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Open reading frame 89 (ORF89) is one of the three genes that are believed to be involved in the latent infection of white spot syndrome virus (WSSV). Here, we report the structure and functional characterization of ORF89. cDNA sequencing, 5′ RLM-RACE, and 3′ RLM-RACE showed that ORF89 gene is transcribed into an unspliced mRNA of 4436 nucleotides, which is predicted to encode a protein of 1437 amino acids. ORF89 expressed an approximately 165-kDa protein in Sf9 cells that localized in the nucleus. Amino acids 678–683 were found to be essential for nuclear localization. Cotransfection assays demonstrated that ORF89 protein repressed its own promoter as well as those of a protein kinase and the thymidine–thymidylate kinase genes of WSSV. SYBR Green real-time PCR indicated that the repression occurred at the transcriptional level. [Copyright &y& Elsevier]

The mud lobster, genus Thalassina is the least studied taxon compared to other mud crabs within Thalassinidae family. Burrowing thalassinid populations are considered to be an effective means of bioturbation, affecting the macro- and micronutrient properties in the substrates they occur in through nutrient redistribution. This study assessed the combined effects of fresh mud excavation and old mud deposition on soil turnover and vertical marconutrients including NH 4 –N, NO 3 –N, P, K, S, Na, Ca, and Mg and micronutrients including Fe, Cu, Zn and Mn distributions in a Malaysian mangrove, Sibuti Wildlife Sanctuary, Sarawak. The mud lobster Thalassina anomala (Decapoda: Thalassinidae) inhabits and builds mounds both seafront and inland mangroves. Their burrowing activities increased NH 4 –N, NO 3 –N, P, and Na concentrations in fresh mud and Fe, Cu and Zn concentrations in the old mud. The nutrient concentrations in excavated soils were greatly affected by habitat type, and depth. Burrowing processes can effectively turnover subsurface soil to the surface, which acidifies strongly upon oxidation. Higher S content in mud lobster soil together with low pH revealed mud lobsters play a strong role in the development of acidity in the study areas. These processes could stimulate formation of acid sulphate soil in the mangroves. Therefore, bioturbation not only affects soil physical and chemical properties but also ecological functions of mangroves, and needs proper attention during restoration and reclamation programs of mangrove forests. • Effects of mud lobster mound on mangrove soil nutrient turnover are reported for the first time. • Microclimate of the mound occurring in two mangroves under this study is distinctive. • Burrowing activities play a strong role in the development of acidity in the study areas. • Distribution of the nutrients in the mound and mangrove soils by depth is compared. [ABSTRACT FROM AUTHOR]

Abstract: Previous studies have shown that boar sperm quality after cryopreservation differs depending on the ejaculate fraction used and that spermatozoa contained in the first 10mL (P1) of the sperm-rich fraction (SRF) show better cryosurvival than those in the SRF-P1. Since protein tyrosine phosphorylation (PTP) in spermatozoa is related with the tolerance of spermatozoa to frozen storage and cryocapacitation, we assessed the dynamics of cryopreservation-induced PTP and intracellular calcium ([Ca2+]i) in spermatozoa, using flow cytometry, from P1 and SRF-P1 of the boar ejaculate at different stages of cryopreservation. Sperm kinetics, assessed using a computer-assisted semen analyzer, did not differ between P1 and SRF-P1 during cryopreservation but the decrease in sperm velocity during cryopreservation was significant (P <0.05) in SRF-P1 compared to P1. There were no significant differences in percentages of spermatozoa with high [Ca2+]i between P1 and SRF-P1 in fresh as well as in frozen–thawed semen. A higher (P <0.001) proportion of spermatozoa displayed PTP during the course of cryopreservation indicating a definite effect of the cryopreservation process on sperm PTP. The proportion of spermatozoa with PTP did not differ significantly between portions of the boar ejaculate. However at any given step during cryopreservation the percentage of spermatozoa with PTP was comparatively higher in SRF-P1 than P1. A 32kDa tyrosine phosphorylated protein, associated with capacitation, appeared after cooling suggesting that cooling induces capacitation-like changes in boar spermatozoa. In conclusion, the study has shown that the cryopreservation process induced PTP in spermatozoa and their proportions were similar between portions of SRF. [Copyright &y& Elsevier]