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Purpose Information on incentives for COVID-19 testing is needed to understand effective practices that encourage testing uptake. We describe characteristics of those who received an incentive after performing a rapid antigen test. Design Cross-sectional descriptive analysis of survey data. Setting During April 29–May 9, 2021, COVID-19 rapid antigen testing was offered in 2 Maryland cities. Sample Convenience sample of 553 adults (≥18 years) who tested and received an incentive; 93% consented to survey. Measures Survey questions assessed reasons for testing, testing history, barriers, and demographics. Analysis Robust Poisson regressions were used to determine characteristic differences based on testing history and between participants who would re-test in the future without an incentive vs participants who would not. Results The most common reasons for testing were the desire to be tested (n = 280; 54%) and convenience of location (n = 146; 28%). Those motivated by an incentive to test (n = 110; 21%) were 5.83 times as likely to state they would not test again without an incentive, compared to those with other reasons for testing (95% CI: 2.67-12.72, P < .001). Critical Limitations No comparative study group. Conclusion Results indicate internal motivation and convenience were prominent factors supporting testing uptake. Incentives may increase community testing participation, particularly among people who have never tested. Keywords COVID-19, pandemic, incentives, health behavior, community testing.

Immediately following the March 13, 2020 declaration of COVID-19 as a national emergency (1), the U.S. government began implementing national testing programs for epidemiologic surveillance, monitoring of frontline workers and populations at higher risk for acquiring COVID-19, and identifying and allocating limited testing resources. Effective testing supports identification of COVID-19 cases; facilitates isolation, quarantine, and timely treatment measures that limit the spread of SARS-CoV-2 (the virus that causes COVID-19); and guides public health officials about the incidence of COVID-19 in a community. A White House Joint Task Force, co-led by the Department of Health and Human Services (HHS) and the Federal Emergency Management Agency (FEMA), created the Community-Based Testing Sites (CBTS) program working with state and local partners (2). This report describes the timeline, services delivered, and scope of the CBTS program. During March 19, 2020-April 11, 2021, the CBTS program conducted 11,661,923 SARS-CoV-2 tests at 8,319 locations across the United States and its territories, including 402,223 (3.5%) administered through Drive-Through Testing, 10,129,142 (86.9%) through Pharmacies+ Testing, and 1,130,558 (9.7%) through Surge Testing programs. Tests administered through the CBTS program yielded 1,176,959 (10.1%) positive results for SARS-CoV-2. Among tested persons with available race data,* positive test results were highest among American Indian or Alaska Native (14.1%) and Black persons (10.4%) and lowest among White persons (9.9%), Asian persons (7.3%), and Native Hawaiian or Other Pacific Islanders (6.4%). Among persons with reported ethnicity, 25.3% were Hispanic, 15.9% of whom received a positive test result. Overall, 82.0% of test results were returned within 2 days, but the percentage of test results returned within 2 days was as low as 40.7% in July 2020 and 59.3% in December 2020 during peak testing periods. Strong partnerships enabled a rapid coordinated response to establish the federally supported CBTS program to improve access to no-charge diagnostic testing, including for frontline workers, symptomatic persons and close contacts, and persons living in high-prevalence areas. In April 2021, the CBTS Pharmacies+ Testing and Surge Testing programs were expanded into the Increasing Community Access to Testing (ICATT) program. As of November 12, 2021, the CBTS and ICATT programs conducted approximately 26.6 million tests with approximately 10,000 active testing sites. Although the CBTS program represented a relatively small portion of overall U.S. SARS-CoV-2 testing, with its successful partnerships and adaptability, the CBTS program serves as a model to guide current community-based screening, surveillance, and disease control programs, and responses to future public health emergencies.

