Concern over the effects of nanomaterials on human health has risen due to the dramatic advances in the development of various technologies based on nanomaterials. Gifu Prefecture and Gifu University are developing technologies for recycling used carbon fiber because the waste disposal process is highly cost and energy intensive. However, generation of carbon fiber dust during the recycling process is a serious issue, especially in the occupational environment. Recycling requires carbonization by partial firing treatment at 500℃ followed by firing treatment at 440℃: these processes produce dust as a by-product. It is important to study the influence of carbon fibers on human health at a molecular level. In this study, three types of carbon fibers - before recycling, after carbonization, and after firing were evaluated for their toxic effects on mice. During the breeding period, no loss in body weight was confirmed. Further, by staining the lung tissue sections, it was found that pulmonary fibrosis did not occur. We found that these carbon fibers might not possess severe toxicity. However, we also found that the toxicity varies according to firing treatment. Furthermore, we found that firing treatment reduces the potential hazard to human health.

Yttrium oxide (Y₂O₃) nanoparticles have widespread applications; however, toxicity due to these nanoparticles has also been reported. In this study, we evaluated the in vitro toxicity of Y₂O₃ nanoparticles according to the technical specifications published by the International Standard Organization (ISO/TS 19337:2016). We used Saccharomyces cerevisiae as a model microorganism represented the environment. We carried out catch ball analysis of yttrium oxide and yttrium ion toxicities. The result showed that Y₂O₃ nanoparticles (20 mg/5 ml) and YCl₃ (5 mg/5 ml) treatment caused oxidative stress in yeast cells. Based on transcriptome analysis, fluorescent spectroscopy, and solubility analysis of Y₂O₃ nanoparticles, we conclude that the toxicity is due to yttrium ions derived from the nanoparticles. The ions induce oxidative stress and cause protein denaturation, which in turn induces proteasome formation to eliminate denatured proteins. Yttrium nanoparticles induce oxidative stress, which has associated with heavy metal ions. Thus, the use of yttrium nanoparticles or yttrium ions must be controlled like heavy metals.

In this research, we aimed to reveal structures related to reactive oxygen species (ROS) generation of flavonoids upon ultraviolet (UV) or X-ray irradiation. ROS production of selected flavonoids was studied by using ROS fluorescence probes. We discuss the relationship between the structures of the flavonoids and the generation of three ROSs, superoxide anion radical (O-2(center dot-)), hydroxyl radical ((OH)-O-center dot), and singlet oxygen (O-1(2)). Results showed that 2,3-double bond was an key structure for natural flavonoids to generate O-2(center dot-). We also discuss the effects of the ROS scavenging capacities on ROS generation through oxygen radical absorbance capacity (ORAC) tests. Derivatives of quercetin could produce (OH)-O-center dot due to differences of ROS scavenging ability. Our results indicated that flavonoids and their derivatives, which were used as antioxidant reagents, can serve as ROS generators upon ultraviolet (UV) or X-ray irradiation.

Currently, nanoparticles are used in various commercial products. One of the most common nanoparticles is titanium dioxide (TiO2). It has a catalytic activity and UV absorption, and generates reactive oxygen species (ROS). This catalytic activity of TiO2 nanoparticles was believed to be capable of killing a wide range of microorganisms. In the environment, the unique properties of TiO2 nanoparticles can be maintained; therefore, the increasing use of TiO2 nanoparticles is raising concerns about their environmental risks. Thus, assessment of the biological and ecological effects of TiO2-NOAAs is necessary. In this study, we assessed the effect of TiO2-NOAAs for S. cerevisiae using DNA microarray. To compare yeast cells under various conditions, six treatment conditions were prepared (1. adsorbed fraction to TiO2-NOAA under UV; 2. non-adsorbed fraction to TiO2-NOAA under UV; 3. adsorbed fraction to TiO2-NOAA without UV; 4. non-adsorbed fraction to TiO2-NOAA without UV; 5. under UV; and 6. untreated control). The result of the DNA microarray analysis, suggested that yeast cells that are adsorbed by TiO2-NOAA under UV irradiation suffer oxidative stress and this stress response was similar to that by only UV irradiation. We concluded that the effect of TiO2-NOAAs on yeast cells under UV irradiation is not caused by TiO2-NOAA but UV irradiation.

