Tracking transmitter-gated P2X cation channel activation in vitro and in vivo

We present a noninvasive approach to track activation of ATP-gated P2X receptors and potentially other transmitter-gated cation channels that show calcium fluxes. We genetically engineered rat P2X receptors to carry calcium sensors near the channel pore and tested this as a reporter for P2X(2) receptor opening. The method has several advantages over previous attempts to image P2X channel activation by fluorescence resonance energy transfer (FRET): notably, it reports channel opening rather than a conformation change in the receptor protein. Our FRET-based imaging approach can be used as a general method to track, in real time, the location, regional expression variation, mobility and activation of transmitter-gated P2X channels in living neurons in vitro and in vivo. This approach should help to determine when, where and how different receptors are activated during physiological processes.     

Understanding crown shyness from a 3-D perspective

Background and AimsCrown shyness describes the phenomenon whereby tree crowns avoid growing into each other, producing a puzzle-like pattern of complementary tree crowns in the canopy. Previous studies found that tree slenderness plays a role in the development of crown shyness. Attempts to quantify crown shyness have largely been confined to 2-D approaches. This study aimed to expand the current set of metrics for crown shyness by quantifying the characteristic of 3-D surface complementarity between trees displaying crown shyness, using LiDAR-derived tree point clouds. Subsequently, the relationship between crown surface complementarity and slenderness of trees was assessed.MethodsFourteen trees were scanned using a laser scanning device. Individual tree points clouds were extracted semi-automatically and manually corrected where needed. A metric that quantifies the surface complementarity (S_c) of a pair of protein molecules is applied to point clouds of pairs of adjacent trees. Then 3-D tree crown surfaces were generated from point clouds by computing their α shapes.Key ResultsTree pairs that were visually determined to have overlapping crowns scored significantly lower S_c values than pairs that did not overlap (n = 14, P < 0.01). Furthermore, average slenderness of pairs of trees correlated positively with their S_c score (R² = 0.484, P < 0.01), showing agreement with previous studies on crown shyness.ConclusionsThe characteristic of crown surface complementarity present in trees displaying crown shyness was succesfully quantified using a 3-D surface complementarity metric adopted from molecular biology. Crown surface complementarity showed a positive relationship to tree slenderness, similar to other metrics used for measuring crown shyness. The 3-D metric developed in this study revealed how trees adapt the shape of their crowns to those of adjacent trees and how this is linked to the slenderness of the trees.     

Effects of endogenous and exogenous processes on aboveground biomass stocks and dynamics in Andean forests

Tropical forests are paramount in regulating the global carbon cycle due to the storage of large amounts of carbon in their biomass. Using repeat censuses of permanent plots located at 15 sites in the Andes Mountains of northwest Colombia, we evaluate: (1) the relationship between aboveground biomass (AGB) stocks, AGB dynamics (mortality, productivity, and net change), and changes in temperature across a ca. 3000-m elevational gradient (≈?16.1 °C); (2) how AGB mortality and AGB productivity interact to determine net AGB change; and (3) the extent to which either fine-grain (0.04-ha) or coarse-grain (1-ha) processes determine the AGB dynamics of these forests. We did not find a significant relationship between elevation/temperature and biomass stocks. The net AGB sequestered each year by these forests (2.21?±?0.51 Mg ha^?1 year^?1), equivalent to approximately 1.09% of initial AGB, was primarily determined by tree growth. Both forest structural properties and global warming influenced AGB mortality and net change. AGB productivity increases with greater inequality of tree sizes, a pattern characteristic of forest patches recovering from disturbances. Overall, we find that global warming is triggering directional changes in species composition by thermophilization via increased tree mortality of species in the lower portions of their thermal ranges and that the inclusion of small-scale forest structural changes can effectively account for endogenous processes such as changes in forest structure. The inclusion of fine-grain processes in assessments of AGB dynamics could provide additional insights about the effects that ongoing climate change has on the functioning of tropical montane forests.     

Studies on evolutionary and selective properties of hypercycles using a Monte Carlo method

Summary The most relevant properties of hypercycles were previously studied mainly from a theoretical point of view. We have developed a Monte Carlo method simulating hypercyclic organization to obtain information about the dynamics of this prebiotic organization. Nucleation, growth, and selective properties have been tested and the results obtained are in good agreement with those of the theoretical predictions. The influence of hypercyclic organization of the error threshold has also been studied. As a consequence of the emergence of a hypercycle, the value of this threshold decreases. The amount of this decrease depends on the population size. Moreover, for some interval of quality factor values, either the hypercycle organization or an error catastrophe can be produced, depending on the initial conditions. The influence of these phenomena on both the dynamic behavior and evolutionary advantages of the hypercycle, as well as their decisive roles on genome size, are discussed.Presented at the FEBS Symposium on Genome Organization and Evolution, held in Crete, Greece, September 1–5, 1986     

Fractionation of Eucalyptus grandis chips by dilute acid-catalysed steam explosion

