development of techniques to isolate these pathogens into culture for research purposes. However, early methods of isolating chytrids from their hosts relied on killing amphibians. We modified a pre-existing protocol for isolating chytrids from infected animals to use toe clips and biopsies from toe webbing rather than euthanizing hosts, and distributed the protocol to researchers as part of the BiodivERsA project RACE; here called the RML protocol. In tandem, we developed a lethal procedure for isolating chytrids from tadpole mouthparts. Reviewing a database of use a decade after their inception, we find that these methods have been applied across 5 continents, 23 countries and in 62 amphibian species. Isolation of chytrids by the non-lethal RML protocol occured in 18% of attempts with 207 fungal isolates and three species of chytrid being recovered. Isolation of chytrids from tadpoles occured in 43% of attempts with 334 fungal isolates of one species (Batrachochytrium dendrobatidis) being recovered. Together, these methods have resulted in a significant reduction and refinement of our use of threatened amphibian species and have improved our ability to work with this group of emerging pathogens.
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2. |
- Roberts, O. W., et al.
(author)
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Estimation of the Electron Density From Spacecraft Potential During High-Frequency Electric Field Fluctuations
- 2020
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In: Journal of Geophysical Research - Space Physics. - : AMER GEOPHYSICAL UNION. - 2169-9380 .- 2169-9402. ; 125:9
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Journal article (peer-reviewed)abstract
- Spacecraft potential has often been used to infer electron density with much higher time resolution than is typically possible with plasma instruments. However, recently, two studies by Torkar et al. (2017, https://doi.org/10.1002/2017JA024724) and Graham, Vaivads, Khotyaintsev, Eriksson, et al. (2018, https://doi.org/10.1029/2018JA025874) have shown that external electric fields can also have an effect on the spacecraft potential by enhancing photoelectron escape from the surface. Consequently, should the electron density derived from the spacecraft potential be used during an event with a large electric field, the estimation would be contaminated and the user would see the effects of the electric field rather than density perturbations. The goal of this paper is to propose a method to remove the electric field effects to allow the density derived from spacecraft potential to be used even during large-amplitude wave events such as Langmuir waves or upper hybrid waves. Plain Language Summary Spacecraft in a plasma become charged due to a number of processes. Often the two most important processes in determining the charge are due to the ambient plasma and the photoelectron emission from the surface of a sunlit spacecraft. The potential itself is a function of the electron density, and consequently, the potential data can be used to infer the electron density if the photoelectron emission can be modeled. However, in the presence of large electric fields, the photoelectron emission can change with the electric field. This means that rather than see fluctuations of density in the spacecraft potential, the effect of the electric field is seen. Here a method is presented to remove the electric field effect on the spacecraft potential such that the density can be estimated even when there are strong electric fields present.
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3. |
- Bally, Marta, 1981, et al.
(author)
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Fluorescent vesicles for signal amplification in reverse phase protein microarray assays
- 2011
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In: Analytical Biochemistry. - 0003-2697 .- 1096-0309. ; 416:2, s. 145-151
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Journal article (peer-reviewed)abstract
- Developments in microarray technology promise to lead to great advancements in the biomedical and biological field. However, implementation of these analytical tools often relies on signal amplification strategies that are essential to reach the sensitivity levels required for a variety of biological applications. This is true especially for reverse phase arrays where a complex biological sample is directly immobilized on the chip. We present a simple and generic method for signal amplification based on the use of antibody-tagged fluorescent vesicles as labels for signal generation. To assess the gain in assay sensitivity, we performed a model assay for the detection of rabbit immunoglobulin G (IgG) and compared the limit of detection (LOD) of the vesicle assay with the LOD of a conventional assay performed with fluorescent reporter molecules. We evaluated the improvements for two fluorescence-based transduction setups: a high-sensitivity microarray reader (ZeptoREADER) and a conventional confocal scanner. In all cases, our strategy led to an increase in sensitivity. However, gain in sensitivity widely depended on the type of illumination; whereas an approximately 2-fold increase in sensitivity was observed for readout based on evanescent field illumination, the contribution was as high as more than 200-fold for confocal scanning. (C) 2011 Elsevier Inc. All rights reserved.
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4. |
- Bally, Marta, 1981, et al.
