Endocrine disruption in the Scheldt estuary distribution, exposure and effects (ENDIS-RISKS). Final report

ENDIS-RISKS is a multidisciplinary, research project conducted by five institutes. This project aimed to assess the distribution, exposure and effects of endocrine disruptors in the Scheldt estuary, with specific attention to invertebrates. The Scheldt estuary is known to be one of the most polluted estuaries in the world. The industrial areas of Ghent and Antwerp are to a large extent responsible for this pollution. To achieve these goals detailed knowledge of the distribution and long-term effects of these substances is needed. This information is crucial for the development of future-oriented policy measures at the national and European level. The project can be divided into four different research phases. In Phase I the occurance and distribution of endocrine disrupting substances in the Scheldt estuary was studied. Water, sediment, suspended solids and biota were sampled 3 times a year for a period of 4 years (2002-2006). In all these matrices, 7 groups of chemicals were analysed: estrogens, pesticides, phthalates, organotins, polyaromatic components (PCBs, PBDEs), polyaromatic hydrocarbons (PAHs) and phenols. All the analyzed chemicals are on the OSPAR list of priority chemicals or are indicated as endocrine disruptors on this list. The different water samples were also tested using in vitro assays to assess their potential to bind to the (human) estrogen and androgen receptor. Phase II evaluated the exposure of biota occuring in the Scheldt estuary to endocrine disrupting substances. Based on the results of the chemical analysis, priority substances were selected. Phase III studied the effects of endocrine disrupting substances occurring in the Scheldt estuary on resident mysid shrimp populations (laboratory and field studies). Substances of concern were selected and tested in the laboratory to evaluate their effects on the estuarine mysid Neomysis integer. In the context of this project, three new assays using invertebrate-specific endpoints were developed to examine the effect of endocrine disrupting chemicals (EDCs) on molting, embryogenesis and vitellogenesis of N. integer. Finally, in Phase IV laboratory and field results were used to perform a preliminary environmental risk assessment of endocrine disruptors in the Scheldt estuary. Samples were collected along the salinity gradiënt of the Scheldt estuary with the RV Belgica. Water samples were taken with Teflon-coated Go-Flo bottles (10L), sediment samples with Van Veen Grab, biota with a hyperbentic sledge, and suspended particulate matter (SPM) was continuously sampled with an Alfa Laval flow-through centrifuge. For the chemical analysis, protocols were developed to analyse estrogens, organotriazine herbicides, organochlorine pesticides, phtalates, organotins, PAHs, PCBs, and PBDEs in the different matrices: i.e. water, sediment, SPM and biota.Experimental studies were performed to analyse growth, molting, embryogenesis and vitellogenesis of N. integer. These studies were needed to develop ecotoxicological assays to evaluate EDCs on these physiological processes. To study growth of N. integer, organisms were individually transferrred in exposure solutions and molts were collected to measure the growth after each molting. To study embryogenesis, embryos were taking out of the marsupium and placed in multiwell plates. Each day survival, developmental stages and hatching was analysed. To study vitellogenesis, vitellin was isolated from eggs with gelfitration and polyclonal antibodies were developed (in rabbits). With the isolated vitellin and the antibodies an enzyme-linked immunosorbent assay (ELISA) was developed. Vitellin was quatified in ovigerous females exposed to test compound in the laboratory and in females collected from the different sampling sites of the Scheldt estuary. In addition to vitellin levels, energy allocation and testosterone metabolism was examined in field collected mysids. Finally, results from population stu

Male-male marriage in Sinophone and Anglophone Harry Potter danmei and slash

The aim of this study is to compare Sinophone and Anglophone fan fiction consisting of female-oriented male-male romance: danmei and slash, respectively. To increase comparability, we analysed Harry Potter fan fiction in which the characters Harry and Draco are married. Male-male marriage was selected because our online Sinophone and Anglophone BL fandom surveys indicate this to be the most popular story element of the nine options we provided. We analysed five stories originally written in Chinese and five originally written in English which subsequently had been fan-translated into Chinese. Using Thematic Analysis (Braun & Clarke, 2006) we found some robust patterns. In contrast to the Anglophone fiction, the Sinophone tended to: stress the importance of family approval for the marriage; incorporate a wedding ceremony; employ clearly gendered roles between partners; utilise extended, as opposed to nuclear, families; and showed the couple to produce children, particularly boys. Hence, the stories mirror the relative social conservatism and social liberalism of their cultures of origin. However, in reading and writing such danmei young Chinese women are still pushing at the boundaries of the traditional family