Steam explosion of Eucalyptus grandis has been carried out under various pretreatment conditions (200-210 degrees C, 2-5 min) after impregnation of the wood chips with 0.087 and 0.175% (w/w) H2SO4. This study, arranged as a 2(3) factorial design, indicated that pretreatment temperature is the most critical variable affecting the yield of steam-treated fractions. Pretreatment of 0.175% (w/w) H2SO4-impregnated chips at 210 degrees C for 2 min was the best condition for hemicellulose recovery (mostly as xylose) in the water soluble fraction, reaching almost 70% of the corresponding xylose theoretical yield. By contrast, lower pretreatment temperatures of 200 degrees C were enough to yield steam-treated substrates from which a 90% cellulose conversion was obtained in 48 h, using low enzyme loadings of a Celluclast 1.5 1 plus Novozym 188 mixture (Novo Nordisk). Release of water-soluble chromophores was monitored by UV spectroscopy and their concentration increased with pretreatment severity. The yield of alkali-soluble lignin increased at higher levels of acid impregnation and pretreatment temperatures. Thermoanalysis of these lignin fractions indicated a pattern of lignin fragmentation towards greater pretreatment severities but lignin condensation prevailed at the most drastic pretreatment conditions.     

Biochemical markers of vascular calcification in elderly hemodialysis patients

The increased vascular calcification, cardiovascular morbidity, and mortality in chronic kidney disease (CKD) patients has been associated with disturbances in mineral-bone metabolism. In order to determine markers of the vascular calcification frequently observed in these patients, blood samples of elderly male and female hemodialysis CKD patients were used to measure serum levels of: osteoprotegerin (OPG), total soluble receptor activator of nuclear factor-κB ligand (sRANKL), and fetuin-A by enzyme immunoassay; tartrate-resistant acid phosphatase (TRACP-5b), and bone-specific alkaline phosphatase (BAP) by immunoenzymometric assay; osteocalcin (OC) by ELISA; iPTH by immunoradiometric assay; 25(OH)D₃ and 1,25(OH)₂D₃, by I¹²⁵ radioimmunoassay; and calcium and phosphorus by photometric assay. Serum OPG, BAP, iPTH, phosphorus, and OC levels were higher and serum 25(OH)D₃, 1,25(OH)₂D₃, and fetuin-A levels lower in both male and female CKD patients than in their respective controls. Our results indicate that the bone formation and resorption parameters are altered in elderly male and female hemodialysis CKD patients. These changes may lead to vascular calcifications and cardiovascular complications, given that elevated OPG and OC levels and reduced fetuin-A levels are associated with cardiovascular events.     

Canopy tree mortality depends on the proportion of crown exposed to sunlight,but this effect varies with species' wood density

Understanding what drives changes in tree mortality as well as the covariates influencing trees' response is a research priority to predict forest responses to global change. Here, we combined drone photogrammetry and ground-based data to assess the influence of crown exposure to light (relative to total crown area), growth deviations (relative to conspecifics), tree size, and species' wood density (as a surrogate for light-demanding and shade-tolerant life-history strategies) on the mortality of 984 canopy trees in an Amazon terra firme forest. Trees with lower wood density were less prone to die when their proportion of crown was more exposed to sunlight, but this relationship with relative crown exposure weakened and slightly reversed as wood density increased. Trees growing less than their species average had higher mortality, especially when the species' wood density decreased. The role of wood density in determining the survival of canopy trees under varying light conditions indicates differential responses of light-demanding versus shade-tolerant species. Our results highlight the importance of accounting for life-history strategies, via plant functional types, in vegetation dynamic models aiming to predict forest demography under a rapidly changing climate. Abstract in Spanish is available with online material.     

Current status of American bullfrog, <Emphasis Type="Italic">Lithobates catesbeianus</Emphasis>, invasion in Uruguay and exploration of chytrid infection

The American bullfrog Lithobates catesbeianus is an invasive species that can strongly affect native amphibian communities through competition, predation, or introduction of diseases. This frog has invaded multiple areas in South America, for which niche models predict suitable environments across much of the continent. This paper reveals the state of the invasion of this species in Uruguay and its possible relationship with the chytrid pathogenic fungus, Batrachochytrium dendrobatidis. Surveys at invaded sites were conducted from 2007 to 2015, identified two populations undergoing recent range expansion (one of them exponential), two populations that failed to establish, and a new record in an urban area of the capital city, Montevideo. In all the analysed feral populations, chytridiomycosis was found. Our data suggest that the invasion of L. catesbeianus in Uruguay is at an early stage, with very localized populations, which might allow for the implementation of cost-effective management plans, with eradication constituting a plausible option.     

Genetic characterization of 12 heterologous microsatellite markers for the giant tropical tree Cariniana legalis (Lecythidaceae)

Twelve microsatellite loci previously developed in the tropical tree Cariniana estrellensis were genetically characterized in Cariniana legalis. Polymorphisms were assessed in 28 C. legalis individuals found between the Pardo and Mogi-Guaçu River basins in the state of São Paulo, Brazil. Of the 12 loci, 10 were polymorphic and exhibited Mendelian inheritance. The allelic richness at each locus ranged from 2-11, with an average of 7 alleles per locus, and the expected heterozygosity ranged from 0.07-0.88. These loci showed a high probability of paternity exclusion. The characteristics of these heterologous microsatellite markers indicate that they are suitable tools for investigating questions concerning population genetics in C. legalis.