Visualizing Extracellular Vesicles and Their Function in 3D Tumor Microenvironment Models

Extracellular vesicles (EVs) are cell-derived nanostructures that mediate intercellular communication by delivering complex signals in normal tissues and cancer. The cellular coordination required for tumor development and maintenance is mediated, in part, through EV transport of molecular cargo to resident and distant cells. Most studies on EV-mediated signaling have been performed in two-dimensional (2D) monolayer cell cultures, largely because of their simplicity and high-throughput screening capacity. Three-dimensional (3D) cell cultures can be used to study cell-to-cell and cell-to-matrix interactions, enabling the study of EV-mediated cellular communication. 3D cultures may best model the role of EVs in formation of the tumor microenvironment (TME) and cancer cell-stromal interactions that sustain tumor growth. In this review, we discuss EV biology in 3D culture correlates of the TME. This includes EV communication between cell types of the TME, differences in EV biogenesis and signaling associated with differing scaffold choices and in scaffold-free 3D cultures and cultivation of the premetastatic niche. An understanding of EV biogenesis and signaling within a 3D TME will improve culture correlates of oncogenesis, enable molecular control of the TME and aid development of drug delivery tools based on EV-mediated signaling.     

Simulated Microgravity Inhibits the Proliferation of Chang Liver Cells by Attenuation of the Major Cell Cycle Regulators and Cytoskeletal Proteins

Simulated microgravity (SMG) induced the changes in cell proliferation and cytoskeleton organization, which plays an important factor in various cellular processes. The inhibition in cell cycle progression has been considered to be one of the main causes of proliferation inhibition in cells under SMG, but their mechanisms are still not fully understood. This study aimed to evaluate the effects of SMG on the proliferative ability and cytoskeleton changes of Chang Liver Cells (CCL-13). CCL-13 cells were induced SMG by 3D clinostat for 72 h, while the control group were treated in normal gravity at the same time. The results showed that SMG reduced CCL-13 cell proliferation by an increase in the number of CCL-13 cells in G0/G1 phase. This cell cycle phase arrest of CCL-13 cells was due to a downregulation of cell cycle-related proteins, such as cyclin A1 and A2, cyclin D1, and cyclin-dependent kinase 6 (Cdk6). SMG-exposed CCL-13 cells also exhibited a downregulation of α-tubulin 3 and β-actin which induced the cytoskeleton reorganization. These results suggested that the inhibited proliferation of SMG-exposed CCL-13 cells could be associate with the attenuation of major cell cycle regulators and main cytoskeletal proteins.     

Clinical Significance of Tumor Infiltrating Lymphocytes in Association with Hormone Receptor Expression Patterns in Epithelial Ovarian Cancer

Hormone receptor expression patterns often correlate with infiltration of specific lymphocytes in tumors. Specifically, the presence of specific tumor-infiltrating lymphocytes (TILs) with particular hormone receptor expression is reportedly associated with breast cancer, however, this has not been revealed in epithelial ovarian cancer (EOC). Therefore, we investigated the association between hormone receptor expression and TILs in EOC. Here we found that ERα, AR, and GR expression increased in EOC, while PR was significantly reduced and ERβ expression showed a reduced trend compared to normal epithelium. Cluster analysis indicated poor disease-free survival (DFS) in AR+/GR+/PR+ subgroup (triple dominant group); while the Cox proportional-hazards model highlighted the triple dominant group as an independent prognostic factor for DFS. In addition, significant upregulation of FoxP3+ TILs, PD-1, and PD-L1 was observed in the triple dominant group compared to other groups. NanoString analyses further suggested that tumor necrosis factor (TNF) and/or NF-κB signaling pathways were activated with significant upregulation of RELA, MAP3K5, TNFAIP3, BCL2L1, RIPK1, TRAF2, PARP1, and AKT1 in the triple dominant EOC group. The triple dominant subgroup correlates with poor prognosis in EOC. Moreover, the TNF and/or NF-κB signaling pathways may be responsible for hormone-mediated inhibition of the immune microenvironment.     

TNFα-Induced LDL Cholesterol Accumulation Involve Elevated LDLR Cell Surface Levels and SR-B1 Downregulation in Human Arterial Endothelial Cells

Excess lipid droplets are frequently observed in arterial endothelial cells at sites of advanced atherosclerotic plaques. Here, the role of tumor necrosis factor alpha (TNFα) in modulating the low-density lipoprotein (LDL) content in confluent primary human aortic endothelial cells (pHAECs) was investigated. TNFα promoted an up to 2 folds increase in cellular cholesterol, which was resistant to ACAT inhibition. The cholesterol increase was associated with increased ¹²⁵I-LDL surface binding. Using the non-hydrolysable label, Dil, TNFα could induce a massive increase in Dil-LDL by over 200 folds. The elevated intracellular Dil-LDL was blocked with excess unlabeled LDL and PCSK9, but not oxidized LDL (oxLDL), or apolipoprotein (apoE) depletion. Moreover, the TNFα-induced increase of LDL-derived lipids was elevated through lysosome inhibition. Using specific LDLR antibody, the Dil-LDL accumulation was reduced by over 99%. The effects of TNFα included an LDLR cell surface increase of 138%, and very large increases in ICAM-1 total and surface proteins, respectively. In contrast, that of scavenger receptor B1 (SR-B1) was reduced. Additionally, LDLR antibody bound rapidly in TNFα-treated cells by about 30 folds, inducing a migrating shift in the LDLR protein. The effect of TNFα on Dil-LDL accumulation was inhibited by the antioxidant tetramethythiourea (TMTU) dose-dependently, but not by inhibitors against NF-κB, stress kinases, ASK1, JNK, p38, or apoptosis caspases. Grown on Transwell inserts, TNFα did not enhance apical to basolateral LDL cholesterol or Dil release. It is concluded that TNFα promotes LDLR functions through combined increase at the cell surface and SR-B1 downregulation.     

