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Structural colors with tunable properties have extensive applications in surface decoration, arts, absorbers, and optical filters. Planar structures have more advantages over other forms studied to date due to their easy manufacturability. Metal-insulator-metal-based structures are one of the known methods to fabricate structural colors where colors can be tuned mainly by the thickness of the intermediate lossless insulator layer. However, generating colors by MIM structure requires a thin metallic layer on top, and the top metals’ abrasiveness and/or oxidation may degrade the colors quickly. Thus, we propose a lossy dielectric layer to replace the top metallic layer as a solution to ensure the structure’s durability by preventing scratches and oxidation. Herein, CrON/Si3N4/Metal structures have been studied where theoretical investigations suggest that highly saturated colors can be generated in the lossy-lossless dielectric structures. Experimental data validated such simulations by revealing a range of vivid colors. Furthermore, these structures can easily achieve strong light absorption (SLA) even for a thick top layer of ∼100 nm. The colors realized by these structures are appeared due to a combination of the interference effect of the asymmetric Fabry–Perot cavity structure and the absorption rate in the CrO x N1−x layer.

In this study, an analytical method was established and validated to determine the preservatives such as dehydroacetic acid, benzoic acid, sorbic acid, methylparaben and ethylparaben. The level of preservatives was measured by solvent extraction method adding purification process with carrez reagent and by high-performance liquid chromatography (HPLC). The developed analytical method was successfully applied to determine the concentration of preservatives in various food samples including jam, cheese and soy sauce, displaying high accuracy (recoveries between 87.8% and 110%) and precision (%RSD less than 5.92% and 7.72% for intra-day and inter-day, respectively). To verify the applicability of the improved test method, selected 13 food items and collected 521 samples were monitored. As a result, all the cases met the Korea standard guidelines. Consequently, this study is expected to contribute to the safety management of preservatives for domestic distribution and imported food.

In this study, moth-eye-inspired structures (MISs), which improve optical performance of surfaces, were developed to enhance light efficiency in various optical fields. As the polymer-based soft lithography process can easily optimize MIS materials and nanostructures, 12 MIS/inverse MIS (IMIS) surfaces of 300, 500, and 1000 nm composed of polyurethaneacrylate (PUA) and polydimethylsiloxane (PDMS) polymers were fabricated, and a thin octafluorocyclobutane (C4F8) layer was deposited on the surfaces to reduce their surface energies. The results showed that the 300-nm PDMS-MIS showed excellent optical properties with 94.44 % transmittance and 3.28 % reflectance. Additionally, two theoretical phenomena, the diffraction effect and double-slit diffraction, were experimentally demonstrated using three single-wavelength lasers. Subsequently, contact-angle measurements demonstrated all MIS/IMIS surfaces to possess hydrophobic and superhydrophobic properties, which can protect devices from outdoor liquids. Finally, we confirmed the applicability of MIS by attaching it to a cell phone display and an ITO glass, which is used as a substrate in various optical instruments.

We report a theoretical investigation of field emission from metals in n-dimension. The field-emission current density is derived as a function of the electric field in one to three dimensions. Then, the generalized field-emission current density is calculated for arbitrary n dimensions, is shown as a function of electric field for different work functions of one-, two- and three-dimensional materials, and is expressed as a function of dimension for different work functions. These analyses reveal that the current density increases as the spatial dimension increases in the conductors and work function decreases. Our study on the generalized current density can be applied to material systems with high dimensions or fractional dimensions.

