Pharmacophore modeling,docking and molecular dynamics to identify <Emphasis Type="Italic">Leishmania major</Emphasis> farnesyl pyrophosphate synthase inhibitors

Leishmaniasis is caused by protozoa of the genus Leishmania spp. and is considered the second most important protozoa in the world due to the number of cases and mortality. Despite its importance in terms of public health, the treatment of patients is limited and has mostly low levels of efficacy and safety. Farnesyl pyrophosphate synthase (FPPS) acts in the early stages of isoprenoid synthesis, and is important for maintaining the integrity of the lipid bilayer of the parasite that causes the disease. The aim of this work was to identify one potential inhibitor of the FPPS of Leishmania major through virtual screening by pharmacophore modeling and docking. A total of 85,000 compounds from a natural products database (ZINC¹⁵) was submitted for virtual hierarchical screening, and the top ranked molecule in both methods was analyzed by intermolecular interaction profile and 20 ns molecular dynamics simulations. These results showed a promising compound from natural products that mimic the major interactions present in the substrate/inhibitor.     

CO2 emission from mineral soils following land‐cover change in Brazil

This paper presents a first approximation of net CO₂ fluxes from mineral soil due to land use, land‐use changes and forestry (LULUCF) activities in Brazil for the periods 1970–90 and 1975–95. The methodology employed is an adaptation of the approach proposed by the IPCC in ‘Revised 1996 guidelines for national greenhouse gas inventories’, which is based on the variations in soil C stocks as a function of changes in land‐use activities. The calculation was done separately for each Brazilian state and subsequently summarized for all of Brazil. The annual fluxes for Brazil indicate a net emission of CO₂ to the atmosphere, which decreased from 93.3 Tg CO₂ for the period 1970–90 to 46.4 Tg CO₂ for the period 1975–95. This corresponded to yearly net emission rates of 11.0 g CO₂ m^?2 year^?1 for the 1970–90 period and 5.5 g CO₂ m^?2 year^?1 for the 1975–95 period. Within each administrative region, considerable differences in the yearly emission rates between the states could be observed. Several sources of uncertainties could be identified. The most important uncertainties were linked to the impact factor values, which represent changes in native C stock associated with conversion of the native vegetation to agricultural use.     

Host specificity of Metriona elatior, a potential biological control agent of tropical soda apple, Solanum viarum, in the USA

The leaf beetle Metriona elatior from Brazil-Argentina was screened in the Florida (USA)State quarantine facility as a potential biological control agent of tropical soda apple, Solanum viarum, a recently arrived weed species. Multiple-choice host-specificity tests were conducted in small cages (60 cm × 60 cm × 60 cm) using 95 plant species in 29 families. Adults fed heavily on the main target weed (S. viarum), and on turkey berry,Solanum torvum (noxious weed of Asiatic origin); fed moderately on red soda apple, Solanum capsicoides (weed of South American origin), and eggplant, Solanum melongena (economic crop); and fed lightly on aquatic soda apple, Solanum tampicense (weed of Mexican-Caribbean-Central American origin), and onsilverleaf nightshade, Solanum elaeagnifolium(native weed widely distributed). M.elatior adults laid 84 to 97% of their egg masses onS. viarum, and 3 to 16% on S. melongena. Non-choice host-specificity tests were also conducted in quarantine in which M. elatior adults and neonate larvae were exposed to 17 and 19 plant species, respectively. Tests with the neonates indicate that this insect was able to complete its development on S. viarum, S. torvum, S. melongena, and S. capsicoides. Although some adult feeding and oviposition occurred on S.melongena in quarantine on potted plants in small cages, no feeding or oviposition by M. elatiorwas observed in field experiments conducted in Brazil. Surveys in unsprayed S. melongena fields in Argentina and Brazil indicated that M. elatioris not a pest of S. melongena in South America. The evidence obtained from the South-American field surveys, Brazil open-field experiments, and Florida quarantine host specificity tests indicate that M. elatior causes significant feeding damage toS. viarum, and does not represent a threat to S. melongena crops in the USA. Therefore an application for permission to releaseM. elatior against S. viarum in the USA was submitted in October 1998. This revised version was published online in July 2006 with corrections to the Cover Date.     

