Early regulation of brain aromatase (cyp19a1b) by estrogen receptors during zebrafish development.

Early expression of estrogen receptors (esr) and their role in regulating early expression of cyp19a1b encoding brain aromatase were examined in the brain of zebrafish. Using in toto hybridization and quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), a significant increase in the expression of esr1, esr2a, and esr2b was observed between 24 and 48 hours postfertilization (hpf). In toto hybridization demonstrated that esr2a and esr2b, but not esr1, are found in the hypothalamus. Using real-time RT-PCR, an increase in cyp19a1b mRNAs occurs between 24 and 48 hpf, indicating that expression of cyp19a1b is temporally correlated with that of esr. This increase is blocked by the pure anti-estrogen ICI182,780. Furthermore, E2 treatment of cyp19a1b-GFP (green fluorescent protein) transgenic embryos results in appearance of GFP expression in the brain as early as 25 hpf. These results indicate that basal expression of cyp19a1b expression in the brain of developing zebrafish most likely relies upon expression of esr that are fully functional before 25 hpf.

An operational approach to high resolution agro-ecological zoning in West-Africa.

The objective of this work is to develop a simple methodology for high resolution crop suitability analysis under current and future climate, easily applicable and useful in Least Developed Countries. The approach addresses both regional planning in the context of climate change projections and pre-emptive short-term rural extension interventions based on same-year agricultural season forecasts, while implemented with off-the-shelf resources. The developed tools are applied operationally in a case-study developed in three regions of Guinea-Bissau and the obtained results, as well as the advantages and limitations of methods applied, are discussed. In this paper we show how a simple approach can easily generate information on climate vulnerability and how it can be operationally used in rural extension services.

COUP-TFI (chicken ovalbumin upstream promoter-transcription factor I) regulates cell migration and axogenesis in differentiating P19 embryonal carcinoma cells.

The developmental expression patterns of the nuclear orphan receptors COUP-TFs (chicken ovalbumin upstream promoter-transcription factors) have been correlated to neurogenesis in several animal species. Nevertheless, the role of COUP-TFs in neurogenesis remains unknown. We have studied the functional involvement of COUP-TFI in retinoic acid (RA)-induced neuronal differentiation of P19 embryonal carcinoma cells through two complementary approaches: 1) deregulated expression of COUP-TFI, and 2) inactivation of endogenous COUP-TFs by means of a dominant-negative COUP-TFI mutant. Low levels of wild-type (wt)COUP-TFI transgene expression did not inhibit neural cell fate and primarily enhanced neuron outgrowth from RA-treated P19 aggregates. In contrast, high COUP-TFI expression impeded the neuronal differentiation of P19 cells induced with RA, resulting in cell cultures lacking neurons. This morphological effect was correlated to an elevated level of E-cadherin mRNA. The dominant-negative COUP-TFI mutant induced cell packing after RA treatment and inhibited neurite extension and neuron outgrowth from aggregates. A RGD peptide interference assay indicated that endogenous COUP-TFs could favor migration of neurons through an integrin-dependent mechanism. Accordingly, vitronectin mRNA levels were shown to be up-regulated by COUP-TFI by RT-PCR analysis, and COUP-TFI stimulated the mouse vitronectin promoter activity in transient transfection assays. Taken together, these data indicate that COUP-TFI is not simply a global repressor of retinoid functions, but shows a high selectivity for regulating genes involved in cellular adhesion and migration processes that are particularly important for neuronal differentiation.

Analysis of the estrogen regulation of the zebrafish estrogen receptor (ER) reveals distinct effects of ERalpha, ERbeta1 and ERbeta2.

We have previously cloned and characterized three estrogen receptors (ER) in the zebrafish (zfERalpha, zfERbeta1 and zfERbeta2). We have also shown that they are functional in vitro and exhibit a distinct expression pattern, although partially overlapping, in the brain of zebrafish. In this paper, we have shown that the hepatic expression of these zfER genes responds differently to estradiol (E2). In fact, a 48-h direct exposure of zebrafish to E2 resulted in a strong stimulation of zfERalpha gene expression while zfERbeta1 gene expression was markedly reduced and zfERbeta2 remained virtually unchanged. To establish the potential implication of each zfER in the E2 upregulation of the zfERalpha gene, the promoter region of this gene was isolated and characterized. Transfection experiments with promoter-luciferase reporter constructs together with different zfER expression vectors were carried out in different cell contexts. The data showed that in vivo E2 upregulation of the zfERalpha gene requires ERalpha itself and a conserved transcription unit sequence including at least an imperfect estrogen-responsive element (ERE) and an AP-1/ERE half site at the proximal transcription initiation site. Interestingly, although in the presence of E2 zfERalpha was the most potent at inducing the expression of its own gene, the effect of E2 mediated by zfERbeta2 represented 50% of the zfERalpha activity. In contrast, zfERbeta1 was unable to upregulate the zfERalpha gene whereas this receptor form was able to tightly bind E2 and activate a reporter plasmid containing a consensus ERE. Altogether, these results indicated that the two ERbeta forms recently characterized in teleost fish could have partially distinct and not redundant functions.

