A STING-dependent innate-sensing pathway mediates resistance to corneal HSV-1 infection via upregulation of the antiviral effector tetherin.

Type 1 interferons (IFNs; IFNα/ß) mediate immunological host resistance to numerous viral infections, including herpes simplex virus type 1 (HSV-1). The pathways responsible for IFNα/ß signaling during the innate immune response to acute HSV-1 infection in the cornea are incompletely understood. Using a murine ocular infection model, we hypothesized that the stimulator of IFN genes (STING) mediates resistance to HSV-1 infection at the ocular surface and preserves the structural integrity of this mucosal site. Viral pathogenesis, tissue pathology, and host immune responses during ocular HSV-1 infection were characterized by plaque assay, esthesiometry, pachymetry, immunohistochemistry, flow cytometry, and small interfering RNA transfection in wild-type C57BL/6 (WT), STING-deficient (STING(-/-)), and IFNα/ß receptor-deficient (CD118(-/-)) mice at days 3-5 postinfection. The presence of STING was critical for sustained control of HSV-1 replication in the corneal epithelium and resistance to viral neuroinvasion, but loss of STING had a negligible impact with respect to gross tissue pathology. Auxiliary STING-independent IFNα/ß signaling pathways were responsible for maintenance of corneal integrity. Lymphatic vessels, mast cells, and sensory innervation were compromised in CD118(-/-) mice concurrent with increased tissue edema. STING-dependent signaling led to the upregulation of tetherin, a viral restriction factor we identify is important in containing the spread of HSV-1 in vivo.

The Current State of Vaccine Development for Ocular HSV-1 Infection.

HSV-1 continues to be the leading cause of infectious corneal blindness. Clinical trials for vaccines against genital HSV infection have been ongoing for more than three decades. Despite this, no approved vaccine exists, and no formal clinical trials have evaluated the impact of HSV vaccines on eye health. We review here the current state of development for an efficacious HSV-1 vaccine and call for involvement of ophthalmologists and vision researchers.

Characterization of mechanical properties of a hollow cylinder with zero group velocity Lamb modes.

Hollow cylinders used in the industry must be regularly inspected. Elastic guided waves, similar to Lamb modes in a plate, can propagate in the axial direction or around the circumference. They are sensitive to geometrical and mechanical parameters of the cylindrical shell. The objective of this paper is to show that zero group velocity (ZGV) Lamb modes can be used to bring out anisotropy and to measure elastic constants of the material. This study provides experimental and numerical investigations on a Zirconium alloy tube extensively used by the nuclear industry in reactor core components. A non-contact method, based on laser ultrasound techniques and ZGV Lamb modes, demonstrates that the difference observed between axial and circumferential guided waves cannot be explained by an isotropic model. Then, a transverse isotropic model is used for the Zircaloy tube. Four of the five elastic constants are directly extracted from ZGV resonance frequencies. The last one is deduced from the measured dispersion spectra. With this complete set of constants, a good agreement is obtained between theoretical and experimental dispersion curves for both axially and circumferentially propagating guided waves.

Geobiological constraints on Earth system sensitivity to CO2 during the Cretaceous and Cenozoic.

Earth system climate sensitivity (ESS) is the long-term (>10³ year) response of global surface temperature to doubled CO2 that integrates fast and slow climate feedbacks. ESS has energy policy implications because global temperatures are not expected to decline appreciably for at least 10³ year, even if anthropogenic greenhouse gas emissions drop to zero. We report provisional ESS estimates of 3 °C or higher for some of the Cretaceous and Cenozoic based on paleo-reconstructions of CO2 and temperature. These estimates are generally higher than climate sensitivities simulated from global climate models for the same ancient periods (approximately 3 °C). Climate models probably do not capture the full suite of positive climate feedbacks that amplify global temperatures during some globally warm periods, as well as other characteristic features of warm climates such as low meridional temperature gradients. These absent feedbacks may be related to clouds, trace greenhouse gases (GHGs), seasonal snow cover, and/or vegetation, especially in polar regions. Better characterization and quantification of these feedbacks is a priority given the current accumulation of atmospheric GHGs.

Edge resonance and zero group velocity Lamb modes in a free elastic plate.

The local resonances of a free isotropic elastic plate are investigated using laser ultrasonic techniques. Experimental results are interpreted in terms of zero group velocity Lamb modes and edge mode. At a distance from the edge larger than the plate thickness a sharp resonance is observed at the frequency where the group velocity of the first symmetrical Lamb mode vanishes. Close to the edge of the plate, the resonance due to the edge mode dominates. Both zero group velocity and edge resonances appear at the theoretically predicted frequencies. These frequencies do not vary with the distance from the edge of the plate and the transition between the two modes of vibration, at about the plate thickness, is abrupt. Using a laser excitation on the edge, the amplitude profile of the normal displacement at the edge resonance frequency was determined.