Synthesis and biological evaluation of 1,2-disubstituted carbonucleosides of 6-substituted purine and 8-azapurine.

A series of new one two substituted carbonucleoside analogues (OTC), with the purine and 8-azapurine base linked through a methylene group at the cyclopentane ring, were synthesized and evaluated for their activity against a number of viruses and tumor cells in vitro.     

Functional differences in echolocation call design in an adaptive radiation of bats

All organisms have specialized systems to sense their environment. Most bat species use echolocation for navigation and foraging, but which and how ecological factors shaped echolocation call diversity remains unclear for the most diverse clades, including the adaptive radiation of neotropical leaf‐nosed bats (Phyllostomidae). This is because phyllostomids emit low‐intensity echolocation calls and many inhabit dense forests, leading to low representation in acoustic surveys. We present a field‐collected, echolocation call dataset spanning 35 species and all phyllostomid dietary guilds. We analyze these data under a phylogenetic framework to test the hypothesis that echolocation call design and parameters are specialized for the acoustic demands of different diets, and investigate the contributions of phylogeny and body size to echolocation call diversity. We further link call parameters to dietary ecology by contrasting minimum detectable prey size estimates (MDPSE) across species. We find phylogeny and body size explain a substantial proportion of echolocation call parameter diversity, but most species can be correctly assigned to taxonomic (61%) or functional (77%) dietary guilds based on call parameters. This suggests a degree of acoustic ecological specialization, albeit with interspecific similarities in call structure. Theoretical MDPSE are greatest for omnivores and smallest for insectivores. Omnivores significantly differ from other dietary guilds in MDPSE when phylogeny is not considered, but there are no differences among taxonomic dietary guilds within a phylogenetic context. Similarly, predators of non‐mobile/non‐evasive prey and predators of mobile/evasive prey differ in estimated MDPSE when phylogeny is not considered. Phyllostomid echolocation call structure may be primarily specialized for overcoming acoustic challenges of foraging in dense habitats, and then secondarily specialized for the detection of food items according to functional dietary guilds. Our results give insight into the possible ecological mechanisms shaping the diversity of sensory systems, and their reciprocal influence on resource use.     

Docosahexaenoic acid in diluent for goat semen cryopreservation

The effects of docosahexaenoic acid (DHA) in the diluent for cryopreservation of goat semen on seminal quality and the optimal levels to be used were evaluated. After collection, semen was pooled and physically evaluated, then divided into four aliquots with different DHA levels in the diluent: 0, 10, 20, and 30 ng mL^-1. The semen was cryopreserved in a TK 3000® freezing machine and then thawed for assessment at 37 °C. Sperm motility and vigor, membrane integrity, acrosomal integrity, mitochondrial activity, and sperm chromatin compaction were evaluated after thawing. A completely randomized design was used. For normally distributed variables, ANOVA and regression analysis were used to test for differences between treatments, and for non-parametric data, the Kruskal Wallis test was used at the 5% significance level. There were no differences among groups in terms of membrane integrity, acrosomal integrity, or chromatin compaction. There was a decrease in class I mitochondrial activity with increasing DHA level (P<0.05), but no differences in classes II, III, and IV (P>0.05). The inclusion of 10 to 30 ng mL^-1 of DHA in the diluent did not result in improvements in seminal quality parameters after thawing, with some impairment observed in the mitochondrial activity of the sperm cells.     

Dynamic L-type CaV1.2 channel trafficking facilitates CaV1.2 clustering and cooperative gating

L-type Ca_V1.2 channels are key regulators of gene expression, cell excitability and muscle contraction. Ca_V1.2 channels organize in clusters throughout the plasma membrane. This channel organization has been suggested to contribute to the concerted activation of adjacent Ca_V1.2 channels (e.g. cooperative gating). Here, we tested the hypothesis that dynamic intracellular and perimembrane trafficking of Ca_V1.2 channels is critical for formation and dissolution of functional channel clusters mediating cooperative gating. We found that Ca_V1.2 moves in vesicular structures of circular and tubular shape with diverse intracellular and submembrane trafficking patterns. Both microtubules and actin filaments are required for dynamic movement of Ca_V1.2 vesicles. These vesicles undergo constitutive homotypic fusion and fission events that sustain Ca_V1.2 clustering, channel activity and cooperative gating. Our study suggests that Ca_V1.2 clusters and activity can be modulated by diverse and unique intracellular and perimembrane vesicular dynamics to fine-tune Ca²⁺ signals.     

