Identification and molecular characterization of two tandemly located flagellin genes from Aeromonas salmonicida A449

Two tandemly located flagellin genes, flaA and flaB, with 79% nucleotide sequence identity were identified in Aeromonas salmonicida A449. The fla genes are conserved in typical and atypical strains of A. salmonicida, and they display significant divergence at the nucleotide level from the fla genes of the motile species Aeromonas hydrophila and Aeromonas veronii biotype sobria. flaA and flaB encode unprocessed flagellins with predicted Mrs of 32,351 and 32,056, respectively. When cloned under the control of the Ptac promoter, flaB was highly expressed when induced in Escherichia coli DH5alpha, and the FlaB protein was detectable even in the uninduced state. In flaA clones containing intact upstream sequence, FlaA was barely detectable when uninduced and poorly expressed on induction. The A. salmonicida flagellins are antigenically cross-reactive with the A. hydrophila TF7 flagellin(s) and evolutionarily closely related to the flagellins of Pseudomonas aeruginosa and Vibrio anguillarum. Electron microscopy showed that A. salmonicida A449 expresses unsheathed polar flagella at an extremely low frequency under normal laboratory growth conditions, suggesting the presence of a full complement of genes whose products are required to make flagella; e.g., immediately downstream of flaA and flaB are open reading frames encoding FlaG and FlaH homologs.     

一种新型的无线网络入侵检测方法-动态自适应模板法

为了能够检测到无线网络系统中的已知和未知类型的入侵者,提高无线网络系统的安全性,本文提出了动态自适应模板法。该方法的基本思想是原有模板(分类结果)在聚类过程中不断更新,并且允许在聚类分析过程中构成新的模板。实验表明用该方法检测无线网络系统中的新型入侵者,准确率可达到98%。     

Measurement of anthocyanins and other phytochemicals in purple wheat

The major anthocyanin composition of normal purple wheat and heat stressed purple wheat were measured using HPLC, LC–MS/MS and the pH differential method. The lignan secoisolariciresinol diglucoside (SDG) and melatonin content were also measured. Total anthocyanin profile of normal purple wheat (491.3 mg/kg) was significantly (P < 0.05) lower than that of the heat stressed purple wheat (522.7 mg/kg). Thirteen major anthocyanins were isolated and cyanidin 3-glucoside was the predominant anthocyanin in purple wheat. Using the pH differential method, the total anthocyanin content of normal (500.6 mg/kg) and heat stressed (526.0 mg/kg) purple wheat were similar to those observed using HPLC. The SDG content of normal and heat stressed purple wheat were 770 and 520 μg/kg, while melatonin content was 4 and 2 μg/kg, respectively. The presence of SDG and melatonin in addition to anthocyanins may contribute to the health benefits associated with consumption of coloured cereal grains.     

Antioxidant properties of commercial, regular- and whole-wheat spaghetti

Whole grains contain more vitamins, minerals, natural antioxidants and dietary fibre than regular, refined grain products. Therefore, consumption of whole grain products is associated with beneficial health effects. The present investigation evaluated the antioxidant properties of 10 samples of regular- and whole-wheat spaghetti that are commercially available. The methods employed were total phenolic content (TPC), 2,2-diphenyl-1-picryhydrazyl (DPPH) free radical scavenging activity, oxygen radical absorbance capacity (ORAC) and ferulic acid content by HPLC analysis. The effects of cooking on the antioxidant properties of spaghetti were also studied. Whole wheat spaghetti exhibited significantly higher levels of total phenolic content (1389 μg/g) than regular wheat spaghetti (865 μg/g); however, TPC in both regular and whole wheat spaghetti was 48–78% of the original content after cooking. There were no significant differences in ORAC values (34.3–100.4 μmol Trolox equivalents/g) or DPPH scavenging activity (1.0–2.3 μmol Trolox equivalents) among whole wheat and regular spaghetti. Whole wheat spaghetti (234 μg/g) had significantly higher content of ferulic acid than regular spaghetti (p < 0.05). TPC and ferulic acid content were found to be good indicators of the antioxidant capacity of spaghetti with both indices demonstrating the superiority of whole wheat over regular pasta products. The current findings on spaghetti add to the mounting evidence on the potential health benefits to be derived from consuming whole grain products.     

Effect of pre-dehulling treatments on some nutritional and physical properties of navy and pinto beans (Phaseolus vulgaris L.)

The effect of pre-dehulling treatments using low and high temperatures on some nutritional and physical properties of navy and pinto beans was investigated. Beans were exposed to water (14%, 28% and soaking 1:5, w/v for 6 h, 16 and 16 h, respectively) to facilitate seed coat detachment prior to freeze drying (FD) for 48 h or heat drying (HT) for 20 or 60 min. Exposure to the highest moisture levels produced the largest seed coat yields (17.38-20.91%) and was independent of the drying conditions. The total phenolic content was positively correlated to the 1,1-diphenyl-2-picrylhydrazyl (DPPH) antioxidant activity and increased as the exposure time to HT increased, but it was unaffected by the FD. The nutritional properties of the bean varieties differed significantly, phytic acid was unaffected, and the highest antioxidant activity was observed on the seed coats of HT pinto beans (69.24-84.46% of DPPH discoloration). Significant physical changes were observed for the HT seeds, with the highest yellowness and the lowest peak viscosity detected in the soaked and HT (60 min) beans.     

