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Deubiquitinase cylindromatosis (CYLD) inhibits MAPK and NF-κB activation pathways by deubiquitinating upstream regulatory factors. Although CYLD has been identified and actively studied in mammals, nothing is known about its putative function in fish. In this study, we identified the gene encoding CYLD (OmCYLD) from rainbow trout, Oncorhynchus mykiss, and examined its role during pathogenic infections. The deduced amino acid sequence of OmCYLD contains conserved CAP-Gly and USP domains. In RTH-149 cells, the expression of OmCYLD was increased by stimulation with Edwardsiella tarda and Streptococcus iniae. Gain-of-function and loss-of-function experiments showed that OmCYLD down-regulates the activation of MAPK and NF-κB and the expression of pro-inflammatory cytokines in E. tarda-stimulated RTH-149 cells. These findings suggest that OmCYLD might function like those of mammals to negatively regulate bacteria-triggered signaling pathway in fish.

We report fabrication of low-density polymeric microparticles (micropolymofoam particles [μPFPs]) using the piezoelectric inkjet method. After ejecting a polymer solution prepared by dissolving polymers in a volatile organic solvent, microdroplets are periodically generated through an inkjet nozzle in the air. Rapid evaporation of the solvent as it falls through the air leads to a highly porous surface and internal morphology. Optical microscope observation confirms that the inside of the μPFPs is filled with air. Notably, the use of more than two different polymers results in kinetically stable particles, in which the heterogeneous polymer chains are randomly entangled. Shape transformation of the kinetically stable μPFPs occurs upon adding a swelling solvent, spontaneously changing their geometry from porous and rugged-shaped to a Janus-like structure with a clear phase boundary. This transformation is directly monitored using an optical microscope. The surface free energy calculation reveals that particles are transformed to adopt a thermodynamically stable structure; this is achieved by imparting fluidity to the randomly entangled polymer chains, which is energetically unfavorable. In addition, porosity control can be possible by selectively removing one polymer phase of kinetically stable particles comprising two different polymers. Furthermore, the developed inkjet method can be applied for producing composite particles encapsulated with functional nanoparticles and coating or patterning them directly onto a substrate.

Microbial bioremediation has gained attention as a cheap, efficient, and sustainable technology to manage the increasing environmental pollution. Since microorganisms in nature are not evolved to degrade pollutants, there is an increasing demand for developing safer and more efficient pollutant-scavengers for enhanced bioremediation. In this review, we introduce the strategies and technologies developed in the field of synthetic biology and their applications to the construction of microbial scavengers with improved efficiency of biodegradation while minimizing the impact of genetically engineered microbial scavengers on ecosystems. In addition, we discuss recent achievements in the biodegradation of fastidious pollutants, greenhouse gases, and microplastics using engineered microbial scavengers. Using synthetic microbial scavengers and multidisciplinary technologies, toxic pollutants could be more easily eliminated, and the environment could be more efficiently recovered.

