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.

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.

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.

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).

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.

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.

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.

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.

The SWI/SNF chromatin-remodeling complex modulates peripheral T cell activation and proliferation by controlling AP-1 expression.

The SWI/SNF chromatin-remodeling complex has been implicated in the activation and proliferation of T cells. After T cell receptor signaling, the SWI/SNF complex rapidly associates with chromatin and controls gene expression in T cells. However, the process by which the SWI/SNF complex regulates peripheral T cell activation has not been elucidated. In this study, we show that the SWI/SNF complex regulates cytokine production and proliferation of T cells. During T cell activation, the SWI/SNF complex is recruited to the promoter of the transcription factor AP-1, and it increases the expression of AP-1. Increased expression of the SWI/SNF complex resulted in enhanced AP-1 activity, cytokine production, and proliferation of peripheral T cells, whereas knockdown of the SWI/SNF complex expression impaired the AP-1 expression and reduced the activation and proliferation of T cells. Moreover, mice that constitutively expressed the SWI/SNF complex in T cells were much more susceptible to experimentally induced autoimmune encephalomyelitis than the normal mice were. These results suggest that the SWI/SNF complex plays a critical role during T cell activation and subsequent immune responses.

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.

Analysis of the effects diclofenac has on Japanese medaka (Oryzias latipes) using real-time PCR.

The expression levels of cytochrome P450 1A, p53 and vitellogenin were investigated in three different tissues of male medaka fish after exposure to diclofenac that is one of the main concerns among pharmaceuticals frequently found in sewage treatment plant (STP) effluents. The results showed that cytochrome P450 1A, p53 and vitellogenin were highly expressed in tissue-specific gene expression patterns after exposure to 8 mg/l and 1 microg/l of diclofenac. These elevated expression levels of three biomarkers suggested that diclofenac has potential to cause cellular toxicity, p53-related genotoxicity and estrogenic effects. It is also noteworthy that diclofenac has the potential to cause these effects even at an environmentally relevant concentration of diclofenac, 1 microg/l.

Stressor identification and health assessment of fish exposed to wastewater effluents in Miho Stream, South Korea.

This study evaluated the effects of an industrial wastewater treatment plant (IWTP) and a municipal wastewater treatment plant (MWTP) effluents on a variety of bioindicators ranging from biochemical, organism, and population-level responses in pale chub (Zacco platypus) and fish community structure. The Index of Biotic Integrity (IBI) indicated that the site upstream of these wastewater treatment plant discharges is in fair-good condition and downstream of the plant is in poor condition. The EROD (ethoxyresorufin-O-deethylase) activity, condition factor, and liver somatic index were significantly increased at the downstream site compared to those of the upstream site. The most significant change observed in pale chub population in the downstream site of the Miho Stream, relative to the upstream population, was the total absence of an younger age group. Stressors impacting the downstream site were identified as mostly organic or nutrient enrichment and habitat degradation associated with wastewater treatment plants. The results of causal analysis suggest that the primary causes affecting fish population in the downstream site are through both size-selective mortality caused by ammonia toxicity and recruitment failure caused by habitat degradation and reproduction problem due to an IWTP and MWTP effluents.

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.

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.

Inhibition of oocyte development in Japanese medaka (Oryzias latipes) exposed to di-2-ethylhexyl phthalate.

Di-2-ethylhexyl phthalate (DEHP) is a widely used plasticizer known to be a suspected endocrine disrupter, but its precise effects on aquatic organisms are not yet known. When Japanese medaka (Oryzias latipes) were exposed from the time of hatching to 3 months of age to an aqueous DEHP solution at nominal concentrations of 1, 10, and 50 microg/l, DEHP-treated female fish showed distinct reproductive effects as follows. First, blood vitellogenin levels in all treated test subjects markedly decreased. Second, Gonado Somatic Index (GSI) decreased to 33% and 38% of the control GSI in 10 microg/l and 50 microg/l treated female fish, respectively. Third, 54% of female fish in the control treatment had completely matured oocytes in their ovaries, but only 37%, 0% and 22% of female fish matured to the last stage in the 1, 10 and 50 microg/l treated test subjects, respectively. Unlike female fish, no change or adverse effects were observed in the male fish. In summary, DEHP hinders the development of reproductive organs in the female Japanese medaka. In this work, the possibility o f anti-estrogenic activity of DEHP is proposed as the cause.

