Multi-Sensor Data Fusion with a Reconfigurable Module and Its Application to Unmanned Storage Boxes.

We present a multi-sensor data fusion model based on a reconfigurable module (RM) with three fusion layers. In the data layer, raw data are refined with respect to the sensor characteristics and then converted into logical values. In the feature layer, a fusion tree is configured, and the values of the intermediate nodes are calculated by applying predefined logical operations, which are adjustable. In the decision layer, a final decision is made by computing the value of the root according to predetermined equations. In this way, with given threshold values or sensor characteristics for data refinement and logic expressions for feature extraction and decision making, we reconstruct an RM that performs multi-sensor fusion and is adaptable for a dedicated application. We attempted to verify its feasibility by applying the proposed RM to an actual application. Considering the spread of the COVID-19 pandemic, an unmanned storage box was selected as our application target. Four types of sensors were used to determine the state of the door and the status of the existence of an item inside it. We implemented a prototype system that monitored the unmanned storage boxes by configuring the RM according to the proposed method. It was confirmed that a system built with only low-cost sensors can identify the states more reliably through multi-sensor data fusion.

Multiwall Carbon Nanotube-Induced Apoptosis and Antioxidant Gene Expression in the Gills, Liver, and Intestine of Oryzias latipes.

Multiwall carbon nanotubes (MWCNTs) have many attractive properties with potential applications in various fields. Despite their usefulness, however, the associated waste can be hazardous to the environment. To examine adverse effects in aquatic environments, Oryzias latipes were exposed to MWCNTs dispersed in water for 14 days and apoptosis and antioxidant gene expression were observed. This work showed that in gills exposed to 100 mg/L MWCNTs for 4 days, there was significant p53, caspase-3 (Cas3), caspase-8 (Cas8), and caspase-9 (Cas9) gene expression relative to the controls, while catalase (CAT) and glutathione-S-transferase (GST) expression were reduced. At 14 days, CAT, GST, and metallothionein (MT) were induced significantly in the gills and Cas3, Cas8, and Cas9 were induced in the liver. No significant gene induction was seen in intestine. Intracellular reactive oxygen species (ROS) were increased significantly only at 14 days. Histologically, no apoptosis was observed with exposure to 100 mg/L MWCNTs for 21 days. The gills were more sensitive to MWCNT toxicity than the other organs. Males had higher apoptosis gene induction than females. These results demonstrated that MWCNTs could cause apoptosis in a manner influenced by tissue and gender in aqueous environments.

Cloning metallothionein gene in Zacco platypus and its potential as an exposure biomarker against cadmium.

Zacco platypus, pale chub, is an indigenous freshwater fish of East Asia including Korea and has many useful characteristics as indicator species for water pollution. While utility of Z. platypus as an experimental species has been recognized, genetic-level information is very limited and warrants extensive research. Metallothionein (MT) is widely used and well-known biomarker for heavy metal exposure in many experimental species. In the present study, we cloned MT in Z. platypus and evaluated its utility as a biomarker for metal exposure. For this purpose, we sequenced complete complementary DNA (cDNA) of MT in Z. platypus and carried out phylogenetic analysis with its sequences. The transcription-level responses of MT gene following the exposure to CdCl2 were also assessed to validate the utility of this gene as an exposure biomarker. Analysis of cDNA sequence of MT gene demonstrated high conformity with those of other fish. MT messenger RNA (mRNA) expression and enzymatic MT content significantly increased following CdCl2 exposure in a concentration-dependent manner. The level of CdCl2 that resulted in significant MT changes in Z. platypus was within the range that was reported from other fish. The MT gene of Z. platypus sequenced in the present study can be used as a useful biomarker for heavy metal exposure in the aquatic environment of Korea and other countries where this freshwater fish species represents the ecosystem.

Benzo(a)pyrene-induced cytochrome p4501A expression of four freshwater fishes (Oryzias latipes, Danio rerio, Cyprinus carpio, and Zacco platypus).

