Thermoregulatory behaviour explains countergradient variation in the upper thermal limit of a rainforest skink

The vulnerability of a terrestrial ectotherm to high environmental temperatures depends on the animal's thermal physiology and thermoregulatory behaviour. These variables – environment, physiology, and behaviour – interact with each other, complicating assessment of species vulnerability to global warming. We previously uncovered a counterintuitive pattern in rainforest sunskinks Lampropholis coggeri: a negative relationship between their critical thermal maximum (CT_max) and the temperature of their environment. Could this result be explained by a three‐way interaction between environment, physiology, and behaviour? Here we find that sunskink thermal preference is correlated positively with CT_max, but, importantly, skinks from hotter environments prefer lower temperatures than conspecifics from cooler environments. In an acclimation experiment, we find that CT_max is plastic and shifts in alignment with acclimation temperature. We also found heritable variation in this trait in a common garden study, but this variation was small relative to the plastic shifts observed in CT_max. Thus, our previous observation of a negative correlation between field CT_max and temperature is explained, at least in part, by the lizard's thermoregulatory behaviour: lizards from hot environments preferentially choose cool microenvironments, and their physiology acclimates to these cooler experienced temperatures. Our results suggest that behavioural adjustments to the environment can produce countergradient variation in physiological traits. More broadly, our work underscores the importance of interactions between environment, behaviour, and physiology in ectotherms. Understanding these interactions will be crucial in assessing vulnerability to climate change.     

Variability in early post-settlement mortality in intertidal mussels and the role of size at settlement

Early post-settlement mortality is difficult to measure in mobile benthic taxa such as mussels. Field studies typically employ settlement collectors that often contain empty shells of dead individuals, which are usually disregarded. Here, empty shells of pediveligers, i.e. recent settlers that died before beginning to grow as juveniles, were used to assess temporal and spatial patterns of early mortality in mussels, whether size at settlement determined mortality, and whether mortality was density-dependent. This study took place at two intertidal sites in a large harbour in central New Zealand where monthly cohorts of mussels were collected over a 2-year period. Monthly mortality varied substantially, ranging from 0% to 42.6%, but not across the two sites that were 27?km apart. Although there was no density-dependent mortality evident within cohorts, the proportion of pediveligers which were dead on collection was highly positively correlated across sites. Live settlers were on average larger than dead settlers, and the size distribution of dead settlers was shifted toward smaller sizes compared to live settlers. At the extremes, of all settlers with shell length of 260?µm or less, 72% were dead on collection. By contrast, for settlers 310?µm or greater in length, 6% were dead. Together, these results (1) suggest that in this system size at settlement may be an important determinant of very early, peri-metamorphic mortality in mussels, with smaller settlers at higher risk, and (2) demonstrate the value in evaluating dead pediveligers for assessing natural early post-settlement mortality in mussels.     

Design and characterization of MP0250, a tri-specific anti-HGF/anti-VEGF DARPin® drug candidate

MP0250 is a multi-domain drug candidate currently being tested in clinical trials for the treatment of cancer. It comprises one anti-vascular endothelial growth factor-A (VEGF-A), one anti-hepatocyte growth factor (HGF), and two anti-human serum albumin (HSA) DARPin® domains within a single polypeptide chain. While there is first clinical validation of a single-domain DARPin® drug candidate, little is known about DARPin® drug candidates comprising multiple domains. Here, we show that MP0250 can be expressed at 15 g/L in soluble form in E. coli high cell-density fermentation, it is stable in soluble/frozen formulation for 2 years as assessed by reverse phase HPLC, it has picomolar potency in inhibiting VEGF-A and HGF in ELISA and cellular assays, and its domains are simultaneously active as shown by surface plasmon resonance. The inclusion of HSA-binding DARPin® domains leads to a favorable pharmacokinetic profile in mouse and cynomolgus monkey, with terminal half-lives of ～ 30 hours in mouse and ～ 5 days in cynomolgus monkey. MP0250 is thus a highly potent drug candidate that could be particularly useful in oncology. Beyond MP0250, the properties of MP0250 indicate that multi-domain DARPin® proteins can be valuable next-generation drug candidates.     

Role of Polyamines in Apoptosis and Other Recent Advances in Plant Polyamines

Numerous citations in the literature indicate that polyamines are intensively studied in plants. Polyamines are implicated in many functions of the plant cell. Excellent recent reviews cover much of this original research. This article is focused primarily on literature that relates research on the role of polyamines in apoptosis and programmed cell death in both plants and animals. Apoptosis and programmed cell death are considerably better studied in animal systems, and this review demonstrates that the role of polyamines in these processes in plant systems are remarkably congruent with what is known in animal systems. In addition, key recent research reports are reviewed that describe the functional analysis of key polyamine biosynthesis genes in plants in relation to responses to environmental stress signals. Molecular analysis is providing strong evidence for the polyamine biosynthetic pathways to play major roles in ameliorating plant responses to abiotic stresses.     

