Explainable machine learning models for early gastric cancer diagnosis.

Gastric cancer remains a significant global health concern, with a notably high incidence in East Asia. This paper explores the potential of explainable machine learning models in enhancing the early diagnosis of gastric cancer. Through comprehensive evaluations, various machine learning models, including WeightedEnsemble, CatBoost, and RandomForest, demonstrated high potential in accurately diagnosing early gastric cancer. The study emphasizes the importance of model explainability in medical diagnostics, showing how transparent, explainable models can increase trust and clinical acceptance, thereby improving diagnostic accuracy and patient outcomes. This research not only highlights key biomarkers and clinical features critical for early detection but also presents a versatile approach that could be applied to other medical diagnostics, promoting broader adoption of machine learning in clinical settings.

Unraveling the solvent stability on the cathode surface of Li-O2 batteries by using in situ vibrational spectroscopies.

In aprotic lithium-oxygen (Li-O2) batteries, solvent properties are crucial in the charge/discharge processes. Therefore, a thorough understanding of the solvent stability at the cathode surface during the oxygen reduction/evolution reactions (ORR/OER) is essential for the rational design of high-performance electrolytes. In this study, the stability of typical solvents, a series of glyme solvents with different chain lengths, has been investigated during the ORR/OER by in situ vibrational spectroscopy measurements of sum frequency generation (SFG) spectroscopy and infrared reflection absorption spectroscopy (IRRAS). The structural evolution and decomposition mechanism of the solvents during ORR/OER have been discussed based on the observations. Our results demonstrate that superoxide (O2-) generated during the ORR plays a critical role in the stability of the solvents.

Interfacial Behaviors of Giant Amphiphilic Molecules Composed of Hydrophobic Isobutyl POSS and Hydrophilic POSS Bearing Carboxylic Acid Groups at the Air-Water Interface.

The behavior of giant amphiphilic molecules at the air-water interface has become a subject of concern to researchers. Small changes in the molecular structure can cause obvious differences in the molecular arrangement and interfacial properties of the monolayer. In this study, we have systematically investigated the interfacial behaviors of the giant amphiphilic molecules with different number of hydrophobic BPOSS blocks and one hydrophilic ACPOSS block ((BPOSS)n-ACPOSS (n = 1-5)) at the air-water interface by the surface pressure-area (π-A) isotherm, Brewster angle microscopy (BAM), compression modulus measurement, and hysteresis measurement. We found that both the number of BPOSS blocks and the compression rate can significantly influence the interfacial behaviors of giant molecules. The π-A isotherms of giant molecules (BPOSS)n-ACPOSS (n = 2-5) exhibit a "cusp" phenomenon which can be attributed to the transition from monolayer to multilayer. However, the cusp is dramatically different from the "collapse" of the monolayer studied in other molecular systems, which is highly dependent on the compression rate of the monolayer. In addition, the compression modulus and hysteresis measurements reveal that the number of BPOSS blocks of (BPOSS)n-ACPOSS plays an important role in the static elasticity, stability, and reversibility of the Langmuir films.

Construction of pseudorabies virus variant attenuated vaccine: codon deoptimization of US3 and UL56 genes based on PRV gE/TK deletion strain.

