Viral Dynamics of Omicron and Delta Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Variants With Implications for Timing of Release from Isolation: A Longitudinal Cohort Study.

BACKGROUND: In January 2022, US guidelines shifted to recommend isolation for 5 days from symptom onset, followed by 5 days of mask-wearing. However, viral dynamics and variant and vaccination impact on culture conversion are largely unknown. METHODS: We conducted a longitudinal study on a university campus, collecting daily anterior nasal swabs for at least 10 days for reverse-transcription polymerase chain reaction (RT-PCR) testing and culture, with antigen rapid diagnostic testing (RDT) on a subset. We compared culture positivity beyond day 5, time to culture conversion, and cycle threshold trend when calculated from diagnostic test, from symptom onset, by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variant, and by vaccination status. We evaluated sensitivity and specificity of RDT on days 4-6 compared with culture. RESULTS: Among 92 SARS-CoV-2 RT-PCR-positive participants, all completed the initial vaccine series; 17 (18.5%) were infected with Delta and 75 (81.5%) with Omicron. Seventeen percent of participants had positive cultures beyond day 5 from symptom onset, with the latest on day 12. There was no difference in time to culture conversion by variant or vaccination status. For 14 substudy participants, sensitivity and specificity of day 4-6 RDT were 100% and 86%, respectively. CONCLUSIONS: The majority of our Delta- and Omicron-infected cohort culture-converted by day 6, with no further impact of booster vaccination on sterilization or cycle threshold decay. We found that rapid antigen testing may provide reassurance of lack of infectiousness, though guidance to mask for days 6-10 is supported by our finding that 17% of participants remained culture-positive after isolation.

Identification of Small Molecules with Improved Potency against Orthopoxviruses from Vaccinia to Smallpox.

The genus Orthopoxvirus contains several human pathogens, including vaccinia, monkeypox, cowpox, and variola virus, the causative agent of smallpox. Although there are a few effective vaccines, widespread prophylactic vaccination has ceased and is unlikely to resume, making therapeutics increasingly important to treat poxvirus disease. Here, we described efforts to improve the potency of the anti-poxvirus small molecule CMLDBU6128. This class of small molecules, referred to as pyridopyrimidinones (PDPMs), showed a wide range of biological activities. Through the synthesis and testing of several exploratory chemical libraries based on this molecule, we identified several compounds that had increased potency from the micromolar into the nanomolar range. Two compounds, designated (12) and (16), showed inhibitory concentrations of 326 nM and 101 nM, respectively, which was more than a 10-fold increase in potency to CMLDBU6128 with an inhibitory concentration of around 6 µM. We also expanded our investigation of the breadth of action of these molecules and showed that they can inhibit the replication of variola virus, a related orthopoxvirus. Together, these findings highlighted the promise of this new class of antipoxviral agents as broad-spectrum small molecules with significant potential to be developed as antiviral therapy. This would add a small molecule option for therapy of spreading diseases, including monkeypox and cowpox viruses, that would also be expected to have efficacy against smallpox.

Liquid Phase Oxidation of Hydrocarbons to High-Value Chemicals in Microfluidic Reactors - Prospects and Challenges.

Liquid phase oxidation (LPO) of hydrocarbon is an industrially important process to produce petrochemicals and pharmaceuticals. It follows a free radical path having initiation, propagation and termination. The initiation step is slow while the propagation and termination steps are fast. The main challenge of such process is to control product selectivity at an appreciable conversion level. With the advancement of microfluidic reactor technology, it is possible to control the free radical steps. The present contribution critically reviewed the reaction engineering aspects of LPO of hydrocarbon, the influence of microfluidic reactor design and operation on reaction mechanism, conversion and product selectivity. It also outlines the challenges associated with microfluidic reactor operation, and prospects to apply the understanding from microfluidic reactors in few sectors. The understanding from the free radical oxidation process can also be applied to any other free radical processes.

Recent Advances in Bimetallic Catalysts for Hydrogen Production from Ammonia.

