Inhibition by prolactin of gonadotrophin-induced cyclic AMP accumulation in the preovulatory ovary

We investigated whether prolactin acts at the ovarian level by interfering with the accumulation of gonadotrophin-induced ovarian cyclic AMP. Mouse ovaries were incubated with hCG and varying doses of prolactin. At the end of the incubation, the cyclic AMP which accumulated in the tissue + medium was measured. In ovaries devoid of corpora lutea, a significant inverse correlation (r = -0.93, P less than 0.05) was obtained between the doses of prolactin (0.1-25.6 micrograms ovine prolactin) and hCG-induced accumulation of ovarian cyclic AMP. In the presence of the phosphodiesterase inhibitor IBMX, however, the same doses of prolactin failed to exhibit any restricting influence on the accumulation of cyclic AMP. In luteinized ovaries, the same doses of prolactin in the absence of IBMX did not inhibit the hCG-induced cyclic AMP accumulation.     

Environmental influences on the qualitative and quantitative composition of phytoplankton and zooplankton in North African coastal lagoons

Within the framework of the international research project MELMARINA, seasonal dynamics of plankton communities in three North African coastal lagoons (Merja Zerga, Ghar El Melh, and Lake Manzala) were investigated. The sampling period extended from July 2003 to September 2004 with the aim of evaluating hydrological and other influences on the structure, composition and space-time development of these communities in each lagoon. Phytoplankton in Merja Zerga showed a quasi-permanent predominance of marine diatoms in the open sea station and in the marine inlet channel. Dinoflagellates were abundant in summer and early autumn in the marine inlet and extended into the central lagoon station. In Ghar El Melh, marine species (especially diatoms and dinoflagellates) dominated despite occasional winter inflows of freshwater. In Lake Manzala, freshwater species generally predominated and the planktonic communities were comparatively very diverse. Chlorophyceae contributed 39% of the total species recorded and diatoms and cyanophyceans were also common; the Dinophyceae, Euglenophyceae, Chrysophyceae and Cryptophyceae less so. Zooplankton communities in both Ghar El Melh and Merja Zerga were dominated by marine copepods. Rotifera, Copepoda, Ostracoda, and Cladocera were recorded in both lagoons as were meroplanktonic larvae of Polychaeta, Cirripedia, Mysidacea and Gastropoda and free living nematodes. Ghar El Melh was the more productive of these two lagoons with spring and early summer being the productive seasons. Zooplankton communities in Lake Manzala were generally dominated by rotifers and highest zooplankton abundances occurred in April (2003). Sampling stations near the marine inlets showed the highest diversity and the zooplankton communities showed considerable spatial variation within this large lagoon. The three lagoons represent very different water bodies contrasted strongly in terms of tidal effects and freshwater availability. Yet, there are some similarities in ecosystem structure. Space-time development of the plankton communities was similar especially in Merja Zerga and Ghar El Melh. Species abundances and specific diversities indicated that seasonal changes in salinity and nutrient concentrations were the main influential factors. Lake Manzala was the most productive lagoon and all the three sites supported toxic algal species. Relatively low plankton biomass in Merja Zerga and Ghar El Melh probably resulted from a combination of factors including highly episodic nutrient inputs, light suppression (by turbidity) and nutrient competition with benthic algae. Water quality variables were largely driven by the hydrological regime specific to each lagoon. Nutrient enrichment and, particularly for Lake Manzala, sea level rise threaten the sustainability of the planktonic ecosystems in all three lagoons. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Guest editors: J. R. Thompson & R. J. Flower Hydro-ecological Monitoring and Modelling of North African Coastal Lagoons     

Anti-thyroid cancer properties of a novel isoflavone derivative, 7-(O)-carboxymethyl daidzein conjugated to N-t-Boc-hexylenediamine in vitro and in vivo

The incidence of thyroid cancer is up to 3 folds higher in women than in men, suggesting that estrogenic effects may be involved in the pathogenesis of this malignancy. Here, we explore whether or not human thyroid cancer cell growth can be curbed by a novel isoflavone derivative generated in our laboratory, the N-t-Boc-hexylenediamine derivative of 7-(O)-carboxymethyl daidzein (cD-tboc). With the exception of the follicular cancer cell line WRO, estrogen receptor (ER)α mRNA was only marginally expressed in cell lines derived from papillary (NPA), follicular (MRO), anaplastic thyroid carcinoma (ARO) such that the expression of estrogen receptor (ER) βmRNA was more abundant than that of ERα mRNA in these cell types. Estradiol-17β (E2; 0.03-300nmol/l) per se increased proliferation in all four cell-types. The ERβ-specific agonist DPN increased [(3)H]-thymidine incorporation in all four thyroid cancer cell lines, whereas the ERα-specific agonist PPT increased growth only in NPA and WRO. By contrast, cD-tboc, derived from the weak estrogen daidzein, did not cause cell growth and dose-dependently diminished cell growth in all four cell lines via apoptosis and not necrosis, as detected by the release of histone-DNA fragments. The cytotoxic growth inhibitory effect of cD-tboc in these cells was modulated by E2 and the general caspase inhibitor Z-VAD-FMK, and the magnitude of this salvage was cell type-and dose-dependent. When nude mice carrying ARO thyroid xenografts were treated with cD-tboc, tumor volume decreased significantly, and no apparent toxicity was observed. These results suggest that cD-tboc may be a promising agent for therapy of thyroid carcinoma either alone or in combination with existing cytotoxic drugs.     

