AsKC11, a Kunitz Peptide from Anemonia sulcata,Is a Novel Activator of G Protein-Coupled Inward-Rectifier Potassium Channels

(1) Background: G protein-coupled inward-rectifier potassium (GIRK) channels, especially neuronal GIRK1/2 channels, have been the focus of intense research interest for developing drugs against brain diseases. In this context, venom peptides that selectively activate GIRK channels can be seen as a new source for drug development. Here, we report on the identification and electrophysiological characterization of a novel activator of GIRK1/2 channels, AsKC11, found in the venom of the sea anemone Anemonia sulcata. (2) Methods: AsKC11 was purified from the sea anemone venom by reverse-phase chromatography and the sequence was identified by mass spectrometry. Using the two-electrode voltage-clamp technique, the activity of AsKC11 on GIRK1/2 channels was studied and its selectivity for other potassium channels was investigated. (3) Results: AsKC11, a Kunitz peptide found in the venom of A. sulcata, is the first peptide shown to directly activate neuronal GIRK1/2 channels independent from Gi/o protein activity, without affecting the inward-rectifier potassium channel (IRK1) and with only a minor effect on K_V1.6 channels. Thus, AsKC11 is a novel activator of GIRK channels resulting in larger K⁺ currents because of an increased chord conductance. (4) Conclusions: These discoveries provide new insights into a novel class of GIRK activators.     

Influence of dietary carbohydrate on antioxidant enzyme activities in liver of Atlantic salmon (Salmo salar L.)

Effects of dietary carbohydrates on hepatic antioxidant enzymeactivities were studied in Atlantic salmon (Salmo salarL.), fed diets containing no additional carbohydrate, or a low (15% addition)orhigh (30% addition) concentration of D-glucose or gelatinised potato starch.Addition of free glucose to the feeds resulted in glucose availability being 7and 18% when added at 15 and 30%, while the addition of gelatinised starch didnotseem to reduce starch availability. The dietary treatment groups (nocarbohydrate, 7 and 18% glucose or 15 and 30% gelatinised starch) were providedwith quantities of feed that supplied the same amounts of protein and lipid.There was a positive correlation between tiobarbituric acid-reactive substances(TBARS) in feeds and salmon liver. Liver glycogen concentrations increased withincreasing dietary glucose and starch, and the two higher levels of liverglycogen, found in groups G18 and S30, resulted in decreased activities ofcatalase, superoxide dismutase (t-SOD), and concentrations of glutathione. Thisindicates that there are links between carbohydrate metabolism and antioxidantsystems in salmon liver. There was no correlation between selenium dependentglutathione peroxidase (Se-GPX) activity and dietary carbohydrateconcentrations, but Se-GPX activity increased in response to increased TBARS.Catalase and t-SOD activities did not correlate with either TBARS values orSe-GPX activity.     

Immune response and disease resistance in Atlantic salmon (Salmo salar L.) fed three levels of dietary vitamin E and the effect of vaccination on the liver status of antioxidant vitamins

Atlantic salmon were fed 40, 300 or 1100 mgall-rac--tocopheryl acetate kg feed^–1 for 12 weeks.After 6 weeks half of the fish in each group were marked and vaccinated (i.p.injection) against furunculosis and vibriosis. Liver -tocopherol levelsreflected the dietary input after 6 and 12 weeks of feeding. Noimmunomodulatoryeffects of dietary vitamin E on baseline levels of the immune parameters beforevaccination were detected in this study, as evaluated by antibody dependent andspontaneous plasma complement activities. In general, vaccination increasedplasma complement activities and the number of antigen specific antibodyproducing cells as compared with unvaccinated control fish, but with nodifferences with respect to the vitamin E regimes. Also the ability ofunvaccinated fish to withstand experimental furunculosis was unaffected bydietary vitamin E. The concentrations of -tocopherol and ascorbic acidinthe liver were, however, negatively affected 6 weeks post vaccination comparedto respective unvaccinated fish.     

