Two-Electron-Induced Reorganization of Cobalt Coordination and Metal-Ligand Cooperative Redox Shifting Co(I) Reactivity toward CO2 Reduction.

Electrochemical reorganization of complex structures is directly related to catalytic reactivity; thus, the geometric changes of catalysts induced by electron transfer should be considered to scrutinize the reaction mechanism. Herein, we studied electron-induced reorganization patterns of six-coordinate Co complexes with neutral N-donor ligands. Upon two-electron transfer into a Co center enclosed within a bulky π-acceptor ligand, the catalytic site exhibited different reorganization patterns depending on the ligand characteristics. While a bipyridyl ligand released Co-bound solvent (CH3CN) to open a reaction site, a phenanthroline ligand caused Co-Narm (side "arm" of NNN-ligand) bond dissociation. The first electron transfer occurred in the Co(II/I) reduction step and the second electron entered the bulky π-acceptor, of which redox steps were assigned from cyclic voltammograms, magnetic moment measurements, and DFT calculations. In comparison, the Co complex of [NNNNCH3-Co(CH3CN)3](PF6)2 ([1-(CH3CN)3](PF6)2) showed a high H2 evolution reactivity (HER), whereas a series of Co complexes with bulky π-acceptors such as [NNNNCH3-Co(L)(CH3CN)](PF6)2 (L = phen ([2-CH3CN](PF6)2), bpy ([3-CH3CN](PF6)2), [NNNNCH3-Co(tpy)](PF6)2 ([4](PF6)2), and [NNNCH2-Co(phen)(CH3CN)](PF6)2 ([5-CH3CN](PF6)2)) suppressed the HER but rather enhanced the CO2 reduction reaction. The metal-ligand cooperative redox steps enabled the shift of Co(I) reactivity toward CO2 reduction. Additionally, the amine pendant attached to the NNNNCH3-ligand could stabilize the CO2 reduction intermediate through the hydrogen-bonding interaction with the Co-CO2H adduct.

Propagation of random excitations in the polar representation of strongly nonlinear four-wave systems of turbulence.

The present work is concerned with the uncertainty propagation of the wave turbulent system. In particular, we study the temporal development and long-term behavior of the probability with respect to the amplitude and phase of complex-valued waves constituting the generic four-wave system of turbulence. Our approach to approximating the target distribution function is via the three steps: (i) to grasp the physical process described by the true turbulence model as random process, (ii) to determine the stochastic differential equation whose solution exhibits statistically similar behavior with the underlying turbulent signal, and (iii) to solve the corresponding Kolmogorov forward equation. Our implementation of the methodology is distinguished by employing a number of simplified stochastic models and applying one of them in the adaptive fashion which varies subject to the different parameter regime of the true dynamical system model. Accordingly, we become able to demonstrate the effectiveness of this reduced-order modeling framework for the analysis of the turbulent system characterized by not only weak but strong interactions among the nonlinear waves. We numerically corroborate our theoretical predictions in the context of the generalized Majda-Mclaughlin-Tabak wave turbulence prototype.

Quasi-Markovian property of strong wave turbulence.

This paper is concerned with the reduced-order modeling of the strongly nonlinear wave turbulence system. The motivation for such an attempt comes from the utility of the probabilistic coarse-grained model in facilitating the theoretical and numerical analysis of the true dynamical system model. One typical practice of simplifying the complex physical model is, in the spirit of Brownian motion, to replace the nonlinear interactions by white noise forcing and linear dissipation. For the case of slowly varying longwave, the resulting Markov process is an accurate approximate model. However, this conventional scheme is highly inappropriate for the description of shortwaves because the rapidly varying turbulent signal acquires a significantly non-Markovian character resulting from the poor timescale separation between the relevant mode and the environmental wave field. To resolve the issue, we discuss a simplification technique for which the central concept is the quasi-Markovian property; a non-Markov stochastic process is referred to as quasi-Markovian if it can be represented as a component of Markovian system made by adding a finite number of auxiliary variables. Our contribution in this work is to single out the nontrivial and near resonances from the nonlinear interactions in search of the auxiliary variable. We perform a comparison analysis of the autocorrelation matrices of the true and approximate models, and numerically demonstrate the effectiveness of our Markovian formulation of the inherently non-Markov turbulent signal.

