Production and characterization of neutralizing monoclonal antibodies that recognize an epitope in domain 2 of Bacillus anthracis protective antigen

Antibodies against the protective antigen (PA) of Bacillus anthracis play a key role in response to infection by this important pathogen. The aim of this study was to produce and characterize monoclonal antibodies (mAbs) specific for PA and to identify novel neutralizing epitopes. Three murine mAbs with high specificity and nanomolar affinity for B. anthracis recombinant protective antigen (rPA) were produced and characterized. Western immunoblot analysis, coupled with epitope mapping using overlapping synthetic peptides, revealed that these mAbs recognize a linear epitope within domain 2 of rPA. Neutralization assays demonstrate that these mAbs effectively neutralize lethal toxin in vitro.     

Stochastic dynamics of magnetosomes and a mechanism of biological orientation in the geomagnetic field

The rotations of nanoscopic magnetic particles, magnetosomes, embedded into the cytoskeleton are considered. Under the influence of thermal disturbances, a great number of magnetosomes are shown to move chaotically between two stable equilibrium positions, in which their magnetic moments are neither parallel nor antiparallel to the static Earth's magnetic field (MF). The random rotations attain the value of order of a radian. The rate of the transitions and the probability of magnetosomes to be in the different states depend on the MF direction with respect to an averaged magnetosome's orientation. This effect explains the ability of migratory animals to orient themselves faultlessly in long term passages in the absence of the direct visibility of optical reference points. The sensitivity to deviation from an "ideal" orientation is estimated to be 2-4 degrees. Possible involvement of the stochastic dynamics of magnetosomes in biological magnetic navigation is discussed.     

Cry1Ac杀虫蛋白对粘虫中肠几种酶活性的影响

为阐明Bt杀虫蛋白对次要靶标害虫粘虫Mythimna separata (Walker) （鳞翅目： 夜蛾科）的生理学影响, 本研究分析比较了粘虫高龄幼虫在室内取食低剂量Cry1Ac杀虫蛋白6, 12, 24和36 h后, 其体内主要的解毒酶（酯酶和谷胱甘肽-S-转移酶）、 保护酶（超氧化物歧化酶、 过氧化氢酶和过氧化物酶）和中肠蛋白酶（总蛋白酶、 强碱性类胰蛋白酶、 弱碱性类胰蛋白酶和类胰凝乳蛋白酶）等活性的变化。结果表明, 取食Cry1Ac杀虫蛋白后, 粘虫幼虫体内相关酶活力呈现不同的变化趋势： （1）酯酶、 谷胱甘肽-S-转移酶、 过氧化物酶(POD)、 类胰蛋白酶和类胰凝乳蛋白酶活力较对照显著降低（P<0.05）; （2）超氧化物歧化酶(SOD) 活力较对照显著升高（P<0.05）; （3）过氧化氢酶(CAT) 活力于6, 12和24 h显著低于对照（P<0.05）, 36 h时显著高于对照（P<0.05）。结果提示Cry1Ac杀虫蛋白主要通过抑制粘虫幼虫中肠解毒酶和蛋白酶的活性, 扰乱SOD, CAT 和POD 3种保护酶的动态平衡而干扰幼虫的正常生理代谢, 从而起到毒杀粘虫的作用。     

RIG‐I detects infection with live Listeria by sensing secreted bacterial nucleic acids

Immunity against infection with Listeria monocytogenes is not achieved from innate immune stimulation by contact with killed but requires viable Listeria gaining access to the cytosol of infected cells. It has remained ill‐defined how such immune sensing of live Listeria occurs. Here, we report that efficient cytosolic immune sensing requires access of nucleic acids derived from live Listeria to the cytoplasm of infected cells. We found that Listeria released nucleic acids and that such secreted bacterial RNA/DNA was recognized by the cytosolic sensors RIG‐I, MDA5 and STING thereby triggering interferon β production. Secreted Listeria nucleic acids also caused RIG‐I‐dependent IL‐1β‐production and inflammasome activation. The signalling molecule CARD9 contributed to IL‐1β production in response to secreted nucleic acids. In conclusion, cytosolic recognition of secreted bacterial nucleic acids by RIG‐I provides a mechanistic explanation for efficient induction of immunity by live bacteria.     

