Cross-reactivity between peanut and lupin proteins

Peanut-allergic individuals may also react to lupin, which, for this reason, has been included in the EU list of food allergens. Since there is not yet any general consensus on the major allergen/s in lupin, the objective of this investigation was to compare the reactivity of the main lupin proteins towards the IgE of the sera of allergic patients. Both Lupinus albus and Lupinus angustifolius were investigated. ELISA’s, Western blotting and mass spectrometry, including also de novo sequencing of the unknown lupin proteins, were used for identifying the IgE-binding proteins. Significant differences in the protein reactivities were observed. In particular, there was a direct relationship between the level of peanut-specific IgE’s and the cross-reactivity to lupin proteins; also the reactivity of each serum appeared to be unique. Although numerous lupin proteins bind IgE’s, our data suggest a predominant contribution of α-conglutin in the reactivity of both L. albus and L. angustifolius.     

Technological review of the HRP manufacturing process R&D activity

ENEA and Ansaldo Nucleare S.p.A. have been deeply involved in the European International Thermonuclear Experimental Reactor (ITER) R&D activities for the manufacturing of high heat flux plasma-facing components (HHFC), and in particular for the inner vertical target (IVT) of the ITER divertor.This component has to be manufactured by using both armour and structural materials whose properties are defined by ITER. Their physical properties prevent the use of standard joining techniques. The reference armour materials are tungsten and carbon/carbon fibre composite (CFC). The cooling pipe is made of copper alloy (CuCrZr-IG).During the last years ENEA and Ansaldo have jointly manufactured several actively cooled monoblock mock-ups and prototypical components of different length, geometry and materials, by using innovative processes: HRP (hot radial pressing) and PBC (pre-brazed casting).The history of the technical issues solved during the R&D phase and the improvements implemented to the assembling tools and equipments are reviewed in the paper together with the testing results.The optimization of the processes started from the successful manufacturing of both W and CFC armoured small scale mockups thermal fatigue tested in the worst ITER operating condition (20 MW/m²) through the achievement of record performances obtained from a monoblock medium scale mockup.On the base of these results ENEA-ANSALDO participated to the European programme for the qualification of the manufacturing technology to be used for the procurement of the ITER divertor IVT, according to the F4E specifications. A divertor inner vertical target prototype (400 mm total length) with three plasma facing component units, was successfully tested at ITER relevant thermal heat fluxes.Now, ANSALDO and ENEA are ready to face the challenge of the ITER inner vertical target production, transferring to an industrial production line the experience gained in the development, optimization and qualification of the PBC and HRP processes.     

Inactivation and reactivation of horseradish peroxidase treated with supercritical carbon dioxide

Effects of supercritical carbon dioxide (SCCO₂) on the activity of horseradish peroxidase (HRP) in pH 5.6 acetate buffer solution were investigated. SCCO₂ treatment could effectively inactivate HRP. Higher pressure, higher temperature, and longer treatment time caused more inactivation. The maximum reduction of HRP activity reached nearly 90% at 30 MPa and 55 °C for 60 min. Analysis of first-order reaction kinetic data (characterized by a rate constant k and by a decimal reduction time D) showed that D value was closely related to the pressure and temperature of SCCO₂ treatment. Higher pressures or higher temperatures resulted in lower D values (higher k), the D value of HRP was minimized to 64.52 min treated by the combination of 30 MPa and 55 °C. The Z _p, representing the range of applied pressure between which the D values change by a factor of 10, was 114.81 MPa. The activity of HRP treated by SCCO₂ was reactivated significantly after initial 7-day storage at 4 °C apart from the samples at 30 MPa for 60 min, indicating the HRP inactivation may be reversible and the reactivation of HRP is dependent on the pressure level and treatment time.     

无金属卟啉置换辣根过氧化物酶辅基研究

不同结构的卟啉插入去辅基辣根过氧化物酶(apo-HRP)所得重构酶具有类过氧化物酶的催化活性.采用紫外分光光度法和荧光光度法对这些重构酶进行研究.结果表明TPyP-apo-HRP比其他卟啉类插入重构蛋白具有的类过氧化物酶催化活性高.     

