Metastatic Voyage of Ovarian Cancer Cells in Ascites with the Assistance of Various Cellular Components

Epithelial ovarian cancer (EOC) is the most lethal gynecologic malignancy and has a unique metastatic route using ascites, known as the transcoelomic root. However, studies on ascites and contained cellular components have not yet been sufficiently clarified. In this review, we focus on the significance of accumulating ascites, contained EOC cells in the form of spheroids, and interaction with non-malignant host cells. To become resistant against anoikis, EOC cells form spheroids in ascites, where epithelial-to-mesenchymal transition stimulated by transforming growth factor-β can be a key pathway. As spheroids form, EOC cells are also gaining the ability to attach and invade the peritoneum to induce intraperitoneal metastasis, as well as resistance to conventional chemotherapy. Recently, accumulating evidence suggests that EOC spheroids in ascites are composed of not only cancer cells, but also non-malignant cells existing with higher abundance than EOC cells in ascites, including macrophages, mesothelial cells, and lymphocytes. Moreover, hetero-cellular spheroids are demonstrated to form more aggregated spheroids and have higher adhesion ability for the mesothelial layer. To improve the poor prognosis, we need to elucidate the mechanisms of spheroid formation and interactions with non-malignant cells in ascites that are a unique tumor microenvironment for EOC.     

Superplastic behaviour of Ti-48at.%Al two-phase titanium aluminide compacts made from mechanically alloyed powder

An HIP compact of MA-processed powder having a nominal composition of Ti-48at.% Al was produced. The compact consisted of a large amount of TiAl(λ) and a small amount of Ti₃Al (₂), in a completely ultra-fine equiaxed grain structure. This two-phase compact showed typical superplastic deformation behaviour. A maximum elongation of 550% was obtained. A strain exponent, n = 2, and grain size exponent, p = 2, were determined from the results of a strain-rate-change test and a creep test at constant initial stress using samples having various grain sizes, respectively. The activation energy for creep, Q_c at constant stress was calculated to be 350 kJ/mole. It is concluded that the superplastic deformation mechanism of the material under study is grain boundary sliding controlled by lattice diffusion in the TiAl phase.     

Genotoxicity of nano/microparticles in in vitro micronuclei, in vivo comet and mutation assay systems

Background

Recently, manufactured nano/microparticles such as fullerenes (C₆₀), carbon black (CB) and ceramic fiber are being widely used because of their desirable properties in industrial, medical and cosmetic fields. However, there are few data on these particles in mammalian mutagenesis and carcinogenesis. To examine genotoxic effects by C₆₀, CB and kaolin, an in vitro micronuclei (MN) test was conducted with human lung cancer cell line, A549 cells. In addition, DNA damage and mutations were analyzed by in vivo assay systems using male C57BL/6J or gpt delta transgenic mice which were intratracheally instilled with single or multiple doses of 0.2 mg per animal of particles.

Results

In in vitro genotoxic analysis, increased MN frequencies were observed in A549 cells treated with C₆₀, CB and kaolin in a dose-dependent manner. These three nano/microparticles also induced DNA damage in the lungs of C57BL/6J mice measured by comet assay. Moreover, single or multiple instillations of C₆₀ and kaolin, increased either or both of gpt and Spi^- mutant frequencies in the lungs of gpt delta transgenic mice. Mutation spectra analysis showed transversions were predominant, and more than 60% of the base substitutions occurred at G:C base pairs in the gpt genes. The G:C to C:G transversion was commonly increased by these particle instillations.

Conclusion

Manufactured nano/microparticles, CB, C₆₀ and kaolin, were shown to be genotoxic in in vitro and in vivo assay systems.     

Random mutagenesis improves the low-temperature activity of the tetrameric 3-isopropylmalate dehydrogenase from the hyperthermophile Sulfolobus tokodaii

In general, the enzymes of thermophilic organisms are more resistant to thermal denaturation than are those of mesophilic or psychrophilic organisms. Further, as is true for their mesophilic and psychrophilic counterparts, the activities of thermophilic enzymes are smaller at temperatures that are less than the optimal temperature. In an effort to characterize the properties that would improve its activity at temperatures less than the optimal, we subjected the thermostable Sulfolobus tokodaii (S. tokodaii) 3-isopropylmalate dehydrogenase to two rounds of random mutagenesis and selected for improved low-temperature activity using an in vivo recombinant Escherichia coli system. Five dehydrogenase mutants were purified and their catalytic properties and thermostabilities characterized. The mutations favorably affect the K(m) values for NAD (nicotinamide adenine dinucleotide) and/or the k(cat) values. The results of thermal stability measurements show that, although the mutations somewhat decrease the stability of the enzyme, the mutants are still very resistant to heat. The locations and properties of the mutations found for the S. tokodaii enzyme are compared with those found for the previously isolated low-temperature adapted mutants of the homologous Thermus thermophilus enzyme. However, there are few, if any, common properties that enhance the low-temperature activities of both enzymes; therefore, there may be many ways to improve the low-temperature catalytic activity of a thermostable enzyme.     

