Four tropical marine fish cell lines have been established from the eye, fin, heart and swim bladder of grouper, Epinephelus awoara (Temminck & Schlegel). Optimum media and temperature conditions for maximum growth were standardized. The eye and swim bladder cells were mostly epithelial, but the fin and heart cells were mostly fibroblastic. The viability of cells was 95% after 1 year of storage in liquid nitrogen (-196 degrees C). Besides these four cell lines, previously established grouper brain, kidney and liver cell lines were also used for a viral susceptibility study which showed that all the cell lines were sensitive to grouper iridovirus, whereas only brain, fin and liver cell lines were susceptible to the yellow grouper nervous necrosis virus (a nodavirus). Electron microscopy studies of the grouper irido- and nodaviruses in ultrathin sections of infected cells showed an abundance of viral particles in the cytoplasm of the virus-infected cells indicating the effective replication of these two viruses. It is suggested that these cell lines can be used for the isolation of putative fish specific viruses and provide a valuable tool to study the mechanisms of host-pathogen interactions. Furthermore, these cell lines upon transfection, using pEGFP-C1 and pEGFP-aMT2.5 (ayu metallothionein promoter), produced significant fluorescent signals indicating their utility for exogenous studies.

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A number of prior studies have developed a variety of multivariate volatility models to describe the joint distribution of spot and futures, and have applied the results to form the optimal futures hedge. In this study, the authors propose a new class of multivariate volatility models encompassing realized volatility (RV) estimates to estimate the risk-minimizing hedge ratio, and compare the hedging performance of the proposed models with those generated by return-based models. In an out-of-sample context with a daily rebalancing approach, based on an extensive set of statistical and economic performance measures, the empirical results show that improvement can be substantial when switching from daily to intraday. This essentially comes from the advantage that the intraday-based RV potentially can provide more accurate daily covariance matrix estimates than RV utilizing daily prices. Finally, this study also analyzes the effect of hedge horizon on hedge ratio and hedging effectiveness for both the in-sample and the out-of-sample data.

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In this study, the phenomenon of light trapping in Si solar cells coated with metal (Au) and dielectric (TiO 2 , SiO 2 ) nanoparticles (NPs) is systematically investigated. In contrast to previous reports, herein it is proposed that the photocurrent enhancement of solar cells should be attributed to the limited antirefl ection ability of the Au NP arrays. In other words, the Au NP arrays might not enhance the absorption of the active layer in cells when no light is refl ected from the air-substrate interface. Therefore, the Au NPs are replaced with dielectric NPs, which possess lower extinction coeffi cients, and then the antirefl ection property of the TiO 2 NP arrays is optimized. A simple, rapid, and cheap solution-based method is used to prepare close-packed TiO 2 NP fi lms on Si solar cells; these devices exhibit a uniform and remarkable increase (ca. 30%) in their photocurrents. To the best of the authors' knowledge, this uniform photocurrent enhancement is greater than those obtained from previously reported metal and dielectric NP-enhanced Si wafer-based solar cells.

Two new cell lines have been established from the muscle and swim bladder tissues of barramundi, Lates calcarifer, and designated as BM (barramundi muscle) and BSB (barramundi swimbladder), respectively. The cells multiplied well at 28 degrees C in Leibovitz's L-15 medium supplemented with 10% foetal bovine serum, and have been continuously subcultured more than 100 times to date. Morphologically, BM cells were mostly fibroblastic, whereas BSB were mostly epithelial. Both cell lines were susceptible to grouper iridovirus (GIV) and displayed characteristics of apoptosis after viral infection. The induction of apoptosis was further assayed in GIV-infected BM and BSB cells by various methods. The inhibition of cell growth by GIV was demonstrated by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. Morphological observations revealed typical apoptotic features in the infected cells, including cell shrinkage and rounding, chromosome condensation and formation of apoptotic body-like vesicles. Chromosome fragmentation was detected by DNA laddering and TUNEL assays. Finally, the appearance of phosphotidylserine on the outer leaflet of apoptotic cell membranes was confirmed by annexin V staining. This is the first report of apoptosis induced by GIV in fish cells.

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BackgroundGrouper (Epinephelus spp) is an economically important fish species worldwide. However, viral pathogens such as nervous necrosis virus (NNV) have been causing severe infections in the fish, resulting in great loss in the grouper aquaculture industry. Yet, the understanding of the molecular mechanisms underlying the pathogenicity of NNV is still inadequate, mainly due to insufficient genomic information of the host.ResultsDe novo assembly of grouper transcriptome in the grouper kidney (GK) cells was conducted by using short read sequencing technology of Solexa/Illumina. A sum of 66,582 unigenes with mean length of 603 bp were obtained, and were annotated according to Gene Ontology (GO) and Clusters of Orthologous Groups (COG). In addition, the tag-based digital gene expression (DGE) system was used to investigate the gene expression and pathways associated with NNV infection in GK cells. The analysis revealed endoplasmic reticulum (ER) stress response was prominently affected in NNV-infected GK cells. A further analysis revealed an interaction between the NNV capsid protein and the ER chaperone immunoglobulin heavy-chain binding protein (BiP). Furthermore, exogenous expression of NNV capsid protein was able to induce XBP-1 mRNA splicing in vivo, suggesting a role of the capsid protein in the NNV-induced ER stress.ConclusionsOur data presents valuable genetic information for Epinephelus spp., which will benefit future study in this non-model but economically important species. The DGE profile of ER stress response in NNV-infected cells provides information of many important components associated with the protein processing in ER. Specifically, we showed that the viral capsid protein might play an important role in the ER stress response.