PrPC regulates epidermal growth factor receptor function and cell shape dynamics in Neuro2a cells

The prion protein (PrP) plays a key role in prion disease pathogenesis. Although the misfolded and pathologic variant of this protein (PrP^SC) has been studied in depth, the physiological role of PrP^C remains elusive and controversial. PrP^C is a cell‐surface glycoprotein involved in multiple cellular functions at the plasma membrane, where it interacts with a myriad of partners and regulates several intracellular signal transduction cascades. However, little is known about the gene expression changes modulated by PrP^C in animals and in cellular models. In this article, we present PrP^C‐dependent gene expression signature in N2a cells and its implication in the most overrepresented functions: cell cycle, cell growth and proliferation, and maintenance of cell shape. PrP^C over‐expression enhances cell proliferation and cell cycle re‐entrance after serum stimulation, while PrP^C silencing slows down cell cycle progression. In addition, MAP kinase and protein kinase B (AKT) pathway activation are under the regulation of PrP^C in asynchronous cells and following mitogenic stimulation. These effects are due in part to the modulation of epidermal growth factor receptor (EGFR) by PrP^C in the plasma membrane, where the two proteins interact in a multimeric complex. We also describe how PrP^C over‐expression modulates filopodia formation by Rho GTPase regulation mainly in an AKT‐Cdc42‐N‐WASP‐dependent pathway.

     

30 nm染色质的体外组装和电镜分析

真核生物的基因组以染色质的形式存在,染色质在真核生物的基因表达调控及胚胎发育过程中起重要作用,为表观遗传提供一个重要的信息整合平台．染色质的高级结构,特别是 30 nm染色质的动态变化在基因转录沉默和激活过程中起着重要的调控功能．但是目前对30 nm 染色质纤维的组装及其精细结构的认识还十分有限．本文通过体外表达系统,表达未经修饰的组蛋白,并利用克隆构建的601DNA均一重复序列,通过逐步降低盐离子浓度并加入组蛋白H1或镁离子的方法,体外重组均一的30 nm染色质纤维．并利用镀金属、负染色制样和冷冻电镜制样等手段通过透射式电子显微镜(TEM)对30 nm纤维结构的形成原因、组蛋白H1的作用和核小体重复单位(nucleosome repeat lengths,NRLs)长度对30 nm染色质纤维的影响进行研究．研究结果显示在组蛋白H1或二价镁离子存在的情况下,均可形成30 nm染色质纤维．其形成的染色质拓扑结构有所不同．统计分析表明,不同长度核小体重复单位(NRLs)形成的染色质纤维直径有所不同(P < 0.05)．同时,我们得到了较为均一的冷冻电镜样品,为进一步研究30 nm染色质纤维的高级结构及理解体内染色质存在的形式及动态过程打下了较好的基础．     

Role of mitochondrial quality control in exercise-induced health adaptation

Long-term endurance training or physical activity has been confirmed not only to improve physical performance, but to bring about an obvious beneficial effect on human health; however, the mechanism of this effect is not clear. The most studied health adaptations in skeletal muscle response to endurance exercise are increased muscle glycogen level and insulin sensitivity, fiber type transformation toward oxi- dative myofibers, and increased mitochondrial content/function. Mitochondria are dynamic organelles in eukaryotic cells critical in physical performance and disease occurrence. The mitochondrial life cycle spans biogenesis, maintenance, and clearance. Exercise training may promote each of these processes and confer positive impacts on skeletal muscle contractile and metabolic functions. This review focused on the regula- tion of these processes by endurance exercise and discussed its potential benefits in health and disease. We presented evidence suggesting that exercise training potentiates not only the biogenesis of mitochondria but also the removal of old and unhealthy mitochondria through mitochondrial quality control.     

Purification and Properties of Maltose Phosphorylase from Lactobacillus brevis

Maltose phosphorylase (EC 2.4.1.8) from Lactobacillus brevis was purified 29-fold over the crude extract. The final preparation was at least 80% pure and had a specific activity of 18 units/mg protein. The molecular weights of the native enzyme and of the component dissociated in sodium dodecyl sulfate were 150,000 and 80,000, respectively. The enzyme does not contain pyridoxal-5′-phosphate as a cofactor. It can not act on maltitol, malto-triitol, sucrose, lactose and trehalose, and essentially not on isomaltose, maltobionic acid, maltotriose and maltotetraose. Inhibitory effect was observed with CuSO₄, HgCl₂ and p-chloromercuribenzoate. Some other properties were also examined. A possibility of using this enzyme for the analysis of maltose was proposed.     

