Myelinosome Organelles in the Retina of R6/1 Huntington Disease (HD) Mice: Ubiquitous Distribution and Possible Role in Disease Spreading

Visual deficit is one of the complications of Huntington disease (HD), a fatal neurological disorder caused by CAG trinucleotide expansions in the Huntingtin gene, leading to the production of mutant Huntingtin (mHTT) protein. Transgenic HD R6/1 mice expressing human HTT exon1 with 115 CAG repeats recapitulate major features of the human pathology and exhibit a degeneration of the retina. Our aim was to gain insight into the ultrastructure of the pathological HD R6/1 retina by electron microscopy (EM). We show that the HD R6/1 retina is enriched with unusual organelles myelinosomes, produced by retinal neurons and glia. Myelinosomes are present in all nuclear and plexiform layers, in the synaptic terminals of photoreceptors, in the processes of retinal neurons and glial cells, and in the subretinal space. In vitro study shows that myelinosomes secreted by human retinal glial Müller MIO-M1 cells transfected with EGFP-mHTT-exon1 carry EGFP-mHTT-exon1 protein, as revealed by immuno-EM and Western-blotting. Myelinosomes loaded with mHTT-exon1 are incorporated by naive neuronal/neuroblastoma SH-SY5Y cells. This results in the emergence of mHTT-exon1 in recipient cells. This process is blocked by membrane fusion inhibitor MDL 28170. Conclusion: Incorporation of myelinosomes carrying mHTT-exon1 in recipient cells may contribute to HD spreading in the retina. Exploring ocular fluids for myelinosome presence could bring an additional biomarker for HD diagnostics.     

重离子束诱变呼吸缺陷型酵母菌株的筛选与快速鉴定

利用单核能为5.19 MeV/u的22Ne5 辐照啤酒酵母菌株,采用TTC筛选培养基筛选出呼吸缺陷型酵母菌株。通过一种新型而简便的限制性酶切分析手段,有效地对所筛选出的8株呼吸缺陷型酵母菌株进行遗传学鉴定。     

红曲发酵液中水溶性红曲黄色素的吸附分离

本文考察不同极性大孔树脂、离子交换树脂、活性炭及硅胶等吸附介质对红曲菌深层发酵液中水溶性红曲黄色素的吸附分离性能。结果显示非极性大孔树脂DA201-C效果最好,该吸附过程符合Freundlich方程,吸附动力学符合准一级动力学模型,表明吸附不受单层吸附的限制;液膜扩散是吸附主要限速步骤,扩散速率常数为0.16 min-1。通过静态和动态吸附洗脱条件优化,运用SPSS对数据进行分析,发现静态吸附、解吸料液比为1:2,吸附、解吸时间15 min,解吸液100%乙醇,酸性条件解吸效果较好;动态吸附解吸流速为0.25 BV/min条件下,可得到高回收率（96.99%）、脱盐率（99.44%）和脱糖率（92.52%）的水溶性红曲黄色素,经初步扩大实验显示吸附解吸性能稳定。大孔树脂DA201-C分离纯化发酵液中水溶性红曲黄色素操作简单、快速、放大性能好,具有很好的工业化生产应用价值。     

红光对紫色红曲霉生长及色素和桔霉素产量的影响

本文研究了红光对紫色红曲霉M9生长及色素和桔霉素产量的影响。采用高效液相色谱法对六种红曲色素,包括两种红色素：红斑红曲胺（rubropunctamine,RUM）和红曲玉红胺（monascorubramine,MOM）;两种黄色素：红曲素（monascin,MS）和红曲黄素（ankaflavin,AK）;两种橙色素：红斑红曲素（rubropunctatin,RUN）和红曲玉红素（monascorubrin,MON）以及桔霉素的产量进行测定。结果表明：持续红光照射促进了紫色红曲霉M9菌丝生长、色素合成积累以及孢子形成,尤其对于闭囊壳的产生有更显著促进作用。5种不同的红光照射条件促进了RUM、MOM、MS和AK的产生,抑制了RUN、MON和桔霉素的产生。在光照强度300 lux,光照时间30 min/d,光照节律12 h的最佳光照条件下,RUM、MOM的产量分别提高了53.8%和75.2%;MS和AK的产量分别提高了42.2%和59.4%;RUN和MON的产量分别降低了42.6%和54.5%;桔霉素的产量降低了42.5%。     

