Induction and sequencing of Rousette bat interferon alpha and beta genes

Bats are considered to be natural reservoirs for several viruses of clinical importance, including rabies virus, Nipah virus, and Hendra virus. Type I interferons (IFNs) is an important part of the immune system in the defense against viral infection. To investigate the function of type I IFNs upon viral infection in bats, the nucleic acid, and amino acid sequences of Egyptian Rousette (Rousettus aegyptiacus) IFN-alpha and -beta were characterized. Sequence data indicated that bat IFN-alpha consists of 562-bp encoded 187-aa, and IFN-beta consisted of 558-bp encoded 186-aa. Phylogenetic analysis of the overall identity of IFN-beta shared the highest sequence homology with pig IFN-beta in both nucleotide and amino acid level. Stimulation of bat primary kidney cells (BPKCs) and bat lung cell lines, Tb-1 Lu, with polyinosinic-polycytidylic acid (poly(I:C)) or exogenous bat type I IFNs resulted in increased type I IFNs mRNA expression in BPKCs, but not in Tb-1 Lu. Characterization of the bat IFN-alpha and -beta genes allows understanding of the immune responses upon stimulation in different tissues, thus providing practical strategies for control and treatment of clinically important diseases. These results are important especially for the virus infection, and suggest that future molecular studies on virus infection experiment of bats in vitro will require careful consideration of the differences of type I IFN expression patterns in different cell types.     

Fucoidan from Fucus vesiculosus Protects against Alcohol-Induced Liver Damage by Modulating Inflammatory Mediators in Mice and HepG2 Cells

Fucoidan is an l-fucose-enriched sulfated polysaccharide isolated from brown algae and marine invertebrates. In this study, we investigated the protective effect of fucoidan from Fucus vesiculosus on alcohol-induced murine liver damage. Liver injury was induced by oral administration of 25% alcohol with or without fucoidan (30 mg/kg or 60 mg/kg) for seven days. Alcohol administration increased serum aspartate aminotransferase and alanine aminotransferase levels, but these increases were suppressed by the treatment of fucoidan. Transforming growth factor beta 1 (TGF-β1), a liver fibrosis-inducing factor, was highly expressed in the alcohol-fed group and human hepatoma HepG2 cell; however, the increase in TGF-β1 expression was reduced following fucoidan administration. Treatment with fucoidan was also found to significantly reduce the production of inflammation-promoting cyclooygenase-2 and nitric oxide, while markedly increasing the expression of the hepatoprotective enzyme, hemeoxygenase-1, on murine liver and HepG2 cells. Taken together, the antifibrotic and anti-inflammatory effects of fucoidan on alcohol-induced liver damage may provide valuable insights into developing new therapeutics or interventions.     

Bacterial colonization and invasion in pigs experimentally exposed to Streptococcus suis serotype 2 in aerosol

A recently developed porcine model for aerogenous infection with Streptococcus suis serotype 2 was applied in a study of the phases of bacterial colonization and initial invasion. Eighteen pigs were exposed to aerosolized S. suis serotype 2 after pre-exposure to mild acetic acid in aerosol. The animals were killed consecutively within the first six days after challenge. After death, all animals were necropsied and examined by bacteriology, histopathology, and immunohistochemistry. Systemic infection was established in four out of 18 animals exposed to S. suis serotype 2. All systemically infected animals developed clinical signs and lesions typical of the infection. In four additional animals, subclinical infection was demonstrated by re-isolation of S. suis from the palatine tonsil. However, in all 18 challenged animals, immunohistochemistry demonstrated S. suis serotype 2 antigen in the palatine and/or nasopharyngeal tonsils. In all four systemically infected animals, S. suis serotype 2 antigen was also found in the mandibular lymph node. These observations point towards the tonsils as possible portals of entry for S. suis serotype 2 with subsequent lymphogenous spread. Thus, the present findings parallel the proposed pathogenesis for S. suis serotype 1 infection in pigs.     

