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The tetraspanin superfamily proteins are transmembrane proteins identified in a diverse range of eukaryotic organisms. Tetraspanins are involved in a variety of essential biological functions, including cell differentiation, adhesion, migration, signal transduction, intracellular trafficking, and immune responses. For an infection to occur, viruses must interact with various cell surface components, including receptors and signaling molecules. Tetraspanin CD63 is involved in the organization of the cell membrane and trafficking of cellular transmembrane proteins that interact with many viruses. In this study, the cd63 gene was characterized by studying its expression and function in a zebrafish model. The functional domains and structural features of Cd63, such as the Cys-Cys-Gly (CCG) motif in the large extracellular loop and cysteine residues, are conserved in zebrafish. We confirmed that cd63 was expressed in immune system organs, such as the axial vein and pronephric duct, during the embryonic development of zebrafish. To better understand the role of cd63 in the zebrafish immune system, we established cd63 -deficient zebrafish lines using the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) system. A 19 bp insertion mutation was generated in single guide RNA (sgRNA) target sequence of exon 3 of the cd63 gene, to create a pre-mature stop codon. We then analyzed the expression of cd63 -related genes cxcr4a and cxcr4b in wild type (WT) and cd63 -deficient zebrafish. We believe our study provides an important model that could be used to investigate the roles of cd63 in viral infection in vivo. • Characterization of cd63 from zebrafish (Danio rerio). • Expression analysis at the developmental stage of zebrafish by in situ hybridization. • The cd63 expression was significantly increased upon the VHSV infection. • Cd63 knockout zebrafish was generated using a CRISPR/Cas9 system. • Cd63 -related genes, cxcr4a and cxcr4b , were downregulated in cd63 knockout zebrafish. [ABSTRACT FROM AUTHOR]

In this study, we used three processes to synthesize magnesium sulfate from seawater. First, alkaline industrial wastes, cement kiln dust and paper sludge ash, were injected to the seawater to precipitate magnesium in the form of magnesium hydroxide (Mg(OH)2). Then, magnesium was eluted with a small amount of H2SO4 to make a high concentration magnesium solution. Finally, an organic solvent was added to precipitate magnesium sulfate (MgSO4). Over 90% of magnesium was recovered through the three processes. It is expected that 11.3 kg of magnesium sulfate (based on MgSO4 · 6H2O) can be synthesized from 1 ton of seawater. [ABSTRACT FROM AUTHOR]

Abstract Inflammasomes promote the production of pro-inflammatory cytokines, such as interleukin (IL)-1β and IL-18, which are the representative mediators of inflammation. Abnormal activation of inflammasomes leads to the development of inflammatory diseases such as acute pancreatitis (AP). In this study, we demonstrate the inhibitory effects of a new natural compound fraxinellone on inflammasome formation and examine the role of inflammasomes in a mouse model of AP. AP was induced with hourly intraperitoneal injections of supramaximal concentrations of the stable cholecystokinin analogue cerulein (50 μg/kg) for 6 h. Mice were sacrificed 6 h after the final cerulein injection. Blood and pancreas samples were obtained for further experiments. Intraperitoneal injection of fraxinellone significantly inhibited the pancreatic activation of multiple inflammasome molecules such as NACHT, LRR and PYD domains-containing protein 3 (NLRP3), PY-CARD, caspase-1, IL-18, and IL-1β during AP. In addition, fraxinellone treatment inhibited pancreatic injury, elevation in serum amylase and lipase activities, and infiltration of inflammatory cells such as neutrophils and macrophages but had no effect on pancreatic edema. To investigate whether inflammasome activation leads to the infiltration of inflammatory cells, we used parthenolide, a well-known natural inhibitor, and IL-1 receptor antagonist mice. The inhibition of inflammasome activation by pharmacological/or genetic modification restricted the infiltration of inflammatory cells, but not edema, consistent with the results observed with fraxinellone. Taken together, our study highlights fraxinellone as a natural inhibitor of inflammasomes and that inflammasome inhibition may lead to the suppression of inflammatory cells during AP. Graphical abstract Unlabelled Image Highlights • Fraxinellone is a natural inhibitor of inflammasome. • Inflammasome leads to the infiltration of inflammatory cells into pancreas during acute pancreatitis. • Fraxinellone inhibits pancreatic injury via suppression of inflammasome against acute pancreatitis. [ABSTRACT FROM AUTHOR]

• A high-performance hydrogen generator was developed using solid-state NaBH 4. • The satisfying performance of hydrogen supply was obtained for various flight conditions. • High purity of hydrogen of 99.64% was obtained for the entire conditions. • Performance for dynamic response to sudden hydrogen demand was evaluated. A solid NaBH 4 -based hydrogen generator was developed as a hydrogen source for fuel-cell-powered unmanned autonomous systems (UAS). Although many studies have been conducted to extract hydrogen from solid NaBH 4 , performance evaluation required for commercialization are still insufficient so that a new structure of the hydrogen generator is required to meet the performance requirement. For this purpose, in this study, a new reactor was made of a magnesium alloy with perfluoroalkoxy coating to improve its thermal conductivity and chemical resistance. In addition, a porous partition was installed inside the reactor to separate the hydrogen buffer volume from the reaction zone where NaBH 4 granules was filled. Next, The operational characteristics of the developed hydrogen generator were evaluated in high- and low-temperature environments, inclined pitch and roll angles, unpredictable power cut-off situations, and sudden hydrogen demand, including the idle, rated, and maximum power modes. A start-up of the hydrogen generator was possible without additional heating and insulation at −15 °C and the hydrogen generation was stable after start-up for all pitch (±30°) and roll (±45°) angles, whereas the CoG shifted forward by 26% of the total length as the agent solution was consumed. In addition, the hydrogen supply immediately recovered within 30 s after an unpredictable power cut-off. Hydrogen purity was 99.64% on average and impurities originated from the water vapor. Finally, the dynamic response to the sudden hydrogen demand and precise controllability of the hydrogen supply rate were tested according to the simulated hydrogen consumption profile for practical use in fuel cells. In conclusion, the developed hydrogen generator satisfied the requirements for commercialization. [ABSTRACT FROM AUTHOR]

• Two ferritin like subunits (LhFerH & LhFerM) were identified from Liza haematocheila. • Recombinant ferritin subunits showed detectable iron deprivation activity. • Both recombinant subunits could inhibit the growth of E.coli in LB media. • Both LhFerH and LhFerM expressed universality in all selected fish tissues. • In-vivo expression of both LhFerH and M was induced post immune stimulation. Iron (Fe) is considered as an essential micronutrient due to its diverse functions in living systems. However, regulation of free iron levels is essential because free Fe ions, in excess, induce biological toxicity through different routes, including production of reactive oxygen species. Ferritin proteins play a vital role in controlling free Fe ion homeostasis by sequestering excess iron in the body. Ferritins comprise an H subunit with a ferroxidase center and an L subunit with a Fe nucleation site. However, lower vertebrates such as fish harbor an additional subunit termed ferritin M, which shows the characteristic features of both H and L. In this study, two ferritin subunits (H and M) with ferroxidase centers were identified and characterized from red-lip mullet (Liza haematocheila). The open reading frames of red-lip mullet ferritin H (LhFerH) and ferritin M−like (LhFerM) subunits comprise 534 and 531 bps, which encode for putative polypeptides of 177 and 176 amino acids, respectively. LhFerH and LhFerM were found to retain well-conserved residues, including seven ferroxidase di-iron centers, characteristic domains, and signatures of their known homologs. We cloned the open reading frames of the two ferritin subunits to overexpress the corresponding proteins in Escherichia coli and subsequently demonstrated their iron sequestration activity along with antibacterial activity against E. coli using respective purified recombinant proteins in vitro. A basal expression analysis of two LhFer genes in selected tissues using qPCR assays showed pronounced expression in blood cells with respect to both genes. A relatively high expression level of LhFerH was also detected in muscle tissues. The expression level of LhFer in the head kidney was significantly up-regulated following lipopolysaccharides (LPS) and Lactococcus garvieae injection. The resulting gene expression pattern upon immune stimulation suggests that ferritin may contribute to the defense against harmful pathogen infection. Collectively, our results indicate that both LhFerH and LhFerM potentially participate in the homeostasis of free Fe ions and in the host immune defense of red-lip mullet. [ABSTRACT FROM AUTHOR]

Spring viremia of carp (SVC) is listed as a notifiable viral disease by the World Organization for Animal Health (OIE). In 2016, the first official SVC outbreak was detected in the city of Gyeongsan, Korea. The present study reports the first complete genome analysis of SVC virus (SVCV, ADC-SVC2016-5) isolated from leather carp (Cyprinus carpio nudus). The results revealed that ADC-SVC2016-5 has a 11,029-bp genome containing five genes: N, P, M, G, and L. Phylogenetic analysis indicated that ADC-SVC2016-5 (accession number MG663512), isolated from leather carp, was closely related to genogroup Ia isolates of the Asian clade. This report provides additional information for studying the molecular epidemiology and evolution of spring viremia of carp virus. [ABSTRACT FROM AUTHOR]

