A phylogenomic tree inferred with an inexpensive PCR-generated probe kit resolves higher-level relationships among Neptis butterflies (Nymphalidae: Limenitidinae)

Recent advances in obtaining reduced representation libraries for next-generation sequencing permit phylogenomic analysis of species-rich, recently diverged taxa. In this study, we performed sequence capture with homemade PCR-generated probes to study diversification among closely related species in a large insect genus to examine the utility of this method. We reconstructed the phylogeny of Neptis Fabricius, a large and poorly studied nymphalid butterfly genus distributed throughout the Old World. We inferred relationships among 108 Neptis samples using 89 loci totaling up to 84 519 bp per specimen. Our taxon sample focused on Palearctic, Oriental and Australasian species, but included 8 African species and outgroups from 5 related genera. Maximum likelihood and Bayesian analyses yielded identical trees with full support for almost all nodes. We confirmed that Neptis is not monophyletic because Lasippa heliodore (Fabricius) and Phaedyma amphion (Linnaeus) are nested within the genus, and we redefine species groups for Neptis found outside of Africa. The statistical support of our results demonstrates that the probe set we employed is useful for inferring phylogenetic relationships among Neptis species and likely has great value for intrageneric phylogenetic reconstruction of Lepidoptera. Based on our results, we revise the following two taxa: Neptis heliodore comb. rev. and Neptis amphion comb. rev.     

Regulation of calcium/calmodulin-dependent protein kinase II docking to N-methyl-D-aspartate receptors by calcium/calmodulin and alpha-actinin

Ca(2+) influx through the N-methyl-d-aspartate (NMDA)-type glutamate receptor leads to activation and postsynaptic accumulation of Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) and ultimately to long term potentiation, which is thought to be the physiological correlate of learning and memory. The NMDA receptor also serves as a CaMKII docking site in dendritic spines with high affinity binding sites located on its NR1 and NR2B subunits. We demonstrate that high affinity binding of CaMKII to NR1 requires autophosphorylation of Thr(286). This autophosphorylation reduces the off rate to a level (t(12) = approximately 23 min) that is similar to that observed for dissociation of the T286D mutant CaMKII (t(12) = approximately 30 min) from spines after its glutamate-induced accumulation (Shen, K., Teruel, M. N., Connor, J. H., Shenolikar, S., and Meyer, T. (2000) Nat. Neurosci. 3, 881-886). CaMKII as well as the previously identified NR1 binding partners calmodulin and alpha-actinin bind to the short C-terminal portion of the C0 region of NR1. Like Ca(2+)/calmodulin, autophosphorylated CaMKII competes with alpha-actinin-2 for binding to NR1. We conclude that the NR1 C0 region is a key site for recruiting CaMKII to the postsynaptic site, where it may act in concert with calmodulin to modulate the stimulatory role of alpha-actinin interaction with the NMDA receptor.     

Should I stay or should I go? Modelling dispersal strategies in saproxylic insects based on pheromone capture and radio telemetry: a case study on the threatened hermit beetle <Emphasis Type="Italic">Osmoderma eremita</Emphasis>

To predict how organisms cope with habitat fragmentation we must understand their dispersal biology, which can be notoriously difficult. We used a novel, multi-pronged approach to study dispersal strategies in the endangered saproxylic hermit beetle Osmoderma eremita, exploiting its pheromone system to intercept high numbers of dispersing individuals, which is not possible with other methods. Mark-release-recapture, using unbaited pitfall traps inside oak hollows and pheromone-baited funnel traps suspended from tree branches, was combined with radio telemetry (in females only) to record displacements. Dispersal, modelled as a probability distribution of net displacement, did not differ significantly between sexes (males versus females recaptured), observation methods (females recaptured versus radio-tracked), or sites of first capture (pitfall trap in tree versus pheromone trap – distance from original dispersal point unknown). A model including all observed individuals yielded a mean displacement of 82 m with 1% dispersing > 1 km. Differences in body length were small between individuals captured in pitfall versus pheromone traps, indicating that dispersal is rarely a condition-dependent response in O. eremita. Individuals captured in pheromone traps were consistently lighter, indicating that most dispersal events occur relatively late in life, which agrees with trap catch data. In addition, most (79%) females captured in pheromone traps were mated, showing that females typically mate before leaving their natal tree. Our data show that integrating odour attractants into insect conservation biology provides a means to target dispersing individuals and could greatly improve our knowledge of dispersal biology in threatened species.     

Scale dependency and functional response in moose habitat selection

Habitat selection can be influenced by the distribution of the habitat types in the landscape as well as net gain in visiting patches of resources, causing individual variation in habitat selection. Moreover, the hypothesis of functional response in habitat selection predicts that the degree of selection of a resource depends on its relative availability. We used radio-telemetry data from individual moose on an island off the coast of northern Norway to evaluate whether the selection of habitat types at the landscape scale differed from the choice of habitat types within the home range, and investigated the functional response in habitat selection by relating individual habitat selection to home range characteristics. At the landscape scale, moose selected for habitat types that provided both good forage and cover, with small differences between sex and age groups. At the home range scale, all individuals selected habitat types that were associated with cover and low human impact. Habitat selection was not modified by local moose density, but was related to home range size at both spatial scales. Larger home ranges contained larger proportions of non-preferred habitat types compared to smaller home ranges. At the home range scale, the selection for a habitat type decreased with its relative availability, indicating a functional response in habitat selection. This suggests that habitat selection is modified by home range size, which influences the availability of habitat types and shapes individual habitat selection patterns. Our results support previous suggestions that analyses of habitat or resource selection should follow a multi-scale approach. Both the relative availability of habitat types as well as individual variation in home range size should be accounted for in order to disentangle the complex mechanisms that contribute to shape patterns of resource selection in animals.     

