The molecular pathogenesis of infectious spleen and kidney necrosis virus (ISKNV) infections is important but has rarely been studied in connection to host organelle behavior. In the present study, we demonstrated that ISKNV can induce host cell death via a pro-apoptotic Bcl-2 and anti-apoptotic Bcl-2 family member imbalance in mitochondrial membrane potential (MMP or ΔΨm) regulation in GF-1 cells. The results of our study on ISKNV infection showed that it can induce host cell death by up to 80% at day 5 post-infection. Subsequently, in an apoptotic assay, ISKNV infection was seen to induce an increase in Annexin-V-positive signals by 20% and in propidium iodide (PI) staining-positive signals by up to 30% at day 5 (D5) in GF-1 cells. Then, through our studies on the mechanism of cell death in mitochondria function, we found that ISKNV can induce MMP loss by up to 58% and 78% at days 4 and 5 with a JC1 dye staining assay. Furthermore, we found that pro-apoptotic members Bax and Bak were upregulated from the early replication stage (day one) to the late stage (day 5), but the expression profiles were very dynamically different. On the other hand, by Western blotted analysis, the anti-apoptotic members Bcl-2 and Bcl-xL were upregulated very quickly at the same time from day one (two-fold) and continued to maintain this level at day five. Finally, we found that pro-apoptotic death signals strongly activated the downstream signals of caspase-9 and -3. Taken together, these results suggest that ISKNV infection can induce Bax/Bak-mediated cell death signaling downstream of caspase-9 and -3 activation. During the viral replication cycle with the cell death induction process, the anti-apoptotic members Bcl-2/Bcl-xL interacted with the pro-apoptotic members Bax/Bak to maintain the mitochondrial function in the dynamic interaction so as to maintain the MMP in GF-1 cells. These findings may provide insights into DNA-virus control and treatment.

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Infectious spleen and kidney necrosis virus (ISKNV) infections can trigger host cell death and are correlated with viral replication; however, they have rarely been considered in terms of the host organelle involvement. In the present study, we demonstrated that ISKNV triggered an oxidative stress signal in the Nrf2-mediated oxidative stress response and induced stress signals for Bax/Bak-mediated host cell death in fish GF-1 cells. The results showed that after ISKNV infection, the levels of reactive oxidative species (ROS) increased by 60–80% from day 3 to day 5, as assessed by an H2DCFDA assay for tracing hydrogen peroxide (H2O2), which was correlated with up to a one-fold change in the fish GF-1 cells. Furthermore, we found that ISKNV infection induced Nrf2-mediated ROS stress signals from D1 to D5, which were correlated with the upregulation of antioxidant enzymes, such as catalase, SOD1, and SOD2; these effects were blocked by the antioxidants GSH and NAC. By analyzing Nrf2-mediated ROS stress signals for cell death regulation via an apoptotic assay, we found that treatment with antioxidants reduced annexin-V-positive signals by 10% (GSH) to 15% (NAC); moreover, necrotic-positive signals were reduced by 6% (GSH) and 32% (NAC) at day 5 (D5) in GF-1 cells, as indicated by PI staining. Furthermore, we found that Nrf2-mediated ROS stress regulated mitochondrion-mediated Bax/Bak death signals at D3 and D5; this was effectively blocked by antioxidant treatment in the GF-1 cells, as demonstrated by a JC1 assay (ΔΨm) and western blot analysis. In addition, we found that downstream signals for caspase-9 and -3 activation were apparently blocked by antioxidant treatment at D3 and D5. Finally, we found that treatment with GSH and NAC reduced major capsid protein (MCP) expression and virus titer (TCID50%) by up to 15-fold at D5 in GF-1 cells. Thus, our data suggest that ISKNV can induce ROS production, which triggers Nrf2-mediated stress signals. Then, these stress signals can regulate mitochondrion-mediated Bax/Bak apoptotic signaling, which is connected to downstream caspase-9 and -3 activation. If ISKNV-induced Nrf2-mediated stress signaling is blocked, then the antioxidants GSH and NAC can also suppress apoptotic signals or reduce viral replication. These findings may provide insights into the control and treatment of double-stranded DNA viruses.

By organizing the genome into gene modules (GMs), a living cell coordinates the activities of a set of genes to properly respond to environmental changes. The transcriptional regulation of the expression of a GM is usually carried out by a cooperative transcription factor set (CoopTFS) consisting of several cooperative transcription factors (TFs). Therefore, a database which provides CoopTFSs and their target GMs is useful for studying the cellular responses to internal or external stimuli. To address this need, here we constructed YGMD (Yeast Gene Module Database) to provide 34120 CoopTFSs, each of which consists of two to five cooperative TFs, and their target GMs. The cooperativity between TFs in a CoopTFS is suggested by physical/genetic interaction evidence or/and predicted by existing algorithms. The target GM regulated by a CoopTFS is defined as the common target genes of all the TFs in that CoopTFS. The regulatory association between any TF in a CoopTFS and any gene in the target GM is supported by experimental evidence in the literature. In YGMD, users can (i) search the GM regulated by a specific CoopTFS of interest or (ii) search all possible CoopTFSs whose target GMs contain a specific gene of interest. The biological relevance of YGMD is shown by a case study which demonstrates that YGMD can provide a GM enriched with genes known to be regulated by the query CoopTFS (Cbf1-Met4-Met32). We believe that YGMD provides a valuable resource for yeast biologists to study the transcriptional regulation of GMs. Database URL: http://cosbi4.ee.ncku.edu.tw/YGMD/, http://cosbi5.ee.ncku.edu.tw/YGMD/ or http://cosbi.ee.ncku.edu.tw/YGMD/

Because carbon dioxide adsorbs the radiation from the Sun and the Earth’s surface, global warming has become a severe problem in this century. Global warming causes many environmental problems such as heatwave, desertification, and erratic rainfall. Above all, erratic rainfall makes people have insufficient freshwater. To solve this problem, desalination technology has been developed in many countries. Although desalination technology can provide freshwater, it produces brine as well (producing 1 L of freshwater would result in 1 L of brine). The brine will decrease the dissolved oxygen in the sea and affect the organism’s habitat. In this study, magnesium and calcium from desalination brine were recovered in the form of magnesium hydroxide and calcium hydroxide by adjusting the pH value for carbon capture and sodium removal. Magnesium hydroxide would turn into magnesium carbonate through contacting CO2 in saturated amine carriers. Calcium hydroxide was added to the brine and reacted with CO2 (modified Solvay process). Sodium in brine would then be precipitated in the form of sodium bicarbonate. After removing sodium, brine can be released back into the ocean, or other valuable metals can be extracted from brine without the side effect of sodium. The results revealed that 288 K of 3-Amino-1-propanol could capture 15 L (26.9 g) of CO2 and that 25 g/L of Ca(OH)2 at 288 K was the optimal parameter to remove 7000 ppm sodium and adsorb 16 L (28.7 g) of CO2 in the modified Solvay process. In a nutshell, this research aims to simultaneously treat the issue of CO2 emission and desalination brine by combining the amines carrier method and the modified Solvay process.