Oryzias latipes, Danio rerio, Cyprinus carpio, and Zacco platypus are useful indicator species for CYP1A biomarker studies; however, comparative studies have not been performed. To compare susceptibility, dose- and time-dependent CYP1A induction at the mRNA and protein levels in response to benzo(a)pyrene (BaP) exposure was analyzed. At the mRNA level, a statistically significant difference was found among the four species; however, such was not observed at the protein level. C. carpio showed the highest CYP1A induction level and the steepest slope in the dose-response curve. To assess susceptibility, the difference in CYP1A mRNA induction among species must be considered, and C. carpio was the most sensitive species of the four evaluated in terms of CYP1A expression.

Tri-Metal Layered Semitransparent Electrode for Red Phosphorescent Organic Light-Emitting Diodes.

In this paper, we fabricated tri-metal layered thin film semitransparent electrodes consisting of a thin conductive metal layer, sandwiched between two nickel layers. An equal red phosphorescent organic light-emitting diode (PHOLED) structure was deposited on the anodes of indium tin oxide (ITO) and three types of tri-metal layers (Ni/Al/Ni, Ni/Cu/Ni, and Ni/Ag/Ni, thickness of 3/7/3 nm in common) on a glass substrate. The optical and electrical performances of the device using Ni/Ag/Ni were improved more than the performances of the other devices due to the micro-cavity effect in accordance with the various electrode characteristics. Moreover, we fabricated the same red PHOLED structures on a flexible substrate, as a consequence, showed competitive emission characteristics compared to the devices fabricated on a glass substrate. Therefore, this study could succeed to additional research on flexible display panel and light-emitting devices with ITO-free electrodes.

Integration of multi-level biomarker responses to cadmium and benzo[k]fluoranthene in the pale chub (Zacco platypus).

The Cd exposure for 14 days significantly increased both the molecular (DNA single-strand breaks) and biochemical (metallothionein concentrations) biomarkers in the freshwater pale chub, Zacco platypus, whereas changes in the histological and physiological biomarker responses were negligible. The BkF exposure for 14 days led to significant increases in the mRNA expression of catalase and superoxide dismutase, 7-ethoxyresorufin-O-deethylase enzymatic activity and DNA single-strand breakage at the molecular and biochemical levels. In addition, exposure to 50µg/L of BkF induced histological alteration in the liver, with significant changes to the liver somatic index and condition factor at the physiological level. The integration of multi-level biomarker responses at the molecular, biochemical and physiological levels was highly correlated with the concentrations of Cd and BkF.

Uptake, tissue distribution, and depuration of total silver in common carp (Cyprinus carpio) after aqueous exposure to silver nanoparticles.

The increased use and disposal of silver nanoparticles (AgNPs) has led to their release from wastewater treatment plants into surface waters and concern over potential for negative effects in aquatic organisms. Investigations of the toxicity of AgNPs in fish have considered various species, exposure routes, and test end points; however, the toxicokinetics of total silver has not been studied in fish exposed to aqueous AgNPs. In this study, we investigated the toxicokinetics of total silver in common carp (Cayprinus carpio) exposed to AgNPs [0.62 ± 0.12 (mean ± standard deviation) mg L(-1)] for 7 days followed by a 2 week depuration period. During exposure and depuration, fish were sampled, tissues were excised (gills, brain, skeletal muscle, gastrointestinal tract, liver, and blood) and digested in acid, and total silver concentrations were analyzed by inductively coupled plasma-optical emission spectrometry. Total silver in tissues increased during the 7 day exposure, and mean concentrations were 5.61 mg/kg of liver, 3.32 mg/kg of gills, 2.93 mg/kg of gastrointestinal tract, 0.48 mg/kg of skeletal muscle, 0.14 mg/kg of brain, and 0.02 mg/kg of blood. Transmission electron microscopy energy-dispersive spectroscopy confirmed the presence of silver in the tissues. After depuration for 14 days, total silver returned to control levels in all tissues except liver (4.22 mg/kg), gastrointestinal tract (1.26 mg/kg), and gills (0.77 mg/kg).

Toxicity of citrate-coated silver nanoparticles differs according to method of suspension preparation.