On September 1, 2022, bivalent COVID-19 mRNA vaccines, composed of components from the SARS-CoV-2 ancestral and Omicron BA.4/BA.5 strains, were recommended by the Advisory Committee on Immunization Practices (ACIP) to address reduced effectiveness of COVID-19 monovalent vaccines during SARS-CoV-2 Omicron variant predominance (1). Initial recommendations included persons aged ≥12 years (Pfizer-BioNTech) and ≥18 years (Moderna) who had completed at least a primary series of any Food and Drug Administration-authorized or -approved monovalent vaccine ≥2 months earlier (1). On October 12, 2022, the recommendation was expanded to include children aged 5-11 years. At the time of recommendation, immunogenicity data were available from clinical trials of bivalent vaccines composed of ancestral and Omicron BA.1 strains; however, no clinical efficacy data were available. In this study, effectiveness of the bivalent (Omicron BA.4/BA.5-containing) booster formulation against symptomatic SARS-CoV-2 infection was examined using data from the Increasing Community Access to Testing (ICATT) national SARS-CoV-2 testing program.* During September 14-November 11, 2022, a total of 360,626 nucleic acid amplification tests (NAATs) performed at 9,995 retail pharmacies for adults aged ≥18 years, who reported symptoms consistent with COVID-19 at the time of testing and no immunocompromising conditions, were included in the analysis. Relative vaccine effectiveness (rVE) of a bivalent booster dose compared with that of ≥2 monovalent vaccine doses among persons for whom 2-3 months and ≥8 months had elapsed since last monovalent dose was 30% and 56% among persons aged 18-49 years, 31% and 48% among persons aged 50-64 years, and 28% and 43% among persons aged ≥65 years, respectively. Bivalent mRNA booster doses provide additional protection against symptomatic SARS-CoV-2 in immunocompetent persons who previously received monovalent vaccine only, with relative benefits increasing with time since receipt of the most recent monovalent vaccine dose. Staying up to date with COVID-19 vaccination, including getting a bivalent booster dose when eligible, is critical to maximizing protection against COVID-19 (1).

The recent observations of gravitational waves during the coalescence of black holes has confirmed various predictions of general relativity. However, some concerns have been raised that these putative waves are artifacts. We have analyzed the ten confident gravitational waves detected in Observation Run 1 (O1) and Observation Run 2 (O2) and the gravitational wave emitted by the single known example of a neutron star coalescence, all of which have been verified by the Advanced Laser Interferometer Gravitational Wave Observatory and/or the Advanced Virgo Collaboration. On three occasions both Advanced LIGO and Advanced Virgo detected the same event ( Abbott et al., 2019 ). Whitening of the Hanford data (Gravitational Wave Open Science Center) was employed in this study to reduce colored non-transient noise, but template matching was not employed. In each of the ten events mentioned above we verified the presence of such waves by the application of non-stationary data analysis via the Morlet wavelet. We examined the strain data before, during and after each wave. In each case we employed seven measures of complexity or information before, and during each wave and analyzed these data using doubly multivariate MANOVA, the appropriate statistical test for repeated measures (pre-wave vs wave). In addition we compared the strain data before and after the putative event to see if there was persistence of any effect of the event. In each case the information content was significantly elevated during the wave compared to similar time samples taken before the wave. No persistent effects on the strain data were noted after the event. We conclude our analysis is consistent with the actual detection of gravitational waves by the LIGO and Virgo Collaboration. Our results suggest black hole coalescence generates mathematically complex gravitational waves which may elucidate aspects of such coalescence.

Sustainable aviation fuels (SAFs) have been identified as a method to reduce the aviation sector’s environmental impact through carbon mitigation. Current certification processes of potential, viab...

In this work, a variable-pulse-oscillator is developed and coupled with a burst-mode amplifier for generation of high-energy laser pulses with width of 100 ps to 1 ms and near-Gaussian temporal pulse shape. Pulse energy as high as 600 mJ is demonstrated at 1064 nm, with a super-Gaussian spatial profile and beam quality as good as 1.6 times the diffraction limit. A time-dependent pulse amplification model is developed and is in general agreement with experimentally measured values of output pulse energy and temporal pulse shape of the amplified pulses. Key performance parameters (pulse energy, temporal pulse shape, and spatial beam profile and quality) are analyzed as a function of pulse width across seven orders of magnitude. Additionally, the model is used to elucidate deviations between the simulated and experimental data, showing that the relationship between pulse width and output pulse energy is dominated by the variable-pulse-width oscillator performance, not the burst-mode amplifier.

An optical parametric oscillator (OPO) is developed and characterized for the simultaneous generation of ultraviolet (UV) and near-UV nanosecond laser pulses for the single-shot Rayleigh scattering and planar laser-induced-fluorescence (PLIF) imaging of methylidyne (CH) and nitric oxide (NO) in turbulent flames. The OPO is pumped by a multichannel, 8-pulse Nd:YAG laser cluster that produces up to 225 mJ/pulse at 355 nm with pulse spacing of 100 µs. The pulsed OPO has a conversion efficiency of 9.6% to the signal wavelength of ∼ 430 n m when pumped by the multimode laser. Second harmonic conversion of the signal, with 3.8% efficiency, is used for the electronic excitation of the A-X (1,0) band of NO at ∼ 215 n m , while the residual signal at 430 nm is used for direct excitation of the A-X (0,0) band of the CH radical and elastic Rayleigh scattering. The section of the OPO signal wavelength for simultaneous CH and NO PLIF imaging is performed with consideration of the pulse energy, interference from the reactant and product species, and the fluorescence signal intensity. The excitation wavelengths of 430.7 nm and 215.35 nm are studied in a laminar, premixed C H 4 − H 2 − N H 3 –air flame. Single-shot CH and NO PLIF and Rayleigh scatter imaging is demonstrated in a turbulent C H 4 − H 2 − N H 3 diffusion flame using a high-speed intensified CMOS camera. Analysis of the complementary Rayleigh scattering and CH and NO PLIF enables identification and quantification of the high-temperature flame layers, the combustion product zones, and the fuel-jet core. Considerations for extension to simultaneous, 10-kHz-rate acquisition are discussed.