Development of a novel microstructure fabrication method with co-axial dual capillary solution flow type droplet cells and electrochemical deposition

A new method for maskless fabrication of metallic patterns or structures on metals is described. A solution flow type droplet cell, with co-axial dual capillaries was applied to form fine metal structures such as strips and rods. This type of droplet cell enables movement of the cell during formation. Nickel fine patterns with a width of about 200 μm were formed on a Cu substrate. The width of the formed pattern does not change with the scanning speed of the cell, but the thickness of the formed pattern changes with the speed. Two different deposition modes were examined to form metal rods, one is a mold free deposition mode and the other is a mold assisted deposition mode. Both modes enable the formation of Ni rods, however, reproducibility of mold free deposition mode was not good. The mold assisted deposition mode has far better the reproducibility, because of the use of the inside wall of the 100 μm diameter inner tube as the mold. It is possible to form nickel micro-rods, about 100 μm in diameter and 12 mm long with relatively smooth surfaces by the mold assisted deposition mode.     

Thermal and electrical properties of perovskite-type strontium molybdate

Polycrystalline samples of perovskite-type strontium molybdate, SrMoO₃, have been prepared and the thermal and electrical properties have been measured from room temperature to about 1000 K. The electrical resistivity is of an order of magnitude of 10⁻⁵ to 10⁻⁶ (Ω m) in the whole temperature range. The Seebeck coefficient is around 4–9 μV K⁻¹. At room temperature, the thermal conductivity is about 30 W m⁻¹ K⁻¹, and it decreases with increasing temperature.     

The activation of furfuryl alcohol polymerization by oxygen and its enhanced mechanical properties

The effect of oxygen and additional oxygen providers on furfuryl alcohol polymerization was investigated through chemical analyses and mechanical evaluation. NMR, UV–vis, Fourier transform infrared, and gas chromatography–mass spectrometry (GC–MS) results suggested that atmospheric oxygen and the further addition of an oxygen source functioned as an activator for the entire network polymerization. Interestingly, the construction of a conjugated structure on the furan linear chain, which is key to three-dimensional cross-linking, also appears to be accelerated in the presence of oxygen. Furthermore, the introduction of oxygen providers into the curing system successfully enhanced the mechanical properties of the cured furan resin.     

Characterization of a Novel Thermostable Dye-Linked l-Lactate Dehydrogenase Complex and Its Application in Electrochemical Detection

Flavoenzyme dye-linked l-lactate dehydrogenase (Dye-LDH) is primarily involved in energy generation through electron transfer and exhibits potential utility in electrochemical devices. In this study, a gene encoding a Dye-LDH homolog was identified in a hyperthermophilic archaeon, Sulfurisphaera tokodaii. This gene was part of an operon that consisted of four genes that were tandemly arranged in the Sf. tokodaii genome in the following order: stk_16540, stk_16550 (dye-ldh homolog), stk_16560, and stk_16570. This gene cluster was expressed in an archaeal host, Sulfolobus acidocaldarius, and the produced enzyme was purified to homogeneity and characterized. The purified recombinant enzyme exhibited Dye-LDH activity and consisted of two different subunits (products of stk_16540 (α) and stk_16550 (β)), forming a heterohexameric structure (α3β3) with a molecular mass of approximately 253 kDa. Dye-LDH also exhibited excellent stability, retaining full activity upon incubation at 70 °C for 10 min and up to 80% activity after 30 min at 50 °C and pH 6.5–8.0. A quasi-direct electron transfer (DET)-type Dye-LDH was successfully developed by modification of the recombinant enzyme with an artificial redox mediator, phenazine ethosulfate, through amine groups on the enzyme’s surface. This study is the first report describing the development of a quasi-DET-type enzyme by using thermostable Dye-LDH.     

Use of a Metal-mould Process in the Development of Hemispherical Heavy-walled Castings

The exacting quality required of hemispherical heavy-walled steel castings, used in nuclear power applications, has been remarkably improved by the adoption of a newly developed metal-mould process. This technique is dependent on the use of a metal core. The optimum conditions for solidification were determined by the finite element analytical method. The appropriate wall-thickness of the chill elements was also investigated using data based on actual experience. The core was designed in such a manner that the surface carried a series of concave “dimples” in the manner of a golf-ball, the pattern being intended to prevent the formation of cracks, generated on the casting surface when in contact with a metal mould. Stresses caused by solidification shrinkage were reduced by splitting the core. In the event, the casting structure conformed well to the results of the solidification analysis. Radiographic examination revealed a high level of soundness, with no trace of internal defects. No anisotropy was detected in mechanical properties. It is shown that this process is being currently employed in the manufacture of actual products.