Characterization and Expression of Glutamate Dehydrogenase in Response to Acute Salinity Stress in the Chinese Mitten Crab, Eriocheir sinensis

Glutamate dehydrogenase (GDH) is a key enzyme for the synthesis and catabolism of glutamic acid, proline and alanine, which are important osmolytes in aquatic animals. However, the response of GDH gene expression to salinity alterations has not yet been determined in macro-crustacean species.GDH cDNA was isolated from Eriocheir sinensis. Then, GDH gene expression was analyzed in different tissues from normal crabs and the muscle of crabs following transfer from freshwater (control) directly to water with salinities of 16‰ and 30‰, respectively. Full-length GDH cDNA is 2,349 bp, consisting of a 76 bp 5'- untranslated region, a 1,695 bp open reading frame encoding 564 amino acids and a 578 bp 3'- untranslated region. E. sinensis GDH showed 64-90% identity with protein sequences of mammalian and crustacean species. Muscle was the dominant expression source among all tissues tested. Compared with the control, GDH expression significantly increased at 6 h in crabs transferred to 16‰ and 30‰ salinity, and GDH expression peaked at 48 h and 12 h, respectively, with levels approximately 7.9 and 8.5 fold higher than the control. The free amino acid (FAA) changes in muscle, under acute salinity stress (16‰ and 30‰ salinities), correlated with GDH expression levels. Total FAA content in the muscle, which was based on specific changes in arginine, proline, glycine, alanine, taurine, serine and glutamic acid, tended to increase in crabs following transfer to salt water. Among these, arginine, proline and alanine increased significantly during salinity acclimation and accounted for the highest proportion of total FAA.E. sinensis GDH is a conserved protein that serves important functions in controlling osmoregulation. We observed that higher GDH expression after ambient salinity increase led to higher FAA metabolism, especially the synthesis of glutamic acid, which increased the synthesis of proline and alanine to meet the demand of osmoregulation at hyperosmotic conditions

Pembauran Budaya di Desa Rantau Sakti Kecamatan Tambusai Utara Kabupaten Rokan Hulu

This study is to research how the intercourse of culture that existed in the Rantau Sakti village Tambusai Utara District and Rokan Hulu Regent intertribal nasions of Java, Medan Java, Batak, Minang, and Melayu. Theres a launching factors and hindrance factors too in culture intercourse. The data tekhnic collection with the koesioner, interview, and documentation. And the analisys of data is descriptive where the data presented in the form of a diagrams, percentages, pictures, and explanations. The research results show that The lives of the multicultural that could lead to the occurrence of intercourse of culture through assimilation, acculturation, and amalgamation. The intercourse of culture through three aspects of it can run well theres happened in Rantau Sakti village on seven principles like language, system of knowledge , a social organization , celebration life and technologi, means of subsistence, religion, and art . There is also intercourse of culture with amalgamation culture that leads to assimilation and acculturation. Its launching factor are the attitude of tolerance and open, in common religion, and the same occasion in the economic field. While the hindrance factor are differences of religion and stereotype (ethnic prejudice). Those two factors each happened to the process of changing culture with assimilation, acculturation, and amalgamation That leads to a positive thing.Keyword: The Culture Intercourse, Inter-ethnic

Behavioural and Physiological Responses of Gammarus pulex Exposed to Cadmium and Arsenate at Three Temperatures: Individual and Combined Effects

This study aimed at investigating both the individual and combined effects of cadmium (Cd) and arsenate (AsV) on the physiology and behaviour of the Crustacean Gammarus pulex at three temperatures (5, 10 and15°C). G. pulex was exposed during 96 h to (i) two [Cd] alone, (ii) two [AsV] alone, and (iii) four combinations of [Cd] and [AsV] to obtain a complete factorial plane. After exposure, survival, [AsV] or [Cd] in body tissues, behavioural (ventilatory and locomotor activities) and physiological responses (iono-regulation of [Na+] and [Cl−] in haemolymph) were examined. The interactive effects (antagonistic, additive or synergistic) of binary mixtures were evaluated for each tested temperature using a predictive model for the theoretically expected interactive effect of chemicals. In single metal exposure, both the internal metal concentration in body tissues and the mortality rate increased along metallic gradient concentration. Cd alone significantly impaired both [Na+] and [Cl−] while AsV alone had a weak impact only on [Cl−]. The behavioural responses of G. pulex declined with increasing metal concentration suggesting a reallocation of energy from behavioural responses to maintenance functions. The interaction between AsV and Cd was considered as ‘additive’ for all the tested binary mixtures and temperatures (except for the lowest combination at 10°C considered as “antagonistic”). In binary mixtures, the decrease in both ventilatory and locomotor activities and the decline in haemolymphatic [Cl−] were amplified when respectively compared to those observed with the same concentrations of AsV or Cd alone. However, the presence of AsV decreased the haemolymphatic [Na+] loss when G. pulex was exposed to the lowest Cd concentration. Finally, the observed physiological and behavioural effects (except ventilation) in G. pulex exposed to AsV and/or Cd were exacerbated under the highest temperature. The discussion encompasses both the toxicity mechanisms of these metals and their interaction with rising temperature

Expression of Calmodulin and Myosin Light Chain Kinase during Larval Settlement of the Barnacle Balanus amphitrite