The emerging concept of the hydrogen economy is facing challenges associated with hydrogen storage and transport. The utilization of ammonia as an energy (hydrogen) carrier for the on-site generation of hydrogen via ammonia decomposition has gained attraction among the scientific community. Ruthenium-based catalysts are highly active but their high cost and less abundance are limitations for scale-up application. Therefore, combining ruthenium with cheaper transition metals such as nickel, cobalt, iron, molybdenum, etc., to generate metal-metal (bimetallic) surfaces suitable for ammonia decomposition has been investigated in recent years. Herein, the recent trends in developing bimetallic catalyst systems, the role of metal type, support materials, promoter, synthesis techniques, and the investigations of the reaction kinetics and mechanism for ammonia decomposition have been reviewed.

Hybrid support vector regression and crow search algorithm for modeling and multiobjective optimization of microalgae-based wastewater treatment.

Microalgae-based wastewater treatment (and biomass production) is an environmentally benign and energetically efficient technique as compared to traditional practices. The present study is focused on optimization of the major treatment variables such as temperature, light-dark cycle (LD), and nitrogen (N)-to-phosphate (P) ratio (N/P) for the elimination of N and P from tertiary municipal wastewater utilizing Chlorella kessleri microalgae species. In this regard, a hybrid support vector regression (SVR) technique integrated with the crow search algorithm has been applied as a novel modeling/optimization tool. The SVR models were formulated using the experimental data, which were furnished according to the response surface methodology with Box-Behnken Design. Various statistical indicators, including mean absolute percentage error, Taylor diagram, and fractional bias, confirmed the superior performance of SVR models as compared to the response surface methodology (RSM) and generalized linear model (GLM). Finally, the best SVR model was hybridized with the crow search algorithm for single/multi-objective optimizations to acquire the global optimal treatment conditions for maximum N and P removal efficiencies. The best-operating conditions were found to be 29.3°C, 24/0 h/h of LD, and 6:1 of N/P, with N and P elimination efficiencies of 99.97 and 93.48%, respectively. The optimized values were further confirmed by new experimental data.

FHBC, a Hexa-peri-hexabenzocoronene-Fluorene Hybrid: A Platform for Highly Soluble, Easily Functionalizable HBCs with an Expanded Graphitic Core.

Materials based upon hexa-peri-hexabenzocoronenes (HBCs) show significant promise in a variety of photovoltaic applications. There remains the need, however, for a soluble, versatile, HBC-based platform, which can be tailored by incorporation of electroactive groups or groups that can prompt self-assembly. The synthesis of a HBC-fluorene hybrid is presented that contains an expanded graphitic core that is highly soluble, resists aggregation, and can be readily functionalized at its vertices. This new HBC platform can be tailored to incorporate six electroactive groups at its vertices, as exemplified by a facile synthesis of a representative hexaaryl derivative of FHBC. Synthesis of new FHBC derivatives, containing electroactive functional groups that can allow controlled self-assembly, may serve as potential long-range charge-transfer materials for photovoltaic applications.

Dihedral-Angle-Controlled Crossover from Static Hole Delocalization to Dynamic Hopping in Biaryl Cation Radicals.

In cases of coherent charge-transfer mechanism in biaryl compounds the rates follow a squared cosine trend with varying dihedral angle. Herein we demonstrate using a series of biaryl cation radicals with varying dihedral angles that the hole stabilization shows two different regimes where the mechanism of the hole stabilization switches over from (static) delocalization over both aryl rings to (dynamic) hopping. The experimental data and DFT calculations of biaryls with different dihedral angles unequivocally support that a crossover from delocalization to hopping occurs at a unique dihedral angle where the electronic coupling (Hab ) is one half of reorganization (λ), that is, Hab =λ/2. The implication of this finding in non-coherent charge-transfer rates is being investigated.

A search for blues brothers: X-ray crystallographic/spectroscopic characterization of the tetraarylbenzidine cation radical as a product of aging of solid magic blue.

