ECOTOX - biomonitoring based on real time movement analysis of unicellular organisms

To meet the todays needs for the protection of the environment from pollution through cumulative poisonings or biohazards ecotoxicology uses biotests, to determine effects of chemicals and sewage waters to ecosystems. ECOTOX is a biotest system that allows both the estimation of risks arising from certain substances or substance mixtures as well as the on-line monitoring of waste waters and aquatic ecosystems. Euglena gracilis, the employed organism for freshwater measurements, found to be highly sensitive to external factors, provides several physiological endpoints, well fitted for toxicity hazard assessment in water management, which can be characterized by the ECOTOX program.     

Roles of superoxide dismutase and catalase of Staphylococcus xylosus in the inhibition of linoleic acid oxidation

Streptomyces brasiliensis ATCC 23727 showed extensive sporulation when cultured in a liquid medium containing galactose and glutamic acid as carbon and nitrogen sources. Under such conditions, glycogen and trehalose are accumulated in the hyphae coinciding with spore formation. The results reported here suggest that glycogen accumulated in sporogenic hyphae is converted into trehalose during the final period of spore maturation. Glycogen is also accumulated in the hyphae when S. brasiliensis is cultured under conditions which did not support sporulation. Under such conditions, however, glycogen degradation is not accompanied by accumulation of trehalose. This suggest that the conversion of glycogen into trehalose might be a sporulation-specific event in streptomycetes.     

Synthetic ion transporters: Pore formation in bilayers via coupled activity of non-spanning cobalt-cage amphiphiles

Three amphiphilic cobalt-cage congeners bearing a diaza-crown bridge and varying alkyl chains (1:2:3; n = 12, 16, 18) have been assessed for their ion transport across planar lipid bilayer membranes. In symmetrical electrolyte solutions, a range of ion transport activity is provoked: 1 disrupts painted (fluid) bilayers in a detergent-like mode of action; 2 forms conducting “pores” in folded (rigid) membranes with long open lifetimes (> 2 min) while 3 requires the larger auxiliary solvent volume and lower lateral stress of painted membranes to effect ion transport via long-lived pores. Hill analysis of the conductance variation with monomer concentration yields coefficients (2:3; n = 2.3, 1.9) in support of dimeric (n = 2) membrane-active structures, for which the derived “pore” radii are correlated with charge-density of the transported cations and their affinity for the crown moiety. A toroidal-pore model is invoked to account for the flux of guest ions through planar bilayer membranes without a fast-diffusing intermediary or direct membrane-spanning structure.     

Possible involvement of the membrane potential in the gravitactic orientation of Euglena gracilis

Euglena gracilis, a unicellular photosynthetic flagellate, uses light and gravity as environmental hints to reach and stay in regions optimal for growth and reproduction. The current model of gravitaxis (the orientation with respect to the earth's gravitational field) is based on the specific density difference between cell body and medium. The resulting sedimentation of the cell body applies a force to the lower membrane. This force activates mechano-sensitive ion channels. The resulting ion flux changes the membrane potential, which in turn triggers reorientational movements of the trailing flagellum. One possibility for recording the predicted membrane potential changes during reorientation is the use of potential-sensitive dyes, such as Oxonol VI. The absorption changes of the dye indicating potential changes were recorded with a custom-made photometer, which allows a high precision measurement with a high temporal resolution. After a gravitactic stimulation, a short period of hyperpolarization was detected, followed by a massive depolarization of the cell. The membrane potential returned to initial values after a period of approximately 200 s. Parallel measurements of the precision of orientation and the membrane potential showed a close relationship between both phenomena. The obtained results support the current model of gravitaxis of Euglena gracilis.     

Photosynthetic performance of the Atlantic brown macroalgae, Cystoseira abies-marina, Dictyota dichotoma and Sargassum vulgare, measured in Gran Canaria on site

Photosynthetic performance was determined in three common Atlantic brown macroalgae, Cystoseira abies-marina, Dictyota dichotoma and Sargassum vulgare, in Gran Canaria, Canary Islands, on site. The photosynthetic quantum yield was measured with both a portable PAM instrument on site and a diving PAM under water in the habitat. In parallel, solar radiation was measured continuously above and under water by means of two three-channel dosimeters, ELDONET (Real Time Computer, M?hrendorf, Germany), in three wavelength ranges, UV-A, UV-B and PAR. The effective photosynthetic quantum yield decreased in all species in response to exposure to 15 min of solar radiation but recovered in the subsequent shade conditions within several hours. A 30-min exposure caused an even more profound photoinhibition from which the algae recovered only partially. Most of the effect was due to visible radiation, however, the UV wavelength range, and especially UV-B, considerably enhanced the decrease in photosynthetic quantum yield. In all species except Sargassum a significant photoinhibition was detected at their growth sites at high solar angles in the water column, measured with the diving PAM.     