Establishment and characterization of human bladder cancer cell lines BexBra1, BexBra2, and BexBra4

Bladder cancer (BC) is the fourth most common cancer in the USA. In Brazil, BC represents 3% of the total existing carcinomas in the population and represents the second highest incidence among urological tumors. The majority of bladder cancer cell lines available were derived from Caucasians and established in the seventies or eighties. Thus, neoplasia development in these cells likely occurred in environment conditions vastly different than today. In the present study, we report the establishment and characterization of three Brazilian bladder cancer cell lines (BexBra1, BexBra2, and BexBra4). These cell lines may be helpful for dissecting the genetic and epigenetic aspects that trigger the progression of BC. Moreover, the development of a Brazilian representative of the disease will allow us to investigate the potential inter-racial differences of malignancy-associated phenotypes in bladder cancer.     

Nonlinearity and fractality in the variability of cardiac period in the lizard, Gallotia galloti: effects of autonomic blockade

Both nonlinear and fractal properties of beat-to-beat R-R interval variability signal (RRV) of freely moving lizards (Gallotia galloti) were studied in baseline and under autonomic nervous system blockade. Nonlinear techniques allowed us to study the complexity, chaotic behavior, nonlinearity, stationarity, and regularity over time of RRV. Scaling behavior of RRV was studied by means of fractal techniques. The autonomic nervous system blockers used were atropine, propranolol, prazosin, and yohimbine. The nature of RRV was linear in baseline and under beta-, alpha(1)- and alpha(2)-adrenoceptor blockades. Atropine changed the linear nature of RRV to nonlinear and increased its stationarity, regularity and fractality. Propranolol increased the complexity and chaotic behavior, and decreased the stationarity, regularity, and fractality of RRV. Both prazosin and yohimbine did not change any of the nonlinear and fractal properties of RRV. It is suggested that 1) the use of both nonlinear and fractal analysis is an appropriate approach for studying cardiac period variability in reptiles; 2) the cholinergic activity, which seems to make the alpha(1)-, alpha(2)- and beta-adrenergic activity interaction unnecessary, determines the linear behavior in basal RRV; 3) fractality, as well as both RRV regularity and stationarity over time, may result from the balance between cholinergic and beta-adrenergic activities opposing actions; 4) beta-adrenergic activity may buffer both the complexity and chaotic behavior of RRV, and 5) neither the alpha(1)- nor the alpha(2)-adrenergic activity seem to be involved in the mediation of either nonlinear or fractal components of RRV.     

Analysis of the Paracoccidioides brasiliensis triosephosphate isomerase suggests the potential for adhesin function

Paracoccidioides brasiliensis is an important fungal pathogen. The disease it causes, paracoccidioidomycosis (PCM), ranges from localized pulmonary infection to systemic processes that endanger the life of the patient. Paracoccidioides brasiliensis adhesion to host tissues contributes to its virulence, but we know relatively little about molecules and the molecular mechanisms governing fungal adhesion to mammalian cells. Triosephosphate isomerase (TPI: EC 5.3.1.1) of P. brasiliensis (PbTPI) is a fungal antigen characterized by microsequencing of peptides. The protein, which is predominantly expressed in the yeast parasitic phase, localizes at the cell wall and in the cytoplasmic compartment. TPI and the respective polyclonal antibody produced against this protein inhibited the interaction of P. brasiliensis to in vitro cultured epithelial cells. TPI binds preferentially to laminin, as determined by peptide inhibition assays. Collectively, these results suggest that TPI is required for interactions between P. brasiliensis and extracellular matrix molecules such as laminin and that this interaction may play an important role in the fungal adherence and invasion of host cells.