The influence of salmon surface mucus on the growth of Flavobacterium columnare

Flavobacterium columnare is the causative agent of columnaris disease. The presence of lesions on the gills, skin and fins of diseased fish suggests that F. columnare is able to utilize fish skin mucus as a substrate for growth and that exposure to this material would alter the expression of genes involved in the colonization of the outer surfaces of the fish. Growth, biofilm formation, extracellular protease production and changes in protein expression of F. columnare strain C#2 cultured in media supplemented with juvenile Atlantic salmon skin mucus were compared with the same media without mucus. C#2 was able to grow by using mucus as the sole nutrient source. Growth in mucus-containing media induced cells to grow as a biofilm and extracellular protease activity increased in mucus-containing cultures. SDS-PAGE protein profiles showed that expression of six extracellular proteins increased in mucus-containing media. These results demonstrate that salmon surface mucus promotes the growth of F. columnare and that exposure to mucus alters the growth characteristics of this bacterium with regard to protease production and biofilm formation. Further characterization of mucus-induced physiological changes will increase our understanding of the basis of virulence of this economically important fish pathogen.     

The solubilization of myofibrillar proteins of vertebrate skeletal muscle in water

Myofibrillar proteins of vertebrate skeletal muscles are insoluble in solutions of ionic strength that approximate physiological conditions. We established a method to solubilize more than 80% of chicken breast muscle myofibrillar proteins in water for the use of meat as a source of food protein. SDS‐polyacrylamide gel electrophoretic patterns of water‐soluble myofibrillar proteins demonstrated that all identified myofibrillar proteins except connectin/titin were soluble in water. A part of α‐actinin was released from myofibrils by repeated washing with 2.5 mmol/L NaCl and 5 mmol/L L‐histidine solution, and subsequent destruction of connectin/titin in washed myofibrils by ultrasonication resulted in solubilization of a large fraction of chicken breast muscle myofibrillar proteins in water. Myofibrillar proteins of chicken leg, pork loin, beef shoulder loin, and lamb were also solubilized in water using this procedure.     

Is the nutritive value of tall fescue (Festuca arundinacea Schreb.) related to the accumulated forage mass?

The aim of this work was to investigate whether neutral detergent fibre (NDF) and dry‐matter digestibility (DMD) are related to tall fescue accumulated forage mass (AFM) and to assess the relevance of environmental variables to predict the nutritive value of tall fescue swards. Three experiments were carried out in Pergamino, Argentina. To obtain swards with different amounts of AFM, two N levels and two irrigation regimes were applied in the spring after sowing and the autumn of the next year. In spring and autumn, AFM, NDF and DMD were measured every 10–12 days. In spring, NDF increased from 503 to 604 g kg^?1, DMD decreased from 684 to 558 g kg^?1 and AFM increased from 0·64 to 2·82 t DM ha^?1. In autumn, NDF decreased from 543 to 442 g kg^?1, DMD increased from 591 to 681 g kg^?1 and AFM increased from 0·35 to 1·10 t DM ha^?1. The results show that the nutritive value of tall fescue through the year is not related to the accumulation of dry matter of the sward. Nutritive value is determined by the reproductive stage in late spring and early summer, the fate of photosynthates at different times of the year and the synthesis of non‐digestible compounds.     

蛹虫草CmKexin基因克隆和菌丝体中表达检测

为了探讨蛹虫草类枯草杆菌蛋白酶(Subtilisin-like protease)的表达特性,以真菌蛹虫草菌丝体为研究对象,通过RT-PCR获得蛹虫草CmKexin基因的ORF序列,并进行序列分析。应用Northern杂交和Real-time PCR方法,检测蓝光照射后蛹虫草菌丝体中CmKexin基因的转录情况。结果获得蛹虫草CmKexin基因的ORF序列。对翻译蛋白质产物CmKexin进行生物信息学分析,表明该蛋白质含1个属蛋白转化酶的肽酶S8家族功能域(143~429)和1个前体蛋白转化酶P功能域(514~600),符合真菌类枯草杆菌蛋白酶的特征。蛹虫草菌丝体在黑暗预培养4 d后再蓝光照射,Northern Blotting和Real-time PCR检测均可得到相同的结果,即在持续的蓝光照射48~50 h时,出现CmKexin基因的瞬时大量转录。而其他时间内均未检测到该基因的大量转录。试验结果将为蛹虫草类枯草杆菌蛋白酶的利用提供理论依据。     

Intracellular and extracellular enzyme activity in soil with reference to elemental cycling

Enzyme activities play an important role for the transformation of elements and compounds in soil and, thus, were extensively analyzed for more than 4 decades. The activity of any enzyme in soil may not only be controlled by active organisms. Substantial parts of ‘extracellular’ enzymes may be stabilized by abiotic soil components maintaining their activity. Methods to discriminate the source of enzyme activity were summarized with emphasis on the approach plotting enzyme activity versus a feature integrating the microbial biomass after the addition of glucose and nitrate. Considering the quotient between enzyme activity and microbial biomass content, protease activity will be discussed with reference to nitrogen transformation in soils.     

汽车发电机电动势3次谐波分量的估算

分析了Y联接和Y0联接整流电路在畸变电动势作用时的中性点电压,给出了畸变电动势3次谐波分量的估算方法,并估算了Y0联接汽车交流发电机在中性点二极管导通时电动势中3次谐波分量的大小.     

Preparation and antinutritional characteristics of dry bean (Phaseolus vulgaris L.) protein concentrates

Protein concentrates and starches were prepared by a wet extraction process from five dry bean (Phaseolus vulgaris L.) cultivars. The protein contents ranged from 69.7–76.4%. Concentrates prepared from dehulled beans under similar conditions had higher protein contents (80.6–87.9%). Each additional washing of the concentrates with distilled water increased their protein content. However, the protein recovery progressively decreased. The yield of starch ranged from 48.0–51.1% of the starting material. The solubility of bean proteins was minimal at pH 4.0, and under alkaline conditions, it was influenced by the tannin contents of the concentrates. Protein concentrates had lower trypsin, chymotrypsin, and amylase inhibitory activities as well as lower phytic acid and tannin contents compared to whole bean flours.     

Efficient Screening of Marine Extracts for Protease Inhibitors by Combining FRET Based Activity Assays and Surface Plasmon Resonance Spectroscopy Based Binding Assays

The screening of extracts from marine organisms is a widely used strategy to discover new drug leads. A common problem in the screening process is the generation of false positive hits through unspecific effects from the complex chemical composition of the crude extracts. In this study, we explored a combination of a fluorescence resonance energy transfer (FRET) based activity assay and a surface plasmon resonance (SPR) based binding assay to avoid this problem. An aqueous extract was prepared from rest raw material of the Norwegian spring spawning herring, and further fractionated by methanol solubility and solid phase extraction. FRET based activity assays were used to determine the influence of each extract on the activity of different proteases. Several extracts showed more than 50% inhibition. The inhibition mechanisms were elucidated by SPR based competition experiments with known inhibitors. For the secreted aspartic proteases 1, 2, 3 and HIV-1 protease, the results indicated that some extracts contain inhibitors interacting specifically with the active site of the enzymes. The study shows that a combination of an activity assay and an SPR based binding assay is a powerful tool to identify potent inhibitors in marine extracts. Furthermore, the study shows that marine vertebrates offer an interesting source for new bioactive compounds, although they have rarely been explored for this purpose.