Riparian zone as a main determinant of the structure of lizard assemblages in upland Amazonian forests

The use of lizards as model organisms in ecological studies is based on their success in occupying a great diversity of habitats, and some species are closely tied to the environment, which is disadvantaged by the legislation of several countries concerning land use. Our aim was to relate lizard species distribution patterns in rainforest environments to variation in environmental gradients, and provide ecologically based metrics for establishing buffer zones around streams. Lizards were sampled three times in 41 standardised transects near Manaus, Brazil, only in dry season, with Time Limited Visual Search associated with raking through leaf litter. We recorded 20 species from 10 families and used non‐metric multidimensional scaling to reduce the dimensionality of quantitative and qualitative compositions of species. Multiple linear regression models indicated that the environmental gradients distance to nearest stream, extent of canopy openness, vegetation density and slope did not significantly influence assemblage species distribution, with an indication of effect of litter depth. By means of piecewise linear regression, the use of riparian zone was estimated at ~190 m from quantitative species composition and ~211 m from qualitative species composition. Five species occurred only in the riparian zone. Our results suggest that conservation of the entire riparian lizard assemblage in Amazonian rainforest is likely to require protection of at least a 211 m buffer on either side of streams.     

A Matter of Scale: Historical and Environmental Factors Structure Anuran Assemblages from the Upper Madeira River,Amazonia

Biogeographical history and current ecological interactions have usually been addressed separately to explain the spatial distribution of patterns of biodiversity. In this study, we evaluated the integrated effects of biogeographical and environmental factors in structuring the diurnal amphibian anuran assemblages of the upper Madeira River, southwestern Amazonia. We used a sampling design involving 98 standardized units, distributed across seven locations covering both banks of the river's course in the state of Rondônia, Brazil. We conducted searches for frogs in three campaigns between February 2010 and February 2011, aiming to: (1) evaluate the effect of the Madeira River as a biogeographic barrier at the species‐assemblage level, and (2) test the influence of seven environmental variables (vegetation structure, vegetation cover, soil nutrients, soil structure, slope, elevation, and distance from the river bank) on the spatial structure of the frog assemblages, separately on each riverbank. Thirteen species of diurnal frogs were recorded, six of which were restricted to one of the river margins. Multivariate analysis of variance indicated a significant effect of the river as a barrier. Multiple regression analyses suggested that the environmental variables structuring frog assemblages differ on either side of the river. We found that both historical elements (on a regional scale) and environmental factors (at a local scale) shaped the occurrence and distribution of frog species in the study area.     

Beyond the river: underlying determinants of population acoustic signal variability in Amazonian direct-developing Allobates (Anura: Dendrobatoidea)

The multidimensional nature of animal signals makes acoustic traits potentially subject to different determinants. The Amazonian frogs Allobates nidicola and Allobates masniger have an allopatric distribution, occurring along the left and right sides of the Madeira River, respectively. These are two sister, phenotypically similar species whose eggs are deposited and develop entirely in a terrestrial nest. In this study, we analyzed 2,000 advertisement calls recorded from ten localities across Central Amazonia, in order to understand the role of determinants of acoustic signal variability at the population and species levels. We assessed, through nested analyses of variance, the differentiation of six characters of this sexual signal among populations and between interfluves. Moreover, we measured the degree of variability and the extent of temperature- and body size-induced plasticity in call traits. We also tested for isolation-by-distance effects in phenotypic differentiation through Mantel tests. Coefficients of variation were higher among than within populations for all call measurements. Spectral call properties were more distinctive than temporal traits among populations and species. Advertisement call traits showed strong temperature-induced plasticity (e.g., 45?% of the variation in note duration). In contrast, the effects of body size were restricted to frequency-related characters. The river barrier effect was significant among all the acoustic variables analyzed even after controlling for male body size. Geography (sampling locality) and body size also jointly affected call variability. No correlation between geographical and acoustic distances among populations was observed, suggesting that local stabilizing selective pressures have an important role in the evolution of call differentiation.     