To evaluate substance toxicity, it is critical to maintain specific concentrations of test substances throughout the exposure period. During the last decade, the need to improve methods for nanoparticle (NP) suspension preparations has gained attention because many published results on NPs toxicity have been inconsistent. Here, we compared the toxicity of citrate-coated silver nanoparticles (AgNPs) suspended by two different methods (fractionated vs. colloidal) in freshwater organisms (daphnia and medaka). Analytical methods (ICP-OES, DLS and UV absorbance) were employed to characterize behavior of AgNPs in suspension. Results showed that fractionated (stirred and settled) solution was less toxic to daphnia (13.8 µg/L) than colloidal solution (6.1 µg/L), suggesting that method of preparation was a critical factor that affected toxicity. However, differences in toxicity caused by suspension methods were not observed in medaka. Results indicate that the method used to prepare suspensions of NPs can affect toxicity, and that differences can exist among test organisms.

Tissue-specific antioxidant responses in pale chub (Zacco platypus) exposed to copper and benzo[a]pyrene.

In this study, antioxidant responses including lipid peroxidation (LPO), superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST), were evaluated in the liver, gill and muscle tissues of pale chub (Zacco platypus) exposed to copper (Cu) and benzo[a]pyrene (BaP). Cu exposure induced significant antioxidant responses in Z. platypus, particularly in the liver, whereas BaP exposure had a negligible effect. Following Cu exposure, both SOD and CAT activity increased in a concentration-dependent manner, showing significant correlations with malondialdehyde (MDA) levels as a measure of LPO (r = 0.646 and 0.663, respectively). SOD, CAT and GST mRNA levels were also enhanced following Cu exposure, except at 20 µg L(-1), although significant correlations with antioxidant enzyme activities were not found. The results of this study suggest that combined information on SOD and CAT activities together with LPO levels in the liver could be a useful indicator for assessing oxidative stress in freshwater fish.

TopBP1 deficiency impairs V(D)J recombination during lymphocyte development.

TopBP1 was initially identified as a topoisomerase II-ß-binding protein and it plays roles in DNA replication and repair. We found that TopBP1 is expressed at high levels in lymphoid tissues and is essential for early lymphocyte development. Specific abrogation of TopBP1 expression resulted in transitional blocks during early lymphocyte development. These defects were, in major part, due to aberrant V(D)J rearrangements in pro-B cells, double-negative and double-positive thymocytes. We also show that TopBP1 was located at sites of V(D)J rearrangement. In TopBP1-deficient cells, γ-H2AX foci were found to be increased. In addition, greater amount of γ-H2AX product was precipitated from the regions where TopBP1 was localized than from controls, indicating that TopBP1 deficiency results in inefficient DNA double-strand break repair. The developmental defects were rescued by introducing functional TCR αß transgenes. Our data demonstrate a novel role for TopBP1 as a crucial factor in V(D)J rearrangement during the development of B, T and iNKT cells.

Cytochrome P450 system expression and DNA adduct formation in the liver of Zacco platypus following waterborne benzo(a)pyrene exposure: implications for biomarker determination.

Benzo(a)pyrene (BaP) is a polycyclic aromatic hydrocarbon that causes mutations and tumor formation. Zacco platypus is a sentinel species that is suitable for monitoring aquatic environments. We studied cytochrome P450 system (CYP system) expression and DNA adduct formation in the liver of Z. platypus following waterborne exposure to BaP. The results showed both dose and time dependency. The significant induction levels of CYP system mRNA and protein reached maximums at 2 days and 14 days, respectively, and hepatosomatic index was maximally induced at 4 days during 14 days BaP exposure. DNA adduct formation was significantly induced compared to corresponding controls (t-test, p < 0.01) after 4 days of exposure in 100 µg/L BaP. These results indicate that the only use of mRNA expression level of CYP system as a biomarker make us underestimate prolonged toxicity (4-14 days) of BaP and the only use of protein expression level of CYP system make us underestimate acute toxicity (1-2 days) of BaP. Therefore, we suggests that a combinational use of the mRNA expression level and protein expression level of CYP system, hepatosomatic index is a useful biomarker in risk assessment of waterborne BaP exposure. In addition, DNA adduct formation was a useful biomarker in risk assessment of waterborne BaP exposure at 4 days. CYP1A was a more sensitive biomarker than CYP reductase for BaP exposure when considering both the mRNA and protein level. Furthermore, our results show that Z. platypus is a useful species for assessing the risk of waterborne BaP exposure.

