ω-Conotoxin GVIA mimetics that bind and inhibit neuronal Cav2.2 ion channels

The neuronal voltage-gated N-type calcium channel (Ca_v2.2) is a validated target for the treatment of neuropathic pain. A small library of anthranilamide-derived ω-Conotoxin GVIA mimetics bearing the diphenylmethylpiperazine moiety were prepared and tested using three experimental measures of calcium channel blockade. These consisted of a ¹²⁵I-ω-conotoxin GVIA displacement assay, a fluorescence-based calcium response assay with SH-SY5Y neuroblastoma cells, and a whole-cell patch clamp electrophysiology assay with HEK293 cells stably expressing human Ca_v2.2 channels. A subset of compounds were active in all three assays. This is the first time that compounds designed to be mimics of ω-conotoxin GVIA and found to be active in the ¹²⁵I-ω-conotoxin GVIA displacement assay have also been shown to block functional ion channels in a dose-dependent manner.

Factors affecting patch occupancy by the white-browed treecreeper Climacteris Affinis in an agricultural landscape in north-west Victoria, Australia

The survival of habitat-dependent fauna within agricultural mosaics depends on their ability to occupy remnant habitat patches and move through the modified landscape. In north-west Victoria, Australia, less than 10% of the pre-European extent of Selah Casuarina pauper woodland remains intact due to agricultural development. The White-browed Treecreeper Climacteris affinis, is a small, insectivorous passerine that, in this region, preferentially inhabits Selah woodland. To assess the ability of C. affinis to persist in an agricultural landscape, 30 woodland sites in the Millewa landscape (34°30'S, 141°30'E) were surveyed, and patterns of patch occupancy used to examine the influence of spatial characteristics, landscape context and grazing by stock on the suitability of remnants as C. affinis habitat. Sites occupied by C. affinis were larger and less likely to be grazed by stock than vacant patches. The area-dependency of patch occupancy represents a step-threshold: C. affinis were not detected in remnants with less than 18.5 ha of Selah woodland but above this threshold, density was not correlated with patch area. Measures of patch isolation, the existence of linking linear "corridors" and tree density were not reliable indicators of patch occupancy. The presence of the species in remnants entirely surrounded by agricultural land suggests they are capable of crossing up to 450 m of cultivated land to prospect for habitat. The extensive network of linear vegetation and the numerous small remnants and scattered trees appear to facilitate movements of C. affinis in this landscape. Increasing the size of existing remnants, creating new habitat to expand the area of occupancy and maintaining landscape connectivity are priorities for the long-term management of this threatened species.

Retrodicting patch use by foraging swans in a heterogeneous environment using a set of functional responses

Effective conservation of important bird areas requires insight in the number of birds an area can support, and how this carrying capacity changes with habitat modifications. When food depletion is the dominant mechanism of competition, it should in principle be possible to calculate the total time foragers can spend per patch from their functional response (intake rate as a function of food density). However, in the field there are likely to be factors modulating the functional response. In this study previously published results of experiments on captive Bewick's swans were used to obtain functional responses of swans digging for tubers of Fennel pondweed on different foraging substrates: sandy and clayey sediment, and in shallow and deep water. In a field study, four 250×250 m sections belonging to different types (sandy–shallow, clayey–shallow, sandy–deep and clayey–deep) were delineated. Here tubers were sampled with sediment corers in three years, both before and after swan exploitation in autumn, and swans were observed and mapped from a hide in two of these years. Giving-up tuber biomass densities varied among sections. Substitution of these giving-up densities in the derived patch-type-specific functional responses yielded the quitting net energy intake rates in the four sections. As expected from the marginal value theorem, the quitting net energy intake rates did not vary among sections. Moreover, the observed foraging pressure (total foraging time per area) per patch type was in quantitative agreement with the integrated functional responses. These results suggest that in spatially heterogeneous environments, patch exploitation by foragers can be predicted from their functional responses after accounting for foraging substrate.

Seagrass patch size affects fish responses to edges

1. Patch area and proximity of patch edge can influence ecological processes across patchy landscapes and may interact with each other. Different patch sizes have different amounts of core habitat, potentially affecting animal abundances at the edge and middle of patches. In this study, we tested if edge effects varied with patch size.

2. Fish were sampled in 10 various-sized seagrass patches (114–5934 m²) using a small (0·5 m²) push net in three positions within each patch: the seagrass edge, 2 m into a patch and in the middle of a patch.

