Status of the Electrostatic Levitation Furnace (ELF) in the ISS-KIBO

The electrostatic levitation method is a containerless processing technique that utilizes Coulomb force between a charged sample and the surrounding electrodes. The Japan Aerospace Exploration Agency (JAXA) has been developing this technique for more than 20 years. In 2016, JAXA completed the flight model assembly, and the Electrostatic Levitation Furnace (ELF) for the International Space Station (ISS) was launched to the ISS. The ELF is mainly intended to handle oxide melts that are difficult to levitate on the ground based electrostatic levitator due to gravity and due to insufficient charging. ISS-ELF can measure the thermophysical properties (density, surface tension and viscosity) of high temperature melts above 2000 ^°C. The thermophysical properties data of materials at high temperature is useful for the study of liquid states and improvement of numerical simulation by modeling the manufacturing processes using the liquid state. Moreover, the interfacial energy of immiscible melts will be measured by creating a core-shell droplet configuration which otherwise cannot be obtained on the ground due to sedimentation. This paper briefly describes the ELF facility and presents the results of a functional checkout that includes the density measurement of molten alumina.     

Immunotherapy with 4-1BBL-Expressing iPS Cell‐Derived Myeloid Lines Amplifies Antigen-Specific T Cell Infiltration in Advanced Melanoma

We have established an immune cell therapy with immortalized induced pluripotent stem-cell–derived myeloid lines (iPS-ML). The benefits of using iPS-ML are the infinite proliferative capacity and ease of genetic modification. In this study, we introduced 4-1BBL gene to iPS-ML (iPS-ML-41BBL). The analysis of the cell-surface molecules showed that the expression of CD86 was upregulated in iPS-ML-41BBL more than that in control iPS-ML. Cytokine array analysis was performed using supernatants of the spleen cells that were cocultured with iPS-ML or iPS-ML-41BBL. Multiple cytokines that are beneficial to cancer immunotherapy were upregulated. Peritoneal injections of iPS-ML-41BBL inhibited tumor growth of peritoneally disseminated mouse melanoma and prolonged survival of mice compared to that of iPS-ML. Furthermore, the numbers of antigen-specific CD8⁺ T cells were significantly increased in the spleen and tumor tissues treated with epitope peptide-pulsed iPS-ML-41BBL compared to those treated with control iPS-ML. The number of CXCR6-positive T cells were increased in the tumor tissues after treatment with iPS-ML-41BBL compared to that with control iPS-ML. These results suggest that iPS-ML-41BBL could activate antigen-specific T cells and promote their infiltration into the tumor tissues. Thus, iPS-ML-41BBL may be a candidate for future immune cell therapy aiming to change immunological “cold tumor” to “hot tumor”.     

Anti-Oncogenic gem-Dihydroperoxides Induce Apoptosis in Cancer Cells by Trapping Reactive Oxygen Species

Organic gem-dihydroperoxides (DHPs) and their derived peroxides have attracted a great deal of attention as potential anti-cancer agents. However, the precise mechanism of their inhibitory effect on tumors is unknown. To determine the mechanism of the inhibitory effects of DHPs, we examined the effects of DHPs on leukemia K562 cells. As a result, certain DHPs used in this study exhibited growth-inhibitory activity according to a clear structure-activity relationship. The most potent DHP, 12AC3O, induced apoptosis in K562 cells, but not in peripheral blood monocytes (PBMCs) or fibroblast cells. 12AC3O induced apoptosis through the intrinsic mitochondrial pathway and thereafter through the extrinsic pathway. The activity of the former pathway was partly attenuated by a JNK inhibitor. Interestingly, 12AC3O induced apoptosis by trapping a large amount of ROS, leading to an extremely lower intracellular ROS level compared with that in the cells in the steady-state condition. These results suggest that an appropriate level of intracellular ROS was necessary for the maintenance of cancer cell growth. DHPs may have a potential to be a novel anti-cancer agent with minimum adverse effects on normal cells.     

Evaluation of crack nucleation site and mechanical properties for friction stir welded butt joint in 2024-T3 aluminum alloy

In this paper, metallographic observations, hardness measurement, and static and fatigue tests were conducted to investigate the discontinuity states which become crack nucleation sites in friction stir welded butt joints in 2-mm-thick 2024-T3 aluminum alloy and static and fatigue properties of the joint. Because different types of surface finish can be used depending on the application of the joint, several types of surface conditions were tested to evaluate their effect on crack nucleation sites and static and fatigue life. Indentation hardness tests revealed that typical hardness reduction is not necessarily observed on the section of the welding line. Based on fatigue test results, it was confirmed that there are several types of crack nucleation sites for friction stir welding (FSW) joints depending on the surface finish, and the features of the fracture surface also differ depending on the site. Furthermore, the type of discontinuity state affects the fatigue life of the FSW joint.     

