Salt-induced enhancement of antifreeze protein activity: a salting-out effect

Antifreeze proteins are a structurally diverse group of proteins characterized by their unique ability to cause a separation of the melting- and growth-temperatures of ice. These proteins have evolved independently in different kinds of cold-adapted ectothermic animals, including insects and fish, where they protect against lethal freezing of the body fluids. There is a great variability in the capacity of different kinds of antifreeze proteins to evoke the antifreeze effect, but the basis of these differences is not well understood. This study reports on salt-induced enhancement of the antifreeze activity of an antifreeze protein from the longhorn beetle Rhagium inquisitor (L.). The results imply that antifreeze activity is predetermined by a steady-state distribution of the antifreeze protein between the solution and the ice surface region. The observed salt-induced enhancement of the antifreeze activity compares qualitatively and quantitatively with salt-induced lowering of protein solubility. Thus, salts apparently enhance antifreeze activity by evoking a solubility-induced shift in the distribution pattern of the antifreeze proteins in favour of the ice. These results indicate that the solubility of antifreeze proteins in the solution surrounding the ice crystal is a fundamental physiochemical property in relation to their antifreeze potency.     

Can minimum convex polygon home ranges be used to draw biologically meaningful conclusions?

Many conclusions about mammalian ranging behaviour have been drawn based on minimum convex polygon (MCP) estimates of home range size, although several studies have revealed its unpredictable nature compared to that of the kernel density estimator. We investigated to what extent the choice of home range estimator affected the biological interpretation in comparative studies. We found no discrepancy when the question asked covered a wide range of taxa, as real and very large differences in range size were likely to have masked smaller differences due to the choice of home range estimator. However, when the question asked concerned within-species characteristics, the choice of home range estimator explained as much of the variation in range size as did the ecological variable in question. The implications for macro-ecological and intraspecific studies are discussed.     

Aquaporins in complex tissues: distribution of aquaporins 1-5 in human and rat eye

Multiple physiological fluid movements areinvolved in vision. Here we define the cellular and subcellular sitesof aquaporin (AQP) water transport proteins in human and rat eyes byimmunoblotting, high-resolution immunocytochemistry, and immunoelectronmicroscopy. AQP3 is abundant in bulbar conjunctival epithelium andglands but is only weakly present in corneal epithelium. In contrast, AQP5 is prominent in corneal epithelium and apical membranes of lacrimal acini. AQP1 is heavily expressed in scleral fibroblasts, corneal endothelium and keratocytes, and endothelium covering thetrabecular meshwork and Schlemm's canal. Although AQP1 is plentiful inciliary nonpigmented epithelium, it is not present in ciliary pigmentedepithelium. Posterior and anterior epithelium of the iris and anteriorlens epithelium also contain significant amounts of AQP1, but AQP0(major intrinsic protein of the lens) is expressed in lens fiber cells.Retinal Müller cells and astrocytes exhibit notableconcentrations of AQP4, whereas neurons and retinal pigment epitheliumdo not display aquaporin immunolabeling. These studies demonstrateselective expression of AQP1, AQP3, AQP4, and AQP5 in distinct ocularepithelia, predicting specific roles for each in the complex networkthrough which water movements occur in the eye.

     

Epac1 and cAMP-dependent protein kinase holoenzyme have similar cAMP affinity, but their cAMP domains have distinct structural features and cyclic nucleotide recognition