A nanodiamond (ND) is a promising material for drug delivery applications owing to its relatively low cost, amenability to large-scale synthesis, unique structure, and low toxicity. However, synthesizing drug-loaded ND conjugates with uniform and tunable sizes, high loading capacity, efficacy in drug delivery, and versatility in terms of surface functionalization has been challenging. Here, we show that perfluorooctanoic acid-functionalized NDs spontaneously transform into well-dispersed and biocompatible supraparticle (SP) nanoclusters. We demonstrate that the synthesized ND-based SPs (ND-SPs) exhibit high penetration through the cell membrane and are therefore superior as drug carriers for conventional nanomedicines such as polyethylene glycol and phospholipid-based nanocapsules and simple drug-loaded ND conjugates. We confirm the efficacy of ND-SPs in the eradication of cancer cells in vitro and in vivo. Our results demonstrate that the synthesized ND-SPs are useful for targeted drug delivery in a variety of biological applications.

Non-toxic X-ray-responsive substances can be used in the radiosensitization of cancer, like porphyrin mediated radiotherapy. However, most X-ray-responsive substances are toxic. To find novel non-toxic X-ray-responsive substances, we studied the X-ray and UV reactivity of 40 non-toxic compounds extracted from plants. Dihydroethidium was used as an indicator to detect reactive oxygen species (ROS) generated by the compounds under X-ray or UV irradiation. We found that 13 of the investigated compounds generated ROS under X-ray irradiation and 17 generated ROS under UV irradiation. Only 4 substances generated ROS under both X-ray and UV. In particular, luteolin exhibited the highest activity among the investigated compounds; therefore, the ROS generated by luteolin were thoroughly characterized. To identify the ROS, we employed a combination of ROS detection reagents and their quenchers. O-2 generation by luteolin was monitored using dihydroethidium and superoxide dismutase (as an O-2(-) quencher). OH and O-1(2) generation was determined using aminophenyl fluorescein with ethanol (OH quencher) and Singlet Oxygen Sensor Green with NaN3 (O-1(2) quencher), respectively. Generation of O-2(-) under X-ray and UV irradiation was observed; however, no OH or O-1(2) was detected. The production of ROS from luteolin is surprising, because luteolin is a well-known antioxidant.

Combined treatment with 5-aminolevulinic acid (5-ALA) and X-rays improves tumor suppression in vivo. This is because the accumulated protoporphyrin IX from 5-ALA enhances the generation of ROS by the X-ray irradiation. In the present study, a high-energy medical linear accelerator was used instead of a non-medical low energy X-ray irradiator, which had been previously used. Tumor-bearing mice implanted with B16-BL6 melanoma cells were treated with fractionated doses of irradiation (in total, 20 or 30 Gy), using two types of X-ray irradiator after 5-ALA administration. Suppression of tumor growth was enhanced with X-ray irradiation in combination with 5-ALA treatment compared with X-ray treatment alone, using both medical and non-medical X-ray irradiators. 5-ALA has been used clinically for photodynamic therapy. Thus, "radiodynamic therapy", using radiation from medical linacs as a physical driving force, rather than the light used in photodynamic therapy, may have potential clinical applications.

Shochu distillery waste is the discarded material generated during Shochu production, and it has been used as feed for livestock. In this study, we evaluated the applicability of Shochu distillery waste as a livestock diet. Shochu distillery waste diet was fed to Clawn miniature pigs. At the end of the dietary period, blood samples were collected for biochemical examination and microarray analysis. No significant differences were observed between the control and Shochu distillery waste treatment groups based on physical and biochemical examination. Gene expression patterns were also similar. In addition, gene profiling of these two groups was compared with those of hyperlipidemia and toxicant model groups. Expression profiling of the two groups was different from those of the hyperlipidemia and toxicant model groups. In conclusion, the Shochu distillery waste diet did not affect pig physiology and it is a suitable substitute for standard feed. Moreover, these results promote the potential for microarray analysis use in the evaluation of food safety.