Accurate d-spacings from zero-order Laue zone patterns.

Experimental d-spacing values are criteria towards the identification of crystallites by electron diffraction. Conclusive identifications often rely on accurate d-spacings. It is shown here that accurate orthogonal components (in mm) for the primitive unit vectors of a zero-level diffraction pattern can be obtained through least-squares processing of (x,y) coordinates for all spots on the film. Valid vectors from the origin spot to any spot in the plane of the film are integer linear combinations of the two selected unit vectors. Accurate lengths and standard deviations for such vectors therefore can be calculated from the least-squares results. Corresponding d-spacings can then be calculated from the vector lengths on the film and the camera constant. In order to obtain d-spacing values that are not only precise, but also accurate, an accurate value of the camera constant should be used. This requires calibration of the experimental setup with reference materials in the same experimental conditions, with careful control of the sample height. For the same quality of measurements, the improvement in the accuracy of the d-spacings obtained with the proposed method is approximately proportional to the square root of the number of measurements taken. Practically, typical improvement in accuracy is about threefold, and accuracies of a fraction of a percent in d-spacings are achievable in this way. The above approach has been programmed as an option in the NRCBED program.

Ab-initio primitive cell parameters from single convergent-beam electron diffraction patterns: a converse route to the identification of microcrystals with electrons.

A new method for the ab initio derivation of Buerger-reduced primitive cell parameters from coordinate measurements of spots on single convergent-beam electron diffraction (CBED) patterns is described, which does not involve trial-and-error. The pattern can be taken along any zone axis, and misorientations of the crystallite by as much as a few degrees are taken into account without loss of accuracy. This derivation of cell parameters by least-squares analysis of the measurements has been automated in a program called NRCBED. Present accuracy is about 1% on lengths and 2 degrees on angles, but could be significantly improved by modelling projector lens aberrations, or by using a microscope without a projector lens. With present technology, it is possible to obtain a CBED pattern and a semi-quantitative energy-dispersive X-ray (EDX) analysis simultaneously from a single microcrystal a few hundred Angströms across. It becomes therefore possible to identify the material of the crystal on a single CBED pattern: a cell parameter database for known compounds is searched with the primitive cell parameters obtained in the above way, and with a mask describing the EDX results qualitatively. Feasibility is demonstrated on a crystallite of CeO2 500 Angströms across. With this new approach, trial-and-error should disappear from the solution of other long-standing problems: interpretation of X-ray powder patterns for new compounds in the presence of impurity lines, or in the case of multiple phases should become straight-forward.

Understorey fire frequency and the fate of burned forests in southern Amazonia.

Recent drought events underscore the vulnerability of Amazon forests to understorey fires. The long-term impact of fires on biodiversity and forest carbon stocks depends on the frequency of fire damages and deforestation rates of burned forests. Here, we characterized the spatial and temporal dynamics of understorey fires (1999-2010) and deforestation (2001-2010) in southern Amazonia using new satellite-based estimates of annual fire activity (greater than 50 ha) and deforestation (greater than 10 ha). Understorey forest fires burned more than 85 500 km(2) between 1999 and 2010 (2.8% of all forests). Forests that burned more than once accounted for 16 per cent of all understorey fires. Repeated fire activity was concentrated in Mato Grosso and eastern Pará, whereas single fires were widespread across the arc of deforestation. Routine fire activity in Mato Grosso coincided with annual periods of low night-time relative humidity, suggesting a strong climate control on both single and repeated fires. Understorey fires occurred in regions with active deforestation, yet the interannual variability of fire and deforestation were uncorrelated, and only 2.6 per cent of forests that burned between 1999 and 2008 were deforested for agricultural use by 2010. Evidence from the past decade suggests that future projections of frontier landscapes in Amazonia should separately consider economic drivers to project future deforestation and climate to project fire risk.