Purification and Characterization of a Hexose Oxidase with Excellent Strengthening Effects in Bread

Hexose oxidase (EC 1.1.3.5) (HOX) was purified 51-fold from the red algae Chondrus crispus, by several chromatography methods, including hydrophobic interaction, chelating Sepharose, anion exchange, gel filtration, and chromatofocusing. Purified HOX was subjected to native PAGE and activity staining with nitroblue tetrazolium. For HOX electroeluted out of the gel and digested with endoproteinase Lys-C, the internal peptide sequence determined was: D-P-G-Y-I-V-I-D-V-N-A-G-T-(V or P)-D-K-P-D-P-X. The molecular mass, determined by gel filtration, was 126 kDa, versus 65 kDa determined by SDS-PAGE. The pI was determined to 4.64 and 4.79 as a double band on an isoelectrofocusing gel. K_m was determined to 2.7 mM for D-glucose, 3.6 mM for D-galactose, 20.2 mM for cellobiose, 43.7 mM for maltose, 90.3 mM for lactose, 102 mM for xylose, and 531 mM for arabinose. The oxidation of thiol groups in gluten was determined by using Ellman's reagent: 5,5′-dithiobis (2-nitrobenzoic acid). The effect of HOX was compared to that of glucose oxidase. Both enzymes caused a dose-responsive reduction in the free thiol groups. Extensigraph measurements and baking tests confirmed that HOX caused increased dough strength and increased bread volume more efficiently than glucose oxidase used in the same dosage.     

Seasonal variation of provitamin D2 and vitamin D2 in perennial ryegrass (Lolium perenne L.)

Ergosterol (provitamin D(2)) is converted to vitamin D(2) in grass by exposure to UV light. Six varieties of perennial ryegrass (Lolium perenne L.) were harvested four times during the season, and the contents of vitamin D(2) and ergosterol were analyzed by a sensitive and selective liquid chromatography tandem mass spectrometry method. Weather factors were recorded, and a principal component analysis was performed to study which factors were important for the formation of vitamin D(2). The results suggest that a combination of weather factors is involved and that the contents of ergosterol and vitamin D(2) change more than a factor of 10 during the season. These results demonstrate that grass potentially can be a significant source of vitamin D for grazing animals and animals fed on silage and hay.     

Quality control using a multilevel logistic model for the Danish pig Salmonella surveillance antibody-ELISA programme

In Denmark, the level of Salmonella infection in pig herds is monitored with a surveillance programme using an indirect antibody ELISA. Our purpose with the present study was to determine whether sample results from the programme were useful in the quality control of this ELISA. Test results from the year 2003, in which the laboratory experienced a technical problem with an automatic microtitre-plate washing machine, were examined statistically. We chose 3 months for the analysis: January, where the problem was moderate, June with the problem more serious, and November, where the problem had been solved. A logistic analysis was carried out with outcome 0 for a negative test result and 1 for a positive test result. Row and column on the microtitre plates, multiprobe robot, and their interactions were included as fixed effects, and date, plate, and slaughterhouse were included as random effects. Backward elimination was carried out using alpha=0.05 to achieve a final model for each month. The row and the column were significant in January and June, and a robot effect was also included in the model for January. In June, an interaction between row and column was identified. In November, none of the fixed effects was significant. Breaking the months January and June into shorter time intervals showed that the row and column effects were significant also when data were from only 1 week, whereas the robot main effect was not significant in most periods and the interaction effects were not significant throughout. Analysis of the test results from the wells with test samples gave good information on systematic errors across the microtitre plates, and severe errors appeared significant even when data from short time periods were used.     

The Importance of Nitrogen Oxides for the Exceedance of Critical Thresholds in the Nordic Countries

Impacts of air pollutants and especially acidification in ecosystems have been of serious concern in the Nordic countries since the 1970s. The current approach to assess several pollutants (sulfur and nitrogen oxides, ammonia, volatile organic compounds) and their effects (acidification, eutrophication and ground-level ozone) simultaneously is extremely complex. This study explored the relative role of nitrogen oxides in environmental impacts in the Nordic countries. The share of NO_x in the exceedances of critical loads, the long-term ecosystem protection targets, was found to be roughly 25% in acidification and 50% in eutrophication. The contribution of NO_x emissions to ground-level ozone formation was considered important, as NO_x is the limiting precursor in ozone formation in the Nordic countries. The comparison of observed and modeled accumulated ozone concentrations (AOT40) for the early 1990s shows noticeable differences in the Nordic area, partly due to the sensitivity of the AOT40 indicator to the 40 ppb threshold value.