Biochemical and structural characterization of Salmonella typhimurium glyoxalase II: new insights into metal ion selectivity

Glyoxalase II is a hydrolytic enzyme part of the glyoxalase system, responsible for detoxifying several cytotoxic compounds employing glutathione. Glyoxalase II belongs to the superfamily of metallo-beta-lactamases, with a conserved motif able to bind up to two metal ions in their active sites, generally zinc. Instead, several eukaryotic glyoxalases II have been characterized with different ratios of iron, zinc, and manganese ions. We have expressed a gene coding for a putative member of this enzyme superfamily from Salmonella typhimurium that we demonstrate, on the basis of its activity, to be a glyoxalase II, named GloB. Recombinant GloB expressed in Escherichia coli was purified with variable amounts of iron, zinc, and manganese. All forms display similar activities, as can be shown from protein expression in minimal medium supplemented with specific metal ions. The crystal structure of GloB solved at 1.4 A shows a protein fold and active site similar to those of its eukaryotic homologues. NMR and EPR experiments also reveal a conserved electronic structure at the metal site. GloB is therefore able to accommodate these different metal ions and to carry out the hydrolytic reaction with similar efficiencies in all cases. The metal promiscuity of this enzyme (in contrast to other members of the same superfamily) can be accounted for by the presence of a conserved Asp residue acting as a second-shell ligand that is expected to increase the hardness of the metal binding site, therefore favoring iron uptake in glyoxalases II.     

Effect of fertilization levels, age and canopy height of Lycopersicon hirsutum on the resistance to Myzus persicae

The effects of different levels of fertilization with nitrogen and potassium (NK), height within the canopy and plant age of Lycopersicon hirsutum f. glabratum (PI 134417) C. H. Mull and L. esculentum on the density of foliar cristalliferous idioblasts and trichomes, and on the levels of 2- tridecanone (2-TD) and 2-undecanone (2-UD) and their influence on the resistance to Myzus persicae (Sulzer) were investigated. NK levels were inversely related to densities of trichomes and cristalliferous idioblasts in L. esculentum, but only to densities of cristalliferous idioblasts in L. hirsutum. Trichome density increased with increasing height within the canopy and with plant age in L. esculentum and L. hirsutum. Density of cristalliferous idioblasts in L. hirsutum increased with canopy height reaching a maximum at mid-height within the canopy in opposition to L. esculentum in which a small decrease in density with height within the canopy was found. There was also a small decrease of idioblast density with increasing plant age for L. esculentum, while a greater and opposite variation took place in L. hirsutum. L. esculentum has mainly non- glandular leaf trichomes (90%), while L. hirsutum has mainly glandular trichomes (97%). The main glandular trichomes of L. hirsutum are producers of 2-TD and 2-UD, whose leaf content increases with increase of trichome density. A negative effect of 2-TD concentration and cristalliferous idioblast density on the survival and longevity of M. persicae was observed. Since L. hirsutum presented much higher levels of 2-TD and cristalliferous idioblast density than L. esculentum, L. hirsutum seems a promising source of resistance to M. persicae for tomato breeding programs.     

Hierarchical Bayesian Estimation for the Number of Species

This paper is concerned with the estimation of the number of species in a population through a fully hierarchical Bayesian model using the Metropolis algorithm. The proposed Bayesian estimator is based on Poisson random variables with means that are distributed according to some prior distributions with unknown hyperparameters. An empirical Bayes approach is considered and compared with the fully Bayesian approach based on biological data.     

Assessment of mite fauna in different coffee cropping systems in Brazil

Agro-ecological diversification may increase natural biodiversity and decrease environmental impacts caused by the intensification of management practices, especially with respect to pest control. This study aimed to evaluate mite populations from surveys in commercial coffee (Coffea arabica) orchards managed in different ways. Four different cropping systems were compared: a conventional system (CS), non-pesticide system (NPS), organic system (OS) and organic agroforestry system (OAS). Mite samples were taken from 24 coffee plantations (n?=?6 for each management strategy). Six leaves per plant were collected from five sampling sites totalling 30 leaves from each plantation. Mites were removed from individual leaves, counted and identified to family level. Amongst the predatory mites, two families were detected; the Phytoseiidae was the most abundant family while the Stigmaeidae, generalist predatory mites, were only found in two management systems (NPS and AOS). Of the two families of phytophagous mites, the family Tenuipalpidae was the most abundant, but rarely found in AOS. The other phytophagous family, Tetranychidae, was less abundant in coffee crops than Tenuipalpidae, but the most abundant phytophagous family in CS. Understanding the relationship between mite populations and management practices can aid in choice of spraying strategies for different coffee management systems.     

