Polyphosphate accumulation and cell-surface properties by autochthonous bacteria from Argentinian Patagonia.

Bacteria persisting in environments contaminated with polycyclic aromatic hydrocarbons (PAHs) have developed physiological mechanisms to counteract environmental stress. Inorganic polyphosphate accumulation represents one of these possible mechanisms. Likewise, properties such as cell-surface hydrophobicity, auto-aggregation, biofilm formation and bioemulsifying activity could facilitate interaction of microorganisms with hydrophobic organic compounds. In this work, these physiological properties were compared in indigenous bacteria from polluted sediments from Argentinian Patagonia, which were cultivated in two culture media (LBm and JPP) as a way to improve in the next future the PAHs removal. The highest hydrophobicity values were obtained in Rhodococcus strains, while Bacillus sp. B18 showed the highest auto-aggregation percentage and emulsion index. The highest numerical values of biofilm formation were determined in Rhodococcus sp. F27, Pseudomonas sp. P26, and Gordonia sp. H19 either on hydrophilic or on hydrophobic support. The qualitative and quantitative polyP determinations confirmed the presence of this biopolymer in the strains evaluated. The highest intracellular phosphate mean values were obtained in Bacillus sp. B18 in LBm and Rhodococcus erythropolis 20 in JPP. The bacteria evaluated belonging to different genera showed significant differences in their cell-surface characteristics, bioemulsifying activity and polyP accumulation. The low-cost JPP culture medium was selected for future contaminant removal studies.

Petroleum oil removal by immobilized bacterial cells on polyurethane foam under different temperature conditions.

In this work, a mixed biofilm composed by Pseudomonas monteilii P26 and Gordonia sp. H19 was formed using polyurethane foam (PUF) as immobilization support, for crude oil removal from artificial sea water. Fresh immobilized cells and immobilized cells that were stored at 4°C for two months before use were assessed. The oil removal assays were carried out at microcosm scale at 4, 15 and 30°C. A viability loss of P. monteilii P26 was observed after the storage. The highest removal value (75%) was obtained at 30°C after 7days using fresh immobilized cells on PUF. Enhanced oil bioremoval was obtained at 4°C and 15°C with the previously stored immobilized cells compared to the fresh immobilized cells. Crude oil sorption on the different systems was responsible for the removal of 22-33% oil at the different temperatures. In conclusion, an economic tool for petroleum bioremediation is proposed.

Insight into the Binding of DFG-out Allosteric Inhibitors to B-Raf Kinase Using Molecular Dynamics and Free Energy Calculations.

B-Raf mutations are identified in 40-50% of patients with melanoma and among them, the substitution of valine for glutamic acid at position 600 ((V600E)B-Raf) is the most frequent. Treatment of these patients with B-Raf inhibitors has been associated with a clear clinical benefit. Unfortunately, multiple resistance mechanisms have been identified and new potent and selective inhibitors are currently needed. In this work, five different type II inhibitors, which bind (V600E)B-Raf in its DFG-out conformation, have been studied using molecular dynamics, free energy calculations and energy decomposition analysis. The ranking of calculated MM-PB/GBSA binding affinities is in good agreement with the experimentally measured ones. The per-residue decomposition of ΔGbinding, within the MM-GBSA approach, has been used to identify the key residues governing the allosteric binding of the studied compounds to the (V600E)B-Raf protein kinase. Results indicate that although van der Waals interactions are key determinants for binding, hydrogen bonds also play an important role. This work also provides a better structural understanding of the binding of DFG-out inhibitors to (V600E)B-Raf, which can be used in a further step for rational design of a new class of B-Raf potent inhibitors.

Urogenital pathogen inhibition and compatibility between vaginal Lactobacillus strains to be considered as probiotic candidates.

OBJECTIVES: To evaluate the production of inhibitory substances against several urogenital pathogens and to determine the compatibility between selected vaginal lactobacilli to be considered as probiotic candidates. STUDY DESIGN: The inhibitory activity of 38 vaginal Lactobacillus strains against urogenital pathogens was assessed using the agar plate diffusion. METHOD: The production of hydrogen peroxide was determined by the semi-qualitative tetramethyl-benzidine-peroxidase assay and that of lactic acid by high performance liquid chromatography. Pre-selected lactobacilli were genetically identified by ribosomal 16S-DNA sequencing and used to determine the compatibility of strains. Correlation values between some inhibitory properties were calculated. RESULTS: 45% of the lactobacilli assayed inhibited the growth of various urogenital pathogens, except Candida albicans, mainly by the effect of organic acids. Even though 71% of the Lactobacillus strains produced hydrogen peroxide in the semi-qualitative test, the antimicrobial effect of the oxidative metabolite on pathogens was not detected under the experimental conditions assayed. On the other hand, the inhibition spectrum of salivaricin CRL 1328 was widened with respect to the one previously reported by our research team. With respect to the degree of interaction between the pre-selected lactobacilli, most of them showed inhibition of at least one strain, mainly due to the effects of both organic acids and hydrogen peroxide. CONCLUSION: Several lactobacilli that showed inhibitory activity against urogenital pathogens or produced hydrogen peroxide are compatible between them. Therefore, different potential combinations of strains are proposed for their inclusion in the design of probiotic products to be vaginally administered for the prevention of urogenital infections in women.