Acute pancreatitis (AP) is an inflammatory disease of the pancreas, which, in its most severe form, is associated with multi-organ failure and death. Loganin, a major iridoid glycoside obtained from Corni fructus , has been shown to have anti-inflammatory and anti-shock effects. However, the effects of loganin on AP have not been determined. Pre-treatment of loganin reduced pancreatic damage and AP-associated lung injury and attenuated the severity of AP, as evidenced by (1) a reduction in several biochemical parameters (pancreatic weight to body weight ratio, myeloperoxidase activity, and level of amylase) and (2) production of pro-inflammatory cytokines such as interleukin (IL)-1β and tumor necrosis factor (TNF)-α. However, post-treatment of loganin failed to improve pancreatic damage and biochemical parameters of AP, but could inhibit the AP-induced elevation of IL-1β and TNF-α significantly. In addition, cerulein-induced activation of nuclear factor (NF)-κB was inhibited in the pancreas by administration of loganin. In conclusion, these results suggest that loganin exhibits an anti-inflammatory effect in cases of AP and its pulmonary complications through inhibition of NF-κB activation. [ABSTRACT FROM AUTHOR]

Stimulator of interferon genes (STING) is an adapter protein that is activated when cyclic dinucleotides (CDNs) are present. CDNs originate from the cytosolic DNA of both pathogens and hosts. STING activation promotes efficient immune responses against viral infections; however, its impact in bacterial infections is unclear. In this study, we investigated the role of Sting in bacterial infections by successfully creating a sting-deficient (sting(−/−) with a 4-bp deletion) knockout zebrafish model using CRISPR/Cas9. The transcriptional modulation of genes downstream of cGAS (cyclic GMP-AMP synthase)-Sting pathway-related genes was analyzed in seven-day-old wild-type (WT) and sting(−/−) embryos, as well as in four-day-old LPS-stimulated embryos. The expression of downstream genes was higher in sting(−/−) than in healthy WT fish. The late response was observed in sting(−/−) larvae following LPS treatment, demonstrating the importance of Sting-induced immunity during bacterial infection by activating the cGAS–STING pathway. Furthermore, adult sting(−/−) fish had a high mortality rate and significantly downregulated cGAS–STING pathway-related genes during Edwardsiella piscicida (E. piscicida) infection. In addition, we assessed NF-κB pathway genes following E. piscicida infection. Our results show fluctuating patterns of interleukin-6 (il6) and tumor necrosis factor-α (tnfα) expression, which is likely due to the influence of other NF-κB pathway-related immune genes. In summary, this study demonstrates the important role of Sting against bacterial infection. [ABSTRACT FROM AUTHOR]

In this study, we developed a technology to recover Mg from seawater desalination brine in the form of high-purity MgSO 4 that did not contain Ca impurities. The MgSO 4 recovery consisted of pre-precipitation of Mg(OH) 2 using alkali, concentration of Mg using sulfuric acid, and precipitation of MgSO 4 using ethanol. The removal of Ca impurities by adding ethanol twice into the Mg concentrate in the precipitation process was subtle but a highly beneficial novelty. The first dosage of ethanol was added to the Mg concentrate by adjusting the quantity such that only Ca (without Mg) was precipitated in the form of CaSO 4. Then, the ethanol was further added to the Ca-free Mg concentrate to precipitate high-purity MgSO 4. This two-step ethanol addition ensures effective removal of Ca impurities without Mg loss using the difference in the solubility of CaSO 4 and MgSO 4 in ethanol. As the purity of the recovered MgSO 4 was up to 99.8%, it could be used to re-mineralize fresh water after the seawater desalination process. According to the cost assessment, high-purity MgSO 4 produced from seawater desalination brine is expected to be preferred over other products in pharmaceutical and food markets that require high purity and economic feasibility. • We develop a technology to produce pure MgSO 4 from seawater desalination brine. • Ca impurities are effectively removed without Mg loss by adding ethanol twice. • Purity of Ca-free MgSO 4 produced is very high up to 99.8%. • The technology uses the solubility difference between CaSO 4 and MgSO 4 in ethanol. • Mg recovery efficiency is 67%, producing 15.8 kg of MgSO 4 ·7H 2 O from 1 ton of brine. [ABSTRACT FROM AUTHOR]

Abstract: This study aims at evaluating the reuse feasibility of effluent produced by the soil washing of mine tailings with oxalic acid. Alkaline chemicals such as NaOH, Ca(OH)2, and Na2CO3 are used for the precipitation of arsenic and heavy metals in the effluent containing oxalic acid. All of the target contaminants are removed with very high efficiency (up to 100%) at high pH. The precipitation using NaOH at pH 9 is determined to be the most cost-effective method for the removal of arsenic as well as heavy metals in the effluent. The effluent decontaminated by NaOH is consecutively reused for the soil washing of raw mine tailings, resulting in considerable efficiency. Furthermore, even more arsenic and heavy metals are extracted from raw mine tailings by acidifying the decontaminated effluent under the alkaline condition, compared with direct reuse of the decontaminated effluent. Here, the oxalic acid, which is a weak complex-forming ligand as well as a weak acid, has noticeable effects on both soil washing and effluent treatment by precipitation. It extracts efficiently the contaminants from the mine tailings without adverse change of soil and also makes possible the precipitation of the contaminants in the effluent unlike strong chelating reagent. Reuse of the washing effluent containing oxalic acid would make the existing soil washing process more environment-friendly and cost-effective. [Copyright &y& Elsevier]

In this paper, we propose a fast rate control algorithm in frame-layer of H.264/AVC video coding standard. We derive a statistical relation between quantization parameter (QP) and encoded bits of inter-coded blocks. In addition, new complexity estimation is defined to determine weight of complexity of current frame. Using the statistical property and the complexity updating function, we determine the amount of bits as a function of quantization parameter, so that computational cost of rate control is significantly reduced with coding gain improvement and stable buffer management. Experimental results demonstrate the capability of the proposed algorithm. [ABSTRACT FROM PUBLISHER]

Abstract: During vertebrate heart valve formation, Wnt/β-catenin signaling induces BMP signals in atrioventricular canal (AVC) myocardial cells and underlying AVC endocardial cells then undergo endothelial-mesenchymal transdifferentiation (EMT) by receiving this BMP signals. Histone deacetylases (HDACs) have been implicated in numerous developmental processes by regulating gene expression. However, their specific roles in controlling heart valve development are largely unexplored. To investigate the role of HDACs in vertebrate heart valve formation, we treated zebrafish embryos with trichostatin A (TSA), an inhibitor of class I and II HDACs, from 36 to 48h post-fertilization (hpf) during which heart looping and valve formation occur. Following TSA treatment, abnormal linear heart tube development was observed. In these embryos, expression of AVC myocardial bmp4 and AVC endocardial notch1b genes was markedly reduced with subsequent failure of EMT in the AVC endocardial cells. However, LiCl-mediated activation of Wnt/β-catenin signaling was able to rescue defective heart tube formation, bmp4 and notch1b expression, and EMT in the AVC region. Taken together, our results demonstrated that HDAC activity plays a pivotal role in vertebrate heart tube formation by activating Wnt/β-catenin signaling which induces bmp4 expression in AVC myocardial cells. [Copyright &y& Elsevier]

In this study, the soil washing technique has been used to treat mine tailings contaminated heavily with arsenic and heavy metals at Jingok mine, which is one of the abandoned mines in Korea. The results showed that phosphoric acid, citric acid, oxalic acid, and sodium metabisulfite were highly effective in extracting arsenic and heavy metals. Among them, oxalic acid was the most effective (especially for Pb, Cu, and Zn), as even a residual fraction of arsenic was partly extracted. The optimum concentration of washing reagent and the ratio between the mine tailings and washing reagent were found to be 0.5 M and 1:20, respectively. In addition, the extraction kinetics of arsenic and heavy metals was fast, in which the reaction time of 30 minutes was deemed to be a sufficient contact time. From the results, it may be concluded that the low pH of washing solution and the amount of dissolved Fe may be considered as the most important factors in the extraction of arsenic and heavy metals. [ABSTRACT FROM AUTHOR]

Monte Carlo simulations have been used extensively in studying the performance of control charts. Researchers have used various numbers of replications in their studies, but almost none of them provided justifications for the number of replications used. Currently, there are no empirically based recommendations regarding the required number of replications to ensure accurate results. This research examined six recently published studies to develop recommendations for the minimum number of replications necessary to reproduce the reported results within a specified degree of accuracy. The results of this study indicated that using 10,000 replications was unnecessarily large and a smaller number of replications could be used to reproduce the target ARLs within the 2% error bands satisfying the modified Mundfrom's criteria. In many cases, only 5,000 replications or fewer were required. In general, the number of replications required to reproduce the target ARL decreased as the shift size increased. In addition, the results of this study provide general recommendations for the required number of replications to use in future SPC simulation studies. [ABSTRACT FROM AUTHOR]

In the present paper, inorganic arsenic species and chemical parameters in groundwater were determined to investigate the factors related to the distribution of arsenic species and their dissolution from rock into groundwater. For the study, groundwater and core samples were taken at different depths of two newly drilled wells in Huron and Lapeer Counties, Michigan. Results show that total arsenic concentrations in the core samples varied, ranging from 0.8 to 70.7 mg/kg. Iron concentration in rock was about 1800 times higher than that of arsenic, and there was no correlation between arsenic and iron occurrences in the rock samples. Arsenic concentrations in groundwater ranged from <1 to 171 μg/l. The arsenic concentration in groundwater depended on the amount of arsenic in aquifer rocks, and as well decreased with increasing depth. Over 90% of arsenic existed in the form of As(III), implying that the groundwater systems were in the reduced condition. The results such as high ferrous ion, low redox potential and low dissolved oxygen supported the observed arsenic species distribution. There was no noticeable difference in the total arsenic concentration and arsenic species ratio between unfiltered and filtered (0.45 μm) waters, indicating that the particulate form of arsenic was negligible in the groundwater samples. There were correlations between water sampling depth and chemical parameters, and between arsenic concentration and chemical parameters, however, the trends were not always consistent in both wells. [Copyright &y& Elsevier]