Aquaporins in complex tissues. II. Subcellular distribution in respiratory and glandular tissues of rat

The molecular pathways for fluid transport in pulmonary, oral,and nasal tissues are still unresolved. Here we use immunocytochemistry and immunoelectron microscopy to define the sites of expression of fouraquaporins in the respiratory tract and glandular epithelia, where theyreside in distinct, nonoverlapping sites. Aquaporin-1 (AQP1) is presentin apical and basolateral membranes of bronchial, tracheal, andnasopharyngeal vascular endothelium and fibroblasts. AQP5 is localizedto the apical plasma membrane of type I pneumocytes and the apicalplasma membranes of secretory epithelium in upper airway and salivaryglands. In contrast, AQP3 is present in basal cells of tracheal andnasopharyngeal epithelium and is abundant in basolateral membranes ofsurface epithelial cells of nasal conchus. AQP4 resides in basolateralmembranes of columnar cells of bronchial, tracheal, and nasopharyngealepithelium; in nasal conchus AQP4 is restricted to basolateralmembranes of a subset of intra- and subepithelial glands. These sitesof expression suggest that transalveolar water movement, modulation ofairway surface liquid, air humidification, and generation ofnasopharyngeal secretions involve a coordinated network of aquaporinwater channels.

     

Death, democracy and public ethical choice

The Danish Council of Ethics...believed that the brain-death criterion should not be accepted without public education and debate. Following the introduction of a spectrum of educational and related activites, a Gallup poll found that 98% of the survey population was aware of the debate over brain-vs-heart criteria and that 80% favoured the adoption of a supplemental brain-death standard... This raises the fundamental question of decisionmaking in pluralist democratic societies, of the limits of democratic involvement in such choices, and of the role of bodies like the Danish Council of Ethics... It must be part of the mission of a governmental bioethical body to use its peculiar expertise to teach and to lead -- to build a popular consensus out of confusion. But in doing so, such a Commission will be steering a dangerous course....     

Television viewing and incident cardiovascular disease: prospective associations and mediation analysis in the EPIC Norfolk Study

Background

Although television viewing time is detrimentally associated with intermediate cardiovascular risk factors, the relationship with incident total (i.e. combined fatal and non-fatal) cardiovascular disease (CVD), non-fatal CVD and coronary heart disease is largely unknown. This study examined whether television viewing time is associated with these three outcomes, independently of physical activity energy expenditure and other confounding variables.

Methodology/Principal Findings

A population-based cohort of 12,608 men and women (aged 61.4±9.0), free from stroke, myocardial infarction and cancer at baseline in 1998–2000 were followed up until 2007 (6.9±1.9 years). Participants self-reported education, smoking, alcohol use, antihypertensive, lipid lowering and antidepressant medication, disease history, total energy intake, sleep duration, physical activity and television viewing. BMI, waist circumference, blood pressure, triglycerides, HDL cholesterol and glycated haemoglobin (HbA_1c) were measured by standardized procedures; a clustered metabolic risk score was constructed. Every one hour/day increase in television viewing was associated with an increased hazard for total (HR = 1.06, 95%CI = 1.03–1.08; 2,620 cases), non-fatal CVD (HR = 1.06, 95%CI = 1.03–1.09; 2,134 cases), and coronary heart disease (HR = 1.08, 95%CI = 1.03–1.13; 940 cases), independent of gender, age, education, smoking, alcohol, medication, diabetes status, CVD family history, sleep duration and physical activity energy expenditure. Energy intake, BMI, waist circumference, blood pressure, triglycerides, HDL cholesterol, HbA_1c and the clustered metabolic risk score only partially mediated these associations.

Conclusions

These results indicate that the most prevalent leisure time (sedentary) behaviour, television viewing, independently contributes to increased CVD risk. Recommendations on reducing television viewing time should be considered.     

Human and mouse mitochondrial orthologs of bacterial ClpX

We have determined the cDNA sequence and exon/intron structure of the human CLPX gene encoding a human ortholog of the E. coli ClpX chaperone and protease subunit. The CLPX gene comprises 14 exons and encodes a 633-amino acid-long precursor polypeptide. The polypeptide contains an N-terminal putative mitochondrial transit peptide, and expression of a full-length ClpX cDNA tagged at its C-terminus (Myc-His) shows that the polypeptide is transported into mitochondria. FISH analysis localized the CLPX gene to human Chromosome (Chr) 15q22.1-22.32. This localization was refined by radiation hybrid mapping placing the CLPX gene 4.6 cR distal to D15S159. Murine ClpX cDNA was sequenced, and the mouse Clpx locus was mapped to a position between 31 and 42 cM offset from the centromere on mouse Chr 9. Experimental observations indicate the presence of a pseudogene in the mouse genome and sequence variability between mouse ClpX cDNAs from different strains. Alignment of the human and mouse ClpX amino acid sequences with ClpX sequences from other organisms shows that they display the typical modular organization of domains with one AAA⁺ domain common to a large group of ATPases and several other domains conserved in ClpX orthologs linked by non-conserved sequences. Notably, a C-4 zinc finger type motif is recognized in human and mouse ClpX. This motif of so far unknown function is present only in a subset of the known ClpX sequences. Received: 5 April 2000 / Accepted: 14 June 2000