Production of cordycepin by a repeated batch culture of a Cordyceps militaris mutant obtained by proton beam irradiation

Cordycepin (3′-deoxyadenosine) is one of the most versatile metabolites of Cordyceps militaris due to its broad spectrum of biological activity. In our previous study, the C. militaris mutant G81-3, which produces higher levels of cordycepin, was obtained by high-energy proton beam irradiation. In this study, the effects of adenosine on cordycepin production in a surface liquid culture of the mutant and the wild type strains were investigated. For the mutant strain, the optimum dose of adenosine yielded a 30% increase in cordycepin production; the maximum levels of production with adenosine and without adenosine were 8.6 g/l and 6.7 g/l, respectively. In contrast, the increase due to adenosine supplementation for the wild type strain was only 15% (3.1 g/l with adenosine and 2.7 g/l without adenosine). Furthermore, a repeated batch culture, an efficient production method, was carried out to eliminate the relatively long lag phase of the mutant culture. Over four cycles, both the mutant and the wild type strain maintained a production level of more than 85% of that of the initial cycle. As a result, the disadvantage of the mutant was successfully overcome, resulting in a productivity (0.48 g/(l d)) higher than that of the batch culture (0.29 g/(l d)). The productivity for cordycepin obtained in this study is the highest reported value to date, and this method could be applied to large-scale production of cordycepin at industrial levels.     

Self-assemblies of linearly aligned diamond fillers in polysiloxane/diamond composite films with enhanced thermal conductivity

The relocation of diamond fillers was performed in polysiloxane-based composite films under different electric fields. The microscale diamond filler particles were dispersed by sonication in a prepolymer mixture of polysiloxane, followed by high-speed mixing. The homogeneous suspension was cast onto a polyamide spacer of microscale thickness and subjected to three different electric fields: AC, DC, and switched DC, before the mixture became cross-linked. Analysis revealed that self-assemblies of linearly aligned diamond fillers (LADFs) were fabricated in the composite film, connecting the film planes as bridges with different thicknesses depending on the applied electric field. Composites with assemblies of LADFs exhibited enhanced thermal conductivity and electrical insulation, and are attractive for application as thermal interface materials in the semiconductor industry.     

Synergetic Effects of PARP Inhibitor AZD2281 and Cisplatin in Oral Squamous Cell Carcinoma in Vitro and in Vivo

Cisplatin is a commonly used chemotherapeutic drug for treatment of oral carcinoma, and combinatorial effects are expected to exert greater therapeutic efficacy compared with monotherapy. Poly(ADP-ribosyl)ation is reported to be involved in a variety of cellular processes, such as DNA repair, cell death, telomere regulation, and genomic stability. Based on these properties, poly(ADP-ribose) polymerase (PARP) inhibitors are used for treatment of cancers, such as BRCA1/2 mutated breast and ovarian cancers, or certain solid cancers in combination with anti-cancer drugs. However, the effects on oral cancer have not been fully evaluated. In this study, we examined the effects of PARP inhibitor on the survival of human oral cancer cells in vitro and xenografted tumors derived from human oral cancer cells in vivo. In vitro effects were assessed by microculture tetrazolium and survival assays. The PARP inhibitor AZD2281 (olaparib) showed synergetic effects with cisplatin in a dose-dependent manner. Combinatorial treatment with cisplatin and AZD2281 significantly inhibited xenografted tumor growth compared with single treatment of cisplatin or AZD2281. Histopathological analysis revealed that cisplatin and AZD2281 increased TUNEL-positive cells and decreased Ki67- and CD31-positive cells. These results suggest that PARP inhibitors have the potential to improve therapeutic strategies for oral cancer.     

Individual Variations in Inorganic Arsenic Metabolism Associated with AS3MT Genetic Polymorphisms

Individual variations in inorganic arsenic metabolism may influence the toxic effects. Arsenic (+3 oxidation state) methyltransferase (AS3MT) that can catalyze the transfer of a methyl group from S-adenosyl-l-methionine (AdoMet) to trivalent arsenical, may play a role in arsenic metabolism in humans. Since the genetic polymorphisms of AS3MT gene may be associated with the susceptibility to inorganic arsenic toxicity, relationships of several single nucleotide polymorphisms (SNPs) in AS3MT with inorganic arsenic metabolism have been investigated. Here, we summarize our recent findings and other previous studies on the inorganic arsenic metabolism and AS3MT genetic polymorphisms in humans. Results of genotype dependent differences in arsenic metabolism for most of SNPs in AS3MT were Inconsistent throughout the studies. Nevertheless, two SNPs, AS3MT 12390 (rs3740393) and 14458 (rs11191439) were consistently related to arsenic methylation regardless of the populations examined for the analysis. Thus, these SNPs may be useful indicators to predict the arsenic metabolism via methylation pathways.     