Free Radical Scavenging and Cellular Antioxidant Properties of Astaxanthin

Astaxanthin is a coloring agent which is used as a feed additive in aquaculture nutrition. Recently, potential health benefits of astaxanthin have been discussed which may be partly related to its free radical scavenging and antioxidant properties. Our electron spin resonance (ESR) and spin trapping data suggest that synthetic astaxanthin is a potent free radical scavenger in terms of diphenylpicryl-hydrazyl (DPPH) and galvinoxyl free radicals. Furthermore, astaxanthin dose-dependently quenched singlet oxygen as determined by photon counting. In addition to free radical scavenging and singlet oxygen quenching properties, astaxanthin induced the antioxidant enzyme paroxoanase-1, enhanced glutathione concentrations and prevented lipid peroxidation in cultured hepatocytes. Present results suggest that, beyond its coloring properties, synthetic astaxanthin exhibits free radical scavenging, singlet oxygen quenching, and antioxidant activities which could probably positively affect animal and human health.     

Extracellular Disposal of Tumor-Suppressor miRs-145 and -34a via Microvesicles and 5-FU Resistance of Human Colon Cancer Cells

The dysregulation of microRNA (miRNA) expression causes various kinds of diseases. Especially, alterations in miRNA expression levels are frequently observed in human tumor cells and are associated with cancer pathogenesis. Earlier we established Fluorouracil (5-FU)-resistant human colon cancer DLD-1 cells (DLD-1/5FU) from parental 5-FU- sensitive DLD-1 cells. In the present study, we examined the expression of miRNA in each cell line and in its extracellular microvesicles (MVs) before and after treatment with 5-FU. The nascent RNAs of anti-oncogenic miR-34a and -145 labeled with EU in both cells were proved to be transferred into MVs in both cell lines. The levels of miR-34a and -145 in the cells and in their MVs were not largely different in the two cell lines, and a substantial amount of both miRNAs was secreted by both cell lines even in the steady-state condition. The exposure of both cell lines to 5-FU significantly increased the intracellular levels of miR-145 and miR-34a in the 5-FU-sensitive DLD-1 cells, whereas the level of neither miR was elevated in the DLD-1/5FU cells. Interestingly, the amount of miR-145 detected in the small MVs shed into the medium of the parental cells was reduced after the treatment with 5-FU. On the other hand, the intracellular expression of miR-34a in the DLD-1/5FU cells was down-regulated compared with that in the parental DLD-1 cells even in the steady-state condition. As to the miR-34a secreted into MVs, the increase in the level in DLD-1/5FU cells was greater than that in the parental DLD-1 cells after the treatment with 5-FU. Thus, the intra- and extracellular miR-145 and -34a were closely associated with 5-FU resistance, and the resistance was in part due to the enhanced secretion of miR-145 and -34a via MVs, resulting in low intracellular levels of both miRNAs.     

Discrimination of cucumber cultivars using a multiple‐point sheet sensor to measure biting force

To define the textural differences among three cucumber cultivars, we combined two different mechanical measurements: a direct measurement of mastication force using a multiple‐point sheet sensor (MSS), and a uniaxial compression test. For mastication force measurement, subjects bit a sample placed on the sheet sensor with their upper and lower incisors. During a single bite, masticatory force was measured using a computerised MSS device called the ISCAN system with a TSCAN sheet sensor. Values of maximum bite force, the time at which maximum bite force appeared, area under the force–time curve, contact area and maximum pressure point for one cultivar were significantly higher than those for the other two. For all the parameters, lower values corresponded to crispness and higher values to toughness. Overall tendencies were the same in the compression test, though the compression test tends to yield smaller standard deviations. The MSS masticatory test is suitable for analysing masticatory properties with respect to each individual, while the instrument test is suitable for analysing physical properties inherent in each sample. Combination of the actual mastication test performed in the human oral cavity and conventional mechanical tests provides the most reliable objective analysis of food texture. Copyright © 2003 Society of Chemical Industry     

Effect of introduction of chondroitin sulfate into polymer-peptide conjugate responding to intracellular signals

We recently developed a novel tumor-targeted gene delivery system responding to hyperactivated intracellular signals. Polymeric carrier for gene delivery consists of hydrophilic neutral polymer as main chains and cationic peptide substrate for target enzyme as side chains, and was named polymer-peptide conjugate (PPC). Introduction of chondroitin sulfate (CS), which induces receptor-medicated endocytosis, into polymers mainly with a high cationic charge density such as polyethylenimine can increase tumor-targeted gene delivery. In the present study, we examined whether introduction of CS into PPC containing five cationic amino acids can increase gene expression in tumor cells. Size and zeta potential of plasmid DNA (pDNA)/PPC/CS complex were <200 nm and between -10 and -15 mV, respectively. In tumor cell experiments, pDNA/PPC/CS complex showed lower stability and gene regulation, compared with that of pDNA/PPC. Moreover, no difference in gene expression was identified between positive and negative polymer. These results were caused by fast disintegration of pDNA/PPC/CS complexes in the presence of serum. Thus, we suggest that introduction of negatively charged CS into polymers with a low charge density may lead to low stability and gene regulation of complexes.