To realize the chip-scale atomic clocks (CSACs) for atomic reference with miniaturization and low power consumption, there are technological challenges in the physics package for compact and stable frequency standards. Here, the precise thermal management and the exact magnetic-field control of the physics package are required for accurate clocks. Also, high thermal resistance for the physics package helps to considerably reduce power consumption of the CSAC. Thus, in this study, a compact physics package with embedded magnetic shielding has been developed by using flexible hybrid approach for a chip-scale atomic clock. The vertical-cavity surface-emitting laser (VCSEL) and the MEMS hotplate on a flexible circuit board (FPCB) are vertically stacked and bound with a QWP and a MEMS vapor cell by aluminum foil. The aluminum bound structure is suspended by the polyimide core of the FPCB for high thermal resistance. Also, the mu-metal fixture is applied for magnetic shield and FPCB suspension, while the MEMS hotplate with a resistance heater and an amorphous Si based thermistor is adopted for thermal management of the physics package. Then, the planar integrated FPCB is folded and stacked vertically at an intersection with the mu-metal fixture. As a result, the CSAC incorporating the proposed physics package reveals the CPT linewidth of 5.6 kHz and low power consumption of about 200mW. The measured frequency short-term stability is about $5.2\times 10^{-12} \tau ^{-1/2}$ at 1000-second integration. The observed results demonstrate promising performances of the flexible hybrid physics package for its use in miniaturized atomic clocks.

Randomly distributed plasmonic Ag nanoparticles (NPs) with various sizes were fabricated by a reflow process to an island-shaped Ag thin-film deposited on a Si photodiode. These NPs conformally enclosed by an antireflective (AR)-type SiNx/SiO2 bilayer reveal significantly diminished reflectance in a broad wavelength (500 nm - 1100 nm) as compared to the cases of Ag NPs or SiO2 layer enclosing Ag NPs on the Si substrate. Accordingly, the forward scattering and the total reflection along with wide-angle interference in between the dielectric bilayer incorporating the Ag NPs induce highly increased light absorption in the Si substrate. The fabricated Si photodiode adopting the plasmonic AR bilayer shows the responsivity peak value of 0.72 A/W at 835 nm wavelength and significant responsivity enhancement up to 40% relative to a bare Si photodiode in a wavelength range of 500 nm to 1000 nm.

Familial adenomatous polyposis (FAP) is an autosomal dominant hereditary cancer syndrome characterized by hundreds to thousands of colorectal adenomatous polyps. Without treatment, progression to colorectal cancer is inevitable. Most pathogenic mutations in the APC gene were nonsense or frameshift mutations; however, some previous studies reported large deletions or duplications.We reviewed the results of APC gene analyses from January 2010 to December 2020. We analyzed the entire coding sequence of the APC gene by Sanger sequencing to detect genetic abnormalities. Moreover, multiplex ligation-dependent probe amplification (MLPA) testing was performed starting in September 2017, and a multigene panel study that includes the APC gene was begun in July 2019.In the 266 collected cases, pathogenic variants/likely pathogenic variants (PV/LPVs) in the APC gene were detected in 73 patients, and variants of uncertain significance were found in 13 patients. Among those variants, 14 variants were novel. We performed MLPA for 29 patients, and 7 of them (24.1%) were positive for a large duplication/deletion. Among the 73 cases in the multigene panel study, 17 cases (23.3%) of APC gene variation were detected.We retrospectively analyzed the APC gene in Korean patients suspected to have FAP. Variants truncated by nonsense and frame shift mutations were common PV/LPVs. However, the detection rate in the MLPA study was higher than in previous studies of Caucasian populations. We suggest that MLPA should be performed for patients likely to have FAP but in whom no variant is detected by sequencing methods.

To realize high-resolution thermal images with high quality, it is essential to improve the noise characteristics of the widely adopted uncooled microbolometers. In this work, we applied the post-metal annealing (PMA) process under the condition of deuterium forming gas, at 10 atm and 300 °C for 30 min, to reduce the noise level of amorphous-Si microbolometers. Here, the DC and temperature coefficient of resistance (TCR) measurements of the devices as well as 1/f noise analysis were performed before and after the PMA treatment, while changing the width of the resistance layer of the microbolometers with 35 μm or 12 μm pixel. As a result, the microbolometers treated by the PMA process show the decrease in resistance by about 60% and the increase in TCR value up to 48.2% at 10 Hz, as compared to the reference device. Moreover, it is observed that the noise characteristics are improved in inverse proportion to the width of the resistance layer. This improvement is attributed to the cured poly-silicon grain boundary through the hydrogen passivation by heat and deuterium atoms applied during the PMA, which leads to the uniform current path inside the pixel.