Purpose: 5-Aminolevulinic acid (ALA) is a precursor of the photosensitizer protoporphyrin (PpIX) used in photodynamic therapy. In our previous work, PpIX enhanced the generation of reactive oxygen species by X-ray irradiation. In this study, we evaluated the potential of ALA as an endogenous sensitizer to X-ray irradiation.
Methodology: Tumor-bearing C57BL/6J mice implanted with B16-BL6 melanoma cells were subsequently treated with irradiation (3Gy/day for 10 days; total, 30Gy) plus local administration of 50mg/kg ALA 24 hours prior to each irradiation (ALA-XT). Tumor-bearing mice without treatment (NT), those treated with ALA only (ALAT), and those treated with X-ray irradiation only (XT) were used as controls.
Results: ALA potentiated tumor suppression by X-ray irradiation. In microarray analyses using tumor tissue collected after 10 sessions of fractional irradiation, functional analysis revealed that the majority of dysregulated genes in the XT and ALA-XT groups were related to cell-cycle arrest. Finally, the XT and ALA-XT groups differed in the strength of expression, but not in the pattern of expression.
Conclusions: mRNA analysis revealed that the combined use of ALA and X-ray irradiation sensitized tumors to X-ray treatment. Furthermore, the present results were consistent with ALA's tumor suppressive effects in vivo.

Protoporphyrin IX (PpIX) enhances the generation of reactive oxygen species in cells following physicochemical interactions with X-rays. To evaluate the use of porphyrins as radio-sensitizers in radiotherapy, the transcriptomic effects of PpIX and/or X-ray irradiation were investigated in HeLa cells. Microarray experiments were performed using Agilent-014,850 Whole Human Genome Microarray 4x44K G4112F (GEO#: GSE61805). We selected the condition corresponding to 1 μg/mL PpIX exposure prior to 3 Gy-irradiation of cells, and collected cells 24 h post irradiation. X-ray exposure at a dose of 3 Gy did not affect cell survival 24 h post irradiation, regardless of the concentration of PpIX. Approximately 50% cells exposed to X-ray irradiation alone (XT) and 70% cells exposed to PpIX treatment for 6 h before X-ray irradiation (PpIX-XT) lost clonogenic ability. Based on p-values (p < 0.01), we selected genes for functional analysis. The majority of the regulated genes in the XT and PpIX-XT groups were related to cell cycle arrest. Furthermore, transcriptome analysis of the cells collected 24 h post irradiation revealed the fate of the cells that lost clonogenic ability due to cell cycle arrest.

5-Aminolevulinic acid (ALA) is a photosensitizer used in photodynamic therapy (PDT) because it causes preferential accumulation of protoporphyrin IX (PpIX) in tumor cells, where it forms singlet oxygen upon light irradiation and kills the tumor cells. Our previous study demonstrated that PpIX enhances generation of reactive oxygen species by physicochemical interaction with X-rays. We investigated the effect of ALA administration with X-ray irradiation of mouse B16-BL6 melanoma cells in vitro and in vivo. ALA facilitates PpIX accumulation in tumor cells and enhances ROS generation in vitro. Tumor suppression significantly improved in animals treated with fractionated doses of radiation (3 Gy x 10; total, 30 Gy) with local administration of 50 mg/kg ALA at 24 h prior to fractional irradiation. These results suggest ALA may improve the efficacy of cancer radiotherapy by acting as a radiomediator.

Background: Hyperlipidemia animal models have been established, but complete gene expression profiles of the transition from normal lipid levels have not been obtained. Miniature pigs are useful model animals for gene expression studies on dietary-induced hyperlipidemia because they have a similar anatomy and digestive physiology to humans, and blood samples can be obtained from them repeatedly.
Methodology: Two typical dietary treatments were used for dietary-induced hyperlipidemia models, by using specific pathogen-free (SPF) Clawn miniature pigs. One was a high-fat and high-cholesterol diet (HFCD) and the other was a high-fat, high-cholesterol, and high-sucrose diet (HFCSD). Microarray analyses were conducted from whole blood samples during the dietary period and from white blood cells at the end of the dietary period to evaluate the transition of expression profiles of the two dietary models.
Principal Findings: Variations in whole blood gene expression intensity within the HFCD or the HFCSD group were in the same range as the controls provide with normal diet at all periods. This indicates uniformity of dietary-induced hyperlipidemia for our dietary protocols. Gene ontology-(GO) based functional analyses revealed that characteristics of the common changes between HFCD and HFCSD were involved in inflammatory responses and reproduction. The correlation coefficient between whole blood and white blood cell expression profiles at 27 weeks with the HFCSD diet was significantly lower than that of the control and HFCD diet groups. This may be due to the effects of RNA originating from the tissues and/or organs.
Conclusions: No statistically significant differences in fasting plasma lipids and glucose levels between the HFCD and HFCSD groups were observed. However, blood RNA analyses revealed different characteristics corresponding to the dietary protocols. In this study, whole blood RNA analyses proved to be a useful tool to evaluate transitions in dietary-induced hyperlipidemia gene expression profiles in miniature pigs.