Previous studies on the utilization of seawater and brine have focused on either recovering Mg from seawater/brine using alkalis or extracting Ca from industrial byproducts using solvents. Therefore, the present study was aimed at recovering Mg and Ca simultaneously from simulated seawater/brine and slaked lime [Ca(OH) 2 ] using a simple process, with seawater/brine employed both as a Mg source and as a solvent for extracting Ca from the Ca source. Manipulating the concentration of the sucrose additive and the ionic strength of seawater/brine enabled Mg and Ca recovery with high efficiency and purity. The highest Mg recovery from brine was 75 %, and ~ 100 % pure MgSO 4 was produced; specifically, ~28 kg of MgSO 4 •7H 2 O was produced from 1 ton of brine. The impacts of sucrose concentration and seawater ionic strength on the Ca elution, carbonation efficiency, and size and shape of the resulting vaterite particles were analyzed. The purity of the vaterite produced was 94 %–100 %, and the maximum CaCO 3 yield per ton of the Ca(OH) 2 was estimated as 770 kg, with a carbonation efficiency of up to 92 %. Therefore, our cost-effective method—featuring the use of seawater/brine and Ca(OH) 2 —can concurrently produce economically demanding minerals such as MgSO 4 and vaterite, thereby facilitating resource conservation. • MgSO 4 and CaCO 3 are produced simultaneously from seawater and Ca(OH) 2. • The sucrose content and seawater ionic strength affect the mineral yield and form. • The Mg recovery efficiency is 75 %, and the MgSO 4 •7H 2 O yield is 28 kg/ton of brine. • The carbonation efficiency is 92 %, and the CaCO 3 yield is 770 kg/ton of Ca(OH) 2. • The MgSO 4 and vaterite powders obtained through the devised process are ~100 % pure. [ABSTRACT FROM AUTHOR]

• Seawater and industrial by-product are used for CO 2 storage and CaCO 3 production. • Economic feasibility of the indirect carbonation is secured by nearly costless seawater. • Mg in seawater mainly contributes to eluting Ca from alkali industrial by-product. • CO 2 storage and CaCO 3 yield are 271 and 615 kg/ton-CKD, respectively. • Purity of CaCO 3 exceeds 99% despite the use of CKD and seawater with impurities. Indirect carbonation is one of the typical carbon capture, utilization, and storage technologies. It is well known, however, that the technology is very difficult to achieve economic feasibility because expensive chemical solvents used account for most of the cost. To overcome this limitation, we performed an experimental study to secure the economic feasibility of the technology by replacing such chemical solvents with nearly costless seawater. For the study, we used cement kiln dust (CKD), which is an alkali industrial by-product, together with seawater. In this paper, we attained CO 2 storage and CaCO 3 yield despite the use of seawater, which is comparable in both quantitative and qualitative respects to the existing studies using chemical reagents. The CO 2 storage and CaCO 3 yield were 185 kg-CO 2 /ton-CKD and 419 kg-CaCO 3 /ton-CKD, respectively. With the addition of Mg into the seawater, moreover, the amounts could significantly increase to reach 271 kg-CO 2 /ton-CKD and 615 kg-CaCO 3 /ton-CKD, respectively. Despite using CKD and seawater containing many impurities, the purity of CaCO 3 produced was as high as 99.4%. It was also found that Mg is one component, which can elute Ca from CKD, dissolved in seawater. The solid to liquid ratio was the most influential factor for the Ca elution efficiency, while the CO 2 flow rate and NaOH dosage had significant effects on the carbonation efficiency. [ABSTRACT FROM AUTHOR]

• High-density hydrogen generator was developed for unmanned aerial vehicles. • Solid-state NaBH 4 was used as a hydrogen source for high storage density. • Stable hydrogen generation was obtained for the entire operation time. • Restartability was evaluated to validate the possibility of commercialization. A hydrogen generator using solid-state sodium borohydride (NaBH 4) as a hydrogen source was proposed as part of a high-energy-density fuel-cell system for unmanned aerial vehicles. In this study, solid-state NaBH 4 was used in order to overcome problems of a hydrogen generator using catalytic hydrolysis of NaBH 4 aqueous solution such as low hydrogen yield and unstable hydrogen generation due to catalyst degradation. An agent solution, hydrochloric acid, was injected on to the NaBH 4 to generate hydrogen. In addition, all of the balance-of-plant systems and components were integrated to develop a lightweight, commercially-viable, high-density hydrogen generator. The developed hydrogen generator was evaluated in terms of stable hydrogen generation and restartability for the entire operation time in order to validate the possibility of commercialization. From the performance evaluation, the gravimetric and volumetric specific energy densities of the hydrogen generator were found to be 739.1 W hr/kg and 272.8 W hr/L, respectively. In addition, the hydrogen storage density was 5.1 wt%, which was 1.44 times higher in comparison of a typical hydrogen generator using NaBH 4 aqueous solution. [ABSTRACT FROM AUTHOR]

Chronic pancreatitis (CP) is characterized by recurrent pancreatic injury, resulting in inflammation and fibrosis. Currently, there are no drugs for the treatment of pancreatic fibrosis associated with CP. Piperine, a natural alkaloid found in black pepper, has been reported to show anti-inflammatory, anti-oxidative, and antitumor activities. Although piperine exhibits numerous properties in regards to the regulation of diverse diseases, the effects of piperine on CP have not been established. To investigate the effects of piperine on CP in vivo, we induced CP in mice through the repetitive administration of cerulein (50 µg/kg) six times at 1-h intervals, 5 times per week, for a total of 3 weeks. In the pre-treatment groups, piperine (1, 5, or 10 mg/kg) or corn oil were administrated orally at 1 h before the first cerulein injection, once a day, 5 times a week, for a total of 3 weeks. In the post-treatment groups, piperine (10 mg/kg) or corn oil was administered orally at 1 or 2 week after the first cerulein injection. Pancreases were collected for histological analysis. In addition, pancreatic stellate cells (PSCs) were isolated to examine the anti-fibrogenic effects and regulatory mechanisms of piperine. Piperine treatment significantly inhibited histological damage in the pancreas, increased the pancreatic acinar cell survival, reduced collagen deposition and reduced pro-inflammatory cytokines and chemokines. In addition, piperine treatment reduced the expression of fibrotic mediators, such as α-smooth muscle actin (α-SMA), collagen, and fibronectin 1 in the pancreas and PSCs. Moreover, piperine treatment reduced the production of transforming growth factor (TGF)-β in the pancreas and PSCs. Furthermore, piperine treatment inhibited TGF-β-induced pSMAD2/3 activation but not pSMAD1/5 in the PSCs. These findings suggest that piperine treatment ameliorates pancreatic fibrosis by inhibiting TGF-β/SMAD2/3 signaling during CP. [ABSTRACT FROM AUTHOR]

[Display omitted] • Nanosized 100 % vaterite was produced via organic-solvent-free indirect carbonation. • Seawater, sucrose, ultrasonication, and aging exhibited a synergistic effect. • An eluent with the highest free-Ca-ion concentration was prepared by controlling the sucrose concentration. • The carbonation end pH was optimized, and the stirring speed and ultrasound intensity were adjusted to minimize interference. • After carbonation, the CaCO 3 -containing suspension was floated and a stable dispersion was maintained. Nanosized vaterite, which exhibits characteristics such as high specific surface area, porosity, and biocompatibility, has attracted research attention for use as a drug delivery material. However, fatal drawbacks such as high costs, difficulty in mass production, and toxicity exist in conventional nanosized vaterite production owing to the use of a large amount of organic solvents to forcibly suppress the vaterite recrystallization and particle growth. Therefore, nanosized 100 % vaterite was produced in this study via indirect carbonation without using any organic solvent, which has rarely been achieved previously. Seawater, sucrose, ultrasonication, and aging—which facilitate vaterite production and particle size reduction—exhibited a synergistic effect in producing vaterite. To realize nanosized vaterite production via indirect carbonation, seawater was used as a solvent, sucrose was added when Ca was eluted, and CO 2 bubbling was performed under ultrasonication. Furthermore, the CaCO 3 -containing suspension obtained after the carbonation was aged. Ultrasonic waves were required to generate nanosized vaterite and reducing size at the carbonation stage. This nanosized-vaterite-production strategy involving organic-solvent-free indirect carbonation is meaningful, in that it highlights the potential of synthesizing vaterite in an economically sound, environmentally friendly manner for use as a pharmaceutical raw material. [ABSTRACT FROM AUTHOR]

Furfuryl alcohol, which is the hydrogenated product of furfural, has been identified as a very promising platform chemical with high potential for applications in the manufacture of key chemicals, lubricants, fragrances, and pharmaceuticals. In this work, bare SB, and x % Ru/Al‐SB (x=1.5, 2.5, 3.5, and 4.5 wt. %) samples were fabricated by a hydrothermal method. Bare and most active catalysts were characterized by different techniques, such as BET, FE‐SEM, TEM, FT‐IR, and XRD, to understand their physical and chemical properties. An evaluation of the effects of various reaction parameters, such as catalyst loading, reaction temperature, and reaction time, on the catalytic performance, showed higher catalytic conversion of furfural and selectivity for the desired products. The most active RuS3 catalyst showed 100 % conversion of furfural and 99 % selectivity for furfuryl alcohol. It could be reused for five consecutive reaction cycles without significant loss of performance. In addition, Ru leaching and loss of conversion or selectivity were not noticed during the five‐run recycling test. The EDS elemental mapping analysis of the used catalyst established the preservation of the mesoporous structure, suggesting a strong interaction between the hexagonal porous silicate and the Ru nanoparticles. [ABSTRACT FROM AUTHOR]