Magic blue (MB+˙ SbCl6− salt), i.e. tris-4-bromophenylamminium cation radical, is a routinely employed one-electron oxidant that slowly decomposes in the solid state upon storage to form so called 'blues brothers', which often complicate the quantitative analyses of the oxidation processes. Herein, we disclose the identity of the main 'blues brother' as the cation radical and dication of tetrakis-(4-bromophenyl)benzidine (TAB) by a combined DFT and experimental approach, including isolation of TAB+˙ SbCl6− and its X-ray crystallography characterization. The formation of TAB in aged magic blue samples occurs by a Scholl-type coupling of a pair of MB followed by a loss of molecular bromine. The recognition of this fact led us to the rational design and synthesis of tris(2-bromo-4-tert-butylphenyl)amine, referred to as 'blues cousin' (BC: Eox1 = 0.78 V vs. Fc/Fc+, λmax(BC+˙) = 805 nm, εmax = 9930 cm−1 M−1), whose oxidative dimerization is significantly hampered by positioning the sterically demanding tert-butyl groups at the para-positions of the aryl rings. A ready two-step synthesis of BC from triphenylamine and the high stability of its cation radical (BC+˙) promise that BC will serve as a ready replacement for MB and an oxidant of choice for mechanistic investigations of one-electron transfer processes in organic, inorganic, and organometallic transformations.

Biological CO2 fixation using Chlorella vulgaris and its thermal characteristics through thermogravimetric analysis.

The present research is focused on cultivation of microalgae strain Chlorella vulgaris for bio-fixation of CO2 coupled with biomass production. In this regard, a single semi-batch vertical tubular photobioreactor and four similar photobioreactors in series have been employed. The concentration of CO2 in the feed stream was varied from 2 to 12 % (v/v) by adjusting CO2 to air ratio. The amount of CO2 capture and algae growth were monitored by measuring decrease of CO2 concentration in the gas phase, microalgal cell density, and algal biomass production rate. The results show that 4 % CO2 gives maximum amount of biomass (0.9 g L(-1)) and productivity (0.118 g L(-1) day(-1)) of C. vulgaris in a single reactor. In series reactors, average productivity per reactor found to be 0.078 g L(-1) day(-1). The maximum CO2 uptake for single reactor also found with 4 % CO2, and it is around 0.2 g L(-1) day(-1). In series reactors, average CO2 uptake is 0.13 g L(-1) day(-1) per reactor. TOC analysis shows that the carbon content of the produced biomass is around 40.67 % of total weight. The thermochemical characteristics of the cultivated C. vulgaris samples were analyzed in the presence of air. All samples burn above 200 °C and the combustion rate become faster at around 600 °C. Almost 98 wt% of the produced biomass is combustible in this range.

Application of 1D and 2D MFR reactor technology for the isolation of insecticidal and anti-microbial properties from pyrolysis bio-oils.

Valuable chemicals can be separated from agricultural residues by chemical or thermochemical processes. The application of pyrolysis has already been demonstrated as an efficient means to produce a liquid with a high concentration of desired product. The objective of this study was to apply an insect and microorganism bioassay-guided approach to separate and isolate pesticidal compounds from bio-oil produced through biomass pyrolysis. Tobacco leaf (Nicotianata bacum), tomato plant (Solanum lycopersicum), and spent coffee (Coffea arabica) grounds were pyrolyzed at 10°C/min from ambient to 565°C using the mechanically fluidized reactor (MFR). With one-dimensional (1D) MFR pyrolysis, the composition of the product vapors varied as the reactor temperature was raised allowing for the selection of the temperature range that corresponds to vapors with a high concentration of pesticidal properties. Further product separation was performed in a fractional condensation train, or 2D MFR pyrolysis, thus allowing for the separation of vapor components according to their condensation temperature. The 300-400°C tobacco and tomato bio-oil cuts from the 1D MFR showed the highest insecticidal and anti-microbial activity compared to the other bio-oil cuts. The 300-350 and 350-400°C bio-oil cuts produced by 2D MFR had the highest insecticidal activity when the bio-oil was collected from the 210°C condenser. The tobacco and tomato bio-oil had similar insecticidal activity (LC50 of 2.1 and 2.2 mg/mL) when the bio-oil was collected in the 210°C condenser from the 300-350°C reactor temperature gases. The 2D MFR does concentrate the pesticidal products compared to the 1D MFR and thus can reduce the need for further separation steps such as solvent extraction.

Self-assembly of Ni(II) with a chiral ligand pair vs. mixture of the chiral ligand pair: structural features and recognition ability of Ni2L4 cages.

