Cloning, expression and purification of envelope proteins E1 and E2 of western equine encephalitis virus and potential use of them as antigens in immunoassays

The genes encoding envelope proteins E1 and E2 of western equine encephalitis virus (WEEV) were respectively cloned into a prokaryotic T7 RNA polymerase-regulated expression vector. The recombinant C-terminal 6xHis-tagged WEEV E1 and E2 were expressed in bacteria as inclusion bodies that were subsequently solubilized with 8M urea, purified by immobilized metal ion affinity chromatography and finally refolded using an arginine system. The purified 6xHis-tagged proteins showed 50kDa bands as revealed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, consistent with the expected sizes of WEEV E1 and E2. The potential of the recombinant WEEV E1 and E2 as antigens for serologic tests to detect anti-WEEV antibodies for diagnosis of WEEV infection was assessed by an enzyme-linked immunosorbent assay with anti-WEEV polyclonal antibodies obtained from the mice infected with WEEV. The anti-WEEV antibodies bound the recombinant WEEV E1 and E2 in a dose dependent manner. On the contrary, antibodies against Venezuelan equine encephalitis virus with a genetic background and a disease spectrum very similar to WEEV, did not bind to the recombinant WEEV E1 and E2. Our results suggest that the recombinant WEEV E1 and E2 possess predominant antigenicity of WEEV and have the potential to be used as antigens in immunoassays to detect anti-WEEV antibodies for serological diagnosis of WEEV infection so as to eliminate the need for preparation of cell culture-derived viral antigens, which is time-consuming, expensive, laborious, tedious, and hazardous.     

Nutrient dynamics in integrated aquaculture–hydroponics systems

Changes in concentrations of dissolved Ca, Cu, Fe, K, Mg, Mn, NO₃-N, Na, P, and Zn, as a function of increasing biomass of Nile tilapia (Oreochromis niloticus) fed a commercial diet in integrated aquaculture–hydroponic systems growing romaine lettuce (Lactuca sativa longifolia cv. Jericho), were monitored over three, 28-day experimental trials. The integrated systems, 320–330 l in volume, were filled initially with a complete nutrient hydroponic solution, and were operated at a 100% recirculation rate. Treatment levels were 0 (control) 151, 377, 902, and 1804 g of fish/system and treatments were conducted in duplicate. Nutrient concentrations and mutual ratios of nutrients quickly departed from initial conditions because the relative proportion of dissolved nutrients excreted by fish and subsequently absorbed by plants differed. The removal of nutrients by lettuce, fish, and solids collection was determined as a function of treatment size. The objective of this study was to quantify both the flow of nutrients through representative integrated aquaculture–hydroponics systems and the effects of different quantities of feed nutrient input on changes in nutrient-specific concentrations.     

Estimating density of a territorial species in a dynamic landscape

Context

Conservation planning for at-risk species requires understanding of where species are likely to occur, how many individuals are likely to be supported on a given landscape, and the ability to monitor those changes through time.

Objectives

We developed a distribution model for northern spotted owls that incorporates both habitat suitability and probability of territory occupancy while accounting for interspecies competition.

Methods

We developed range-wide habitat suitability maps for two time periods (1993 and 2012) for northern spotted owls that accounted for regional differences in habitat use and home range size. We used these maps for a long-term demographic monitoring study area to assess habitat change and estimate the number of potential territories based on available habitat for both time periods. We adjusted the number of potential territories using known occupancy rates to estimate owl densities for both time periods. We evaluated our range-wide habitat suitability model using independent survey data.

Results

Our range-wide habitat maps predicted areas suitable for territorial spotted owl presence well. On the demographic study area, the amount of habitat declined 19.7% between 1993 and 2012, while our estimate of the habitat-based carrying capacity declined from 150 to 146 territories. Estimated number of occupied territories declined from 94 to 57.

Conclusions

Conservation and recovery of at-risk species depends on understanding how habitat changes over time in response to factors such as wildfire, climate change, biological invasions, and interspecies competition, and how these changes influence species distribution. We demonstrate a model-based approach that provides an effective planning tool.

     

Wafer-scale graphene integrated circuit

A wafer-scale graphene circuit was demonstrated in which all circuit components, including graphene field-effect transistor and inductors, were monolithically integrated on a single silicon carbide wafer. The integrated circuit operates as a broadband radio-frequency mixer at frequencies up to 10 gigahertz. These graphene circuits exhibit outstanding thermal stability with little reduction in performance (less than 1 decibel) between 300 and 400 kelvin. These results open up possibilities of achieving practical graphene technology with more complex functionality and performance.     

Getting to know you: reputation and trust in a two-person economic exchange

Using a multiround version of an economic exchange (trust game), we report that reciprocity expressed by one player strongly predicts future trust expressed by their partner-a behavioral finding mirrored by neural responses in the dorsal striatum. Here, analyses within and between brains revealed two signals-one encoded by response magnitude, and the other by response timing. Response magnitude correlated with the "intention to trust" on the next play of the game, and the peak of these "intention to trust" responses shifted its time of occurrence by 14 seconds as player reputations developed. This temporal transfer resembles a similar shift of reward prediction errors common to reinforcement learning models, but in the context of a social exchange. These data extend previous model-based functional magnetic resonance imaging studies into the social domain and broaden our view of the spectrum of functions implemented by the dorsal striatum.