Numerical predictions and experiments for optimizing hidden corrosion detection in aircraft structures using Lamb modes.

To increase the sensitivity of Lamb waves to hidden corrosion in aircraft structures, a preliminary step is to understand the phenomena governing this interaction. A hybrid model combining a finite element approach and a modal decomposition method is used to investigate the interaction of Lamb modes with corrosion pits. The finite element mesh is used to describe the region surrounding the corrosion pits while the modal decomposition method permits to determine the waves reflected and transmitted by the damaged area. Simulations make easier the interpretation of some parts of the measured waveform corresponding to superposition of waves diffracted by the corroded area. Numerical results permit to extract significant information from the transmitted waveform and thus to optimize the signal processing for the detection of corrosion at an early stage. Now, we are able to detect corrosion pits down to 80-mum depth distributed randomly on a square centimeter of an aluminum plate. Moreover, thickness variations present on aircraft structures can be discriminated from a slightly corroded area. Finally, using this experimental setup, aircraft structures have been tested.

A combined finite element and modal decomposition method to study the interaction of Lamb modes with micro-defects.

This paper presents a combined finite element and modal decomposition method to study the interaction of Lamb waves with damaged area. The finite element mesh is used to describe the region around the defects. On the contrary to other hybrid models already developed, the interaction between Lamb waves and defects is computed in the temporal domain. Then, the modal decomposition method permits to determine the wave reflected and transmitted by the damaged area. Modal analysis allows also identifying the mode conversions induced by the defects. These numerical results agree with previous finite element results concerning the interaction of Lamb modes with a notch. Experiments, carried out with gauged defects on an aluminum plate, are also compared to numerical predictions to validate the simulation. Compared to classical techniques of simulation, this new method allows us to investigate the interaction of Lamb modes generated at high frequency-thickness product with micro-defects as corrosion pitting.

Quantitative analysis of Fusarium mycotoxins in maize using accelerated solvent extraction before liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry.

A method for the simultaneous quantitative determination of deoxynivalenol (DON), fumonisin B1 (FB1) and zearalenone (ZEN) in maize by liquid chromatography-atmospheric pressure chemical ionization tandem mass spectrometry (LC-APCIMS/MS), using stable isotopically labelled and structural analogues internal standards, is described. The procedure involves accelerated solvent extraction followed by two solid-phase clean-up steps on strong anion exchange resin and a Mycosep column. Typical recoveries were calculated by spiking blank maize at three different concentrations for deoxynivalenol (200, 400 and 1000 microg kg(-1)) at 70%, for fumonisin B1 (100, 200 and 1000 microg kg(-1)) at 90%, and for zearalenone (50, 100 and 200 microg kg(-1)) at 40%. LC-APCIMS/MS analyses were realized in collision-induced dissociation on an ion-trap instrument to provide a high degree of selectivity and sensitivity. Extraction of ions from two transition reactions, monitored by LC-APCIMS/MS for each analyte, enabled a limit of detection for DON, FB1 and ZEN at, respectively, 10, 20 and 3 microg kg(-1), and a limit of quantification at, respectively, 50, 50 and 10 microg kg(-1). The robustness of the method was also evaluated with the analysis of wheat samples.

Analysis of chloramphenicol in honeys of different geographical origin by liquid chromatography coupled to electrospray ionization tandem mass spectrometry.

A sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed to detect trace amounts of the antibiotic chloramphenicol (CAP) in honey. The methodology entailed a solid-phase extraction of aqueous honey solutions followed by liquid-liquid partitioning, filtration and direct injection onto the LC-MS/MS system. Honey extracts were spiked with an isotopically labelled internal standard (d(5)-CAP) to compensate for analyte loss and potential ion suppression during the MS stage. Detection of the analyte was achieved by negative ionization electrospray in the selected reaction monitoring (SAM) mode. For confirmation, four characteristic mass transitions were monitored each for the analyte and the surrogate standard. The method was validated according to the latest European Union criteria for the analyses of veterinary drug residues in food. At all three fortification levels studied (0.1, 0.2, 0.5 microg kg(-1)) the method was accurate to within 15%. The repeatability and within-laboratory reproducibilities were <12 and 18%, respectively. The decision limit (CC alpha) and detection capability (CC beta) were both <0.1 microg kg(-1). The procedure provides a sensitive and reliable method for the determination of residues of chloramphenicol in honey. Numerous raw honeys of various geographical origins were analysed, showing extensive contamination particularly those of Chinese origin.

