The multiple forms of bovine seminal ribonuclease: Structure and stability of a C-terminal swapped dimer

Bovine seminal ribonuclease (BS-RNase) acquires an interesting anti-tumor activity associated with the swapping on the N-terminal. The first direct experimental evidence on the formation of a C-terminal swapped dimer (C-dimer) obtained from the monomeric derivative of BS-RNase, although under non-native conditions, is here reported. The X-ray model of this dimer reveals a quaternary structure different from that of the C-dimer of RNase A, due to the presence of three mutations in the hinge peptide 111–116. The mutations increase the hinge peptide flexibility and decrease the stability of the C-dimer against dissociation. The biological implications of the structural data are also discussed.     

Molecular dynamics study of the structural basis of dysfunction and the modulation of reactive oxygen species generation by pathogenic mutants of human dihydrolipoamide dehydrogenase

Human dihydrolipoamide dehydrogenase (LADH, E3) is a component in the pyruvate-, alpha-ketoglutarate- and branched-chain ketoacid dehydrogenase complexes and in the glycine cleavage system. The pathogenic mutations of LADH cause severe metabolic disturbances, called E3 deficiency that often involve cardiological and neurological symptoms and premature death. Our laboratory has recently shown that some of the known pathogenic mutations augment the reactive oxygen species (ROS) generation capacity of LADH, which may contribute to the clinical presentations. A recent report concluded that elevated oxidative stress generated by the above mutants turns the lipoic acid cofactor on the E2 subunits dysfunctional. In the present contribution we generated by molecular dynamics (MD) simulation the conformation of LADH that is proposed to be compatible with ROS generation. We propose here for the first time the structural changes, which are likely to turn the physiological LADH conformation to its ROS-generating conformation. We also created nine of the pathogenic mutants of the ROS-generating conformation and again used MD simulation to detect structural changes that the mutations induced in this LADH conformation. We propose the structural changes that may lead to the modulation in ROS generation of LADH by the pathogenic mutations.     

Activation of Bacillus spores at moderately elevated temperatures (30–33 °C)

The time/temperature profiles experienced by spores on the track from their natural sporulation environment to consumable food products may be highly diverse. Temperature has been documented as an important factor that may activate spores, i.e. potentiates spores to germinate. There is, however, limited knowledge about the relationship between the expected temperature history and the subsequent germination characteristics of bacterial spores. We show here that the germination rate of five different Bacillus spore populations, represented by strains of Bacillus cereus, Bacillus weihenstephanensis, Bacillus pumilus, Bacillus licheniformis and Bacillus subtilis could be increased following 1 week storage at moderately elevated temperatures, 30–33 °C, compared to spores stored at 3–8 °C. The results imply that spores contamination routes to foods, specifically the temperature history, could be highly relevant data in predictive modeling of food spoilage and safety. Activation at these moderately elevated temperatures may be a native form of spore activation in their natural habitats, knowledge that also could be useful in development of decontamination strategies for mildly heated foods.     

Genetic differentiation of the Euglossini (Hymenoptera, Apidae) populations on a mainland coastal plain and an island in southeastern Brazil

Euglossini bees are among the main pollinators of plant species in tropical and subtropical forests in Central and South America. These bees are known as long-distance pollinators due to their exceptional flight performance. Here we assessed through microsatellite loci the gene variation and genetic differentiation between populations of four abundant Euglossini species populations sampled in two areas, Picinguaba (mainland) and Anchieta Island, Ubatuba, São Paulo State, southeastern Brazil. There was no significant genetic differentiation between the island and mainland samples of Euglossa cordata (Fst = 0.008, P = 0.60), Eulaema cingulata (Fst = 0.029, P = 0.29) and Eulaema nigrita (Fst = 0.062, P = 0.38), but a significant gene differentiation between mainland and island samples of Euglossa stellfeldi (Fst = 0.028, P = 0.016) was detected. As expected, our results showed that the water body that separates the island from the mainland does not constitute a geographic barrier for these Euglossini bees. The absence of populational structuring of three out the four species studied corroborates previous reports on those bees, characterized by large populations, with high gene diversity and gene flow and very low levels of diploid males. But the Eg. stellfeldi results clearly point that dispersal ability is not similar to all euglossine bees, what requires the development of different conservationist strategies to the Euglossini species.     

Pollen Harvest by Stingless Bee Foragers (Hymenoptera,Apidae, Meliponinae)

Data on pollen load capacity and flower constancy are discussed for nine stingless bee species. The foragers present high levels of flower constancy and often visit only one flower type (on average 97% of bee foragers), rarely a few flower types (on average 3% of bee foragers), during the same foraging trip. The latter foragers exhibit no tendencies for choosing similar sources, related either to flower type or to pollen type. Pollen load capacity (the ratio between pollen load weight/worker weight) decreases as forager body weight increases, so it is larger in smaller stingless bees species and smaller in larger ones. Nevertheless, it seems that specific load capacity also depends on pollen types.     