ESI-MS compatible permanent coating of glass surfaces using poly(ethylene glycol)-terminated alkoxysilanes for capillary zone electrophoretic protein separations

Thin poly(ethylene glycol) silane (PEG-silane) coatings formed from N-(triethoxysilyl propyl)-O-poly(ethylene oxide) urethane with different chain lengths of poly(ethylene glycol) (MW 750 and 4000-5000) are used to modify glass microfluidic channels and fused-silica capillaries for electrophoretic separations of proteins. These coatings combine three important properties, which make them favorable for proteomic analyses including reduction of protein adsorption, compatibility with mass spectrometry due to their stability, and the ability to control the magnitude of electroosmotic flow (EOF). The coatings have been successfully used in microfluidic chips and fused-silica capillaries for separation of protein sample mixtures under low EOF conditions. The long-chain and mixed PEG-silane coatings suppress electroosmotic flow by more than 90%, whereas the short-chain PEG silane suppresses EOF by 65-75% at pH values of 3-9. The long-chain and mixed PEG-silane coatings are suitable for low EOF applications or for cases where negative effects of EOF are to be minimized. Efficient separations of unlabeled basic proteins at low pH and FITC-labeled proteins at high pH were achieved, as well as excellent stability for at least 200 electrophoretic runs. Additionally, these covalent coatings produce no detectable background ions in ESI-MS, making them compatible with on-line mass spectrometry.     

An evaluation of carotenoid levels and composition of glabrous canaryseed

Nineteen glabrous canaryseeds comprising brown- and yellow-coloured varieties were investigated to determine carotenoid content and their properties. Total carotenoid content of canaryseeds ranged from 7.57 to 11.69 mg/kg. β-Carotene, lutein, and zeaxanthin were the major carotenoids of glabrous canaryseed. β-Carotene ranged from 5291 to 6273, 4564 to 5352, and 3651 to 4428 μg/kg while lutein ranged from 2667 to 3370, 1532 to 3007, and 2042 to 2299 μg/kg, respectively in canaryseed flour, wholemeal and bran. Zeaxanthin was relatively low (?650 μg/kg). High β-carotene levels distinguished glabrous canaryseeds from other cereal crops as potential ingredients for carotenoid-enriched functional foods.     

Qualitative and quantitative analysis of the major phenolic compounds as antioxidants in barley and flaxseed hulls using HPLC/MS/MS

BACKGROUND: Both qualitative and quantitative analyses of the major phenolic compounds in barley and flaxseed hulls were conducted using reverse phase high‐performance liquid chromatography coupled with photodiode array detection and quadrupole time‐of‐flight mass spectrometry. RESULTS: Ferulic acid, p‐coumaric acid, vanillic acid and vanillin were identified and quantified in four barley hull samples. Four ferulate dehydrodimers were also detected. The phenolic compounds of flaxseed hull were distinct from those of barley hull. Three flaxseed hull samples varied significantly (P < 0.05) in their contents of secoisolariciresinol diglucoside (16.38–33.92 g kg^?1), coumaric acid glucoside (35.68–49.22 g kg^?1) and ferulic acid glucoside (5.07–15.23 g kg^?1). The phytochemical profiles of co‐extracts featured the major phenolic compounds from both barley and flaxseed hulls. The total phenolic content and 2,2‐diphenyl‐1‐picrylhydrazyl radical‐scavenging capacity varied significantly (P < 0.05) among different varieties of flaxseed and barley hulls. CONCLUSION: As agricultural by‐products, barley and flaxseed hulls may be utilised as potential sources of functional food ingredients through extraction and concentration of the phytochemicals identified above. Copyright © 2012 Society of Chemical Industry     

Effect of chemical conditioning on the milling of high‐tannin sorghum

Suitable methods are required for milling tannin‐containing sorghums. Sorghum varieties SV2 (tannin‐free), Chirimaugute (medium‐tannin) and DC‐75 (high‐tannin) were milled using a simple roller mill and a multi‐sample tangential abrasive decortication device. Grain was conditioned up to 20% moisture prior to milling using HCl (0.9%, v/v), formaldehyde (HCHO; 0.05%, v/v), NaOH (0.3%, w/v) and water as control. Abrasive decortication and roller milling reduced levels of the polyphenols. Polyphenol reduction was affected significantly by variety, chemical treatment and conditioning moisture (P < 0.001). NaOH and HCHO treatments gave lower polyphenol content in the meal and offal after roller milling. Enzyme inhibition (EI) by polyphenols was reduced by 52% after decortication or roller milling. Chemical treatment did not significantly affect EI levels after decortication. NaOH and HCHO treatments gave the lowest EI when Chirimaugute was roller milled at 20% moisture. The yield of product obtained after decortication was high at 12% moisture for SV2 and Chirimaugute and at 16% conditioning moisture for DC‐75. Conditioning to 20% moisture prior to milling did not improve product yield. Roller milling and decortication resulted in products that were lighter in colour than the grain. Both abrasive decortication and roller milling plus NaOH and HCHO reduced tannin content of offal. It is concluded that conditioning using NaOH could be advantageous in roller milling but not in abrasive decortication of tannin‐containing sorghums. © 2000 Society of Chemical Industry     

Effect of thermal processing on antioxidant properties of purple wheat bran

Antioxidant activity of purple wheat bran, heat-treated purple wheat bran, and purple wheat bran muffins was evaluated to determine the impact of thermal processing on potential health benefits. The purple wheat bran and muffin samples were analyzed for total phenolic content, anthocyanin content and free radical scavenging activity using peroxyl (oxygen radical absorbance capacity assay) and 2,2-diphenyl-1-picrylhydrazyl (DPPH assay) radicals. Total phenolic content and oxygen radical absorbance capacity (ORAC) values of sample extracts were significantly affected by various extracting solvents. The conditions selected for heat treatment did not markedly change antioxidant activity of purple wheat bran. However, there was a significant reduction in total phenolic contents, ORAC values and total anthocyanins during processing of purple wheat bran- or heat-treated purple wheat bran-enriched muffins. On the contrary, muffin extracts still remained excellent in DPPH radical scavenging activity.