Effect of different carbohydrate drinks on whole body carbohydrate storage after exhaustive exercise.

Seven untrained male subjects participated in a double-blind, crossover study conducted to determine the efficacy of different carbohydrate drinks in promoting carbohydrate storage in the whole body and skeletal muscle during recovery from exhaustive exercise. The postabsorptive subjects first completed an exercise protocol designed to deplete muscle fibers of glycogen, then consumed 330 ml of one of three carbohydrate drinks (18.5% glucose polymer, 18.5% sucrose, or 12% sucrose; wt/vol) and also received a primed constant infusion of [1-(13)C]glucose for 2 h. Nonoxidative glucose disposal (3.51 +/- 0.28, 18.5% glucose polymer; 2.96 +/- 0.32, 18.5% sucrose; 2.97 +/- 0.16, 12% sucrose; all mmol. kg(-1). h(-1)) and storage of muscle glycogen (5.31 +/- 1.11, 18.5% glucose polymer; 4.07 +/- 1.05, 18.5% sucrose; 3.45 +/- 0.85, 12% sucrose; all mmol. kg wet wt(-1). h(-1); P < 0.05) were greater after consumption of the glucose polymer drink than after either sucrose drink. The results suggest that the consumption of a glucose polymer drink (containing 61 g carbohydrate) promotes a more rapid storage of carbohydrate in the whole body, skeletal muscle in particular, than an isoenergetic sucrose drink.     

Simple and sensitive procedure for the assay of serotonin and catecholamines in brain by high-performance liquid chromatography using fluorescence detection

A simple, rapid and specific method for the determination of serotonin and catecholamines in brain is described. After tissue homogenisation, catecholamines are isolated by adsorption onto alumina and elution with perchloric acid. Serotonin is isolated by extraction into n-heptanol and back-extraction into acid. High-performance liquid chromatography of the acid extracts is performed with a C₁₈ reversed-phase column and simple mobile phases. Detection is by the intrinsic fluorescence of the amines on excitation at 200 nm. Detection limits are 100 pg for norepinephrine, 300 pg for dopamine and 20 pg for serotonin. The results are found to correlate well with a catechol O-methyl transferase radioenzymatic assay for catecholamines and a ninhydrin derivatisation procedure for serotonin.     

Crystallization and preliminary X-ray analysis of phosphoporin from the outer membrane of Escherichia coli.

Phosphoporin is a pore-forming transmembrane protein that spans the outer membrane of Escherichia coli and facilitates the diffusion of phosphates and phosphorylated compounds. Phosphoporin has been crystallized in several different crystal forms, although only one appears to be suitable for X-ray analysis. These crystals, which are hexagonal plates, diffract X-rays to 3 A resolution and belong to the space-group P6(3)22, with unit cell dimensions a = b = 121 A and c = 111 A.     

A Cytotoxic,Co-operative Interaction Between Energy Deprivation and Glutamate Release From System xc? Mediates Aglycemic Neuronal Cell Death

The astrocyte cystine/glutamate antiporter (system x_c⁻) contributes substantially to the excitotoxic neuronal cell death facilitated by glucose deprivation. The purpose of this study was to determine the mechanism by which this occurred. Using pure astrocyte cultures, as well as, mixed cortical cell cultures containing both neurons and astrocytes, we found that neither an enhancement in system x_c⁻ expression nor activity underlies the excitotoxic effects of aglycemia. In addition, using three separate bioassays, we demonstrate no change in the ability of glucose-deprived astrocytes—either cultured alone or with neurons—to remove glutamate from the extracellular space. Instead, we demonstrate that glucose-deprived cultures are 2 to 3 times more sensitive to the killing effects of glutamate or N-methyl-D-aspartate when compared with their glucose-containing controls. Hence, our results are consistent with the weak excitotoxic hypothesis such that a bioenergetic deficiency, which is measureable in our mixed but not astrocyte cultures, allows normally innocuous concentrations of glutamate to become excitotoxic. Adding to the burgeoning literature detailing the contribution of astrocytes to neuronal injury, we conclude that under our experimental paradigm, a cytotoxic, co-operative interaction between energy deprivation and glutamate release from astrocyte system x_c⁻ mediates aglycemic neuronal cell death.     

UniqTag: Content-Derived Unique and Stable Identifiers for Gene Annotation

When working on an ongoing genome sequencing and assembly project, it is rather inconvenient when gene identifiers change from one build of the assembly to the next. The gene labelling system described here, UniqTag, addresses this common challenge. UniqTag assigns a unique identifier to each gene that is a representative k-mer, a string of length k, selected from the sequence of that gene. Unlike serial numbers, these identifiers are stable between different assemblies and annotations of the same data without requiring that previous annotations be lifted over by sequence alignment. We assign UniqTag identifiers to ten builds of the Ensembl human genome spanning eight years to demonstrate this stability. The implementation of UniqTag in Ruby and an R package are available at https://github.com/sjackman/uniqtag sjackman/uniqtag. The R package is also available from CRAN: install.packages ("uniqtag"). Supplementary material and code to reproduce it is available at https://github.com/sjackman/uniqtag-paper.     