Low-Temperature Synthesis of Nanocrystalline Zinc Titanate Materials with High Specific Surface Area

Nanocrystalline zinc titanate (ZnTiO₃) was synthesized at low temperatures through the combination of a sol–gel processing and a polymer binder method. ZnTiO₃ powders of ∼5 nm in size were obtained by heating pastes, which were composed of a Zn-Ti methanolic solution containing acetylacetone and an organic polymer binder, at 500°C in air. Thermal decomposition behavior of the pastes was analyzed by thermogravimetry-differential thermal analysis. Crystallinity of ZnTiO₃ was examined by transmission electron microscopy. The BET measurement revealed that the powders had a relatively high specific surface area of 106 m²/g.     

InAlAs/InGaAs channel composition modulated transistors with InAschannel and AlAs/InAs superlattice barrier layer

We have successfully fabricated a new type of InAlAs/InGaAs heterojunction FET (HJFET) with modulated indium composition channels, called channel composition modulated transistors (CCMTs) in which an InAs channel is sandwiched by In_0.53Ga_0.47As/In _0.8Ga_0.2As sub-channels. The fabricated devices also employ an AlAs/InAs superlattice as a barrier layer against impurity contamination to provide high thermal stability. A 0.2-μm T-shaped gate device exhibits a g_m of 1370 mS/mm, F_t of 180 GHz and F_max of 210 GHz at a low drain bias of 1.0 V. In high-temperature DC life tests conducted at more than 230°C, the devices exhibited less than a 3% degradation in I_dss and g_m after 1000 h. This demonstrates that these newly-developed CCMTs incorporating AlAs/InAs superlattice insertion technology can offer high-performance and highly-reliable InP-based HJFET's for various microwave and millimeter-wave applications     

Hepatocyte growth factor (HGF) produced by peritoneal fibroblasts may affect mesothelial cell morphology and promote peritoneal dissemination

Mesothelial cell monolayers have been reported to prevent infiltration of cancer cells into the peritoneum. We have previously reported that peritoneal fibrosis induced by gastric cancer cells prior to metastatization may provide a congenial environment for peritoneal metastases. In this study, we investigated the effects of peritoneal fibroblasts on peritoneal mesothelial cell morphology. Human gastric cancer (OCUM-2MD3), peritoneal fibroblast (NF-2P) and mesothelial (MS-1) cell lines were established in our laboratory. Histology of the peritoneum was investigated following intraperitoneal inoculation of serum-free conditioned media (SF-CM) from OCUM-2MD3 cells into nude mice. SF-CM from peritoneal fibroblasts was added to monolayer-cultured mesothelial cells, and their morphology was examined by phase-contrast microscopy. This experiment was conducted in the presence and absence of neutralizing antibodies against various factors. Mesothelial cells exposed to fibroblasts proliferation became hemispherical and separated from each other, while unexposed mesothelium remained as a flat monolayer. Cultured-mesothelial cells rounded up or exhibited a fibroblast-like shape following the addition of peritoneal fibroblast SF-CM. Anti-hepatocyte growth factor (HGF) neutralizing antibody partly inhibited this effect. We suggest that soluble factors, such as HGF, produced by peritoneal fibroblasts affect the morphology of mesothelial cells in monolayers so that the resulting environment may become prone to the peritoneal dissemination of cancer cells.     

Effects of liposome-encapsulated hemoglobin on phorbol ester-induced superoxide production and expression of costimulatory molecules by monocytes in vitro

The effects of Neo Red Cell (NRC), a liposome-encapsulated hemoglobin (LEH), on the phorbol ester-induced superoxide production and the expression of costimulatory molecules by human peripheral monocytes were investigated. The treatment of human mononuclear cells with NRC caused the potentiation of superoxide production in response to PMA. The longer incubation (20 h) resulted in a decrease in the PMA-induced superoxide production, which was in parallel to a decrease in the viability of the monocytes. A flow cytometric analysis showed that a slight expression of CD80 (B7-1) on monocytes was induced by NRC treatment, whereas the constitutive expressions of CD86 (B7-2) and CD54 (ICAM-1) were unchanged. The activation of monocytes with interferon-gamma (IFN-gamma) induced the expressions of CD80, CD86, and CD54 under all conditions tested, but NRC treatment tended to decrease the IFN-gamma-induced expression of CD54 on monocytes. These results suggest that the administration of LEH may modify the functions of human monocytes.