Behavioral mutants of Euglena gracilis: functional and spectroscopic characterization

Three mutant strains of the phytoflagellate Euglena gracilis Z have been characterized in order to analyze the signal perception and signal transduction pathways involved in photo- and gravitaxis. Using the fluorescence of the chromophoric groups believed to be involved in photoperception (flavins and pterins) a method was developed for an in situ and in vivo detection of the paraxonemal body, the proposed location of the photoreceptor molecules. Two of the mutant strains, 1224-5/9f and 1224-5/1f, do not possess a stigma and also lack a paraxonemal body, as indicated by fluorescence measurements. The third strain, FB, has a small stigma, but only some cells contain a paraxonemal body. In contrast to the present hypothesis on photoorientation of Euglena, all strains were able to orient with respect to the light direction. However, the mutant strains did not show any orientation at low irradiances. At medium and high irradiances the strains 1224-5/9f and 1224-5/1f oriented perpendicular to the light direction (diaphototaxis) while cells of strains of FB showed partly negative phototaxis and partly diaphototaxis. Diaphototaxis was never observed in the wild type strain. Strains 1224-5/9f and 1224-5/1f showed normal graviresponses compared with the wild type. Astasia longa, a nonphtototactic relative of E. gracilis, as well as strain FB were both negative and positive gravitactic at all culture ages tested. This result confirmed the hypothesis that the paraxonemal body is not directly involved in graviperception.     

Physiological characterization of gravitaxis in Euglena gracilis

The motile, unicellular freshwater flagellate Euglena gracilis uses external stimuli, like gravity, light or oxygen pressure in order to orient itself in its natural habitat. In the darkness the cells normally show a negative gravitactic behavior, that means they swim upward in the water column, Many ground and space experiment revealed that gravitaxis is most likely based on active physiological mechanisms (involvement of calcium, cAMP, membrane potential and other parameters).     

Towards guidance values for the environmental performance of buildings: application to the statistical analysis of 40 low-energy single family houses’ LCA in France

Purpose

In this study, life cycle assessment (LCA) is applied to a sample of 40 low-energy individual houses for the French context in order to identify guidance values for different environmental priorities (energy and water consumption, greenhouse gases emissions, waste generation etc.).

Methods

Calculation rules for the LCA derived from EeBGuide guidance and HQE Performance specific rules for the French context. Data are based on Environmental Product Declaration (EPD for the impacts related to products and technical equipment while generic data are used for energy and water processes. The LCA is defined for the entire life cycle of a building from cradle-to-grave according to NF EN 15978 standard. It includes the products and equipment implemented in the building, the different uses of energy for heating, domestic hot water, lighting, ventilation and auxiliaries, and the different uses of water consumption.

Results and discussion

Results for the 40 houses showed that the average life cycle non-renewable primary energy consumption is about 37 kWh/(m²*year) while the life cycle greenhouse gases emissions are of 8.4 kg CO₂-eq/(m²*year). The embodied impacts represent between 40% and 72% for the following indicators: acidification, global warming, non-renewable primary energy, and radioactive waste. The net fresh water use is mostly determined by the direct use of the water in use, and the non-hazardous waste indicator is only linked to the materials and equipment. When integrating the variability of the different houses design, energy performance, climate requirements, it was found that those values can vary of an order of two between the 10 and 90% percentiles’ values. It was found that the results are also sensitive to the enlargement of the system boundaries (e.g. inclusion of the other uses of energy such as building appliances) and the modification of the reference study period.

Conclusions and recommendations

This study provided a first set of LCA guidance values describing a range of environmental impacts for new low-energy individual houses in France. Results were also reported for different design parameters, system boundaries and reference study period. The outcomes of this study can now serve as a basis to guide and support new LCA-based labelling systems developed by public authorities and labelling schemes (e.g. the HQE Association).

     

Development of a Protein Standard Absolute Quantification (PSAQ™) assay for the quantification of Staphylococcus aureus enterotoxin A in serum

Enterotoxin A (SEA) is a staphylococcal virulence factor which is suspected to worsen septic shock prognosis. However, the presence of SEA in the blood of sepsis patients has never been demonstrated. We have developed a mass spectrometry-based assay for the targeted and absolute quantification of SEA in serum. To enhance sensitivity and specificity, we combined an immunoaffinity-based sample preparation with mass spectrometry analysis in the selected reaction monitoring (SRM) mode. Absolute quantification of SEA was performed using the PSAQ™ method (Protein Standard Absolute Quantification), which uses a full-length isotope-labeled SEA as internal standard. The lower limit of detection (LLOD) and lower limit of quantification (LLOQ) were estimated at 352 pg/mL and 1057 pg/mL, respectively. SEA recovery after immunocapture was determined to be 7.8 ± 1.4%. Therefore, we assumed that less than 1 femtomole of each SEA proteotypic peptide was injected on the liquid chromatography column before SRM analysis. From a 6-point titration experiment, quantification accuracy was determined to be 77% and precision at LLOQ was lower than 5%. With this sensitive PSAQ-SRM assay, we expect to contribute to decipher the pathophysiological role of SEA in severe sepsis. This article is part of a Special Issue entitled: Proteomics: The clinical link.