Fluorescence probe techniques to monitor protein adsorption-induced conformation changes on biodegradable polymers

The study of protein adsorption and any associated conformational changes on interaction with biomaterials is of great importance in the area of implants and tissue constructs. This study aimed to evaluate some fluorescent techniques to probe protein conformation on a selection of biodegradable polymers currently under investigation for biomedical applications. Because of the fluorescence emanating from the polymers, the use of monitoring intrinsic protein fluorescence was precluded. A highly solvatochromic fluorescent dye, Nile red, and a well-known protein label, fluorescein isothiocyanate, were employed to study the adsorption of serum albumin to polycaprolactone and to some extent also to two starch-containing polymer blends (SPCL and SEVA-C). A variety of fluorescence techniques, steady state, time resolved, and imaging were employed. Nile red was found to leach from the protein, while fluorescein isothiocyanate proved useful in elucidating a conformational change in the protein and the observation of protein aggregates adsorbed to the polymer surface. These effects were seen by making use of the phenomenon of energy migration between the fluorescent tags to monitor interprobe distance and the use of fluorescence lifetime imaging to ascertain the surface packing of the protein on polymer

Application of fluorescence techniques to the study of protein adsorption and packing on biomaterial surfaces

[Excerpt] The ways proteins compete for the surface of biomaterials and change conformation are believed to be important for the host response to implants. It is possible to elucidate information on packing and any induced conformational change by making use of different fluorescence techniques on fluorescently labelled proteins. Employing probe-probe resonance energy transfer (RET) allows inter and intra protein interactions to be distinguished. Homo resonance energy transfer (hRET) avoids many problems with having two different probes and means that labelling and subsequent purification can be done in one step. [...]Portuguese Foundation for Science and Technology, project PROTEOLIGHT (PTDC/FIS/68517/2006) and J.B. grant SFRH/BPD/17584/2004. European Union NoE EXPERTISSUES (NMP3-CT-2004-500283) and European Union FP6 STREP project HIPPOCRATES (NMP3-CT-2003-505758).info:eu-repo/semantics/publishedVersio

The impacts of environmental warming on Odonata: a review

Climate change brings with it unprecedented rates of increase in environmental temperature, which will have major consequences for the earth's flora and fauna. The Odonata represent a taxon that has many strong links to this abiotic factor due to its tropical evolutionary history and adaptations to temperate climates. Temperature is known to affect odonate physiology including life-history traits such as developmental rate, phenology and seasonal regulation as well as immune function and the production of pigment for thermoregulation. A range of behaviours are likely to be affected which will, in turn, influence other parts of the aquatic ecosystem, primarily through trophic interactions. Temperature may influence changes in geographical distributions, through a shifting of species' fundamental niches, changes in the distribution of suitable habitat and variation in the dispersal ability of species. Finally, such a rapid change in the environment results in a strong selective pressure towards adaptation to cope and the inevitable loss of some populations and, potentially, species. Where data are lacking for odonates, studies on other invertebrate groups will be considered. Finally, directions for research are suggested, particularly laboratory studies that investigate underlying causes of climate-driven macroecological patterns

Instrumentation for fluorescence lifetime measurement using photon counting

We describe the evolution of HORIBA Jobin Yvon IBH Ltd, and its time-correlated single-photon counting (TCSPC) products, from university research beginnings through to its present place as a market leader in fluorescence lifetime spectroscopy. The company philosophy is to ensure leading-edge research capabilities continue to be incorporated into instruments in order to meet the needs of the diverse range of customer applications, which span a multitude of scientific and engineering disciplines. We illustrate some of the range of activities of a scientific instrument company in meeting this goal and highlight by way of an exemplar the performance of the versatile DeltaFlex instrument in measuring fluorescence lifetimes. This includes resolving fluorescence lifetimes down to 5 ps, as frequently observed in energy transfer, nanoparticle metrology with sub-nanometre resolution and measuring a fluorescence lifetime in as little as 60 μs for the study of transient species and kinetics

Two-photon dual imaging platform for in vivo monitoring cellular oxidative stress in liver injury