Hadron Energy Reconstruction for the ATLAS Calorimetry in the Framework of the Non-parametrical Method

This paper discusses hadron energy reconstruction for the ATLAS barrel prototype combined calorimeter (consisting of a lead-liquid argon electromagnetic part and an iron-scintillator hadronic part) in the framework of the non-parametrical method. The non-parametrical method utilizes only the known $e/h$ ratios and the electron calibration constants and does not require the determination of any parameters by a minimization technique. Thus, this technique lends itself to an easy use in a first level trigger. The reconstructed mean values of the hadron energies are within $\pm 1$ of the true values and the fractional energy resolution is $[(58\pm3)/\sqrt{E}+(2.5\pm0.3)$. The value of the $e/h$ ratio obtained for the electromagnetic compartment of the combined calorimeter is $1.74\pm0.04$ and agrees with the prediction that $e/h > 1.7$ for this electromagnetic calorimeter. Results of a study of the longitudinal hadronic shower development are also presented. The data have been taken in the H8 beam line of the CERN SPS using pions of energies from 10 to 300 GeV.Comment: 33 pages, 13 figures, Will be published in NIM

Observation of the Early Structural Changes Leading to the Formation of Protein Superstructures.

Formation of superstructures in protein aggregation processes has been indicated as a general pathway for several proteins, possibly playing a role in human pathologies. There is a severe lack of knowledge on the origin of such species in terms of both mechanisms of formation and structural features. We use equine lysozyme as a model protein, and by combining spectroscopic techniques and microscopy with X-ray fiber diffraction and ab initio modeling of Small Angle X-ray Scattering data, we isolate the partially unfolded state from which one of these superstructures (i.e., particulate) originates. We reveal the low-resolution structure of the unfolded state and its mechanism of formation, highlighting the physicochemical features and the possible pathway of formation of the particulate structure. Our findings provide a novel detailed knowledge of such a general and alternative aggregation pathway for proteins, this being crucial for a basic and broader understanding of the aggregation phenomena.This is the author's accepted manuscript and will be under embargo until the 3rd of September 2015. The final version is published by ACS in The Journal of Physical Chemistry Letters here: http://pubs.acs.org/doi/abs/10.1021/jz501614e

Semi-automated Magnetic Bead-Based Antibody Selection from Phage Display Libraries

Phage display of combinatorial antibody libraries is a very efficient method for selecting recombinant antibodies against a wide range of molecules. It has been applied very successfully for the generation of therapeutic antibodies for more than a decade. To increase robustness and reproducibility of the selection procedure, we developed a semi-automated selection method for the generation of recombinant antibodies from phage display libraries. In this procedure, the selection targets are specifically immobilised to magnetic particles which can then by automatically handled by a magnetic particle processor. At present up to 96 samples can be handled simultaneously. Applying the processor allows standardisation of panning parameters such as washing conditions, incubation times, or to perform parallel selections on same targets under different buffer conditions. Additionally, the whole protocol has been streamlined to carry out bead loading, phage selection, phage amplification between selection rounds and magnetic particle ELISA for confirmation of binding activity in microtiter plate formats. Until now, this method has been successfully applied to select antibody fragments against different types of target, such as peptides, recombinant or homologous proteins, or chemical compounds

Performance of an endcap prototype of the Atlas accordion electromagnetic calorimeter

The design and construction of a lead-liquid argon endcap calorimeter prototype using an accordion geometry and conceived as a sector of the inner wheel of the endcap calorimeter of the future ATLAS experiment at the LHC is described. The performance obtained using electron beam data is presented. The main results are an energy resolution with a sampling term below $11\%/\sqrt{E(\rm GeV)}$ and a small local constant term, a good linearity of the response with the incident energy and a global constant term of 0.8\% over an extended area in the rapidity range of $2.2\eta 2.9$. These properties make the design suitable for the ATLAS electromagnetic endcap calorimeter