Climate and human stressors on global penguin hotspots: Current assessments for future conservation.

As charismatic and iconic species, penguins can act as "ambassadors" or flagship species to promote the conservation of marine habitats in the Southern Hemisphere. Unfortunately, there is a lack of reliable, comprehensive, and systematic analysis aimed at compiling spatially explicit assessments of the multiple impacts that the world's 18 species of penguin are facing. We provide such an assessment by combining the available penguin occurrence information from Global Biodiversity Information Facility (>800,000 occurrences) with three main stressors: climate-driven environmental changes at sea, industrial fisheries, and human disturbances on land. Our analyses provide a quantitative assessment of how these impacts are unevenly distributed spatially within species' distribution ranges. Consequently, contrasting pressures are expected among species, and populations within species. The areas coinciding with the greatest impacts for penguins are the coast of Perú, the Patagonian Shelf, the Benguela upwelling region, and the Australian and New Zealand coasts. When weighting these potential stressors with species-specific vulnerabilities, Humboldt (Spheniscus humboldti), African (Spheniscus demersus), and Chinstrap penguin (Pygoscelis antarcticus) emerge as the species under the most pressure. Our approach explicitly differentiates between climate and human stressors, since the more achievable management of local anthropogenic stressors (e.g., fisheries and land-based threats) may provide a suitable means for facilitating cumulative impacts on penguins, especially where they may remain resilient to global processes such as climate change. Moreover, our study highlights some poorly represented species such as the Northern Rockhopper (Eudyptes moseleyi), Snares (Eudyptes robustus), and Erect-crested penguin (Eudyptes sclateri) that need internationally coordinated efforts for data acquisition and data sharing to understand their spatial distribution properly.

The breeding biology of erect-crested penguins, Eudyptes sclateri: Hormones, behavior, obligate brood reduction and conservation.

Erect-crested penguins are the least studied of all penguins. They breed on two isolated subantarctic island groups, the Antipodes and Bounty Islands. Sporadic nest counts indicate a dramatic decline in numbers of erect-crested penguins over the last 50 years. Here we present data from a study undertaken in 1998 on the breeding biology, behavior and hormones of erect-crested penguins. It represents, even today, by far the most detailed data available on this species. The penguins exhibited extreme reversed egg-size dimorphism, whereby the first-laid A-egg was much smaller than the second-laid B-egg. A-eggs were lost before (42.3%) or on (37.8%) the day the B-egg was laid, and none survived more than 7 days after that. The penguins were in a low state of reproductive readiness, as evidenced by low levels of copulation, fighting, and testosterone in males during the courtship/laying period when, curiously, plasma levels of testosterone were at least as high in females. The laying interval (5.4 days) is the longest recorded for any penguin species, and incubation was highly variable until clutch completion. Most nests (91.2%) contained no nesting material and eggs were laid directly onto the ground. A-eggs were lost mainly by rolling out of the nest. However, even when prevented from doing so by an experimental manipulation, A-eggs survived no longer than those in control nests. Testosterone levels in males increased after clutch completion, when they remained in attendance at the nest for up to 13 days, despite females assuming most of the incubation duties. The bills of males were significantly larger than those of females and probably help with guarding the nest. We discuss explanations for obligate brood reduction in crested penguins and the options for conservation in light of our census results, which indicate that this enigmatic penguin species could be in trouble.

Towards the protection of ammunition headstamps during fingermark enhancement processing; a preliminary study.

Forensic recovery from fired ammunition casings remains one of the most challenging tasks during high-profile investigations. Often, the decision must be made between screening for DNA or fingerprints, and, in doing so, the impact these processes will have on the examination of ballistic markings imparted on the ammunition from the firearm itself. Traditionally, fingermark enhancement processes have yielded low success rates in their efforts to identify suspects by enhancing friction ridge detail left on the cartridge casings. Moreover, the enhancement methods utilised may often induce detrimental physical changes to the casing(s), rendering them unsuitable for subsequent ballistics (marking) examination. Recently, new technology has been shown to increase the success rate of fingermark recovery from fired ammunition, and the growing adoption of such innovation means that new challenges are encountered to maximise evidence recovery and streamline forensic workflows. One such example arises from the potential obscuration of the ammunition headstamp area during such treatments. Accordingly, this study outlines the preliminary investigations and developments of a polymer mask substrate that serves to protect the headstamp of fired ammunition casings during relevant fingermark enhancement processes. The technique also has the capacity to be used as a surface protection device to eliminate unwanted chemical deposition across other areas of interest and evidence types.