Since 2011, pseudorabies based on the pseudorabies virus (PRV) variant has emerged as a serious health issue in pig farms in China. The PRV gE/TK or gE/gI/TK deletion strains protect against emerging PRV variants. However, these variants may cause lethal infections in newborn piglets without PRV antibodies. Previous studies have shown that codon deoptimization of a virulence gene causes virus attenuation. Accordingly, we deoptimized US3-S (US3 gene encoding a short isoform that represents approximately 95% of the total US3 transcription) and UL56 genes (first 10 or all codons) of PRV gE/TK deletion strain (PRVΔTK&gE-AH02) to generate six recombinant PRVs through bacterial artificial chromosome technology. In swine testicular cells, recombinant PRVs with all codon deoptimization of US3-S or UL56 genes were grown to lower titers than the parental virus. Notably, US3-S or UL56 with all codon deoptimization reduced mRNA and protein expressions. Subsequently, the safety and immunogenicity of recombinant PRVs with codon deoptimization of US3-S or UL56 are evaluated as vaccine candidates in mice and piglets. The mice inoculated with recombinant PRVs with codon deoptimization of US3-S or UL56 showed exceptional survival ability without severe clinical signs. All codons deoptimized (US3-S and UL56) significantly decreased virus load and attenuated pathological changes in the brains of the mice. Moreover, the protection efficiency offered by recombinant PRVs with codon deoptimization of US3-S or UL56 showed similar effects to PRVΔTK&gE-AH02. Remarkably, the 1-day-old PRV antibody-negative piglets inoculated with PRVΔTK&gE-US3-ST-CD (a recombinant PRV with all codon deoptimization of US3-S) presented no abnormal clinical symptoms, including fever. The piglets inoculated with PRVΔTK&gE-US3-ST-CD showed a high serum neutralization index against the PRV variant. In conclusion, these results suggest using codon deoptimization to generate innovative live attenuated PRV vaccine candidates.

BST2 negatively regulates porcine reproductive and respiratory syndrome virus replication by restricting the expression of viral proteins.

Porcine reproductive and respiratory syndrome virus (PRRSV) has seriously affected the viability of swine industries worldwide, and effective measures to control PRRSV are urgently required. Understanding the mechanisms of action of antiviral proteins is crucial for developing antiviral strategies. Interferon-induced bone marrow stromal cell antigen 2 (BST2) can inhibit the replication of various viruses via different pathways. However, little is known about the effects of BST2 on PRRSV. Therefore, this study aimed to evaluate whether the interferon-induced BST2 can inhibit PRRSV replication. We used western blotting and RT-qPCR techniques to analyze the effect of BST2 overexpression and knockdown on PRRSV replication. Overexpression of BST2 inhibited the replication of PRRSV, whereas knockdown of BST2 by small interfering RNA promoted PRRSV replication. Additionally, the expression of BST2 was upregulated during the early phase of PRRSV infection in porcine alveolar macrophages. Analysis of PRRSV proteins showed that BST2 restricted the expression of several non-structural viral proteins. BST2 downregulated the expression of Nsp12 through a proteasome-dependent pathway and downregulated the expression and transcription of E protein. These findings demonstrate the potential of BST2 as a critical regulator of PRRSV replication.

Probing the electrode-solution interfaces in rechargeable batteries by sum-frequency generation spectroscopy.

An in-depth understanding of the electrode-electrolyte interaction and electrochemical reactions at the electrode-solution interfaces in rechargeable batteries is essential to develop novel electrolytes and electrode materials with high performance. In this perspective, we highlight the advantages of the interface-specific sum-frequency generation (SFG) spectroscopy on the studies of the electrode-solution interface for the Li-ion and Li-O2 batteries. The SFG studies in probing solvent adsorption structures and solid-electrolyte interphase formation for the Li-ion battery are briefly reviewed. Recent progress on the SFG study of the oxygen reaction mechanisms and stability of the electrolyte in the Li-O2 battery is also discussed. Finally, we present the current perspective and future directions in the SFG studies on the electrode-electrolyte interfaces toward providing deeper insight into the mechanisms of discharging/charging and parasitic reactions in novel rechargeable battery systems.

Surface-Ligand "Liquid" to "Crystalline" Phase Transition Modulates the Solar H2 Production Quantum Efficiency of CdS Nanorod/Mediator/Hydrogenase Assemblies.

