Identification of a α‐helical molten globule intermediate and structural characterization of β‐cardiotoxin,an all β‐sheet protein isolated from the venom of Ophiophagus hannah (king cobra)

β‐Cardiotoxin is a novel member of the snake venom three‐finger toxin (3FTX) family. This is the first exogenous protein to antagonize β‐adrenergic receptors and thereby causing reduction in heart rates (bradycardia) when administered into animals, unlike the conventional cardiotoxins as reported earlier. 3FTXs are stable all β‐sheet peptides with 60–80 amino acid residues. Here, we describe the three‐dimensional crystal structure of β‐cardiotoxin together with the identification of a molten globule intermediate in the unfolding pathway of this protein. In spite of the overall structural similarity of this protein with conventional cardiotoxins, there are notable differences observed at the loop region and in the charge distribution on the surface, which are known to be critical for cytolytic activity of cardiotoxins. The molten globule intermediate state present in the thermal unfolding pathway of β‐cardiotoxin was however not observed during the chemical denaturation of the protein. Interestingly, circular dichroism (CD) and NMR studies revealed the presence of α‐helical secondary structure in the molten globule intermediate. These results point to substantial conformational plasticity of β‐cardiotoxin, which might aid the protein in responding to the sometimes conflicting demands of structure, stability, and function during its biological lifetime.     

Willingness of rural communities to reforest with native tree species in central Chile

Economic development of rural people is not always feasible along with concomitant forest restoration, especially when meager reforestation incentives are oriented to poor rural people who probably are not willing to plant native species in their small plots of land. Forest restoration incentives have been created by the Chilean government to engage poor rural people in reforestation using native tree species to recover degraded lands. Our objective was to compare the willingness of people from rural communities to plant native species if they had to bear the costs or if the government did, and we related the answers to environmental and socioeconomic variables. Of the 217 respondents 53.9% were interested in planting native trees if subsidies became available. Interest decreased if the respondents had to pay for the cost, but only slightly. The willingness to reforest was significantly greater at lower distance from the community to the nearest native forest for those with lower income level, and was higher when there was use of nontimber forest products or wood by the respondents. However, in spite of the positive disposition to plant native trees, only 23% of the respondents were interested in planting on their own land, which is a requirement to receive the economic incentives. Most respondents were willing to plant in open sites and on degraded hillsides that surround their communities. We conclude that despite monetary incentives, benefits cannot reach most rural inhabitants because of their lack of interest in reforesting their own land.     

Understanding and predicting frost‐induced tropical tree mortality patterns

Extreme climatic and weather events are increasing in frequency and intensity across the world causing episodes of widespread tree mortality in many forested ecosystems. However, we have a limited understanding about which local factors influence tree mortality patterns, restricting our ability to predict tree mortality, especially within topographically complex tropical landscapes with a matrix of mature and secondary forests. We investigated the effects of two major local factors, topography and forest successional type, on climate‐induced tropical tree mortality patterns using an observational and modeling approach. The northernmost Neotropical dry forest endured an unprecedented episode of frost‐induced tree mortality after the historic February 2011 cold wave hit northwestern Mexico. In a moderately hilly landscape covering mature and secondary tropical dry forests, we surveyed 454 sites for the presence or absence of frost‐induced tree mortality. In addition, across forty‐eight 1 ha plots equally split into the two forest types, we examined 6,981 woody plants to estimate a frost‐disturbance severity metric using the density of frost‐killed trees. Elevation is the main factor modulating frost effects regardless of forest type. Higher occurrence probabilities of frost‐induced tree mortality at lowland forests can be explained by the strong influence of elevation on temperature distribution since heavier cold air masses move downhill during advective frosts. Holding elevation constant, the probability of frost‐induced tree mortality in mature forests was twice that of secondary forests but severity showed the opposite pattern, suggesting a cautious use of occurrence probabilities of tree mortality to infer severity of climate‐driven disturbances. Extreme frost events, in addition to altering forest successional pathways and ecosystem services, likely maintain and could ultimately shift latitudinal and altitudinal range margins of Neotropical dry forests.     

Characterization of the polyanion-induced molten globule-like state of cytochrome c

Cytochrome c (cyt c) undergoes a poly(vinylsulphate) (PVS)-induced transition at slightly acidic pH into a molten globule-like state that resembles the effect that negatively charged membrane surfaces have on this protein. In this work, the thermodynamic properties of the molten globule-like state of cyt c in complex with PVS are studied using differential scanning calorimetry, circular dichroism, fluorescence, and absorbance spectroscopy. The temperature-induced transition of the molten globule-like state of cyt c in the complex with PVS is characterized by a significantly lower calorimetric enthalpy than in the "typical" molten globule state of cyt c, i.e. free protein at pH 2.0 in high ionic strength. Moreover, the thermally-denatured state of cyt c in the complex at pH < 6 contains nearly 50% of the native secondary structure. The dependence of the transition temperature on the pH indicates a role for histidine residues in the destabilization of the cyt c structure in the PVS complex and in stabilization of the denatured state with the residual secondary structure. A comparison of the effects of small anions and polyanions demonstrates the importance of cooperativity among the anions in the destabilization of cyt c. Predictably, other hydrophilic flexible polyanions such as heparin, polyglutamate, and polyadenylate also have a destabilizing effect on the structure of cyt c. However, a correlation between the properties of the polyanions and their effect on the protein stability is still unclear.     