Solid-state colorimetric polydiacetylene liposome biosensor sensitized by gold nanoparticles.

Polydiacetylene (PDA), a conjugated polymer, has attracted attention for realization of a label-free real-time colorimetric biosensor because it exhibits large and rapid colorimetric responses upon the binding of biomolecules. This is due to the conformational distortion of its conjugated backbone. However, solid-state PDA biosensors for point-of-care diagnosis remain unexplored. We describe a highly sensitive solid-state biosensor based on PDA liposomes. We employed gold nanoparticles (AuNPs) on PDA liposomes as the molecular-binding-signal sensitizer, which provides additional conformational distortion in the backbone structure of PDA by exerting steric repulsion to the attached biomolecules. To prove the concept, AuNPs and a thrombin-binding-aptamer were individually functionalized on PDA liposomes, which were attached to a substrate for the detection of thrombin. We found that the sensitivity was enhanced 2.5 times in the presence of AuNPs compared with the case without AuNPs. Because the steric repulsion of the AuNPs is target-independent, we believe that our solid-state biosensor provides a path toward advanced solid-state biosensors.

Generalized Langevin equation and the linear regression model with memory.

Because the existence of cheap emulators can give rise to potentially drastic computational savings as compared to direct numerical simulations of complex turbulence models, the study of reduced models has good practical relevance, and the guidelines regarding their construction from the true dynamical system model are much in demand. In accordance with this contemporary trend in science and engineering, we provide a new approach for the rigorous derivation of the linear reduced model with memory from the one-dimensional Majda-McLaughlin-Tabak (MMT) prototypical wave turbulence in thermal equilibrium. The basic idea in obtaining the physics-constrained autoregressive model is to perform the discretization in time of the generalized Langevin equation (GLE) governing a single wave profile of the true dynamical system model; the GLE formalized in the Mori-Zwanzig (MZ) projection theory is the exact reduced-order equation and the closed-form rearrangement of the canonical equation of motion for the Hamiltonian system. We study the performance of the resulting linear non-Markovian model in addressing two different manners of uncertainty quantification (UQ) problems: prediction and filtering. In doing so, we perform a comparison analysis against the linear and nonlinear Markovian models describing the MMT system, which are also built upon the GLE under the MZ framework. Finally we discuss an optimal selection of the statistical model in applying the reduced-model approach to the UQ of the turbulent signal.

Cascade model of wave turbulence.

We propose a cascade model of wave turbulence designed to simplify the study of this phenomenon in the way that shell models simplify the study of Navier-Stokes turbulence. The model consists of resonant quartets, in which some modes are driven and damped and others are shared by pairs of quartets and transferring energy between them, mimicking the natural energy transfer mechanism in weakly turbulent waves. A set of detailed-balance conditions singles out the case of the cascade model in equilibrium, for which we can explicitly derive a Gaussian equilibrium measure and a maximum-entropy principle using a Kolmogorov forward equation. Away from equilibrium, we can approximate the second-moment dynamics of the mode amplitudes using kinetic equations. In a nonequilibrium steady state, we can also approximate the higher moments of the driven-damped mode amplitudes and characterize the distribution of the shared-mode amplitudes as Gaussian. For this latter distribution, we find an information-theoretic argument, akin to entropy maximization, which lets us conclude that arbitrary initial shared-mode amplitude distributions approach Gaussian form in forward time. The cascade model may provide insight into mechanisms governing weakly turbulent wave systems and perhaps afford computational savings as compared to direct numerical simulations of the corresponding wavelike equations.

Development of a job rotation scheduling algorithm for minimizing accumulated work load per body parts.

