Proteinase pattern of honeybee prepupae from healthy and American Foulbrood infected bees investigated by zymography

American foulbrood disease (AFB) is the main devastating disease that affects honeybees’ brood, caused by Paenibacillus larvae. The trend of the research on AFB has addressed the mechanisms by which P. larvae bacteria kill honeybee larvae. Since prepupae could react to the infection of AFB by increasing protease synthesis, the aim of this work was to compare protease activity in worker prepupae belonging to healthy colonies and to colonies affected by AFB. This investigation was performed by zymography. In gel, proteolytic activity was observed in prepupae extracts belonging only to the healthy colonies. In the prepupae extracts, 2D zimography followed by protein identification by MS allowed to detect Trypsin‐1 and Chymotrypsin‐1, which were not observed in diseased specimens. Further investigations are needed to clarify the involvement of these proteinases in the immune response of honeybee larvae and the mechanisms by which P. larvae inhibits protease production in its host.     

竹红菌素的结构修饰

竹红菌甲素(简称甲素,HA)是一种新发现的苝醌衍生物,可作光疗药物和光敏化剂。为了筛选出更好的光疗药物,我们逐步修饰了竹红菌素的结构。痂囊腔菌素1的2位和11位甲氧基可被三氯化铝选择性地脱去甲基,而6位和7位的甲氧基不变化,但反应后产物难于分离提     

Effects of diffusion of a naturally-derived plasticizer from soft PVC

Soft PVC was obtained by using a new plasticizer, based on cardanol, a renewable resource characterized by chemical and physical properties very close to those of diethylhexyl phthalate (DEHP). Cardanol acetate (CA) was obtained by solvent free esterification of cardanol, and used as secondary plasticizer, by partial substitution of DEHP in soft PVC formulations. Ageing tests were performed in order to study the stability of properties of the soft PVC formulations related to plasticizer diffusion. Tensile properties and hardness changes were used to monitor the macroscopic effects of plasticizer diffusion. Soft PVC obtained by partial substitution of DEHP by CA showed a significant modification of mechanical properties related to a higher plasticizer evaporation during ageing tests. Migration tests confirmed that CA is characterized by a higher diffusivity in soft PVC compared to DEHP.     

Preparation of Hybrid Nanocapsules

Hybrid nanoparticles with a polystyrene core and a hybrid copolymer shell were used to produce hybrid nanocapsules by dissolving the polystyrene core from the previously elaborated core-shell particles. Following previous works, the core-shell particles were prepared by emulsion polymerization of styrene and subsequent addition of γ-methacryloxy propyl trimethoxy silane (MPS) to produce the shell by copolymerization reaction of MPS with the residual styrene. Core extraction was performed by diluting the core-shell particles in an excess of tetrahydrofuran (THF). Two procedures were investigated to separate the dissolved polymer chains from the nanocapsules. In the first procedure, the polymer was isolated by successive centrifugation and redispersion in THF, whereas in the second procedure, the free polymer chains were removed by dialysis. The polymer molecular weight was optimized in order to promote dissolution of the polymer chains and allow them to diffuse through the shell.     

Birefringence in polycarbonate: Molecular orientation induced by cooling stresses. II. Effects of sectioning

Birefringence distributions in cross-sections of quenched polycarbonate disks are investigated. The effects of the introduction of new surfaces by sectioning are studied by using different cross-section widths, by quenching rod-shaped specimens, and by measuring the birefringence with oblique incident light. The experimental results are confirmed by numerical calculations of stress distributions in rods with varying aspect ratios. Stress release by appropriate sectioning is used to distinguish between stress and orientation contribution to birefringence. It is found that cutting cross-sections leads to a major stress redistribution in the width direction. This does not affect the birefringence measurement, but the decrease of the stress level in the long direction needs to be taken into account. The molecular orientation in the quenched samples cannot be explained only by the effect of transient thermal stresses above the glass transition. There is another orientation contribution, most likely related to stress relaxation below T_g. © 1993 John Wiley & Sons, Inc.     

