Exploring the Distribution of Genetic Markers of Pharmacogenomics Relevance in Brazilian and Mexican Populations

Studies of pharmacogenomics-related traits are increasingly being performed to identify loci that affect either drug response or susceptibility to adverse drug reactions. However, the effect of the polymorphisms can differ in magnitude or be absent depending on the population being assessed. We used the Affymetrix Drug Metabolizing Enzymes and Transporters (DMET) Plus array to characterize the distribution of polymorphisms of pharmacogenetics and pharmacogenomics (PGx) relevance in two samples from the most populous Latin American countries, Brazil and Mexico. The sample from Brazil included 268 individuals from the southeastern state of Rio de Janeiro, and was stratified into census categories. The sample from Mexico comprised 45 Native American Zapotecas and 224 self-identified Mestizo individuals from 5 states located in geographically distant regions in Mexico. We evaluated the admixture proportions in the Brazilian and Mexican samples using a panel of Ancestry Informative Markers extracted from the DMET array, which was validated with genome-wide data. A substantial variation in ancestral proportions across census categories in Brazil, and geographic regions in Mexico was identified. We evaluated the extent of genetic differentiation (measured as F_ST values) of the genetic markers of the DMET Plus array between the relevant parental populations. Although the average levels of genetic differentiation are low, there is a long tail of markers showing large frequency differences, including markers located in genes belonging to the Cytochrome P450, Solute Carrier (SLC) and UDP-glucuronyltransferase (UGT) families as well as other genes of PGx relevance such as ABCC8, ADH1A, CHST3, PON1, PPARD, PPARG, and VKORC1. We show how differences in admixture history may have an important impact in the distribution of allele and genotype frequencies at the population level.     

In vivo nanoscale analysis of the dynamic synergistic interaction of Bacillus thuringiensis Cry11Aa and Cyt1Aa toxins in Aedes aegypti

The insecticidal Cry11Aa and Cyt1Aa proteins are produced by Bacillus thuringiensis as crystal inclusions. They work synergistically inducing high toxicity against mosquito larvae. It was proposed that these crystal inclusions are rapidly solubilized and activated in the gut lumen, followed by pore formation in midgut cells killing the larvae. In addition, Cyt1Aa functions as a Cry11Aa binding receptor, inducing Cry11Aa oligomerization and membrane insertion. Here, we used fluorescent labeled crystals, protoxins or activated toxins for in vivo localization at nano-scale resolution. We show that after larvae were fed solubilized proteins, these proteins were not accumulated inside the gut and larvae were not killed. In contrast, if larvae were fed soluble non-toxic mutant proteins, these proteins were found inside the gut bound to gut-microvilli. Only feeding with crystal inclusions resulted in high larval mortality, suggesting that they have a role for an optimal intoxication process. At the macroscopic level, Cry11Aa completely degraded the gastric caeca structure and, in the presence of Cyt1Aa, this effect was observed at lower toxin-concentrations and at shorter periods. The labeled Cry11Aa crystal protein, after midgut processing, binds to the gastric caeca and posterior midgut regions, and also to anterior and medium regions where it is internalized in ordered “net like” structures, leading finally to cell break down. During synergism both Cry11Aa and Cyt1Aa toxins showed a dynamic layered array at the surface of apical microvilli, where Cry11Aa is localized in the lower layer closer to the cell cytoplasm, and Cyt1Aa is layered over Cry11Aa. This array depends on the pore formation activity of Cry11Aa, since the non-toxic mutant Cry11Aa-E97A, which is unable to oligomerize, inverted this array. Internalization of Cry11Aa was also observed during synergism. These data indicate that the mechanism of action of Cry11Aa is more complex than previously anticipated, and may involve additional steps besides pore-formation activity.     

Eukaryotic ectosymbionts of Acari

The Acari is the most numerous and diverse group of the subphylum Chelicerata. With approximately 55 000 described species (and estimates of up to 1 million extant species), their adaptations for parasitism, phytophagy, mycophagy, saprophagy and predation rival other arthropods and challenge us with a wide variety of biological interactions. While a few studies have unravelled the nature of some endosymbiotic associations between mites or ticks and prokaryotes, almost nothing has been done yet regarding acarine eukaryotic ectosymbionts. Microbial ectosymbionts can benefit their hosts by providing nutrients, by aiding digestion, by enhancing communication, by assisting in mating and/or fertilization, by protecting their host against pathogenic microorganisms, against predation and so on. In this sketch, we introduce a number of described cases of fungal and protist ectosymbionts and discuss the role they might play in the life of their acarine hosts.     