Layered structures are widely used in optics to control wave reflection and propagation at interfaces but their thickness relative to the wavelength hinders their application in the radio-frequency regime. Here, we design and experimentally demonstrate cascaded metasurfaces with deeply subwavelength thickness that provide both antireflection and wavefront manipulation. We describe the systematic synthesis of the metasurface layers from a prescribed scattering response and demonstrate designs capable of providing near-perfect microwave transmission through glass as well as antireflection focusing of wireless signals from air into water.

Yellow fever virus live attenuated vaccine can rarely cause life-threatening disease. Inherited IFNAR1 deficiency was previously reported in one patient. Here, we report a patient with inherited IFNAR2 deficiency and three other patients with neutralizing auto-antibodies against type I IFNs., Yellow fever virus (YFV) live attenuated vaccine can, in rare cases, cause life-threatening disease, typically in patients with no previous history of severe viral illness. Autosomal recessive (AR) complete IFNAR1 deficiency was reported in one 12-yr-old patient. Here, we studied seven other previously healthy patients aged 13 to 80 yr with unexplained life-threatening YFV vaccine–associated disease. One 13-yr-old patient had AR complete IFNAR2 deficiency. Three other patients vaccinated at the ages of 47, 57, and 64 yr had high titers of circulating auto-Abs against at least 14 of the 17 individual type I IFNs. These antibodies were recently shown to underlie at least 10% of cases of life-threatening COVID-19 pneumonia. The auto-Abs were neutralizing in vitro, blocking the protective effect of IFN-α2 against YFV vaccine strains. AR IFNAR1 or IFNAR2 deficiency and neutralizing auto-Abs against type I IFNs thus accounted for more than half the cases of life-threatening YFV vaccine-associated disease studied here. Previously healthy subjects could be tested for both predispositions before anti-YFV vaccination.

Interactions between turbulent structures (eddies) and premixed flame fronts were characterized with simultaneous CH2O-/OH-PLIF and stereo-PIV at high framing rate (20 kHz). The high temporal resolution of these measurements permitted the tracking and subsequent characterization of the evolution of turbulence–flame interactions. Common events were identified and their effects on the flame structure were assessed, the first of these being that the flame front was locally broadened by large eddies (i.e. those larger than twice the laminar flame thickness). The results suggest that large eddies likely have a two-fold effect on the overall propagation rate, one linked to the increase in burning area (consistent with classical theories) and the other stemming from an enhancement in the local diffusivity experienced by the flame. While this latter point has not been predicted, it has been postulated that large eddies could convect preheated fluid upstream of the flame. To our knowledge the results presented here provide the first conclusive experimental evidence of this phenomenon. The second common event was the persistence of locally thin preheat layers through extremely turbulent flow fields (i.e. those possessing scales much smaller than the laminar flame thickness). Comparisons between sequences of such events and those where broadening occurred indicate that the latter is typically associated with elevated turbulent diffusivity, which supports arguments in favor of using a criterion based on a ratio of diffusivities rather than length scales to predict the onset of preheat-zone broadening.

Understanding of ignition processes is central to design for reliable and safe aerospace combustor systems. Ignition is influenced by many factors including combustor geometry, flow conditions, fuel composition, turbulence intensity, ignition source, and energy deposition method. A toroidal jet-stirred reactor (TJSR) utilizes bulk fluid motion, presence of recirculation zones, a bulk residence time, and turbulence intensities which emulate characteristics relevant to cavity stabilized and swirl stabilized combustors. In this work, a TJSR was used to quantify ignitability and time-to-ignition of premixed ethylene and air. The effects of inlet temperature, residence time, and reactivity were studied on forced ignition processes. Experimental conditions ranged from residence times of 15–35 ms, mixture temperatures of 340–450 K, and equivalence ratios of 0.5–1 using capacitive spark-discharge ignition. The minimum equivalence ratio for ignition (MER), or the equivalence ratio at 50% probability, shows an inverse relationship with mixture temperature and residence time. Prior theory of real engine combustor performance for lean light off, proposed by Ballal and Lefebvre, was compared to the MER and displayed similar trends to the model. Spatially integrated OH* chemiluminescence was used to measure time to ignition within the reactor. Reduction in ignitibility was experienced as the time-to-ignition approached the residence time stressing the importance of device flow time scales in relation to kernel growth dynamics and ignition probability.