The self-assembly of NiCl2 with a chiral bidentate ligand pair, (1R,2S)-(+)- and (1S,2R)-(-)-1-(nicotinamido)-2,3-dihydro-1H-inden-2-yl nicotinate (r,s-L and s,r-L) in a mixture of ethanol and dioxane, gives rise to stable crystals consisting of [2Cl@Ni2Cl2(s,r-L)4(H2O)2]·4C4H8O2·EtOH and [2Cl@Ni2Cl2(r,s-L)4(H2O)2]·4C4H8O2·EtOH chiral cages, respectively, with two encapsulated chloride anions in the cavities. The most interesting feature is that the self-assembly of NiCl2 with the mixture of r,s-L and s,r-L (1 : 1-1 : 4) produces crystals of thermodynamically stable achiral cages, [2Cl·2H2O@Ni2Cl2(s,r-L)2(r,s-L)2(H2O)2]·7C4H8O2, in the molar ratio range. Furthermore, the [2Cl@Ni2Cl2(s,r-L)4(H2O)2]·4C4H8O2·EtOH and [2Cl@Ni2Cl2(r,s-L)4(H2O)2]·4C4H8O2·EtOH chiral crystals can recognize the pairs of L-,D-tryptophan and L-,D-cysteine via cyclic voltammetry (CV) signals, in contrast to the [2Cl·2H2O@Ni2Cl2(s,r-L)2(r,s-L)2(H2O)2]·7C4H8O2 achiral crystal.

Thermocatalytic Decomposition of Methane: A Review on Carbon-Based Catalysts.

The global initiatives on sustainable and green energy resources as well as large methane reserves have encouraged more research to convert methane to hydrogen. Catalytic decomposition of methane (CDM) is one optimistic route to generate clean hydrogen and value-added carbon without the emission of harmful greenhouse gases, typically known as blue hydrogen. This Review begins with an attempt to understand fundamentals of a CDM process in terms of thermodynamics and the prerequisite characteristics of the catalyst materials. In-depth understanding of rate-determining steps of the heterogeneous catalytic reaction taking place over the catalyst surfaces is crucial for the development of novel catalysts and process conditions for a successful CDM process. The design of state-of-the-art catalysts through both computational and experimental optimizations is the need of hour, as it largely governs the economy of the process. Recent mono- and bimetallic supported and unsupported materials used in CDM process have been highlighted and classified based on their performances under specific reaction conditions, with an understanding of their advantages and limitations. Metal oxides and zeolites have shown interesting performance as support materials for Fe- and Ni-based catalysts, especially in the presence of promoters, by developing strong metal-support interactions or by enhancing the carbon diffusion rates. Carbonaceous catalysts exhibit lower conversions without metal active species and largely result in the formation of amorphous carbon. However, the stability of carbon catalysts is better than that of metal oxides at higher temperatures, and the overall performance depends on the operating conditions, catalyst properties, and reactor configurations. Although efforts to summarize the state-of-art have been reported in literature, they lack systematic analysis on the development of stable and commercially appealing CDM technology. In this work, carbon catalysts are seen as promising futuristic pathways for sustained H2 production and high yields of value-added carbon nanomaterials. The influence of the carbon source, particle size, surface area, and active sites on the activity of carbon materials as catalysts and support templates has been demonstrated. Additionally, the catalyst deactivation process has been discussed, and different regeneration techniques have been evaluated. Recent studies on theoretical models towards better performance have been summarized, and future prospects for novel CDM catalyst development have been recommended.

Viral dynamics of Omicron and Delta SARS-CoV-2 variants with implications for timing of release from isolation: a longitudinal cohort study.

