Toxicological effect and enzymatic disorder of non-studied emerging contaminants in Artemia salina model.

Emerging contaminants such as sunscreens, hair dyes and flame retardants have been found at important concentrations in surface water (river, lake, ocean), but their negative impact on different aquatic species is not fully known. This study evaluated the effect of benzophenone (BZ), 2,5-diaminotoluene sulfate (PTD), p-phenylenediamine (PPD) and tetrabromobisphenol A (TBPA) on survival (LC50) and the impact of sublethal concentrations (LC25) on the activity of enzymes linked to stress oxidative process in brine shrimp under two temperature conditions (22 °C and 28 °C) for 24 h and 48 h of exposure time. LC50 values obtained for each chemical substance and the activity of GST, AChE and LDH were significantly affected by the temperature conditions and exposure time. In contrast, GPx was only altered by the tested compound. TBBPA (LC50 from 17.05 up to 28.55 µg/L) and BZ (LC50 from 14.86 up to 24.49 mg/L) resulted in the most toxic substances for A. salina. The impact of dyes, such as PTD and PPD, on aquatic organisms is limited. These are the first results that show that not only dyes, but their respective by-products induce harmful effects in brine shrimp (LC50 for PTD and PPD were 23.6-396.3 and 52.0-164.9 mg/L respectively). Although this study model was very useful to evaluate the ecotoxicity of the different ECs, additional research is needed to increase available information related to the effects of dyes and other non-studied micropollutants on aquatic systems in general.

Discrimination of radiosensitive and radioresistant murine lymphoma cells by Raman spectroscopy and SERS.

Intrinsic radiosensitivity is a biological parameter known to influence the response to radiation therapy in cancer treatment. In this study, Raman spectroscopy and surface enhanced Raman spectroscopy (SERS) were successfully used in conjunction with principal component analysis (PCA) to discriminate between radioresistant (LY-R) and radiosensitive (LY-S) murine lymphoma sublines (L5178Y). PCA results for normal Raman analysis showed a differentiation between the radioresistant and radiosensitive cell lines based on their specific spectral fingerprint. In the case of SERS with gold nanoparticles (AuNPs), greater spectral enhancements were observed in the radioresistant subline in comparison to its radiosensitive counterpart, suggesting that each subline displays different interaction with AuNPs. Our results indicate that spectroscopic and chemometric techniques could be used as complementary tools for the prediction of intrinsic radiosensitivity of lymphoma samples.

Oxidation of Flame Retardant Tetrabromobisphenol A by a Biocatalytic Nanofiber of Chloroperoxidase.

BACKGROUND: Tetrabromobisphenol (TBBPA), a flame retardant compound, is considered a ubiquitous pollutant, with potential impact on the environment and human health. Several technologies have been applied to accelerate its degradation and minimize environmental impacts. Due to its aromaticity character, peroxidase enzymes may be employed to carry out its transformation in mild conditions. Therefore, the purpose of this work was to determine the capacity of the enzyme chloroperoxidase (CPO) to oxidize TBBPA in several water samples. METHODS: The oxidation capacity of CPO was evaluated in catalytic conditions using water samples from surface and groundwater, as well as effluents from wastewater treatment plants. The biocatalytic performance of CPO was improved due to its immobilization on nanofibers composed of polyvinyl alcohol and chitosan (PVA/chitosan). RESULTS: Free and immobilized CPO were able to transform more than 80% in short reaction times (60 min); producing more biodegradable and less toxic products. Particularly, the immobilized enzyme was catalytically active in a wider range of pH than the free enzyme with the possibility of reusing it up to five times. CONCLUSIONS: The biocatalytic oxidation of TBBPA under environmental conditions is highly efficient, even in complex media such as treated effluents of wastewater treatment plants.

Transcription factor TFIIEß interacts with two exposed positions in helix 2 of the Antennapedia homeodomain to control homeotic function in Drosophila.

Homeoproteins contain the conserved homeodomain (HD) and have an important role determining embryo body plan during development. HDs increase their DNA-binding specificity by interacting with additional cofactors outlining a Hox interactome with a multiplicity of protein-protein interactions. In Drosophila, the first link of functional contact with a general transcription factor (GTF) was found between Antennapedia (Antp) and BIP2 (TFIID complex). Hox proteins also interact with other components of Pol II machinery such as the subunit Med19 from Mediator (MED) complex, TFIIEß and transcription-pausing factor M1BP. All these interactions clearly demonstrate Hox-driven transcriptional regulation, but the precise molecular mechanism remains unclear. In this paper, we focused on the Antp-TFIIEß protein-protein interface to establish the specific contacts as well as its functional role. Using Bimolecular Fluorescence Complementation (BiFC) in cell culture and in vivo we found that TFIIEß interacts with Antp through the HD independently of the YPWM motif and the direct physical interaction is at helix 2, specifically aminoacidic positions I32 and H36 of Antp. We also found, through ectopic assays, that these two positions in helix 2 are crucial for Antp homeotic function in head involution, and thoracic and antenna-to tarsus transformations. Interestingly, overexpression of Antp and TFIIEß in the antennal disc showed that this interaction is required for the antenna-to-tarsus transformation. In conclusion, interaction of Antp with TFIIEß is important for the functional specificity of Antennapedia, and amino acids 32 and 36 in Antp HD helix 2 are key for this interaction. Our results open the possibility to more broadly analyze Antp-TFIIEß interaction on the transcriptional control for the activation and/or repression of target genes in the Hox interactome during Drosophila development.