Phylogeography of the southern skua complex-rapid colonization of the southern hemisphere during a glacial period and reticulate evolution

Whilst we have now a good understanding how past glaciation influenced species at the northern hemisphere, our knowledge of patterns and modes of speciation is far more limited for the southern hemisphere. We provide mtDNA based data on the phylogeography of a circumpolar distributed southern hemisphere seabird group-the southern skua complex (Catharacta spp.). Diversification of southern skuas dates between 210,000 yBP and 150,000 yBP and coincides with a glacial spanning 230,000-140,000 yBP. Skuas most likely first inhabited the Antarctic continent, in the course of global cooling and increasing glaciation spread to the sub-antarctic islands and Tristan da Cunha and finally colonized Patagonia and the Falkland Islands at the glacial maximum. Despite significant differences between taxa most populations still exchange genes with neighboring populations of other taxa and speciation is incomplete.     

Dating of divergences within the Rattus genus phylogeny using whole mitochondrial genomes

The timing and order of divergences within the genus Rattus have, to date, been quite speculative. In order to address these important issues we sequenced six new whole mitochondrial genomes from wild-caught specimens from four species, Rattus exulans, Rattus praetor, Rattus rattus and Rattus tanezumi. The only rat whole mitochondrial genomes available previously were all from Rattus norvegicus specimens. Our phylogenetic and dating analyses place the deepest divergence within Rattus at approximately 3.5 million years ago (Mya). This divergence separates the New Guinean endemic R. praetor lineage from the Asian lineages. Within the Asian/Island Southeast Asian clade R. norvegicus diverged earliest at approximately 2.9Mya. R. exulans and the ancestor of the sister species R. rattus and R. tanezumi subsequently diverged at approximately 2.2Mya, with R. rattus and R. tanezumi separating as recently as approximately 0.4Mya. Our results give both a better resolved species divergence order and diversification dates within Rattus than previous studies.     

Hosts are more important than destinations: What genetic variation in Microctonus aethiopoides (Hymenoptera: Braconidae) means for foreign exploration for natural enemies

Nucleotide sequence data were generated from the gene regions COI, 16S, and arginine kinase to assess genetic variation within the Palearctic parasitoid, Microctonus aethiopoides, reared from Sitona discoideus, S. hispidulus, and Hypera postica collected from two proximate locations in Mediterranean France. Partitioned Bayesian phylogenetic analyses of the molecular data provided strong support for the presence of at least two M. aethiopoides biotypes, one associated with Hypera species and the other with Sitona species. These new results combined with previously published data from 14 countries show that M. aethiopoides genetic variation is much more strongly correlated with host taxon than with sampling location. This contrasts with earlier perceptions that M. aethiopoides exhibits significant geographic variation, and helps to explain the widely varying biological control outcomes that have been obtained following the introductions of M. aethiopoides to Australia, New Zealand, and North America. The results strongly suggest that success rates and environmental safety in biological control would both be improved by ensuring that parasitoids collected in the native range are reared from the same host species as the one being targeted for control in the region of introduction. The results also provided insights both on the evolution of M. aethiopoides' host range, and on its evolutionary transition between solitary and gregarious larval development.     

The triple-120 meter shuttle test: a sport-specific test for assessing anaerobic endurance fitness in rugby league players

The aim of this study was to design a simple field test to measure the anaerobic endurance fitness of rugby league players, which is an important fitness quality in the game of rugby league. Twelve amateur football players with a mean (+/-SD) age of 21.5 years (+/-2.2) volunteered to participate in the study. The subjects completed 1 trial of the Wingate 60-second (W60) cycle test and 2 trials of the new Triple-120 meter shuttle (T120S) test. All trials were completed 4 days apart. The validity of the T120S was determined by comparing physiological responses (heart rate and blood lactate) and rating of perceived exertion to the all-out W60 cycle test. The results indicate there is a significant relationship between maximum heart rate (r = 0.63 and 0.71) for the 2 trials of the T120S and the W60 cycle test. There was no significant relationship between the 2 trials and the W60 cycle test for post 3 minute lactate (r = 0.112 and 0.101) and rating of perceived exertion (r = 0.94 and 0.161). However, the T120S test elicited greater mean values for these measures than the W60 cycle test. The results indicate that the T120S is a valid test of anaerobic endurance and represents a sports specific test of this quality that may provide useful information for players and coaches involved in the sport of rugby league.     

Development of DNA aptamers for cytochemical detection of acetylcholine

This report describes a novel approach to the detection of acetylcholine using DNA aptamers. Aptamers were developed by eight rounds of acetylcholine affinity column chromatography and polymerase chain reaction (PCR) amplification. Sequences from rounds 5 and 8 were screened by colorimetric enzyme-based microtiter plate assays and found to bind acetylcholine and related compounds, but not unrelated compounds. One of the highest affinity aptamers, designated ACh 6R, was further tested in aptamer-peroxidase and aptamer-fluorescence staining protocols. Using Neuro-2a murine neuroblastoma cells induced to differentiate in the presence of 1 μM all-trans-retinoic acid for 5–7 d, ACh 6R detected cholinergic cells by both the peroxidase and fluorescence methods. Unrelated DNA aptamers did not stain the cells using either method. Fixation with cold 2% paraformaldehyde was compared to cold alkaline allyl alcohol plus glutaraldehyde for immobilization of acetylcholine in situ and appeared to enable detection of greater numbers of cholinergic cells, although differences in levels of differentiation may have been a factor as well. Acetylcholine generally appeared to be distributed throughout the differentiated Neuro-2a cell bodies. However, in some cells, punctate staining along neurite outgrowths and near the termini of cellular processes suggested detection of acetylcholine in discrete vesicles.