Tropical forest restoration in Costa Rica: the effect of several strategies on litter production, accumulation and decomposition

Tropical forest restoration strategies have the potential to accelerate the recovery of the nutrient cycles in degraded lands. Litter production and its decomposition represent the main transfer of organic material and nutrients into the soil substrate. We evaluated litter production, accumulation on the forest floor, and its decomposition under three restoration strategies: plantation (entire area planted with trees), island (trees planted in patches of three different sizes) and control (natural regeneration) plots. We also compared restoration strategies to young secondary forest (7-9 yr). Restoration treatments were established in 50 x 50m plots in June 2004 at six sites in Southern Costa Rica. Planted tree species included two native timber species (Terminalia amazonia and Vochysia guatemalensis) interplanted with two N fixers (Erythrina poeppigiana and Inga edulis). Litter was collected every 15 days between September 2008 and August 2009 in 12 0.25m2 litter traps distributed within each plot; litter that accumulated on the soil surface was collected at four locations (0.25m2 quadrats) within each plot in February and May 2009. Total litter production in plantation (6.3Mg/ha) and secondary forest (7.3Mg/ha) did not differ, but were greater than in islands (3.5Mg/ha) and control (1.4 Mg/ha). Plantation had greatest accumulation of litter on the soil surface (10.6 Mg/ha) as compared to the other treatments (SF = 7.2; I = 6.7; C = 4.9). Secondary forest was the only treatment with a greater annual production of litter than litter accumulation on the soil surface. Carbon storage in litter was similar between plantation and secondary forest, and significantly greater than the other treatments. No differences were found for carbon concentration and storage in the soil among treatments. There was also high variability in the production and accumulation of litter and carbon among sites. Active restoration treatments accelerated the production of litter and carbon storage in comparison to areas under natural recovery. However, the nutrient cycle has not necessarily been restored under these conditions, as high litter accumulation on the soil surface indicates a low decomposition rate, which slows nutrient return to the soil.     

Testing Moran's theorem in an agroecosystem

The abundance and reproductive effort of populations frequently fluctuate across space and time, a phenomenon known as spatial synchrony. Knowledge of the causes of this behavior underlies the ability to manage species, protect the health of humans and the environment, and increase agricultural sustainability. We used an agroecosystem to test Moran's theorem – spatial synchrony results from environmental entrainment. The controlled conditions of the agroecosystem allowed us to create a highly correlated environment while negating the effects of the alternative hypotheses: dispersal and trophic interactions. Under such conditions, synchrony of fruit production by 4288 trees was high over six years in a 32.5 ha pistachio orchard and occurred at similar temporal frequency as weather patterns demonstrating the Moran effect. The spatial synchrony of fruit production was less than the presumed synchrony of the environment supporting research from microcosms and observational studies showing the Moran effect is degraded by local mechanisms. Indeed even under the homogeneous environment of this system, synchrony declined significantly with distance among trees. We present evidence suggesting that the correlation of the local environment affects intrinsic dynamics to cause these patterns. Our findings demonstrate that the Moran effect is, at minimum, partially responsible for the synchronous fruit production in this system. Agroecosystems are often overlooked in basic ecological research; this experiment provides an example of their comparative advantages for the study of some ecological questions.     

Discovery of selective glucocorticoid receptor modulator MK-5932

A series of partial agonists of the Glucocorticoid Receptor were prepared targeting reduced transactivation activity, while maintaining significant transrepression activity. Incorporation of an ortho-aryl amide produced compounds with the desired in vitro profile. Bioreactors consisting of Suspension cultures of Sf21 cells co expressing a CYP3A4 and NADPH-cytochrome P450 oxireductase were used to prepare the major metabolites of these compounds and revealed that oxidative N-dealkylation provided a pathway for formation of metabolites that were more agonistic than the parent partial agonists. Oxidative N-dealkylation was blocked in a new series of compounds, however oxidation alone was capable of producing full agonist metabolites. Incorporation of an ortho-primary amide and utilization of fluorine to modulate agonism afforded partial agonist MK-5932. Synthesis of the major metabolites of MK-5932 using bioreactor technology revealed that no significant GR-active metabolites were formed. Orally administered MK-5932 displayed anti-inflammatory efficacy in a Rat Oxazolone-induced chronic dermatitis model, while sparing plasma insulin.     

DIETS OF COASTAL BOTTLENOSE DOLPHINS FROM THE U. S. MID-ATLANTIC COAST DIFFER BY HABITAT

We recorded 31 species in the stomachs of 146 coastal bottlenose dolphins ( Tursiops truncatus ) from North Carolina, U. S. A. Sciaenid fishes were the most common prey (frequency of occurrence = 95%). By mass, Atlantic croaker ( Micropogonias undulatus ) dominated the diet of dolphins that stranded inside estuaries, whereas weakfish ( Cynosicon regalis ) was most important for dolphins in the ocean. Inshore squid ( Loligo sp.) was eaten commonly by dolphins in the ocean, but not by those in the estuaries. There was no significant pattern in prey size associated with dolphin demography, but the proportion of the diet represented by croaker was higher for males than for females, and mature dolphins ate more croaker than did juveniles. Dietary differences between dolphins that stranded in the estuaries and those that stranded on ocean beaches support the hypothesis that some members of the population inhabit the ocean primarily while others reside principally in estuaries. The overwhelming majority of prey were soniferous species (75% of numerical abundance), which is consistent with the hypothesis that bottlenose dolphins use passive listening to locate noise-making fishes. However, spatiotemporal patterns in consumption of Sciaenid fishes did not coincide with their spawning, which is when peak sound production is thought to occur.     