Background: In January 2022, United States guidelines shifted to recommend isolation for 5 days from symptom onset, followed by 5 days of mask wearing. However, viral dynamics and variant and vaccination impact on culture conversion are largely unknown. Methods: We conducted a longitudinal study on a university campus, collecting daily anterior nasal swabs for at least 10 days for RT-PCR and culture, with antigen rapid diagnostic testing (RDT) on a subset. We compared culture positivity beyond day 5, time to culture conversion, and cycle threshold trend when calculated from diagnostic test, from symptom onset, by SARS-CoV-2 variant, and by vaccination status. We evaluated sensitivity and specificity of RDT on days 4-6 compared to culture. Results: Among 92 SARS-CoV-2 RT-PCR positive participants, all completed the initial vaccine series, 17 (18.5%) were infected with Delta and 75 (81.5%) with Omicron. Seventeen percent of participants had positive cultures beyond day 5 from symptom onset with the latest on day 12. There was no difference in time to culture conversion by variant or vaccination status. For the 14 sub-study participants, sensitivity and specificity of RDT were 100% and 86% respectively. Conclusions: The majority of our Delta- and Omicron-infected cohort culture-converted by day 6, with no further impact of booster vaccination on sterilization or cycle threshold decay. We found that rapid antigen testing may provide reassurance of lack of infectiousness, though masking for a full 10 days is necessary to prevent transmission from the 17% of individuals who remain culture positive after isolation. Main Point: Beyond day 5, 17% of our Delta and Omicron-infected cohort were culture positive. We saw no significant impact of booster vaccination on within-host Omicron viral dynamics. Additionally, we found that rapid antigen testing may provide reassurance of lack of infectiousness.

Apparent Kinetics of Co-Gasification of Biomass and Vacuum Gas Oil (VGO).

This communication reports the beneficial effects of co-gasification of biomass and residual oil to produce syngas. In this regard, various blends of glucose (a biomass surrogate) to vacuum gas oil (VGO) have been employed to investigate the synergic effects on the gasification process. The non-isothermal co-gasification experiments were conducted in a thermogravimetric analyzer at different heating rates and gasifying agents. The analysis showed that the co-gasification rate increased with the increase of glucose content in the feedstock. The presence of the oxygen in the biomass molecules helped the overall gasification process. The maximum gasification rate of 42.70 wt/min (DTGmax ) was observed with 25 wt% glucose containing sample. The use of gasifying agents appeared to have some influence, especially during high temperature gasification of the glucose-VGO blends. At a same gasification temperature, the co-gasification rate of glucose-VGO blends were found to be 125.7 wt/min and 98.59 wt%/min for N2 and CO2 , respectively. The kinetics of the co-gasification of glucose-VGO blends was conducted based on modified random pore model using TGA experimental data and implemented in MATLAB. The estimated activation energy and rate constants were found to be consistent to the observed co-gasification rates. The apparent activation energies of co-gasification of VGO/biomass blends with different weight percentages shows values ranging 60.56-48.25 kJ/mol. The kinetics analysis suggested that the addition of biomass helped to increase the reaction rate by lowering the activation energy required for accomplishing the reactions compared with petroleum carbonaceous feedstocks. The reaction rate constants isotherms are plotted to show that the k-values are exhibiting similar trends at moderate heating rates between 20 and 60 °C/min. This remark arises due to the nature of the reactions involved which are considered to be inherently similar in this range of heating rate.

Kinetics of Oxidative Cracking of n-Hexane to Light Olefins using Lattice Oxygen of a VOx /SrO-γAl2 O3 Catalyst.

The kinetics of oxidative cracking of n-hexane to light olefins using the lattice oxygen of VOx /SrO-γAl2 O3 catalysts has been investigated. Kinetic experiments were conducted in a CREC Riser Simulator (CERC: Chemical Reactor Engineering Center), which mimics fluidized bed reactors. The catalyst's performance is partly attributed to the moderate interaction between active VOx species and the SrO-γAl2 O3 support. This moderate interaction serves to control the release of lattice oxygen to curtail deep oxidation. The incorporation of basic SrO component in the support also helped to moderate the catalyst's acidity to checkmate excessive cracking. Langmuir-Hinshelwood model was applied to formulate the rate equations. The intrinsic kinetic parameters were obtained by fitting the experimental data to the kinetic model using a nonlinear regression algorithm at a 95% confidence interval, implemented in MATLAB. n-Hexane transforms to olefins at a specific reaction rate of 1.33 mol/gcat.s and activation energy of 119.2 kJ/mol. These values when compared with other duplets (i. e., ki° and EA ) for paraffins to olefins, show that indeed olefins are stable products of the oxidative conversion of n-hexane over VOx /SrO-γAl2 O3 under a fluidized bed condition. Values of activation energy for all COx formation routes indicate that intermediate paraffins are likely to be cracked to form CH4 than to be converted directly to COx . On the other hand, olefins may transform partly, and directly to COx (E9 =9.65 kJ/mol) than to form CH4 (E8 =89.1 kJ/mol) in the presence of excess lattice oxygen. Overall, olefins appear to be stable to deep oxidation due to the role of SrO in controlling the amount of lattice oxygen of the catalyst at the reaction temperature.

