Purification, crystallization and preliminary X-ray analysis of a fusion of the LIM domains of LMO2 and the LID domain of Ldb1

LMO2 (LIM domain only 2), also known as rhombotin-2, is a transcriptional regulator that is essential for normal haematopoietic development. In malignant haematopoiesis, its ectopic expression in T cells is involved in the pathogenesis of leukaemia. LMO2 contains four zinc-finger domains and binds to the ubiquitous nuclear adaptor protein Ldb1 via the LIM-interaction domain (LID). Together, they act as scaffolding proteins and bridge important haematopoietic transcription factors such as SCL/Tal1, E2A and GATA-1. Solving the structure of the LMO2:Ldb1-LID complex would therefore be a first step towards understanding how haematopoietic specific protein complexes form and would also provide an attractive target for drug development in anticancer therapy, especially for T-cell leukaemia. Here, the expression, purification, crystallization and data collection of a fusion protein consisting of the two LIM domains of LMO2 linked to the LID domain of Ldb1 via a flexible linker is reported. The crystals belonged to space group C2, with unit-cell parameters a = 179.9, b = 51.5, c = 114.7 Å, β = 90.1°, and contained five molecules in the asymmetric unit. Multiple-wavelength anomalous dispersion (MAD) data have been collected at the zinc X-ray absorption edge to a resolution of 2.8 Å and the data were used to solve the structure of the LMO2:Ldb1-LID complex. Refinement and analysis of the electron-density map is in progress

The histone H3K4 demethylase JARID1A directly interacts with haematopoietic transcription factor GATA1 in erythroid cells through its second PHD domain

Chromatin remodelling and transcription factors play important roles in lineage commitment and development through control of gene expression. Activation of selected lineage-specific genes and repression of alternative lineage-affiliated genes results in tightly regulated cell differentiation transcriptional programmes. However, the complex functional and physical interplay between transcription factors and chromatin modifying enzymes remains elusive. Recent evidence has implicated histone demethylases in normal haematopoietic differentiation as well as in malignant haematopoiesis. Here we report an interaction between H3K4 demethylase JARID1A and the haematopoietic-specific master transcription proteins SCL and GATA1 in red blood cells. Specifically, we observe a direct physical contact between GATA1 and the second PHD domain of JARID1A. This interaction has potential implications for normal and malignant haematopoiesis

Metamorphic evolution and U-Pb zircon SHRIMP geochronology of the Belizario ultramafic amphibolite, Encantadas Complex, southernmost Brazil.

The integrated investigation of metamorphism and zircon U-Pb SHRIMP geochronology of the Belizario ultramafic amphibolite from southernmost Brazil leads to a better understanding of the processes involved in the generation of the Encantadas Complex. Magmatic evidence of the magnesian basalt or pyroxenite protolith is only preserved in cores of zircon crystals, which are dated at 2257 ± 12 Ma. Amphibolite facies metamorphism M1 formed voluminous hornblende in the investigated rock possibly at 1989 ± 21 Ma. This ultramafic rock was re-metamorphosed at 702 ± 21 Ma during a greenschist facies event M2; the assemblage actinolite + oligoclase + microcline + epidote + titanite + monazite formed by alteration of hornblende. The metamorphic events are probably related to the Encantadas Orogeny (2257 ± 12 Ma) and Camboriu Orogeny (~ 1989 Ma) of the Trans-Amazonian Cycle, followed by an orogenic event (702 ± 21 Ma) of the Brasiliano Cycle. The intervening cratonic period (2000-700 Ma) corresponds to the existence of the Supercontinent Atlantica, known regionally as the Rio de la Plata Craton

THE OPTICAL PROPERTIES OF Pr3+ EMBEDDED IN THE RARE EARTH BOROGERMANATE MATRICES: REBGeO5

The luminescent properties of the trivalent praseodymium ion in the trigonal borogermanate matrice PrBGeO5 have been analysed. The energy level schemes are deduced from the absorption and emission spectra and reproduced with 14 crystal field parameters (cfps) according to the local point symmetry occupied by the rare earth element in the matrix.The luminescent properties of the trivalent praseodymium ion in the trigonal borogermanate matrice PrBGeO5 have been analysed. The energy level schemes are deduced from the absorption and emission spectra and reproduced with 14 crystal field parameters (cfps) according to the local point symmetry occupied by the rare earth element in the matrix

Activation of the hsp70 promoter by environmental inorganic and organic chemicals: relationships with cytotoxicity and lipophilicity.

International audienceStress proteins (heat shock proteins, HSPs) have been proposed as general markers of cellular aggression and their use for environmental monitoring is often suggested. The aim of this work was to study the potency of various environmentally relevant organic and inorganic chemicals to induce the expression of the HSP70 marker. For this purpose, we used an established HeLa cell line containing the chloramphenicol acetyl transferase (CAT) gene under the control of the hsp70 promoter. The screening of three metallic and 15 organic chemicals revealed differences in their capacities to induce the hsp70 promoter. The three metals tested (cadmium, zinc and mercury) were able to induce a stress response. Some organochlorine compounds (chlorophenol derivatives, tetrachlorohydroquinone, 3, 4-dichloroaniline, ethyl parathion and 1-chloro-2,4-dinitrobenzene) induced a response, whereas other common halogenated pesticides or aromatic hydrocarbons (e.g. benzo(a)pyrene, 2, 4-dichlorophenoxyacetic acid, endosulfan, diuron, 4-nonylphenol) did not. The potency to induce hsp70 was significantly correlated to the octanol-water partition coefficient (log K(ow)) of the inducing chemicals, except for 1-chloro-2,4-dinitrobenzene and ethyl parathion. Cytotoxicity assays run in parallel to the induction measurements revealed that the three metals were effective at non cytotoxic doses whereas all organic compounds, except tetrachlorohydroquinone and 1-chloro-2,4-dinitrobenzene, induced the promoter at cytotoxic doses. These results suggest that hsp70 is induced by different mechanisms of toxicity. We propose that this model can be used in mechanistic studies for the detection of toxic effects of certain pollutants