Influenza virologic surveillance is critical each season for tracking influenza circulation, following trends in antiviral drug resistance, detecting novel influenza infections in humans, and selecting viruses for use in annual seasonal vaccine production. We developed a framework and process map for characterizing the landscape of US influenza virologic surveillance into 5 tiers of influenza testing: outpatient settings (tier 1), inpatient settings and commercial laboratories (tier 2), state public health laboratories (tier 3), National Influenza Reference Center laboratories (tier 4), and Centers for Disease Control and Prevention laboratories (tier 5). During the 2015–16 season, the numbers of influenza tests directly contributing to virologic surveillance were 804,000 in tiers 1 and 2; 78,000 in tier 3; 2,800 in tier 4; and 3,400 in tier 5. With the release of the 2017 US Pandemic Influenza Plan, the proposed framework will support public health officials in modeling, surveillance, and pandemic planning and response.

Background Detections of influenza A subtype specific antibody responses are often complicated by the presence of cross-reactive antibodies. We developed two novel multiplex platforms for antibody detection. The multiplexed magnetic fluorescence microsphere immunoassay (MAGPIX) is a high throughput laboratory-based assay. Chembio Dual Path Platform (DPP) is a portable and rapid test that could be used in the field. Methods Twelve recombinant globular head domain hemagglutinin (GH HA1) antigens from A(H1N1)pdm09 (pH1N1), A(H2N2), A(H3N2), A(H5N1), A(H7N9), A(H9N2), A(H13N9), B/Victoria lineage, B/Yamagata lineage viruses, and protein A control were used. Human sera from U.S. residents either vaccinated (with H5N1 or pH1N1) or infected with pH1N1 influenza viruses, and sera from live bird market workers in Bangladesh (BDPW) were evaluated. GH HA1 antigens and serum adsorption using full ectodomain recombinant hemagglutinins from A(H1N1) and A(H3N2) were introduced into the platforms to reduce cross-reactivity. Results Serum adsorption reduced cross-reactivity to novel subtype HAs. Compared to traditional hemagglutination inhibition or microneutralization assays, when serum adsorption and the highest fold rise in signals were used to determine positivity, the correct subtype-specific responses were identified in 86% to 100% of U.S. residents exposed to influenza antigens through vaccination or infection (N=49). For detection of H5N1 specific antibodies in sera collected from BDPW, H5 sensitivity was 100% (6/6) for MAGPIX, 83% (5/6) for DPP; H5 specificity was 100% (15/15) and cross-reactivity against other subtype was 0% (0/6) for both platforms. Conclusion MAGPIX and DPP platforms can be utilized for high-throughput and in-field detection of novel influenza virus infections. This article is protected by copyright. All rights reserved.

Quality standards as part of an effective quality management system (QMS) are the cornerstone for generating high-quality test results. Next-generation sequencing (NGS) has the potential to improve both clinical diagnostics and public health surveillance efforts in multiple areas, including infectious diseases. However, the laboratories adopting NGS methods face significant challenges due to the complex and modular process design. This document summarizes the first phase of quality system guidance developed by the Centers for Disease Control and Prevention (CDC) NGS Quality Workgroup. The quality system essentials of personnel, equipment, and process management (quality control and validation) were prioritized based on a risk assessment using information gathered from participating CDC laboratories. Here, we present a prioritized QMS framework, including procedures and documentation tools, to assist laboratory implementation and maintenance of quality practices for NGS workflows.

Ignition processes in scramjet pilot cavities are highly transient events that are influenced by factors including freestream Mach number and inlet geometry, turbulence intensity, cavity geometry, ignition source, and fueling composition and flow rate. In particular, the location of the flame kernel and associated propagation rate of the flame front throughout the cavity can significantly influence the end state of the ignition process. In this work formaldehyde (CH2O) was used as a flame marker to track ignition progress in a plane throughout the span-wise width of the cavity, while chemiluminescence imaging provided path-integrated flame location along the span-wise and axial directions. Planar laser-induced fluorescence (PLIF) excitation utilized the 355 nm frequency-tripled output of an Nd:YAG burst-mode laser operating at 50–100 kHz over 10 ms with available pulse energy up to 80 mJ. Simultaneous CH* chemiluminescence imaging from the top of the cavity was obtained with a high-speed complementary metal-oxide semiconductor camera. A freestream Mach number of 2 with ethylene fuel rates from 55–90 standard liters per minute were examined along with two different ignition sources: a spark discharge and pulse detonator. The resulting formaldehyde PLIF and chemiluminescence images indicate a strong correlation between fueling rate and the delay between the onset of ignition and stable combustion. More importantly, the span-wise propagation rate and structure of the flame front is highly dependent on the fueling rate, burning region, and ignition source.