Effect of 1-n-Decylimidazole on Gibberellin Biosynthesis in Tan-ginbozu,a Dwarf Variety of Rice

Previous structural studies of less-polar dimers in autoxidized methyl linoleate (ML) have been extended to polar dimers. After isolation by successive silicic acid and gel permeation chromatography, the dimeric fraction of linoleate was separated into two major fractions, A₁ and A₂, according to their polarities. The polar dimers (A₁) were further fractionated by HPLC either directly or after reduction with triphenyl phosphine on a micro silica column. Isolated subfractions were characterized by UV, IR, GC-MS and FD-MS after suitable derivatizations. FD-MS of all these dimers showed a molecular ion peak which corresponds to 2 × ML + 6 × O and the reduction of each subfraction with stannous chloride gave equimolar amounts of 9 and 13-hydroxy octadecadienoate, and 9, 10, 13 and/or 9, 12, 13-trihydroxy octadecenoate. These results combined with others show that the A₁ dimers are composed of isomeric mixtures containing a peroxide bridge linking a methyl octadecadienoate and a 9, 12 and/or 10, 13-dihydroperoxy octadecenoate across C-9 and/or 13 on each of them.     

Comparison of the Cathepsin D from Mackerel (Scomber australasicus) and Milkfish (Chanos chanos) Muscle

Muscle proteases from mackerel and milkfish were purified to electrophoretical homogeneity by concanavalin A-Sepharose and Sephadex G-100 chromatographies. Both proteases appear to be an aspartic protease, cathepsin D (EC 3.4.23.5). The molecular weights of the purified cathepsin D’s from mackerel and milkfish were 51,000 and 54,000, estimated by Sephadex G-100, and 59,000 and 61,000 by SDS–PAGE, respectively. Both cathepsin D’s were completely inhibited by pepstatin, but not affected by leupeptin, N-ethylmaleimide, dithiothreitol, or glutathione. ß-Mercaptoethanol, iodoacetic acid, p-chloromercuri-benzoate, phenylmethylsulfonyl fluoride, and sodium dodecyl sulfate partially or completely inhibited both cathepsin D’s. Na⁺ and K⁺ partially activated the cathepsin D from milkfish. Both cathepsin D’s were inhibited by Mg²⁺, Sr²⁺, Fe²⁺, and H²⁺, but activated by Ca²⁺, Co²⁺, Ni²⁺, Cu²⁺, Zn²⁺, and Cd²⁺. The pI and optimal temperature of the cathepsin D’s from mackerel and milkfish were 5.04 and 4.91, 45°, and 50°C, respectively. The temperatures for inactivating 50% activity of the cathepsin D’s from mackerel and milkfish during 20 min of incubation were 53° and 48°C, respectively. Both cathepsin D’s had similar optimal pHs near 3. The activity of that from milkfish markedly decreased when the pH was higher than 4, and was almost completely lost at pH above 6, while that from mackerel still had at least 40% activity at pH 6.     

Very high density of CHO cells in perfusion by ATF or TFF in WAVE bioreactor™. Part I. Effect of the cell density on the process