(author)
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Liposome and lipid bilayer arrays towards biosensing applications
- 2010
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In: Small. - : Wiley. - 1613-6810 .- 1613-6829. ; 6:22, s. 2481-2497
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Journal article (peer-reviewed)abstract
- Sensitive and selective biosensors for high-throughput screening are having an increasing impact in modern medical care. The establishment of robust protein biosensing platforms however remains challenging, especially when membrane proteins are involved. Although this type of proteins is of enormous relevance since they are considered in >60% of the pharmaceutical drug targets, their fragile nature (i.e., the requirement to preserve their natural lipid environment to avoid denaturation and loss of function) puts strong additional prerequisites onto a successful biochip. In this review, the leading approaches to create lipid membrane-based arrays towards the creation of membrane protein biosensing platforms are described. Liposomes assembled in micro- and nanoarrays and the successful set-ups containing functional membrane proteins, as well as the use of liposomes in networks, are discussed in the first part. Then, the complementary approaches to create cell-mimicking supported membrane patches on a substrate in an array format will be addressed. Finally, the progress in assembling free-standing (functional) lipid bilayers over nanopore arrays for ion channel sensing will be reported. This review illustrates the rapid pace by which advances are being made towards the creation of a heterogeneous biochip for the high-throughput screening of membrane proteins for diagnostics, drug screening, or drug discovery purposes.
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5. |
- Boros, Emil, et al.
(author)
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Macrophytes and groundwater drive extremely high organic carbon concentration of soda pans
- 2020
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In: Freshwater Biology. - : Wiley. - 0046-5070 .- 1365-2427. ; 65:9, s. 1555-1568
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Journal article (peer-reviewed)abstract
- Endorheic soda pans are among the highest dissolved organic carbon (DOC) content aquatic systems on the planet with concentrations up to 1 g/L. Considering the importance of inland waters in the global carbon cycle, understanding the drivers of such outstanding organic carbon pools is eminent. The soda pans of the Carpathian Basin present a wide variability of biotic and abiotic characteristics that provide an adequate system to assess the determinants of extreme high DOC concentrations. Here, we demonstrate through a multi‐site comparison, a multi‐year seasonal monitoring, and a laboratory experiment that the dissolved organic matter content of the highest DOC concentration soda pans is primarily of groundwater and emergent macrophyte origin.More precisely, the multi‐site comparison of 14 soda pans revealed that variation of coloured dissolved organic matter (CDOM) content of the surface water of soda pans is partially explained by the CDOM content (22% of variation) of local groundwater, indicating the significant role of allochthonous terrestrial DOC sources. Further 23% of CDOM variation could be accounted for by Bolboschoenus maritimus species‐specific emergent macrophyte cover, while the contribution of Phragmites australis cover was only minor.In line with the results of the multi‐site comparison, our decomposition experiment demonstrated that both B. maritimus and P. australis have the potential to release substantial amount of organic matter into soda pans. However, the organic matter release of B. maritimus leads to twice as high DOC and 3.5‐times higher CDOM concentrations than P. australis. Considering previous organic matter release studies, we concluded that P. australis is a relatively low organic matter releaser emergent macrophyte, and therefore the species composition of emergent macrophytes has to be carefully considered in autochthonous plant‐derived DOM estimations.Finally, the multi‐year seasonal monitoring of two distinctive soda pans showed that the high organic matter concentrations depend not only on their intrinsic characteristics but also on interannual variability. More precisely, we demonstrated that the highest CDOM and DOC concentrations that occurred in a coloured (i.e. brown, low total suspended solids) soda pan with extensive (95%) macrophyte cover dominated by B. maritimus were measured in a period characterised by high pH and low water levels, which were presumably the consequence of increased evaporation due to decreased precipitation and above average temperature.Our results indicate that considering climate change trends common for most endorheic regions (i.e. increased temperature and modified precipitation regimes), extremely high organic matter concentrations might become more frequent in the near future in local water bodies, particularly in those highly influenced by groundwater inflow. Furthermore, soda pans with vast specific macrophyte cover and substantial groundwater inflow might become organic carbon processing hotspots.
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6. |
- Dahlin, Andreas, 1980, et al.
(author)
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Electrochemical plasmonic sensors
- 2012
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In: Analytical and Bioanalytical Chemistry. - : Springer Science and Business Media LLC. - 1618-2642 .- 1618-2650. ; 402:5, s. 1773-1784
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Research review (peer-reviewed)abstract
- The enormous progress of nanotechnology during the last decade has made it possible to fabricate a great variety of nanostructures. On the nanoscale, metals exhibit special electrical and optical properties, which can be utilized for novel applications. In particular, plasmonic sensors including both the established technique of surface plasmon resonance and more recent nanoplasmonic sensors, have recently attracted much attention. However, some of the simplest and most successful sensors, such as the glucose biosensor, are based on electrical readout. In this review we describe the implementation of electrochemistry with plasmonic nanostructures for combined electrical and optical signal transduction. We highlight results from different types of metallic nanostructures such as nanoparticles, nanowires, nanoholes or simply films of nanoscale thickness. We briefly give an overview of their optical properties and discuss implementation of electrochemical methods. In particular, we review studies on how electrochemical potentials influence the plasmon resonances in different nanostructures, as this type of fundamental understanding is necessary for successful combination of the methods. Although several combined platforms exist, many are not yet in use as sensors partly because of the complicated effects from electrochemical potentials on plasmon resonances. Yet, there are clearly promising aspects of these sensor combinations and we conclude this review by discussing the advantages of synchronized electrical and optical readout, illustrating the versatility of these technologies.