Planar laser-induced fluorescence (PLIF) of hydroxyl (OH) and formaldehyde (CH

Hybrid femtosecond/picosecond coherent anti-Stokes Raman scattering (fs/ps CARS) of N2 has recently been demonstrated for gas-phase thermometry in reacting flows, enabling frequency-domain detection at high repetition rates with excellent chemical specificity and independence from the effects of collisions and nonresonant background. In this work, we overcome the limited sensitivity of vibrational fs/ps CARS thermometry of N2 below 1200 K by spectrally resolving J-dependent rovibrational coherence revivals that occur 32 ps after initial excitation. The N2 rovibrational coherence is excited using broadband, 100-fs pump and Stokes pulses and probed as a function of time using a narrowband, 5.8-ps probe pulse (bandwidth of 2.5 cm−1). The rovibrational features exhibit sufficient temperature sensitivity below 1200 K for accurate thermometry using a simple, time-dependent phenomenological model. Specifically, three distinct spectral features at a single probe delay of 32.5 ps are analyzed, corresponding to rovibrational revivals with mean rotational quantum numbers of = 3.5, 14, and 28. Good agreement is found between simulated and measured fs/ps CARS spectra in an adiabatic flat-flame burner from 298 to 2400 K. Copyright © 2015 John Wiley & Sons, Ltd.

Objective The aim of this study is to describe the clinical phenotype of alcohol use disorder (AUD) treatment-seeking patients with Roux-en-Y Gastric Bypass Surgery (RYGB) history; and to compare it to AUD obese non-RYGB controls. Methods Retrospective study of electronic medical records for all patients 30–60 years treated at the Mayo Clinic Addiction Treatment Program, between June, 2004 and July, 2012. Comparisons were performed with consumption patterns pre-RYGB and at time of treatment; excluding patients with AUD treatments pre-RYGB. Results Forty-one out of 823 patients had a RYGB history (4.9%); 122 controls were selected. Compared to controls, the RYGB group had significantly more females [ n = 29 (70.7%) vs. n = 35 (28.7%) p p = 0.002). On average, RYGB patients reported resuming alcohol consumption 1.4 ± 0.2 years post-surgery, meeting criteria for AUD at 3.1 ± 0.5 years and seeking treatment at 5.4 ± 0.3 years postoperatively. Pre-surgical drinks per day were significantly fewer compared to post-surgical consumption [2.5 ± 0.4 vs. 8.1 ± 1.3, p = 0.009]. Prior to admission, RYGB patients reported fewer drinking days per week vs. controls (4.7 ± 0.3 vs. 5.5 ± 1.8 days, p = 0.02). Neither RYGB, gender, age nor BMI was associated with differential drinking patterns. Conclusion The results of this study suggest that some patients develop progressive AUD several years following RYGB. This observation has important clinical implications, calling for AUD-preventive measures following RYGB. Further large-scale longitudinal studies are needed to clarify the association between RYGB and AUD onset.

CD8 T cells are a potent tool for eliminating intracellular pathogens and tumor cells. Thus, eliciting robust CD8 T-cell immunity is the basis for many vaccines under development. However, the relationship between antigen load and the magnitude of the CD8 T-cell response is not well-described in a human immune response. Here we address this issue by quantifying viral load and the CD8 T-cell response in a cohort of 80 individuals immunized with the live attenuated yellow fever vaccine (YFV-17D) by sampling peripheral blood at days 0, 1, 2, 3, 5, 7, 9, 11, 14, 30, and 90. When the virus load was below a threshold (peak virus load < 225 genomes per mL, or integrated virus load < 400 genome days per mL), the magnitude of the CD8 T-cell response correlated strongly with the virus load (R(2) ∼ 0.63). As the virus load increased above this threshold, the magnitude of the CD8 T-cell responses saturated. Recent advances in CD8 T-cell-based vaccines have focused on replication-incompetent or single-cycle vectors. However, these approaches deliver relatively limited amounts of antigen after immunization. Our results highlight the requirement that T-cell-based vaccines should deliver sufficient antigen during the initial period of the immune response to elicit a large number of CD8 T cells that may be needed for protection.