A simple ionic chromatography method for nitrite analysis in processed food products was developed and validated. Nitrite in the sample was extracted using 80 °C distilled deionized water and centrifuged. Purification of nitrite from sample solution was performed using OnGuard II Ag, OnGuard II RP and OnGuard II Na cartridge connected in order. Determination of nitrite was carried out using IonPac AG9-HC (4 × 50 mm) and IonPac AS9-HC (4 × 250 mm) columns and a 9 mM sodium carbonate mobile phase. The validated results showed good linearity (r

The effect of nanobump structure (NS) constructed with molybdenum oxide (MoO3) covering aerosol-derived Ag nanoparticles on organic solar cells is systematically explored by varying the MoO3 thickness. The amount of enhanced light absorption by the NS-induced scattering and plasmonic resonance effects increases with decreasing MoO3 thickness. Meanwhile, the oval-shape NS with fully enclosed Ag nanoparticles by MoO3 leads to significantly improved carrier extraction due to decreased ohmic loss, while the NS with partially covered Ag nanoparticles by thin MoO3 degrades device performance arising from increased recombination and leakage loss. Thus, the optimal MoO3 thickness occurs around 20 nm for 40 nm sized nanoparticles where the enhanced carrier generation and the improved carrier extraction by the NS effectively contribute to the improvement in photocurrent. Therefore, our analysis on carrier dynamics of plasmonic organic solar cells incorporating the NS would provide a clear guideline for optimizing device performance.

Thermionic emission for quasi-free electron metals is investigated in arbitrary dimension. The thermionic emission for a constant temperature and work function is calculated for one, two and three dimensions. The generalized current density of the thermionic emission is calculated for arbitrary dimension. The current density is shown as a function of temperature for different work functions. The current density is also shown as a function of dimension for different temperatures and work functions.

Silicon dioxide nanoparticles (SiO2-NPs) are widely used in biomedicines and consumer products, such as sunscreens and cosmetics. However, SiO2-NPs can cause adverse effects on human health, depending on the size and concentration of nanoparticles. The present study was aimed at investigating the molecular mechanism underlying SiO2-NPs-induced inflammation in human keratinocyte (HaCaT) cells. Incubation of HaCaT cells with SiO2-NPs induced the expression of cyclooxygenase-2 (COX-2) mRNA and protein. Treatment of cells with SiO2-NPs also induced the phosphorylation, DNA binding and the reporter gene activity of signal transducer and activator of transcription 3 (STAT3). Transfection of cells with STAT3 siRNA abrogated SiO2-NPs-induced COX-2 expression. Moreover, SiO2-NPs enhanced the phosphorylation of Janus kinase2 (JAK2), Src and Akt. Pharmacological inhibition of either JAK2, Src or Akt abrogated SiO2-NPs-induced STAT3 transcriptional activity and the expression of COX-2. Treatment with LY294002 also attenuated SiO2-NPs-induced Src phosphorylation, while, JAK2 phosphorylation was not changed. In addition, SiO2-NPs generated reactive oxygen species (ROS) and treatment of N-acetyl cysteine (NAC) attenuated the phosphorylation of JAK2, Src, Akt and STAT3, as well as the expression of COX-2 in SiO2-NPs-treated HaCaT cells. Taken together, our study provides the first report that SiO2-NPs induce COX-2 expression in HaCaT cells by activating the STAT3 signaling through ROS-mediated phosphorylation of upstream kinases, Akt/Src and JAK2.