Endocrine disrupters: a review of some sources, effects, and mechanisms of actions on behaviour and neuroendocrine systems.

Some environmental contaminants interact with hormones and may exert adverse consequences as a result of their actions as endocrine disrupting chemicals (EDCs). Exposure in people is typically a result of contamination of the food chain, inhalation of contaminated house dust or occupational exposure. EDCs include pesticides and herbicides (such as dichlorodiphenyl trichloroethane or its metabolites), methoxychlor, biocides, heat stabilisers and chemical catalysts (such as tributyltin), plastic contaminants (e.g. bisphenol A), pharmaceuticals (i.e. diethylstilbestrol; 17α-ethinylestradiol) or dietary components (such as phytoestrogens). The goal of this review is to address the sources, effects and actions of EDCs, with an emphasis on topics discussed at the International Congress on Steroids and the Nervous System. EDCs may alter reproductively-relevant or nonreproductive, sexually-dimorphic behaviours. In addition, EDCs may have significant effects on neurodevelopmental processes, influencing the morphology of sexually-dimorphic cerebral circuits. Exposure to EDCs is more dangerous if it occurs during specific 'critical periods' of life, such as intrauterine, perinatal, juvenile or puberty periods, when organisms are more sensitive to hormonal disruption, compared to other periods. However, exposure to EDCs in adulthood can also alter physiology. Several EDCs are xenoestrogens, which can alter serum lipid concentrations or metabolism enzymes that are necessary for converting cholesterol to steroid hormones. This can ultimately alter the production of oestradiol and/or other steroids. Finally, many EDCs may have actions via (or independent of) classic actions at cognate steroid receptors. EDCs may have effects through numerous other substrates, such as the aryl hydrocarbon receptor, the peroxisome proliferator-activated receptor and the retinoid X receptor, signal transduction pathways, calcium influx and/or neurotransmitter receptors. Thus, EDCs, from varied sources, may have organisational effects during development and/or activational effects in adulthood that influence sexually-dimorphic, reproductively-relevant processes or other functions, by mimicking, antagonising or altering steroidal actions.

Primitive unit cell volumes obtained from unindexed convergent-beam electron diffraction patterns.

Provided that multiple reflection is present, a common occurrence with electron diffraction on microcrystallites, a single unindexed convergent-beam electron diffraction (CBED) pattern taken with the beam axis parallel to any direct lattice row allows the volume of the primitive cell to be calculated. The film measurements required are the diameters of the successive high-order Laue zones (HOLZ), the lengths of any two coprime vectors in the zero-order Laue zone (ZOLZ), and the angle between them. The primitive cell volume is an objective criterion allowing in a simplification in the identification of a phase under study by rapidly eliminating other possible phases. The computer program CELVOL for the calculation of the primitive cell volume from film measurements, or from literature cells, is available from the authors.

Molecular structure of opiate alkaloids. IV. Structure of two thioniamorphinans.

(I) S(equatorial)-Allyl-3-hydroxy-17-thioniamorphinan perchlorate, C19H25OS+.C1O4-, Mr = 400.91, monoclinic, P2(1)/n, a = 9.4171 (8), b = 10.7425 (10), c = 19.096 (2) A, beta = 95.666 (7) degrees, V = 1922.4 (3) A3, Z = 4, Dx = 1.385 Mg m-3, lambda (Mo K alpha) = 0.70930 A, mu = 0.33 mm-1, F(000) = 847.92, room temperature, final R = 0.052 for 1742 observed reflections. (II) S(axial)-Allyl-3-hydroxy-17-thioniamorphinan perchlorate, C19H25OS+.C1O4-, Mr = 400.91, monoclinic, P2(1), a = 8.4554 (10), b = 11.658 (3), c = 9.5831 (21) A, beta = 95.620 (10) degrees, V = 940.1 (3) A 3, Z = 2, Dx = 1.416 Mg m-3, lambda (Mo K alpha) = 0.70930 A, mu = 0.33 mm-1, F(000) = 423.96, room temperature, final R = 0.054 for 1015 observed reflections. The molecular structures of (I) and (II) are differentiated only by the orientation of the S-allyl substituent: the S-allyl group is equatorial in (I) and axial in (II). It has been shown that the activities as potent and selective blockers of kappa opioid receptors (kappa 2 subtype) are to be attributed to the alpha-thiamorphinan isomer (I). The inactive compound (II) is the beta-isomer. The fused ring systems are almost identical in the two molecules.