Attenuation of allergic contact dermatitis through the endocannabinoid system

Allergic contact dermatitis affects about 5% of men and 11% of women in industrialized countries and is one of the leading causes for occupational diseases. In an animal model for cutaneous contact hypersensitivity, we show that mice lacking both known cannabinoid receptors display exacerbated allergic inflammation. In contrast, fatty acid amide hydrolase-deficient mice, which have increased levels of the endocannabinoid anandamide, displayed reduced allergic responses in the skin. Cannabinoid receptor antagonists exacerbated allergic inflammation, whereas receptor agonists attenuated inflammation. These results demonstrate a protective role of the endocannabinoid system in contact allergy in the skin and suggest a target for therapeutic intervention.     

GOAnnotator: linking protein GO annotations to evidence text

Background

Annotation of proteins with gene ontology (GO) terms is ongoing work and a complex task. Manual GO annotation is precise and precious, but it is time-consuming. Therefore, instead of curated annotations most of the proteins come with uncurated annotations, which have been generated automatically. Text-mining systems that use literature for automatic annotation have been proposed but they do not satisfy the high quality expectations of curators.

Results

In this paper we describe an approach that links uncurated annotations to text extracted from literature. The selection of the text is based on the similarity of the text to the term from the uncurated annotation. Besides substantiating the uncurated annotations, the extracted texts also lead to novel annotations. In addition, the approach uses the GO hierarchy to achieve high precision. Our approach is integrated into GOAnnotator, a tool that assists the curation process for GO annotation of UniProt proteins.

Conclusion

The GO curators assessed GOAnnotator with a set of 66 distinct UniProt/SwissProt proteins with uncurated annotations. GOAnnotator provided correct evidence text at 93% precision. This high precision results from using the GO hierarchy to only select GO terms similar to GO terms from uncurated annotations in GOA. Our approach is the first one to achieve high precision, which is crucial for the efficient support of GO curators. GOAnnotator was implemented as a web tool that is freely available at http://xldb.di.fc.ul.pt/rebil/tools/goa/.

     

Fermentation enhances the biological activity of Allium cepa bulb extracts

In this study, we compared the analytical fingerprint and bioactivity of three onion extracts, including an aqueous, a methanol, and a fermented aqueous extract. The extracts were characterized by HPLC-DAD, LC-MS, and GC-MS analyses. The antibacterial, antigenotoxic, and antiproliferative activity of these extracts was assessed by means of agar disk diffusion, bacterial growth kinetics, a comet assay, cell cycle distribution analysis, and cell viability testing. Both the aqueous and methanolic extracts showed a typical flavonol-fingerprint as assessed by HPLC measurements and showed little to no bioactivity. The fermented aqueous extract, which lacks the usual onion flavonoid profile, was found to be the most active in all of the assays. This finding indicates that metabolites of onion compounds, generated by lactic acid fermentation, may be more active than their precursor substances.     

Physicochemical properties of native and recombinant mungbean (Vigna radiata L. Wilczek) 8S globulins and the effects of the N-linked glycans

We have previously cloned and characterized the cDNAs of three isoforms of the 8S globulin of mungbean, expressed the major 8Salpha isoform in Escherichia coli, and purified and successfully crystallized it (Bernardo, A. E. N.; Garcia, R. N.; Adachi, M.; Angeles, J. G. C.; Kaga, A; Ishimoto, M.; Utsumi, S.; Tecson-Mendoza, E. M. J. Agric. Food Chem. 2004, 52, 2552-2560). Herein, we report the physicochemical and emulsifying properties of the native 8S and recombinant 8Salpha globulin or vicilin. The circular dichroism spectra analysis of the native 8S and recombinant 8Salpha globulins revealed that the recombinant 8Salpha formed a secondary structure close to that of the native 8S. Further, gel filtration analysis showed that 8Salpha was able to assemble into trimers. The native 8S and recombinant 8Salpha globulins were soluble at pH 3.4 and at pH 7.4-9.0 at low ionic strength, mu = 0.08. Interestingly, the native 8S was more soluble at pH 7.0 and pH 7.4 than the recombinant 8Salpha at mu = 0.08. Both forms were very soluble at pH 3.4-9.0 at high ionic strength, mu = 0.50. The native form exhibited a higher T(m) (69.2, 79.5, and 83.8 degrees C) than the recombinant form (65.6, 71.6, 77.5 degrees C) at mu = 0.1, 0.2, and 0.5, respectively. The recombinant form was found to have greater surface hydrophobicity than the native form. There was little difference in the emulsifying ability between the native 8S and 8Salpha at pH 3.4 and pH 7.6. The results indicate that the presence of N-linked glycans is not essential in the assembly and stable conformation of the mungbean vicilin. However, the N-linked glycans might have contributed to the higher solubility at low ionic strength, greater thermal stability, and decreased surface hydrophobicity of the native vicilin as compared to the recombinant 8Salpha. On the other hand, the N-linked glycans showed little effect on the emulsifying ability of the protein.     

