Tagging of Litopenaeus vannamei (Boone) by uropod cutting: a comparison of two methods

Two uropod‐cutting methods were used and compared as a means of long‐term tagging on juvenile (mean weight: 1.77±0.5 g) and sub‐adult (mean weight: 18.7±0.2 g) Litopenaeus vannamei: (a) cutting half of the left exopod (LE); and (b) cutting the whole right uropods (WR). Shrimp with uncut uropods served as controls. Regeneration of uropods in juvenile shrimp commenced within a week and complete regeneration was evident after 2 weeks. In sub‐adult shrimps, incomplete regeneration (IG) was observed within 1 month of culture in both cutting methods. Complete regeneration was observed in 2 months among shrimps in LE but not in WR. Abnormal growths (AGs) on regenerated uropods (91.1%) were observed in juvenile shrimp including hollow‐type (HT; 77.8%) and flat‐type (FT; 13.3%). Among sub‐adult shrimp, 87.6% produced AG including HT (68.8%) and FT (18.8%). In WR, regenerated uropods among juvenile shrimp were of a normal type (N) which was indistinguishable from the control treatment. On the other hand, normal growth (with less pigmentation, 5.2%), IG (26.3%), and AG (68.5%) were observed among sub‐adult shrimp. After 4 months of culture, with average final weights of 18–19 g, tag retention rate in juvenile shrimps was 91.1% for LE treatment, and 87.6% (LE) and 100% (WR) in sub‐adults. Survival rate was about 98% for both cutting methods in juvenile and 53–63% for sub‐adult.     

Proteomic Investigation of the Sinulariolide-Treated Melanoma Cells A375: Effects on the Cell Apoptosis through Mitochondrial-Related Pathway and Activation of Caspase Cascade

Sinulariolide is an active compound isolated from the cultured soft coral Sinularia flexibilis. In this study, we investigated the effects of sinulariolide on A375 melanoma cell growth and protein expression. Sinulariolide suppressed the proliferation and migration of melanoma cells in a concentration-dependent manner and was found to induce both early and late apoptosis by flow cytometric analysis. Comparative proteomic analysis was conducted to investigate the effects of sinulariolide at the molecular level by comparison between the protein profiles of melanoma cells treated with sinulariolide and those without treatment. Two-dimensional gel electrophoresis (2-DE) master maps of control and treated A375 cells were generated by analysis with PDQuest software. Comparison between these maps showed up- and downregulation of 21 proteins, seven of which were upregulated and 14 were downregulated. The proteomics studies described here identify some proteins that are involved in mitochondrial dysfunction and apoptosis-associated proteins, including heat shock protein 60, heat shock protein beta-1, ubiquinol cytochrome c reductase complex core protein 1, isocitrate dehydrogenase (NAD) subunit alpha (down-regulated), and prohibitin (up-regulated), in A375 melanoma cells exposed to sinulariolide. Sinulariolide-induced apoptosis is relevant to mitochondrial-mediated apoptosis via caspase-dependent pathways, elucidated by the loss of mitochondrial membrane potential, release of cytochrome c, and activation of Bax, Bad and caspase-3/-9, as well as suppression of p-Bad, Bcl-xL and Bcl-2. Taken together, our results show that sinulariolide-induced apoptosis might be related to activation of the caspase cascade and mitochondria dysfunction pathways. Our results suggest that sinulariolide merits further evaluation as a chemotherapeutic agent for human melanoma.     

Induction of Apoptosis by 11-Dehydrosinulariolide via Mitochondrial Dysregulation and ER Stress Pathways in Human Melanoma Cells

In this study the isolated compound 11-dehydrosinulariolide from soft coral Sinularia leptoclados possessed anti-proliferative, anti-migratory and apoptosis-inducing activities against A2058 melanoma cells. Anti-tumor effects of 11-dehydrosinulariolide were determined by MTT assay, cell migration assay and flow cytometry. Growth and migration of melanoma cells were dose-dependently inhibited by 2–8 μg/mL 11-dehydrosinulariolide. Flow cytometric data indicated that 11-dehydrosinulariolide induces both early and late apoptosis in melanoma cells. It was found that the apoptosis induced by 11-dehydrosinulariolide is relevant to mitochondrial-mediated apoptosis via caspase-dependent pathways, elucidated by loss of mitochondrial membrane potential (∆Ψm), release of cytochrome C, activation of caspase-3/-9 and Bax as well as suppression of Bcl-2/Bcl-xL. The cleavage of PARP-1 suggested partial involvement of caspase-independent pathways. Immunoblotting data displayed up-regulations of PERK/eIF2α/ATF4/CHOP and ATF6/CHOP coupling with elevation of ER stress chaperones GRP78, GRP94, calnexin, calreticulin and PDI, implicating the involvement of these factors in ER stress-mediated apoptosis induced by 11-dehydrosinulariolide. The abolishment of apoptotic events after pre-treatment with salubrinal indicated that ER stress-mediated apoptosis is also induced by 11-dehydrosinulariolide against melanoma cells. The data in this study suggest that 11-dehydrosinulariolide potentially induces apoptosis against melanoma cells via mitochondrial dysregulation and ER stress pathways.     

