On the Pharyngeal Feeding Filter of the Salp Pegea confoederata (Tunicata: Thaliacea)

In Pegea, scanning electron microscopy of what appear to be the least damaged portions of the filter shows that it has a regular rectangular mesh consisting of thick (100 nm) fibres at right angles to thinner (50 nm) fibres. The rectangular pores of the filter are around 3.3 × 0.57 μm. These measured values from filters that have suffered shrinkage (to an uncertain degree) during preparation are considered to indicate that the actual pore size in life is some 4.0 × 0.7 μm. The mucous-net feeding filter of salps differs from that of other tunicates since flow through it results from muscular activity. Calculations based on the estimated pore size and filtering rate suggest that during part of the filtering cycle, the pressure drop across the filter is considerably greater than that across other tunicate mucous-net filters.     

Electrochemically directed synthesis of oligonucleotides for DNA microarray fabrication

We demonstrate a new method for making oligonucleotide microarrays by synthesis in situ. The method uses conventional DNA synthesis chemistry with an electrochemical deblocking step. Acid is delivered to specific regions on a glass slide, thus allowing nucleotide addition only at chosen sites. The acid is produced by electrochemical oxidation controlled by an array of independent microelectrodes. Deblocking is complete in a few seconds, when competing side-product reactions are minimal. We demonstrate the successful synthesis of 17mers and discrimination of single base pair mismatched hybrids. Features generated in this study are 40 μm wide, with sharply defined edges. The synthetic technique may be applicable to fabrication of other molecular arrays.     

The C-value enigma in plants and animals: a review of parallels and an appeal for partnership

• Aims Plants and animals represent the first two kingdomsrecognized, and remain the two best-studied groups in termsof nuclear DNA content variation. Unfortunately, the traditionalchasm between botanists and zoologists has done much to preventan integrated approach to resolving the C-value enigma, thelong-standing puzzle surrounding the evolution of genome size.This grand division is both unnecessary and counterproductive,and the present review aims to illustrate the numerous linksbetween the patterns and processes found in plants and animalsso that a stronger unity can be developed in the future. • Scope This review discusses the numerous parallels thatexist in genome size evolution between plants and animals, including(i) the construction of large databases, (ii) the patterns ofDNA content variation among taxa, (iii) the cytological, morphological,physiological and evolutionary impacts of genome size, (iv)the mechanisms by which genomes change in size, and (v) thedevelopment of new methodologies for estimating DNA contents. • Conclusions The fundamental questions of the C-valueenigma clearly transcend taxonomic boundaries, and increasedcommunication is therefore urged among those who study genomesize evolution, whether in plants, animals or other organisms.     

The effects of time,space and spectrum on auditory grouping in túngara frogs

Male túngara frogs (Physalaemus pustulosus) produce complex calls consisting of two components, a ~350 ms FM sweep called the “whine” followed by up to seven ~40 ms harmonic bursts called “chucks”. In order to choose and locate a calling male, females attending to choruses must group call components into auditory streams to correctly assign calls to their sources. Previously we showed that spatial cues play a limited role in grouping: calls with normal spectra and temporal structure are grouped over wide angular separations (≤135°). In this study we again use phonotaxis to first test whether an alternative cue, the sequence of call components, plays a role in auditory grouping and second, whether grouping is mediated by peripheral or central mechanisms. We found that while grouping is not limited to the natural call sequence, it does vary with the relative onset times of the two calls. To test whether overlapping stimulation in the periphery is required for grouping, the whine and chuck were filtered to restrict their spectra to the sensitivity ranges of the amphibian and basilar papillae, respectively. For these dichotic-like stimuli, grouping still occurred (albeit only to 45° separation), suggesting that stream formation is mediated by central mechanisms.     