BACKGROUND: For the successful implementation of job rotation, jobs should be scheduled systematically so that physical workload is evenly distributed with the use of various body parts. However, while the potential benefits are widely recognized by research and industry, there is still a need for a more effective and efficient algorithm that considers multiple work-related factors in job rotation scheduling. OBJECTIVE: This study suggests a type of job rotation algorithm that aims to minimize musculoskeletal disorders with the approach of decreasing the overall workload. METHODS: Multiple work characteristics are evaluated as inputs to the proposed algorithm. Important factors, such as physical workload on specific body parts, working height, involvement of heavy lifting, and worker characteristics such as physical disorders, are included in the algorithm. For evaluation of the overall workload in a given workplace, an objective function was defined to aggregate the scores from the individual factors. A case study, where the algorithm was applied at a workplace, is presented with an examination on its applicability and effectiveness. RESULTS: With the application of the suggested algorithm in case study, the value of the final objective function, which is the weighted sum of the workload in various body parts, decreased by 71.7% when compared to a typical sequential assignment and by 84.9% when compared to a single job assignment, which is doing one job all day. CONCLUSIONS: An algorithm was developed using the data from the ergonomic evaluation tool used in the plant and from the known factors related to workload. The algorithm was developed so that it can be efficiently applied with a small amount of required inputs, while covering a wide range of work-related factors. A case study showed that the algorithm was beneficial in determining a job rotation schedule aimed at minimizing workload across body parts.

Supplementation of dietary germanium biotite enhances induction of the immune responses by foot-and-mouth disease vaccine in cattle.

BACKGROUND: After the recent outbreak of foot-and-mouth disease (FMD) in Korea, a vaccination policy has been applied to control the disease. In addition, several non-specific immune stimulators have been used without any scientific evidence that they would enhance the immune response after FMD vaccination and/or protect against FMD. Based on the current situation, the aim of this study was to evaluate the effect of the non-specific immune stimulator germanium biotite on FMD vaccination and immune responses in cattle. To achieve our goal, immune responses to FMD vaccination, such as levels of IgG and IgA, antibody duration, and virus-neutralizing titers were investigated after germanium biotite feeding. The PBMC typing and proliferative response after stimulation with mitogens, the cytokines expression level of PBMC, and the lysozyme activity in the serum were measured to evaluate the immune enhancing effects of germanium biotite following its administration. RESULTS: Following the first vaccination, high level of IgG (at 4 weeks) and IgA (at 2 and 31 weeks) titers in serum and saliva were observed in the germanium biotite-feeding group (p < 0.05). The germanium biotite group also showed high and longstanding inhibition percentage value in ELISA assay at 31 weeks (p < 0.05). Generally, higher virus-neutralizing antibody titers were observed in the feeding group at 20 and 31 weeks after vaccination. Following the feeding germanium biotite, the germanium biotite group showed increased subpopulation of CD4+ lymphocytes and MHC I+II+ cells in PBMCs at 23 week, responding to stimulation of ConA. The levels of IFN-γ (at 3 and 8 weeks), IL-1α (at 3, 11, and 23 weeks), IL-1ß (at 3, 8, and 11 weeks), and IL-4 (at 8 and 11 weeks) gene expression were also significantly increased in the feeding group (p < 0.01 and p < 0.05). Feeding with germanium biotite increased the lymphocytes' proliferative response to the stimulation of ConA and LPS at 23 weeks and lysozyme activity at 9 weeks after feeding. CONCLUSIONS: These results suggest that germanium biotite feeding could increase the protection against FMD virus infection via the induction of higher humoral and cellular immune responses in cattle.

Induction of immune responses in mice and pigs by oral administration of classical swine fever virus E2 protein expressed in rice calli.

Classical swine fever (CSF), caused by the CSF virus (CSFV), is a highly contagious disease in pigs. In Korea, vaccination using a live-attenuated strain (LOM strain) has been used to control the disease. However, parenteral vaccination using a live-attenuated strain still faces a number of problems related to storage, cost, injection stress, and differentiation of CSFV infected and vaccinated pigs. Therefore, two kinds of new candidates for oral vaccination have been developed based on the translation of the E2 gene of the SW03 strain, which was isolated from an outbreak of CSF in 2002 in Korea, in transgenic rice calli (TRCs) from Oriza sativa L. cv. Dongjin to express a recombinant E2 protein (rE2-TRCs). The expression of the recombinant E2 protein (rE2) in rE2-TRCs was confirmed using Northern blot, SDS-PAGE, and Western blot analysis. Immune responses to the rE2-TRC in mice and pigs were investigated after oral administration. The administration of rE2-TRCs increased E2-specific antibodies titers and antibody-secreting cells when compared to animals receiving the vector alone (p < 0.05 and p < 0.01). In addition, mice receiving rE2-TRCs had a higher level of CD8+ lymphocytes and Th1 cytokine immune responses to purified rE2 (prE2) in vitro than the controls (p < 0.05 and p < 0.01). Pigs receiving rE2-TRCs also showed an increase in IL-8, CCL2, and the CD8+ subpopulation in response to stimulation with prE2. These results suggest that oral administration of rE2-TRCs can induce E2-specific immune responses.