The radiosensitizing effect of 5-250 nm diameter Au nanoparticles (AuNPs) in water was investigated under irradiations of diagnostic x-ray and UV light. Enhanced generations of hydroxyl radical ((center dot)OH) and superoxide anion (O(2)(-)) were confirmed from their dependencies on the absorbed energy, ethanol concentration and AuNPs&apos; concentration. Two kinds of fluorescent probes revealed that the reactive oxygen species (ROS) generation rate under x-ray irradiation was enhanced by factors of 1.46 for (center dot)OH and 7.68 for O(2)(-). Photo-and Auger electron charge transfer is possibly relevant to generation of O(2)(-) near the particle surface, whereas fluorescent x-rays are involved in generation of (center dot)OH in the secondary water radiolysis. Smaller diameter AuNPs with larger surface area showed a greater yield of ROS. An inverse proportion of ROS generation to the AuNPs&apos; diameter suggests a catalytic function of AuNPs&apos; surface for enhanced ROS generation.
From the Clinical Editor: This paper investigates the effects of UV and X-ray irradiation on reactive oxygen species induction of gold nanoparticles, concluding that smaller diameter AuNPs with larger surface area lead to a greater yield of ROS probably due to catalytic effects. The paper may be important for the development of novel non-toxic radiation sensitizers. (C) 2011 Elsevier Inc. All rights reserved.

Miniature pigs are useful model animals for humans because they have similar anatomy and digestive physiology to humans and are easy to breed and handle. In this study, whole blood microarray analyses were conducted to evaluate variations of correlation among individuals and ages using specific pathogen-free (SPF) Clawn miniature pigs. Whole blood RNA is easy to handle compared to isolated white blood cell RNA and can be used for health and disease monitoring and animal control. In addition, whole blood is a heterogeneous mixture of subpopulation cells. Once a great change occurs in composition and expressing condition of subpopulations, their associated change will be reflected on whole blood RNA. From 12 to 30 weeks of age, fractions of lymphocytes, monocytes, neutrophils, eosinophils, and basophils in white blood cells showed insignificant differences with age as a result of ANOVA analysis. This study attempted to identify characteristics of age-related gene expression by taking into account the change in the number of expressed genes by age and similarities of gene expression intensity between individuals. As a result, the number of expressed genes was less in fetal stage and infancy period but increased with age, reaching a steady state of gene expression after 20 weeks of age. Variation in gene expression intensity within the same age was great in fetal stage and infancy period, but converged with age. The variation between 20 and 30 weeks of age was comparable to that among 30 weeks individuals. These results indicate that uniformity of laboratory animals is expected for miniature pigs after 20 weeks of age. Furthermore, a possibility was shown that whole blood RNA analysis is applicable to evaluation of physiological state.

As a possible radiosensitizer candidate having biological compatibility, oncotropic property, and X-ray activation capability, contribution of protoporphyrin IX (PpIX) to enhanced generation of reactive oxygen species (ROS) under X-ray and UV irradiations were examined. To identify the kinds of ROS, 2-[6-(4-amino)phenoxy-3H-xanthen-3-on-9-yl] benzoic acid (APF) and dihydroethidium (DHE) were used together with ethanol as a hydroxyl radical (OH(center dot)) quencher. All of the three species of our interest (OH(center dot), superoxide radical (O(2)(center dot-)), and singlet oxygen ((1)O(2))) were enhanced by PpIX under X-ray and UV irradiations in addition to those by radiolysis and photolysis. Its enhancement factors exceeded 1.7 depending on the concentrations of PpIX from 1.5 to 15.0 mu g/ml. (C) 2009 Elsevier Ltd. All rights reserved.