In vivo phototoxicity testing is important for predicting drug-induced phototoxicity in humans. Currently, there is no internationally validated in vivo test method for the photosafety evaluation of pharmaceuticals. In this study, we evaluated the phototoxicity of systemically administered drugs using SD rats. We first determined the appropriate ultraviolet A (UVA) dose using 8-methoxypsoralen, a well-known phototoxic drug. Compared to lower and higher UVA doses, we found that a UVA dose of 10 J/cm(2) allowed for the detection of phototoxic responses in both a dose- and time-dependent manner. We next performed a phototoxicity study using seven pharmaceutical drugs which included known phototoxic and non-phototoxic drugs using a UVA dose of 10 J/cm(2). In order to improve the accuracy of our assessment, we evaluated both gross skin findings as well as histopathological findings. Using gross skin findings alone resulted in an accuracy of 85.7% which could be increased to 100% accuracy when the gross skin findings were combined with histopathological findings. This study suggests that the inclusion of histopathological findings increases the accuracy of the phototoxicity evaluation of systemically administered drugs in SD rats. In conclusion, we found that for studying drug-induced phytotoxicity, a 10 J/cm(2) UVA dose serves as the optimal radiation dose, and that the inclusion of histopathological findings increases the accuracy of the phototoxicity evaluation of the drugs.

Human skins are exposed to nanomaterials in everyday life from various sources such as nanomaterial-containing cosmetics, air pollutions, and industrial nanomaterials. Nanomaterials comprising metal haptens raises concerns about the skin sensitization to nanomaterials. In this study, we evaluated the skin sensitization of nanomaterials comparing metal haptens in vivo and in vitro. We selected five metal oxide NPs, containing copper oxide, cobalt monoxide, cobalt oxide, nickel oxide, or titanium oxide, and two types of metal chlorides (CoCl2 and CuCl2), to compare the skin sensitization abilities between NPs and the constituent metals. The materials were applied to KeratinoSensTM cells for imitated skin-environment setting, and luciferase induction and cytotoxicity were evaluated at 48 h post-incubation. In addition, the response of metal oxide NPs was confirmed in lymph node of BALB/C mice via an in vivo method. The results showed that CuO and CoO NPs induce a similar pattern of positive luciferase induction and cytotoxicity compared to the respective metal chlorides; Co3O4, NiO, and TiO2 induced no such response. Collectively, the results implied fast-dissolving metal oxide (CuO and CoO) NPs release their metal ion, inducing skin sensitization. However, further investigations are required to elucidate the mechanism underlying NP-induced skin sensitization. Based on ion chelation data, metal ion release was confirmed as the major “factor” for skin sensitization.

High crystalline ZnO nanorods (NRs) on Zn pre-deposited graphene/Cu sheet without graphene transfer process have been fabricated by self-catalyzed vapor-phase transport synthesis. Here, the pre-deposited Zn metal on graphene not only serves as a seed to grow the ZnO NRs, but also passivates the graphene underneath. The temperature-dependent photoluminescence spectra of the fabricated ZnO NRs reveal a dominant peak of 3.88 eV at 10 K associated with the neutral-donor bound exciton, while the redshifted peak by bandgap shrinkage with temperature and electron-lattice interactions leads a strong emission at 382 nm at room temperature. The optical absorption of the ZnO NRs/graphene hetero-nanostructure at this ultraviolet (UV) emission is then theoretically analyzed to quantify the absorption amount depending on the ZnO NR distribution. By simply covering the ZnO NR/graphene/Cu structure with the graphene/glass as a top electrode, it is observed that the current-voltage characteristic of the ZnO NR/graphene hetero-nanojunction device exhibits a photocurrent of 1.03 mA at 3 V under a light illumination of 100 μW/cm2. In particular, the suggested graphene/ZnO NRs/graphene hybrid-nanostructure-based devices reveal comparable photocurrents at a bidirectional bias, which can be a promising platform to integrate 1D and 2D nanomaterials without complex patterning process for UV device applications.