Uptake and speciation of selenium in garlic cultivated in soil amended with symbiotic fungi (mycorrhiza) and selenate

The scope of the work was to investigate the influence of selenate fertilisation and the addition of symbiotic fungi (mycorrhiza) to soil on selenium and selenium species concentrations in garlic. The selenium species were extracted from garlic cultivated in experimental plots by proteolytic enzymes, which ensured liberation of selenium species contained in peptides or proteins. Separate extractions using an aqueous solution of enzyme-deactivating hydroxylamine hydrochloride counteracted the possible degradation of labile selenium species by enzymes (such as alliinase) that occur naturally in garlic. The selenium content in garlic, which was analysed by ICP–MS, showed that addition of mycorrhiza to the natural soil increased the selenium uptake by garlic tenfold to 15 μg g⁻¹ (dry mass). Fertilisation with selenate and addition of mycorrhiza strongly increased the selenium content in garlic to around one part per thousand. The parallel analysis of the sample extracts by cation exchange and reversed-phase HPLC with ICP–MS detection showed that γ-glutamyl-Se-methyl-selenocysteine amounted to 2/3, whereas methylselenocysteine, selenomethionine and selenate each amounted to a few percent of the total chromatographed selenium in all garlic samples. Se-allyl-selenocysteine and Se-propyl-selenocysteine, which are selenium analogues of biologically active sulfur-containing amino acids known to occur in garlic, were searched for but not detected in any of the extracts. The amendment of soil by mycorrhiza and/or by selenate increased the content of selenium but not the distribution of detected selenium species in garlic. Finally, the use of two-dimensional HPLC (size exclusion followed by reversed-phase) allowed the structural characterisation of γ-glutamyl-Se-methyl-selenocysteine and γ-glutamyl-Se-methyl-selenomethionine in isolated chromatographic fractions by quadrupole time-of-flight mass spectrometry.     

Comparison of solution-blending and melt-intercalation for the preparation of poly(ethylene-co-acrylic acid)/organoclay nanocomposites

Ethylene-acrylic acid copolymers (EAAs) and commercial montmorillonite clays organically modified with dimethyldihydrogenatedtallowammonium ions (Cloisite^® 15A and 20A) were used for the synthesis of nanocomposites by melt-compounding, static melting of polymer/clay mixtures and solution-intercalation in order to compare the effectiveness of these procedures and to shed light on the thermodynamics and the kinetics of the intercalation process. The preparation from solution was made by the use of several solvents, such as toluene, xylene, chloroform, etc., which were then removed from the hybrids by precipitation in different non-solvents or by evaporation. Particular attention was paid to the effect of the thermal treatments which are often used when processing the composites prepared from solution. X-ray diffraction (XRD) of the solution-blended composites showed that no intercalation of the EAAs inside the clay galleries can be achieved if solvent removal is made by precipitation in non-solvents or by room-temperature evaporation. On the contrary, intercalation was found to occur very rapidly (in less than 1 min) when both the hybrids prepared from solution and the mechanical blends of powdered components were melted in the absence of shear. Polymer intercalation was also found to occur, though with a lower rate, upon annealing the powder mixtures at temperatures lower than the EAA melting point. Microscopic observations made by polarized optical microscopy, scanning electron microscopy and transmission electron microscopy showed that the clay particles dispersion is appreciably lower for the composites prepared from solution, compared to those produced in the melt under shear flow conditions. The hybrids obtained by static melting of powder mixtures, on the other side, were expectedly found to comprise micron sized clay agglomerates, although intercalation was demonstrated also for these materials by XRD. The structure of the intercalated silicate layers stacks, characterized by an interlayer spacing of 4.0 nm, was shown to be independent of the preparation procedure and to correspond to thermodynamic equilibrium.     