High cell density perfusion process of antibody producing CHO cells was developed in disposable WAVE Bioreactor? using external hollow fiber filter as cell separation device. Both “classical” tangential flow filtration (TFF) and alternating tangential flow system (ATF) equipment were used and compared. Consistency of both TFF‐ and ATF‐based cultures was shown at 20–35 × 10⁶ cells/mL density stabilized by cell bleeds. To minimize the nutrients deprivation and by‐product accumulation, a perfusion rate correlated to the cell density was applied. The cells were maintained by cell bleeds at density 0.9–1.3 × 10⁸ cells/mL in growing state and at high viability for more than 2 weeks. Finally, with the present settings, maximal cell densities of 2.14 × 10⁸ cells/mL, achieved for the first time in a wave‐induced bioreactor, and 1.32 × 10⁸ cells/mL were reached using TFF and ATF systems, respectively. Using TFF, the cell density was limited by the membrane capacity for the encountered high viscosity and by the pCO₂ level. Using ATF, the cell density was limited by the vacuum capacity failing to pull the highly viscous fluid. Thus, the TFF system allowed reaching higher cell densities. The TFF inlet pressure was highly correlated to the viscosity leading to the development of a model of this pressure, which is a useful tool for hollow fiber design of TFF and ATF. At very high cell density, the viscosity introduced physical limitations. This led us to recommend cell densities under 1.46 × 10⁸ cell/mL based on the analysis of the theoretical distance between the cells for the present cell line. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:754–767, 2013     

Continuous rhamnolipid production using denitrifying Pseudomonas aeruginosa cells in hollow‐fiber bioreactor

Rhamnolipids are high‐value effective biosurfactants produced by Pseudomonas aeruginosa. Large‐scale production of rhamnolipids is still challenging especially under free‐cell aerobic conditions in which the highly foaming nature of the culture broth reduces the productivity of the process. Immobilized systems relying on oxygen as electron acceptor have been previously investigated but oxygen transfer limitation presents difficulties for continuous rhamnolipid production. A coupled system using immobilized cells and nitrate instead of oxygen as electron acceptor taking advantage of the ability of P. aeruginosa to perform nitrate respiration was evaluated. This denitrification‐based immobilized approach based on a hollow‐fiber setup eliminated the transfer limitation problems and was found suitable for continuous rhamnolipid production in a period longer than 1,500 h. It completely eliminated the foaming difficulties related to aerobic systems with a comparable specific productivity of 0.017 g/(g dry cells)‐h and allowed easy recovery of rhamnolipids from the cell‐free medium. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29: 346–351, 2013     

不同剂量丹参注射液联合泼尼松龙治疗口腔粘膜下纤维性变的临床研究

目的：探讨采用不同剂量的丹参注射液联合波尼松龙治疗口腔粘膜下纤维性病的治疗效果,为今后的治疗提供更多的依据。方法：选择从2010年1月至2013年1月期间在我院口腔科治疗的100例口腔粘膜下纤维性病患者,根据门诊号,随机将患者分为低剂量组、次低剂量组、中剂量组、高剂量组和对照组,每组各20例,低剂量组、次低剂量组、中剂量组、高剂量组,分别使用不同剂量丹参注射液联合波尼松龙治疗,对照组单纯使用波尼松龙治疗,观察治疗一个疗程后患者口腔粘膜情况及张口度。结果：中剂量组和高剂量组情况改善要明显好于低剂量、次低剂量组、对照组,差异具有显著性（P〈0．05）。结论：丹参注射液联合泼尼松龙治疗口腔粘膜下纤维性病疗效令人满意,其中低剂量丹参注射液便有效果,一定范围内剂量越高,疗效越好,值得在临床推f,     

A Fully Verified Theoretical Analysis of Contact‐Mode Triboelectric Nanogenerators as a Wearable Power Source

Harvesting mechanical energy from human activities by triboelectric nanogenerators (TENGs) is an effective approach for sustainable, maintenance‐free, and green power source for wireless, portable, and wearable electronics. A theoretical model for contact‐mode triboelectric nanogenerators based on the principles of charge conservation and zero loop‐voltage is illustrated. Explicit expressions for the output current, voltage, and power are presented for the TENGs with an external load of resistance. Experimental verification is conducted by using a laboratory‐fabricated contact‐mode TENG made from conducting fabric electrodes and polydimethylsiloxane/graphene oxide composite as the dielectric layer. Excellent agreements of the output voltage, current, and power are demonstrated between the theoretical and experimental results, without any adjustable parameters. The effects of the moving speed on output voltage, current, and power are illustrated in three cases, that is, the motion with constant speed, the sinusoidal motion cycles, and the real walking cycles by human subject. The fully verified theoretical model is a very powerful tool to guide the design of the device structure and selection of materials, and optimization of performance with respect to the application conditions of TENGs.