Exemples de résultats de la recherche obtenus sur les populations sauvages de salmonides de Bretagne et de Basse-Normandie et applicables à leur gestion

Ces dernières années, un ensemble de méthodes et techniques d'étude a été mis au point ou amélioré à partir de recherches réalisées sur les populations sauvages de salmonides. Dans un premier temps, ces nouvelles techniques peuvent être considérées comme des outils disponibles pour les gestionnaires; ce sont : la mise en place et l'homogénéisation de critères scalimétriques pour la détermination de l'âge des salmonidés migrateurs; la détermination du sexe par sérodiagnostic et à partir de critères morphométriques; l'estimation de la production en smolts de saumon à partir de l'habitat et des caractéristiques des peuplements automnaux. Dans un deuxième temps, l'application de certaines de ces méthodes dans le domaine écologique a permis d'obtenir des éléments essentiels pour la connaissance et/ou la gestion des populations sauvages de salmonides.Turing the fast few years some new techniques for the study of wild populations of salmonids have been developed. The following procedures are intended to provide.1) an improved method for determining age by scale-reading, particularly with migratory salmonids (Atlantic salmon and sea trout). A standard zone for removing scales, the presence of three different zones on each scale (river, run-out, sea) and some criteria for scale interpretation have all been defined. These criteria are mainly concerned with the structure of the scale during the sea-life stage and reproduction. On the other hand, a spawning mark has been determined for both sedentary and migratory salmonids.2) an means of sex determination. Two techniques have been proposed :- the first one is based on the detection by immunoagglutination of the serum vitellogenin of females in active gametogenesis. This method concerns females only; it does not allow a distinction to he made between non spermating males and immature salmonids. It is a very reliable technique which can be used in field conditions, but only by experienced researchers;- the second technique is based on the sexual dimorphism of the relative length of the upper jaw bone. It is very reliable (8 % to 12 % error) and can also be applied in field conditions, but only for large fish (fork length > 40-45 cm).3) a way of estimating Atlantic salmon smolt production. This technique is based on :- the estimation of a 0+ and 1+ salmon population in the whole river from data of density obtained in sectors representative of the different types of habitat ;- the presence of growth bimodality in a 0+ salmon population in autumn, given the following hypothesis : one yearold smolts belong to the upper mode ;- the estimation of mortality for the whole winter season.This technique has been used in the River Oir and compared with a smolt-trapping experiment conducted for two years. A comparison of the results shows that :- the one year-old smolts belong to the upper mode of bimodality;- when based on data on habitat characteristics, this technique can be used in field conditions to evaluate the smolts with sufficient accuracy.All three techniques are immediately available as useful methods for fisheries management. At a second stage, they can also serve in ecological research to advance knowledge on which to base further study and improved management of wild salmonid populations

Perspectives: Entrepreneurship Training Can Empower Students Being Left Behind

Entrepreneurial self-employment, however, would hold great promise for business-minded students, if they learn entrepreneurship in high school and can test out their innovative business plans on consumers in their own neighborhoods and beyond — especially Internet start-up ideas. The social and community networking success of MySpace opens a wide door for anyone to market a new idea or product to a myriad of potential customers instantly

Effects of human pharmaceuticals on cytotoxicity, EROD activity and ROS production in fish hepatocytes.

Pharmaceuticals are found in the aquatic environment but their potential effects on non-target species like fish remain unknown. This in vitro study is a first approach in the toxicity assessment of human drugs on fish. Nine pharmaceuticals were tested on two fish hepatocyte models: primary cultures of rainbow trout hepatocytes (PRTH) and PLHC-1 fish cell line. Cell viability, interaction with cytochrome P450 1A (CYP1A) enzyme and oxidative stress were assessed by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrasodium bromide tetrazolium (MTT), 7-ethoxyresorufin-o-deethylase (EROD) and dichlorofluorescein (DCFH-DA) assays, respectively. The tested drugs were clofibrate (CF), fenofibrate (FF), carbamazepine (CBZ), fluoxetine (FX), diclofenac (DiCF), propranolol (POH), sulfamethoxazole (SFX), amoxicillin (AMX) and gadolinium chloride (GdCl(3)). All substances were cytotoxic, except AMX at concentration up to 500 microM. The calculated MTT EC(50) values ranged from 2 microM (CF) to 651 microM (CBZ) in PLHC-1, and from 53 microM (FF) to 962 microM (GdCl(3)) in PRTH. CF, FF, and FX were the most cytotoxic drugs and induced oxidative stress before being cytotoxic. Compared to hepatocytes from human and dog, fish hepatocytes seemed to be more susceptible to the peroxisome proliferators (PPs) CF and FF. In PLHC-1 cells none of the tested drugs induced the EROD activity whereas POH appeared as a weak EROD inducer in PRTH. Moreover, in PRTH, SFX, DiCF, CBZ and to a lesser extend, FF and CF inhibited the basal EROD activity at clearly sublethal concentrations which may be of concern at the biological and chemical levels in a multipollution context