Microbial biofilm structure and organic matter use in mediterranean streams

River and stream biofilms in mediterranean fluvial ecosystems face both extreme seasonality as well as arrhythmic fluctuations. The hydrological extremes (droughts and floods) impose direct changes in water availability but also in the quantity and quality of organic matter and nutrients that sustain the microbial growth. This review analyzes how these ecological pulses might determine unique properties of biofilms developing in mediterranean streams. The paper brings together data from heterotrophic and autotrophic community structure, and extracellular enzyme activities in biofilms in mediterranean streams. Mediterranean stream biofilms show higher use of peptides during the favorable period for epilithic algae development (spring), and preferential use of cellulose and hemicellulose in autumn as a response to allochthonous input. The drying process causes the reduction in bacterial production and chlorophyll biomass, but the rapid recovery of both autotrophs and heterotrophs with rewetting indicates their adaptability to fluctuations. Bacteria surviving the drought are mainly associated with sediment and leaf litter which serve as “humid refuges”. Some algae and cyanobacteria show resistant strategies to cope with the drought stress. The resistance to these fluctuations is strongly linked to the streambed characteristics (e.g., sediment grain size, organic matter accumulation, nutrient content).     

Cyanobacterial dominance in Brazil: distribution and environmental preferences

Based on a literature survey, we evaluated the periods of cyanobacterial dominance in Brazil. We hypothesized that variability of environmental forces along the country will promote or facilitate temporal and spatial mosaic in cyanobacterial dominance. The most striking outcomes are related to the dominance of Cylindrospermopsis, Dolichospermum, and Microcystis. Although they share important adaptive strategies (e.g., aerotopes, large size and toxins production), our findings suggest that they have different environmental preferences. Dolichospermum and Microcystis dominated mainly in warm-rainy periods whereas Cylindrospermopsis was more common during dry periods and in mixed systems, or formed perennial dominance. Maximum phosphorus concentrations were observed in reservoirs dominated by Cylindrospermopsis. Although the main genera reached high biomass levels individually, different abilities to form dominance and co-dominance were observed. The number of co-dominance of Chroococales and Nostocales was almost the same as the individual occurrence of the main genera from these groups. This dataset reveals patterns of dominance of these cyanobacteria and also indicates that physiological features will cause differences in the mechanisms of interactions between species. The understanding of these processes and their relationship to environmental conditions will promote better understanding of cyanobacterial dominance and increase our ability to predict and manage these events.     

Abstracts of papers read at the Winter Meeting, 1961

Herdmania litoralis is a heterotrophic, sand-dwelling dinoflagellate with morphological characters that do not provide clear evidence for its systematic position in any existing family of dinoflagellates. Protoperidinium minutum is a heterotrophic, planktonic species that has a typical tabulation for the genus Protoperidinium. In order to infer the phylogenetic positions of these two species more confidently, we characterized the thecal plate patterns and determined small-subunit and large-subunit ribosomal DNA sequences (SSU rDNA and LSU rDNA, respectively) from both species. Intraindividual and intraspecific diversity of SSU and LSU rDNA data were characterized in H. litoralis using a combination of single-cell PCR approaches and analyses of PCR clones derived from multi-cell DNA extractions. The results of the molecular phylogenetic analyses demonstrated a novel, well-supported clade comprising both sand-dwelling species (H. litoralis and Thecadinium dragescoi) and planktonic species (P. minutum). Because the establishment of this clade also demonstrated that P. minutum is not a member of Protoperidinium, we reinstated and emended the genus Archaeperidinium Jörgensen 1912 Jörgensen, E. 1912. Bericht über die von der schwedischen hydrographisch-biologischen Kommission in den schwedischen Gewässern in den Jahren 1909–10 eingesammelten Planktonproben. Svenska Hydrograph.-Biol. Komm. Skr., 4: 1–20.  [Google Scholar].     

Synthesis and cytotoxic activity of metallic complexes of lawsone

In the present study, a series of metallic complexes of the 1,4-naphthoquinone lawsone (2–6) were synthesized and evaluated for potential cytotoxicity in a mouse leukemic macrophagic RAW 264.7 cell line. Cell viability was determined by the MTT assay. Significant growth inhibition was observed for the copper complex (4) with an IC₅₀ value of 2.5 μM. This compound was selected for further evaluation of cytotoxic activity on several human cancer cells including HT-29 (human colorectal adenocarcinoma), HepG2 (human hepatocellular carcinoma) and HeLa, (human cervical adenocarcinoma cells). Significant cell viability decrease was also observed in HepG2 cells. The apoptotic potential of this complex was evaluated in these cells. Compound 4 induced apoptosis by a mechanism that involves the activation of caspases 3, 8 and 9 and modulation of apoptotic-related proteins such as Bax, Bad, and p53. These results indicate that metal complexes of lawsone derivatives, in particular compound 4, might be used for the design of new antitumoral agents.