3. The two most common species showed an interaction between patch size and the edge–interior difference in abundance. In the smallest patches, pipefish (Stigmatopora nigra) were at similar densities at the edge and interior, but with increasing patch size, the density at the edge habitat increased. For gobies (Nesogobius maccullochi), the pattern was exactly the opposite.

4. This is the first example from a marine system of how patch size can influence the magnitude and pattern of edge effects.

5. Both patch area and edge effects need to be considered in the development of conservation and management strategies for seagrass habitats.

Adaptive patch size determination for patch-based image completion

Patch-based image completion proceeds by iteratively filling the target (unknown) region by the best matching patches in the source image. In most existing such algorithms, the size of the patches is either fixed and specified by a default number or simply chosen to be inversely proportional to the spatial frequency. However, it is noted that the patch size affects how well the filled patch captures the local characteristics of the source image and thus the final completion accuracy. Thus in this paper we propose a new method to compute appropriate patch sizes for image completion to improve its performance. In particular, we formulate the patch size determination as an optimization problem that minimizes an objective function involving image gradients and distinct and homogenous features. Experimental results show that our method can provide a significant enhancement to patch-based image completion algorithms.

Does reduced mobility through fragmented landscapes explain patch extinction patterns for three honeyeaters?

1.Habitat loss and associated fragmentation are major drivers of biodiversity decline, and understanding how they affect population processes (e.g. dispersal) is an important conservation goal. In a large-scale test employing 10 × 10 km units of replication, three species of Australian birds, the fuscous honeyeater, yellow-tufted honeyeater and white-plumed honeyeater, responded differently to fragmentation. The fuscous and yellow-tufted honeyeaters are ‘decliners’ that disappeared from suitable habitat in landscapes where levels of tree-cover fell below critical thresholds of 17 and 8%, respectively. The white-plumed honeyeater is a ‘tolerant’ species whose likelihood of occurrence in suitable habitat was independent of landscape-level tree-cover. 2.To determine whether the absence of the two decliner species in low tree-cover landscapes can be explained by reduced genetic connectivity, we looked for signatures of reduced mobility and gene flow in response to fragmentation across agricultural landscapes in the Box-Ironbark region of north-central Victoria, Australia. 3.We compared patterns of genetic diversity and population structure at the regional scale and across twelve 100 km2 landscapes with different tree-cover extents. We used genetic data to test landscape models predicting reduced dispersal through the agricultural matrix. We tested for evidence of sex-biased dispersal and sex-specific responses to fragmentation. 4.Reduced connectivity may have contributed to the disappearance of the yellow-tufted honey-eater from low tree-cover landscapes, as evidenced by male bias and increased relatedness among males in low tree-cover landscapes and signals of reduced gene flow and mobility through the agricultural matrix. We found no evidence for negative effects of fragmentation on gene flow in the other decliner, the fuscous honeyeater, suggesting that undetected pressures act on this species. As expected, there was no evidence for decreased movement through fragmented landscapes for the tolerant white-plumed honeyeater. 5.We demonstrated effects of habitat loss and fragmentation (stronger patterns of genetic differentiation, increased relatedness among males) on the yellow-tufted honeyeater above the threshold at which probability of occurrence dropped. Increasing extent and structural connectivity of habitat should be an appropriate management action for this species and other relatively sedentary woodland specialist species for which it can be taken as representative.

Restoration of self-awareness of hypoglycemia in adults with long-standing type 1 diabetes: hyperinsulinemic-hypoglycemic clamp substudy results from the HypoCOMPaSS trial.