Treatment of chemical dependence ("addiction") requires an understanding of its effects on the brain. To guide research in the area of chemical dependence, several foundational theories have been developed. These include the incentive salience, receptor down-regulation, opponent process, and psychomotor stimulant theories. These have been important both in summarizing and in guiding investigations. However, the extant theories do not provide a single unified framework nor have they yielded all of the guidance necessary for effective chemical dependence treatment. The present paper summarizes and then integrates these theories and suggests some implications for the treatment followed by this integration.

Simultaneous high-speed quantitative imaging of mixture fraction and velocity is demonstrated using the fourth- and second-harmonic outputs, respectively, of a burst-mode Nd:YAG laser. A tenfold increase in the record length and 16-fold increase in per-pulse energy are achieved compared with previous measurements of mixture fraction using burst-mode and continuously pulsed diode-pumped solid-state lasers, respectively. The high output energy is used for quantitative, high-speed mixture-fraction imaging with acetone planar laser-induced fluorescence, which also enables simultaneous particle-based velocimetry without interference from particle scattering. A semiquantitative model is used to determine the limitations on fourth-harmonic output energy due to the effects of transient absorption and thermal phase mismatch over a range of repetition rates. Data are presented for mixing within a turbulent jet (Reynolds number of 15,000) and are validated by comparisons with known turbulent mixing laws and previously published data.

See-through-wall coherent microwave scattering from resonance-enhanced multiphoton ionization (REMPI) for rotational temperature measurements of molecular oxygen has been developed and demonstrated in a flow reactor at atmospheric pressure. Through limited, single-ended optical access, a laser beam was focused to generate local ionization of molecular oxygen in a heated quartz flow reactor enclosed by ceramic heating elements. Coherent microwaves were transmitted, and the subsequent scattering off the laser-induced plasma was received, through the optically opaque ceramic heater walls and used to acquire rotational spectra of molecular oxygen and to determine temperature. Both axial and radial air-temperature profiles were obtained in the flow reactor with an accuracy of ±20 K⁢(±5%). The experimental results show good agreement with a steady-state computational heat transfer model. This technique shows great potential for non-invasive, high-fidelity measurement of spatially localized temperature and radical species concentration in combustion kinetic experiments and confined combustors constructed of advanced ceramic materials in which limited or non-existing optical access hinders usage of conventional optical diagnostic techniques to quantify thermal non-uniformity.

Hybrid femtosecond/picosecond (fs/ps) vibrational coherent anti-Stokes Raman scattering was demonstrated for high-speed, in-situ measurements of CH4/N2 mole fractions in binary gas mixtures with temporal nonresonant suppression. A single broadband, 100-fs laser source was used to simultaneously excite CH4 and N2 vibrational Raman transitions, characterized by a spectral separation of ~584 cm−1, followed by frequency-domain detection with a time-delayed ps probe pulse. In this manner, the temporal evolution of the nonresonant background and vibrational wavepackets of CH4 and N2 could be evaluated to maximize signal and minimize interferences. A typical accuracy of ~95% was achieved for experimental CH4 mole fractions between 0.02 and 0.70, and a precision of ~4% was achieved by maximizing relative excitation energy through induced chirp in the pump and Stokes pulses. It was further shown that mole-fraction measurements were not affected by variations in bi-molecular broadened linewidths over a wide range of mixture fractions. Copyright © 2013 John Wiley & Sons, Ltd.

100-kHz particle image velocimetry (PIV) is demonstrated using a double-pulsed, burst-mode laser with a burst duration up to 100 ms. This enables up to 10,000 time-sequential vector fields for capturing a temporal dynamic range spanning over three orders of magnitude in high-speed turbulent flows. Pulse doublets with inter-pulse spacing of 2 µs and repetition rate of 100 kHz are generated using a fiber-based oscillator and amplified through an all-diode-pumped, burst-mode amplifier. A physics-based model of pulse doublet amplification in the burst-mode amplifier is developed and used to accurately predict oscillator pulse width and pulse intensity inputs required to generate equal-energy pulse doublets at 532 nm for velocity measurements. The effect of PIV particle response and high-speed-detector limitations on the spatial and temporal resolution are estimated in subsonic turbulent jets. An effective spatial resolution of 266–275 µm and temporal resolution of 10 µs are estimated from the 8 × 8 pixel correlation window and inter-doublet time spacing, respectively. This spatiotemporal resolution is sufficient for quantitative assessment of integral time and length scales in highly turbulent jets with Reynolds numbers in the range 15,000–50,000. The temporal dynamic range of the burst-mode PIV measurement is 1200, limited by the 85-ms high-energy portion of the burst and 30-kHz high-frequency noise limit.

