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.     

Formation of phase structure and crystallization behavior in blends containing polystyrene–polyethylene block copolymers

The microphase separation structure in the molten state and the structure formation in crystallization from such ordered melt were investigated for the blends of polystyrene–polyethylene block copolymers (SE) with polystyrene homopolymer (PS) and polyethylene homopolymer (PE) and for the blends consisting of two kinds of SE with different copolymer compositions from each other, using synchrotron small-angle X-ray scattering techniques (SAXS). The copolymer compositions of SE block copolymers employed were 0.34, 0.58 and 0.73 wt. fraction of PE, and their melt morphologies were cylindrical, lamellar and lamellar, respectively. Macrophase separation or the morphology change in the melt occurred depending on the molecular weight and the blend composition, as reported so far. In crystallization from such macrophase-separated and microphase-separated melts, the melt morphology was completely kept for all the blends. Crystallization behavior was also investigated for the blends. The crystallization within the spherical and cylindrical domains surrounded by glassy PS was not observed for SE/PS blends. In the crystallization from the macrophase-separated melt, two exothermal peaks were observed in the DSC measurements, while a single peak was observed for other blends. For the blends with PS, the degree of crystallinity was depressed and the apparent activation energy of crystallization was high, compared to those for the corresponding neat SE. For SE/PE and SE/SE blends, those were changed depending on the blend composition.     

Evolution of patinas on copper exposed in a suburban area

Copper plates were exposed under sheltered outdoor conditions for up to one year, starting in September 2001 in Musashino City, Tokyo, a suburban area. Following various periods of exposure, the patinas on the plates were characterized to investigate their evolution by using X-ray fluorescence analysis, X-ray diffraction, field emission scanning electron microscopy, and glow discharge optical emission spectroscopy. The difference in the roles of sulfur and chlorine in the early stages of copper patination were identified by analyzing the depth profiles of these two elements. Sulfur was found on top of the patina as cupric sulfates such as posnjakite (Cu₄SO₄(OH)₆ · H₂O) or brochantite (Cu₄SO₄(OH)₆). Brochantite appeared only after 12 months of exposure. In contrast, chlorine was found on the surface after only one month of exposure. It gradually penetrated the patina as the exposure period lengthened, forming copper chloride complexes. Chloride ions accumulated at the patina/copper interface, forming nantokite (CuCl), which promoted corrosion.     

Differences between corrosion products formed on copper exposed in Tokyo in summer and winter

We analyzed the copper corrosion products that formed during a month in summer and a month in winter at three sites in Tokyo using several analytical techniques. The X-ray diffraction patterns revealed that cuprite Cu₂O and posnjakite Cu₄SO₄(OH)₆·H₂O formed on copper exposed in summer. By contrast, only cuprite was found in winter exposed copper. The X-ray fluorescence results indicated that the amounts of sulfur and chlorine on the copper plates exposed in summer were much greater than those in winter. This could be explained by the change in particulate sulfate and sea salt concentrations. Depth profiling analysis by Auger electron spectroscopy revealed that the oxide layer formed in summer was thicker than that in winter. This difference in oxide layer thickness could have been due to the differences in temperature, relative humidity, and the amount of sulfur and chlorine on the copper plate.     

Sequential transformation and utilization of natural network polymer “LIGNIN”

Lignin is the most abundant and complicated natural polymer, forming long-term carbon flow in the ecosystem. Lignin is composed of phenyl propane units which are the combination of glycerol- and polyhydric phenol units. A key for sequential functionality control is how to control C1- (7%) and C2- (50%) aryl ethers selectively. The cleavage of C1 aryl ethers results in the transformation of polymer structures from network to linear-type, and that at C2 leads to depolymerization to dimer–oligomer level. The originally designed sequential transformation system includes two steps: 1st step; the selective control of C1 structures to give 1,1-bis(aryl)propane type units, leading to the standardization of C1 functionality, 2nd step; the selective control of C2 structures using intramolecular switching devices [1,1-bis(aryl)propane-2-O-aryl ether units], leading to the change of molecular weight and phenolic activity. The polymers, lignophenols, derived from native lignin through C1 control are multi-functional lignin-based materials, linear types of lignin subunits with phenol derivatives at C1. Sequential applications have been designed originally.     

Bacteriophages isolated from activated sludge processes and their polyvalency

In this study, bacteriophages were isolated from activated sludge and their host range was studied. Bacterial isolates were obtained from an activated sludge process treating urban sewage, and bacteriophages were obtained by plaque assay using the bacterial isolates obtained in this study as the host. Out of 15 bacteria isolated, 9 supported plaque formation. The host range test was conducted with a combination of 8 bacteriophage isolates and 9 bacterial isolates. All of the 8 bacteriophages tested were found to form plaques on more than 1 host, and 4 of them formed plaques on both gram-positive and gram-negative bacterial isolates. Three of the 8 bacteriophages failed to form plaques on their original bacterial host. The experimental result indicates that bacteriophages are an active part of the activated sludge microbial ecosystem, having a very close ecological relationship with their host bacteria.     

Space charge behavior in polymer film based on increment of capacitance under AC high field

Polymer materials have excellent dielectric and insulation properties; however, those properties in AC high field region have not been known well. Recently we established an evaluation method of high‐field AC dissipation current waveform of polymer materials 1 . AC dissipation current waveforms of polyethylene and polypropylene films show nonlinear distortion in AC high‐field region. This nonlinearity was thought to be related to the behavior of AC space charge formation in the sample near electrodes. The properties of space charge formed under AC high field at power frequency seem to differ from those formed under DC high field. The measurement of AC space charge distribution is not so easy due to the resolution limit of the space charge measurement. We studied the dielectric properties of biaxially oriented polypropylene (BOPP) film under AC high field up to 120 °C. It was found that tan δ, AC dissipation current (Ixr), and unbalanced component of capacitive current (ΔIxc) increased when the temperature became higher. In particular, ΔIxc increased above some threshold field and was considered to be due to the AC space charge formation. This AC space charge layer near electrode is thought to be formed due to carrier injection under AC high‐field application. Usually, the carrier mobility becomes smaller on lowering the temperature. Most of the carriers injected from the electrode are trapped near the electrode in the sample film. But in the high‐temperature region, the carrier mobility becomes larger and the carrier injection starts to increase from lower field. Many more carriers are injected from the electrode. It is thought that some of the injected carriers are trapped inside the sample film; the others go through the sample to the opposite side. © 2002 Wiley Periodicals, Inc. Electr Eng Jpn, 141(2): 8–16, 2002; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/eej.10018