Pattern recognition of the movement tracks of medaka (Oryzias latipes) in response to sub-lethal treatments of an insecticide by using artificial neural networks.

Specimens of medaka (Oryzias latipes) were observed continuously through an automatic image recognition system before and after treatments of an anti-cholinesterase insecticide, diazinon (0.1 mg/l), for 4 days in semi-natural conditions (2 days before treatment and 2 days after treatment). The "smooth" pattern was typically shown as a normal movement behavior, while the "shaking" pattern was frequently observed after treatments of diazinon. These smooth and shaking patterns were selected for training with an artificial neural network. Parameters characterizing the movement tracks, such as speed, degree of backward movements, stop duration, turning rate, meander, and maximum distance movements in the y-axis of 1-min duration, were given as input (six nodes) to a multi-layer perceptron with the back propagation algorithm. Binary information for the smooth and shaking patterns was separately given as the matching output (one node), while eight nodes were assigned to a single hidden layer. As new input data were given to the trained network, it was possible to recognize the smooth and shaking patterns of the new input data. Average recognition rates of the smooth pattern decreased significantly while those for the shaking pattern increased to a higher degree after treatments of diazinon. The trained network was able to reveal the difference in the shaking pattern in different light phases before treatments of diazinon. This study demonstrated that artificial neural networks could be useful for detecting the presence of toxic chemicals in the environment by serving as in-situ behavioral monitoring tools.

Histologic changes of pulmonary arteries in congenital heart disease with left-to-right shunt (part 1): correlated with preoperative pulmonary hemodynamics. Emphasizing the significance of pulmonary arterial concentration.

The hemodynamic findings related to the histologic changes in the small pulmonary arteries in congenital heart disease were analyzed with a left-to-right shunt. A lung biopsy was performed during the repair because of pulmonary arterial (PA) hypertension (mean PA pressure > or = 15 mmHg) in 38 patients. There were 13 patients whose age ranged from 2 years to 25 years old. A preoperative cardiac catheterization was performed to locate the site of the defect and to determine the preoperative hemodynamic findings. Among them, 29 patients had pulmonary vascular resistance (PVR) > 2.5 unit/m2. The lung biopsy specimens were investigated microscopically for the Heath- Edward grade, morphometric analysis of medial wall thickness (MWT) and the rate of the decrease in the pulmonary arteriolar concentration (PAC) obtained by the alvoelo-arterial ratio divided by patients' age. All patients were in the Heath-Edward grade I to III (29 patients in grade I). This grade correlated with the MWT, but did not correlate with a decrease in the PAC. The MWT and the rate of the decrease in the PAC did not accompany each other, but either one had the tendency dominate the pattern in individual patients. The MWT had a close correlation with the mean PA pressure and PVR, and an even closer correlation in patients with a high PVR and those older than 2 years of age. The rate of the decreased in the PAC showed a weak correlation with the shunt volume in patients over 2 years of age or with a large shunt. In the high flow group (PVR < 2.5 unit/m(2), Qp/Qs > 2.0, n=14) the MWT was significantly thinner and the rate of the decrease in the PAC was significantly higher than the high resistance group (PVR > 2.5 unit/m2, Qp/Qs2 < 2.0, n=13). The rate of the decrease in the PAC correlated with the patients' age, but the MWT did not. The lung biopsy results in patients who had both left-to-right shunts and pulmonary hypertension showed that the rate of the decrease in the PAC was weakly related to the shunt volume and the MWT was related to the PA pressure and PVR. Either an increased MWT or the rate of the decrease in the PAC tended to dominate. These phenomena were prominent in patients older than 2 in whom a wide range of individual variations were noted in the morphometric pattern. The medial hypertrophy and the rate of the decrease in the PAC may be induced by different stimuli or that medial hypertrophy may play a role in preventing PAC decrease.