Identification of SYS1 as a Host Factor Required for Shiga Toxin-Mediated Cytotoxicity in Vero Cells

Shiga toxin (STx) or Vero toxin is a virulence factor produced by enterohemorrhagic Escherichia coli. The toxin binds to the glycosphingolipid globotriaosylceramide (Gb3) for its entry, and causes cell death by inhibiting ribosome function. Previously, we performed a loss-of-function screen in HeLa cells using a human CRISPR knockout (KO) library and identified various host genes required for STx-induced cell death. To determine whether this library targeted to the human genome is applicable to non-human primate cells and to identify previously unrecognized factors crucial for STx-induced cell death, we herein performed a similar screen in the African green monkey kidney-derived Vero C1008 subline. Many genes relevant to metabolic enzymes and membrane trafficking were enriched, although the number of enriched genes was less than that obtained in the screening for HeLa cells. Of note, several genes that had not been enriched in the previous screening were enriched: one of these genes was SYS1, which encodes a multi-spanning membrane protein in the Golgi apparatus. In SYS1 KO Vero cells, expression of Gb3 and sphingomyelin was decreased, while that of glucosylceramide and lactosylceramide was increased. In addition, loss of SYS1 inhibited the biosynthesis of protein glycans, deformed the Golgi apparatus, and perturbed the localization of trans-Golgi network protein (TGN) 46. These results indicate that the human CRISPR KO library is applicable to Vero cell lines, and SYS1 has a widespread effect on glycan biosynthesis via regulation of intra-Golgi and endosome–TGN retrograde transports.     

In Vitro Characterization of Human CD24hiCD38hi Regulatory B Cells Shows CD9 Is Not a Stable Breg Cell Marker

Regulatory B (Breg) cells are endowed with immune suppressive functions. Various human and murine Breg subtypes have been reported. While interleukin (IL)-10 intracellular staining remains the most reliable way to identify Breg cells, this technique hinders further essential functional studies. Recent findings suggest that CD9 is an effective surface marker of murine IL-10 competent Breg cells. However, the stability of CD9 and its relevance as a unique marker for human Breg cells, which have been widely characterized as CD24^hiCD38^hi, have not been investigated. Here, we demonstrate that CD9 expression is sensitive to in vitro B cell stimulations. CD9 expression could either be re-expressed or downregulated in purified CD9-negative B cells and CD9-positive B cells, respectively. We found no significant differences in the Breg differentiation capacity of the CD9-negative and CD9-positive B cells. Furthermore, CD9-positive B cells co-express CD40 and CD86, suggesting their nature as B cell activation or co-stimulatory molecules, rather than regulatory ones. Therefore, we report the relatively unstable CD9 as a distinct surface molecule, indicating the need for further research for a more reliable marker to purify human Breg cells.     

1Cr18Ni9Ti不锈钢氮离子注入层超显微硬度的研究

应用超显微硬度计测定１Ｃｒ１８Ｎｉ９Ｔｉ奥氏体不锈钢氮离子注入层的硬度，并用Ａｕｇｅｒ电子能谱仪（ＡＥＳ）及高精度轮廓仪测定氮离子注入层氮浓度分布及深度．结果表明：在９０ｋｅＶ，３ｘ１０~１７Ｎ＋／ｃｍ~２注入时．共注入层深度为０．１８μｍ、用１ｍＮ载荷侧得本征硬度ＨＶ为３．４７ＧＰａ，比非注入态表面硬度提高三倍之多，合理的试验载荷应≤ｌｍＮ（０．１ｇ）．     

复吹转炉“留渣 双渣”脱磷工艺试验

摘要：在国内某转炉钢厂采用“留渣 双渣”工艺技术进行脱磷工艺试验。结果表明：随着转炉前期脱磷率不断升高,终点脱磷率不断提高。铁水硅含量对前期脱磷率的影响最大。根据铁水成分,在冶炼前期适当降低供氧强度、降低气固氧比、加入适量石灰及烧结矿,均有利于前期脱磷率的提高。在一倒时每吨钢液加入4~8kg石灰,不影响出钢温度,可提高一倒-终点阶段脱磷率,同时可提高终点脱磷率。从终点的控制效果可知,终点炉渣碱度应保持不小于3.0,炉渣中FeO质量分数在16%~20%,并适当降低终点出钢温度在1610~1630℃,有利于终点脱磷率的提高。通过加强熔池搅拌,促进钢渣反应趋于平衡,有利于终点磷分配比提高,从而可进一步提高终点脱磷率。     

Ti9Al合金的氢塑性效应

采用真空自耗电弧炉熔炼制备Ti9Al合金，通过渗氢处理显著提高氢含量，在凸轮塑性试验机ЧПИ-2上以10mm／min的速度进行热模拟试验。研究发现：氢可以降低Ti9Al合金的相变温度，扩大α β区的温度范围；氢作为卢稳定剂会妨碍或完全抑制以Ti3Al为基的α2相的析出；钛合金的氢合金化可促使卢相在更低温度下稳定化，明显地提高了Ti9Al合金塑性。     

磷复肥喷浆造粒“三内”技术研究与开发(一)

介绍磷复肥喷浆造粒“三内”技术 ,该技术集喷浆造粒、分级、返料、破碎为一体在转筒内完成全过程 ,实现机头喷浆机尾出产品 ,粉体在筒内闭式循环造粒新技术     

Effects Of Ozone On The Production Of Biodegradable Dissolved Organic Carbon (BDOC) During Water Treatment

This article deals with the oxidation effect of ozone on the increasing fraction of biodegradable organic matter with the “ozotest” method, a laboratory technique which simulates the effect of ozonation and allows a complete oxidation assessment. Ozone treatment was performed on river water samples and sand filter effluent samples. Ozone consumption, reduction of UV absorbance and BDOC formation were monitored with applied ozone doses from 0 to 10 mg/L and with contact times from 0 to 60 min. The BDOC formation was optimum at an applied ozone dose of 0.25-0.5 mg O₃ per mg DOC (contact time = 5 min) corresponding to apparition of traces of residual ozone and maximum UV reduction. Maximum ozone consumption, UV reduction and BDOC formation occurred simultaneously during the first two minutes of treatment. Concerning BDOC formation, applied ozone dose showed a greater effectiveness than contact time. For the same quantity of consumed ozone, a short contact time associated with a high ozone dose was preferable to a long contact time and a low ozone dose.     

Sex pheromone components of the geometrid mothsLobophora nivigerata andEpirrhoe sperryi

3Z,6Z,9Z-Nonadecatriene (3Z,6Z,9Z-19:H; other abbreviations follow the same pattern) has been identified as a female sex pheromone component of the geometrid mothEpirrhoe sperryi (H.). 3Z,6Z,9Z-18:H and 6Z,9Z-19:H were also identified in pheromone gland extracts but had no apparent biological activity. 3Z,6Z,9Z-21:H was tentatively identified as a female sex pheromone component of a second geometrid species,Lobophora nivigerata (Wlk.). Attraction of male moths to this compound was strongly synergized by the addition of small amounts of 6Z,9Z-21:H to lures.Issued as NRC No. 32478.