红曲霉菌体残渣酶解条件优化研究

为提高红曲霉菌（Monascus purpureus）体残渣的利用率,采用机械破壁和酶解结合方法释放胞内蛋白。细胞破碎液经纤维素酶与复合酶联合处理,通过正交试验确定最佳的水解条件为红曲霉菌体残渣质量浓度60 g/L、酸性纤维素酶用量1.0%、复合酶用量0.5%、水解时间36 h。在此条件下,红曲霉菌体残渣的失重率达到27.86%,总糖含量达到7.38 g/L,小肽含量达到83.95 mg/g,游离氨基酸含量达到22.31 mg/g。     

保加利亚乳杆菌与嗜热链球菌抗噬菌体菌株的原生质体制备及融合

为获得具有抗噬菌体功能且发酵性能优良的乳酸菌融合子,采用单亲灭活及正交分析方法,研究了保加利亚乳杆菌与嗜热链球菌抗噬菌体菌株的原生质体制备及融合条件。结果表明：保加利亚乳杆菌原生质体制备的最适条件是以磷酸盐缓冲液和甘露醇制作的高渗溶液为原生质体稳定剂,1.0 mg/mL的溶菌酶36 ℃处理30 min,原生质体的形成率为（89.02±2.31）%,再生率为（4.62±0.22）%。嗜热链球菌抗噬菌体菌株原生质体制备的最适条件是以Tris-HCl和蔗糖制作的高渗溶液为原生质体稳定剂,0.1 mg/mL的溶菌酶42 ℃处理30 min,原生质体的形成率为（99.15±0.23）%,再生率为（5.79±0.17）%。单亲灭活保加利亚乳杆菌与嗜热链球菌抗噬菌体菌株原生质体融合的最适条件为聚乙二醇6000（质量浓度为400 g/L,添加0.01 mol/L CaCl2、0.02 mol/L MgCl2）40 ℃促融2 min,融合率可达（1.85±0.12）×10－6。所得融合子各项性能优良,适合于酸奶生产。     

MsiR高可溶性蛋白突变株的筛选

外源蛋白在表达宿主中的可溶性是影响其应用的关键因素。采用mCherry 报告基因,构建MsiR-mCherry融合蛋白,通过测定荧光值定性定量的测定MsiR蛋白在大肠杆菌中的可溶性表达量。随后,利用易错聚合酶链反应（Errorprone-PCR）和致突变菌株（XL1-Red）两种方式构建了msiR-mCherry 基因突变文库,经筛选得到了4株在大肠杆菌中以37 ℃作为诱导温度下可溶性明显提高的蛋白突变体。     

Molecular Dynamics Studies on the Structural Characteristics for the Stability Prediction of SARS-CoV-2

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) affects the COVID-19 pandemic in the world. The spike protein of the various proteins encoded in SARS-CoV-2 binds to human ACE2, fuses, and enters human cells in the respiratory system. Spike protein, however, is highly variable, and many variants were identified continuously. In this study, Korean mutants for spike protein (D614G and D614A-C terminal domain, L455F and F456L-RBD, and Q787H-S2 domain) were investigated in patients. Because RBD in spike protein is related to direct interaction with ACE2, almost all researches were focused on the RBD region or ACE2-free whole domain region. The 3D structure for spike protein complexed with ACE2 was recently released. The stability analysis through RBD distance among each spike protein chain and the binding free energy calculation between spike protein and ACE2 were performed using MD simulation depending on mutant types in 1-, 2-, and 3-open-complex forms. D614G mutant of CT2 domain, showing to be the most prevalent in the global pandemic, showed higher stability in all open-complex forms than the wild type and other mutants. We hope this study will provide an insight into the importance of conformational fluctuation in the whole domain, although RBD is involved in the direct interaction with ACE2.     

Decreasing the cost of mutation testing with second‐order mutants

Although powerful, mutation is a computationally very expensive testing technique. In fact, its three main stages (mutant generation, mutant execution and result analysis) require many resources to be successfully accomplished. Thus, researchers have made important efforts to reduce its costs. This paper represents an additional effort in this sense. It describes the results of two experiments in which, by means of combining the original set of mutants and therefore obtaining a new set of mutants—each one with two faults—the number of mutants used is reduced to half. Results lead to believe that mutant combination does not decrease the quality of the test suite, whereas it supposes important savings in mutant execution and result analysis. Copyright © 2008 John Wiley & Sons, Ltd.     

99.7 Per Cent Pure

The HEPA air filter is only capable of processing 99.7 per cent of airborne particles, leaving 0.3 per cent unaccounted for. Mason White describes the work of Canadian installation artist An Te Liu who made air-filtration appliances his main subject. Copyright © 2009 John Wiley & Sons, Ltd.