Effect of material properties on stability of silver nanoparticles in water.

Increased production and use of silver nanoparticles (AgNPs) could potentially lead to their release into the environment. Estimating the exposure to engineered nanomaterials in the environment is essential for assessing their risk. This study examined the aggregation and sedimentation kinetics behaviors of citrate- (Cit-AgNPs) and polyvinylpyrrolidone-coated silver nanoparticles (PVP-AgNPs) with three different average sizes in calcium chloride (CaCl2) solutions, emphasizing the effects of particle size and type of coating material on both behaviors. As the ionic strength increased, Cit-AgNPs (stabilized by charge repulsion) aggregated rapidly and settled down, while PVP-AgNPs (stabilized by steric repulsion) did not aggregate, even at an ionic strength of 10 mM CaCl2, due to likely steric hiderance effects of PVP coating. Interestingly, however, PVP-AgNPs sedimented without aggregating within 7 days and this tendency seems to having relevance to the particle size. These results suggest that the particle size and type of coating material play important roles in determining nanoparticle fate and transport.

Proteomic analysis in Daphnia magna exposed to As(III), As(V) and Cd heavy metals and their binary mixtures for screening potential biomarkers.

In this study, the effects of three widespread heavy metals, As(III), As(V) and Cd, and their binary mixtures on the proteomic profile in D. magna were examined to screen novel protein biomarkers using the two-dimensional gel electrophoresis method (2DE). Ten 20d daphnia were exposed to the LC20 concentrations for each of a total of 8 treatments, including the control, As(III), As(V), Cd, [As(III)+As(V)], [As(III)+Cd], [As(V)+Cd], and [As(III), As(V), Cd], for 24h before protein isolation. Three replicates were performed for each treatment. These protein samples were employed for 2DE experiments with a pH gradient gel strip from pH 3 to pH 10. The protein spots were detected by a silver staining process and their intensities were analyzed by Progenesis software to discover the differentially expressed proteins (DEPs) in response to each heavy metal. A total of 117 differentially expressed proteins (DEPs) were found in daphnia responding to the 8 treatments and mapped onto a 2D proteome map, which provides some information of the molecular weight (MW) and pI value for each protein. All of these DEPs are considered as potential candidates for protein biomarkers in D. magna for detecting heavy metals in the aquatic ecosystem. Comparing the proteomic results among these treatments suggested that exposing D. magna to binary mixtures of heavy metals may result in some complex interactive molecular responses within them, rather than just the simple sum of the proteomic profiles of the individual chemicals, (As(III), As(V), and Cd).

Effects of water chemistry on aggregation and soil adsorption of silver nanoparticles.

OBJECTIVES: In this study, we investigated the influence of ionic strength and natural organic matter (NOM) on aggregation and soil adsorption of citrate-coated silver nanoparticles (AgNPs). METHODS: Time-resolved dynamic light scattering measurements and batch adsorption experiments were used to study their aggregation and soil adsorption behaviors, respectively. RESULTS: The aggregation rate of AgNPs increased with increasing ionic strength and decreasing NOM concentration. At higher ionic strength, the AgNPs were unstable, and thus tended to be adsorbed to the soil, while increased NOM concentration hindered soil adsorption. To understand the varying behaviors of AgNPs depending on the environmental factors, particle zeta potentials were also measured as a function of ionic strength and NOM concentration. The magnitude of particle zeta potential became more negative with decreasing ionic strength and increasing NOM concentration. These results imply that the aggregation and soil adsorption behavior of AgNPs were mainly controlled by electrical double-layer repulsion consistent with the Derjaguin-Landau-Verwey-Overbeek theory. CONCLUSIONS: This study found that the aggregation and soil adsorption behavior of AgNPs are closely associated with environmental factors such as ionic strength and NOM and suggested that assessing the environmental fate and transport of nanoparticles requires a thorough understanding of particle-particle interaction mechanisms.

Integrative assessment of biomarker responses in pale chub (Zacco platypus) exposed to copper and benzo[a]pyrene.

