Extracellular Vesicles: Messengers of p53 in Tumor–Stroma Communication and Cancer Metastasis

Tumor progression to a metastatic and ultimately lethal stage relies on a tumor-supporting microenvironment that is generated by reciprocal communication between tumor and stromal host cells. The tumor–stroma crosstalk is instructed by the genetic alterations of the tumor cells—the most frequent being mutations in the gene Tumor protein p53 (TP53) that are clinically correlated with metastasis, drug resistance and poor patient survival. The crucial mediators of tumor–stroma communication are tumor-derived extracellular vesicles (EVs), in particular exosomes, which operate both locally within the primary tumor and in distant organs, at pre-metastatic niches as the future sites of metastasis. Here, we review how wild-type and mutant p53 proteins control the secretion, size, and especially the RNA and protein cargo of tumor-derived EVs. We highlight how EVs extend the cell-autonomous tumor suppressive activity of wild-type p53 into the tumor microenvironment (TME), and how mutant p53 proteins switch EVs into oncogenic messengers that reprogram tumor–host communication within the entire organism so as to promote metastatic tumor cell dissemination.     

聚合氯化铝投加时间对好氧颗粒污泥的形成和胞外聚合物的影响

通过在好氧颗粒污泥培养过程中分别在第1～7天和第8～14天投加聚合氯化铝(PAC)的方式,分析颗粒的形成过程,并研究胞外聚合物总量及其各组分的含量变化和空间分布。研究发现:第8～14天投加PAC的反应器形成颗粒时间提前了6天,且形成的颗粒外形规则,大小均匀,去除污染物性能良好;反应器中胞外多聚物和蛋白质含量总体较多,且随颗粒的形成而增加,之后略有下降并在颗粒成熟后维持稳定,蛋白质/多糖的值在颗粒形成期均增加超过2.5倍,有利于颗粒污泥的形成;多重荧光染色表明:两个反应器中PAC投加时间不同,β-D-呋喃葡萄糖、脂类和活细胞均呈现不同的分布情况,但对蛋白质和α-呋喃葡萄糖、α-甘露糖分布的影响较小。     

螺旋对称流厌氧反应器常温下的运行特性

螺旋对称流厌氧反应器（SSSAB）的最高有机负荷能在中温条件下（35℃）可达361.5kgCOD/（m³·d）,然而在常温条件下（10～30℃）其运行性能及污泥特征尚不明确,需进一步探究。以分段组合式厌氧反应器（TCAB）和升流式厌氧污泥床反应器（UASB）为参比,在常温下,以相同的操作条件研究了SSSAB的运行性能和污泥特征。实验结果表明：SSSAB的COD平均去除率（88%）高于TCAB和UASB（80%和78%）,SSSAB的一级反应动力学常数为5.4d^-1,高于TCAB的3.6d^-1和UASB的2.2d^-1。SSSAB内颗粒污泥宏观上相比于TCAB及UASB轮廓清晰、黑亮密实,且SSSAB颗粒污泥表面微观上较为粗糙,存在许多孔道。SSSAB颗粒污泥的胞外聚合物（EPS）总量高于TCAB及UASB,为底物的质量传递创造了条件,SSSAB颗粒污泥EPS的蛋白质（PN）/多糖（PS）较低,更有利于维持高污泥强度和优良的沉降性能。相比于UASB,SSSAB厌氧颗粒污泥凝聚性的分布更优,且其凝聚性波动更小。     

螺旋藻回用培养液组成对藻细胞生长的影响

将螺旋藻回用培养液补齐营养盐用于培养螺旋藻,考察了螺旋藻生物量、培养液中硝酸盐和磷酸盐及培养末期螺旋藻胞内生化成分含量对藻细胞生长的影响;用超滤膜对回用培养液中物质进行分子量分级,分析各级组分对螺旋藻的抑制效应,确定抑制物的分子量分布范围,并分析其成分. 结果表明,回用培养液中的螺旋藻胞外分泌物对藻细胞生长有明显抑制作用,比生长速率比新鲜培养基中低23%,且会抑制藻细胞对营养盐的吸收及胞内蛋白和叶绿素合成,对硝酸盐、磷酸盐的吸收分别低36%和37%,胞内蛋白及叶绿素含量分别为新鲜培养基中的0.85和0.65倍. 抑制物为分子量大于100 kDa的螺旋藻胞外多糖.     