This study reports how the length of capping ligands on a nanocrystal surface affects its interfacial electron transfer (ET) with surrounding molecular electron acceptors, and consequently, impact the H2 production of a biotic-abiotic hybrid artificial photosynthetic system. Specifically, we study how the H2 production efficiency of a hybrid system, combining CdS nanorods (NRs), [NiFe] hydrogenase, and redox mediators (propyl-bridged 2,2'-bipyridinium, PDQ2+), depends on the alkyl chain length of mercaptocarboxylate ligands on the NR surface. We observe a minor decrease of the quantum yield for H2 production from 54 ± 6 to 43 ± 2% when varying the number of methylene units in the ligands from 2 to 7. In contrast, an abrupt decrease of the yield was observed from 43 ± 2 to 4 ± 1% when further increasing n from 7 to 11. ET studies reveal that the intrinsic ET rates from the NRs to the electron acceptor PDQ2+ are all within 108-109 s-1 regardless of the length of the capping ligands. However, the number of adsorbed PDQ2+ molecules on NR surfaces decreases dramatically when n ≥ 10, with the saturating number changing from 45 ± 5 to 0.3 ± 0.1 for n = 2 and 11, respectively. These results are not consistent with the commonly perceived exponential dependence of ET rates on the ligand length. Instead, they can be explained by the change of the accessibility of NR surfaces to electron acceptors from a disordered "liquid" phase at n < 7 to a more ordered "crystalline" phases at n > ∼7. These results highlight that the order of capping ligands is an important design parameter for further constructing nanocrystal/molecular assemblies in broad nanocrystal-based applications.

On the Coupling of Electron Transfer to Proton Transfer at Electrified Interfaces.

Many electrochemical processes are governed by the transfer of protons to the surface, which can be coupled with electron transfer; this electron transfer is in general non-integer and unknown a priori, but is required to hold the potential constant. In this study, we employ a combination of surface spectroscopic techniques and grand-canonical electronic-structure calculations in order to rigorously understand the thermodynamics of this process. Specifically, we explore the protonation/deprotonation of 4-mercaptobenzoic acid as a function of the applied potential. Using grand-canonical electronic-structure calculations, we directly infer the coupled electron transfer, which we find to be on the order of 0.1 electron per proton; experimentally, we also access this quantity via the potential-dependence of the pKa. We show a striking agreement between the potential-dependence of the measured pKa and that calculated with electronic-structure calculations. We further employ a simple electrostatics-based model to show that this slope can equivalently be interpreted to provide information on the degree of coupled electron transfer or the potential change at the point of the charged species.

Robust Binding of Disulfide-Substituted Rhenium Bipyridyl Complexes for CO2 Reduction on Gold Electrodes.

Heterogenization of homogenous catalysts on electrode surfaces provides a valuable approach for characterization of catalytic processes in operando conditions using surface selective spectroelectrochemistry methods. Ligand design plays a central role in the attachment mode and the resulting functionality of the heterogenized catalyst as determined by the orientation of the catalyst relative to the surface and the nature of specific interactions that modulate the redox properties under the heterogeneous electrode conditions. Here, we introduce new [Re(L)(CO)3Cl] catalysts for CO2 reduction with sulfur-based anchoring groups on a bipyridyl ligand, where L = 3,3'-disulfide-2,2'-bipyridine (SSbpy) and 3,3'-thio-2,2'-bipyridine (Sbpy). Spectroscopic and electrochemical analysis complemented by computational modeling at the density functional theory level identify the complex [Re(SSbpy)(CO)3Cl] as a multi-electron acceptor that combines the redox properties of both the rhenium tricarbonyl core and the disulfide functional group on the bipyridyl ligand. The first reduction at -0.85 V (vs. SCE) involves a two-electron process that breaks the disulfide bond, activating it for surface attachment. The heterogenized complex exhibits robust anchoring on gold surfaces, as probed by vibrational sum-frequency generation (SFG) spectroscopy. The binding configuration is normal to the surface, exposing the active site to the CO2 substrate in solution. The attachment mode is thus particularly suitable for electrocatalytic CO2 reduction.

Analysis of Prostate-Specific Antigen-Related Indexes, Neutrophil-to-Lymphocyte Ratio in Patients with Concurrent Benign Prostatic Hyperplasia and Histologic Prostatitis.