Using a quadruplet codon to expand the genetic code of an animal.

Genetic code expansion in multicellular organisms is currently limited to the use of repurposed amber stop codons. Here, we introduce a system for the use of quadruplet codons to direct incorporation of non-canonical amino acids in vivo in an animal, the nematode worm Caenorhabditis elegans. We develop hybrid pyrrolysyl tRNA variants to incorporate non-canonical amino acids in response to the quadruplet codon UAGA. We demonstrate the efficiency of the quadruplet decoding system by incorporating photocaged amino acids into two proteins widely used as genetic tools. We use photocaged lysine to express photocaged Cre recombinase for the optical control of gene expression and photocaged cysteine to express photo-activatable caspase for light inducible cell ablation. Our approach will facilitate the routine adoption of quadruplet decoding for genetic code expansion in eukaryotic cells and multicellular organisms.

Remote magnetic navigation compared to contemporary manual techniques for the catheter ablation of ventricular arrhythmias in structural heart disease.

BACKGROUND: There are limited data comparing remote magnetic navigation (RMN) to contemporary techniques of manual-guided ventricular arrhythmia (VA) catheter ablation. OBJECTIVES: We compared acute and long-term outcomes of VA ablation guided by either RMN or contemporary manual techniques in patients with structural heart disease. METHODS: From 2010-2019, 192 consecutive patients, with ischemic cardiomyopathy (ICM) or non-ischemic cardiomyopathy (NICM) underwent catheter ablation for sustained ventricular tachycardia (VT) or premature ventricular complexes (PVCs), using either RMN (n = 60) or manual (n = 132) guided techniques. Acute success and VA-free survival were compared. RESULTS: In ICM, acute procedural success was comparable between the 2 techniques (manual 43.5% vs. RMN 29%, P = 0.11), as was VA-free survival (manual 83% vs. RMN 74%, P = 0.88), and survival free from cardiac transplantation and all-cause mortality (manual 88% vs. RMN 87%, P = 0.47), both at 12-months after final ablation. In NICM, manual compared to RMN guided, had superior acute procedural success (manual 46% vs. RMN 19%, P = 0.003) and VA-free survival 12-months after final ablation (manual 79% vs. RMN 41%, P = 0.004), but comparable survival free from cardiac transplantation and all-cause mortality 12-months after final ablation (manual 95% vs. RMN 90%, P = 0.52). Procedural duration was shorter in both subgroups undergoing manual guided ablation, whereas fluoroscopy dose and complication rates were comparable. CONCLUSION: RMN provides similar outcomes to manual ablation in patients with ICM. In NICM however, acute success, and long-term VA-free survival was better with manual ablation. Prospective, multi-centre randomised trials comparing contemporary manual and RMN systems for VA catheter ablation are needed.

Axillary vein access using ultrasound guidance, Venography or Cephalic Cutdown-What is the optimal access technique for insertion of pacing leads?

We reviewed the different approaches used for central vein access during insertion of cardiac implantable electronic devices. The benefits and hazards of each approach (cephalic vein cutdown, axillary vein cannulation using venography and ultrasound) are discussed. Each approach has its advantages and hazards that need to be considered for the individual patient and balanced against the skills of the operator. The benefits of ultrasound guided venous access in reducing radiation exposure to the patient and implanter, avoiding the need for angiographic contrast and in minimizing the risk of pneumothorax and inadvertent arterial puncture are highlighted. Trainees should be taught each approach to deal with patient variability. Ultrasound guidance should be considered as a mainstream option for most patients.

Precise optical control of gene expression in C elegans using improved genetic code expansion and Cre recombinase.

Synthetic strategies for optically controlling gene expression may enable the precise spatiotemporal control of genes in any combination of cells that cannot be targeted with specific promoters. We develop an improved genetic code expansion system in Caenorhabditis elegans and use it to create a photoactivatable Cre recombinase. We laser-activate Cre in single neurons within a bilaterally symmetric pair to selectively switch on expression of a loxP-controlled optogenetic channel in the targeted neuron. We use the system to dissect, in freely moving animals, the individual contributions of the mechanosensory neurons PLML/PLMR to the C. elegans touch response circuit, revealing distinct and synergistic roles for these neurons. We thus demonstrate how genetic code expansion and optical targeting can be combined to break the symmetry of neuron pairs and dissect behavioural outputs of individual neurons that cannot be genetically targeted.