The research field of high pressure processed food technology was spread from basic science to commercial basis progress. Thus, this technology is being accepted by consumer and is now on the stage of standardization. Standardization is required for the stable and safety supply of high pressure processed food to consumer. In this paper, we would like to suggest the draft version of &quot;Technical Report&quot; concern to indicative standard microorganisms using yeast, &lt;I&gt;Saccharomyces cerevisiae&lt;/I&gt;. We selected the strain of S288C, medium of YPD, and cultivation time of 50-150h at 25°C. The strain was selected as the genome sequence of this strain was already analyzed and thus the most characterized strain in the yeast strains. YPD medium is widely used by basic scientist and applied scientist. The cultivation time was examined by counting CFU after and before pressure treatment and we found yeast cells during this growth phase showed the reproducible high pressure resistance. Next step is the validation studies of this &quot;Technical Report&quot; in different laboratories. We would like to call for participation in the validation studies to laboratories.

For a development of deep tumor treatment in photodynamic therapy, a feasibility of novel radiosensitizers induced by x-ray was investigated. The sensitizers are designed to generate reactive oxygen species (ROS) inside or outside the cell, possibly leading to damage exclusively on tumor cells and reservation of normal cells along the x-ray path. Taking note of the similarity in energy transfer mechanism in photocatalysts, scintillators, and particulate semiconductors, we chose TiO2, ZnS:Ag, CeF3, and quantum dots (CdTe and CdSe) in particulate form, which contain heavy atoms for efficient absorption of x-rays. A parameter study for x-ray operating conditions showed that in a typical scenario, photons with 20 to 170 keV energy are attenuated by 90% through the region of particle dispersed aqueous solution at varying concentration between 0.01 and 100 wt%. The amount of ROS generation under the exposure of polychromatic x-ray was measured using dihydroethidium reagent which detects an integrated amount of several species. Proportional increase in ROS generation to x-ray dose was observed for varying concentrations of TiO2, ZnS:Ag, CeF3, and CdSe quantum dot dispersions. Then, HeLa cells were mixed with aqueous solutions dispersed with sensitizing materials at a concentration of 3.0 mg/ml and were exposed to x-ray. Their survival fraction obtained by a cell proliferation reagent WST-1 immediately after the irradiation showed insignificant effects of sensitizing materials except at large doses. To enhance the sensitization effect, bio-conjugated CdSe quantum dots were internalized in the cytoplasm up to a concentration of 1.0 ng/ml. The cells were irradiated by x-ray up to 5 Gy, and their survival fraction was measured by the colony forming ability 9 days after irradiation. Survival fraction of the cells treated with quantum dots were less than those without quantum dots for all doses, suggesting that the colony forming ability is impaired by the internalized quantum dots. © 2008 Humana Press Inc.

To realize highly integrated micro total analysis systems (mu TAS), a simply controlled miniaturized valve should be utilized on microfluidic device. In this paper, we describe the application of photo-induced super-hydrophilicity of titanium dioxide (TiO2) to microfluidic manipulation. In addition, we found a new phenomenon for reversibly converting the surface wettability using a polydimethylsiloxane (PDMS) matrix and the photocatalytic properties of TiO2. While PDMS polymer was irradiated with UV, it was confirmed that hydrophobic material was released from the polymer to air. Several prepolymers were identified as the hydrophobic material with a gas chromatograph and mass spectrometer (GUMS). Here, we successfully demonstrated the flexible manipulation of microfluid in a branched microchannel using the reversible wettability as micro opto-switching valve (MOS/V). The simultaneous control of MOS/Vs was also demonstrated on a 256-MOS/V integrated disk. The MOS/V promises to be one of the most effective flow switching valves for advanced applications in highly integrated micro/nano fluidics. (c) 2006 Elsevier B.V. All rights reserved.