Variation in Sexual Expression in Jacaratia mexicana (Caricaceae) in Southern Mexico: Frequency and Relative Seed Performance of Fruit-Producing Males

Dioecy, the segregation of male and female structures among individuals, is widespread in tropical plants, encompassing 10–30 percent of species in some sites. In many cases, interindividual sex separation is not complete, as individual plants, although nominally dioecious, may produce both types of reproductive structures. A common form of this sexual variation is the production of female structures in otherwise male individuals, commonly referred to as fruiting males. Here we report the existence of fruiting males in the dioecious tropical tree Jacaratia mexicana (Caricaceae). We show that fruiting males can constitute up to 45 percent of all males in some populations of a tropical forest in Southern Mexico. In order to determine the functional significance of fruiting males for the breeding system of J. mexicana , we compared the relative performance of male- and female-borne seeds. Our results show that seeds from fruiting males are three times less likely to germinate and survive than seeds from female trees. Based on relative seed fitness data, and sex ratios in natural populations, we estimate that 6–15 percent of the genes contributed by fruiting males to the next generation are transmitted via ovules, meaning that morphological variation in gender is at least partially accompanied by functional gender variation. Finally, our seed fitness estimates for fruiting males suggest that fruiting males will not replace female plants in natural populations.
Abstract in Spanish is available at http://www.blackwell-synergy.com/loi/btp .     

Frugivory by Small Vertebrates Within a Deforested, Dry Tropical Region of Central America

Small vertebrates were inventoried within three habitat types in a degraded dry forest region of Panama. Animals were classified as frugivorous if they were observed foraging on fruit or if fecal samples contained mostly or exclusively seeds. Overall, we found that eight bat species and 21 bird species consumed fruit. The greatest numbers of birds were observed within live fences and bird species richness was greatest within riparian forests. Bat assemblages were not significantly different between habitats. The implication is that ecosystem services such as seed dispersal may still be functional in this landscape.     

Equimolar Mixture of 2,2,2-Trifluoroethanol and 4-Chloro-1-butanol is a Stronger Inducer of Molten Globule State: Isothermal Titration Calorimetric and Spectroscopic Studies

A mixture of 4-chloro-1-butanol and 2,2,2-Trifluoroethanol (TFE) has been used to generate Molten globule (MG) state of structurally homologous but functionally different proteins bovine α-lactalbumin and hen egg-white lysozyme. The thermal denaturation was done using UV–Visible spectroscopy. From UV–Visible profile, thermal transition was not observed beyond a particular concentration. There was an indication of molten globule state in case of α-lactalbumin from circular dichroism experiments. By intrinsic tryptophan fluorescence, acrylamide and potassium iodide quenching, 8-anilino-naphthalene sulfonic acid (ANS) binding and energy transfer studies the presence of molten globule state was confirmed. Quantitative characterization of MG state and determining the binding thermodynamics of ANS to the MG state was done using Isothermal Titration Calorimetry (ITC). Results show that α-lactalbumin exists in MG state at a particular concentration but lysozyme does not show features of MG state.     

Bat assemblage structure in two dry forests of Colombia: Composition, species richness, and relative abundance

We studied the composition, species richness, and relative abundance of bat assemblages in the Colombian dry forests of Chicamocha and Patía. In Chicamocha, 11 bats of the family Phyllostomidae were captured with mist-nets, corresponding to 85–100% of the potential phyllostomids species in the area. Two bats of the family Vespertilionidae were also captured in Chicamocha. In Patía, 12 species were captured with mist-nets, all Phyllostomidae, representing 72–100% of the estimated total number of species in the zone. Minor differences in number of species and composition were detected among sampling periods in Chicamocha. The most common species in this dry forest were Glossophaga longirostris and Sturnira lilium. In Patía, notable differences in the number of species and composition were observed among sampling periods, and the most common species were Artibeus jamaicensis, Carollia perspicillata and Phyllostomus discolor. Arid-zone dwelling bats were absent in Patía and we suggest that this absence may be associated with the isolation of Patía from other northern dry zones of Colombia since Quaternary times. There was also low abundance of bats in Patía, which appears to be related to human disturbance. The most abundant phyllostomid bat species in the two dry forests studied are those that include fruit and/or nectar-pollen from columnar cacti as an important proportion of their diets.