Animal behaviour and movement emerges from the stimulation of nerve cells that are connected together like a circuit. Researchers use various tools to investigate these neural networks in model organisms such as roundworms, fruit flies and zebrafish. The trick is to activate some nerve cells, but not others, so as to isolate their specific role within the neural circuit. One way to do this is to switch genes on or off in individual cells as a way to control their neuronal activity. This can be achieved by building a photocaged version of the enzyme Cre recombinase which is designed to target specific genes. The modified Cre recombinase contains an amino acid (the building blocks of proteins) that inactivates the enzyme. When the cell is illuminated with UV light, a part of the amino acid gets removed allowing Cre recombinase to turn on its target gene. However, cells do not naturally produce these photocaged amino acids. To overcome this, researchers can use a technology called genetic code expansion which provides cells with the tools they need to build proteins containing these synthetic amino acids. Although this technique has been used in live animals, its application has been limited due to the small amount of proteins it produces. Davis et al. therefore set out to improve the efficiency of genetic code expansion so that it can be used to study single nerve cells in freely moving roundworms. In the new system, named LaserTAC, individual cells are targeted with UV light that 'uncages' the Cre recombinase enzyme so it can switch on a gene for a protein that controls neuronal activity. Davis et al. used this approach to stimulate a pair of neurons sensitive to touch to see how this impacted the roundworm's behaviour. This revealed that individual neurons within this pair contribute to the touch response in different ways. However, input from both neurons is required to produce a robust reaction. These findings show that the LaserTAC system can be used to manipulate gene activity in single cells, such as neurons, using light. It allows researchers to precisely control in which cells and when a given gene is switched on or off. Also, with the improved efficiency of the genetic code expansion, this technology could be used to modify proteins other than Cre recombinase and be applied to other artificial amino acids that have been developed in recent years.

Diffusivity Measurement by Single-Molecule Recycling in a Capillary Microchannel.

Microfluidic devices have been extensively investigated in recent years in fields including ligand-binding analysis, chromatographic separation, molecular dynamics, and DNA sequencing. To prolong the observation of a single molecule in aqueous buffer, the solution in a sub-micron scale channel is driven by a electric field and reversed after a fixed delay following each passage, so that the molecule passes back and forth through the laser focus and the time before irreversible photobleaching is extended. However, this practice requires complex chemical treatment to the inner surface of the channel to prevent unexpected sticking to the surface and the confined space renders features, such as a higher viscosity and lower dielectric constant, which slow the Brownian motion of the molecule compared to the bulk solution. Additionally, electron beam lithography used for the fabrication of the nanochannel substantially increases the cost, and the sub-micron dimensions make the molecule difficult to locate. In this paper, we propose a method of single-molecule recycling in a capillary microchannel. A commercial fused-silica capillary with an inner diameter of 2 microns is chopped into a 1-inch piece and is fixed onto a cover slip. Two o-rings on the sides used as reservoirs and an o-ring in the middle used as observation window are glued over the capillary. The inner surface of the capillary is chemically processed to reduce the non-specific sticking and to improve capillary effect. The device does not require high-precision fabrication and thus is less costly and easier to prepare than the nanochannel. 40 nm Fluospheres® in 50% methanol are used as working solution. The capillary is translated by a piezo stage to recycle the molecule, which diffuses freely through the capillary, and a confocal microscope is used for fluorescence collection. The passing times of the molecule through the laser focus are calculated by a real-time control system based on an FPGA, and the commands of translation are given to the piezo stage through a feedback algorithm. The larger dimensions of the capillary overcomes the strong sticking, the reduced diffusivity, and the difficulty of localizing the molecule. We have achieved a maximum number of recycles of more than 200 and developed a maximum-likelihood estimation of the diffusivity of the molecule, which attains results of the same magnitude as the previous report. This technique simplifies the overall procedure of the single-molecule recycling and could be useful for the ligand-binding studies in high-throughput screening.

Visualising natural attractions within national parks: Preferences of tourists for photographs with different visual characteristics.