Dynamic Voltage and Frequency Scaling (DVFS) is an effective low-power technique for real-time workloads. Its effectiveness critically depends on the accurate prediction of the task execution time. Many DVFS approaches have been proposed, but they are insufficient for highly nonstationary workloads. Several recent DVFS techniques adopted adaptive filters to improve accuracy. However, their improvement was rather limited, since they mainly focused on applying a filter framework to the target application without tuning it. We address this issue by proposing Particle Filter (PF)-based video decoders (MPEG2 and H.264) which exploit application-specific characteristics. More specifically, our PF-based video decoders utilize the size of each frame for the prediction of its decoding time. Compared to previous work, the PF is more suitable for our purpose, since it achieves higher prediction accuracy, even for highly nonstationary workloads such as H.264 clips. Our results show that the energy saved by the proposed approach is comparable to that of the ideal policy called oracle-DVFS, while the existing methods we tested were far inferior to oracle-DVFS in terms of H.264 video decoding. Additionally, when our method was used, only 0.40 and 6.88 percent of the frames missed their deadlines with negligible computational overhead for MPEG and H.264, respectively. [ABSTRACT FROM AUTHOR]

Viperin is an important virus-induced protein in animals that negatively participates in RNA viral replication and transcription. The reactive machinery of viperin suggests that it produces a regulatory molecule ddhCTP, which may affect immune regulation. In this study, we investigated the expression pattern of viperin in larval and adult stages of zebrafish by whole-mount in situ hybridization and reverse transcription-quantitative PCR (RT-qPCR). To elucidate the function of viperin, we generated a zebrafish knockout model using the CRISPR/Cas9 method and evaluated the mutation's effects under viral hemorrhagic septicemia virus (VHSV) infections. In zebrafish larvae, viperin was expressed in the brain region, eye, and pharynx, which was confirmed by cryosectioning. In adult zebrafish, blood cells showed the highest levels of viperin expression. In 5 dpf fish challenged with VHSV, the expression of the viral NP protein was significantly enhanced in viperin−/− compared to wild-type fish. In vitro VHSV propagation analysis indicated comparatively higher levels of virus propagation in viperin−/− fish. Mortality analysis confirmed higher mortality rates, and interferon gene expression analysis showed a strong upregulation of interferon (ifn)φ1 and 3 gene in viperin−/− fish infected with VHSV. This study describes the successful generation of a viperin-knockout model and the role of viperin during VHSV infections. [Display omitted] • A zebrafish knockout model (KO) for the viperin gene was constructed by CRISPR/Cas9-mediated genome editing. • Zebrafish viperin expression was analyzed by in situ hybridization and cryosectioning. • Fish morphology, virus tropism and titer were analyzed in VHSV-infected KO fish. • Interferon system and neutrophile migration were studied. [ABSTRACT FROM AUTHOR]

We describe a simple hydrothermal process for synthesizing vaterite-type CaCO 3 microparticles with a hexagonal bipyramidal shape using seawater. Vaterite with a spherical morphology is synthesized at room temperature relatively frequently, and a few high-temperature synthetic methods have been reported. However, the majority of these procedures require specific conditions, such as the use of detrimental surfactants, ultrasonication, and microwave irradiation. In this study, we successfully synthesized vaterite in Mg-free seawater using Ca(NO 3) 2 and urea, which do not require surfactants/organic additives or complex experimental conditions. The obtained vaterite hexagonal bipyramids had sharp edges and facets and a purity of 96%. We investigated the effects of the solvent type, reaction temperature, reaction time, and the ratio of Ca2+ and CO 3 2−, on the crystallization and morphology of the CaCO 3 polymorph. The phase and morphology of the vaterite were primarily determined by the synergistic effects of seawater and the ammonium ions from urea. [Display omitted] • Vaterite CaCO 3 was synthesized by Mg-free seawater aided hydrothermal method. • Unique hexagonal bipyramidal morphology was observed for the vaterite particles. • No organic additive/solvent is used for hexagonal bipyramidal vaterite formation. • Temperature (T) and time (t) play major roles in the vaterite formation. • CaCO 3 polymorphs follow same phase transition route irrespective of T and t. [ABSTRACT FROM AUTHOR]

This study was performed to understand how chlorpyrifos (CHL) affects zebrafish ( Danio rerio ) embryos and adults, by exposing this model organism to various concentrations of the insecticide. The 96-h acute toxicity test to determine the effect of CHL on adult zebrafish yielded a LC 50 of 709.43 μg/L −1 . Small molecular weight proteins less than 25 kDa and phospholipids were analyzed with MALDI-TOF MS/MS in order to compare expression patterns, revealing that some peaks were dramatically altered after CHL treatment. Whereas no acute toxicity was detected in the embryo toxicity test, malformation of zebrafish larvae was observed, with many individuals harboring curved spines. In an angiogenesis test on larvae of transgenic zebrafish, CHL did not have an inhibitory effect. Relative gene expression analyses using real-time polymerase chain reaction (RT-PCR) of DNA from zebrafish embryos revealed that different subtypes of cytochrome P450 (CYP450), such as CYP1A and CYP3A, were significantly up-regulated in response to CHL at a concentration of 400 μg/L −1 compared to the control. The expression level of NR1I2 , a CYP gene transcriptional regulator, UGT1a1, and MDR1 were all up-regulated in the CHL-treated embryos. Finally, the expression level of acetylcholinesterase (AChE) and catalase (CAT) decreased, whereas that of superoxide dismutase (SOD) did not differ significantly. Our results suggest that the up-regulation of metabolic enzymes including CYP450 and MDR1 may be involved in CHL resistance in zebrafish. [ABSTRACT FROM AUTHOR]

Herein, we report that the tungsten trioxide (WO 3) nanosheets and 2%Ag/15%WO 3 /HNT (Ag/WO 3 /HNT) composite photocatalysts were synthesized by the hydrothermal process and solvent evaporation methods. Various characterization techniques such as XRD, FT-IR, FE-SEM, EDS, HR-TEM, and UV-DRS were used to investigate the bulk and surface properties of bare and composite photocatalysts. The photocatalytic performance of halloysite nanotube (HNT), WO 3 /HNT, and Ag/WO 3 /HNT composite photocatalysts were evaluated by methylene blue (MB) dye degradation efficiency under solar light irradiation for 90 min. Among the various catalysts, Ag/WO 3 /HNT photocatalyst showed significant photocatalytic performance (96% degradation for MB dye). The existence of Ag indicates that surface plasmon resonance (SPR) impacts the developed composite and acts as an electron mediator. The scavenger experiment showed that the superoxide radical anions play a significant role in MB degradation under solar light with the most active Ag/WO 3 /HNT photocatalyst. The degradation of the MB dye has been monitored by Total Organic Carbon (TOC) measurements. The degradation intermediates of MB were analyzed by GC-MS, and a suitable degradation pathway was proposed. The most active Ag/WO 3 /HNT photocatalyst was also found to be the most stable and recyclable. The trapping experiment with Ag/WO 3 /HNT composite catalyst has led to the development of a dye degradation mechanism. [Display omitted] • Bare WO 3 and Ag/WO 3 /HNT composite catalyst were synthesized. • Ag/WO 3 /HNT photocatalyst showed significant photocatalytic performance with 96% degradation for MB. • Excellent stability and recyclability of Ag/WO 3 /HNT photocatalyst were observed. • A possible photocatalytic mechanism of the most active Ag/WO 3 /HNT catalyst was proposed. [ABSTRACT FROM AUTHOR]

Tumor necrosis factor (TNF) is an important cytokine involved in immune responses to bacterial infections in vertebrates, including fish. Although Tnf-α is a well-studied cytokine, there are contradictory findings about Tnf-α function following bacterial infection. In this study, we analyzed the expression and function of the Tnf-α-type I isoform (Tnf-α1) in zebrafish by knockout experiments using the CRISPR/Cas9 gene-editing tool. The open reading frame of tnf-α1 encodes a 25.82 kDa protein with 234 amino acids (aa). The expression of tnf-α1 in the early stages of zebrafish was observed from the 2-cell stage. Adult zebrafish spleens showed the highest expression of tnf-α1. To evaluate the function of Tnf-α1, an 8 bp deletion in the target region, resulting in a short truncated protein of 55 aa, was used to create the tnf-α1 knockout mutant. The pattern of downstream gene expression in 7-day larvae in wild-type (WT) and tnf-α1 knockout fish was examined. We also verified the fish mortality rate after Edwardsiella piscicida challenge and found that it was much higher in tnf-α1 knockout fish than in WT fish. Additionally, downstream gene expression analyses after E. piscicida exposure revealed a distinct expression pattern in tnf-α1 knockout fish compared to that in WT fish. Overall, our study using tnf-α1 deletion in zebrafish confirmed that Tnf-α1 is critical for immune regulation during bacterial infection. • The tnf-α1 expression pattern was analyzed in different developmental stages and tissues in zebrafish. • The knockout tnf-α1 zebrafish was generated by CRISPR/Cas9-mediating genome editing tool. • The tnf-α1 KO had impaired immunity to E. piscicida infection, as evidenced by their mortality and study of downstream genes. [ABSTRACT FROM AUTHOR]