酶标抗原直接竞争ELISA方法检测呋喃唑酮代谢物残留

采用酶标抗原建立了高效、高灵敏的呋喃唑酮代谢物直接竞争的ELISA检测方法。以呋喃唑酮单克隆抗体包被作为固相抗体,HRP标记的抗原与标准品（或样品）中呋喃唑酮的代谢物衍生物竞争结合抗体,建立了直接竞争酶联免疫检测体系。以3-氨基-2-恶唑烷酮的衍生物（CPAOZ）为标准品建立标准曲线,得到方法的IC50为0.08μg/L,灵敏度为0.015μg/L,线性范围0.025⁰.5μg/L;检测样品的平均回收率为82.0%¹21.0%,与其他结构类似物基本无交叉反应。建立呋喃唑酮酶标抗原直接竞争酶联免疫检测方法,灵敏度高、特异性强、操作简单,可以满足畜禽水产实际样品的检测需要。      

Moiety and linker site heterologies for highly sensitive immunoanalysis of cyprodinil in fermented alcoholic drinks

Cyprodinil is a new-generation anilinopyrimidine fungicide widely used in crop protection and frequently found in fruits. In this study, novel derivatives of cyprodinil with linker site heterologies were synthesized and employed in order to produce antibodies with enhanced affinity. Moreover, moiety-heterologous haptens were designed and prepared for assay sensitivity improvement. Two competitive enzyme-linked immunosorbent assays for the analysis of this active substance were developed using direct and indirect formats, achieving IC₅₀ values around 0.15 μg/L. Analytical figures of merit and usability of the optimized assays were evaluated with wine and cider as model food processed matrices. The obtained recoveries were from 90% to 120%, and the limit of quantification was in the 1–5 μg/L range. Finally, a monitoring study (n = 150) was performed to estimate the occurrence and the concentration of cyprodinil in commercial wine and cider products from different origins. We found that 28% of the analysed wine samples contained cyprodinil residues at levels higher than 5 μg/L.     

Size matters: Challenges in imprinting macromolecules

A large number of molecularly imprinted polymers (MIPs) have been investigated and reported over the last decade. Various templates have been successfully exploited and used, leading to significant advances in separation, adsorption, catalysis, sensing, and drug delivery. Among all the templates, small molecules have dominated in the synthesis of MIPs. In contrast, progress made in imprinting macromolecules has been slow, mainly due to the challenges presented by the size, structure and conformational fragility of biological macromolecules. In this review, we focus on discussing some key issues involved in the imprinting of macromolecules from recent publications. The similarity and difference between imprinting small molecules and macromolecules are highlighted. Other aspects relating to polymer design and function are also discussed.     

Effect of a hygienized rennet paste and a defined strain starter on proteolysis,texture and sensory properties of semi-hard goat cheese

A hygienized rennet paste (HRP) and a defined strain starter culture, including Lactobacillus casei subsp. casei IFPL as adjunct, were considered for manufacturing Majorero cheese, a Spanish traditional variety made from goat milk. Influence of both factors on physicochemical characteristics, proteolysis, rheological and sensory properties, was evaluated throughout the ripening. Cheeses produced either industrially (IL) or in artisanal manner (AL) were compared with the experimental lot (EL), which included HRP and IFPL starter in its manufacture. Results showed a low level of primary proteolysis, expressed by a low content of non-casein nitrogen (NCN), in experimental cheeses. Despite the slightly poor texture (hard and crumbly) related to the high TS and salt contents, a good general acceptability was attained for EL, with the best scores for aroma and flavour intensities achieved at 30 ripening days. In fact, the sensory panel detected the “piquant” flavour (typical of the artisanal cheese variety) in EL after 15 days of ripening.     

Humanoid robotics platforms developed in HRP

This paper presents humanoid robotics platform that consists of a humanoid robot and an open architecture software platform developed in METI’s Humanoid Robotics Project (HRP). The final version of the robot, called HRP-2, has 1540 mm height, 58 kg weight and 30 degrees of the freedom. The software platform includes a dynamics simulator and motion controllers of the robot for biped locomotion, falling and getting up motions. The platform has been used to develop various applications and is expected to initiate more humanoid robotics research.     

Biocompatible hybrid film of β-cyclodextrin and ionic liquids: A novel platform for electrochemical biosensing

A novel electrochemical biosensing platform was developed based on the modification of biocompatible hybrid film of β-cyclodextrin (β-CD) and ionic liquids (ILs), 1-butyl-3-methylimidazolium tetrafluoroborate (BMIMBF₄) onto glass carbon electrode (GCE). Then an unmediated biosensor was successfully prepared by immobilizing the horseradish peroxidase (HRP) into the β-CD/ILs composite film. β-CD could provide a biocompatible microenvironment for HRP, and ILs could accelerate the electron transfer between HRP and the electrode. Results showed that the HRP entrapped in the β-CD/ILs film could maintain its native structure and the direct electrochemistry of HRP were facilely achieved. A couple of well-defined redox peaks of HRP were observed at about −0.32 V (vs. SCE), corresponding to the protein heme Fe(III)/Fe(II) redox couples. The electrocatalysis of this biosensor to both quercetin and hydrogen peroxide was characterized. The biosensor exhibited a low operating potential (−0.05 V vs. SCE), fast amperometric response, high sensitivity, good selectivity and sub-micromolar limit of detection.