To explore what types of photographs are more helpful means to interpret natural attractions within national parks, this study focused on the relationship between the photographs with different visual characteristics and their perceived visual appeal. A photograph-based Q method was adopted. Results confirmed the visual quality of a photograph was the most important characteristic that determined its perceived attractiveness; those photographs with a high visual quality could successfully attract tourists' attention. The subject also significantly affected the preferences of observers, suggesting an interest-dependent pattern. Using photographs of birds as examples, the participants who were interested in birds were attracted by the photographs of birds rather than those of other subjects. This study provides a better understanding of the effectiveness of photographs for communication. Findings may help researchers, communicators and national park marketers better understand and select appropriate photographs for interpretation within national parks.

A picture is not always worth a thousand words: The visual quality of photographs affects the effectiveness of interpretive signage for science communication.

Photographs are widely used to visualise science. However, the influence of visual quality of photographs on the effectiveness of science communication is often disregarded and seldom tested. This study examines how the visual quality of photographs affects science communication on interpretive signage within Xixi National Wetland Park, China. Results show that a high-quality photograph on the signage significantly increased the following affective and cognitive aspects of communication: (1) a visitor's intention to read the signage, (2) enjoyment of reading the interpretive information, (3) understanding and (4) recall of the information on the signage. This study offers empirical evidence for the benefits of using high-quality photographs to visualise science. To improve the effectiveness of science communication in interpretive products, we argue that the quality of the selected photographs should be considered seriously.

Transformation of the media landscape: Infotainment versus expository narrations for communicating science in online videos.

Society is undergoing a transformation in the way people consume media: increasingly we are using online on-demand videos, with the fastest growing segment of online videos about science being user-generated content that uses an infotainment style of delivery, in contrast to the traditional expository narrations of professionally generated content. In this study, we produced two otherwise identical videos about climate change to test the effects of an infotainment or expository narration. A total of 870 survey participants (419 English; 451 Spanish) were randomly presented with either an infotainment or expository version of the video. The expository narration was liked and believed more, and this held irrespective of language, age, sex or online viewing habits. However, the infotainment version was liked more by viewers without a university education and, further, viewers were better able to recall information from it, suggesting that user-generated content with infotainment-style narrations may actually be good for increasing public understanding of science.

Three-dimensional feedback-driven trapping of a single nanoparticle or molecule in aqueous solution with a confocal fluorescence microscope.

Control of electroosmotic flows in a two-layer microfluidic device with crossed channels is used to counteract Brownian diffusion in aqueous solution for three-dimensional trapping of a single nanoparticle or molecule within the probe volume of a confocal fluorescence microscope. A field programmable gate array sorts and counts photons into four channels synchronous with laser pulses in four beams focused to waists slightly offset from the center of the confocal volume and uses the counts to update voltages between the four fluidic inlets every 13.5 µs. Trapping is demonstrated for 40 nm nanoparticles for up to 240 s, 20 nm nanoparticles for up to 25 s, and single molecules of streptavidin-Alexa 647 for up to 1.2 s.

The use of longwave reflected UV imaging for the enhancement of cyanoacrylate developed fingermarks: A simple, safe and effective imaging tool.

Longwave ultraviolet reflection (LWUVR) imaging is reported as a simple, safe and non-invasive technique that significantly aids in the visualisation of cyanoacrylate developed latent fingermarks. The process can precede traditional cyanoacrylate dye staining and often removes the necessity to use these chemical contrast reagents, saving time, cost and eliminating mess. The non-destructive nature of the process and high resolution images that are obtained, builds upon issues that surround shorter-wavelength UV imaging. It has been demonstrated that, for most samples tested (a range of non-porous and semi-porous evidence), LWUVR imaging provided superior or similar results to those obtained using the traditional BY40 dye stain. The lack of penetration depth by LWUV radiation means that only the features of the surface under observation is detected by the camera, meaning that interference that may otherwise arise from fluorescence on the reverse side of the evidence (BY40 or fluorescent inks) under conventional fluorescent dye stain imaging modes, is mitigated. A new sequential processing workflow is proposed that does not impede with the conventional and widely adopted fume>stain>fluorescence sequence, but in fact allows LWUVR imaging to be conducted in a manner that serves to benefit the sequence and, ideally, save time during the examination and treatment of evidence.

Early and long-term outcomes after manual and remote magnetic navigation-guided catheter ablation for ventricular tachycardia.

