Functional characterization of a synthetic hydrophilic antifungal peptide derived from the marine snail Cenchritis muricatus

Antimicrobial peptides have been found in mollusks and other sea animals. In this report, a crude extract of the marine snail Cenchritis muricatus was evaluated against human pathogens responsible for multiple deleterious effects and diseases. A peptide of 1485.26 Da was purified by reversed-phase HPLC and functionally characterized. This trypsinized peptide was sequenced by MS/MS technology, and a sequence (SRSELIVHQR), named Cm-p1 was recovered, chemically synthesized and functionally characterized. This peptide demonstrated the capacity to prevent the development of yeasts and filamentous fungi. Otherwise, Cm-p1 displayed no toxic effects against mammalian cells. Molecular modeling analyses showed that this peptide possible forms a single hydrophilic α-helix and the probable cationic residue involved in antifungal activity action is proposed. The data reported here demonstrate the importance of sea animals peptide discovery for biotechnological tools development that could be useful in solving human health and agribusiness problems.     

Genetic characterization and construction of an auxotrophic strain of Saccharomyces cerevisiae JP1, a Brazilian industrial yeast strain for bioethanol production

Used for millennia to produce beverages and food, Saccharomyces cerevisiae also became a workhorse in the production of biofuels, most notably bioethanol. Yeast strains have acquired distinct characteristics that are the result of evolutionary adaptation to the stresses of industrial ethanol production. JP1 is a dominant industrial S. cerevisiae strain isolated from a sugarcane mill and is becoming increasingly popular for bioethanol production in Brazil. In this work, we carried out the genetic characterization of this strain and developed a set of tools to permit its genetic manipulation. Using flow cytometry, mating type, and sporulation analysis, we verified that JP1 is diploid and homothallic. Vectors with dominant selective markers for G418, hygromycin B, zeocin, and ρ-fluoro-dl-phenylalanine were used to successfully transform JP1 cells. Also, an auxotrophic ura3 mutant strain of JP1 was created by gene disruption using integration cassettes with dominant markers flanked by loxP sites. Marker excision was accomplished by the Cre/loxP system. The resulting auxotrophic strain was successfully transformed with an episomal vector that allowed green fluorescent protein expression.     

Molecular detection of XO - Turner syndrome

Turner syndrome is caused by haploinsufficiency of the short arm of X-chromosome, and is usually diagnosed by karyotyping. This procedure is time-consuming, expensive and unfeasible for population screening. We propose molecular detection of 45XO Turner patients based on the ability of HpaII, a methylation sensitive endonuclease, to induce the cleavage of non-methylated DNA in the active X-allele. Genomic DNA was obtained from 22 patients with Turner syndrome confirmed by karyotype (45XO, N = 18; 45XO/46XX, N = 4). After digestion, DNA was amplified with primers directed to exon 1 of the androgen receptor (AR) gene and to the GAPDH control gene. Normal control females or mosaic patients, with a second methylated X-chromosome, escaped from HpaII digestion and produced a band corresponding to AR gene amplification. 45XO patients have just one active non-methylated X-chromosome, completely digested by HpaII, thus preventing the amplification of the AR gene. Three of the 45XO cases gave amplified bands, suggesting low-frequency mosaicisms that are not detected by karyotyping. Compared to classical karyotype studies for the detection of 45XO Turner patients, this new molecular method is simpler, faster and less expensive.     

Determinants of reef fish assemblages in tropical Oceanic islands

Diversity patterns are determined by biogeographic, energetic, and anthropogenic factors, yet few studies have combined them into a large‐scale framework in order to decouple and compare their relative effects on fish faunas. Using an empirical dataset derived from 1527 underwater visual censuses (UVC) at 18 oceanic islands (five different marine provinces), we determined the relative influence of such factors on reef fish species richness, functional dispersion, density and biomass estimated from each UVC unit. Species richness presented low variation but was high at large island sites. High functional dispersion, density, and biomass were found at islands with large local species pool and distance from nearest reef. Primary productivity positively affected fish richness, density and biomass confirming that more productive areas support larger populations, and higher biomass and richness on oceanic islands. Islands densely populated by humans had lower fish species richness and biomass reflecting anthropogenic effects. Species richness, functional dispersion, and biomass were positively related to distance from the mainland. Overall, species richness and fish density were mainly influenced by biogeographical and energetic factors, whereas functional dispersion and biomass were strongly influenced by anthropogenic factors. Our results extend previous hypotheses for different assemblage metrics estimated from empirical data and confirm the negative impact of humans on fish assemblages, highlighting the need for conservation of oceanic islands.     

Biogeographic responses and niche occupancy of microbial communities following long-term land-use change

Understanding the effects of forest-to-agriculture conversion on microbial diversity has been a major goal in soil ecological studies. However, linking community assembly to the ruling ecological processes at local and regional scales remains challenging. Here, we evaluated bacterial community assembly patterns and the ecological processes governing niche specialization in a gradient of geography, seasonality, and land-use change, totaling 324 soil samples, 43 habitat characteristics (abiotic factors), and 16 metabolic and co-occurrence patterns (biotic factors), in the Brazilian Atlantic Rainforest, a subtropical biome recognized as one the world’s largest and most threatened hotspots of biodiversity. Pairwise beta diversities were lower in pastures than in forest and no-till soils. Pasture communities showed a predominantly neutral model, regarding stochastic processes, with moderate dispersion, leading to biotic homogenization. Most no-till and forest microbial communities followed a niche-based model, with low rates of dispersal and weak homogenizing selection, indicating niche specialization or variable selection. Historical and evolutionary contingencies, as represented by soil type, season, and dispersal limitation were the main drivers of microbial assembly and processes at the local scale, markedly correlated with the occurrence of endemic microbes. Our results indicate that the patterns of assembly and their governing processes are dependent on the niche occupancy of the taxa evaluated (generalists or specialists). They are also more correlated with historical and evolutionary contingencies and the interactions among taxa (i.e., co-occurrence patterns) than the land-use change itself.