BACKGROUND: This study was performed to explore the total prostate-specific antigen (tPSA) concentration, free PSA (fPSA) concentration, free-to-total PSA ratio (% fPSA), tPSA density (tPSAD), and neutrophil-to-lymphocyte ratio (NLR) in blood in patients with concurrent benign prostatic hyperplasia (BPH) and histologic prostatitis, and to provide new ideas for the diagnosis of prostatitis. METHODS: Patients who underwent transurethral bipolar plasmakinetic prostatectomy from June 2017 to June 2018 were retrospectively divided into two groups according to the degree of pathological inflammation of the resected prostate tissue: group A (BPH with histologic acute and chronic inflammation), group B (BPH with histologic chronic inflammation). The preoperative PSA-related indexes and NLR in blood were respectively compared between two groups. RESULTS: Groups A and B comprised 59 and 41 cases, respectively. The values of tPSA, tPSAD, and NLR were all significantly higher in group A than B, and the value of % fPSA was significantly lower in group A than B (p < 0.05). There was no significant difference for the value of fPSA between the two groups (p > 0.05). CONCLUSIONS: Histologic acute prostatitis can cause changes of PSA-related indexes and NLR in blood, which has important clinical significance in diagnosis of prostatitis.

Heterogenized Molecular Catalysts: Vibrational Sum-Frequency Spectroscopic, Electrochemical, and Theoretical Investigations.

Rhenium and manganese bipyridyl tricarbonyl complexes have attracted intense interest for their promising applications in photocatalytic and electrocatalytic CO2 reduction in both homogeneous and heterogenized systems. To date, there have been extensive studies on immobilizing Re catalysts on solid surfaces for higher catalytic efficiency, reduced catalyst loading, and convenient product separation. However, in order for the heterogenized molecular catalysts to achieve the combination of the best aspects of homogeneous and heterogeneous catalysts, it is essential to understand the fundamental physicochemical properties of such heterogeneous systems, such as surface-bound structures of Re/Mn catalysts, substrate-adsorbate interactions, and photoinduced or electric-field-induced effects on Re/Mn catalysts. For example, the surface may act to (un)block substrates, (un)trap charges, (de)stabilize particular intermediates (and thus affect scaling relations), and shift potentials in different directions, just as protein environments do. The close collaboration between the Lian, Batista, and Kubiak groups has resulted in an integrated approach to investigate how the semiconductor or metal surface affects the properties of the attached catalyst. Synthetic strategies to achieve stable and controlled attachment of Re/Mn molecular catalysts have been developed. Steady-state, time-resolved, and electrochemical vibrational sum-frequency generation (SFG) spectroscopic studies have provided insight into the effects of interfacial structures, ultrafast vibrational energy relaxation, and electric field on the Re/Mn catalysts, respectively. Various computational methods utilizing density functional theory (DFT) have been developed and applied to determine the molecular orientation by direct comparison to spectroscopy, unravel vibrational energy relaxation mechanisms, and quantify the interfacial electric field strength of the Re/Mn catalyst systems. This Account starts with a discussion of the recent progress in determining the surface-bound structures of Re catalysts on semiconductor and Au surfaces by a combined vibrational SFG and DFT study. The effects of crystal facet, length of anchoring ligands, and doping of the semiconductor on the bound structures of Re catalysts and of the substrate itself are discussed. This is followed by a summary of the progress in understanding the vibrational relaxation (VR) dynamics of Re catalysts covalently adsorbed on semiconductor and metal surfaces. The VR processes of Re catalysts on TiO2 films and TiO2 single crystals and a Re catalyst tethered on Au, particularly the role of electron-hole pair (EHP)-induced coupling on the VR of the Re catalyst bound on Au, are discussed. The Account also summarizes recent studies in quantifying the electric field strength experienced by the catalytically active site of the Re/Mn catalyst bound on a Au electrode based on a combined electrochemical SFG and DFT study of the Stark tuning of the CO stretching modes of these catalysts. Finally, future research directions on surface-immobilized molecular catalyst systems are discussed.

CO2 Reduction Catalysts on Gold Electrode Surfaces Influenced by Large Electric Fields.