纳米羟基磷灰石/胶原复合材料制备方法比较研究

低温下,通过将水热合成的纳米羟基磷灰石浆料与中性胶原溶胶共混和在中性胶原中原位形成羟基磷灰石两种方法制备羟基磷灰石／胶原复合材料,采用XRD、FTIR、扫描电镜、透射电镜和力学性能测试等方法对两种复合材料的特性进行了表征。通过对两种方法制备的复合材料的特性进行比较,发现两种方法均制备得到了纳米羟基磷灰石／胶原复合材料,复合材料在晶相组成、化学组成、纳米羟基磷灰石晶体尺寸、胶原纤维的结构等方面都与天然骨相似。但原位合成纳米羟基磷灰石晶体的结晶度比水热合成的纳米羟基磷灰石更接近于自然骨,原位合成的羟基磷灰石／胶原复合材料的均匀性、界面结合紧密度、力学性能等方面均优于共混法。原位合成法是改善纳米羟基磷灰石／胶原复合材料均匀性和力学性能的有效方法。     

A mini-review and perspective on ferroptosis-inducing strategies in cancer therapy

This review summarized recent advances and challenges in ferroptosis-based anticancer strategies through Fenton reaction or GPX4 inactivation, with highlights on nanomaterials and perspectives on future development of next-generation ferroptosis-inducing agents based on diverse polyvalent metals.     

Improving the knowledge and design of end groups in polymers produced by free radical polymerization

Several techniques have been used to probe polymer end groups. The nitroxide radical trapping technique has been used (i) to show that initiator-derived unsaturated end groups in polymethyl methacrylate can be minimized by using t-hexyl peroxypivalate as the initiator (ii) to predict the end and penultimate groups in acrylonitrile/ethyl vinyl ether copolymer produced by t-butoxyl initiation by analogy with the initiation mechanism (iii) to predict probable end groups in polyacrylonitrile and polystyrene produced by cyanoisopropyl initiation in the presence of adventitious oxygen. NMR techniques have been used to show that the end groups of functionalized oligomers, made from styrene and methacrylonitrile by the addition-fragmentation chain transfer technique with allylic sulphides, conform to the expected structures. © 1998 John Wiley & Sons, Ltd.     

乙烷在金属铁表面还原NO的实验研究

温度300~1 100 ℃时,由程序控温电加热水平陶瓷管反应器在N₂气氛和模拟气氛下,对乙烷在金属铁表面还原NO的特性进行了实验研究。结果表明,乙烷在金属铁表面能够高效地还原NO。在N₂气氛中,温度高于900 ℃时,乙烷在金属铁表面的脱硝效率超过95%。在模拟烟气条件下,当温度超过900 ℃,且过量空气系数小于1.0时,乙烷在金属铁表面还原NO的效率能够达到90%以上。相同条件下,乙烷在金属铁表面脱硝效率高于甲烷的脱硝效率。SO₂对乙烷在金属铁表面还原NO的效率影响可以忽略。对反应后的铁样品的组分进行了XRD表征,在此基础上对反应机理进行了分析。结果表明,在模拟烟气条件下NO的还原通过乙烷的再燃脱硝和金属铁直接还原两个机理完成。金属铁直接还原NO时生成的氧化铁则被乙烷还原为金属铁,从而使得金属铁能够持续对NO进行直接还原。乙烷再燃还原NO的中间产物HCN被氧化铁氧化为N₂,同时氧化铁也被HCN还原为金属铁。这一过程增强了NO的持续还原反应,同时避免了在燃尽时HCN二次氧化重新生成NO,从而保证了较高的NO还原效率。     

Exfoliation of layered double hydroxides by use of zwitterionic surfactants in aqueous solution

A new strategy to exfoliate inorganic layered double hydroxide (LDH) solids to micrometer-sized and positively-charged nanosheets in aqueous solution is presented. The procedure involves intercalation of zwitterionic surfactant into the decarbonated LDH by ion exchange, followed by simply stirring the surfactant-intercalated LDH in an aqueous solution of pH?=?2. Since the charge of zwitterionic surfactant can be varied from anionic to cationic by adjusting the pH, the zwitterionic ions present in the interlayer were converted to cationic at pH?=?2, resulting in the exfoliation of LDH by electrostatic repulsion. The delaminated LDHs nanosheets were confirmed by XRD, TEM, and AFM analyses. This simple method did not need reflux at high temperature, overcame the drawback of using organic solvents, and even resulted in a stably colloidal dispersion of nanosheets.     