Caprine hepatic lipidosis induced through the intake of low levels of dietary cobalt

Forty-one, 10-week-old newly weaned goats were randomly allocated into two groups, namely control (n=22) and treated (n=19). Kids in both groups were fed Rhodegrass hay ad libitum that contained < 0.1 mg/kg DM cobalt and 150 g/day of a commercially prepared ruminant concentrate that contained approximately 0.12 mg/kg DM cobalt. This diet provided the minimum daily requirement of cobalt as specified for sheep. The treated goats were supplemented with bi-monthly subcutaneous injections of 2000 microg of hydroxycobalamin. All goats were weighed and blood samples collected monthly for haematological, clinical biochemical and serum vitamin B12 analysis. After a 10-month experimental period the goats were slaughtered. The control animals exhibited significantly (P<0.05) lower weight gains, and had dry scruffy hair coats. In addition, there was a decline in erythrocyte counts, mean haemoglobin, packed cell volume, mean corpuscular volume, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration. Controls also exhibited significantly (P<0.05) lower levels of total serum proteins and elevated levels of serum alkaline phosphatase compared to treated goats. Fourteen (63.6%) of the control goats developed pathology consistent with reported field cases of hepatic lipidosis associated with low liver levels of cobalt. Only one (5.3%) of the treated goats developed hepatic lipidosis. Contrary to previous reports that suggested that goats are less sensitive to low levels of dietary cobalt than sheep, it is apparent that this is not the case with Omani goats. This is the first report of the induction of hepatic lipidosis in goats due to feeding low levels of cobalt in their diet.     

Enhancing resistance of transgenic carrot to fungal pathogens by the expression of Pseudomonas fluorescence microbial factor 3 (MF3) gene

Microbial factor 3 (MF3) gene from a plant-growth promoting rhizobacteria Pseudomonas fluorescence was introduced into carrot genome using Agrobacterium-mediated transformation. The obtained transgenic plants expressing MF3 protein were grown in a glasshouse and evaluated for their resistance to phytopathogens. The assays were performed by the assessment of disease symptoms developing on the detached leaflets and petioles inoculated by three fungal pathogens. The results showed that transgenic plants had significantly enhanced resistance to Alternaria dauci, Alternaria radicina and Botrytis cinerea, on average, by 20–40% in comparison to the non-transformed control plants. This is the first report of enhancing plant resistance by expressing the bacterial protein homologous to the family of FK506-binding proteins.     

Soil CO2 evolution rates in the field – a comparison of three methods

In this field study, three methods for determining soil CO₂ evolution rates were compared: (1) a static chamber method and (2) a dynamic chamber method, both with gas chromatographic (GC) analysis of air samples, and (3) a dynamic chamber system using a portable infrared gas analyzer (CIRAS). The mean CO₂ evolution rate in the field increased in the order static chamber-GC < dynamic chamber-GC < dynamic chamber-CIRAS by 40%. The CO₂ evolution rates obtained using the three methods were significantly correlated. None of the three methods was significantly affected by soil moisture or soil temperature. This has been repeatedly observed by others, but is still not fully understood. The difference between air and soil temperature had strong negative effects on both GC methods, but not on the CIRAS system.