Ecological and genetic differentiation of Barbus callensis populations in Tunisia

Enzyme electrophoresis on horizontal starch gel was carried out on 356 barbel individuals. The sampling comprised 278 individuals of the species Barbus callensis from 10 rivers in Tunisia. The other individuals belonged to reference species (outgroups) from France and Morocco. An ecological study was also carried out on Tunisian rivers. The results show a clear differentiation of the two samples from northwestern Tunisia, which was only partly correlated with ecological characteristics of the rivers they inhabit. There is no genetic cline, but rather a discontinuity between populations in the northwestern-most watershed and all the other Tunisian populations. This differentiation probably has a paleohistoric origin not only related to adaptation to ecological conditions but also to difficulties in colonizing the watersheds. The results do not indicate clearly a colonization direction for the genus Barbus in North Africa. Analysis of the Algerian populations would appear to be indispensable. Lastly, in contrast with the usual taxonomy, Morocco and Tunisia are populated by two closely related species, but B. callensis should remain the name of the Tunisian species, which was the first to be described in the small El Kebir basin, a river that flows from Tunisia to Algeria.     

Sediment distribution and accumulation in lagoons of the Southern Mediterranean Region (the MELMARINA Project) with special reference to environmental change and aquatic ecosystems

Surface sediments and sediment cores were collected from coastal lagoons and lakes located in the Southern Mediterranean Region (SMR) as part of the MELMARINA Project which involved integrated eco-hydrological monitoring and modelling. This study uses surface sediments and sediment cores to infer spatial characteristics and temporal changes at the MELMARINA primary sites, Merja Zerga in Morocco, Ghar El Melh in Tunisia and Lake Manzala in Egypt. In addition, surface sediment sampling was undertaken at Egyptian Lake Bardawil and sediment cores were collected from the Lagune de Nador (Morocco). Sediment distribution patterns are investigated using GIS with georeferenced sample locations to facilitate display and resurvey. Major variations in sedimentary organic matter and, particularly, carbonate content, occur within and between sites. Local landscapes combined with hydrological and biogeochemical processes influence the distributions of sediment bulk components (carbonates, organic material and clastic matter) and molluscan shells and shell debris are an important source of sedimentary carbonate at all three primary sites. Sediment cores were dated using natural (²¹⁰Pb) and artificial (¹³⁷Cs) radionuclides, and sediment accumulation rate changes indicate that sources of sediment supply varied markedly through the twentieth century but have generally diminished after the mid-1960s. Sedimentary siliceous microfossils (diatoms) were generally poorly preserved, but mollusc shell remains were well represented. Sediment chronologies and sediment bulk composition allow discussion of some recent changes in bulk, minerogenic and biogenic sediment accumulation patterns in the SMR lagoons. Sediment accumulation rates also varied between sites and multiple cores from Lake Manzala indicated that rates showed considerable spatial variability. Low-level sediment contamination by fossil fuel combustion particulates and trace metals was demonstrated for Ghar El Melh and Lagune de Nador where Pb and Zn accumulation rates were highest in twentieth century sediment. It is emphasized that sediment quality and quantity have strong influences on lagoon ecosystem function and sedimentation is relevant to hydromorphology and to concepts of ecological quality. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Guest editors: J. R. Thompson & R. J. Flower Hydro-ecological Monitoring and Modelling of North African Coastal Lagoons     

Engineering a potential antagonist of human thyrotropin and thyroid-stimulating antibody

Thyrotropin (TSH) and the gonadotropins (FSH, LH, hCG) are a family of heterodimeric glycoprotein hormones composed of two noncovalently linked subunits, alpha and beta. We have recently converted the hTSH heterodimer to a biologically active single chain (hTSHbeta.CTPalpha) by fusing the common alpha-subunit to the C-terminal end of the hTSH beta-subunit in the presence of a approximately 30-amino acid peptide from hCGbeta (CTP) as a linker. The hTSHbeta.CTPalpha single chain was used to investigate the role of the N-linked oligosaccharides of alpha- and beta-subunits in the secretion and function of hTSH. Using overlapping PCR mutagenesis, two deglycosylated variants were prepared: one lacking both oligosaccharide chains on the alpha-subunit (hTSHbeta.CTPalpha(1+2)) and the other lacking the oligosaccharide chain on the beta-subunit (hTSHbeta.CTPalpha(deg)). The single chain variants were expressed in CHO cells and were secreted into the medium. hTSH variants lacking the oligosaccharide chains were less potent than hTSHbeta.CTPalpha wild-type with respect to cAMP formation and thyroid hormone secretion in cultured human thyroid follicles. Both deglycosylated variants competed with hTSH in a dose-dependent manner. The hTSHbeta.CTPalpha(1+2) variant blocked cAMP formation and thyroid hormone secretion stimulated by hTSH as well as by the antibody, thyroid-stimulating immunoglobulins, responsible for the most common cause of hyperthyroidism, Graves disease. Thus, this variant behaves as a potential antagonist, offering a novel therapeutic strategy in the treatment of thyrotoxicosis caused by Graves' disease and TSH-secreting pituitary adenoma.     

