Systemic and strict regulation of the glutathione redox state in mitochondria and cytosol is needed for zebrafish ontogeny.

BACKGROUND: Redox control seems to be indispensable for proper embryonic development. The ratio between glutathione (GSH) and its oxidized disulfide (GSSG) is the most abundant cellular redox circuit. METHODS: We used zebrafish harboring the glutaredoxin 1-redox sensitive green fluorescent protein (Grx1-roGFP) probe either in mitochondria or cytosol to test the hypothesis that the GSH:GSSG ratio is strictly regulated through zebrafish embryogenesis to sustain the different developmental processes of the embryo. RESULTS: Following the GSSG:GSH ratio as a proxy for the GSH-dependent reduction potential (EhGSH) revealed increasing mitochondrial and cytosolic EhGSH during cleavage and gastrulation. During organogenesis, cytosolic EhGSH decreased, while that of mitochondria remained high. The similarity between EhGSH in brain and muscle suggests a central regulation. Modulation of GSH metabolism had only modest effects on the GSSG:GSH ratios of newly hatched larvae. However, inhibition of GSH reductase directly after fertilization led to dead embryos already 10 h later. Exposure to the emerging environmental pollutant Perfluorooctane Sulfonate (PFOS) disturbed the apparent regulated EhGSH as well. CONCLUSIONS: Mitochondrial and cytosolic GSSG:GSH ratios are almost identical in different organs during zebrafish development indicating that the EhGSH might follow H2O2 levels and rather indirectly affect specific enzymatic activities needed for proper embryogenesis. GENERAL SIGNIFICANCE: Our data confirm that vertebrate embryogenesis depends on strictly regulated redox homeostasis. Disturbance of the GSSG:GSH circuit, e.g. induced by environmental pollution, leads to malformation and death.

RNA form baker's yeast cultured with and without lipopolysaccharide (LPS) modulates gene transcription in an intestinal epithelial cell model, RTgutGC from rainbow trout (Oncorhynchus mykiss).

The objective of this study was to evaluate if the intestinal RTgutGC cell line could be suitable for research on dietary ingredients and their function as modulators of inflammation during lipopolysaccharide (LPS) induced stress. The RTgutGC cells cultured together with RNA from baker's yeast, reached confluency after 72 h. The cells were grown in either compete L-15 (CM) or nutrient deprived L-15 (DM). Then, the RTgutGC cells were exposed to LPS or RNA from baker's yeast, either alone, or in combination, in CM or DM. All cultures were harvested following LPS challenge for 48 h and 72 h. LPS induced transcription of Interleukin 1ß (IL-1ß), Interleukin -8 (IL-8), Toll like receptor 3 (TLR3), interferon regulating factor 3 (irf3), Nuclear factor Ä¸ß (NFĸß), one of the multidrug transporters, ABCC2, and glutamine synthase 1 (GLS01) in RTgutGC cells at one or both sampling points (48 h and/or 72 h post LPS challenge). RNA from baker's yeast in culture alone, (cultured 120 h and 144 h with RTgutGC cells and harvested at the respective LPS sampling points) induced transcription of INF1, TNFα and ticam/trif, not induced by LPS. In addition, RNA from baker's yeast affected IL-1ß, TLR3, irf3 and NFĸß, comparable to the responses triggered by LPS. RNA from baker's yeast alone did not affect ABCC2 or GLS01 transcriptions in this set up. So, LPS and RNA from baker's yeast affects distinct but also common gene transcripts in this intestinal cell line. Culturing RTgutGC cells in DM, adding a combination of LPS and RNA from baker's yeast, reduced IL-1ß transcription compared to cells grown in CM, 48 h and 72 h post LPS challenge. Also, in RTgutGC cells, grown in DM, the LPS induced transcription of ABCC2 declined, measured 48 h post LPS challenge. Possibly indicating that optimal transcription of IL-1ß and ABBC2 in RTgutGC cells, cultured over time, requires access of adequate nutrients under stressful condition. RNA from baker's yeast induced INF1 transcription in the RTgutGC cells, regardless if the medium was complete or deprived of nutrients. However, culturing RTgutGC cells in DM enriched with RNA from baker's yeast for a longer period of time (120 h, 144 h), seemed beneficial for INF1 transcription.

Daily Intake of Protein from Cod Residual Material Lowers Serum Concentrations of Nonesterified Fatty Acids in Overweight Healthy Adults: A Randomized Double-Blind Pilot Study.

Improved process technologies have allowed fishing vessels to utilize residuals from cod fillet production (head, backbone, skin, cuttings, and entrails) and convert this to high-quality protein powders for human consumption. In this double-blind pilot study, 42 healthy overweight or obese adults were randomized to three experimental groups consuming tablets corresponding to 6 g/day of proteins from cod residuals as presscake meal (Cod-PC), presscake and stickwater meal (Cod-PCW), or placebo tablets (control) for eight weeks. The primary outcome of this study was changes in metabolites related to glucose regulation in overweight or obese healthy adults after intake of proteins from cod residuals. Cod-PC supplementation decreased postprandial serum nonesterified fatty acids (NEFA) concentration and increased gene expressions of diglyceride acyltransferase 1 and 2 in subcutaneous adipose tissue compared with controls. Fasting insulin increased while fasting NEFA and 120-min postprandial glucose decreased within the Cod-PC group, but these changes did not differ from the other groups. In conclusion, supplementation with Cod-PC beneficially affected postprandial serum NEFA concentration compared with the other groups in overweight or obese adults. Supplementation with Cod-PCW, which contains a higher fraction of water-soluble protein compared to Cod-PC, did not affect serum markers of glucose regulation.