Attaching molecular catalysts to metal and semiconductor electrodes is a promising approach to developing new catalytic electrodes with combined advantages of molecular and heterogeneous catalysts. However, the effect of the interfacial electric field on the stability, activity, and selectivity of the catalysts is often poorly understood due to the complexity of interfaces. In this work, we examine the strength of the interfacial field at the binding site of CO2 reduction catalysts including Re(S-2,2'-bipyridine)(CO)3Cl and Mn(S-2,2'-bipyridine)(CO)3Br immobilized on Au electrodes. The vibrational spectra are probed by sum frequency generation spectroscopy (SFG), showing pronounced potential-dependent frequency shifts of the carbonyl stretching modes. Calculations of SFG spectra and Stark tuning rates based on density functional theory allow for direct interpretation of the configurations of the catalysts bound to the surfaces and the influence of the interfacial electric field. We find that electrocatalysts supported on Au electrodes have tilt angles of about 65-75° relative to the surface normal with one of the carbonyl ligands in direct contact with the surface. Large interfacial electric fields of 108-109 V/m are determined through the analysis of experimental frequency shifts and theoretical Stark tuning rates of the symmetric CO stretching mode. These large electric fields thus significantly influence the CO2 binding site.

Safety and immunogenicity of an attenuated Chinese pseudorabies variant by dual deletion of TK&gE genes.

BACKGROUND: Since the outbreak of a new emerging virulent pseudorabies virus mutant in Chinese pig herds, intensive research has been focused on the construction of novel gene deletion vaccine based on the variant virulent viruses. An ideal vaccine candidate is expected to have a balanced safety and immunogenicity. RESULTS: From the infectious clone of PRV AH02LA strain, a TK deletion mutant was generated through two-step Red mutagenesis. After homologous recombination with a transfer vector, a TK&gE dual deficient mutant PRV (PRVΔTK&gE-AH02) was generated, and its structure verified by PCR, RFLP and sequencing. Growth kinetics test showed that PRVΔTK&gE-AH02 reached a titer of 107.5 TCID50 /mL on ST cells. The PRVΔTK&gE-AH02 at a dose of 106.0 TCID50 /animal was not virulent in mice or 1-day-old piglets with maternal PRV antibodies. No clinical signs or virus shedding were detected in 28~ 35-day-old piglets without maternal PRV antibodies after nasal or intramuscular administration with a dose of 106.0 TCID50, although it caused one death of four 1-day-old piglets without maternal PRV antibodies. In the efficiency test of PRVΔTK&gE-AH02, all four 28~ 35-day-old piglets without PRV antibody in the challenge control showed typical clinical symptoms and virus shedding, and two died at 4~ 5 days post challenge. All piglets in 105.0, 104.0 and 103.0 TCID50/dose PRVΔTK&gE-AH02 groups provided complete protection against challenge at only 7 days post intramuscular vaccination. More importantly, PRVΔTK&gE-AH02 stopped virus shedding in these piglets. In contrast, all four piglets in PRV Bartha K61 vaccine group developed high body temperature (≥40.5 °C) and viral shedding, despite they had mild or even no clinical symptoms. CONCLUSIONS: The constructed TK&gE dual deletion mutant PRVΔTK&gE-AH02 can reach high titers on ST cells. The live vaccine of PRVΔTK&gE-AH02 is highly safe, and can not only provide clinical protection but also stops virus shedding. This study suggests that PRVΔTK&gE-AH02 might work as a promising vaccine candidate to combat the PRV variant emerging in Chinese herds since 2011.

Surface-Restructuring Differences between Polyrotaxanes and Random Copolymers in Aqueous Environment.

In the present study, we investigated the surface reorganization behaviors and adsorption conformations of fibrinogen on the surface of polyrotaxanes containing different amounts of α-cyclodextrin (α-CD) by using surface-sensitive vibrational spectroscopy sum frequency generation (SFG). For comparison, behaviors of the surface restructuring and fibrinogen adsorption on the random copolymers containing similar terminal groups were also investigated. It was found that larger amounts of BMA moieties of polyrotaxanes form ordered surface structures after immersion in water for 48 h. Furthermore, the polyrotaxane surfaces exhibit a much higher capability of fibrinogen adsorption than the random copolymer surfaces. The water-induced surface restructuring of the polyrotaxane films slightly affects the adsorption structure of the fibrinogen molecules.