Plant-derived essential oils with monoterpenoids have been used as antifungal drugs since ancient times, but the mode of action of these natural hydrocarbons at the molecular level is not understood. In order to understand the mechanisms of toxicity of alpha-terpinene (a cyclic monoterpene), a culture of Saccharomyces cerevisiae was exposed to 0.02% alpha-terpinene for 2 h and transcript profiles were obtained using yeast DNA arrays. These profiles, when compared with transcript profiles of untreated cultures, revealed that the expression of 793 genes was affected. For 435 genes, mRNA levels in treated cells compared with control cells differed by more than two-fold, whereas for 358 genes, it was &lt;0.5-fold. Northern blots were performed for selected genes to verify the microarray results. Functional analysis of the up-regulated genes indicates that, similar to commonly used antifungal drugs, alpha-terpinene exposure affected genes involved in ergosterol biosynthesis and sterol uptake. In addition, transcriptional induction of genes related to lipid metabolism, cell wall structure and function, detoxification and cellular transport was observed in response to terpinene toxicity. Notably, the functions of 192 up-regulated genes are still unknown, but their characterization will probably shed light on the mechanisms of drug resistance and sensitivity. Taken together, this study showed that alpha-terpinene has strong antifungal activities and its modes of action resemble those of presently used antifungal drugs.

Although there have been studies on the toxicity of the pesticide thiuram, the present study is the first one to attempt to integrate a whole genomic response using microarray technology. From the DNA microarray experiment, it was found that exposure to thiuram led to alterations of gene expression in yeast cells and that many genes involved in detoxification and stress response were highly induced. The induced genes were classified according to the MIPS yeast database. The induction of genes concerned with folding and proteolysis reflects the protein denaturing and degradation effects of the thiuram treatment. The induction of genes involved in redox and defense against reaction oxygen species also suggests that thiuram has other effects, such as oxidative stress. Genes classified for carbohydrate metabolism and energy were also highly induced, and these gene products may play the role of providing the energy for the detoxification mechanism. In addition, in view of the induction of some genes involved in DNA repair, thiuram potentially causes DNA damage. Therefore, as stated in previous reports, thiuram is a potential positive toxic chemical. On the other hand, YKL071W, YCR102C, YLR303W, and YLL057C were selected based on the result of a DNA microarray experiment and used for the promoter activity assay. Thiuram treatment affected the promoter of these genes, indicating that this technique could be used for the selection of biomarker candidates.

Characteristics of solute removal and biocompatibility were clinically compared among three kinds of membrane hemodialyzers with minimum activating capacity for complement, and comparable in vitro clearance of urea. Ten patients with chronic renal failure who had undergone hemodialysis (HD) three times a week for more than one year were enrolled in this study. These patients were dialyzed using three kinds of dialyzer membranes, Hemophan (H), cellulose di-acetate (CA), and polymethylmethacrylate (PMMA) in a crossover method under HD conditions that were similer except for dialyzer membrane. Testing of HD included evaluation of clearance and reduction rate of small molecular substances (urea nitrogen, UN; creatinine, Cr ; uric acid, UA ; and phosphate, iP) ; and changes of blood cell counts ; beta-thromboglobulin (β-TG) ; thromboxane B₂; 6-keto-prostaglandin F1α; granulocyte elastase-α_l proteinase inhibitor complex (G-elastase) ; celite activated coagulation time (CCT) ; and fibrinopeptide A (FPA) . Although the clearance and reduction rates of UN, Cr and UA did not differ among the three membranes, those of iP were significantly lower with PMMA than with CA or H, and the clearance of iP was maximum with H. HD with H induced a greater decrease of leucocytes, than with CA and a higher venous/arterial ratio of β-TG than CA or PMMA. HD with H prolonged CCT less than HD with PMMA, and increased FPA more than CA or PMMA. The largest increase in G-elastase was observed in HD with PMMA. Strongly negative, and relatively positive membrane charges were reported in PMMA and H, respectively. It is well known that positive membrane charge is an activating stimulus for platelets. The results thus suggest that elimination of negatively charged iP decreased in HD with PMMA and increased with H membrane because of membrane charges, and that coagulation cascade was stimulated during HD with H, possibly because its positive membrane charge activated platelets and adsorbed negatively charged heparin.