Isolation of Cry1Ab protein mutants of Bacillus thuringiensis by a highly efficient PCR site-directed mutagenesis system

Abstract A site-directed mutagenesis method was designed and used to create Cry1Ab mutant proteins in two of the five highly conserved blocks present in the Cry protein family. Region 1 comprises the central α-helix 5 of domain I and has been implicated in the pore formation activity of the toxin. Substitution of arginine by serine at position 173 (R173S) affects neither structural integrity nor toxicity. Region 2 comprises the major part of the domain I/domain II interface, characterized by the presence of numerous hydrogen bonds and electrostatic interactions. Mutations in the salt bridge formed by aspartic acid 242 and arginine 265 (D242N, D242C, R265C, and D242C/R265C) resulted in structurally unstable mutant proteins as is shown by their increased protease sensitivity and lack of biological activity.     

Testing the abundant‐centre hypothesis using intertidal porcelain crabs along the Chilean coast: linking abundance and life‐history variation

Aim The abundant‐centre hypothesis (ACH) is based on the assumption that physiological constraints limit populations at the edges of their distributional range, yet the geographical variation of physiological performance or life‐history traits has rarely been examined. Here we examine the applicability of the ACH in a marine system by testing whether physiological predictions are reflected in large‐scale variations of life‐history traits. Location The Chilean coast (18°–42° S), encompassing more than 2500 km along the Pacific coast of South America. Methods Five porcelain crab species (Petrolisthes granulosus, Petrolisthes laevigatus, Petrolisthes tuberculatus, Petrolisthes violaceus and Allopetrolisthes angulosus) were sampled on intertidal boulder beaches at 13 sampling sites. For each species and site we evaluated: (1) relative abundance (density), (2) maximum size, (3) size at maturity, (4) sex ratio, (5) proportion of ovigerous females, and (6) presence of recruits. The shape of the spatial distribution of each trait was evaluated statistically against the prediction of four hypothetical models (normal, ramped‐south, ramped‐north and abundant‐edge). Results The relative abundance and life‐history traits showed different spatial patterns among species. Relative abundance (across sites) was fitted by a normal model in only two species. No model fitted the spatial variation in body size and size at first maturity, which showed a slight but monotonic poleward increase in all species. Sex ratio showed a prominent hump‐shaped pattern, with females prevailing in the centre of the ranges and males dominating towards the range boundaries; this pattern was statistically significant in three of the five studied species. The proportion of ovigerous females showed no clear latitudinal trends, and mature individuals were observed across most of the geographical range of the species. However, recruits tended to be absent towards the southern (poleward) boundaries of the distribution. Main conclusions The ACH does not apply to all species equally. The link between abundance and life‐history traits is complex and variable among the porcelain crab species studied. Overall, the observed patterns were consistent with the idea that equatorward boundaries might be controlled by physiological restrictions mainly affecting adult survival, whereas poleward boundaries might be shaped by limitations in reproductive output and larval survival. Our results underline the importance of incorporating ecological, physiological and life‐history studies in future tests of the ACH.     

Effect of chronic lithium on mechanical sensitivity and trabecular bone loss induced by type-1 diabetes mellitus in mice

BioMetals - Type-1 diabetes mellitus (T1DM) is a chronic condition characterized by long-term hyperglycemia that results in several complications such as painful peripheral neuropathy, bone...     

Do brassinosteroids alleviate water stress in seedlings of Handroanthus serratifolius?

Limited water availability in reforestation areas can compromise plant growth and development, especially for plants at early stages. In this context, the exogenous application of growth regulators, such as brassinosteroids (BRs), can be used to mitigate the negative effects of water restriction. The aim of this study was to evaluate the effect of the application of BRs (90% brassinolide + a brassinosteroid) on initial growth, gas exchange, leaf water potential and leaf anatomy in yellow Ipê (Handroanthus serratifolius), under water restriction conditions. Throughout the experimental period, plants were maintained in two different water regimes: WR1—plants maintained at 100% of the pot capacity; WR2—plants rehydrated with 50% of the evapotranspiration from the previous day. BRs were applied on three occasions during the experimental period, in five concentrations: C1, 0 g/L (application of distilled water); C2, 0.25 g/L; C3, 0.5 g/L; C4, 0.75 g/L; and C5, 1.00 g/L. The dry mass, root:shoot ratio, water status and leaf anatomy traits were evaluated at the end of the experimental period and were analysed in two-level factorial scheme (2WR × 5BRs concentrations). The plant height, number of leaves, gas exchange and leaf chlorophyll content were evaluated four times during the experimental period and were analysed in a split-split-plot design (two WR × five BRs concentrations × four evaluation times). The differences between the means were evaluated by analysis of variance (ANOVA). Pairs of means were separated using the standard error of the difference between the means (SED) and the Fisher's protected least significant difference test (LSD) at p < .05. In addition, all variables were subjected to regression analysis, being the variables evaluated over time, analysed through a two-stage modelling approach. The results obtained indicate that the water deficit led to reductions in growth and gas exchange parameters, regardless of the exogenous application of the regulator; therefore, higher concentrations (C4 and C5) were the most harmful for the maintenance of metabolic and photosynthetic activity. The exogenous application of BRs in H. serratifolius plants attenuated the effects of water limitation on the leaf water potential, but was not able to mitigate for the negative effects in growth, gas exchange and leaf anatomy.