A spectrophotometric method based on diazo-coupling reaction for nitrite analysis was established and validated, including inter-laboratory validation, linearity, accuracy, precision, the limit of detection (LOD) and limit of quantification (LOQ). The time-saving and high-recovery method was established by examining the filtration step, colorimetric process and concentration range of the calibration curve. This method showed good linearity (r2 > 0.999) in the range of 0.025–1.0 μg/mL. The three-level recoveries were between 86.7% and 108.6%, with the coefficient of variation (CV) below 5.8%. Mean nitrite concentration ranges in processed foods were ND–33.47 mg/kg. The mean nitrite intake was 0.8% of the Acceptable Daily Intake (ADI, 0.07 mg/kg bw/day) for all children and adolescents and 2.8% for the consumer group. The major contributors for all subjects and consumers were ham, sausage and bacon. These results indicated that the improved method was suitable for analyzing nitrite in processed foods and the nitrite exposure levels were safe.

Some drugs cause phototoxicity in humans when exposed to light, thus there is a need for an in vivo phototoxicity test to evaluate them. However, an in vivo phototoxicity test method to evaluate this has not been established. This study aimed to establish an in vivo phototoxicity test method for transdermally administered drugs. For this, we evaluated the phototoxicity using Sprague-Dawley (SD) rats for transdermal administered drugs and we studied the appropriate UVA dose using 8-methoxypsalen, which is a well-known phototoxic drug. We found that a UVA dose of 15 J/cm2 was dose and time dependent response compared to other UVA doses. We performed the Minimum Erythema Dose (MED) test because UVB can cause skin irritation by itself and selected 0.01 J/cm2 as an appropriate dose of UVB. Using the selected UVA and UVB doses, we performed a phototoxicity study of 6 pharmaceutical drugs, which included phototoxic and non-phototoxic drugs. As a result of the phototoxicity test, 100% accuracy was obtained when compared with previous studies. In addition, we performed histopathology to confirm the new findings. We found that histopathology can be used as an additional indicator of phototoxicity test for transdermally administered drugs.

The compound 6:2 chlorinated polyfluorinated ether sulfonate (F-53B), a replacement for perfluorooctanesulfonate (PFOS) in the electroplating industry, has been widely detected in numerous environmental matrices, human sera, and organisms. Due to regulations that limit PFOS use, F-53B use is expected to increase. Therefore, in this study, we performed a subchronic oral toxicity study of F-53B in Sprague Dawley (SD) rats. F-53B was administered orally once daily to male and female rats for 28 days at doses of 5, 20, and 100 mg/kg/day. There were no toxicologically significant changes in F-53B-treated rats, except in the thyroid gland. However, F-53B slightly reduced the serum concentrations of thyroid hormones, including triiodothyronine and thyroxine, compared with their concentrations in the vehicle group. F-53B also induced follicular hyperplasia and was associated with increased thyroid hormone biosynthesis-associated protein expression. These results demonstrate that F-53B is a strong regulator of thyroid hormones in SD rats as it disrupts thyroid function. Thus, caution should be exercised in the industrial application of F-53B as an alternative for PFOS.

Numerous studies have reported the potential of chemicals for inducing skin sensitization; however, few studies have examined skin sensitization induced by nanomaterials. This study aimed to evaluate skin sensitization induced by metal oxide nanoparticles (NPs) using the ARE-Nrf2 Luciferase KeratinoSens™ assay. Seven different metal oxide NPs, including copper oxide, cobalt oxide, nickel oxide, titanium oxide, cerium oxide, iron oxide, and zinc oxide, were assessed on KeratinoSens™ cells. We selected an appropriate vehicle among three vehicles (DMSO, DW, and culture medium) by assessing the hydrodynamic size at vehicle selection process. Seven metal oxide NPs were analyzed, and their physicochemical properties, including hydrodynamic size, polydispersity, and zeta potential, were determined in the selected vehicle. Thereafter, we assessed the sensitization potential of the NPs using the ARE-Nrf2 Luciferase KeratinoSens™ assay. Copper oxide NPs induced a positive response, whereas cobalt oxide, nickel oxide, titanium oxide, cerium oxide, iron oxide, and zinc oxide NPs induced no response. These results suggest that the ARE-Nrf2 Luciferase KeratinoSens™ assay may be useful for evaluating the potential for skin sensitization induced by metal oxide NPs.