A pyridoxal-based chemosensor for visual detection of copper ion and its application in bioimaging

A pyridoxal-based chemosensor was synthesized by reacting hydrazine hydrate and pyridoxal hydrochloride in ethanol and characterized by NMR and ESI-MS.The optical properties of the compound were investigated in a methanol:HEPES solution.The compound displayed selectivity for Cu2+,as evidenced by a colorless to yellow color change,which was characterized using UV–vis spectroscopy.The fluorescence of the compound can be quenched only by Cu2+,accompanying by a color change from blue to colorless.Furthermore,it can be used in bioimaging.     

Papain Purification Insights: Monitoring by Electrophoretic Approaches and MALDI-TOF Peptide Mass Fingerprint Analyses

Papain was purified from dried Carica papaya latex by fractioned salt precipitation in presence of sodium tetrathionate to preserve enzymatic activity. Purification was followed by different electrophoretic methods. Identification of the purified product was afforded by submitting the peptides obtained by tryptic digestion of papain to matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF/TOF MS) analysis. Comparison of the peptide masses analyzed by peptide mass fingerprinting (PMF) MALDI-TOF and those obtained by theoretical tryptic digestion, revealed the presence of some peptides belonging the other three endopeptidases contained in papaya latex (very similar to papain in molecular weight and pI) in the purified fraction of papain. The PMF by MALDI-TOF could be applied as a method to follow papain purification.     

Plasma brominated polymer particles as grafting substrate for thiol‐terminated telomers

A combined surface activation and “grafting to” strategy was developed to convert divinylbenzene particles into weak cation exchangers suitable for protein separation. The initial activation step was based on plasma modification with bromoform, which rendered the particles amenable to further reaction with nucleophiles by introducing Br to a surface content of 11.2 atom‐%, as determined by X‐ray photoelectron spectroscopy. Grafting of thiol‐terminated glydicyl methacrylate telomers to freshly plasma activated surfaces was accomplished without the use of added initiator, and the grafting was verified both by reduction in bromine content and the appearance of sulfur‐carbon linkages, showing that the surface grafts were covalently bonded. Following grafting the attached glydicyl methacrylate telomer tentacles were further modified by a two‐step procedure involving hydrolysis to 2,3‐hydroxypropyl groups and conversion of hydroxyl groups to carboxylate functionality by succinic anhydride. The final material was capable of baseline separating four model proteins in 3 min by gradient cation exchange chromatography in a fully aqueous eluent.     

Mechanical Deformation Accelerates Protein Ageing

A hallmark of tissue ageing is the irreversible oxidative modification of its proteins. We show that single proteins, kept unfolded and extended by a mechanical force, undergo accelerated ageing in times scales of minutes to days. A protein forced to be continuously unfolded completely loses its ability to contract by folding, becoming a labile polymer. Ageing rates vary among different proteins, but in all cases they lose their mechanical integrity. Random oxidative modification of cryptic side chains exposed by mechanical unfolding can be slowed by the addition of antioxidants such as ascorbic acid, or accelerated by oxidants. By contrast, proteins kept in the folded state and probed over week‐long experiments show greatly reduced rates of ageing. We demonstrate a novel approach whereby protein ageing can be greatly accelerated: the constant unfolding of a protein for hours to days is equivalent to decades of exposure to free radicals under physiological conditions.     