Objective: Food insecurity in college students is associated with poor eating habits. The purpose of this project was to increase consumption of fruits and vegetables in college students using a campus food pantry. Participants: Twenty-nine college students completed the study. Methods: Participants viewed weekly food preparation videos demonstrating cost-effective recipes containing fruits and vegetables and a key take-away message. Participants were provided a recipe and food to cook at home based on the Cooking Matters Food Pantry Toolkit. A 24-h dietary recall was collected pre- and postintervention using the Automated Self-Administered 24-H Dietary Assessment. This descriptive comparative study used repeated measure ANOVA. Results: Females (n = 19) increased their fruit and vegetable consumption by 1.3 cups daily (p <.05) while males (n = 10) decreased fruit and vegetable consumption at the postintervention measurement. Conclusion: Increased consumption of fruits and vegetables in female college students using the food pantry was a modifiable behavior in this study. [ABSTRACT FROM AUTHOR]

Paired Box (Pax) gene family, a group of transcription regulators have been implicated in diverse physiological processes. However, their role during hematopoiesis which generate a plethora of blood cells remains largely unknown. Using a previously reported single cell transcriptomics data, we analyzed the expression of individual Pax family members in hematopoietic cells in zebrafish. We have identified that Pax9, which is an essential regulator for odontogenesis and palatogenesis, is selectively localized within a single cluster of the hematopoietic lineage. To further analyze the function of Pax9 in hematopoiesis, we generated two independent pax9 knock-out mutants using the CRISPR-Cas9 technique. We found that Pax9 appears to be an essential regulator for granulopoiesis but dispensable for erythropoiesis during development, as lack of pax9 selectively decreased the number of neutrophils with a concomitant decrease in the expression level of neutrophil markers. In addition, embryos, where pax9 was functionally disrupted by injecting morpholinos, failed to increase the number of neutrophils in response to pathogenic bacteria, suggesting that Pax9 is not only essential for developmental granulopoiesis but also emergency granulopoiesis. Due to the inability to initiate emergency granulopoiesis, innate immune responses were severely compromised in pax9 morpholino-mediated embryos, increasing their susceptibility and mortality. Taken together, our data indicate that Pax9 is essential for granulopoiesis and promotes innate immunity in zebrafish larvae. • We identified Pax9 as a critical regulator for hematopoiesis in zebrafish. • Pax9 is essential for granulopoiesis but dispensable for erythropoiesis in zebafish. • Pax9 is a key modulator for innate immune responses in zebrafish. [ABSTRACT FROM AUTHOR]

[Display omitted] • Ca precursor anions play an important role in the morphology of CaCO 3 superstructures. • The synergistic effects of anions, seawater, and ammonium ions alter the morphology of CaCO 3. • No additives/organic solvent is used for metastable CaCO 3 phase formation. • Selective crystallization of CaCO 3 polymorph was done by seawater-aided hydrothermal method. This work provides crucial insights into the crystallization and morphogenesis of CaCO 3 polymorphs in seawater at elevated temperatures. Using Ca source anions as a controlling factor, we modify the crystallization of CaCO 3 and demonstrate that stable and intricate particle morphologies of CaCO 3 , such as hexagonal bipyramidal, disks, and twin structures, can be generated in seawater without surfactants. The synthesis of CaCO 3 was accomplished through a direct hydrothermal method utilizing water-soluble Ca salts (calcium chloride, calcium nitrate, and calcium acetate) as Ca source and urea as a carbonate source. Depending on the specific source anions used, various morphological changes were observed. The CaCO 3 prepared with the nitrate source yields vaterite as the major polymorph with hexagonal bipyramidal morphology. Pseudohexagonal prisms of aragonite were the primary product obtained with calcium acetate as the source. In contrast, chloride ions yielded vaterite and aragonite depending on the reaction temperature and time. The purity of CaCO 3 prepared with seawater was > 94 % regardless of the Ca source. We observe that the phase formation follows the order of gypsum-vaterite-aragonite with an increase in hydrothermal reaction time. Further, we propose the formation mechanism of vaterite hexagonal bipyramidal and aragonite pseudohexagonal prism superstructures where directional aggregation of particles occurs in the presence of ammonium ions and seawater solvent. These findings, obtained through the exploration of seawater and different Ca sources, offer a comprehensive understanding of how source anions influence the selective formation of CaCO 3 polymorph and its morphological tuning. This information can be applied to tailor the synthesis of CaCO 3 for specific applications. [ABSTRACT FROM AUTHOR]

Myeloid differentiation primary response protein-88 (MYD88) is an essential adaptor molecule in pathogen-related pattern recognition signaling pathways. Toll-like and interleukin receptors recognize numerous signals and are funneled through MyD88 to express genes responsible for the innate and adaptive immune systems. In the present study, the relevance of MyD88 in viral hemorrhagic septicemia virus (VHSV) was investigated by generating myd88 −/− zebrafish. The model was challenged with VHSV, and viral propagation was quantified by evaluating clinical symptoms, mortality, and VHSV copy number. The infected fish showed abnormal morphologies, such as subcutaneous hemorrhages, abdominal swelling, and bulging eyes, which were comparatively more intense in myd88 −/− fish than in the wild-type. An injury infection experiment conducted in zebrafish larvae indicated a substantial spread of VHSV in the wound site. The number of neutrophils and macrophages recruited to the wounded area were markedly reduced in myd88 −/− fish. According to gene expression analysis, VHSV NP gene expression was considerably upregulated in myd88 −/− fish. Substantial gene expression and immune cell marker modulation were observed in the mutant model compared to that in the wild-type. These results suggest that the lack of a significant adaptor protein for immune signal transduction results in enhanced VHSV replication. • A myd88 ablated zebrafish with a 4 bp frameshift mutation was generated by CRISPR/Cas9 genomic engineering. • myd88 KO fish had lower survival and higher VHSV titers during the VHSV infection experiment. • A lack of myd88 impairs the rapid immune responses to VHSV. • The tail injury model of myd88 ablated fish had higher virus penetration. • myd88 ablation significantly impaired the immune cell dynamics. [ABSTRACT FROM AUTHOR]

In the present work, it is noticed that the synthesis of photocatalytic bare and composite materials: i) bare graphitic carbonitride g-C 3 N 4 (GCN), ii) modified with tungsten oxide WO 3 /g-C 3 N 4 (WGCN), and iii) with both tungsten oxide and silver nanoparticles WO 3 /Ag/g-C 3 N 4 (WAgGCN), by three different methods: calcination, precipitation, and ultrasonication. Also, their photocatalytic efficiencies were evaluated towards the photodegradation of different dyes under visible, UV, and sunlight irradiations with promising results. Here, the WAgGCN composite showed significant photocatalytic performance to decolorized Methylene Blue (98%) and Crystal Violet (95%), together with high chemical stability and excellent recyclability. The presence of Ag shows the influences of surface plasmon resonance and acts as an electron mediator in the formed heterostructures. The photogenerated holes performed vital roles in the catalytic degradation of dye solutions over the WAgGCN composite catalyst, confirmed by radical trapping experiments. Total Organic Carbon (TOC) and Chemical Oxygen Demand (COD) measurements confirmed the mineralization of the dyes. • Bare GCN, WGCN, and WAgGCN heterojunction photocatalysts were synthesized. • The WAgGCN composite photocatalyst demonstrated excellent photocatalytic efficiency. • Enhanced degradation and mineralization of MB and CV dyes were achieved. • Excellent stability and recyclability of WAgGCN photocatalyst were observed. [ABSTRACT FROM AUTHOR]

Our previous report revealed that Gardenia jasminoides (GJ) has protective effects against acute pancreatitis. So, we examined whether aqueous extract of GJ has anti-inflammation and antifibrotic effects even against cerulein-induced chronic pancreatitis (CP). CP was induced in mice by an intraperitoneal injection of a stable cholecystokinin (CCK) analogue, cerulein, six times a day, four days per week for three weeks. GJ extract (0.1 or 1 g/kg) or saline (control group) were intraperitoneally injected 1 h before first cerulein injection. After three weeks of stimulation, the pancreas was harvested for the examination of several fibrotic parameters. In addition, pancreatic stellate cells (PSCs) were isolated using gradient methods to examine the antifibrogenic effects of GJ. In the cerulein-induced CP mice, the histological features of the pancreas showed severe tissue damage such as enlarged interstitial spaces, inflammatory cell infiltrate and glandular atrophy, and tissue fibrosis. However, treatment of GJ reduced the severity of CP such as pancreatic edema and inflammatory cell infiltration. Furthermore, treatment of GJ increased pancreatic acinar cell survival, and reduced pancreatic fibrosis and activation of PSC in vivo and in vitro. In addition, GJ treatment inhibited the activation of c-Jun N-terminal kinase (JNK) and extracellular signal-regulated protein kinase (ERK) in the PSCs. These results suggest that GJ attenuated the severity of CP and the pancreatic fibrosis by inhibiting JNK and ERK activation during CP. [ABSTRACT FROM AUTHOR]

