An investigation of heparinase immobilization

A systematic investigation of the parameters that affect the efficiency of immobilizing heparinase onto cyanogen bromide activated crosslinked 8% agarose beads was conducted. Two experimental measures, the “fraction bound” and the “fraction retained,” were used to monitor the coupling efficiency. The fraction bound is the portion of the total initial enzyme that is bound to the agarose gel. The fraction retained is the fraction of bound enzyme that is active. The product of the two measures indicates the coupling efficiency. The activity of the immobilized heparinase was measured under conditions free of both internal and external mass transfer limitations, and thus, the fraction retained represents the true immobilized enzyme activity. Increasing the degree of activation of the beads results in an increase in the fraction bound, the fraction retained, and consequently, the coupling efficiency. As the ratio of enzyme solution to gel volume increases from 1.5 to 2.2, the fraction bound remains constant but the fraction retained decreases (heparinase concentration; 0.15 mg/mL and degree of activation; 9.5 μmol of cyanate esters/g of gel). At volume ratios greater than 2.2, both the fraction bound and the fraction retained decline continuously. Changing the heparinase concentration in the coupling solution changes the coupling efficiency in a manner similar to that of the volume ratio change. When heparin is added during the coupling process, the fraction bound declines as the heparin concentration increases, whereas the fraction retained increases up to a heparin concentration of 12 mg/mL and decreases thereafter. When arginine, lysine, and glycine are used to block the unreacted cyanate ester groups after the coupling process, the immobilized heparinase shows different pH optima of 6.5, 6.9, and 7.2, respectively. Based upon these findings, a protocol to optimize heparinase immobilization is developed.     

Development of nitride-layer of AISI 304 austenitic stainless steel during high-temperature ammonia gas-nitriding

Ammonia-gas nitriding of AISI 304 austenitic stainless steel was studied at temperatures higher than 800 °C using SEM and X-ray diffraction. The result showed that S-phase, an expanded austenite, was formed even at such high temperatures due to a high nitriding potential of ammonia gas. The equilibrium phase, CrN was formed through a decomposition of S-layer in two different modes; the one was through continuous precipitation of particles at the surface-side of S-layer due to a higher nitriding potential; the other through a discontinuous(-like) precipitation at the austenite interface-side, producing a fine lamellar structure of austenite and CrN. The γ-phase in the surface-side resulting from the precipitation of CrN particles subsequently transformed into Fe₄N because of a fast enrichment of N atoms and a limited mobility of Cr atoms at the surface-side. A coarse lamellar structure made of austenite and Cr₂N was developed in front of fine lamellae composed of austenite and CrN by the decomposition of supersaturated austenite through a discontinuous precipitation via grain boundary movement.     

Application of thiophilic chromatography to deplete serum immunoglobulins in sample preparation for bidimensional electrophoresis

Serum is a typical sample for non-invasive studies in clinical research. Its proteome characterization is challenging, since requires extensive protein depletion. Methods used nowadays for removal of high-abundance proteins are expensive or show quite often a low loading capacity, which has strong repercussions on the number of samples and replicates per analysis.In order to deplete immunoglobulins (Igs) and albumin (HSA) from 1 mL serum samples, we have developed a protocol based on a combination of thiophilic chromatography, not previously used in clinical proteomics, and a HSA-specific resin. Ig/HSA-depleted samples, immunoglobulinome and albuminone were analyzed by 2-DE. Thiophilic chromatography, coupled with HSA-depletion, allows a good 2-DE resolution as well as the visualization of new spots. Moreover, it yields enough protein to evaluate technical variability and facilitate subsequent protein identification. To validate the protocol, we carried out a preliminary comparative study between triplicate Igs/HSA-depleted serum samples from healthy control individuals and recently diagnosed/untreated rheumatoid arthritis (RA) patients. RA patients showed several acute phase proteins, as well as additional serum proteins, differentially and significantly regulated.Therefore, thiophilic chromatography can be used as an efficient and economical method in 2-DE to deplete immunoglobulins from large human serum samples before a more extensive fractioning.     

收入管理的MMV动态拍卖机制研究

随着互联网上交易的增多,多单位同质产品销售的拍卖机制成为了一个新的研究方向.针对易逝性服务产品的收入管理问题,提出将产品销售分为多个拍卖期的MMV(Multi-period and Multi-unit Vickrey(Auction with Reserve Pricing)动态拍卖机制,给出每期最优拍卖单位数和保留价格的确定方法,证明了最优拍卖单位数分别关于剩余存量和销售时间单增的性质,以及保留价格关于剩余存量的单降性.最后证明MMV机制的每期实际成交价格将高于Vulcano(2002)提出的MSP(Modified Second-Price)机制,根据易逝性服务产品的需求特点得到MMV机制优于MSP机制的结论.     