β-环糊精构筑氨基酸氧化酶的研究

报道了用β-环糊精二间羧基苯磺酸酯与三氯化铁形成的配合物的构筑模拟氨基酸氧化酶, 将苯丙氨酸氧化脱氨生成苯丙酮酸, 在过量H2O2的存在下, 苯丙酮酸进一步被氧化脱羧生成苯乙酸, 若此反应体系是在N, N-二甲基甲酰胺(以下缩写DMF)的弱碱性条件下, 反应则是将苯氨酸氧化成酪氨酸。     

Long-term performance of poly(vinyl chloride) cables. Part 1: Mechanical and electrical performances

Cables insulated with plasticized poly(vinyl chloride) were aged in air at temperatures between 80 °C and 180 °C and their conditions were assessed by indenter modulus measurements, tensile testing, infrared (IR) spectroscopy and differential scanning calorimetry (DSC). Electrical testing of oven-aged cable samples was performed in order to relate the electrical functionality during a high-energy line break (HELB) to the mechanical properties and to establish a lifetime criterion. The mechanical data taken at room temperature after ageing could be superimposed with regard to ageing time and temperature. The ageing-temperature shift factor showed an Arrhenius temperature dependence. The jacketing material showed an immediate increase in stiffness (indenter modulus and Young's modulus) and a decrease in the strain at break on ageing; these changes were dominated by loss of plasticizer by migration which was confirmed by IR spectroscopy and DSC. The core insulation showed smaller changes in these mechanical parameters; the loss of plasticizer by migration was greatly retarded by the closed environment, according to data obtained by IR spectroscopy and DSC, and the changes in the mechanical parameters were due to chemical degradation (dehydrochlorination). A comparison of data obtained from this study and data from other studies indicates that extrapolation of data for the jacketing insulation can be performed according to the Arrhenius equation even down to service temperatures (20-50 °C). The low-temperature deterioration of the jacketing is, according to this scheme, dominated by loss of plasticizer by migration.     

Highly efficient proteome analysis with combination of protein pre-fractionation by preparative microscale solution isoelectric focusing and identification by μRPLC-MS/MS with serially coupled long microcolumn

To improve the efficiency of proteome analysis, a strategy with the combination of protein pre-fractionation by preparative microscale solution isoelectric focusing, peptide separation by μRPLC with serially coupled long microcolumn and protein identification by ESI-MS/MS was proposed. By preparative microscale solution isoelectric focusing technique, proteins extracted from whole cell lysates of Escherichia coli were fractionated into five chambers divided by isoelectric membranes, respectively with pH range from 3.0 to 4.6, 4.6 to 5.4, 5.4 to 6.2, 6.2 to 7.0 and 7.0 to 10.0. Compared to the traditional on-gel IFF, the protein recovery could be obviously improved to over 95%. Subsequently, the enriched and fractionated proteins in each chamber were digested, and further separated by a 30-cm long serially coupled RP microcolumn. Through the detection by ESI-MS/MS, about 200 proteins were identified in each fraction, and in total 835 proteins were identified even with one-dimensional μRPLC-MS/MS system. All these results demonstrate that by such a combination strategy, highly efficient proteome analysis could be achieved, not only due to the in-solution protein enrichment and pre-fractionation with improved protein recovery but also owing to the increased separation capacity of serially coupled long μRPLC columns.     

ICP-MS测量环境样品中铀的非质谱干扰内标校正研究

内标的合理应用直接关系到测量结果的可靠性,通过外标与内标校正相结合的方法,实验研究了Y,Rh,In,Tb,Yb,Tl,Bi,Re作为内标对ICP-MS测量铀的非质谱干扰的校正效果。对于非抑制/增强基体效应,各内标元素校正结果明显优于无内标校正结果,高质量数内标元素Tb,Yb,Re,Tl,Bi可获得较好的校正效果,校正偏差优于5%,其中以Tl为最佳;对于盐度较高的样品溶液,由于基体对待测元素和内标元素产生了明显不同的抑制/增强效应,因此各内标校正结果均不理想,这表明内标对可溶性固体引起的抑制/增强效应的校正能力有限。在铀的测量中,对于含盐度在300～500μg/mL的样品,内标校正造成的偏差可达到5%～10%;对于盐度在500μg/mL以上的样品,内标校正造成的偏差很可能超过10%或者更高。