Multi-level biomarker responses (molecular/biochemical and histological/physiological levels) were studied to assess the sublethal toxicities of copper (Cu: 1.25, 5, and 20µg/L) and benzo[a]pyrene (BaP: 0.5, 5, and 50µg/L) induced in the freshwater pale chub Zacco platypus. Except for the kidney tissues when exposed to 20µg Cu/L, no significant differences were observed at the histological or physiological levels among the treatment groups. However, various molecular and biochemical responses were observed in Z. platypus, and these responses primarily depended on exposure time. Upon Cu exposure, both DNA single-strand breaks (COMET) and metallothionein (MT) concentration significantly increased after 4 days, whereas there were no significant changes after 14 days of exposure. Both 4 and 14 days of BaP exposure induced significant increases in COMET and 7-ethoxyresorufin-O-deethylase (EROD) activity, but there was no significant difference between them. Additionally, both Cu and BaP induced acetylcholinesterase (AChE) activity only after 14 days of exposure. The current findings demonstrate that the differences in the responses of MT and EROD are associated with each chemical's particular mode of action. Biomarker responses at the molecular and biochemical levels were quantized in terms of the integrated biomarker response (IBR) index to compare the toxicities of Cu and BaP. The IBR values were well correlated with the concentrations of Cu and BaP, and the correlations were enhanced at 4 days of exposure (r(2)=0.849 and 0.945, respectively) compared with 14 days (r(2)=0. 412 and 0.634, respectively). These results suggest that the IBR index may be a useful tool for the integrative quantification of the molecular and biochemical biomarker responses in a short-term exposure to Cu and BaP.

Development of an Enzyme-linked Immunosorbent Assay Using Vitellin for Vitellogenin Measurement in the Pale Chub, Zacco platypus.

OBJECTIVES: Fish vitellogenin (VTG) is produced in the female liver during oogenesis through the estradiol cycle and produced in the male liver by endocrine disrupting chemicals (EDCs) such as alkylphenols. In this study, we propose that the VTG concentration in the pale chub could be detected using monoclonal antibodies and polyclonal antibodies against vitellin (Vn) in a VTG enzyme-linked immunosorbent assay (ELISA) system. METHODS: Monoclonal antibodies and polyclonal antibodies were produced using the Vn extracted from the matured ovum of the ovary. The VTG was extracted from the plasma of the male pale chub. The Vn and VTG were confirmed by measuring the molecular weight of their proteins using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the specificity of the antibodies was checked through western blotting methods. The assay system was validated with respect to optimal assay concentrations, specificity, recovery, and intra- and inter-assay variations. RESULTS: The Vn consisted of two protein bands with apparent molecular weights of 64 and 37 kDa. The SDS-PAGE indicated protein weights of 146 and 77 kDa in the VTG. The assay range was 15.6 ng/mL to 2,000 ng/mL, and the value of the intra- and inter-assay variations were within 10.0% and 14.7%, respectively. The recovery rate was 99.5±5.5%. CONCLUSIONS: A sandwich ELISA was developed that could be used to qualify the VTG of pale chub in screening for EDCs. Pale chub is an ideal species for observing estrogen activity in the environment because of its extensive habitat and extensive food chain. The ELISA developed here would be more favorable than those for other species for determining the effect of long-term food chain accumulation of EDCs in aquatic environments.

A novel function of adipocytes in lipid antigen presentation to iNKT cells.

Systemic low-grade chronic inflammation has been intensively investigated in obese subjects. Recently, various immune cell types, such as macrophages, granulocytes, helper T cells, cytotoxic T cells, and B cells, have been implicated in the pathogenesis of adipose tissue inflammation. However, the roles of invariant natural killer T cells (iNKT cells) and the regulation of iNKT cell activity in adipose tissue are not thoroughly understood. Here, we demonstrated that iNKT cells were decreased in number in the adipose tissue of obese subjects. Interestingly, CD1d, a molecule involved in lipid antigen presentation to iNKT cells, was highly expressed in adipocytes, and CD1d-expressing adipocytes stimulated iNKT cell activity through physical interaction. iNKT cell population and CD1d expression were reduced in the adipose tissue of obese mice and humans compared to those of lean subjects. Moreover, iNKT cell-deficient Jα18 knockout mice became more obese and exhibited increased adipose tissue inflammation at the early stage of obesity. These data suggest that adipocytes regulate iNKT cell activity via CD1d and that the interaction between adipocytes and iNKT cells may modulate adipose tissue inflammation in obesity.