Oral immunization of mice with Saccharomyces cerevisiae expressing a neutralizing epitope of ApxIIA exotoxin from Actinobacillus pleuropneumoniae induces systemic and mucosal immune responses.

An oral delivery system based on ApxIIA#5-expressed on Saccharomyces cerevisiae was studied for its potential to induce immune responses in mice. Murine bone marrow-derived dendritic cells (DCs) stimulated in vitro with ApxIIA#5-expressed on S. cerevisiae upregulated the expression of maturation and activation markers, leading to production of tumor necrosis factor-α, interleukin (IL)-1ß, IL-12p70 and IL-10. Presentation of these activated DCs to cluster of differentiation CD4+ T cells collected from mice that had been orally immunized with the ApxIIA#5-expressed on S. cerevisiae elicited specific T-cell proliferation. In addition, the orally immunized mice had stronger antigen-specific serum IgG and IgA antibody responses and larger numbers of antigen-specific IgG and IgA antibody-secreting cells in their spleens, Peyer's patches and lamina propria than did those immunized with vector-only S. cerevisiae or those not immunized. Furthermore, oral immunization induced T helper 1-type immune responses mediated via increased serum concentrations of IgG2a and an increase predominantly of IFN-γ-producing cells in their spleens and lamina propria. Our findings suggest that surface-displayed ApxIIA#5-expressed on S. cerevisiae may be a promising candidate for an oral vaccine delivery system for eliciting systemic and mucosal immunity.

Generation of dispersion in nondispersive nonlinear waves in thermal equilibrium.

In this work, we examine the important theoretical question of whether dispersion relations can arise from purely nonlinear interactions among waves that possess no linear dispersive characteristics. Using two prototypical examples of nondispersive waves, we demonstrate how nonlinear interactions can indeed give rise to effective dispersive-wave-like characteristics in thermal equilibrium. Physically, these example systems correspond to the strong nonlinear coupling limit in the theory of wave turbulence. We derive the form of the corresponding dispersion relation, which describes the effective dispersive structures, using the generalized Langevin equations obtained in the Zwanzig-Mori projection framework. We confirm the validity of this effective dispersion relation in our numerical study using the wavenumber-frequency spectral analysis. Our work may provide insight into an important connection between highly nonlinear turbulent wave systems, possibly with no discernible dispersive properties, and the dispersive nature of the corresponding renormalized waves.

Induction of protective immune responses against challenge of Actinobacillus pleuropneumoniae by oral administration with Saccharomyces cerevisiae expressing Apx toxins in pigs.

Actinobacillus pleuropneumoniae is a causative agent of porcine pleuropneumonia, a highly contagious endemic disease of pigs worldwide, inducing significant economic losses worldwide. Apx toxins, which are correlated with the virulence of A. pleuropneumoniae, were expressed in Saccharomyces cerevisiae and its possible use as an oral vaccine has been confirmed in our previous studies using a murine model. The present study was undertaken to test the hypothesis that oral immunization using S. cerevisiae expressing either ApxI or ApxII could protect pigs against A. pleuropneumoniae as an effective way of inducing both mucosal and systemic immune responses. The surface-displayed ApxIIA#5 expressing S. cerevisiae was selected as an oral vaccine candidate by finding on induction of higher immune responses in mice after oral vaccination. The surface-displayed ApxIIA#5 expressing S. cerevisiae and the ApxIA expressing S. cerevisiae were developed to serve as an oral vaccine in pigs. The vaccinated pigs showed higher specific IgG- and IgA-related antibody activities than the non-treated control and vector control pigs. Additionally, the induced immune responses were found to protect pigs infected with A. pleuropneumoniae according to the analysis of clinical signs and the gross and microscopic pulmonary lesions. These results suggested that the surface-displayed ApxIIA#5 and ApxIA in S. cerevisiae might be a potential oral vaccine to protect pigs against porcine pleuropneumonia. Thus the present study is expected to contribute to the development of a live oral vaccine against porcine pleuropneumonia as an alternative to current conventional vaccines.