Effect of shot peening on the microstructure of laser hardened 17-4PH

In order to investigate the influence of shot peening on microstructure of laser hardened steel and clarify how much influence of initial microstructure induced by laser hardening treatment on final microstructure of laser hardened steel after shot peening treatment, measurements of retained austenite, measurements of microhardness and microstructural analysis were carried out on three typical areas including laser hardened area, transitional area and matrix area of laser hardened 17-4PH steel. The results showed that shot peening was an efficient cold working method to eliminate the retained austenite on the surface of laser hardened samples. The surface hardness increased dramatically when shot peening treatments were carried out. The analyses of microstructure of laser hardened 17-4PH after shot peening treatment were carried out in matrix area and laser hardened area via Voigt method. With the increasing peening intensity, the influence depth of shot peening on hardness and microstructure increased but the surface hardness and microstructure did not change when certain peening intensity was reached. Influence depth of shot peening on hardness was larger than influence depth of shot peening on microstructure due to the kinetic energy loss along the depth during shot peening treatment. From the microstructural result, it can be shown that the shot peening treatment can influence the domain size and microstrain of treated samples but laser hardening treatment can only influence the microstrain of treated samples.     

The austenite microstructure evolution in a duplex stainless steel subjected to hot deformation

Abstract

The austenite microstructure evolution and softening processes have been studied in a 23Cr–6Ni–3Mo duplex stainless steel, comprising equal fractions of austenite and ferrite, deformed in uniaxial compression at 1000 °C using strain rates of 0.1 and 10 s^?1. The texture and microstructure evolution within austenite was similar in character for both the strain rate used. The observed large-scale subdivision of austenite grains/islands into complex-shaped deformation bands, typically separated by relatively wide transition regions, has been attributed to the complex strain fields within this phase. Organised, self-screening microband arrays were locally present within austenite and displayed a crystallographic character for a wide range of austenite orientations. The microband boundaries were aligned with the traces of {1?1?1} slip planes containing slip systems having high, although not necessarily the highest possible, Schmid factors. The slightly lower mean intercept length and higher mean misorientation obtained for the sub-boundaries at the higher strain rate can be ascribed to the expected more restricted dynamic recovery processes compared to the low strain rate case. Dynamic recrystallisation within austenite was extremely limited and mainly occurred via the strain-induced migration of the distorted original twin boundaries, followed by the formation of multiple twinning chains.     

Fe—C—Cr—Mn亚稳奥氏体铸铁磨损表层的TEM观察

用透射电镜观察了 Fe- C- Cr- Mn亚稳奥氏体基铸铁在 80 0 #Si C砂纸上经往复摩擦磨损试验后磨损表层的微观结构 .结果表明 :摩擦表层奥氏体基体的微观组织结构不均匀 ,有位错区、层错区、滑移区和ε-马氏体区 ;此外 ,还观察到了大量的奥氏体变体 ,沿其 [1 31 ]晶带轴旋转 2 5.35°可与基体 [1 1 2 ]晶带轴重合 ;在碳化物颗粒附近只观察到奥氏体变体和层错 .     

稀土奥贝蠕铁制取工艺及其性能的研究

稀土奥贝蠕铁可用含稀土的蠕化剂及等温淬火工艺稳定地制取,藉助高温扫描电子显微镜,观察并研究了稀土蠕铁奥氏体化过程及稀土奥贝蠕铁随温度升高时组织的变化。用带有加热装置的万能材料试验等土奥贝蠕铁的各种性能进行了系统测定了如抗拉强度、延伸率、冲吉韧性、热膨胀系数及伸长百分率,结果表明,稀土奥贝蠕铁在室温和高温下均有良好的性能。     

残余奥氏体含量对马氏体高强钢滑动磨损性能的影响

对3种不同残奥(RA)含量的马氏体高强钢进行干滑动摩擦磨损试验, 研究RA含量对其磨损性能的影响. 利用扫描电镜、透射电镜、X射线衍射仪等对试验后的磨损表面及横截面显微组织进行表征. 结果表明, RA含量越高, 磨损表面越光滑, 摩擦系数和磨损率越小, 也即马氏体高强钢的耐磨性越好. 磨损引起的大应变使RA发生应变诱导马氏体相变, 导致硬度和硬化层厚度显著增大. RA含量最高的HT3试样的硬度提高了18.3%, 硬化层厚度达70μm. 相比RA含量低的试样, HT3试样表现出很好的耐磨性. 这是因为马氏体相变使硬度逐步增加, 抗裂纹萌生能力提高; 同时由于亚表面良好的韧性, 可延缓和阻止裂纹扩展, 使得点蚀和剥落不易形成. 因此, 要提高马氏体高强钢的耐磨性, 除了硬度要求外, 还需要考虑其亚表面韧性.