Preparation of a Sustainable Shape-Stabilized Phase Change Material for Thermal Energy Storage Based on Mg2+-Doped CaCO3/PEG Composites.

The properties of polyethylene glycol-6000 (PEG)/MgCaCO3, a low-cost shape-selective phase change material (ss-PCM), make it highly suitable for solar thermal applications. Nanosized porous MgO-doped CaCO3 with Mg molar concentrations of 5%, 10%, and 15% were synthesized using a hydrothermal technique. The prepared MgO-CaCO3 matrices were then impregnated with PEG to obtain PEG/MgCaCO3 as an ss-PCM. Samples identified as PEG-5MgCaCO3 (P-5-MCC), PEG-10MgCaCO3 (P-10-MCC), and PEG-15MgCaCO3 (P-15-MCC) were prepared and studied. Interestingly, P-10-MCC has the smallest particle size together with a good porous structure compared to the other two materials. The results of thermogravimetric analyses and differential scanning calorimetry indicate that the small particle size and porous structure facilitate the impregnation of approximately 69% of the PEG into the 10-MCC matrix. The latent heat and energy storage efficiency of PEG in the P-10-MCC sample are 152.5 J/g and 96.48%, respectively, which are significantly higher than those of comparable materials. Furthermore, in addition to the improvement of the thermal conductivity of the P-10-MCC, its supercooling is also reduced to some extent. The combined mesoporous and macro-porous structure of P-10-MCC is critical to retaining a large amount of PEG within the matrix, resulting in a high latent heat in the operating temperature range of 35-57 °C. The P-10MCC sample also demonstrates a high energy storage capacity (98.59%), high thermal energy storage/release rates, and exceptional shape-stabilized PCM properties.

Nurses' practice of blood transfusion in the United Arab Emirates: an observational study.

AIMS: The aim of this study was to document nurses' practice of red blood cells transfusion. BACKGROUND: In the United Arab Emirates hospitals, nurses are responsible for the administration of blood transfusions. The safety and effectiveness of the transfusion process is dependent, among others, on the knowledge and skills of nurses who perform the procedure. Poor practice may result in avoidable complications that may threaten patients' safety. Published work indicated that nurses' practice varied across contexts and highlighted that patients received suboptimal care and incorrect transfusion that culminated in death or morbidity. In the United Arab Emirates, publications related to nurses' practice of blood transfusion are lacking. DESIGN: Descriptive. METHODS: Data were collected by means of non-participant structured observation. Data collection was undertaken in two general public hospitals in the Emirate of Abu Dhabi, United Arab Emirates. A random sample of 50 nurses from both hospitals was selected. Each nurse was observed once, from 10 minutes prior to blood collection until 15 minutes after initiating a transfusion. RESULTS: Forty-nine nurses (98%) were observed. The maximum obtained score was 13 points of a possible score of 21, and 75% of nurses scored below the 50% level. Practice deficiencies included improper patient identification, suboptimal vital signs documentation and invalid methods of blood warming. CONCLUSIONS: Patients in both hospitals were at risk of receiving incorrect blood, suffering unobserved transfusion reaction and acquiring bacterial infection. RELEVANCE TO CLINICAL PRACTICE: This study revealed inadequate practices that nurses and hospitals should strive to change to provide a safer and more effective care that would, hopefully, minimise the risks and maximise the benefits of blood transfusion. These findings also have implications for clinical supervision and nurse education.

Chest radiograph reading panel performance in a Bangladesh pneumococcal vaccine effectiveness study.