Effects of prolactin on the morphology of cultured rat granulosa cells

Morphological changes were correlated with biochemical data induced by prolactin (PRL) in cultured rat granulosa cells from large preovulatory follicles. Biochemical results indicated that PRL exerted a significant dose-dependent inhibition in gonadotrophin-induced secretion of progesterone and 17 beta-oestradiol. PRL alone failed to affect basal steroidogenic secretion. In parallel morphological experiments, using phase-contrast microscopy, untreated and 100 ng/ml PRL-treated cells appeared as a monolayer of flattened, fibroblast-like cells. Upon exposure to 0.4 IU/ml human chorionic gonadotrophin (hCG), aggregates of rounded, epithelioid-shaped cells were formed. The addition of PRL to hCG in the same doses minimized the changes induced by hCG. Similarly, electron microscopy of untreated and PRL-treated cultures revealed flat cells devoid of microvilli, with evenly dispersed microfilaments. The addition of hCG caused rounding of the cells and was accompanied by the appearance of microvilli and by pronounced steroid-producing organelles. Bundles of microfilaments were noted at the cell periphery. PRL added to hCG caused a reduction of the hCG effects, and the cell morphology was intermediate to that seen in untreated and hCG-treated cultures. The finding that PRL can prevent or minimize morphological changes caused by hCG in rat cultured granulosa cells correlates with the biochemical changes induced by PRL, and supports the concept that PRL is a modulator of gonadotrophic action in the ovary.     

Size‐dependent effects of daily thermal fluctuations on the growth and size heterogeneity of Nile tilapia Oreochromis niloticus

The growth of Nile tilapia Oreochromis niloticus (0·02–20·00 g) was measured when fed to excess during the hours of light, following their exposure to five thermal regimes fluctuating around the thermal optimum for growth (T_opt = 30° C) over the diel cycle of day (light, L) and night (dark, N), i.e. 27° C(L):33° C(N), 28·5° C(L):31·5° C(N), 30° C(L):30° C(N), 31·5° C(L):28·5° C(N) and 33° C(L):27° C(N) (two replicates per treatment, six weeks' rearing, growth measurements at weekly intervals). A model constructed with a stepwise multiple‐regression analysis accounted for 87·4% of the variation of the specific growth rate (G, % M day^?1) from the variations of wet mass (M), the extent of the thermal fluctuation (F_T) and their interactions, i.e. log₁₀G = 1·7686 ? 0·2136 log₁₀M + 0·0806 [log ₁₀M× log ₁₀ (1 + F_T)] ? 0·0394 [log₁₀M× log ₁₀ (1 + F_T)]². Based on this model, the thermal fluctuation that produces the fastest growth ( ,°C) decreases in a curvilinear way, from 5·1° C at 20 mg to c. 0·7° C at 20 g. Thermal regimes that produce the slowest growth also produce the highest size heterogeneity. Functional hypotheses behind the size‐dependent effects of thermal fluctuations are discussed, together with their implications in natural habitats and aquaculture systems with in different contexts of food availability.     

Impaired light chain allelic exclusion and lack of positive selection in immature B cells expressing incompetent receptor deficient of CD19

Positive signaling is now thought to be important for B cell maturation, although the nature of such signals has not yet been defined. We are studying the regulatory role of B cell Ag receptor (BCR) signaling in mediating positive selection of immature B cells. To do so, we use Ig transgenic mice (3-83Tg) that are deficient in CD19, thus generating a monoclonal immature B cell population expressing signaling-incompetent BCR. Immature 3-83Tg CD19(-/-) B cells undergo developmental arrest in the bone marrow, allowing maturation only to cells that effectively compensate for the compromised receptor by elevated levels of BCR. We find that developmentally arrested 3-83Tg CD19(-/-) B cells fail to impose L chain allelic exclusion and undergo intensive V(D)J recombination to edit their BCR. Furthermore, immature 3-83Tg CD19(-/-) B cells, which were grown in vitro, failed to undergo positive selection and to survive when adoptively transferred into normal recipients. However, elevation of BCR expression levels, obtained by transgene homozygosity, effectively compensated for the compromised BCR and completely restored BCR-mediated Ca(2+) influx, allelic exclusion, and positive selection. Our results suggest that the BCR signaling threshold mediates positive selection of developing B cells, and that a receptor-editing mechanism has an important role in rescuing cells that fail positive selection because of incompetent receptors.