Electron-Hole-Pair-Induced Vibrational Energy Relaxation of Rhenium Catalysts on Gold Surfaces.

A combination of time-resolved vibrational spectroscopy and density functional theory techniques have been applied to study the vibrational energy relaxation dynamics of the Re(4,4'-dicyano-2,2'-bipyridine)(CO)3Cl (Re(CO)3Cl) catalyst for CO2 to CO conversion bound to gold surfaces. The kinetics of vibrational relaxation exhibits a biexponential decay including an ultrafast initial relaxation and complete recovery of the ground vibrational state. Ab initio molecular dynamics simulations and time-dependent perturbation theory reveal the former to be due to vibrational population exchange between CO stretching modes and the latter to be a combination of intramolecular vibrational relaxation (IVR) and electron-hole pair (EHP)-induced energy transfer into the gold substrate. EHP-induced energy transfer from the Re(CO)3Cl adsorbate into the gold surface occurs on the same time scale as IVR of Re(CO)3Cl in aprotic solvents. Therefore, it is expected to be particularly relevant to understanding the reduced catalytic activity of the homogeneous catalyst when anchored to a metal surface.

Salt Effects on Surface Structures of Polyelectrolyte Multilayers (PEMs) Investigated by Vibrational Sum Frequency Generation (SFG) Spectroscopy.

Sum frequency generation (SFG) vibrational spectroscopy was employed to investigate the surface structures of polyelectrolyte multilayers (PEMs) constructed by sequentially alternating adsorption of poly(diallyldimethylammonium chloride) (PDDA) and poly(styrenesulfonate) (PSS). It was found that the surface structures and surface charge density of the as-deposited PEMs of PDDA/PSS significantly depend on the concentration of sodium chloride (NaCl) present in the polyelectrolyte solutions. Furthermore, it was found that the surface structure of the as-deposited PEMs is in a metastable state and will reach the equilibrium state by diffusion of the polyelectrolyte chain after an aging process, resulting in a polyelectrolyte mixture on the PEM surfaces.

Oxidative Degradation of the Monolayer of 1-Palmitoyl-2-Oleoyl-sn-Glycero-3-Phosphocholine (POPC) in Low-Level Ozone.

Ambient ozone is a common pollutant in the atmosphere that has an extremely high oxidative ability, can dramatically change the structure and functionality of biomolecules, and is harmful to public health. However, the knowledge about the influence of low-level ozone is still very limited at a molecular level. In the present study, the monolayer of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC, 16:0-18:1 PC) as well as its binary mixed monolayer with 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC, 16:0 PC), which are widely found in many biological systems, have been systematically investigated in a low-level ozone environment (20 ± 10 ppb), by π-A isotherm, sum frequency generation (SFG) vibrational spectroscopy, and atomic force microscopy (AFM). Our results demonstrate that the POPC monolayer is unstable and the C═C moieties in the oleyl chain are selectively oxidized by the low-level ozone. The oxidized lipids from POPC initially remain and reorientate the hydrophilic portion to the water surface and gradually dissolve into the aqueous solution. One should take great caution when using unsaturated lipid molecules to avoid their possible oxidation in the ambient environment. The present study expands and deepens our insights into the oxidation mechanism of unsaturated lipids at a molecular level.

Structural Reorganization and Fibrinogen Adsorption Behaviors on the Polyrotaxane Surfaces Investigated by Sum Frequency Generation Spectroscopy.

Polyrotaxanes, such as supramolecular assemblies with methylated α-cyclodextrins (α-CDs) as host molecules noncovalently threaded on the linear polymer backbone, are promising materials for biomedical applications because they allow adsorbed proteins possessing a high surface flexibility as well as control of the cellular morphology and adhesion. To provide a general design principle for biomedical materials, we examined the surface reorganization behaviors and adsorption conformations of fibrinogen on the polyrotaxane surfaces with comparison to several random copolymers by sum frequency generation (SFG) vibrational spectroscopy. We showed that the polyrotaxane (OMe-PRX-PMB) with methylated α-CDs as the host molecule exhibited unique surface structures in an aqueous environment. The hydrophobic interaction between the methoxy groups of the methylated α-CD molecules and methyl groups of the n-butyl methacrylate (BMA) side chains may dominate the surface restructuring behavior of the OMe-PRX-PMB. The orientation analysis revealed that the orientation of the fibrinogen adsorbed on the OMe-PRX-PMB surface is close to a single distribution, which is different from the adsorption behaviors of fibrinogen on other polyrotaxane or random copolymer surfaces.