Omega-3 Fatty Acid Intervention Suppresses Lipopolysaccharide-Induced Inflammation and Weight Loss in Mice

Bacterial endotoxin lipopolysaccharide (LPS)-induced sepsis is a critical medical condition, characterized by a severe systemic inflammation and rapid loss of muscle mass. Preventive and therapeutic strategies for this complex disease are still lacking. Here, we evaluated the effect of omega-3 (n-3) polyunsaturated fatty acid (PUFA) intervention on LPS-challenged mice with respect to inflammation, body weight and the expression of Toll-like receptor 4 (TLR4) pathway components. LPS administration induced a dramatic loss of body weight within two days. Treatment with n-3 PUFA not only stopped loss of body weight but also gradually reversed it back to baseline levels within one week. Accordingly, the animals treated with n-3 PUFA exhibited markedly lower levels of inflammatory cytokines or markers in plasma and tissues, as well as down-regulation of TLR4 pathway components compared to animals without n-3 PUFA treatment or those treated with omega-6 PUFA. Our data demonstrate that n-3 PUFA intervention can suppress LPS-induced inflammation and weight loss via, at least in part, down-regulation of pro-inflammatory targets of the TLR4 signaling pathway, and highlight the therapeutic potential of n-3 PUFA in the management of sepsis.     

13C-NMR spectroscopy studies of humic substances in subtropical perhumid montane forest soil

We investigated soil organic matter in a forest of natural Hinoki cypress (Chamaecyparis obtusa) under perhumid weather conditions in north central Taiwan. Humic substances along the transect from the summit and footslope to lakeshore were characterized by use of solid-state cross-polarization, magic-angle-spinning ¹³C nuclear magnetic resonance spectroscopy (CP/MAS ¹³C-NMR). The major components of soil organic carbon in whole soil and humic substances were alkyl-C, O-alkyl-C, and di-O-alkyl-C, ranging from 60.6% to 80.7%, then aromatic-C, 7.5% to 9.8%. The degree of humification of soil organic matter, both O-alkyl-C/alkyl-C ratio and aromaticity, decreased slightly from the summit to lakeshore. The content of functional groups of polar and acidic groups, including O-alkyl-C, di-O-alkyl-C, and carboxyl-C, corresponded with the topographical effect, increasing slightly from the summit to lakeshore. However, the relatively low degree of humification in soils of this perhumid forest and low aromaticity were due to high precipitation and acidity, which appeared to hinder organic matter decomposition with topography change.     

Degradation of Di-(2-ethylhexyl) Phthalate (DEHP) by TiO2 Photocatalysis

The photocatalytic degradation of di-(2-ethylhexyl) phthalate (DEHP) in solution using titanium dioxide (TiO₂) was analyzed in this study. It was found that DEHP was completely removed in the solution after 150 min irradiation. The effect of different factors, such as photocatalyst amount, DEHP concentration, light intensity, pH, and temperature on photocatalytic degradation was investigated. The degradation mechanism of DEHP with proton and hydroxyl radicals oxidation were also studied. It is suggested that either ethylhexyl or ester chain scissions of the aliphatic part of DEHP was the dominant degradation mechanism of the process. The photocatalytic degradation process was well described by first-order reaction. The final mineralization product was carbon dioxide and the intermediate products were identified by GC-MS. Thus, the photocatalytic degradation treatment of DEHP in wastewater is a relative simple and fast method.     

Photocatalytic Degradation of Azo Dye Reactive Orange 16 by TiO2

Water, Air, & Soil Pollution - Several recent reports document increasing concentrations of chloride in surface waters of northeastern and midwestern North America. These patterns, together...