Oxidative stress reflects an imbalance between reactive oxygen species (ROS) and antioxidants, which has been reported as an early unifying event in the development and progression of various diseases and as a direct and mechanistic indicator of treatment response. However, highly reactive and short-lived nature of ROS and antioxidant limited conventional detection agents, which are influenced by many interfering factors. Here, we present a two-photon sensing platform for in vivo dual imaging of oxidative stress at the single cell-level resolution. This sensing platform consists of three probes, which combine the turn-on fluorescent transition-metal complex with different specific responsive groups for glutathione (GSH), hydrogen peroxide (H2O2) and hypochlorous acid (HOCl). By combining fluorescence intensity imaging and fluorescence lifetime imaging, these probes totally remove any possibility of crosstalk from in vivo environmental or instrumental factors, and enable accurate localization and measurement of the changes in ROS and GSH within the liver. This precedes changes in conventional biochemical and histological assessments in two distinct experimental murine models of liver injury. The ability to monitor real-time cellular oxidative stress with dual-modality imaging has significant implications for high-accurate, spatially configured and quantitative assessment of metabolic status and drug response

A global agenda for advancing freshwater biodiversity research

This manuscript is a contribution of the Alliance for Freshwater Life (www.allianceforfreshwaterlife.org). We thank Nick Bond, Lisa Bossenbroek, Lekima Copeland, Dean Jacobsen, Maria Cecilia Londo?o, David Lopez, Jaime Ricardo Garcia Marquez, Ketlhatlogile Mosepele, Nunia Thomas-Moko, Qiwei Wei and the authors of Living Waters: A Research Agenda for the Biodiversity of Inland and Coastal Waters for their contributions. We also thank Peter Thrall, Ian Harrison and two anonymous referees for their valuable comments that helped improve the manuscript. Open access funding enabled and organised by Projekt DEAL

Tracking CNS and systemic sources of oxidative stress during the course of chronic neuroinflammation

The functional dynamics and cellular sources of oxidative stress are central to understanding MS pathogenesis but remain elusive, due to the lack of appropriate detection methods. Here we employ NAD(P)H fluorescence lifetime imaging to detect functional NADPH oxidases (NOX enzymes) in vivo to identify inflammatory monocytes, activated microglia, and astrocytes expressing NOX1 as major cellular sources of oxidative stress in the central nervous system of mice affected by experimental autoimmune encephalomyelitis (EAE). This directly affects neuronal function in vivo, indicated by sustained elevated neuronal calcium. The systemic involvement of oxidative stress is mirrored by overactivation of NOX enzymes in peripheral CD11b(+) cells in later phases of both MS and EAE. This effect is antagonized by systemic intake of the NOX inhibitor and anti-oxidant epigallocatechin-3-gallate. Together, this persistent hyper-activation of oxidative enzymes suggests an "oxidative stress memory" both in the periphery and CNS compartments, in chronic neuroinflammation

Winners and losers over 35 years of dragonfly and damselfly distributional change in Germany

Aim: Recent studies suggest insect declines in parts of Europe; however, the generality of these trends across different taxa and regions remains unclear. Standardized data are not available to assess large-scale, long-term changes for most insect groups but opportunistic citizen science data are widespread for some. Here, we took advantage of citizen science data to investigate distributional changes of Odonata. Location: Germany. Methods: We compiled over 1 million occurrence records from different regional databases. We used occupancy-detection models to account for imperfect detection and estimate annual distributions for each species during 1980–2016 within 5 × 5 km quadrants. We also compiled data on species attributes that were hypothesized to affect species’ sensitivity to different drivers and related them to the changes in species’ distributions. We further developed a novel approach to cluster groups of species with similar patterns of distributional change to represent multispecies indicators. Results: More species increased (45%) than decreased (29%) or remained stable (26%) in their distribution (i.e. number of occupied quadrants). Species showing increases were generally warm-adapted species and/or running water species, while species showing decreases were cold-adapted species using standing water habitats such as bogs. Time series clustering defined five main patterns of change—each associated with a specific combination of species attributes, and confirming the key roles of species’ temperature and habitat preferences. Overall, our analysis predicted that mean quadrant-level species richness has increased over most of the time period. Main conclusions: Trends in Odonata provide mixed news—improved water quality, coupled with positive impacts of climate change, could explain the positive trends of many species. At the same time, declining species point to conservation challenges associated with habitat loss and degradation. Our study demonstrates the great value of citizen science and the work of natural history societies for assessing large-scale distributional change