Although skin sensitization potential of various chemicals has been extensively studied, there are only a few reports on nanoparticles induced skin sensitization. Aiming to fill this lacuna, in this study we evaluated the potential of metal oxide nanoparticles (NPs) to induce skin sensitization with flow cytometry. Seven different metal oxide NPs, including copper oxide, cobalt oxide, nickel oxide, titanium oxide, cerium oxide, iron oxide, and zinc oxide were applied to Balb/c mice. After selecting the proper vehicle, the NPs were applied, and the skin sensitization potential were assessed using 5-bromo-2-deoxyuridine with flow cytometry. Physiochemical properties such as hydrodynamic size, polydispersity, and zeta potential were measured for the NPs prior to the tests. All the seven metal oxide NPs studied showed negative responses for skin sensitization potential. These results suggest that the OECD TG 442B using 5-bromo-2-deoxyuridine with flow cytometry can be applied to evaluate the potential of NPs for skin sensitization.

The mutagenic potencies of 1,3-propane sultone (PS), N-propyl-N-nitrosourea (PNU), and mitomycin C (MMC) were investigated in three independent laboratories in Korea using the Pig-a assay in vivo. Sprague-Dawley rats were treated with vehicle or test substance on three consecutive days. Blood samples were collected for measuring Pig-a mutant phenotypes (CD59-deficient erythrocytes, RBCCD59-; CD59-deficient reticulocytes, RETCD59-) on days -1, 15, and 29 after the first treatment. In some studies, blood was collected for determining DNA damage (comet assay) on day 3 and measuring micronucleated reticulocytes (MN-RET) on day 4. Treatment with the alkylating agents PS and PNU induced dose-dependent increases in the frequency of RBCCD59- on days 15 and 29, and caused maximum elevations in the frequency of RETCD59- on day 15. Inter-laboratory comparison of the day 29 Pig-a assay data confirmed the mutagenic potencies of PS and PNU, and showed good agreement among the test sites. Treatment with the DNA cross-linker MMC induced increases in the frequencies of RBCCD59- and RETCD59- on days 15 and 29 (all three laboratories). MN-RETs increased significantly in animals treated with PS, PNU, or MMC, but biologically significant increases in DNA damage were observed only with PS and PNU, and not with MMC. The results of this study indicate that the Pig-a assay is a sensitive, reproducible method for evaluating the in vivo mutagenicity of various test substances, in particular, DNA cross-linkers and alkylating agents. Our limited data on integrating the Pig-a assay with the comet and micronucleus assays indicate that a short-term treatment protocol evaluating these three endpoints in a single set of animals may be a robust strategy for evaluating in vivo genotoxicity.

This study performed an experiment on the causes of pitting corrosion in a copper tubing used for a sprinkler system. Corrosion products of a copper tubing that sustained pitting corrosion were collected and cultured in Culture medium [Luria–Bertani, Brain heart infusion, Tryptic soy broth (TSB), R2A]. Four types of bacteria were found through identification: Micrococcus luteus sp (species)., Staphylococcus sp., Sphingomonas sp., and Bacillus sp. The copper toxicity test was performed for each microorganism. Among the four microorganisms, Micrococcus luteus sp. showed good growth in the environment containing copper ions. On the immersion test, changes in pH and Optical density were measured; On the inductively coupled plasma optical emission spectrometry test, the copper concentration of each culture medium was measured. The surface of each copper sample was observed using a scanning electron microscope. The corrosion potential of a copper sample, after 48 h exposure of the TSB medium containing Micrococcus luteus sp., was measured using a potentiodynamic polarization experiment. The next experiment was conducted to prevent microbial corrosion by suppressing the growth of microorganisms. Six 30 ml TSB culture media with controlled pH value of 4, 5, 6, 7, 8, and 9 through HCl and NaOH were manufactured. Then the microorganisms were cultured in 37 °C 133 rpm, of which the growth status was checked every 24 h for 3 days. It was found that microorganisms did not grow on culture media with the pH value of 6 and lower. The same experiment conducted on culture media controlled with acetic acid, nitric acid, and sulfuric acid, also showed no growth of microorganisms on media with pH value of 6 and lower. Six 5 ml TSB culture media each containing 0.5, 0.25, 0.125, 0.0625, 0.0312%, and 0.0156% NaOCl and NaOBr as germicides were manufactured. 0.01 μl of microorganisms were inoculated on the media and cultured in 37 °C for 48 h. It was found that microorganisms did not grow in media with NaOCl and NaOBr concentration of 0.0625% and higher. Therefore, it can be suggested that in environments with pH value of 6 or lower, or NaOCl and NaOBr concentration of higher than 0.0625% suppresses microbial growth, thereby preventing microbial corrosion.