Two-dimensional gas-phase coherent anti-Stokes Raman scattering (2D-CARS) thermometry is demonstrated at 1 kHz in a heated jet. A hybrid femtosecond/picosecond CARS configuration is used in a two-beam phase-matching arrangement with a 100-femtosecond pump/Stokes pulse and a 107-picosecond probe pulse. The femtosecond pulse is generated using a mode-locked oscillator and regenerative amplifier that is synchronized to a separate picosecond oscillator and burst-mode amplifier. The CARS signal is spectrally dispersed in a custom imaging spectrometer and detected using a high-speed camera with image intensifier. 1-kHz, single-shot planar measurements at room temperature exhibit error of 2.6% and shot-to-shot variations of 2.6%. The spatial variation in measured temperature is 9.4%. 2D-CARS temperature measurements are demonstrated in a heated Osub2/subjet to capture the spatiotemporal evolution of the temperature field.

Influenza hemagglutination inhibition (HI) and virus microneutralization assays (MN) are widely used for seroprevalence studies. However, these assays have limited field portability and are difficult to fully automate for high throughput laboratory testing. To address these issues, three multiplex influenza subtype-specific antibody detection assays were developed using recombinant hemagglutinin antigens in combination with Chembio, Luminex®, and ForteBio® platforms. Assay sensitivity, specificity, and subtype cross-reactivity were evaluated using a panel of well characterized human sera. Compared to the traditional HI, assay sensitivity ranged from 87% to 92% and assay specificity in sera collected from unexposed persons ranged from 65% to 100% across the platforms. High assay specificity (86–100%) for A(H5N1) rHA was achieved for sera from exposed or unexposed to hetorosubtype influenza HAs. In contrast, assay specificity for A(H1N1)pdm09 rHA using sera collected from A/Vietnam/1204/2004 (H5N1) vaccinees in 2008 was low (22–30%) in all platforms. Although cross-reactivity against rHA subtype proteins was observed in each assay platform, the correct subtype specific responses were identified 78% to 94% of the time when paired samples were available for analysis. These results show that high throughput and portable multiplex assays that incorporate rHA can be used to identify influenza subtype specific infections.

Nitric oxide planar laser-induced fluorescence (NO PLIF) imaging is demonstrated at a 10-kHz repetition rate in the Calspan University at Buffalo Research Center’s (CUBRC) 48-inch Mach 9 hypervelocity shock tunnel using a pulse burst laser–based high frame rate imaging system. Sequences of up to ten images are obtained internal to a supersonic combustor model, located within the shock tunnel, during a single ~10-millisecond duration run of the ground test facility. Comparison with a CFD simulation shows good overall qualitative agreement in the jet penetration and spreading observed with an average of forty individual PLIF images obtained during several facility runs.

Fully mature enamel is about 98% mineral by weight. While mineral crystals appear very early during its formative phase, the newly secreted enamel is a soft gel-like matrix containing several enamel matrix proteins of which the most abundant is amelogenin ( Amelx). Histological analysis of mineralized dental enamel reveals markings called cross-striations associated with daily increments of enamel formation, as evidenced by injections of labeling dyes at known time intervals. The daily incremental growth of enamel has led to the hypothesis that the circadian clock might be involved in the regulation of enamel development. To identify daily rhythms of clock genes and Amelx, we subjected murine ameloblast cells to serum synchronization to analyze the expression of the circadian transcription factors Per2 and Bmal1 by real-time PCR. Results indicate that these key genetic regulators of the circadian clock are expressed in synchronized murine ameloblast cell cultures and that their expression profile follows a circadian pattern with acrophase and bathyphase for both gene transcripts in antiphase. Immunohistological analysis confirms the protein expression of Bmal and Cry in enamel cells. Amelx expression in 2-day postnatal mouse molars dissected every 4 hours for a duration of 48 hours oscillated with an approximately 24-hour period, with a significant approximately 2-fold decrease in expression during the dark period compared to the light period. The expression of genes involved in bicarbonate production ( Car2) and transport ( Slc4a4), as well as in enamel matrix endocytosis ( Lamp1), was greater during the dark period, indicating that ameloblasts express these proteins when Amelx expression is at the nadir. The human and mouse Amelx genes each contain a single nonconserved E-box element within 10 kb upstream of their respective transcription start sites. We also found that within 2 kb of the transcription start site of the human NFYA gene, which encodes a positive regulator of amelogenin, there is an E-box element that is conserved in rodents and other mammals. Moreover, we found that Nfya expression in serum-synchronized murine ameloblasts oscillated with a strong 24-hour rhythm. Taken together, our data support the hypothesis that the circadian clock temporally regulates enamel development.

