Selective labeling of proteins by using protein farnesyltransferase

The challenging task of identifying and studying protein function has been greatly aided by labeling proteins with reporter groups. Here, we present a strategy that utilizes an enzyme that labels a four-residue sequence appended onto the C terminus of a protein, with an alkyne-containing substrate. By using a bio-orthogonal cycloaddition reaction, a fluorophore that carried an azide moiety was then covalently coupled to the alkyne appended on the protein. FRET was used to calculate a F?rster (R) distance of 40 A between the eGFP chromophore and the newly appended Texas Red fluorophore. This experimental value is in good agreement with the predicted R value determined by using molecular modeling. The small recognition tag, the high specificity of the enzyme, and the orthogonal nature of the derivatization reaction will make this approach highly useful in protein chemistry.     

Coexpression of platelet-derived growth factor receptor alpha and fetal liver kinase 1 enhances cardiogenic potential in embryonic stem cell differentiation in vitro

Nascent mesodermal cells derived from EB5 embryonic stem (ES) cells were sorted in terms of cardiogenic potential on the basis of their expression levels of platelet-derived growth factor receptor alpha (PDGFRalpha) and fetal liver kinase 1 (Flk-1). The sorted cells were cocultured with OP9 stromal cells to induce terminal differentiation into contractile cardiac colonies. A significant number of cardiac colonies were found in the Flk-1+/PDGFRalpha+ fraction. The enrichment double-positive fraction produced approximately fivefold more cardiac colonies than the Flk-1+/PDGFRalpha- fraction and 10-fold more than the Flk-1-/PDGFRalpha+ fraction. To investigate the involvement of these markers in embryonic cardiogenesis, the cells that disseminated from the E7.5-7.75 embryos were fractionated and seeded on OP9 cells. The cardiogenic potential was markedly enhanced in the Flk-1+/PDGFRalpha+ fraction. These results suggest that some of the precursor cells coexpressing these markers are selectively involved in cardiogenic events, and that the identification of ES-cell-derived precursors with these markers will contribute to the effective production of cardiomyocytes for cell therapies.     

Characterization of neurons differentiated from mouse embryonic stem cells using conditioned medium of dorsal root ganglia

Mouse embryonic stem (ES) cells have the pluripotent ability to differentiate in vitro into various cell lineages, including neurons. Adding chick dorsal root ganglion (DRG) conditioned medium (CM) to the culture medium promotes the differentiation of ES cells into neurons. We determined the types of neurons that differentiate from ES cells. The addition of DRG-CM caused nearly half of all ES cells on the periphery of the colony sphere to differentiate into neurons. Immunofluorescence analysis showed that the neurons that differentiated from ES cells were mainly motor, GABAergic, serotonergic, and cholinergic neurons. Of particular note, flow cytometry showed that approximately 50% of betaIII-tubulin-positive neurons were motor neurons. This indicates that DRG-CM induces ES cells to differentiate into motor neurons as target of DRG neurons (sensory neurons).     

提高选钼技术经济指标的探讨

介绍了德兴铜矿提高铜钼混合精矿用品位的几项科研成果以及在生产中应用情况。采用充氮新工艺、充填式浮选柱新设备的试验结果表明不仅能提高铜钼分选指标而且可显著降低生产成本。     

氰化工艺过程中金属存量计算方法探讨

为搞好金属平衡，减少理论与实际差值，依据某黄金矿山氰化工艺流程特点，对脱药用单层浓密切、漫出槽、三层洗涤浓密机三个作业中的金属存量的计算方法进行了探讨。     

鲍店煤矿供电安全监测及管理系统的开发

兖矿集团鲍店煤矿应用ＡＵＴＯＣＡＤ、网络技术对分散的供电系统进行改造,实现了采区变电所电网状态及绝缘监测。在地面建立机电管理网,将地面３５ｋＶ 微机监测纳入矿ＷＩＮＤＯＷＳＮＴ 网络,实现了全矿井上下变电所的微机监测和管理,并对井下在线监测做了有益的探索。     

压缩雨季剥离 降低生产成本

本文对露天矿调整采剥进度计划,降低生产成本,提高企业效益作了阐述.     

电子轨道衡转向架计量控制程序的应用

本文论述了转向架计量电子轨道的数据处理过程.对车皮过衡速度、车型判断、重量计算、采集数据的存贮给出了实现方法.此程序适用于绝大多数四轴车以转向架方式过衡.     

Effect of ultrafine grain refinement on hydrogen embrittlement of metastable austenitic stainless steel

Micro-tensile tests were performed on high-pressure-torsion-processed specimens of type 304 steel with grain sizes in the range of 0.1–0.5 μm to clarify the effect of ultrafine grain refinement on the hydrogen embrittlement (HE) of metastable austenitic steel. The ultrafine-grained (UFG) specimens with average grain sizes < ～0.4 μm exhibited a limited uniform elongation followed by a steady-stress regime in the stress–strain curves, which was attributed to a martensitic transformation. A high yield stress and a moderate elongation to failure were attained for the UFG specimens with an average grain size of ～0.5 μm in the uncharged state. Hall–Petch relationships well hold between the yield stress and the average grain size for each uncharged and hydrogen-charged specimen. Hydrogen charging increased the friction stress by 40% but did not change the Hall–Petch coefficient. Hydrogen-induced ductility loss was mitigated by ultrafine grain refinement. Ductility loss due to hydrogen charging manifested in the local deformation after a martensitic transformation. This indicates that hydrogen does not significantly affect the martensitic transformation, but shortens the subsequent local deformation process.     

Contents of eight harmful elements in baby toys and their migration tests

Levels of eight harmful elements, i.e., antimony, arsenic, barium, cadmium, chromium, lead, mercury and selenium, were investigated in 45 baby toys and 10 paints, which were mainly made of polyvinyl chloride. All samples contained barium at levels of 0.3-3,700 mg/kg, several samples contained cadmium (0.2-26 mg/kg), chromium (0.5-280 mg/kg) and lead (1.5-1,300 mg/kg), and one sample contained antimony (5.3 mg/kg). They might have been used as colorants of the toy materials and paints. They were then evaluated using the migration test of ISO 8124-3, in which samples were ground up, and then soaked in 0.07 mol/L HCl at 37 degrees C for two hours. Barium, cadmium, chromium and lead migrated from some of the samples, but at levels lower than the migration limits required by ISO 8124-3. Compared with the Japanese official method, the ISO method resulted in higher migration, but there are significant differences in the migration limits, test method, and so on between them. Further investigation is needed in this area.