Into the light: diurnality has evolved multiple times in geckos

Geckos are the only major lizard group consisting mostly of nocturnal species. Nocturnality is presumed to have evolved early in gecko evolution and geckos possess numerous adaptations to functioning in low light and at low temperatures. However, not all gecko species are nocturnal and most diurnal geckos have their own distinct adaptations to living in warmer, sunlit environments. We reconstructed the evolution of gecko activity patterns using a newly generated time‐calibrated phylogeny. Our results provide the first phylogenetic analysis of temporal activity patterns in geckos and confirm an ancient origin of nocturnality at the root of the gecko tree. We identify multiple transitions to diurnality at a variety of evolutionary time scales and transitions back to nocturnality occur in several predominantly diurnal clades. The scenario presented here will be useful in reinterpreting existing hypotheses of how geckos have adapted to varying thermal and light environments. These results can also inform future research of gecko ecology, physiology, morphology and vision as it relates to changes in temporal activity patterns. © 2015 The Linnean Society of London, Biological Journal of the Linnean Society, 2015, ●●, ●●–●●.     

Secondary structure prediction and determination of proteins--a review

The rapid increase in sequence data in combination with a greater understanding of the forces regulating protein structure has been the impetus for an upsurge in the development of theoretical prediction methods. These methods have afforded protein chemists the ability to identify and quantify the various secondary structures along the protein chain. Concurrently, various physico-chemical techniques have been developed such as nuclear Overhauser enhancement n.m.r. and laser Raman spectroscopy. In addition, traditional methods such as infrared and circular dichroism spectroscopy have been refined. Although both predictive and physico-chemical techniques are limited in the types of secondary structure they are capable of determining, they have provided valuable information with regards to protein folding and topology in the absence of X-ray data, and have formed the basis for the development of improved methods for secondary structure determination. This paper reviews some of the predictive and physico-chemical methods presently used to determine protein secondary structure.     

Soluble Reactive Phosphorus Transport and Retention in Tropical, Rainforest Streams Draining a Volcanic and Geothermally Active Landscape in Costa Rica. : Long-Term Concentration Patterns, Pore Water Environment and Response to ENSO Events

Soluble reactive phosphorus (SRP) transport/retention was determined at four sites in three rainforest streams draining La Selva Biological Station, Costa Rica. La Selva is located at the base of the last remaining intact rainforest transect from 30 m above sea level to 3000 m along the entire Caribbean slope of Central America. Steam SRP levels can be naturally high there due to regional, geothermal groundwater discharged at ambient temperature. Monitoring since 1988 has revealed distinctive long-term differences in background SRP and total P (TP) for three streams in close proximity, and identified the impact of ENSO (El Nino Southern Oscillation) events on SRP-enriched reaches. Mean interannual SRP concentrations (± standard deviation) were 89 ± 53μg/l in the Salto (1988–1996), 21 ± 39μg/l in the Pantano (1988–1998), and 26 ± 35μg/l in the Sabalo (1988–1996). After January, 1997 the separate upland-lowland contributions to discharge and SRP load were determined monthly in the Salto. SRP in Upper Salto was low (19 ± 8μg/l, 1997–2002) until enriched at␣the upland-lowland transition by regional groundwater. Mean SRP concentration in Lower␣Salto (108 ± 104μg/l) was typically highest February–April, the driest months, and lowest July–September, the wettest. SRP concentration was positively correlated to the inverse of discharge in Lower Salto when ENSO data were omitted (1992 and 1998–1999), but not in the Upper Salto, Pantano, or Sabalo. TP was positively correlated to the inverse of discharge in all three streams when ENSO data were omitted. High SRP springs and seeps along the Lower Salto contributed 36% of discharge but 85% of SRP export 1997–2001. Annual SRP flux from the total Salto watershed (1997–2001) averaged 2.9 kg/ha year, but only 0.6 kg/ha year from the Upper Salto. A dye tracer injection showed that pore water environments were distinctly different between Upper and Lower Salto. Upper Salto had high surface water–pore water exchange, high dissolved oxygen, low SRP, and low conductivity similar to surface water, and Lower Salto had low surface water–pore water exchange, low dissolved oxygen, high SRP, and high conductivity reflecting geothermal groundwater influence. SRP export from the Salto was controlled by regional groundwater transfer, which in similar volcanic settings could be a significant P source. However, ENSO events modified the SRP concentration in the Salto suggesting that long-term monitoring is required to understand underlying SRP dynamics and P flux to downstream communities.