Barnacles are one of the most common organisms in intertidal areas. Their life cycle includes seven free-swimming larval stages and sessile juvenile and adult stages. The transition from the swimming to the sessile stages, referred to as larval settlement, is crucial for their survivor success and subsequent population distribution. In this study, we focused on the involvement of calmodulin (CaM) and its binding proteins in the larval settlement of the barnacle, Balanus ( = Amphibalanus) amphitrite. The full length of CaM gene was cloned from stage II nauplii of B. amphitrite (referred to as Ba-CaM), encoding 149 amino acid residues that share a high similarity with published CaMs in other organisms. Quantitative real-time PCR showed that Ba-CaM was highly expressed in cyprids, the stage at which swimming larvae are competent to attach and undergo metamorphosis. In situ hybridization revealed that the expressed Ba-CaM gene was localized in compound eyes, posterior ganglion and cement glands, all of which may have essential functions during larval settlement. Larval settlement assays showed that both the CaM inhibitor compound 48/80 and the CaM-dependent myosin light chain kinase (MLCK) inhibitor ML-7 effectively blocked barnacle larval settlement, whereas Ca2+/CaM-dependent kinase II (CaMKII) inhibitors did not show any clear effects. The subsequent real-time PCR assay showed a higher expression level of Ba-MLCK gene in larval stages than in adults, suggesting an important role of Ba-MLCK gene in larval development and competency. Overall, the results suggest that CaM and CaM-dependent MLCK function during larval settlement of B. amphitrite

Polyamines, indolamines, and catecholamines in gills and haemolymph of the euryhaline crab, <i>Eriocheir sinensis</i>. Effects of high pressure and salinity

Tyrosine, octopamine, tryptophan, serotonin, putrescine, spermidine, spermine, norepinephrine, epinephrine, and dopamine were measured by means of high performance liquid chromatography (HPLC) in gills (anterior and posterior) and haemolymph extracts of the freshwater Chinese mitten crab, Eriocheir sinensis. These results may be considered as basic data to be used as control physiological doses, to which to refer for injection or for in vitro perfusion experiments. No significant difference has been detected between anterior respiratory and posterior salt-transporting gills. After 10 days acclimation to sea water, a significant increase of haemolymph norepinephrine and dopamine has been observed. Exposure to hydrostatic pressure (101 ATA for 5 hours) also results in an octopamine increase in the haemolymph. In addition, there is a tryptophan increase in both the anterior and the posterior gills. Results are discussed in terms of possible correlations of the concentrations of polyamines, indoleamines, and catecholamines with osmoregulation and behaviour

Combined estrogenic and anti-estrogenic properties of estetrol on breast cancer may provide a safe therapeutic window for the treatment of menopausal symptoms

Increased risk of breast cancer is a critical side effect associated with the use of a menopausal hormone therapy (MHT). Estetrol (E4) is a natural estrogen produced by the human fetal liver and is a promising compound for clinical use in MHT. However, its impact on breast cancer is controversial and poorly defined. In this preclinical study, we show that E4 acts as a weak estrogen by stimulating the growth of hormone-dependent breast cancer only at concentrations exceeding menopausal therapeutic needs. E4 presents also an antitumor activity by decreasing the strong proliferative effect of estradiol (E2). While estrogen receptor alpha (ERα) is the predominant receptor mediating its effects, the dual weak-estrogenic/anti-estrogenic feature of E4 results from differential signaling pathways activation. Both nuclear and rapid extra-nuclear signaling pathway are necessary for a complete estrogenic effect of E4. However, the antitumor action of E4 is not due to a capacity to antagonize E2-induced nuclear activity. Altogether, our results highlight that E4 has a limited impact on breast cancer and may offer a safe therapeutic window for the treatment of menopausal symptoms.This work was supported by grants from the Fonds de la Recherche Scientifique - FNRS (F.R.S.-FNRS, Belgium) : FRSM 3.4557.12, FRSM 3.4567.11, Télévie 7.4524.11, Télévie 7.4604.13 ; the Fonds spéciaux de la Recherche (University of Liège) : FSRC- 12/64, FSRC-12/92, FSRC-14/89, FSRC-14/65, FSRC-14/109, FSRC-14/62 ; the Centre Anticancéreux près l’ Université de Liège, the Fonds Léon Fredericq (University of Liège), the Direction Générale Opérationnelle de l’Economie, de l’Emploi et de la Recherche from the Service Public de Wallonie (DGO6, SPW, Belgium) ; the Interuniversity Attraction Poles Programme - Belgian Science Policy (Brussels, Belgium) : IAP Phase VII - P7/03 ; the Plan National Cancer (Service Public Fédéral) and the Actions de Recherche Concertées (University of Liege, Belgium): A.R.C. 11/16-02

Ontogenesis and functional aspects of oxytocin and vasopressin gene expression in the thymus network

Ontogenesis of oxytocin (OT) and vasopressin (VP) gene expression and function were investigated in murine thymus. OT and VP transcripts were detected in the thymus on embryonic days 13 and 15, respectively. Corresponding messenger RNAs were evidenced in thymic epithelial cells by in situ hybridization with a neurophysin probe. From all OT and VP receptors, only OTR was expressed by all T-cell subsets, while V1bR was found in double positive and single positive CD8 cells. In fetal thymic organ cultures, OTR antagonist d[DTyr(Et)(2), Thr(4)]OVT increased early apoptosis of CD8 cells, while V1bR antagonist (Sanofi SSR 149415) inhibited T-cell differentiation, and favored CD8 T-cell commitment. (C) 2004 Elsevier B.V. All rights reserved