Introduction: To evaluate WHO chest radiograph interpretation processes during a pneumococcal vaccine effectiveness study of children aged 3-35 months with suspected pneumonia in Sylhet, Bangladesh. Methods: Eight physicians masked to all data were standardised to WHO methodology and interpreted chest radiographs between 2015 and 2017. Each radiograph was randomly assigned to two primary readers. If the primary readers were discordant for image interpretability or the presence or absence of primary endpoint pneumonia (PEP), then another randomly selected, masked reader adjudicated the image (arbitrator). If the arbitrator disagreed with both primary readers, or concluded no PEP, then a masked expert reader finalised the interpretation. The expert reader also conducted blinded quality control (QC) for 20% of randomly selected images. We evaluated agreement between primary readers and between the expert QC reading and the final panel interpretation using per cent agreement, unadjusted Cohen's kappa, and a prevalence and bias-adjusted kappa. Results: Among 9723 images, the panel classified 21.3% as PEP, 77.6% no PEP and 1.1% uninterpretable. Two primary readers agreed on interpretability for 98% of images (kappa, 0.25; prevalence and bias-adjusted kappa, 0.97). Among interpretable radiographs, primary readers agreed on the presence or absence of PEP in 79% of images (kappa, 0.35; adjusted kappa, 0.57). Expert QC readings agreed with final panel conclusions on the presence or absence of PEP for 92.9% of 1652 interpretable images (kappa, 0.75; adjusted kappa, 0.85). Conclusion: Primary reader performance and QC results suggest the panel effectively applied the WHO chest radiograph criteria for pneumonia.

Replicon Particle Expressing the E2 Glycoprotein of Bovine Viral Diarrhea Virus Immunization and Evaluation of Antibody Response.

The objective of this study was to develop a new antigen delivery system using an alphavirus replicon particle (RP) to induce humoral antibody responses against bovine viral diarrhea virus (BVDV) recombinant antigen produced from envelope glycoprotein E2. An alphavirus RP expressing the E2 glycoprotein of BVDV was used for immunization of pigs. A fluorescent microsphere immunoassay (FMIA) has been applied to detect BVDV E2 antigen-specific antibody isotype in pig immunized with alphavirus RP. Full-length BVDV E2 (aa 1-375) was cleaved into several pieces, eight E2 DNA fragments, including full-length DNA, were cloned into expression vector pHUE, and the recombinant proteins expressed in BL-21 (DE3) Escherichia coli. After successful conjugation of purified proteins with microsphere beads, a multiplex FMIA platform was constructed, and BVDV E2 alphavirus-based RP-immunized animal serum samples were tested in the presence of bead-bound antigen targets. The results were represented as mean fluorescence intensity (MFI); the MFI values were converted to sample value/positive value (S/P) ratios. BVDV E2 (aa 1-183) showed the highest MFI values of eight recombinant E2 fragments when the specific activity of each fragment was tested. In immunized animals, data for BVDV E2-specific IgA, IgG, and IgM in serum and only IgG and IgA in oral fluids were recorded. The MFI values for the positive serum sample showed a 100-fold increase compared with the negative serum sample. Antibody isotype to BVDV E2 antigens showed that IgG > IgM > IgA in serum, whereas IgG > IgA > IgM in oral fluids. The data presented in this study suggested that boosting with the same doses of alphavirus RP in 3-week intervals may potentially enhance antibody response. The experimental results demonstrate that alphavirus RP-expressing BVDV E2 antigen induces antibody response in pig.

Multiplex Detection of IgG and IgM to Rift Valley Fever Virus Nucleoprotein, Nonstructural Proteins, and Glycoprotein in Ovine and Bovine.

A multiplex fluorescence microsphere immunoassay (FMIA) was used to detect bovine and ovine IgM and IgG antibodies to several Rift Valley fever virus (RVFV) proteins, including the major surface glycoprotein, Gn; the nonstructural proteins, NSs and NSm; and the nucleoprotein, N. Target antigens were assembled into a multiplex and tested in serum samples from infected wild-type RVFV or MP12, a modified live virus vaccine. As expected, the N protein was immunodominant and the best target for early detection of infection. Antibody activity against the other targets was also detected. The experimental results demonstrate the capabilities of FMIA for the detection of antibodies to RVFV structural and nonstructural proteins, which can be applied to future development and validation of diagnostic tests that can be used to differentiate vaccinated from infected animals.