The interferon-induced GTP-binding protein Mx is responsible for a specific antiviral state against a broad spectrum of viral infections that are induced by type-I interferons (IFN α/β) in different vertebrates. In this study, the Mx gene was isolated from the constructed mullet cDNA database. Structural features of mullet Mx (MuMx) were analyzed using different in-silico tools. The pairwise comparison revealed that the MuMx sequence was related to Stegastes partitus Mx with an 83.7% sequence identity, whereas MuMx was clustered into the teleost category in the phylogentic analysis. Sequence alignment showed that the dynamin-type guanine nucleotide-binding domain (G_DYNAMIN_2), central interactive domain (CID), and GTPase effector domain (GED) were conserved among Mx counterparts. The transcriptional expression of MuMx was the highest in blood cells from unchallenged fish. The temporal mRNA profile showed that MuMx expression was significantly elevated in all tissues, including blood, spleen, head kidney, liver, and gills after the injection of polyinosinic-polycytidylic acid (poly I:C) at many time points. Moreover, MuMx expression increased slightly, in the blood, spleen, and head kidney at a few time points after the injection of lipopolysaccharide (LPS) and Lactococcus garvieae (L. garvieae). Results of the subcellular localization analysis confirmed that the MuMx protein was highly expressed in the cytoplasm. The analysis of the gene expression of the viral hemorrhagic septicemia virus (VHSV) under conditions of MuMx overexpression confirmed the significant inhibition of viral transcripts. The cell viability (MTT) assay and VHSV titer quantification with the presence of MuMx indicated a significant reduction in virus replication. Collectively, these findings suggest that Mx is a specific immune-related gene that elicits crucial antiviral functions against viral antigens in the mullet fish. • Mx was identified from the constructed red lip mullet cDNA data base. • Antiviral activity of MuMx was investigated by antiviral activity assay. • Cell survival rate was evaluated by MTT assay in the presence of VHSV. • Transcriptional level of MuMx was modulated by pathogenic stimulants. • The subcellular localization of MuMx was observed in the cytoplasm. [ABSTRACT FROM AUTHOR]

The major role of inner medullary collecting duct (IMCD) cells is to maintain water and sodium homeostasis. In addition to the major role, it also participates in the protection of renal and systemic inflammation. Although IMCD cells could take part in renal and systemic inflammation, investigations on renal inflammation in IMCD cells have rarely been reported. Although berberine (BBR) has been reported to show diverse pharmacological effects, its anti-inflammatory and protective effects on IMCD cells have not been studied. Therefore, in the present study, we examined the anti-inflammatory and protective effects of BBR in mouse IMCD-3 (mIMCD-3) cells against lipopolysaccharide (LPS). An MTT assay was carried out to investigate the toxicity of BBR on mIMCD-3 cells. Reverse transcription quantitative-PCR and western blotting were performed to analysis pro-inflammatory molecules and cytokines. Mechanisms of BBR were examined by western blotting and immunocytochemistry. According to previous studies, pro-inflammatory molecules, such as inducible nitric oxide synthase and cyclooxygenase-2, and pro-inflammatory cytokines, such as interleukin (IL)-1β, IL-6 and tumor necrosis factor-α are increased in LPS-exposed mIMCD-3 cells. However, the production of these pro-inflammatory molecules is significantly inhibited by treatment with BBR. In addition, BBR inhibited translocation of nuclear factor (NF)-κB p65 from the cytosol to the nucleus, and degradation of inhibitory κ-Bα in LPS-exposed mIMCD-3 cells. In conclusion, BBR could inhibit renal inflammatory responses via inhibition of NF-κB signaling and ultimately contribute to amelioration of renal injury during systemic inflammation. [ABSTRACT FROM AUTHOR]

The transcription factor, activator protein-1 (AP-1), is a dimeric protein and a downstream member of the mitogen-activated protein kinase (MAPK) signaling pathway. It regulates a wide array of functions including, cell proliferation, survival, differentiation, response to UV-irradiation, immune responses, and inflammatory conditions. AP-1 belongs to the basic leucine zipper (bZIP) protein family, which consists of members from Jun, Fos, Maf, and ATF subfamilies. In the present study, c-Jun and c-Fos homologs were identified from a transcriptome database of Liza haematocheila and designated as Lhc-Jun and Lhc-Fos. In both sequences, the signature bZIP domain was identified and also the DNA binding sites, dimerization sites, as well as the phosphorylation sites, were found to be highly conserved through evolution. Tissue distribution analysis revealed that both Lhc-Jun and Lhc-Fos transcripts were ubiquitously expressed in all examined tissues of healthy mullets. In order to determine the transcriptional modulations of Lhc-Jun and Lhc-Fos, challenge experiments were carried out using LPS, poly I:C, and L. garvieae. The qRT-PCR analysis revealed significant upregulation of Lhc-Jun and Lhc-Fos in blood, gill, liver, and spleen. This is the first study that explores the correlation between UV-irradiation and AP-1 ortholog expression in teleosts. Also, this is the first time that the functional characterization of the teleost c-Fos ortholog has been carried out. Sub-cellular localization of Lhc-Jun and Lhc-Fos was observed in the nucleus. AP-1-Luc reporter assays revealed significant higher luciferase activities in both Lhc-Jun and Lhc-Fos proteins compared to mock controls. These results strongly suggest that Lhc-Jun and Lhc-Fos might play a significant role in Liza haematocheila immunity by regulating AP-1 promoter sequences in immune and stress-related genes. • Homologs of c-Jun and c-Fos were identified from Liza haematocheila (Lhc-Jun and Lhc-Fos). • Lhc-Jun and Lhc-Fos shared signature bZIP domain architecture. • Both genes were highly induced upon PAMPs and live pathogen challenges. • UV irradiation results a unique expression pattern of both genes in MK cells. • Both recombinant proteins were nuclear localized and increased the AP-1 activity. [ABSTRACT FROM AUTHOR]

Abstract Heme oxygenase-1 (HO-1) has an anti-inflammatory action in acute pancreatitis (AP). However, its mechanism of action and natural compounds/drugs to induce HO-1 in pancreas are not well understood. In this study, we investigated the regulatory mechanisms of HO-1 during AP using desoxo-narchinol-A (DN), the natural compound inducing HO-1 in the pancreas. Female C57/BL6 Mice were intraperitoneally injected with supramaximal concentrations of cerulein (50 μg/kg) hourly for 6 h to induce AP. DMSO or DN was administered intraperitoneally, then mice were sacrificed 6 h after the final cerulein injection. Administration of DN increased pancreatic HO-1 expression through activation of activating protein-1, mediated by mitogen-activated protein kinases. Furthermore, DN treatment reduced the pancreatic weight-to-body weight ratio as well as production of digestive enzymes and pro-inflammatory cytokines. Inhibition of HO-1 by tin protoporphyrin IX abolished the protective effects of DN on pancreatic damage. Additionally, DN treatment inhibited neutrophil infiltration into the pancreas via regulation of chemokine (C-X-C motif) ligand 2 (CXCL2) by HO-1. Our results suggest that DN is an effective inducer of HO-1 in the pancreas, and that HO-1 regulates neutrophil infiltration in AP via CXCL2 inhibition. Graphical abstract Unlabelled Image Highlights • Desoxo-narchinol-A (DN) is a natural compound of HO-1 inducer in pancreas. • Mechanism of DN-induced HO-1 is mediated by MAPK/Activator Protein-1/HO-1 signaling. • DN-induced HO-1 blocks neutrophil infiltration into pancreas via inhibition of CXCL2. • DN inhibits cerulein-induced acute pancreatitis (AP) and AP-associated lung injury. [ABSTRACT FROM AUTHOR]

Abstract Interleukin-10 (IL-10) is a pleiotropic cytokine involved in the regulation of innate and adaptive immunity. In this study, IL-10 from big-belly seahorse (Hippocampus abdominalis) (HaIL-10) was characterized based on its molecular and functional aspects. The coding sequence of HaIL-10 is 570 bp in length and encodes a 189-amino acid residue protein (calculated molecular weight, 21.89 kDa). The deduced amino acid sequence comprises a typical signal peptide and a mature peptide domain sequence carrying four conserved Cys residues and two additional Cys residues specific to fish. Phylogenetic analysis indicated an evolutionary relationship between HaIL-10 and its counterparts in other vertebrates, with close clustering to the fish-specific homologs. Recombinant HaIL-10 (rHaIL-10) significantly reduced nitric oxide (NO) production by lipopolysaccharide (LPS)-induced murine macrophage RAW 264.7 cells in a concentration-dependent manner but had no effect on cell viability, suggestive of its involvement in immune response. The protein expressions of iNOS and COX-2 were significantly reduced by rHaIL-10 in LPS-induced murine macrophages RAW 264.7 cells. HaIL-10 mRNA expression was observed in all analyzed tissues, with the maximum expression being noted in the kidney and ovary. However, transcriptional levels of HaIL-10 were significantly higher in the blood, gill, and intestine upon in vivo induction with LPS, polyinosinic:polycytidylic acid [poly (I:C)], and Streptococcus iniae. To summarize, our findings help in the improved understanding of the biological functions of HaIL-10 and modulation of HaIL-10 mRNA expression in response to immune stress. Highlights • Identified and characterized interleukin-10 from big-belly seahorse • The HaIL-10 was constitutively expressed in healthy seahorse tissues. • mRNA expression pattern found to be correlated with the immune stresses. • rHaIL-10 reduces NO production in LPS-induced macrophage RAW 264.7 cells. • rHaIL-10 reduces iNOS and COX-2 proteins in LPS-induced macrophage RAW 264.7 cells. [ABSTRACT FROM AUTHOR]