Microarray analysis of HIV resistant female sex workers reveal a gene expression signature pattern reminiscent of a lowered immune activation state

To identify novel biomarkers for HIV-1 resistance, including pathways that may be critical in anti-HIV-1 vaccine design, we carried out a gene expression analysis on blood samples obtained from HIV-1 highly exposed seronegatives (HESN) from a commercial sex worker cohort in Nairobi and compared their profiles to HIV-1 negative controls. Whole blood samples were collected from 43 HIV-1 resistant sex workers and a similar number of controls. Total RNA was extracted and hybridized to the Affymetrix HUG 133 Plus 2.0 micro arrays (Affymetrix, Santa Clara CA). Output data was analysed through ArrayAssist software (Agilent, San Jose CA). More than 2,274 probe sets were differentially expressed in the HESN as compared to the control group (fold change ≥1.3; p value ≤0.0001, FDR <0.05). Unsupervised hierarchical clustering of the differentially expressed genes readily distinguished HESNs from controls. Pathway analysis through the KEGG signaling database revealed a majority of the impacted pathways (13 of 15, 87%) had genes that were significantly down regulated. The most down expressed pathways were glycolysis/gluconeogenesis, pentose phosphate, phosphatidyl inositol, natural killer cell cytotoxicity and T-cell receptor signaling. Ribosomal protein synthesis and tight junction genes were up regulated. We infer that the hallmark of HIV-1 resistance is down regulation of genes in key signaling pathways that HIV-1 depends on for infection.     

Territory size as a main driver of male-mating success in an Amazonian nurse frog (<Emphasis Type="Italic">Allobates paleovarzensis</Emphasis>, Dendrobatoidea)

In polygamous mating systems, it is most often males that compete for the opposite sex, using strategies to provide access to as many females as possible. Females, on the other hand, constitute the sex that exerts the choice and so require a means of accessing the quality of a potential partner in comparison to its competitors. A common challenge in sexual selection studies is to identify the most relevant trait for mating success, since many are correlated with each other. In addition, little is known about how the female accesses the aspects related to male quality. In this context, we tested the role of different male characteristics on mating success in a natural environment using the Amazonian frog Allobates paleovarzensis as a model. A multiple linear regression model showed a positive relationship between territory size and number of male matings, while calling persistence was slightly related to the mating success. We did not detect a relation of the number of matings with the distance to the nearest body of water nor with male body size. Additionally, we observed that territory size was not related to calling persistence, but had a positive relation with the duration of the couple’s courting process. Thus, we conclude that: (1) territory size is the main determinant of male-mating success, and this is not correlated with the other attributes tested; and (2) females access the size of the males’ territory through the courting process that precedes oviposition.     

A novel in-vitro system for the simultaneous exposure of bladder smooth muscle cells to mechanical strain and sustained hydrostatic pressure

The novel hydrostrain system was designed in an effort to establish and maintain conditions that simulate the in-vivo mechanical environment of the bladder. In this laboratory system, ovine bladder smooth muscle cells on flexible, 10-cm-dia silastic membranes were exposed simultaneously to hydrostatic pressure (40 cm H2O, a pressure level currently associated with bladder pathologies) and mechanical strains (up to 25 percent) under standard cell culture conditions for 7 h. Under these conditions, Heparin Binding-Epidermal Growth Factor and Collagen Type III mRNA expression were significantly increased (p<0.01 and 0.1, respectively); however, no changes were observed in Collagen Type I mRNA expression. Decreases in the Collagen Type I:Type III ratio following simultaneous exposure of bladder smooth muscle cells to pathological levels of hydrostatic pressure and mechanical strain in vitro are in agreement with clinically observed increases in Collagen Type III with concomitant decreased human bladder compliance. The results of the present study, therefore, provide cellular/molecular level information relevant to bladder pathology that could have significant implications in the field of clinical urology.     

p38 MAPK mediates renal tubular cell TNF-{alpha} production and TNF-{alpha}-dependent apoptosis during simulated ischemia

Ischemia causes renal tubular cellloss through apoptosis; however, the mechanisms of this processremain unclear. Using the renal tubular epithelial cell lineLLC-PK₁, we developed a model of simulated ischemia(SI) to investigate the role of p38 MAPK (mitogen-activated proteinkinase) in renal cell tumor necrosis factor- (TNF-) mRNAproduction, protein bioactivity, and apoptosis. Resultsdemonstrate that 60 min of SI induced maximal TNF- mRNA productionand bioactivity. Furthermore, 60 min of ischemia induced renaltubular cell apoptosis at all substrate replacement time pointsexamined, with peak apoptotic cell death occurring after either 24 or 48 h. p38 MAPK inhibition abolished TNF- mRNA production andTNF- bioactivity, and both p38 MAPK inhibition and TNF- neutralization (anti-porcine TNF- antibody) preventedapoptosis after 60 min of SI. These results constitute theinitial demonstration that 1) renal tubular cells produceTNF- mRNA and biologically active TNF- and undergoapoptosis in response to SI, and 2) p38 MAPKmediates renal tubular cell TNF- production and TNF--dependent apoptosis after SI.