Isoflurane Protects Against Human Endothelial Cell Apoptosis by Inducing Sphingosine Kinase-1 via ERK MAPK

Endothelial dysfunction is a major clinical problem affecting virtually every patient requiring critical care. Volatile anesthetics are frequently used during the perioperative period and protect the heart and kidney against ischemia and reperfusion injury. We aimed to determine whether isoflurane, the most commonly used volatile anesthetic in the USA, protects against endothelial apoptosis and necrosis and the mechanisms involved in this protection. Human endothelial EA.hy926 cells were pretreated with isoflurane or carrier gas (95% room air + 5% CO(2)) then subjected to apoptosis with tumor necrosis factor-α or to necrosis with hydrogen peroxide. DNA laddering and in situ Terminal Deoxynucleotidyl Transferase Biotin-dUTP Nick-End Labeling (TUNEL) staining determined EA.hy926 cell apoptosis and percent LDH released determined necrosis. We also determined whether isoflurane modulates the expression and activity of sphingosine kinase-1 (SK1) and induces the phosphorylation of extracellular signal regulated kinase (ERK MAPK) as both enzymes are known to protect against cell death. Isoflurane pretreatment significantly decreased apoptosis in EA.hy926 cells as evidenced by reduced TUNEL staining and DNA laddering without affecting necrosis. Mechanistically, isoflurane induces the phosphorylation of ERK MAPK and increased SK1 expression and activity in EA.hy926 cells. Finally, selective blockade of SK1 (with SKI-II) or S1P(1) receptor (with W146) abolished the anti-apoptotic effects of isoflurane. Taken together, we demonstrate that isoflurane, in addition to its potent analgesic and anesthetic properties, protects against endothelial apoptosis most likely via SK1 and ERK MAPK activation. Our findings have significant clinical implication for protection of endothelial cells during the perioperative period and patients requiring critical care.     

EPS对活性污泥絮凝沉降性能与表面性质的影响

采用序批式生物反应器（SBR）处理模拟废水,在葡萄糖作为碳源的条件下,调整进水碳源浓度改变系统内EPS含量,考察EPS的变化对活性污泥絮凝沉降性能及其表面性质的影响。结果表明,出水悬浮固体浓度（ESS）愈小,污泥絮体重絮能力（FA）越大,絮凝效果愈好。随着EPS的增加,ESS和污泥容积指数（SVI）升高,FA和ZSV降低,导致活性污泥絮凝性能和沉降性能下降。EPS与ESS呈正相关,EPS与FA呈负相关（R2分别为0.9916和-0.9941）,EPS与SVI呈正相关性,EPS与区域沉降速率（ZSV）呈负相关性（R2分别为0.9451和-0.9805）。同时,EPS的增加对污泥表面性质产生重要影响,EPS的增加导致污泥表面Zeta电位下降,污泥相对疏水性RH逐渐降低。Zeta电位的下降和RH的降低直接引起SVI和ESS增大,导致污泥絮凝沉降性能下降。EPS总量与Zeta电位和RH的相关性系数R2分别为-0.99、-0.9979。     

Tetrahydroisoquinolines: New Inhibitors of Neutrophil Extracellular Trap (NET) Formation