In utero and early-life exposure of rats to a Wi-Fi signal: screening of immune markers in sera and gestational outcome

An experimental approach was used to assess immunological biomarkers in the sera of young rats exposed in utero and postnatal to non-ionizing radiofrequency fields. Pregnant rats were exposed free-running, 2 h/day and 5 days/week to a 2.45 GHz Wi-Fi signal in a reverberation chamber at whole-body specific absorption rates (SAR) of 0, 0.08, 0.4, and 4 W/kg (with 10, 10, 12, and 9 rats, respectively), while cage control rats were kept in the animal facility (11 rats). Dams were exposed from days 6 to 21 of gestation and then three newborns per litter were further exposed from birth to day 35 postnatal. On day 35 after birth, all pups were sacrificed and sera collected. The screening of sera for antibodies directed against 15 different antigens related to damage and/or pathological markers was conducted using enzyme-linked immunosorbent assay (ELISA). No change in humoral response of young pups was observed, regardless of the types of biomarker and SAR levels. This study also provided some data on gestational outcome following in utero exposure to Wi-Fi signals. Mass evaluation of dams and pups and the number of pups per litter was monitored, and the genital tracts of young rats were observed for abnormalities by measuring anogenital distance. Under these experimental conditions, our observations suggest a lack of adverse effects of Wi-Fi exposure on delivery and general condition of the animals.     

Farm animal milk proteomics

Milk is one of the most important nutrients for humans during lifetime. Farm animal milk in all its products like cheese and other fermentation and transformation products is a widespread nutrient for the entire life of humans. Proteins are key molecules of the milk functional component repertoire and their investigation represents a major challenge. Proteins in milk, such as caseins, contribute to the formation of micelles that are different from species to species in dimension and casein-type composition; they are an integral part of the MFGM (Milk Fat Globule Membrane) that has being exhaustively studied in recent years. Milk proteins can act as enzymes or have an antimicrobial activity; they could act as hormones and, last but not least, they have a latent physiological activity encoded in their primary structure that turns active when the protein is cleaved by fermentation or digestion processes. In this review we report the last progress in proteomics, peptidomics and bioinformatics. These new approaches allow us to better characterize the milk proteome of farm animal species, to highlight specific PTMs, the peptidomic profile and even to predict the potential nutraceutical properties of the analyzed proteins.     

活性炭对苦荞幼苗根系和叶片生理特性的影响

以山西省主栽苦荞品种‘黑丰1号’温室盆栽幼苗为材料,设置土壤活性炭含量分别为0(CK)、2.5(B2.5)、5.0(B5.0)、7.5(B7.5)、10(B10)g/kg共5个水平,研究土壤中施加活性炭后对苦荞幼苗根系及碳氮代谢、保护酶活性等指标的影响.结果显示:(1)随着活性炭施用比例的增加,苦荞幼苗根系生长指标和根系活力指标均呈先增后减的趋势,根平均直径呈先减后增的趋势,其中B5.0、B7.5处理的幼苗根系总长度、总表面积、总体积、活跃吸收面积、根尖数均显著高于对照,但B10处理的根系发育减弱.(2)随活性炭施用比例的增加,苦荞幼苗叶片蔗糖酶活性变化呈先增后减的趋势,同一处理水平条件下随苦荞的生长而逐渐下降;B2.5、B5.0处理苦荞幼苗叶片蔗糖酶活性和可溶性糖含量均比CK极显著增加,B7.5处理略有提高,B10处理差异不显著.(3)苦荞幼苗叶片谷氨酰胺合成酶(GS)活性随活性炭的增加基本呈上升趋势,而同一处理水平下随苦荞的生长而下降;叶片GS活性在B5.0、B7.5处理时比CK极显著提高,可溶性蛋白质含量在B7.5处理时也显著提高.(4)叶片保护酶SOD、POD、CAT活性随活性炭浓度的升高呈先升后降的变化趋势,而同一处理水平下各时期间变化不大;B2.5处理叶片的SOD、POD和CAT活性比对照显著增强.研究发现,适量施用活性炭(2.5～7.5g/kg)能有效促进苦荞幼苗碳氮代谢和保护酶活性,增强其根系活力.     

添加保护剂促进碱性果胶酶的工业保藏稳定性

保藏稳定性对于商品酶具有重要的实际意义。通过对碱性果胶酶的保护剂进行研究,最终筛选得到最优复合保护剂配方为：Tween201mL／L,六偏磷酸钠1．5g／L,明胶5g／L,甘油5％（口～）,CaCl22g／L;热稳定性提高18倍。此外在常温保藏时,在酶液中添加0．3‰山梨酸钾和0．3‰苯甲酸钠,能有效提高保藏效果,减少酶液染茵、混浊和发臭,最终常温保藏90d后酶活保留率为84．5％,达到工业化保藏的要求。