Embryonic Trophectoderm Secretomics Reveals Chemotactic Migration and Intercellular Communication of Endometrial and Circulating MSCs in Embryonic Implantation

Embryonic implantation is a key step in the establishment of pregnancy. In the present work, we have carried out an in-depth proteomic analysis of the secretome (extracellular vesicles and soluble proteins) of two bovine blastocysts embryonic trophectoderm primary cultures (BBT), confirming different epithelial–mesenchymal transition stages in these cells. BBT-secretomes contain early pregnancy-related proteins and angiogenic proteins both as cargo in EVs and the soluble fraction. We have demonstrated the functional transfer of protein-containing secretome between embryonic trophectoderm and maternal MSC in vitro using two BBT primary cultures eight endometrial MSC (eMSC) and five peripheral blood MSC (pbMSC) lines. We observed that eMSC and pbMSC chemotax to both the soluble fraction and EVs of the BBT secretome. In addition, in a complementary direction, we found that the pattern of expression of implantation proteins in BBT-EVs changes depending on: (i) their epithelial–mesenchymal phenotype; (ii) as a result of the uptake of eMSC- or pbMSC-EV previously stimulated or not with embryonic signals (IFN-τ); (iii) because of the stimulation with the endometrial cytokines present in the uterine fluid in the peri-implantation period.     

纳米晶体Se的微观结构特征

本文采用Ｘ射线衍射和透射电子显微镜研究了纳米晶体Ｓｅ的微观结构特征．通过测定不同晶粒尺寸的纳米晶体Ｓｅ的点阵参数ａ和ｃ，发现纳米晶体Ｓｅ中存在晶格畸变，畸变量与晶粒尺寸有关．得到了纳米晶体Ｓｅ中晶胞体积变化与晶粒尺寸的关系．并由点阵参数计算出纳米晶体Ｓｅ的键长．用非晶态Ｓｅ中的无规链折叠的晶化机制解释了纳米晶体Ｓｅ中晶格畸变现象．     

高技术产品用无源元件与中国钽工业发展

世界高技术产品市场总体看呈增长趋势，预计2020年销售额将比2000年翻一番。以高技术产品为主要应用领域的电阻、电容、电感等无源元件，也将随着高技术产业的发展呈现出良好的市场前景，但是钽电容器市场近几年表现出骤涨急落的发展态势。由于钽电容器产能过剩，且面临着被替代的威协，因此价格将无法返回到2000年以前的水平，并呈现整体持续走低的趋势。中国的钽工业发展应注重提高企业自身的综合竞争力，加强企业间的沟通、交流与合作，维护我国钽工业的整体利益，增强环保意识，做好国外转移到我国的初级产品和低端产品生产的环保工作。     

An experimental evaluation of the effects of ripple current generated by the power conditioning stage on a proton exchange membrane fuel cell stack

In this article, an impedance model of the proton exchange membrane fuel cell stack (PEMFCS) is proposed. The proposed study employs an equivalent circuit of the PEMFCS derived by the frequency response analysis technique. An equivalent circuit for the PEMFCS is developed to evaluate the effects of ripple currents generated by the power-conditioning unit. The calculated results are then verified by means of experiments using a commercially available PEMFCS. The relationship between ripple current and fuel cell performance, such as power loss and fuel consumption, is investigated. Experimental results show that the ripple current can contribute up to a 6% reduction in the available output power. This paper was presented at the Fuel Cells: Materials, Processing, and Manufacturing Technologies Symposium sponsored by the Energy/Utilities Industrial Sector & Ground Transportation Industrial Sector and the Specialty Materials Critical Technologies Sector at the ASM International Materials Solutions Conference, October 13–15, 2003, in Pittsburgh, PA. The symposium was organized by P. Singh, Pacific Northwest National Laboratory, S.C. Deevi, Philip Morris USA, T. Armstrong, Oak Ridge National Laboratory, and T. Dubois, U.S. Army CECOM.     

氯化铜在水溶液中的扩散系数的测定SCIEI

用膈膜池法测定了298K条件下CuCl_2在CuCl_2和CuCl_2-HCl水溶液中的扩散系数。发现CuCl_2的扩散系数与其浓度依赖性主要取决于它的化学位和溶液的粘度。     

Effect of oxygen precipitates in solar grade silicon on minority carrier lifetime and efficiency of solar cells

The effect of oxygen precipitates on minority carrier lifetime and performance of solar cell was studied by means of Fourier Transform Infrared Spectroscopy (FTIR), quasi-steady state photoconductance (QSSPCD), optical microscope, spectrumresponse and solar cell efficiency test. The minority carrier lifetime and performance of solar cell reduced depend on oxygen precipitates. A few of oxygen precipitates have formed after single-step annealing; and they do not impact the efficiency dramatically. Pre-annealing at 650 ℃ for 4 h enhances the oxygen precipitation when it is subjected to middle temperature annealing. The solar cells performance decayed sharply. Especially annealing at 950 ℃ for 3 h, the V os and I sc of cells decrease 12% and 25% respectively. Few oxygen precipitates have formed in silicon after high temperature annealing at about 1050 ℃ whether pre-annealing is used or not, and the performance of cells is notbe affected.