Cell-Free Synthesis of Selenoproteins in High Yield and Purity for Selective Protein Tagging

The selenol group of selenocysteine is much more nucleophilic than the thiol group of cysteine. Selenocysteine residues in proteins thus offer reactive points for rapid post-translational modification. Herein, we show that selenoproteins can be expressed in high yield and purity by cell-free protein synthesis by global substitution of cysteine by selenocysteine. Complete alkylation of solvent-exposed selenocysteine residues was achieved in 10 minutes with 4-chloromethylene dipicolinic acid (4Cl-MDPA) under conditions that left cysteine residues unchanged even after overnight incubation. Gd^III−Gd^III distances measured by double electron–electron resonance (DEER) experiments of maltose binding protein (MBP) containing two selenocysteine residues tagged with 4Cl-MDPA-Gd^III were indistinguishable from Gd^III−Gd^III distances measured of MBP containing cysteine reacted with 4Br-MDPA tags.     

Defining Dry Eye from a Clinical Perspective

Over the past decades, the number of patients with dry eye disease (DED) has increased dramatically. The incidence of DED is higher in Asia than in Europe and North America, suggesting the involvement of cultural or racial factors in DED etiology. Although many definitions of DED have been used, discrepancies exist between the various definitions of dry eye disease (DED) used across the globe. This article presents a clinical consensus on the definition of DED, as formulated in four meetings with global DED experts. The proposed new definition is as follows: “Dry eye is a multifactorial disease characterized by a persistently unstable and/or deficient tear film (TF) causing discomfort and/or visual impairment, accompanied by variable degrees of ocular surface epitheliopathy, inflammation and neurosensory abnormalities.” The key criteria for the diagnosis of DED are unstable TF, inflammation, ocular discomfort and visual impairment. This definition also recommends the assessment of ocular surface epitheliopathy and neurosensory abnormalities in each patient with suspected DED. It is easily applicable in clinical practice and should help practitioners diagnose DED consistently. This consensus definition of DED should also help to guide research and clinical trials that, to date, have been hampered by the lack of an established surrogate endpoint.     

NH_3-OH体系中共沉淀法同时回收钕铁硼废料中Nd-Pr-Co-Fe

为了避免回收单一钕铁硼废料中有价元素带来的操作复杂和资源浪费等问题,本研究采用共沉淀法共沉淀出钕铁硼废料中的有价元素Me(Nd,Pr,Co,Fe),制备可用于生产再生钕铁硼的原料;根据质量守恒和同时平衡原理,采用MATLAB软件建立Me(Nd,Pr,Co,Fe)-OH--NH3热力学模型,绘制lg[Me]-p H曲线模拟共沉淀工艺,并根据模拟结果确立了共沉淀工艺;模拟和实验的结果表明:根据lg[Me]-p H模拟结果可以确立一步共沉淀法的p H:6~10,Fe3+比Fe2+更易于沉淀完全;在上述条件下获得的共沉淀粉末主相均为Nd,Pr,Co,Fe的化合物,且有价元素的百分比含量均大于99.4%;其中,当p H值在8左右时回收率最高,在该条件下金属元素Me(Nd,Pr,Co,Fe)的沉淀效率分别为:98.7%,99.9%,93.6%,99.9%。该结果也表明共沉淀法工艺不仅高效,而且所制备的共沉淀粉末可以满足制备二次钕铁硼的需要。