Glutathione (GSH)-deprived Dictyostelium discoideum accumulates methylglyoxal (MG) and reactive oxygen species (ROS) during vegetative growth. However, the reciprocal effects of the production and regulation of these metabolites on differentiation and cell motility are unclear. Based on the inhibitory effects of γ-glutamylcysteine synthetase ( gcsA ) disruption and GSH reductase ( gsr ) overexpression on aggregation and culmination, respectively, we overexpressed GSH-related genes encoding superoxide dismutase (Sod2), catalase (CatA), and Gcs, in D. discoideum . Wild-type KAx3 and gcsA -overexpressing ( gcsA OE ) slugs maintained GSH levels at levels of approximately 2.1-fold less than the reference GSH synthetase-overexpressing mutant; their GSH levels did not correlate with slug migration ability. Through prolonged KAx3 migration by treatment with MG and H 2 O 2 , we found that MG increased after the mound stage in this strain, with a 2.6-fold increase compared to early developmental stages; in contrast, ROS were maintained at high levels throughout development. While the migration-defective sod2 - and catA -overexpressing mutant slugs ( sod2 OE and catA OE ) decreased ROS levels by 50% and 53%, respectively, these slugs showed moderately decreased MG levels (36.2 ± 5.8 and 40.7 ± 1.6 nmol g −1 cells wet weight, P 0.05) compared to the parental strain (54.2 ± 3.5 nmol g −1 ). Importantly, defects in the migration of gcsA OE slugs decreased MG considerably (13.8 ± 4.2 nmol g −1 , P 0.01) along with a slight decrease in ROS. In contrast to the increase observed in migrating sod2 OE and catA OE slugs by treatment with MG and H 2 O 2 , the migration of gcsA OE slugs appeared unaffected. This behavior was caused by MG-triggered Gsr and NADPH-linked aldolase reductase activity, suggesting that GSH biosynthesis in gcsA OE slugs is specifically used for MG-scavenging activity. This is the first report showing that MG upregulates slug migration via MG-scavenging-mediated differentiation.

The effects of diclofenac, a nonsteroidal anti-inflammatory drug (NSAID), on the fast tetrodotoxin-sensitive (TTX-S) and the slow tetrodotoxin-resistant (TTX-R) sodium currents in rat dorsal root ganglion neurons were investigated using the whole-cell patch-clamp method. Diclofenac suppressed both sodium currents in a dose-dependent manner. The apparent dissociation constants for the diclofenac suppression of TTX-S and TTX-R sodium currents were estimated to be 14 and 97 μM, respectively, at a holding potential of −80 mV. Diclofenac had no effect on the kinetic parameters of the activation process in either type of sodium current. However, diclofenac produced shifts of the steady-state inactivation curves in the hyperpolarizing direction in both types of sodium currents in a dose-dependent manner. At sufficiently negative holding potentials, the inhibitory effects of diclofenac on both types of sodium currents were minimal. The results suggested that diclofenac might bind to sodium channels with a greater affinity when they are in the inactivated state than when they are in the resting state. Effects of other NSAIDs (acetylsalicylic acid, antipyrin, indomethacin and flufenamic acid) on sodium currents were tested. Among these, only flufenamic acid suppressed the sodium currents to a considerable extent. Thus, the chemical structure of each NSAID, not the inhibition of cyclooxygenase, seems to be an important determinant in the sodium current inhibition. The suppression of sodium currents in sensory neurons by diclofenac and flufenamic acid would contribute to their analgesic activity in addition to their inhibition of cyclooxygenase.

The HP1043 protein of Helicobacter pylori is related to the two-component system that regulates prokaryotic transcription in response to environmental changes. The HP1043 gene encodes an orphan response regulator (RR). The N-terminal domain of HP1043 (HP1043N) was purified by affinity-column chromatography. The purified protein was crystallized by hanging drop vapor diffusion at 293 K. The crystal belongs to primitive orthorhombic space group P2 1 2 1 2 with cell dimensions of a = 89.0, b = 41.3, and c = 31.7 A. Assuming the presence of one molecule per asymmetric unit, the solvent content was estimated to be about 45%. A complete data set was collected at 1.8 A resolution.

The action of alprenolol, a non-selective beta blocker as well as 5-HT receptor antagonist, on the cloned cardiac human ether-a-go-go-related gene (HERG) channels was investigated using the whole-cell patch-clamp technique. Alprenolol reduced HERG whole-cell currents in a reversible concentration-dependent manner, with an IC50 value and a Hill coefficient of 12.3 ± 0.8 ∈1/4M and 0.98 ± 0.06, respectively. Alprenolol affected the channels in the activated and inactivated states but not in the closed states. The alprenolol-induced blockade of HERG was found to be use-dependent, exhibiting a more rapid onset and a greater steady-state block at the higher frequencies of activation. These results suggest that alprenolol prolongs the QT interval by direct inhibition of activated HERG channels.Keywords: alprenolol, HERG channel