Trophic role of demersal mesopredators on rocky reefs in an equatorial Atlantic Ocean island

The decrease in the number of sharks around Saint Peter and Saint Paul Archipelago (SPSPA) may impact food web structure. We investigated trophic relationships in the shallow rocky reefs of the SPSPA using stable isotopes and stomach contents with a particular focus on three abundant mesopredators: Caranx lugubris, Enchelycore nigricans and Muraena pavonina. Food web structure was described using samples of the most abundant basal resources, fishes and invertebrates, which were collected in April and October 2012. Individuals of the three focal species (n = 138: C. lugubris, n = 56; E. nigricans, n = 18; M. pavonina, n = 64) were collected during four expeditions to SPSPA (April and October of 2011 and 2012). Results suggest that this shallow water food web is supported by trophic pathways originating from benthic resources. Stable isotope data suggest potential competitive interactions between the whitespot moray and the other two mesopredators. Conversely, stomach content data suggest little niche overlap in the three focal species, but these data must be interpreted carefully because of the small sample sizes and restricted temporal sampling windows. All three mesopredators have a significant, albeit weak, relationship between body size and δ¹⁵N, suggesting ontogenetic diet shifts. These data contribute baseline information to assess shifts in food web structure that may stem from top predator decline in this unique ecosystem.     

In Silico,In Vitro and In Vivo Toxicological Assessment of BPP-BrachyNH<Subscript>2</Subscript>, A Vasoactive Proline-Rich Oligopeptide from <Emphasis Type="Italic">Brachycephalus ephippium</Emphasis>

BPP-BrachyNH₂ is a proline-rich oligopeptide (PRO) firstly identified in skin secretion of the frog Brachycephalus ephippium, which possess in vitro inhibitory activity of angiotensin-I converting enzyme (ACE) and endothelium-dependent vasorelaxant activity. Considering its potential application in the treatment of cardiovascular diseases, the present work assessed the toxicological profile of the BPP-BrachyNH₂. The in silico toxicity prediction was performed from the best model obtained through the optimization of the FASTA query peptide. This prediction study revealed that BPP-BrachyNH₂ induced high predicted LD₅₀ values for both humans and rats, and then is well-tolerated in the recommended range. The MTT assay was applied for the in vitro cytotoxic evaluation in murine macrophages. In this assay, a decrease of cell viability was not observed. The in vivo acute toxicological study was performed after the intraperitoneal administration of BPP-BrachyNH₂ at doses of 5 and 50 mg/kg. After intraperitoneal administration, no death, alterations in behavioral parameters or weight gain curve was observed, as well as none in the serum biochemical parameters, and gross pathological and histopathological analyses. These observations demonstrates an acceptable safety profile for BPP-BrachyNH₂, leading towards further studies focused on investigation of pharmacological and therapeutical applications for this peptide.     

Dermaseptins from Phyllomedusa oreades and Phyllomedusa distincta: liposomes fusion and/or lysis investigated by fluorescence and atomic force microscopy

Three dermaseptins, DS 01, DD K, and DD L, were compared with respect to their structural features and interactions with liposomes. Circular dichroic spectra at alcohols of different chain lengths revealed that DS 01 has the higher helicogenic potential in hydrophobic media. Binding of DS 01, DD K, and DD L to liposomes induced significant blue shifts of the emission spectra of the single tryptophan located at position 3 of all sequences indicating association of the peptides with bilayers. Kinetics evaluation of atomic force microscopy images evidenced the strong fusogenic activity of DS 01 whereas DD K and DD L showed increased lytic activities.     

Neonatal immunization with a single dose of recombinant BCG expressing subunit S1 from pertussis toxin induces complete protection against Bordetella pertussis intracerebral challenge

The currently used pertussis vaccines are highly efficacious; however, neonates are susceptible to whooping cough up to the sixth month. In agreement, DTP-immunized neonate mice were not protected against intracerebral challenge with Bordetella pertussis. Neonate mice immunized with either DTP or a recombinant-BCG strain expressing the genetically detoxified S1 subunit of pertussis toxin do not show a humoral immune response against PT. On the other hand, rBCG-Pertussis induces higher PT-specific IFN-gamma production and an increase in both IFN-gamma(+) and TNF-alpha(+)-CD4(+)-T cells than the whole cell pertussis vaccine and confers protection against a lethal intracerebral challenge with B. pertussis.