OBJECTIVE: Impaired awareness of hypoglycemia (IAH) and defective counterregulation significantly increase severe hypoglycemia risk in type 1 diabetes (T1D). We evaluated restoration of IAH/defective counterregulation by a treatment strategy targeted at hypoglycemia avoidance in adults with T1D with IAH (Gold score ≥4) participating in the U.K.-based multicenter HypoCOMPaSS randomized controlled trial. RESEARCH DESIGN AND METHODS: Eighteen subjects with T1D and IAH (mean ± SD age 50 ± 9 years, T1D duration 35 ± 10 years, HbA1c 8.1 ± 1.0% [65 ± 10.9 mmol/mol]) underwent stepped hyperinsulinemic-hypoglycemic clamp studies before and after a 6-month intervention. The intervention comprised the HypoCOMPaSS education tool in all and randomized allocation, in a 2 × 2 factorial study design, to multiple daily insulin analog injections or continuous subcutaneous insulin infusion therapy and conventional glucose monitoring or real-time continuous glucose monitoring. Symptoms, cognitive function, and counterregulatory hormones were measured at each glucose plateau (5.0, 3.8, 3.4, 2.8, and 2.4 mmol/L), with each step lasting 40 min with subjects kept blinded to their actual glucose value throughout clamp studies. RESULTS: After intervention, glucose concentrations at which subjects first felt hypoglycemic increased (mean ± SE from 2.6 ± 0.1 to 3.1 ± 0.2 mmol/L, P = 0.02), and symptom and plasma metanephrine responses to hypoglycemia were higher (median area under curve for symptoms, 580 [interquartile range {IQR} 420-780] vs. 710 [460-1,260], P = 0.02; metanephrine, 2,412 [-3,026 to 7,279] vs. 5,180 [-771 to 11,513], P = 0.01). Glycemic threshold for deterioration of cognitive function measured by four-choice reaction time was unchanged, while the color-word Stroop test showed a degree of adaptation. CONCLUSIONS: Even in long-standing T1D, IAH and defective counterregulation may be improved by a clinical strategy aimed at hypoglycemia avoidance.

In vivo cardiac glucose metabolism in the high-fat fed mouse: comparison of euglycemic-hyperinsulinemic clamp derived measures of glucose uptake with a dynamic metabolomic flux profiling approach

Rationale Cardiac metabolism is thought to be altered in insulin resistance and type 2 diabetes (T2D). Our understanding of the regulation of cardiac substrate metabolism and insulin sensitivity has largely been derived from ex vivo preparations which are not subject to the same metabolic regulation as in the intact heart in vivo. Studies are therefore required to examine in vivo cardiac glucose metabolism under physiologically relevant conditions. Objective To determine the temporal pattern of the development of cardiac insulin resistance and to compare with dynamic approaches to interrogate cardiac glucose and intermediary metabolism in vivo. Methods and results Studies were conducted to determine the evolution of cardiac insulin resistance in C57Bl/6 mice fed a high-fat diet (HFD) for between 1 and 16 weeks. Dynamic in vivo cardiac glucose metabolism was determined following oral administration of [U-¹³C] glucose. Hearts were collected after 15 and 60 min and flux profiling was determined by measuring ¹³C mass isotopomers in glycolytic and tricarboxylic acid (TCA) cycle intermediates. Cardiac insulin resistance, determined by euglycemic-hyperinsulinemic clamp, was evident after 3 weeks of HFD. Despite the presence of insulin resistance, in vivo cardiac glucose metabolism following oral glucose administration was not compromised in HFD mice. This contrasts our recent findings in skeletal muscle, where TCA cycle activity was reduced in mice fed a HFD. Similar to our report in muscle, glucose derived pyruvate entry into the TCA cycle in the heart was almost exclusively via pyruvate dehydrogenase, with pyruvate carboxylase mediated anaplerosis being negligible after oral glucose administration. Conclusions Under experimental conditions which closely mimic the postprandial state, the insulin resistant mouse heart retains the ability to stimulate glucose metabolism.

Population and individual-scale responses to patch size, isolation and quality in the hazel dormouse

Patch size, isolation and quality are key factors influencing species persistence in fragmented landscapes. However, we still lack a detailed understanding of how these variables exert their effects on populations inhabiting fragmented landscapes. At which ecological scale do they have an effect (e.g., individuals versus populations) and, on which demographic parameters? Answering these questions will identify the mechanisms that underlie population turnover rather than solely predicting it based on proxies (e.g., presence/absence data). We report the results of a large-scale, three-year study focused on the relative effects of patch size, isolation and quality on individuals and populations of an arboreal rodent, the hazel dormouse (Muscardinus avellanarius). We examined 30 sites nested within three landscapes characterized by contrasting levels of habitat amount and habitat quality (food resources). We quantified the effects of patch size and quality on the response of individuals (survival and litter size) and populations (density and colonization/extinction dynamics). We identified demographic mechanisms which led to population turnover. Habitat quality positively affected survival (not litter size) and population density (measured through an index). We infer that the decline in survival due to patch quality reduced patch recolonization rather than increasing extinction, while extinction was mainly affected by patch size. Our findings suggest that the effect of patch quality on individual and population parameters was constrained by the physical structure of the surrounding landscapes. At the same time, our results highlight the importance of preserving habitat quality to help the persistence of entire systems of patches.