Selective Duplex Formation in Mixed Sequence Libraries of Synthetic Polymers.

Recognition-encoded melamine oligomers (REMO) are synthetic polymers that feature an alternating 1,3,5-triazine-piperazine backbone and side-chains equipped with either a phenol or phosphine oxide recognition unit. An automated method for the solid-phase synthesis (SPS) of REMO of any specified sequence has been developed starting from dichlorotriazine monomer building blocks. Complementary homo-oligomers with either six phenols or six phosphine oxides were synthesized and shown to form a stable duplex in nonpolar solvents by NMR denaturation experiments. The duplex was covalently trapped by equipping the ends of the oligomers with an azide and an alkyne group and using a copper-catalyzed alkyne-azide cycloaddition (CuAAC) reaction. The SPS methodology was adapted to synthesize mixed sequence libraries by using a mixture of two different dichlorotriazine building blocks in each coupling cycle of an oligomer synthesis. The resulting libraries contain statistical mixtures of all possible sequences. The self-assembly properties of these libraries were screened by using the CuAAC reaction to trap any duplexes present. In mixed sequence libraries of 6-mers, the trapping experiments showed that only sequence-complementary oligomers formed duplexes at micromolar concentrations in dichloromethane. The automated synthesis approach developed here provides access to large libraries of mixed sequence synthetic polymers, and the covalent trapping experiment provides a convenient tool for screening functional properties of mixtures. The results suggest high-fidelity sequence-selective duplex formation in mixtures of 6-mer sequences of the REMO architecture.

Timing is everything; operational changes at a pumping station with a gravity sluice to provide safe downstream passage for silver European eels and deliver considerable financial savings.

Catadromous European eel (Anguilla anguilla) are a critically endangered fish species due in part to in-river anthropogenic barriers (e.g., pumping stations, weirs, hydropower facilities). European legislation stipulates that safe downstream passage must be provided at hazardous intakes. Where present, gravity sluices have the potential to act as safe and low-cost downstream passage for seaward migrating silver eels at pumping station, but operational changes are required. This study used catchment-wide and fine-scale acoustic telemetry to investigate if operational changes (OC) at a pumping station (PS) with a co-located gravity sluice (GS) facilitated safe downstream passage for silver European eels. Specifically, night-time pump operations were ceased, river levels prior to sluicing were elevated and the GS was opened during key eel migration windows, i.e., at night during the new moon phase in autumn. No tagged eels passed through any pumps and the majority (2018 = 87.5%, 2020 = 88.9%) that approached the PS during OC passed downstream through the GS. Most eels approached during the first period of night sluicing after release (2018 = 73.9% and 2020 = 76.5%) and passed downstream during the first sluice event they experienced at the PS (2018 = 66.7% and 2020 = 75.0%). During the final approach prior to passage, very few retreats back upstream occurred at a median (IQR) distance of 34 (7.25) m from the GS and were predominantly a short distance (1-8 m). Overall, OC at a PS with a GS are considered a win-win-win, despite opening the sluice for <3% of the study period, given safe downstream passage was maximised, the financial benefits of sluicing water (∼£14,670 in direct operational costs over two years) and the relative ease of implementation.

Rethinking fish-friendliness of pumps by shifting focus to both safe and timely fish passage for effective conservation.

Globally, catadromous freshwater eels of the genus Anguilla are of conservation concern, including critically endangered European eel (Anguilla anguilla). Pumping stations that move river water to a higher elevation severely impact eels during their seaward spawning migration. Fish-friendly pumps can mitigate fish injury and mortality but here we uniquely rethink a fish-friendly pump as a fish passage solution. In this pluriannual study, the seasonal timing of pump operation was misaligned with the typical silver eel migration period. Eels were almost exclusively nocturnal but night-time pumping represented as little as 5.6% a year. Night-time eel approaches were primarily influenced by pump duration and temperature, but did not align with lunar phase, unlike in unregulated rivers. After reaching the pumping station, eel passage was influenced by weedscreen aperture and increased when the aperture was increased. Passive sensor collision suggested non-pump infrastructure could cause injury and mortality to eels. It is therefore recommended pump operation should align with the timing of silver eel migration, weedscreen and pump entrance efficiencies should be maximised, and non-pump infrastructure must have low fish injury risk. Ultimately, considering the entire structure a fish passage solution will help ensure fish-friendly pumps have high conservation value for anguillid eels globally.

An automated paradigm for Drosophila visual psychophysics.

BACKGROUND: Mutations that cause learning and memory defects in Drosophila melanogaster have been found to also compromise visual responsiveness and attention. A better understanding of attention-like defects in such Drosophila mutants therefore requires a more detailed characterization of visual responsiveness across a range of visual parameters. METHODOLOGY/PRINCIPAL FINDINGS: We designed an automated behavioral paradigm for efficiently dissecting visual responsiveness in Drosophila. Populations of flies walk through multiplexed serial choice mazes while being exposed to moving visuals displayed on computer monitors, and infra-red fly counters at the end of each maze automatically score the responsiveness of a strain. To test our new design, we performed a detailed comparison between wild-type flies and a learning and memory mutant, dunce(1). We first confirmed that the learning mutant dunce(1) displays increased responsiveness to a black/green moving grating compared to wild type in this new design. We then extended this result to explore responses to a wide range of psychophysical parameters for moving gratings (e.g., luminosity, contrast, spatial frequency, velocity) as well as to a different stimulus, moving dots. Finally, we combined these visuals (gratings versus dots) in competition to investigate how dunce(1) and wild-type flies respond to more complex and conflicting motion effects. CONCLUSIONS/SIGNIFICANCE: We found that dunce(1) responds more strongly than wild type to high contrast and highly structured motion. This effect was found for simple gratings, dots, and combinations of both stimuli presented in competition.