We demonstrate a controllable and reliable process for manifesting color patterns on solid substrates using cellulose nanocrystals (CNCs) without the use of any other chemical pigments. The color can be controlled by adjusting the assembly conditions of the CNC solution during a dip-and-pull process while aiding the close packing of CNCs on a solid surface with the help of ionic-liquid (1-butyl-3-methylimidazolium) molecules that screen the repelling electrostatic charges between CNCs. By controlling the pulling speed from 3 to 9 μm/min during the dip-and-pull process, we were able to control the film thickness from 100 to 300 nm, resulting in films with different colors in the visible range. The optical properties were in good agreement with the finite-difference time-domain simulation results. By functionalizing these films with amine groups, we developed colorimetric sensors that can change in color when exposed to aldehyde gases such as formaldehyde or propanal. A principal component analysis showed that we can differentiate between different aldehyde gases and other interfering molecules. We expect that our approach will enable inexpensive and rapid volatile organic compound detection with on-site monitoring capabilities.

Evaluation of DNA damage is critical during the development of new drugs because it is closely associated with genotoxicity and carcinogenicity. The in vivo comet assay to assess DNA damage is globally harmonized as OECD TG 489. However, a comet test guideline that evaluates DNA damage without sacrificing animals does not yet exist. The goal of this study was to select an appropriate cell line for optimization of the in vitro comet assay to assess DNA damage. We then evaluated the predictivity of the in vitro comet assay using the selected cell line. In addition, the effect of adding S9 was evaluated using 12 test chemicals. For cell line selection, HepG2, Chinese hamster lung (CHL/IU), and TK6 cell lines were evaluated. We employed a method for the in vitro comet assay based on that for the in vivo comet assay. The most appropriate cell line was determined by% tail DNA increase after performing in vitro comet assays with 6 test chemicals. The predictivity of the in vitro comet assay using the selected cell line was measured with 10 test chemicals (8 genotoxins and 2 non-genotoxic chemicals). The HepG2 cell line was found to be the most appropriate, and in vitro comet assays using HepG2 cells exhibited a high accuracy of 90% (9/10). This study suggests that HepG2 is an optimal cell line for the in vitro comet assay to assess DNA damage.

Zinc oxide nanoparticles (ZnO NPs) are used in various industries such as food additives, cosmetics, and biomedical applications. In this study, we evaluated lung damage over time by three types of ZnO NPs (L-serine, citrate, and pristine) following the regulation of functional groups after a single intratracheal instillation to rats. The three types of ZnO NPs showed an acute inflammatory reaction with increased LDH and inflammatory cell infiltration in the alveoli 24 h after administration. Especially in treatment with L-serine, citrate ZnO NPs showed higher acute granulocytic inflammation and total protein induction than the pristine ZnO NPs at 24 h. The acute inflammatory reaction of the lungs recovered on day 30 with bronchoalveolar fibrosis. The concentrations of IL-4, 6, TNF-α, and eotaxin in the bronchoalveolar lavage fluid (BALF) decreased over time, and the levels of these inflammation indicators are consistent with the following inflammatory cell data and acute lung inflammation by ZnO NP. This study suggests that single inhalation exposure to functionalized ZnO NPs may cause acute lung injury with granulocytic inflammation. Although it can recover 30 days after exposure, acute pulmonary inflammation in surface functionalization means that additional studies of exposure limits are needed to protect the workers that produce it.

