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.     

Rapid consolidation of powdered materials by induction hot pressing

A rapid hot press system in which the heat is supplied by RF induction to rapidly consolidate thermoelectric materials is described. Use of RF induction heating enables rapid heating and consolidation of powdered materials over a wide temperature range. Such rapid consolidation in nanomaterials is typically performed by spark plasma sintering (SPS) which can be much more expensive. Details of the system design, instrumentation, and performance using a thermoelectric material as an example are reported. The Seebeck coefficient, electrical resistivity, and thermal diffusivity of thermoelectric PbTe material pressed at an optimized temperature and time in this system are shown to agree with material consolidated under typical consolidation parameters.     

Development of a homogeneous competitive immunoassay for phosphorylated protein antigen based on the enhanced fluorescence resonance energy transfer technology

We describe a homogeneous competitive immunoassay for a phosphorylated protein antigen. The assay takes advantage of the enhanced fluorescence resonance energy transfer (FRET) technology, which has a unique characteristic that the FRET signal is increased by the specific interaction of two fluorolabeled leucine zippers. We chose extracellular signal-regulated kinase (ERK) as a model antigen and constructed two molecular probes in which either anti-phosphorylation site antibody or the antigen peptide was chemically conjugated to the enhanced FRET probes. While these molecular probes indicated sufficient FRET signal without antigen, they displayed a significant change in the fluorescent spectrum by mixing with phosphorylated antigens. With this competitive enhanced FRET immunoassay, a phosphorylated ERK concentration within the range from 15 nM to 250 nM could be determined. Because the assay is very simple, it would be applied to not only in vitro assay but also in vivo detection of protein phosphorylation.     

The development of pivot bearing with multiple degrees of freedom: 1st report: Prototype of pivot bearing with two degrees of freedom

This paper deals with the prototype of a new pivot bearing having two degrees of freedom. The idea of the pivot bearing is based on a continuous velocity joint (CVJ). The experimental axial stiffness and contact pressure are compared with those determined by theoretical analysis. Then, it is confirmed that the pivot bearing swings smoothly with a range of ±25°. Furthermore, the stiffness of the bearing increases as the swinging angle becomes larger. Therefore, this newly developed pivot bearing may be applied to a parallel mechanism, a joint of robot and so on.     

Application of electron beam for the reduction of PCDD/F emission from municipal solid waste incinerators

The electron-beam technology was applied to reduce the emission of polychlorinated dibenzo-p-dioxins (PCDD) and polychlorinated dibenzofurans (PCDF) in a flue gas of 1000 m(3)N/h from the municipal solid waste incinerator (MSWI) at a temperature of 200 degrees C. More than 90% decomposition of PCDD/Fs was obtained using an electron accelerator at a dose of 14 kGy. The decomposition was initiated through reactions with OH radicals produced by the irradiation of flue gases, followed by oxidation such as the ring cleavage of the aromatic ring, the dissociation of ether bond, and dechlorination. The cost analysis estimated that the electron-beam system can cut the annualized cost by approximately 50% for the treatment of PCDD/Fs in a pre-dusted MSWI flue gas as compared with a bag-filter system when operating on electricity generated from an incineration. Electron-beam technology is an economically and technologically useful method for reducing PCDD/Fs in an incineration flue gas.     

Determination of the solar calibration constant for a sun-sky radiometer: proposal of an in-situ procedure

A new procedure is presented for determining in situ the solar calibration constant, i.e., the Sun-sky radiometer counts for a direct normal solar flux extrapolated to the top of the atmosphere. The method makes use of a modified version of the Langley plot based on the use of an inversion code of column-integrated aerosol size distribution, and it is ordinarily applied to calibrate Prede Sun-sky radiometers. To analyze how such an in situ method can work accurately, the technique has been applied to a five-month dataset obtained from measurements taken in Rome, Italy, by a Prede Sun-sky radiometer from 22 April to 5 November 2001. The precision of the in situ method has been estimated to within 1-2.5%, depending on the wavelength.     

Rule-Based Building-Block Architectures for Policy-Based Networking

We developed two rule-based building-block architectures, i.e., pipe-connection and label-connection architectures, for describing complex and structured policies, especially network QoS policies. This study focuses on the latter. The relationships or connections between building blocks are specified by the data flow and control flow between them. The data flow is specified by tags, including virtual flow labels (VFLs), which are data attached to outside packets. The control flow can be classified and specified by four control structures: concatenation, parallel application, selection, and repetition. We have designed fine-grained and coarse-grained building blocks and methods for specifying data flow and control flow in differentiated services (Diffserv), and implemented the coarse-grained ones in a policy server. Two cases of building-block use are described, and we concluded that there are five advantages of building-block-based policies, i.e., expressibility, uniform semantics, simplicity, flexibility, and management-task-oriented design. We also developed techniques for transforming building-block policies into executable ones, which are called policy division and fusion.     

Hydrothermal synthesis of surface-modified iron oxide nanoparticles

We synthesized surface-modified iron oxide nanoparticles in aqueous phase by heating an aqueous solution of iron sulfate (FeSO₄) at 473 K with a small amount of either n-decanoic acid (C₉H₁₉COOH) or n-decylamine (C₁₀H₂₁NH₂), which is not miscible with water at room temperature. Transmission electron microscopy showed that the addition of n-decanoic acid or decylamine changed the shape of the obtained nanoparticles. X-ray diffraction spectra revealed that the synthesized nanoparticles were in α-Fe₂O₃ or Fe₃O₄ phase while Fourier transform infrared spectroscopy and thermogravimetry indicated the existence of an organic layer on the surface of the nanoparticles. In the synthetic condition, decreased dielectric constant of water at higher temperature increased the solubility of n-decanoic acid or n-decylamine in water to promote the reaction between the surface of iron oxide nanoparticles and the organic reagents. After the synthesis, the used organic modifiers separated from the aqueous phase at room temperature, which may help the environmentally benign synthesis of surface-modified metal oxide nanoparticles.     

Genetically engineered active Qbeta replicase in rabbit reticulocyte cell-free system: a fusion protein of EF-Tu and EF-Ts is functional as the subunit of Qbeta replicase

Qbeta replicase functioning in Escherichia coli is an RNA-dependent RNA polymerase composed of one phage-coded subunit and three host-coded proteins: ribosomal protein S1, and protein elongation factors EF-Tu and EF-Ts. Qbeta replicase lacking ribosomal protein S1 (alpha-less replicase) is capable of replicating some small RNAs. We attempted to create functional alpha-less replicase by co-expression of the mRNAs that code for the subunits of alpha-less replicase in a rabbit reticulocyte cell-free translation system. Replicase activity, however, could not be detected when both EF-Tu and EF-Ts were co-expressed with the phage-coded subunit. On the other hand, active alpha-less replicase was obtained when an EF-Ts-EF-Tu fusion protein was co-expressed with the phage-coded subunit. Consequently, we succeeded in generating genetically engineered active alpha-less Qbeta replicase which functions in a eukaryotic cell-free system.     

Technology initiatives for the near future in internetworking

The introduction of the Open Computer Network (OCN), providing Internet-like services by NTT, will lead to higher-level demands by users. For example, users will be asking for higher quality, greater functionality, and lower charges for high-speed, high-throughput data transfer. To respond effectively to the wishes of sophisticated and varied users, technology development must be pursued from the user's point of view. For this reason, research and development is being performed on middleware to open up network functions and make it possible for the user and network to cooperate with each other. This article discusses the development of major technologies for achieving an enhanced OCN (next-stage OCN) which should become a catalyst far the future multimedia network, and presents a future vision for the network