     

Sampling design may obscure species–area relationships in landscape-scale field studies

We investigated 1) the role of area per se in explaining anuran species richness on reservoir forest islands, after controlling for several confounding factors. We also assessed 2) how sampling design affects the inferential power of island species–area relationships (ISARs) aiming to 3) provide guidelines to yield reliable estimates of area-induced species losses in patchy systems. We surveyed anurans with autonomous recording units at 151 plots located on 74 islands and four continuous forest sites at the Balbina Hydroelectric Reservoir landscape, central Brazilian Amazonia. We applied semi-log ISAR models to assess the effect of sampling design on the fit and slope of species–area curves. To do so, we subsampled our surveyed islands following both a 1) stratified and 2) non-stratified random selection of 5, 10, 15, 20 and 25 islands covering 1) the full range in island size (0.45–1699 ha) and 2) only islands smaller than 100 ha, respectively. We also compiled 25 datasets from the literature to assess the generality of our findings. Island size explained ca half of the variation in species richness. The fit and slope of species–area curves were affected mainly by the range in island size considered, and to a very small extent by the number of islands surveyed. In our literature review, all datasets covering a range of patch sizes larger than 300 ha yielded a positive ISAR, whereas the number of patches alone did not affect the detection of ISARs. We conclude that 1) area per se plays a major role in explaining anuran species richness on forest islands within an Amazonian anthropogenic archipelago; 2) the inferential power of island species–area relationships is severely degraded by sub-optimal sampling designs; 3) at least 10 habitat patches spanning three orders of magnitude in size should be surveyed to yield reliable species–area estimates in patchy systems.     

A Comparative Analysis of Gene Expression Patterns and Cell Phenotypes between Cervical and Peripheral Blood Mononuclear Cells

Studies of the immunological environment in the female genital tract (FGT) are critical for the development of vaccines or microbicides to halt the spread of sexually transmitted infections. Challenges arise due to the difficulties of sampling from this site, and the majority of studies have been conducted utilising peripheral blood mononuclear cells. Identifying functional differences between immune cells of the FGT and peripheral blood would aid in our understanding of mucosal immunology. We compared the gene expression profile of mononuclear cells at these two sites. Messenger RNA expression analysis was performed using gene expression arrays on matched cervical mononuclear cells and peripheral blood mononuclear cells. Further cellular phenotyping was done by 10 colour flow cytometry. Of the 22,185 genes expressed by these samples, 5345 genes were significantly differentially expressed between the cell populations. Most differences can be explained by significantly lower levels of T and B cells and higher levels of macrophages and dendritic cells in the FGT compared with peripheral blood. Several immunologically relevant pathways such as apoptosis and innate immune signalling, and a variety of cytokines and cytokine receptors were differentially expressed. This study highlights the importance of the unique immunological environment of the FGT and identifies important differences between systemic and mucosal immune compartments.     

The Early Stages of Speciation in Amazonian Forest Frogs: Phenotypic Conservatism Despite Strong Genetic Structure

Phylogeographic perspectives incorporating multiple classes of characters, especially those relating to sexual signals, are promising for the elucidation of recent evolutionary mechanisms driving speciation. Here, forest frogs were used as a model system to access distinct stages in the process of evolutionary differentiation. We studied 280 individuals assigned to three species: Allobates paleovarzensis, A. nidicola and A. masniger. Samples were collected at 20 localities arranged in two study systems, along the middle Amazon and the lower Madeira Rivers, in Central Amazonia. Mantel tests, analyses of molecular variance, and the spatial distribution of haplogroups indicated that the distribution of genetic variability, as inferred from a mitochondrial DNA marker, was determined by a combination of isolation-by-distance effects and the transposition of large Amazonian rivers. These two factors had contrasting relative influences in each of the study systems, which also differed regarding the estimated time of the major cladogenetic events. Pronounced population genetic structure was observed. However, multivariate discriminant function analyses revealed that the phenotypic (morphological and acoustic) divergence was loosely related with genetic differentiation and did not successfully predict assignment of individuals to genetic groups. The observed distribution of genetic variability showed the important role of genetic drift in the diversification of the mitochondrial marker studied. The phenotypic conservatism among populations was surprising in view of the high genetic structuring observed, and indicates a prevailing role of stabilizing selective forces in the process of sexual signal and morphological differentiation.