Efficacy of thiolated eudragit microspheres as an oral vaccine delivery system to induce mucosal immunity against enterotoxigenic Escherichia coli in mice.

A vaccine delivery system based on thiolated eudragit microsphere (TEMS) was studied in vivo for its ability to elicit mucosal immunity against enterotoxigenic Escherichia coli (ETEC). Groups of mice were orally immunized with F4 or F18 fimbriae of ETEC and F4 or F18 loaded in TEMS. Mice that were orally administered with F4 or F18 loaded TEMS showed higher antigen-specific IgG antibody responses in serum and antigen-specific IgA in saliva and feces than mice that were immunized with antigens only. In addition, oral vaccination of F4 or F18 loaded TEMS resulted in higher numbers of IgG and IgA antigen-specific antibody secreting cells in the spleen, lamina propria, and Peyer's patches of immunized mice than other groups. Moreover, TEMS administration loaded with F4 or F18 induced mixed Th1 and Th2 type responses based on similarly increased levels of IgG1 and IgG2a. These results suggest that F4 or F18 loaded TEMS may be a promising candidate for an oral vaccine delivery system to elicit systemic and mucosal immunity against ETEC.

Generation of transgenic corn-derived Actinobacillus pleuropneumoniae ApxIIA fused with the cholera toxin B subunit as a vaccine candidate.

Corn, one of the most important forage crops worldwide, has proven to be a useful expression vehicle due to the availability of established transformation procedures for this well-studied plant. The exotoxin Apx, a major virulence factor, is recognized as a common antigen of Actinobacillus (A.) pleuropneumoniae, the causative agent of porcine pleuropneumonia. In this study, a cholera toxin B (CTB)-ApxIIA#5 fusion protein and full-size ApxIIA expressed in corn seed, as a subunit vaccine candidate, were observed to induce Apx-specific immune responses in mice. These results suggest that transgenic corn-derived ApxIIA and CTB-ApxIIA#5 proteins are potential vaccine candidates against A. pleuropneumoniae infection.

An immunosorbent assay based on the recombinant ApxIa, ApxIIa, and ApxIIIa toxins of Actinobacillus pleuropneumoniae and its application to field sera.

Actinobacillus pleuropneumoniae is the etiologic agent of porcine pleuropneumonia, a highly contagious pulmonary disease in pigs with major economic losses for pig producers worldwide. Whereas A. pleuropneumoniae isolates are divided into 15 serotypes, the isolates secrete 4 types of exotoxins (ApxI, ApxII, ApxIII, and ApxIV), which are known as major virulence factors. In the current study, the ApxIA, ApxIIA, and ApxIIIA genes were amplified and their recombinant proteins expressed in Escherichia coli M15 cells. The antigenicity of each recombinant protein was demonstrated by Western blot and enzyme-linked immunosorbent assay (ELISA) using sera from pigs vaccinated with a subunit vaccine. When ELISAs using the recombinant antigens were optimized and then applied to sera from 320 randomized pigs in Korea, an observed increase in seroprevalence was found among sows in comparison with weaned piglets and growing pigs, indicating an age-dependent seroprevalence. The results obtained in the study suggest that the developed ELISAs may be useful for A. pleuropneumoniae vaccination strategy as a screening tool for pig herds as well as for detection of specific antibodies to Apx exotoxins.

Predicting genetic traits and epitope analysis of apxIVA in Actinobacillus pleuropneumoniae.

Actinobacillus pleuropneumoniae causes a severe hemorrhagic pneumonia in pigs. Fifteen serotypes of A. pleuropneumoniae express four different Apx toxins that belong to the pore-forming repeats-in-toxin (RTX) group of toxins. ApxIV, which is conserved and up-regulated in vivo, could be an excellent candidate for the development of a protective cross-serotype immunity vaccine, and could aid in the differential diagnosis of diseases caused by A. pleuropneumoniae. We identified and sequenced apxIVA from A. pleuropneumoniae serotype 2 isolated in Korea (Kor-ApxIVA). The Kor-ApxIVA was closely related to Switzerland (AF021919), China (CP000687), and China (GQ332268), showing 98.6%, 98.4%, and 97.2% amino acid homology, respectively. The level of amino acid homology, however, was higher than the nucleotide homology. The structural characteristics of ApxIVA showed RTX proteins, including N-terminal hydrophobic domains, signature sequences for potential acylation sites, and repeated glycine-rich nonapeptides in the C-terminal region of the protein. Thirty glycine-rich nonapeptides with the consensus sequence, L/V-X-G-G-X-G-N/D-D-X, were found in the C-terminus of the Kor-ApxIVA. In addition, the Kor-ApxIVA was predicted for the linear B-cell epitopes and conserved domains with determined peptide sequences. This genetic analysis of the Kor-ApxIVA might be an important foundation for future biological and functional research on ApxIVA.