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7. |
- Dahlin, Andreas, 1980, et al.
(author)
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Nanoplasmonic sensing of metal-halide complex formation and the electric double layer capacitor
- 2012
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In: Nanoscale. - : Royal Society of Chemistry (RSC). - 2040-3372 .- 2040-3364. ; 4:7, s. 2339-2351
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Journal article (peer-reviewed)abstract
- Many nanotechnological devices are based on implementing electrochemistry with plasmonic nanostructures, but these systems are challenging to understand. We present a detailed study of the influence of electrochemical potentials on plasmon resonances, in the absence of surface coatings and redox active molecules, by synchronized voltammetry and spectroscopy. The experiments are performed on gold nanodisks and nanohole arrays in thin gold films, which are fabricated by improved methods. New insights are provided by high resolution spectroscopy and variable scan rates. Furthermore, we introduce new analytical models in order to understand the spectral changes quantitatively. In contrast to most previous literature, we find that the plasmonic signal is caused almost entirely by the formation of ionic complexes on the metal surface, most likely gold chloride in this study. The refractometric sensing effect from the ions in the electric double layer can be fully neglected, and the charging of the metal gives a surprisingly small effect for these systems. Our conclusions are consistent for both localized nanoparticle plasmons and propagating surface plasmons. We consider the results in this work especially important in the context of combined electrochemical and optical sensors.
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8. |
- de Lange, V., et al.
(author)
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Microarrays Made Easy: Biofunctionalized Hydrogel Channels for Rapid Protein Microarray Production
- 2011
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In: ACS Applied Materials & Interfaces. - : American Chemical Society (ACS). - 1944-8252 .- 1944-8244. ; 3:1, s. 50-57
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Journal article (peer-reviewed)abstract
- We present a simple, inexpensive, and sensitive technique for producing multiple copies of a hydrogel-based protein microarray. An agarose block containing 25 biofunctionalized channels is sliced perpendicularly to produce many identical biochips. Each microarray consists of 500 mu m spots, which contain protein-coated microparticles physically trapped in porous SeaPrep agarose. Proteins diffuse readily through SeaPrep agarose while the larger microparticles are immobilized in the hydrogel matrix Without major assay optimization the limit of detection is 12 pM for a sandwich assay detecting human IgG. These highly flexible, multiplexed arrays can produced rapidly without any special instrumentation and are compatible with standard fluorescence-based read-out.
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9. |
- Junesch, Juliane, 1987, et al.
(author)
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Location-specific nanoplasmonic sensing of biomolecular binding to lipid membranes with negative curvature
- 2015
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In: Nanoscale. - : Royal Society of Chemistry (RSC). - 2040-3372 .- 2040-3364. ; 7:37, s. 15080-15085
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Journal article (peer-reviewed)abstract
- The biochemical processes of cell membranes are sensitive to the geometry of the lipid bilayer. We show how plasmonic "nanowells" provide label-free real-time analysis of molecules on membranes with detection of preferential binding at negative curvature. It is demonstrated that norovirus accumulate in invaginations due to multivalent interactions with glycosphingolipids.
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10. |
- MacKenzie, R., et al.
(author)
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Simultaneous electrical and plasmonic monitoring of potential induced ion adsorption on metal nanowire arrays
- 2013
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In: Nanoscale. - : Royal Society of Chemistry (RSC). - 2040-3372 .- 2040-3364. ; 5:11, s. 4966-4975
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Journal article (peer-reviewed)abstract
- Simultaneous LSPR and electronic sensing of potential induced ion adsorption onto gold nanowire arrays is presented. The formation of a Stern layer upon applying an electrochemical potential generated a complex optical response. Simulation of a lossy atomic layer on the nanowire array using the Multiple Multipole Program (MMP) corresponded very well to the experimentally observed peak position, intensity, and radius of curvature changes. Additionally, a significant voltage-dependent change in the resistance of the gold nanowire array was observed during the controlled formation of the electrical double layer. The results demonstrated that an applied electrochemical potential induces measurable changes in the optical and electrical properties of the gold nanowire surface. This is the first demonstration of combined plasmonic and nanowire resistance-based sensing of a surface process in the literature.
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