Construction of a recombinant duck enteritis virus (DEV) expressing hemagglutinin of H5N1 avian influenza virus based on an infectious clone of DEV vaccine strain and evaluation of its efficacy in ducks and chickens.

BACKGROUND: Highly pathogenic avian influenza virus (AIV) subtype H5N1 remains a threat to poultry. Duck enteritis virus (DEV)-vectored vaccines expressing AIV H5N1 hemagglutinin (HA) may be viable AIV and DEV vaccine candidates. METHODS: To facilitate the generation and further improvement of DEV-vectored HA(H5) vaccines, we first constructed an infectious clone of DEV Chinese vaccine strain C-KCE (DEV(C-KCE)). Then, we generated a DEV-vectored HA(H5) vaccine (DEV-H5(UL55)) based on the bacterial artificial chromosome (BAC) by inserting a synthesized HA(H5) expression cassette with a pMCMV IE promoter and a consensus HA sequence into the noncoding area between UL55 and LORF11. The immunogenicity and protective efficacy of the resulting recombinant vaccine against DEV and AIV H5N1 were evaluated in both ducks and chickens. RESULTS: The successful construction of DEV BAC and DEV-H5(UL55) was verified by restriction fragment length polymorphism analysis. Recovered virus from the BAC or mutants showed similar growth kinetics to their parental viruses. The robust expression of HA in chicken embryo fibroblasts infected with the DEV-vectored vaccine was confirmed by indirect immunofluorescence and western blotting analyses. A single dose of 10(6) TCID50 DEV-vectored vaccine provided 100 % protection against duck viral enteritis in ducks, and the hemagglutination inhibition (HI) antibody titer of AIV H5N1 with a peak of 8.2 log2 was detected in 3-week-old layer chickens. In contrast, only very weak HI titers were observed in ducks immunized with 10(7) TCID50 DEV-vectored vaccine. A mortality rate of 60 % (6/10) was observed in 1-week-old specific pathogen free chickens inoculated with 10(6) TCID50 DEV-vectored vaccine. CONCLUSIONS: We demonstrate the following in this study. (i) The constructed BAC is a whole genome clone of DEV(C-KCE). (ii) The insertion of an HA expression cassette sequence into the noncoding area between UL55 and LORF11 of DEV(C-KCE) affects neither the growth kinetics of the virus nor its protection against DEV. (iii) DEV-H5(UL55) can generate a strong humoral immune response in 3-week-old chickens, despite the virulence of this virus observed in 1-week-old chickens. (iv) DEV-H5(UL55) induces a weak HI titer in ducks. An increase in the HI titers induced by DEV-vectored HA(H5) will be required prior to its wide application.

Molecular orientation of organic thin films on dielectric solid substrates: a phase-sensitive vibrational SFG study.

Broadband phase-sensitive vibrational sum frequency generation (SFG) spectroscopy was utilized to study the molecular orientation of molecules adsorbed on dielectric solid substrates. A gold thin film was employed to generate a SFG signal as a local oscillator (LO). To simplify the phase measurement, a self-assembled monolayer (SAM) of octadecyltrichlorosilane (OTS) was used as a standard sample for phase correction of the phase-sensitive SFG measurements on the solid/air interface. It was demonstrated that the absolute orientation of molecules in the LB films on a fused quartz surface can be clearly distinguished by phase-sensitive SFG measurement. In addition, the observation on the SAM of d35-OTS reveals that the two C-H stretching modes for α-CH2 group are in opposite phase. Furthermore, by using the present phase-sensitive SFG setup, the orientation flipping of water molecules on positively and negatively charged solid/liquid interface can be distinguished.