Induction of Th1 polarized immune responses by thiolated Eudragit-coated F4 and F18 fimbriae of enterotoxigenic Escherichia coli.

Diarrhea in newborn and weaned piglets is mainly induced by enterotoxigenic Escherichia coli (ETEC) with fimbriae F4 (K88) and F18 (F107). In this study, we evaluated F4 and F18 coated with thiolated Eudragit microspheres (TEMS) as a candidate for an oral vaccine. The average particle sizes of TEMS, F4-loaded TEMS, and F18-loaded TEMS were measured as 4.2±0.75 µm, 4.7±0.50 µm, and 4.5±0.37 µm, respectively. F4 is more efficiently encapsulated than F18 in the loading with TEMS. In the release test, F4 and F18 fimbriae were protected in acidic circumstances, whereas most were released at pH 7.4 of intestine circumstances. Production of TNF-α and NO from RAW 264.7 cells was increased in a time-dependent manner after exposure to all groups, whereas only F4- or F18-loaded TEMS-stimulated IL-6 secretion. The levels of IFN-γ from mouse splenocytes after exposure to F4 or F18 were increased while IL-4 was not detectable. These results suggest that F4- and F18-loaded TEMS may effectively induce immune response with the efficient release of antigens to appropriate target sites.

Improved sero-monitoring assay for classical swine fever (CSF) using the recombinant E2 protein of a recent Korean isolate.

Classical swine fever (CSF) is a highly contagious disease of pigs that causes fever, diarrhea and paralysis, often resulting in death. E2 is the major structural protein of the CSF virus (CSFV) and mediates the entrance of the virus, subsequently inducing a neutralizing immune response. In this study, the E2 gene of a recent Korean isolate of CSF, SW03, was cloned and the DNA sequence was compared to other strains via phylogenetic analysis. With the purified E2 protein, an enzyme-linked immunosorbent assay (ELISA) was developed for the serodiagnosis of CSFV infection. The sensitivity and specificity of the E2-ELISA were 96.1% and 94.8%, respectively. A total of 17 out of 485 field-collected pig sera tested demonstrated conflicting results between two ELISA methods, a commercial kit and the E2-ELISA. Of these sera, 60% were determined to be CSFV positive by a virus neutralization test (VNT), suggesting involvement of different immune responses in the cases of CSFV infection. As the E2-ELISA was developed using a recent Korean isolate, SW03, this assay is capable of rapidly identifying newly emerging CSFV strains.

Comparative study of gamma interferon production in mice immunized with outer membrane proteins and whole bacteria of Brucella abortus.

Brucella abortus is the intracellular bacterium that causes bovine brucellosis and a chronic human disease known as undulant fever. Interferon (IFN)-gamma plays critical roles in defending against intracellular bacterial infection. In this experiment, we demonstrated the difference in IFN-gamma production between the splenocytes of mice inoculated with outer membrane proteins (OMPs) of B. abortus and whole live bacteria. Our results showed that the OMP-inoculated group showed more IFN-gamma production than did the bacteria-infected group, suggesting that OMPs are candidates for the induction of immune response.

Renormalized resonance quartets in dispersive wave turbulence.

Using the (1+1)D Majda-McLaughlin-Tabak model as an example, we present an extension of the wave turbulence (WT) theory to systems with strong nonlinearities. We demonstrate that nonlinear wave interactions renormalize the dynamics, leading to (i) a possible destruction of scaling structures in the bare wave systems and a drastic deformation of the resonant manifold even at weak nonlinearities, and (ii) creation of nonlinear resonance quartets in wave systems for which there would be no resonances as predicted by the linear dispersion relation. Finally, we derive an effective WT kinetic equation and show that our prediction of the renormalized Rayleigh-Jeans distribution is in excellent agreement with the simulation of the full wave system in equilibrium.