Skin sensitisation is the regulatory endpoint that has been at the centre of concerted efforts to replace animal testing in recent years, as demonstrated by the Organisation for Economic Co-operation and Development (OECD) adoption of five non-animal methods addressing mechanisms under the first three key events of the skin sensitisation adverse outcome pathway. Nevertheless, the currently adopted methods, when used in isolation, are not sufficient to fulfil regulatory requirements on the skin sensitisation potential and potency of chemicals comparable to that provided by the regulatory animal tests. For this reason, a number of defined approaches integrating data from these methods with other relevant information have been proposed and documented by the OECD. With the aim to further enhance regulatory consideration and adoption of defined approaches, the European Union Reference Laboratory for Alternatives to Animal testing in collaboration with the International Cooperation on Alternative Test Methods hosted, on 4-5 October 2016, a workshop on the international regulatory applicability and acceptance of alternative non-animal approaches, i.e., defined approaches, to skin sensitisation assessment of chemicals used in a variety of sectors. The workshop convened representatives from more than 20 regulatory authorities from the European Union, United States, Canada, Japan, South Korea, Brazil and China. There was a general consensus among the workshop participants that to maximise global regulatory acceptance of data generated with defined approaches, international harmonisation and standardisation are needed. Potential assessment criteria were defined for a systematic evaluation of existing defined approaches that would facilitate their translation into international standards, e.g., into a performance-based Test Guideline. Informed by the discussions at the workshop, the ICATM members propose practical ways to further promote the regulatory use and facilitate adoption of defined approaches for skin sensitisation assessments.

Introduction Neuromyelitis optica (NMO) is a rare inflammatory autoimmune disorder of the CNS. Rituximab is used to treat antibody-mediated autoimmune diseases. Case presentation We report the case a patient with NMO, who was treated with rituximab and presented CD20+ T cells by flow cytometry after treatment, later diagnosed with lung B-cell lymphoma. Conclusion This is the first report of CD20+ T cell detection in an NMO patient. We found that CD20+ T cells recovered faster than B cells after rituximab treatment and that CD20+ T cells seemed to play a role in suppressing tumor growth and memory T cell activity.

Although alternative test methods based on the 3Rs (Replacement, Reduction, Refinement) are being developed to replace animal testing in reproductive and developmental toxicology, they are still in an early stage. Consequently, we aimed to develop alternative test methods in male animals using mouse spermatogonial stem cells (mSSCs). Here, we modified the OECD TG 489 and optimized the in vitro comet assay in our previous study. This study aimed to verify the validity of in vitro tests involving mSSCs by comparing their results with those of in vivo tests using C57BL/6 mice by gavage. We selected hydroxyurea (HU), which is known to chemically induce male reproductive toxicity. The 50% inhibitory concentration (IC50) value of HU was 0.9 mM, as determined by the MTT assay. In the in vitro comet assay, % tail DNA and Olive tail moment (OTM) after HU administration increased significantly, compared to the control. Annexin V, PI staining and TUNEL assays showed that HU caused apoptosis in mSSCs. In order to compare in vitro tests with in vivo tests, the same substances were administered to male C57BL/6 mice. Reproductive toxicity was observed at 25, 50, 100, and 200 mg/kg/day as measured by clinical measures of reduction in sperm motility and testicular weight. The comet assay, DCFH-DA assay, H&E staining, and TUNEL assay were also performed. The results of the test with C57BL/6 mice were similar to those with mSSCs for HU treatment. Finally, linear regression analysis showed a strong positive correlation between results of in vitro tests and those of in vivo. In conclusion, the present study is the first to demonstrate the effect of HU-induced DNA damage, ROS formation, and apoptosis in mSSCs. Further, the results of the current study suggest that mSSCs could be a useful model to predict male reproductive toxicity.