Neutrophils are short‐lived leukocytes that migrate to sites of infection as part of the acute immune response, where they phagocytose, degranulate, and form neutrophil extracellular traps (NETs). During NET formation, the nuclear lobules of neutrophils disappear and the chromatin expands and, accessorized with neutrophilic granule proteins, is expelled. NETs can be pathogenic in, for example, sepsis, cancer, and autoimmune and cardiovascular diseases. Therefore, the identification of inhibitors of NET formation is of great interest. Screening of a focused library of natural‐product‐inspired compounds by using a previously validated phenotypic NET assay identified a group of tetrahydroisoquinolines as new NET formation inhibitors. This compound class opens up new avenues for the study of cellular death through NET formation (NETosis) at different stages, and might inspire new medicinal chemistry programs aimed at NET‐dependent diseases.     

Vesicle-templating poly (3,4-ethylenedioxythiophene) hollow microspheres with enhanced electrocatalytic activity toward ascorbic acid oxidation

Poly(3,4-ethylenedioxythiophene) (PEDOT) hollow microspheres ranging from 50 to 950?nm are synthesized by chemically oxidative polymerization of 3,4-ethylenedioxythiophene using ammonium persulfate in the aqueous solution of cetyltrimethylammonium bromide (CTAB) and sodium dodecylbenzenesulfate (SDBS). Vesicles formed by CTAB and SDBS serve as templates for the formation of PEDOT hollow microspheres. The obtained PEDOT hollow microspheres were characterized by Fourier transform infrared spectroscopy, elemental analysis, X-ray photoelectron spectroscopy, and conductivity measurement. Compared to PEDOT granular particles, PEDOT hollow microspheres showed a more effective electrocatalytic activity in lowering the ascorbic acid oxidation potential.     

Gb3 Glycosphingolipids with Fluorescent Oligoene Fatty Acids: Synthesis and Phase Behavior in Model Membranes

Glycosphingolipids are involved in a number of physiological and pathophysiological processes, and they serve as receptors for a variety of bacterial toxins and viruses. To investigate their function in lipid membranes, fluorescently labeled glycosphingolipids are highly desirable. Herein, a synthetic route to access Gb₃ glycosphingolipids with fluorescently labeled fatty acids, consisting of pentaene and hexaene moieties either at the terminus or in the middle of the acyl chain, has been developed. The fluorescent properties of the Gb₃ derivatives were investigated in small unilamellar vesicles composed of a raft-like mixture. Phase-separated giant unilamellar vesicles (GUVs) allowed the quantification of the apparent partitioning coefficients of the Gb₃ compounds by means of confocal fluorescence laser scanning microscopy. The determined partition coefficients demonstrate that the Gb₃ derivatives are preferentially localized in the liquid-disordered (l_d) phase. To analyze whether the compounds behave like their physiological counterparts, Cy3-labeled (Cy: cyanine) Shiga toxin B subunits (STxB) were specifically bound to Gb₃-doped GUVs. However, the protein was favorably localized in the l_d phase, in contrast to results reported for STxB bound to naturally occurring Gb₃, which is discussed in terms of the packing density of the lipids in the liquid-ordered (l_o) phase.     

Imaging of Extracellular pH Using Hyperpolarized Molecules

Many diseases can overrule natural pH regulatory mechanisms and alter the extracellular pH (pH_e). A non-invasive method that resolves pH_e in vivo with high spatial and temporal resolution could therefore improve diagnosis and monitoring of diseases, contributing to the concept of precision medicine. During the last decades, several techniques have been proposed to image pH_e non-invasively. The majority of these methods rely on magnetic resonance because of its good spatial resolution, high penetration depth, non-ionizing radiation and excellent complimentary soft tissue contrast. Dissolution dynamic nuclear polarization (DNP) is an emerging concept to enhance nuclear magnetic resonance (NMR) signals by more than four orders of magnitude, making it possible to observe in vivo metabolic processes in real-time. Here, we summarize and review recent developments in pH_e imaging techniques based on hyperpolarization methods and give an overview of recently discovered hyperpolarized pH sensor molecules that have been applied in vitro and in vivo.