N-Glycosylation regulates endothelial lipase-mediated phospholipid hydrolysis in apoE- and apoA-I-containing high density lipoproteins

Endothelial lipase (EL) is a member of the triglyceride lipase gene family with high phospholipase and low triacylglycerol lipase activities and a distinct preference for hydrolyzing phospholipids in HDL. EL has five potential N-glycosylation sites, four of which are glycosylated. The aim of this study was to determine how glycosylation affects the phospholipase activity of EL in physiologically relevant substrates. Site-directed mutants of EL were generated by replacing asparagine (N) 62, 118, 375, and 473 with alanine (A). These glycan-deficient mutants were used to investigate the kinetics of phospholipid hydrolysis in fully characterized preparations of spherical reconstituted high density lipoprotein (rHDL) containing apolipoprotein E2 (apoE2) [(E2)rHDL], apoE3 [(E3)rHDL], apoE4 [(E4)rHDL], or apoA-I [(A-I)rHDL] as the sole apolipoprotein. Wild-type EL hydrolyzed the phospholipids in (A-I)rHDL, (E2)rHDL, (E3)rHDL, and (E4)rHDL to similar extents. The phospholipase activities of EL N118A, EL N375A, and EL N473A were significantly diminished relative to that of wild-type EL, with the greatest reduction being apparent for (E3)rHDL. The phospholipase activity of EL N62A was increased up to 6-fold relative to that of wild-type EL, with the greatest enhancement of activity being observed for (E2)rHDL. These data show that individual N-linked glycans have unique and important effects on the phospholipase activity and substrate specificity of EL.     

Architecture of the VE-cadherin hexamer

Vascular endothelial-cadherin (VE-cadherin) is the major constituent of the adherens junctions of endothelial cells and plays a key role in angiogenesis and vascular permeability. The ectodomains EC1-4 of VE-cadherin are known to form hexamers in solution. To examine the mechanism of homotypic association of VE-cadherin, we have made a 3D reconstruction of the EC1-4 hexamer using electron microscopy and produced a homology model based on the known structure of C-cadherin EC1-5. The hexamer consists of a trimer of dimers with each N-terminal EC1 module making an antiparallel dimeric contact, and the EC4 modules forming extensive trimeric interactions. Each EC1-4 molecule makes a helical curve allowing some torsional flexibility to the edifice. While there is no direct evidence for the existence of hexamers of cadherin at adherens junctions, the model that we have produced provides indirect evidence since it can be used to explain some of the disparate results for adherens junctions. It is in accord with the X-ray and electron microscopy results, which demonstrate that the EC1 dimer is central to homotypic cadherin interaction. It provides an explanation for the force measurements of the interaction between opposing cadherin layers, which have previously been interpreted as resulting from three different interdigitating interactions. It is in accord with observations of native junctions by cryo-electron microscopy. The fact that this hexameric model of VE-cadherin can be used to explain more of the existing data on adherens junctions than any other model alone argues in favour of the existence of the hexamer at the adherens junction. In the context of the cell-cell junction these cis-trimers close to the membrane, and trans-dimers from opposing membranes, would increase the avidity of the bond.     

Relative contributions of leaf area ratio and net assimilation rate to change in growth rate depend on growth temperature: comparative analysis of subantarctic and alpine grasses

The present study shows that the relative contributions of leaf area ratio (LAR) and net assimilation rate (NAR) to variation among species in relative growth rate (RGR) depend on growth temperature. We grew three subantarctic and three alpine Poa species at daytime temperatures of 7, 12 and 17 degrees C, and analysed interspecific and temperature-related variation in RGRs by growth analysis. Variation in NAR accounted for most of the interspecific differences in RGR at low growth temperature, whereas variation in both NAR and LAR contributed strongly to interspecific differences in RGR at high growth temperature. For most species, the increase in RGR from 7 to 12 degrees C was attributable to an increase in LAR, whereas the increase in RGR from 12 to 17 degrees C was attributable to an increase in NAR. There were no differences between native subantarctic and alpine species in the plasticity of growth responses to temperature. However, Poa annua, a species introduced to the subantarctic, showed much greater growth plasticity than other species. There was little difference among species in tolerance of high-temperature extremes.     

An Efficient Fungal RNA-Silencing System Using the DsRed Reporter Gene

In filamentous fungi, RNA silencing is an attractive alternative to disruption experiments for the functional analysis of genes. We adapted the gene encoding the autofluorescent DsRed protein as a reporter to monitor the silencing process in fungal transformants. Using the cephalosporin C producer Acremonium chrysogenum, strains showing a high level of expression of the DsRed gene were constructed, resulting in red fungal colonies. Transfer of a hairpin-expressing vector carrying fragments of the DsRed gene allowed efficient silencing of DsRed expression. Monitoring of this process by Northern hybridization, real-time PCR quantification, and spectrofluorometric measurement of the DsRed protein confirmed that downregulation of gene expression can be observed at different expression levels. The usefulness of the DsRed silencing system was demonstrated by investigating cosilencing of DsRed together with pcbC, encoding the isopenicillin N synthase, an enzyme involved in cephalosporin C biosynthesis. Downregulation of pcbC can be detected easily by a bioassay measuring the antibiotic activity of individual strains. In addition, the presence of the isopenicillin N synthase was investigated by Western blot hybridization. All transformants having a colorless phenotype showed simultaneous downregulation of the pcbC gene, albeit at different levels. The RNA-silencing system presented here should be a powerful genetic tool for strain improvement and genome-wide analysis of this biotechnologically important filamentous fungus.