Abstract Complement system orchestrates the innate and adaptive immunity via the activation, recruitment, and regulation of immune molecules to destroy pathogens. However, regulation of the complement is essential to avoid injuries to the autologous tissues. The present study unveils the characteristic features of an important complement component, anaphylatoxin inactivator from red lip mullet at its molecular and functional level. Mullet carboxypeptidase N1 (MuCPN1) cDNA sequence possessed an open reading frame of 1347 bp, which encoded a protein of 449 amino acids with a predicted molecular weight of 51 kDa. In silico analysis discovered two domains of PM14-Zn carboxypeptidase and a C-terminal domain of M14 N/E carboxypeptidase, two zinc-binding signature motifs, and an N-glycosylation site in the MuCPN1 sequence. Homology analysis revealed that most of the residues in the sequence are conserved among the other selected homologs. Phylogeny analysis showed that MuCPN1 closely cladded with the Maylandia zebra CPN1 and clustered together with the teleostean counterparts. A challenge experiment showed modulated expression of MuCPN1 upon polyinosinic:polycytidylic acid and Lactococcus garviae in head kidney, spleen, gill, and liver tissues. The highest upregulation of MuCPN1 was observed 24 h post infection against poly I:C in each tissue. Moreover, the highest relative expressions upon L. garviae challenge were observed at 24 h post infection in head kidney tissue and 48 h post infection in spleen, gill, and liver tissues. MuCPN1 transfected cells triggered a 2.2-fold increase of nitric oxide (NO) production upon LPS stimulation compared to the un-transfected controls suggesting that MuCPN1 is an active protease which releases arginine from complement C3a, C4a, and C5a. These results have driven certain way towards enhancing the understanding of immune role of MuCPN1 in the complement defense mechanism of red lip mullet. Highlights • Carboxypeptidase N1 complement component was identified from the red lip mullet. • Ubiquitous expression of MuCPN1 was observed in healthy mullet tissues. • Modulated transcriptions of MuCPN1 revealed the importance in the immune responses. • MuCPN1 was enhanced the nitric oxide production at an inflammatory condition. [ABSTRACT FROM AUTHOR]

A batch of wild common carp and largemouth bass died in Andong, Gyeongsangbuk-do province, South Korea, in 2016. Moribund fish showed typical signs of spring viremia of carp (SVC) disease, which causes acute hemorrhage in the skin and ascites. Thus far, SVC disease has been detected in several regions of the world but never in South Korea. Suspecting the infectious agent to be the SCV virus (SVCV), the moribund fish were sampled and screened. The isolated virus developed a cytopathic effect in EPC cells. Both viral isolates from the common carp (ADC-SVC2016-1) and largemouth bass (ADC-SVC2016-3) were identical in terms of their genome sequence, which were 11,034 bp nucleotides in length. Genome comparison exhibited greater sequence similarity with the Asian SVCV sequences available at NCBI. Phylogenetic analysis revealed that the Korean SVCV isolates were clustered within the Asian clade. More specifically, evolutionary analysis by using the P gene sequences showed that the Korean isolates were sub-cladded within the Iai genogroup but diverged from Chinese strains of SH150514 and SH160901. The Korean isolates shared more than 98% sequence similarity with the two Chinese SVCV isolates, suggesting that the spread of SVCV originated from China. The isolated virus had cytopathic effects on EPC cells. Virus transmission studies showed that the virus exhibited the highest virulence at 15 °C, which was also dependent on the method used, with the injection method being better than the immersion and cohabitation methods. This is the first study to document that Korean SVCV isolates may be epizootic in wild common carp and other susceptible animal populations in South Korea. [ABSTRACT FROM AUTHOR]

C1-inhibitor (C1inh) plays a crucial role in assuring homeostasis and is the central regulator of the complement activation involved in immunity and inflammation. A C1-inhibitor gene from Sebastes schlegelii was identified and designated as SsC1inh . The identified genomic DNA and cDNA sequences were 6837 bp and 2161 bp, respectively. The genomic DNA possessed 11 exons, interrupted by 10 introns. The amino acid sequence possessed two immunoglobulin-like domains and a serpin domain. Multiple sequence alignment revealed that the serpin domain of SsC1inh was highly conserved among analyzed species where the two immunoglobulin-like domains showed divergence. The distinctiveness of teleost C1inh from other homologs was indicated by the phylogenetic analysis, genomic DNA organization, and their extended N-terminal amino acid sequences. Under normal physiological conditions, SsC1inh mRNA was most expressed in the liver, followed by the gills. The involvement of SsC1inh in homeostasis was demonstrated by modulated transcription profiles in the liver and spleen upon pathogenic stress by different immune stimulants. The protease inhibitory potential of recombinant SsC1inh (rSsC1inh) and the potentiation effect of heparin on rSsC1inh was demonstrated against C1esterase and thrombin. For the first time, the anti-protease activity of the teleost C1inh against its natural substrates C1r and C1s was proved in this study. The protease assay conducted with recombinant black rockfish C1r and C1s proteins in the presence or absence of rSsC1inh showed that the activities of both proteases were significantly diminished by rSsC1inh. Taken together, results from the present study indicate that SsC1inh actively plays a significant role in maintaining homeostasis in the immune system of black rock fish. [ABSTRACT FROM AUTHOR]

C1r and C1s are serine proteases responsible for activating the classical complement pathway to initiate the complement cascade, which plays a crucial role in eliminating invading pathogenic microbes. In this study, cDNA sequences of C1r and C1s were identified from black rockfish and designated as SsC1r and SsC1s , respectively. In both sequences, two CUB domains, an EGF-like domain, two CCP domains, and a trypsin-like serine protease domain were identified. Multiple sequence alignments with known vertebrate homologs demonstrated that both sequences were highly conserved and, especially, the catalytic and substrate binding residues were completely conserved. In the constructed phylogenetic tree, C1r and C1s formed two separate clusters, which further branched into groups of related organisms. SsC1r and SsC1s joined with their respective teleostean clusters. Transcriptional analysis showed that the highest mRNA expression level was in the liver under normal physiological conditions. Significantly upregulated expression of both mRNAs in spleen and liver after pathologic stress, by intraperitoneal injection with different stimuli, suggested their vital role in immunity. The serine protease domains of SsC1r and SsC1s were cloned and the recombinant proteins were expressed and purified. A protease assay, conducted to confirm their functionality, indicated that both recombinant proteins had proteolytic activity. Taken together, these results indicate that SsC1r and SsC1s have significant properties to aid in the immunity of black rockfish by activating the complement system by proteolytic cleavage. [ABSTRACT FROM AUTHOR]

Akirins, members of the NF-κB signaling pathway, are highly conserved nuclear proteins, which regulate gene expression in many physiological processes, including immunity, myogenesis, carcinogenesis, and embryogenesis. The akirin family in teleost fish consists of two to three genes. In the present study, three akirin genes from Hippocampus abdominalis were identified from a transcriptome database and designated as HaAkirin1 , HaAkirin2(1) , and HaAkirin2(2) . The nuclear localization of HaAkirin1 and HaAkirin2(1) was confirmed by subcellular localization analysis. In contrast, diffused localization of HaAkirin2(2) was identified in the nucleus and cytoplasm that confirmed the aberrant nature of the nuclear localization signal. Phylogenetic analysis revealed a closer relationship of HaAkirins with other known teleost akirins. All three HaAkirin transcripts were ubiquitously expressed in all examined tissues with higher expression in ovary tissue. Immune challenge with LPS, poly I:C, and Streptococcus iniae exhibited a significant increase in the expression of all three HaAkirins in kidney and liver tissues. NF-κB luciferase assays revealed that relative luciferase activity was significantly higher for all three HaAkirin genes than mock controls. These results suggest that HaAkirin genes might play a role in regulating NF-κB dependent immune gene expression and their expression could be induced by bacterial and viral pathogen recognition molecular patterns. [ABSTRACT FROM AUTHOR]

Viral hemorrhagic septicemia is a serious disease that can assume epidemic proportions in cultivated olive flounders in Korea. The causal agent of this disease is viral hemorrhagic septicemia virus (VHSV), and it leads to vast economic losses in the olive flounder aquaculture industry. Therefore, the rapid and accurate detection of VHSV is paramount. At present, the manual of the World Organization for Animal Health's Office International des Epizooties (OIE) for pathogen detection is being followed for VHSV detection in Korea. However, in that manual, the primers for VHSV detection are based on the European VHSV genotype Ia. In this study, we identified 5 VHSV strains from olive flounders in Korea, and the nucleotide and amino acid sequences of VHSV genes (N, P, M, and G) were compared to investigate the genetic variation of VHSV genotypes. As expected, VHSV isolates from Korea were highly related to genotype IVa, clearly differing from the 3 European genotypes. In addition, the N gene showed low genetic variation and therefore might be considered a useful marker for VHSV detection, regardless of genotype. In contrast, the other VHSV genes (P, M, and G) had different nucleotides within genotypes. They might be suitable for designing specific primers for distinguishing between various VHSV genotypes. Duplex RT-PCR using the newly designed primers successfully detected all VHSV isolates and validated the genotype (I and IV) without sequence analysis. According to the results, all the VHSV isolates were successfully detected, and their genotypes were validated by duplex RT-PCR using the newly designed primers. Collectively, our findings suggest that duplex PCR is a convenient and appropriate method for the diagnosis of VHSV isolates in Korean aquaculture systems. [ABSTRACT FROM AUTHOR]