The cAMP-dependent protein kinase (PKA I and II) and the cAMP-stimulated GDP exchange factors (Epac1 and -2) are major cAMP effectors. The cAMP affinity of the PKA holoenzyme has not been determined previously. We found that cAMP bound to PKA I with a K(d) value (2.9 microM) similar to that of Epac1. In contrast, the free regulatory subunit of PKA type I (RI) had K(d) values in the low nanomolar range. The cAMP sites of RI therefore appear engineered to respond to physiological cAMP concentrations only when in the holoenzyme form, whereas Epac can respond in its free form. Epac is phylogenetically younger than PKA, and its functional cAMP site has presumably evolved from site B of PKA. A striking feature is the replacement of a conserved Glu in PKA by Gln (Epac1) or Lys (Epac2). We found that such a switch (E326Q) in site B of human RIalpha led to a 280-fold decreased cAMP affinity. A similar single switch early in Epac evolution could therefore have decreased the high cAMP affinity of the free regulatory subunit sufficiently to allow Epac to respond to physiologically relevant cAMP levels. Molecular dynamics simulations and cAMP analog mapping indicated that the E326Q switch led to flipping of Tyr-373, which normally stacks with the adenine ring of cAMP. Combined molecular dynamics simulation, GRID analysis, and cAMP analog mapping of wild-type and mutated BI and Epac1 revealed additional differences, independent of the Glu/Gln switch, between the binding sites, regarding space (roominess), hydrophobicity/polarity, and side chain flexibility. This helped explain the specificity of current cAMP analogs and, more importantly, lays a foundation for the generation of even more discriminative analogs.     

Is hunting mortality additive or compensatory to natural mortality? Effects of experimental harvest on the survival and cause-specific mortality of willow ptarmigan

1. The effects of harvest on the annual and seasonal survival of willow ptarmigan Lagopus lagopus L. were tested in a large-scale harvest experiment. Management units were randomly assigned to one of three experimental treatments: 0%, 15% or 30% harvest. Seasonal quotas were based on the experimental treatment and estimates of bird density before the hunting season. Survival rates and hazard functions for radio-marked ptarmigan were then estimated under the competing risks of harvest and natural mortality. 2. The partially compensatory mortality hypothesis was supported: annual survival of ptarmigan was 0·54 ± 0·08 SE under 0% harvest, 0·47 ± 0·06 under 15% harvest, and was reduced to 0·30 ± 0·05 under 30% harvest. Harvest mortality increased linearly from 0·08 ± 0·05, 0·27 ± 0·05 and 0·42 ± 0·06 from 0% to 30% harvest, whereas natural mortality was 0·38 ± 0·08, 0·25 ± 0·05 and 0·28 ± 0·06 under the same treatments. 3. Realized risk of harvest mortality was 0·08-0·12 points higher than our set harvest treatments of 0-30% because birds were exposed to risk if they moved out of protected areas. The superadditive hypothesis was supported because birds in the 30% harvest treatment had higher natural mortality during winter after the hunting season. 4. Natural mortality was mainly because of raptor predation, with two seasonal peaks in fall and spring. Natural and harvest mortality coincided during early autumn with little potential for compensation during winter months. Peak risk of harvest mortality was 5× higher than natural mortality. Low natural mortality during winter suggests that most late season harvest would be additive mortality. 5. Environmental correlates of natural mortality of ptarmigan included seasonal changes in snow cover, onset of juvenile dispersal, and periods of territorial activity. Natural mortality of ptarmigan was highest during autumn movements and nesting by gyrfalcons Falco rusticolus L. Mortality was low when gyrfalcons had departed for coastal wintering sites, and during summer when ptarmigan were attending nests and broods. 6. Our experimental results have important implications for harvest management of upland gamebirds. Seasonal quotas based on proportional harvest were effective and should be set at ≤ 15% of August populations for regional management plans. Under threshold harvest of a reproductive surplus, 15% harvest would be sustainable at productivity rates ≥ 2·5 young per pair. Impacts of winter harvest could be minimized by closing the hunting season in early November or by reducing late season quotas.     

Do ice nucleating lipoproteins protect frozen insects against toxic chemical agents?

