Spatio-temporal variation in oxidative status regulation in a small mammal.

Life-history allocation trade-offs are dynamic over time and space according to the ecological and demographical context. Fluctuations in food availability can affect physiological trade-offs like oxidative status regulation, reflecting the balance between pro-oxidant production and antioxidant capacity. Monitoring the spatio-temporal stability of oxidative status in natural settings may help understanding its importance in ecological and evolutionary processes. However, few studies have yet conducted such procedures in wild populations. Here, we monitored individual oxidative status in a wild eastern chipmunk (Tamias striatus) population across the 2017 summer active period and over three study sites. Oxidative damage (MDA: Malondialdehyde levels) and non-enzymatic antioxidant levels (FRAP: Ferric reducing antioxidant power and HASC: Hypochlorous acid shock capacity) were quantified across time and space using assays optimized for small blood volumes. Our results showed an increase in oxidative damage mirrored by a decrease in FRAP throughout the season. We also found different antioxidant levels among our three study sites for both markers. Our results also revealed the effects of sex and body mass on oxidative status. Early in the active season, females and individuals with a greater body mass had higher oxidative damage. Males had higher HASC levels than females throughout the summer. This study shows that oxidative status regulation is a dynamic process that requires a detailed spatial and temporal monitoring to yield a complete picture of possible trade-offs between pro-oxidant production and antioxidant capacity.

Development of small blood volume assays for the measurement of oxidative stress markers in mammals.

Measuring oxidative stress has become increasingly valuable in ecological studies, especially when different markers are measured on the same individual. However, many of the current methods lack sensitivity for analysis of low blood volume samples, which represent a challenge for longitudinal field studies of small organisms. Small blood volumes can usually only be analysed by using a single assay, therefore providing limited information on individual's oxidative profile. In this study, we used blood collected from a population of wild eastern chipmunks (Tamias striatus) and modified methods presented in the literature to improve analytical selectivity and sensitivity required for small blood volumes. Specifically, we proposed a modified malondialdehyde (MDA) analysis protocol by HPLC and also optimized both the uric acid independent ferric reducing antioxidant power (FRAP) and hypochlorous acid shock capacity (HASC) assays. Development of the three modified methods was achieved with a sensitivity and repeatability that meets standards of field ecology while allowing measurement of all three assays in duplicate using less than 60 µL of plasma. Availability of these tests using small blood volumes will provide ecologists with a more comprehensive portrait of an individual's oxidative profile and a better understanding of its determinants and interactions with the environment.

Cry6Aa1, a Bacillus thuringiensis nematocidal and insecticidal toxin, forms pores in planar lipid bilayers at extremely low concentrations and without the need of proteolytic processing.

Cry6Aa1 is a Bacillus thuringiensis (Bt) toxin active against nematodes and corn rootworm insects. Its 3D molecular structure, which has been recently elucidated, is unique among those known for other Bt toxins. Typical three-domain Bt toxins permeabilize receptor-free planar lipid bilayers (PLBs) by forming pores at doses in the 1-50 µg/ml range. Solubilization and proteolytic activation are necessary steps for PLB permeabilization. In contrast to other Bt toxins, Cry6Aa1 formed pores in receptor-free bilayers at doses as low as 200 pg/ml in a wide range of pH (5.5-9.5) and without the need of protease treatment. When Cry6Aa1 was preincubated with Western corn rootworm (WCRW) midgut juice or trypsin, 100 fg/ml of the toxin was sufficient to form pores in PLBs. The overall biophysical properties of the pores were similar for all three forms of the toxin (native, midgut juice- and trypsin-treated), with conductances ranging from 28 to 689 pS, except for their ionic selectivity, which was slightly cationic for the native and midgut juice-treated Cry6Aa1, whereas dual selectivity (to cations or anions) was observed for the pores formed by the trypsin-treated toxin. Enrichment of PLBs with WCRW midgut brush-border membrane material resulted in a 2000-fold reduction of the amount of native Cry6Aa1 required to form pores and affected the biophysical properties of both the native and trypsin-treated forms of the toxin. These results indicate that, although Cry6Aa1 forms pores, the molecular determinants of its mode of action are significantly different from those reported for other Bt toxins.

Protocol for the preparation of stimuli-responsive gold nanoparticles capped with elastin-based pentapeptides.

Stimuli-responsive materials are playing an increasingly important role in a wide range of applications such as drug delivery, diagnostics, sensors, and tissue engineering. Among them, gold nanoparticles responding to changes in their surrounding environment are of particular interest due to their size-related optical properties. Here, we present a novel strategy for the preparation of gold nanoparticles exhibiting a stimuli-responsive behavior. We rely on the use of a ligand consisting of only a single repeat of the elastin-based pentapeptide VPGVG. In this contribution, we describe a protocol for the solid-phase peptide synthesis of thiol-terminated VPGVG ligand, and for the preparation of gold nanoparticles covered with the pentapeptide through a ligand-exchange reaction.

Electrostatic assembly of binary nanoparticle superlattices using protein cages.

Binary nanoparticle superlattices are periodic nanostructures with lattice constants much shorter than the wavelength of light and could be used to prepare multifunctional metamaterials. Such superlattices are typically made from synthetic nanoparticles, and although biohybrid structures have been developed, incorporating biological building blocks into binary nanoparticle superlattices remains challenging. Protein-based nanocages provide a complex yet monodisperse and geometrically well-defined hollow cage that can be used to encapsulate different materials. Such protein cages have been used to program the self-assembly of encapsulated materials to form free-standing crystals and superlattices at interfaces or in solution. Here, we show that electrostatically patchy protein cages--cowpea chlorotic mottle virus and ferritin cages--can be used to direct the self-assembly of three-dimensional binary superlattices. The negatively charged cages can encapsulate RNA or superparamagnetic iron oxide nanoparticles, and the superlattices are formed through tunable electrostatic interactions with positively charged gold nanoparticles. Gold nanoparticles and viruses form an AB(8)(fcc) crystal structure that is not isostructural with any known atomic or molecular crystal structure and has previously been observed only with large colloidal polymer particles. Gold nanoparticles and empty or nanoparticle-loaded ferritin cages form an interpenetrating simple cubic AB structure (isostructural with CsCl). We also show that these magnetic assemblies provide contrast enhancement in magnetic resonance imaging.

Using light and a molecular switch to 'lock' and 'unlock' the Diels-Alder reaction.

Light is used to 'gate' the Diels-Alder reaction using a photoresponsive dithienylfuran backbone and turn the reversibility of the Diels-Alder reaction 'off' and 'on' at 100 °C. These features make the reported system an excellent candidate for developing the next generation of self-healing polymers and photothermal drug delivery vehicles.

Elastin-based stimuli-responsive gold nanoparticles.

Gold nanoparticles covered with ligands consisting of only a single repeat of the elastin-based pentapeptide VPGVG exhibit a stimuli-responsive behaviour.

Reactivity-gated photochromism of 1,2-dithienylethenes for potential use in dosimetry applications.

[reaction: see text] A novel molecular switching system based on reactivity-gated photochromism operates because a butadiene undergoes a [4 + 2] cycloaddition reaction with a dienophile to produce the photoresponsive 1,2-dithienylethene backbone. The reversible change in color when samples are irradiated with appropriate wavelengths of light occurs only after the Diels-Alder cycloaddition reaction takes place.