Caveolins are principal membrane proteins of caveolae that play a central role in signal transduction, substrate transport, and membrane trafficking in various cell types. Numerous studies have reported the crucial role of caveolin-1 (CAV1) in response to invading microbes; yet, very little is known about molluscan CAV1. In this study, we identified and characterized CAV1 ortholog from the disk abalone, Haliotis discus discus (HdCAV1). The cDNA sequence of HdCAV1 is 826 bp long and encodes a 127-amino acid polypeptide. Characteristic caveolin superfamily domain (Glu 3 – Lys 126 ) and two possible transmembrane domains (Cys 48 - Tyr 67 and Ile 103 - Phe 120 ) were identified in the HdCAV1 protein. Homology analysis revealed that HdCAV1 shared higher identity (>47%) with molluscans, but lower identity with other species. Phylogenetic tree constructed by the neighbor-joining (NJ) method revealed a distinct evolutionary pathway for molluscans. Transcriptional analysis by SYBR Green qPCR showed the highest expression of HdCAV1 mRNA in late veliger stage, as compared to that in other embryonic developmental stages of disk abalone. In adult animals, gill tissue showed highest HdCAV1 transcript levels under normal physiological condition. Stimulations with two bacteria ( Vibrio parahaemolyticus and Listeria monocytogenes ), viral hemorrhagic septicemia virus, and two pathogen-associated molecular patterns (LPS and poly I:C) significantly modulated the expression of HdCAV1 transcripts. Collectively, these data suggest that CAV1 plays an important role in embryogenesis and host immune defense in disk abalone. [ABSTRACT FROM AUTHOR]

Cathepsin Z (CTSZ) is lysosomal cysteine protease of the papain superfamily. It participates in the host immune defense via phagocytosis, signal transduction, cell-cell communication, proliferation, and migration of immune cells such as monocytes, macrophages, and dendritic cells. Hence, CTSZ is also acknowledged as an acute-phase protein in host immunity. In this study, we sought to identify the CTSZ homolog from disk abalone (AbCTSZ) and characterize it at the molecular, genomic, and transcriptional levels. AbCTSZ encodes a protein with 318 amino acids and a molecular mass of 36 kDa. The structure of AbCTSZ reveals amino acid sequences that are characteristic of the signal sequence, pro-peptide, peptidase-C1 papain family cysteine protease domain, mini-loop, HIP motif, N -linked glycosylation sites, active sites, and conserved Cys residues. A pairwise comparison revealed that AbCTSZ shared the highest amino acid homology with its molluscan counterpart from Crassostrea gigas . A multiple alignment analysis revealed the conservation of functionally crucial elements of AbCTSZ, and a phylogenetic study further confirmed a proximal evolutionary relationship with its invertebrate counterparts. Further, an analysis of AbCTSZ genomic structure revealed seven exons separated by six introns, which differs from that of its vertebrate counterparts. Quantitative real time PCR (qPCR) detected the transcripts of AbCTSZ in early developmental stages and in eight different tissues. Higher levels of AbCTSZ transcripts were found in trochophore, gill, and hemocytes, highlighting its importance in the early development and immunity of disk abalone. In addition, we found that viable bacteria ( Vibrio parahaemolyticus and Listeria monocytogenes ) and bacterial lipopolysaccharides significantly modulated AbCTSZ transcription. Collectively, these lines of evidences suggest that AbCTSZ plays an indispensable role in the innate immunity of disk abalone. [ABSTRACT FROM AUTHOR]

Manganese superoxide dismutase (MnSOD) is a metaloenzyme that catalyzes dismutation of the hazardous superoxide radicals into less hazardous H 2 O 2 and H 2 O. Here, we identified a homolog of MnSOD from big belly seahorse ( Hippocampus abdominalis ; HaMnSOD ) and characterized its structural and functional features. HaMnSOD transcript possessed an open reading frame (ORF) of 672 bp which codes for a peptide of 223 amino acids. Pairwise alignment showed that HaMnSOD shared highest identity with rock bream MnSOD. Results of the phylogenetic analysis of HaMnSOD revealed a close proximity with rock bream MnSOD which was consistent with the result of homology alignment. The intense expression of HaMnSOD was observed in the ovary, followed by the heart and the brain. Further, immune related responses of HaMnSOD towards pathogenic stimulation were observed through bacterial and viral challenges. Highest HaMnSOD expression in response to stimulants Edwardsiella tarda , Streptococcus iniae , lipopolysaccharide (LPS), and polyinosinic-polycytidylic acid (Poly I:C) was observed in the late stage in the blood tissue. Xanthine/xanthine oxidase assay (XOD assay) indicated the ROS-scavenging ability of purified recombinant HaMnSOD (rHaMnSOD). The optimum conditions for the SOD activity of rHaMnSOD were pH 9 and the 25 °C. Collectively, the results obtained through the expressional analysis profiles and the functional assays provide insights into potential immune related and antioxidant roles of HaMnSOD in the big belly seahorse. [ABSTRACT FROM AUTHOR]

Chemokines are a family of chemotactic cytokines that regulate leukocyte migration. They are classified into four groups namely, CXC, CC, C and CX 3 C, based on the formation of a disulfide bridge. Among these, CXC chemokines have been identified as the largest group of chemokines in humans. In this study, we identified and functionally characterized a homolog of CXC chemokine from the big-belly seahorse, Hippocampus abdominalis , and designated it as ShCXCL. The cDNA of ShCXCL composed of a 342-bp open reading frame encoding 113 amino acids (aa). The CXC family-specific small cytokine domain (SCY) was identified from the mature peptide region, which comprised of a conserved CXC motif. As ShCXCL lacks an ELR (Glutamic acid-Leucine-Arginine) motif, it belongs to ELR − subfamily. The recombinant ShCXCL protein strongly induced the nitric oxide (NO) production in macrophage cells (RAW 264.7 cell line) and showed the chemotactic effect on flounder peripheral blood leukocytes. Tissue profiling showed a ubiquitous expression pattern in all examined tissues, with a high abundance in spleen. The up-regulated mRNA expression pattern of ShCXCL was observed in blood and kidney tissues after immune stimulation by live bacteria, such as Streptococcus iniae and Edwardsiella tarda , and mitogens, such as lipopolysaccharides (LPS) and polyinosinic:polycytidylic acid (poly I:C), suggesting its important role in host immune defense against microbial infection. [ABSTRACT FROM AUTHOR]

Apoptosis is a physiological process that can also participate in host immune defense mechanisms, including tumor growth suppression along with homeostasis and maturation of immune cells. Caspases are known to be involved in cellular apoptotic signaling; among them, caspase-8 plays an important role in the initiation phase of the apoptotic death cascade. In the current study, we molecularly characterized a caspase-8 homolog (designated as HaCasp-8) from Hippocampus abdominalis . The HaCasp-8 gene harbors a 1476 bp open reading frame (ORF) that codes for a protein of 492 amino acids (aa) with a predicted molecular mass of 55 kDa. HaCasp-8 houses the typical domain architecture of known initiator caspases, including the death effector domain and the carboxyl-terminal catalytic domain. As expected, phylogenetic analysis reflected a closer evolutionary relationship of HaCasp-8 with its teleostean similitudes. The results of our qPCR assays confirmed the ubiquitous expression of HaCasp-8 in physiologically important tissues examined, with pronounced expression levels in ovary tissues, followed by blood cells. HaCasp-8 expression at the mRNA level was found to be significantly modulated by lipopolysaccharide, polyinosinic:polycytidylic acid, Streptococcus iniae, and Edwardsiella tarda injection. Overexpression of HaCasp-8 could trigger a significant level of cell death in HEK293T cells, suggesting its putative role in cell death. Taken together, our findings suggest that HaCasp-8 is an important component in the caspase cascade, and its expression can be significantly modulated under pathogen stress conditions in the big-belly seahorse. [ABSTRACT FROM AUTHOR]

Aquaporin 8 (Aqp8) is a transmembrane protein that is selectively permeated by water and some small solutes, and physiologically contributes to acid-base equilibrium in the gastrointestinal tract. Here, we described the characterization and spatiotemporal expression pattern of zebrafish aqp8 ( zaqp8 ) gene family, including zaqp8a.1 , zaqp8a.2, and zaqp8b , during the early developmental stages. The expression of zaqp8a.1 started first in the lateral plate mesoderm at the 12-somite stage (ss) and then expanded sequentially to the dorsal aorta, intersegmental blood vessels and then to the dorsal longitudinal anastomotic vessel at 24 h post fertilization (hpf). At 28 hpf, expression of zaqp8a.1 was also detected in the embryonic heart tube. Four days post fertilization (dpf), strong zaqp8a.1 expression was detected in the gastrointestinal tract and liver. By 72 hpf, the expression of zaqp8a.2 was first detected in the primitive gut region but not detected in the liver. The expression of z a qp8b was first detected in the intermediate mesoderm at 10 ss. From 24 hpf to 6 dpf, the proximal convoluted segment of the embryonic kidney was marked by zaqp8b expression Overall, these differential expression patterns of aqp8a.1 , aqp8a.2, and aqp8b suggest that they possibly play distinct roles throughout the embryonic development by controlling or maintaining organ-specific cellular water homeostasis. Our study provides new evidence that organogenesis requires differential roles of Aqp8 proteins in zebrafish. [ABSTRACT FROM AUTHOR]