As the body fluid of freeze-tolerant organisms freezes, solutes become concentrated in the gradually smaller unfrozen fluid fraction, and dissolved trace metals may reach toxic levels. A dialysis technique was used to investigate the metal binding capacity of the low density fraction of the hemolymph from the freeze tolerant beetle Phyto depressus. The low density fraction, assumed to contain the ice nucleating lipoproteins, showed approximately 100 times greater capacity to bind metals (Cd ²⁺, Cu ²⁺ and Zn ²⁺) than the proteins albumin, hemoglobin and similar to metallothionein. The high metal binding capacity in the low density fraction raises the question if the ice nucleating lipoproteins might assist in detoxification of potentially toxic concentrations of metals that may occur when a large fraction of the bodyfluids of freeze tolerant insects freeze. This hypotheis is consistent with the fact that the lipoprotein ice nucleators are present in far greater amounts than required for ice nucleation, and also with the fact that the lipoprotein ice nucleators have a remarkably high content of amino acids with negatively charged residues that may act as metal binding sites.     

The risks of learning: confounding detection and demographic trend when using count‐based indices for population monitoring

Theory recognizes that a treatment of the detection process is required to avoid producing biased estimates of population rate of change. Still, one of three monitoring programmes on animal or plant populations is focused on simply counting individuals or other fixed visible structures, such as natal dens, nests, tree cavities. This type of monitoring design poses concerns about the possibility to respect the assumption of constant detection, as the information acquired in a given year about the spatial distribution of reproductive sites can provide a higher chance to detect the species in subsequent years. We developed an individual‐based simulation model, which evaluates how the accumulation of knowledge about the spatial distribution of a population process can affect the accuracy of population growth rate estimates, when using simple count‐based indices. Then, we assessed the relative importance of each parameter in affecting monitoring performance. We also present the case of wolverines (Gulo gulo) in southern Scandinavia as an example of a monitoring system with an intrinsic tendency to accumulate knowledge and increase detectability. When the occupation of a nest or den is temporally autocorrelated, the monitoring system is prone to increase its knowledge with time. This happens also when there is no intensification in monitoring effort and no change in the monitoring conditions. Such accumulated knowledge is likely to increase detection probability with time and can produce severe bias in the estimation of the rate and direction of population change over time. We recommend that a systematic sampling of the population process under study and an explicit treatment of the underlying detection process should be implemented whenever economic and logistical constraints permit, as failure to include detection probability in the estimation of population growth rate can lead to serious bias and severe consequences for management and conservation.     

Habitat selection as a hierarchical spatial process: the green woodpecker at the northern edge of its distribution range

Habitat selection can be envisaged as a hierarchical spatial process, from choice of home range to choice of dietary item. The green woodpecker (Picus viridis) is described as being closely bound to cultivated land and deciduous forests, mainly due to its summer diet composed of ants (Hymenoptera: Formicidae) found on meadows and pastures. To explore possible responses of this woodpecker to recent changes in land use practice, we studied home ranges, feeding habitats and food selection of a marginal population (four radio-marked males and five females) in a 30,000-ha conifer-dominated landscape at the northern edge of its distribution range in south-central Scandinavia. We asked: (1) Is the green woodpecker confined to areas with cultivated land and deciduous forest? (2) If so, are important food items (ants) particularly abundant or exclusively found there? (3) Can clearcuts and young plantations substitute for cultivated land as feeding habitat? Home ranges (mean=100 ha) were invariably confined to the parts of the landscape that contained cultivated land (<1% of the total area). In summer, birds preferred to feed in cultivated land, presumably due to a higher overall biomass of ants compared to forest habitats. They avoided clearcuts, but preyed extensively upon soil-dwelling ants in young conifer stands (16–30 years old). We failed to find preferences for particular ant groups (Lasius niger and L. flavus) associated with cultivated land. The principal summer food was Serviformica, an ant group that was equally abundant in cultivated land and forest habitat. A positive correlation between ant body mass and a preference index suggests that the birds selected the larger ant species independent of habitat type. In winter, birds fed exclusively on mound-building Formica rufa-ants in closed-canopy, older forest stands. Our results indicate that the green woodpecker successfully utilizes young conifer plantations as feeding habitat. At a larger scale, we hypothesize that green woodpecker populations fail to establish in managed forest tracts, not because of food shortage, but because the landscapes lack cultivated land serving as a key stimulus encouraging individuals to settle.