Toxicity Assessment of Titanium (IV) Oxide Nanoparticles Using Daphnia magna (Water Flea).

OBJECTIVES: Titanium dioxide (TiO(2)), a common nanoparticle widely used in industrial production, is one of nano-sized materials. The purpose of this study was to determine the acute and chronic toxicity of TiO(2) using different size and various concentrations on Daphnia magna. METHODS: In the acute toxicity test, four concentrations (0, 0.5, 4, and 8 mM) for TiO(2) with 250 or 500 nm and five concentrations (0, 0.25, 0.5, 0.75, and 1 mM) for TiO(2) with 21 nm were selected to analyze the toxic effect to three groups of ten daphnia neonates over 96 hours. In addition, to better understand their toxicity, chronic toxicity was examined over 21 days using 0, 1, and 10 mM for each type of TiO(2). RESULTS: Our results showed that all organisms died before the reproduction time at a concentration of 10 mM of TiO(2). In addition, the exposure of anatase (21 nm) particles were more toxic to D. magna, comparing with that of anatase (250 nm) and rutile (500 nm) particles. CONCLUSIONS: This study indicated that TiO(2) had adverse impacts on the survival, growth and reproduction of D. magna after the 21days exposure. In addition, the number of test organisms that were able to reproduce neonates gradually were reduced as the size of TiO(2) tested was decreased.

Silver nanoparticles induce apoptosis and G2/M arrest via PKCζ-dependent signaling in A549 lung cells.

The use of silver nanoparticles is one of the fastest growing product categories in the nanotechnology industry, with a focus on antimicrobial activity. However, thus far, toxicity data for silver nanoparticles are limited. In this study, we investigated the cytotoxic effects of silver nanoparticles (Ag NPs) and the pathway by which they affect A549 lung epithelial cells. The effects of Ag NPs on cell survival, cell cycle progression, and mRNA and protein alterations of selected cell cycle- and apoptosis-related genes were studied using formazan dye and LDH release assays, flow cytometric analysis, semi-quantitative RT-PCR, and Western blot analysis. Ag NPs reduced cell viability, increased LDH release, and modulated cell cycle distribution through the accumulation of cells at G2/M and sub-G1 phases (cell death), with a concurrent decrease in cells at G1. Ag NP treatment increased Bax and Bid mRNA levels and downregulated Bcl-2 and Bcl-w mRNAs in a dose-dependent manner. Furthermore, Ag NPs altered the mRNA levels of protein kinase C (PKC) family members. In particular, ectopic overexpression of PKCζ led to the enhancement of cellular proliferation and reduced sensitivity to Ag NPs in A549 cells. Together, these results suggest that Ag NPs induce strong toxicity and G2/M cell cycle arrest by a mechanism involving PKCζ downregulation in A549 cells.

Toxicity evaluation of verapamil and tramadol based on toxicity assay and expression patterns of Dhb, Vtg, Arnt, CYP4, and CYP314 in Daphnia magna.

In this study, the toxicities of two pharmaceuticals, verapamil and tramadol were evaluated in Daphnia magna using the conventional toxicity tests (acute and chronic test) and the expression patterns of five stress responsive genes. In the chronic toxicity test, several parameters, such as the survival percentage, the body length of D. magna, the time of first reproduction, and the number of offspring per female, were adversely affected during the exposure to 4.2 mg L(-1) verapamil and 34 mg L(-1) tramadol. During the 24-h short-term exposure, verapamil particularly caused a downregulated expression of the CYP4 and CYP314 genes, whereas tramadol upregulated the expression of the CYP314 gene. Neither pharmaceutical affected the expression of Dhb, Arnt, and Vtg. However, during the 21-day long-term exposure, both verapamil and tramadol significantly reduced the expression level of the Vtg gene, a biomarker of the reproduction ability in an oviparous animal, whereas neither affected the other genes.