Late cardiac sodium current can be assessed using automated patch-clamp

The cardiac late Na (+) current is generated by a small fraction of voltage-dependent Na (+) channels that undergo a conformational change to a burst-gating mode, with repeated openings and closures during the action potential (AP) plateau. Its magnitude can be augmented by inactivation-defective mutations, myocardial ischemia, or prolonged exposure to chemical compounds leading to drug-induced (di)-long QT syndrome, and results in an increased susceptibility to cardiac arrhythmias. Using CytoPatch™ 2 automated patch-clamp equipment, we performed whole-cell recordings in HEK293 cells stably expressing human Nav1.5, and measured the late Na (+) component as average current over the last 100 ms of 300 ms depolarizing pulses to -10 mV from a holding potential of -100 mV, with a repetition frequency of 0.33 Hz. Averaged values in different steady-state experimental conditions were further corrected by the subtraction of current average during the application of tetrodotoxin (TTX) 30 μM. We show that ranolazine at 10 and 30 μM in 3 min applications reduced the late Na (+) current to 75.0 ± 2.7% (mean ± SEM, n = 17) and 58.4 ± 3.5% ( n = 18) of initial levels, respectively, while a 5 min application of veratridine 1 μM resulted in a reversible current increase to 269.1 ± 16.1% ( n = 28) of initial values. Using fluctuation analysis, we observed that ranolazine 30 μM decreased mean open probability p from 0.6 to 0.38 without modifying the number of active channels n, while veratridine 1 μM increased n 2.5-fold without changing p. In human iPSC-derived cardiomyocytes, veratridine 1 μM reversibly increased APD90 2.12 ± 0.41-fold (mean ± SEM, n = 6). This effect is attributable to inactivation removal in Nav1.5 channels, since significant inhibitory effects on hERG current were detected at higher concentrations in hERG-expressing HEK293 cells, with a 28.9 ± 6.0% inhibition (mean ± SD, n = 10) with 50 μM veratridine.

Catch Rates, Composition and Fish Size from Reefs Managed with Periodically-Harvested Closures

Periodically-harvested closures are commonly employed within co-management frameworks to help manage small-scale, multi-species fisheries in the Indo-Pacific. Despite their widespread use, the benefits of periodic harvesting strategies for multi-species fisheries have, to date, been largely untested. We examine catch and effort data from four periodically-harvested reef areas and 55 continuously-fished reefs in Solomon Islands. We test the hypothesis that fishing in periodically-harvested closures would yield: (a) higher catch rates, (b) proportionally more short lived, fast growing, sedentary taxa, and (c) larger finfish and invertebrates, compared to catches from reefs continuously open to fishing. Our study showed that catch rates were significantly higher from periodically-harvested closures for gleaning of invertebrates, but not for line and spear fishing. The family level composition of catches did not vary significantly between open reefs and periodically-harvested closures. Fish captured from periodically-harvested closures were slightly larger, but Trochus niloticus were significantly smaller than those from continuously open reefs. In one case of intense and prolonged harvesting, gleaning catch rates significantly declined, suggesting invertebrate stocks were substantially depleted in the early stages of the open period. Our study suggests periodically-harvested closures can have some short term benefits via increasing harvesting efficiency. However, we did not find evidence that the strategy had substantially benefited multi-species fin-fisheries.

Academic cooperation to foster research and advocacy competences in the occupied Palestinian territory (West Bank)

Palestinians living in the West Bank, a territory occupied by the State of Israel according to International Law, face deprived access to land and a limited ability to move freely which pertains to the presence of Israeli settlements and other infrastructure (closures, restricted or forbidden roads, etc.). This confinement has significant impacts on their economic and social livelihoods, and it is even worsening with the on-going construction of a 709 km long Barrier which mainly runs inside the West Bank. With regard to this situation, there is a clear need to strengthen the capacity of civil society and its representatives to apply sound research processes as a basis for improved advocacy for Palestinian human rights. Monitoring processes and tools are needed to assess the impacts of the Palestinians’ confinement, particularly in relation to the Barrier’s construction. Reliable data has also to be collected, managed, and above all, shared. These challenges have been addressed within the Academic Cooperation Palestine Project (ACPP) that brings together academic partners from the occupied Palestinian territory (oPt) West Bank (WB), and Switzerland as well as other international academic institutions and Palestinian governmental and non-governmental agencies. ACPP started in early 2011 and is designed as a large cooperation networking platform involving researchers, students, public servants and experts from the oPt WB. A large set of actions have already been developed during the first year of the project, including courses, training, and research actions. First relevant results and impacts of the different actions are presented in this paper. Taken as a whole, the project produces valuable results for all partners: useful advocacy material for the Palestinian partners, and a unique “real-scale laboratory” where investigations are jointly conducted to develop novel confinement and change indicators.

Long-term trends in invertebrate–habitat relationships under protected and fished conditions

Few studies examine the long-term effects of changing predator size and abundance on the habitat associations of resident organisms despite that this knowledge is critical to understand the ecosystem effects of fishing. Marine reserves offer the opportunity to determine ecosystem-level effects of manipulated predator densities, while parallel monitoring of adjacent fished areas allows separating these effects from regional-scale change. Relationships between two measures of benthic habitat structure (reef architecture and topographic complexity) and key invertebrate species were followed over 17 years at fished and protected subtidal rocky reefs associated with two southern Australian marine reserves. Two commercially harvested species, the southern rock lobster (Jasus edwardsii) and blacklip abalone (Haliotis rubra) were initially weakly associated with habitat structure across all fished and protected sites. The strength of association with habitat for both species increased markedly at protected sites 2 years after marine reserve declaration, and then gradually weakened over subsequent years. The increasing size of rock lobster within reserves apparently reduced their dependency on reef shelters as refuges from predation. Rising predation by fish and rock lobster in the reserves corresponded with weakening invertebrate–habitat relationships for H. rubra and sea urchins (Heliocidaris erythrogramma). These results emphasise that animal–habitat relationships are not necessarily stable through time and highlight the value of marine reserves as reference sites. Our work shows that fishery closures to enhance populations of commercially important and keystone species should be in areas with a range of habitat features to accommodate shifting ecological requirements with ontogenesis.

A new technique to automatically quantify microstructures of fine grained carbonate mylonites: two-step etching combined with SEM imaging and image analysis

A two-step etching technique for fine-grained calcite mylonites using 0.37% hydrochloric and 0.1% acetic acid produces a topographic relief which reflects the grain boundary geometry. With this technique, calcite grain boundaries become more intensely dissolved than their grain interiors but second phase minerals like dolomite, quartz, feldspars, apatite, hematite and pyrite are not affected by the acid and therefore form topographic peaks. Based on digital backscatter electron images and element distribution maps acquired on a scanning electron microscope, the geometry of calcite and the second phase minerals can be automatically quantified using image analysis software. For research on fine-grained carbonate rocks (e.g. dolomite calcite mixtures), this low-cost approach is an attractive alternative to the generation of manual grain boundary maps based on photographs from ultra-thin sections or orientation contrast images.