The only extraterrestrial life detection experiments ever conducted were the three which were components of the 1976 Viking Mission to Mars. Of these, only the Labeled Release experiment obtained a clearly positive response. In this experiment 14 C radiolabeled nutrient was added to the Mars soil samples. Active soils exhibited rapid, substantial gas release. The gas was probably CO2 and, possibly, other radiocarbon-containing gases. We have applied complexity analysis to the Viking LR data. Measures of mathematical complexity permit deep analysis of data structure along continua including signal vs. noise, entropy vs.negentropy, periodicity vs. aperiodicity, order vs. disorder etc. We have employed seven complexity variables, all derived from LR data, to show that Viking LR active responses can be distinguished from controls via cluster analysis and other multivariate techniques. Furthermore, Martian LR active response data cluster with known biological time series while the control data cluster with purely physical measures. We conclude that the complexity pattern seen in active experiments strongly suggests biology while the different pattern in the control responses is more likely to be non-biological. Control responses that exhibit relatively low initial order rapidly devolve into near-random noise, while the active experiments exhibit higher initial order which decays only slowly. This suggests a robust biological response. These analyses support the interpretation that the Viking LR experiment did detect extant microbial life on Mars.

Real-time PCR methodology can be applied to rapidly and accurately detect influenza viruses. During times of surge testing or enhanced pandemic surveillance, public health laboratories (PHLs) may experience overwhelming demand for testing, even while the prevalence of positive specimens remains low. To improve laboratory capacity and testing efficiency during surges, we evaluated whether nasopharyngeal (NP)/throat swab specimens can be pooled and tested for the presence of the 2009 H1N1 influenza virus without a reduction in sensitivity. Pools of 10 specimens were extracted and concentrated upon elution on the MagNA Pure LC instrument, and real-time PCR was performed on the Applied Biosystems 7500 Fast platform, using the CDC swine influenza virus real-time RT-PCR detection panel (rRT-PCR swine flu panel). Specimens in positive pools were singly re-extracted and retested by PCR to identify individual positive samples. Initial studies showed that spiking a pool of nine negative specimens (100 μl each) or 900 μl of virus transport medium with 100 μl of a positive clinical specimen caused no loss of sensitivity by rRT-PCR testing. Pools containing either multiple positive specimens or specimens positive for other respiratory viruses also showed no negative effect on crossing threshold ( C T ) values. To test the robustness of the pooling protocol, a panel of 50 blinded samples was sent to three PHLs and tested in five pools of 10. All PHLs correctly identified the positive specimens. This study demonstrates the feasibility of using a pooling strategy to increase capacity and conserve resources during surge testing and periods of enhanced influenza surveillance when the prevalence is low.

These tools enabled laboratory response to the 10-fold increase in testing demands., A novel influenza A (H1N1) virus detected in April 2009 rapidly spread around the world. North American provincial and state laboratories have well-defined roles and responsibilities, including providing accurate, timely test results for patients and information for regional public health and other decision makers. We used the multidisciplinary response and rapid implementation of process changes based on Lean methods at the provincial public health laboratory in British Columbia, Canada, to improve laboratory surge capacity in the 2009 influenza pandemic. Observed and computer simulating evaluation results from rapid processes changes showed that use of Lean tools successfully expanded surge capacity, which enabled response to the 10-fold increase in testing demands.

T cell dysfunction is an important feature of many chronic viral infections. In particular, it was shown that programmed death-1 (PD-1) regulates T cell dysfunction during chronic lymphocytic choriomeningitis virus infection in mice, and PD-1hi cells exhibit an intense exhausted gene signature. These findings were extended to human chronic infections such as HIV, hepatitis C virus, and hepatitis B virus. However, it is not known if PD-1hi cells of healthy humans have the traits of exhausted cells. In this study, we provide a comprehensive description of phenotype, function, and gene expression profiles of PD-1hi versus PD-1lo CD8 T cells in the peripheral blood of healthy human adults as follows: 1) the percentage of naive and memory CD8 T cells varied widely in the peripheral blood cells of healthy humans, and PD-1 was expressed by the memory CD8 T cells; 2) PD-1hi CD8 T cells in healthy humans did not significantly correlate with the PD-1hi exhausted gene signature of HIV-specific human CD8 T cells or chronic lymphocytic choriomeningitis virus-specific CD8 T cells from mice; 3) PD-1 expression did not directly affect the ability of CD8 T cells to secrete cytokines in healthy adults; 4) PD-1 was expressed by the effector memory compared with terminally differentiated effector CD8 T cells; and 5) finally, an interesting inverse relationship between CD45RA and PD-1 expression was observed. In conclusion, our study shows that most PD-1hi CD8 T cells in healthy adult humans are effector memory cells rather than exhausted cells.