The Applicability of Agar-Diffusion Test in Measuring Deoxyribonucleases in Pig Intestinal Content

The suitability of an agar-diffusion test (ADT) using toluidine blue deoxyribonucleic acid agar (TDA) for measuring DNase activity in pig intestinal contents was investigated. The ADT was compared with a spectrophotometrical method. Distinct metachromatic zones around wells in the DNA-containing agar, into which the intestinal content was applied, indicated DNase activity. The DNase activity was determined semiquantitatively by making serial twofold dilutions of the intestinal content. The spectrophotometrical method was optimal at pH 7.2. The ADT proved to be most sensitive at pH 5.6. The ability of the 2 methods employed to measure low concentrations of DNases was equal. However, the ADT was considered more suitable than the spectrophotometrical method because ADT measured reduced amounts of enzyme. DNase activity was demonstrated throughout the small intestine and in the large intestine. By the zymogram technique, at least 3 different DNases could be demonstrated in the lower parts of the small intestine, 1 of which could be of extrapancreatic origin.     

Effect of induced hyperoxia on the antioxidant status of Atlantic salmon Salmo salar L. fed three different levels of dietary vitamin E

Atlantic salmon smolt (64 g) were fed 40, 300 or 1100 mg kg^?1 all‐rac‐α‐tocopheryl acetate feed under normoxic and moderate hyperoxic conditions for 12 weeks. After 7 weeks, however, the hyperoxic fish were subjected to a temporary severe supersaturation of dissolved oxygen with development of gas bubble disease and substantial mortality as a result. Liver α‐tocopherol (α‐TOH) levels reflected the dietary input after 6 and 12 weeks of feeding. Increased levels of thiobarbituric‐reactive substances (TBARS) in the liver of hyperoxygenated fish after 6 weeks indicate that moderate hyperoxic conditions may represent an oxidative challenge to the fish. After 12 weeks, liver ascorbic acid (AA) content was reduced in hyperoxygenated fish, and a decreasing tendency was also found in the liver content of α‐TOH. Effects of oxygen supersaturation were also observed on liver antioxidant enzyme activities, content of glutathione and total mercapthans in liver, phagocytic chemiluminescent response and total plasma protein level, indicating a disturbed redox balance. No effects of dietary vitamin E were observed on any of the measured parameters, indicating that high levels of dietary vitamin E under these circumstances did not protect the fish against oxidative stress.     

Effects of Oscillatoria agardhii-toxins on blood pressure and isolated organ preparations

Toxic extracts of the blue-green alga Oscillatoria agardhii were tested for their effect on various isolated laboratory animal preparations. The Oscillatoria-toxins did not show hemagglutinative or hemolytical properties. An immediate rise in blood pressure was seen after intravenous injection in rats. As regards isolated organs, effects were most marked on the liver. Addition of toxic extract to isolated liver perfusion fluid led to physiological, biochemical and pathological changes indicating severe damage to the organ. Negative chronotropic and inotropic effects were found on the isolated heart. The toxins did not influence isolated smooth muscle or nerve striated muscle preparations.     

Roe deer (Capreolus capreolus) browsing pressure affects yew (Taxus baccata) recruitment within nature reserves in Norway

Grazing and browsing by large herbivores is known to affect patterns of biodiversity. Still much remains to be done to assess the specific role of browsing on single, red listed plant species. In Norway, the yew (Taxus baccata) is a threatened species and several nature reserves have been established. We show, that within two nature reserves between 1988 and 2003, yew recruitment was strongly dependent on variation in roe deer (Capreolus capreolus) browsing pressure. Our study emphasized that establishing nature reserves for yew is not enough to conserve this species unless control over browsing roe deer population is also taken. We argue that roe deer, being the most abundant large herbivore in Europe, may have a larger effect on conservation than currently realized.     

Fimbriae in Escherichia coli isolated from the small intestine of piglets

Ninety E. coli strains, isolated from piglets which had died from neonatal diarrhea, were tested for the presence of K88, K99, 987P and type 1 fimbriae. Two or more types of fimbriae were demonstrated in 14 of the strains, a single fimbria! type in 44 strains while in 32 strains no fimbriae were detected. Of the 14 E. coli strains with more than 1 type of fimbriae, 12;, 10, 8 and 4 strains showed K88, K99, 987P and type 1, respectively.Twelve E. coli strains were isolated from piglets which had died in the neonatal period without showing signs of neonatal diarrhea at necropsy. One strain showed 987P and 3 strains showed type 1 fimbriae, while the remaining 8 strains were unfimbriated.Sixteen fimbriated E. coli strains were subcultured in order to examine the stability of fimbrial expression in the strains. The K88 and the type 1 fimbriae were regularly expressed, while the K99 and 987P were inconsistently demonstrated.