Extraction and purification of high-value metabolites from microalgae: essential lipids, astaxanthin and phycobiliproteins.

The marked trend and consumers growing interest in natural and healthy products have forced researches and industry to develop novel products with functional ingredients. Microalgae have been recognized as source of functional ingredients with positive health effects since these microorganisms produce polyunsaturated fatty acids, polysaccharides, natural pigments, essential minerals, vitamins, enzymes and bioactive peptides. For this reason, the manuscript reviews two of the main high-value metabolites which can be obtained from microalgae: pigments and essential lipids. Therefore, the extraction and purification methods for polyunsaturated fatty acids, astaxanthin, phycoerythrin and phycocyanin are described. Also, the effect that environmental growth conditions have in the production of these metabolites is described. This review summarizes the existing methods to extract and purify such metabolites in order to develop a feasible and sustainable algae industry.

Enhanced production of thermostable laccases from a native strain of Pycnoporus sanguineus using central composite design.

The production of thermostable laccases from a native strain of the white-rot fungus Pycnoporus sanguineus isolated in Mexico was enhanced by testing different media and a combination of inducers including copper sulfate (CuSO4). The best conditions obtained from screening experiments in shaken flasks using tomato juice, CuSO4, and soybean oil were integrated in an experimental design. Enhanced levels of tomato juice as the medium, CuSO4 and soybean oil as inducers (36.8% (v/v), 3 mmol/L, and 1% (v/v), respectively) were determined for 10 L stirred tank bioreactor runs. This combination resulted in laccase titer of 143,000 IU/L (2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), pH 3.0), which represents the highest activity so far reported for P. sanguineus in a 10-L fermentor. Other interesting media resulting from the screening included glucose-bactopeptone which increased laccase activity up to 20,000 IU/L, whereas the inducers Acid Blue 62 and Reactive Blue 19 enhanced enzyme production in this medium 10 times. Based on a partial characterization, the laccases of this strain are especially promising in terms of thermostability (half-life of 6.1 h at 60 °C) and activity titers.

Integrating toxin gene expression, growth and fumonisin B1 and B2 production by a strain of Fusarium verticillioides under different environmental factors.

The objective of this study was to integrate data on the effect of water activity (a(w); 0.995-0.93) and temperature (20-35 °C) on activation of the biosynthetic FUM genes, growth and the mycotoxins fumonisin (FB1, FB2) by Fusarium verticillioides in vitro. The relative expression of nine biosynthetic cluster genes (FUM1, FUM7, FUM10, FUM11, FUM12, FUM13, FUM14, FUM16 and FUM19) in relation to the environmental factors was determined using a microarray analysis. The expression was related to growth and phenotypic FB1 and FB2 production. These data were used to develop a mixed-growth-associated product formation model and link this to a linear combination of the expression data for the nine genes. The model was then validated by examining datasets outside the model fitting conditions used (35 °C). The relationship between the key gene (FUM1) and other genes in the cluster (FUM11, FUM13, FUM9, FUM14) were examined in relation to aw, temperature, FB1 and FB2 production by developing ternary diagrams of relative expression. This model is important in developing an integrated systems approach to develop prevention strategies to control fumonisin biosynthesis in staple food commodities and could also be used to predict the potential impact that climate change factors may have on toxin production.

Functional synthetic Antennapedia genes and the dual roles of YPWM motif and linker size in transcriptional activation and repression.

Segmental identity along the anteroposterior axis of bilateral animals is specified by Hox genes. These genes encode transcription factors, harboring the conserved homeodomain and, generally, a YPWM motif, which binds Hox cofactors and increases Hox transcriptional specificity in vivo. Here we derive synthetic Drosophila Antennapedia genes, consisting only of the YPWM motif and homeodomain, and investigate their functional role throughout development. Synthetic peptides and full-length Antennapedia proteins cause head-to-thorax transformations in the embryo, as well as antenna-to-tarsus and eye-to-wing transformations in the adult, thus converting the entire head to a mesothorax. This conversion is achieved by repression of genes required for head and antennal development and ectopic activation of genes promoting thoracic and tarsal fates, respectively. Synthetic Antennapedia peptides bind DNA specifically and interact with Extradenticle and Bric-à-brac interacting protein 2 cofactors in vitro and ex vivo. Substitution of the YPWM motif by alanines abolishes Antennapedia homeotic function, whereas substitution of YPWM by the WRPW repressor motif, which binds the transcriptional corepressor Groucho, allows all proteins to act as repressors only. Finally, naturally occurring variations in the size of the linker between the homeodomain and YPWM motif enhance Antennapedia repressive or activating efficiency, emphasizing the importance of linker size, rather than sequence, for specificity. Our results clearly show that synthetic Antennapedia genes are functional in vivo and therefore provide powerful tools for synthetic biology. Moreover, the YPWM motif is necessary--whereas the entire N terminus of the protein is dispensable--for Antennapedia homeotic function, indicating its dual role in transcriptional activation and repression by recruiting either coactivators or corepressors.