     

The role of effective mass and hand speed in the performance of kung fu athletes compared with nonpractitioners

The main goal of this article is to quantify the contribution of effective mass (me) and handspeed (shi) on the palm strike performance of kung fu athletes (kung fu group) compared with nonpractitioners (control group). All subjects were asked to strike a basketball. Hand and ball speed (sbf) were determined by high-speed video analysis. The value for me was determined by an equation that does not depend upon postimpact measurement of the hand speed. The results show that kung fu athletes had greater shi (6.67 [SD 1.42] m/s), p = 0.042), higher me (2.62 [SD 0.33] kg, p = 0.004), and greater sbf (9.00 [SD 1.89] m/s), p = 0.004) than the nonpractitioners (5.04 [SD 0.57] m/s, 1.33 [SD 0.19] kg, and 5.72 [SD 0.44] m/s, respectively). Comparing the average values obtained for me and hand and forearm mass (m), it was found that for the control group me is statistically equal to m (p = 0.917), whereas for the kung fu group me is significant greater than m (p = 0.003). It is suggested that for impacts against heavier objects, the effective mass would be the main factor to distinguish a martial arts-trained from an untrained subject.     

Pathogenomics analysis of Leishmania spp.: flagellar gene families of putative virulence factors

The trypanosomatid flagellar apparatus contains conventional and unique features, whose roles in infectivity are still enigmatic. Although the flagellum and the flagellar pocket are critical organelles responsible for all vesicular trafficking between the cytoplasm and cell surface, still very little is known about their roles in pathogenesis and how molecules get to and from the flagellar pocket. The ongoing analysis of the genome sequences and proteome profiles of Leishmania major and L infantum, Trypanosoma cruzi, T. brucei, and T. gambiensi ( www.genedb.org ), coupled with our own work on L. chagasi (as part of the Brazilian Northeast Genome Program- www.progene.ufpe.br ), prompted us to scrutinize flagellar genes and proteins of Leishmania spp. promastigotes that could be virulence factors in leishmaniasis. We have identified some overlooked parasite factors such as the MNUDC-1 (a protein involved in nuclear development and genomic fusion) and SQS (an enzyme of sterol biosynthesis), among the described flagellar gene families. A database concerning the results of this work, as well as of other studies of Leishmania and its organelles, is available at http://nugen.lcc.uece.br/LPGate . It will serve as a convenient bioinformatics resource on genomics and pathology of the etiological agents of leishmaniasis.     

Alien barnacle Amphibalanus amphitrite epizoic on two native oyster species in the southern Gulf of Mexico: spatio-temporal variability and current status of its epibiosis

The study examines the current invasive status of the barnacle Amphibalanus amphitrite on natural populations of two native oyster species (the eastern oyster Crassostrea virginica and the mangrove oyster C. rhizophorae) in a estuary of the southern Gulf of Mexico. The spatio-temporal variability of the frequency of occurrence, density and coverage of A. amphitrite on shells of each oyster species was examined and compared with those of other native congeners (A. eburneus, A. venustus and A. improvisus) with which it coexists. These three variables were significantly higher in C. rhizophorae than in C. virginica and were within the range recorded for its congeners. The density of this invasive barnacle varied significantly at spatial and temporal scales. Its highest overall mean density occurred in February (at temperatures 26–27°C and salinities 29–32). Multivariate analyses showed a separation of Amphibalanus assemblages into three main groups according to variations in their composition and abundance among sampling sites and dates. It was also determined that salinity and sedimentation rate were the variables that best explain the distribution of the Amphibalanus species within the estuary. The limited presence of A. amphitrite on C. virginica reefs is likely due to (amongst other factors) the relatively higher sedimentation loads (370–5709?g?m^?2?d^?1) and lower salinities (9–23) recorded in these sites. This study can act as a reference point for the subsequent monitoring of this invasive species, since possible increases in its population density and competitive ability could threaten the integrity of oyster populations and other organisms in the southern Gulf of Mexico.     

EVOLUTION AND POPULATION STRUCTURE OF AFRICANIZED HONEY BEES IN BRAZIL: EVIDENCE FROM SPATIAL ANALYSIS OF MORPHOMETRIC DATA

In recent years, studies based on isoenzymatic patterns of geographic variation have revealed that what is usually called the Africanized honey bee does not constitute a single population. Instead, several local populations exist with various degrees of admixture with European honey bees. In this paper, we evaluated new data on morphometric patterns of Africanized honey bees collected at 42 localities in Brazil, using univariate and multivariate (canonical) trend surface and spatial autocorrelation analyses. The clinal patterns of variation found for genetically independent characters (wing size characters and some wing venation angles) are concordant with previous studies of malate dehydrogenase (MDH) allelic frequencies and support the hypothesis that larger honey bees in southern and southeastern Brazil originated by racial admixture in the initial phases of African honey bee colonization. Geographic variation patterns of Africanized honey bee populations reflect a demic diffusion process in which European genes were gradually lost because of the higher fitness of the African gene pool in Neotropical environmental conditions.