Aims: Remote magnetic navigation (RMN) is a safe and effective means of performing ventricular tachycardia (VT) ablation. It may have advantages over manual catheter ablation due to ease of manoeuvrability and catheter stability. We sought to compare the safety and efficacy of RMN vs. manual VT ablation. Methods and results: Retrospective study of procedural outcomes of 139 consecutive VT ablation procedures (69 RMN, 70 manual ablation) in 113 patients between 2009 and 2015 was performed. Remote magnetic navigation was associated with overall higher acute procedural success (80% vs. 60%, P = 0.01), with a trend to fewer major complications (3% vs. 9% P = 0.09). Seventy-nine patients were followed up for a median of 17.0 [interquartile range (IQR) 3.0-41.0] months for the RMN group and 15.5 (IQR 6.5-30.0) months for manual ablation group. In the ischaemic cardiomyopathy subgroup, RMN was associated with longer survival from the composite endpoint of VT recurrence leading to defibrillator shock, re-hospitalization or repeat catheter ablation and all-cause mortality; single-procedure adjusted hazard ratio (HR) 0.240 (95% CI 0.070-0.821) P = 0.023, multi-procedure HR 0.170 (95% CI 0.046-0.632) P = 0.002. In patients with implanted defibrillators, multi-procedure VT-free survival was superior with RMN, HR 0.199 (95% CI 0.060-0.657) P = 0.003. Conclusion: Remote magnetic navigation may improve clinical outcomes after catheter ablation of VT in patients with ischaemic cardiomyopathy. Further prospective clinical studies are required to confirm these findings.

Genetic Encoding of Unnatural Amino Acids in C. elegans.

Site-specific incorporation of unnatural amino acids (UAAs) has greatly expanded the toolkit available to study biological phenomena in single cells. However, to address questions involving complex cellular interactions such as development, ageing, and the functions of the nervous system it is often necessary to use multicellular model organisms. The nematode Caenorhabditis elegans was the first organism to have its genetic code expanded. Due to its small size, ease of cultivation, and excellent UAA incorporation efficiency, C. elegans makes an ideal model organism to apply UAAs as tools to investigate the functioning of multicellular systems.Here, we describe methods to generate transgenic C. elegans capable of UAA incorporation, as well as how to deliver unnatural amino acids and test incorporation. Furthermore, we describe methods to uncage photosensitive unnatural amino acid derivatives.

Climate impacts on global hot spots of marine biodiversity.

Human activities drive environmental changes at scales that could potentially cause ecosystem collapses in the marine environment. We combined information on marine biodiversity with spatial assessments of the impacts of climate change to identify the key areas to prioritize for the conservation of global marine biodiversity. This process identified six marine regions of exceptional biodiversity based on global distributions of 1729 species of fish, 124 marine mammals, and 330 seabirds. Overall, these hot spots of marine biodiversity coincide with areas most severely affected by global warming. In particular, these marine biodiversity hot spots have undergone local to regional increasing water temperatures, slowing current circulation, and decreasing primary productivity. Furthermore, when we overlapped these hot spots with available industrial fishery data, albeit coarser than our estimates of climate impacts, they suggest a worrying coincidence whereby the world's richest areas for marine biodiversity are also those areas mostly affected by both climate change and industrial fishing. In light of these findings, we offer an adaptable framework for determining local to regional areas of special concern for the conservation of marine biodiversity. This has exposed the need for finer-scaled fishery data to assist in the management of global fisheries if the accumulative, but potentially preventable, effect of fishing on climate change impacts is to be minimized within areas prioritized for marine biodiversity conservation.

Visualisation of latent fingermarks on polymer banknotes using copper vacuum metal deposition: A preliminary study.

The UK's recent move to polymer banknotes has seen some of the currently used fingermark enhancement techniques for currency potentially become redundant, due to the surface characteristics of the polymer substrates. Possessing a non-porous surface with some semi-porous properties, alternate processes are required for polymer banknotes. This preliminary investigation explored the recovery of fingermarks from polymer notes via vacuum metal deposition using elemental copper. The study successfully demonstrated that fresh latent fingermarks, from an individual donor, could be clearly developed and imaged in the near infrared. By varying the deposition thickness of the copper, the contrast between the fingermark minutiae and the substrate could be readily optimised. Where the deposition thickness was thin enough to be visually indistinguishable, forensic gelatin lifters could be used to lift the fingermarks. These lifts could then be treated with rubeanic acid to produce a visually distinguishable mark. The technique has shown enough promise that it could be effectively utilised on other semi- and non-porous substrates.