Skeletal muscle glycogen synthase subcellular localization: effects of insulin and PPAR-alpha agonist (K-111) administration in rhesus monkeys

Insulin covalently and allosterically regulates glycogen synthase (GS) and may also cause the translocation of GS from glycogen-poor to glycogen-rich locations. We examined the possible role of subcellular localization of GS and glycogen in insulin activation of GS in skeletal muscle of six obese monkeys and determined whether 1) insulin stimulation during a hyperinsulinemic euglycemic clamp and/or peroxisome proliferator-activated receptor (PPAR)-alpha agonist treatment (K-111, 3 mg.kg(-1).day(-1); Kowa) induced translocation of GS and 2) translocation of GS was associated with insulin activation of GS. GS and glycogen were present in all fractions obtained by differential centrifugation, except for the cytosolic fraction, under both basal and insulin-stimulated conditions. We found no evidence for translocation of GS by insulin. GS total (GST) activity was strongly associated with glycogen content (r = 0.70, P < 0.001). Six weeks of treatment with K-111 increased GST activity in all fractions, except the cytosolic fraction, and mean GST activity, GS independent activity, and glycogen content were significantly higher in the insulin-stimulated samples compared with basal samples, effects not seen with vehicle. The increase in GST activity was strongly related to the increase in glycogen content during the hyperinsulinemic euglycemic clamp after K-111 administration (r = 0.74, P < 0.001). Neither GS protein expression nor GS gene expression was affected by insulin or by K-111 treatment. We conclude that 1) in vivo insulin does not cause translocation of GS from a glycogen-poor to a glycogen-rich location in primate skeletal muscle and 2) the mechanism of action of K-111 to improve insulin sensitivity includes an increase in GST activity without an increase in GS gene or protein expression.     

Screening for bacterial-fungal associations in a south-eastern US salt marsh using pre-established fungal monocultures

Both bacteria and fungi play critical roles in decomposition processes in many natural environments, yet only rarely have they been studied as an integrated community. We examined whether physical associations exist between individual bacterial and fungal species that co-occur on decaying smooth cordgrass, Spartina alterniflora, in a south-eastern US salt marsh. Fungal-pervaded decaying Spartina was used as "bait" for potential bacterial associates. The bundles (infiltrated with one of three dominant fungal members of the decomposer assemblage, or an autoclaved control) were placed in a salt marsh and collected biweekly for 6 weeks during the first experiment (late summer 2002), and weekly for 3 weeks during the second experiment (early summer 2003). Terminal-restriction fragment length polymorphism (T-RFLP) analysis of 16S rRNA genes was used to track colonization by bacterial taxa in association with the established fungal species. T-RFLP analysis of 18S-to-28S internal transcribed spacer (ITS) regions was used to monitor changes in fungal communities once bundles had been placed in the field. Results from both years were nearly identical, and showed that invasion by fungi other than the bait species was slow, resulting in a virtual fungal monoculture for several weeks into the experiments. Surprisingly, bacterial communities were unaffected by the identity of the fungal bait. Regardless of the fungal species, and even in the absence of prior fungal colonization, bacterial 16S rRNA profiles were remarkably similar. These results suggest that few species-specific associations, either positive or negative, exist between bacterial and fungal members of the Spartina decomposer community during initial colonization.     

Seasonal patterns in soil surface CO<Subscript>2</Subscript> flux under snow cover in 50 and 300 year old subalpine forests

Soil CO₂ flux can contribute as much as 60–80% of total ecosystem respiration in forests. Although considerable research has focused on quantifying this flux during the growing season, comparatively little effort has focused on non-growing season fluxes. We measured soil CO₂ efflux through snow in 50 and ~300 year old subalpine forest stands near Fraser CO. Our objectives were to quantify seasonal patterns in wintertime soil CO₂ flux; determine if differences in soil CO₂ flux between the two forest ages during the growing season persist during winter; and to quantify the sample size necessary to discern treatment differences. Soil CO₂ flux during the 2002–2003 and 2003–2004 snow season averaged 0.31 and 0.35 μmols m⁻² s⁻¹ for the young and old forests respectively; similar to the relative difference observed during summer. There was a significant seasonal pattern of soil CO₂ flux during the winter with fluxes averaging 0.22 μmols m⁻² s⁻¹ in December and January and increasing to an average of 0.61 μmols m⁻² s⁻¹ in May. Within-plot variability for measurements used in calculating flux was low. The coefficients of variation (CV) for CO₂ concentration, snowpack density, and snow depth were 17, 8 and 14%, respectively, yielding a CV for flux measurements within-plot of 29%. A within plot CV of 29% requires 8 sub-samples per plot to estimate the mean flux with a standard error of ±10% of the mean. Variability in CO₂ flux estimates among plots (size = 400 m²) was similar to that within plot and was also low (CV = ~28%). With a CV of 28% among plots, ten plots per treatment would have a 50% probability of detecting a 25% difference in treatment means for α = 0.05.