Detection of cracks in a thin air-filled hollow cylinder by application of the DORT method to elastic components of the echo.

The DORT method (Decomposition de l'Opérateur de Retournement Temporel in French) is a scattering analysis technique which uses arrays of transducers. This method is efficient for detection of selective focusing on point-like scatterers. It has been also applied to analyze the scattering by an air-filled cylindrical steel shell immersed in water. It was shown that the diagonalization of the time reversal operator allows us to separate the different elastic components of the scattered field. Here, we apply the method to detect flaws in hollow cylinders. In this case, the dominant components are the three circumferential waves (A0, A1 and S0 Lamb modes). Each Lamb mode corresponds to an invariant of the time reversal operator. The dispersion curves of these waves are calculated from these invariants. Resonance frequencies of the shell are deduced from the frequency dependence of the eigenvalues of the time reversal operator. It is shown that the presence of a crack (0.2 mm in depth) affects significantly the eigenvalue distribution of the time reversal operator. Thus, the DORT method offers a new means for detecting defects in a shell.

Nondestructive evaluation of cylindrical parts using laser ultrasonics.

We applied the laser ultrasonic technique for detecting surface breaking slots in steel cylinders (25 mm in diameter). The observation of the detected signal over a long time (500 micros), shows that the interaction of the two contra-propagating incident Rayleigh waves reinforce the echoes coming from the defect. These echoes are slowly growing with time whereas the main signals decrease. This energy transfer occurring at each revolution of the waves around the cylinder allows the detection of cracks having a depth (h approximately 80 microm), very small compared to the Rayleigh wavelength (lambda approximately 2 mm). The evaluation of the material was performed by processing the detected signal in a sliding time window. A cross-correlation is made either between a reference signal and the signal from the tested sample or between two signals probed for two different positions of the sample. In both cases, the slope of the cross-correlation coefficient versus the number of turns is proportional to the depth of the slot.

An atmospheric pCO2 reconstruction across the Cretaceous-Tertiary boundary from leaf megafossils.

The end-Cretaceous mass extinctions, 65 million years ago, profoundly influenced the course of biotic evolution. These extinctions coincided with a major extraterrestrial impact event and massive volcanism in India. Determining the relative importance of each event as a driver of environmental and biotic change across the Cretaceous-Tertiary boundary (KTB) crucially depends on constraining the mass of CO(2) injected into the atmospheric carbon reservoir. Using the inverse relationship between atmospheric CO(2) and the stomatal index of land plant leaves, we reconstruct Late Cretaceous-Early Tertiary atmospheric CO(2) concentration (pCO(2)) levels with special emphasis on providing a pCO(2) estimate directly above the KTB. Our record shows stable Late Cretaceous/Early Tertiary background pCO(2) levels of 350-500 ppm by volume, but with a marked increase to at least 2,300 ppm by volume within 10,000 years of the KTB. Numerical simulations with a global biogeochemical carbon cycle model indicate that CO(2) outgassing during the eruption of the Deccan Trap basalts fails to fully account for the inferred pCO(2) increase. Instead, we calculate that the postboundary pCO(2) rise is most consistent with the instantaneous transfer of approximately 4,600 Gt C from the lithic to the atmospheric reservoir by a large extraterrestrial bolide impact. A resultant climatic forcing of +12 W.m(-2) would have been sufficient to warm the Earth's surface by approximately 7.5 degrees C, in the absence of counter forcing by sulfate aerosols. This finding reinforces previous evidence for major climatic warming after the KTB impact and implies that severe and abrupt global warming during the earliest Paleocene was an important factor in biotic extinction at the KTB.

Reading a CO2 signal from fossil stomata.

The inverse relationship between atmospheric CO2 and the stomatal index (proportion of epidermal cells that are stomata) of vascular land plant leaves has led to the use of fossil plant cuticles for determining ancient levels of CO2 . In contemporary plants the stomatal index repeatedly shows a lower sensitivity atmospheric CO2 levels above 340 ppm in the short term. These observations demonstrate that the phenotypic response is nonlinear and may place constraints on estimating higher-than-present palaeo-CO2 levels in this way. We review a range of evidence to investigate the nature of this nonlinearity. Our new data, from fossil Ginkgo cuticles, suggest that the genotypic response of fossil Ginkgo closely tracks the phenotypic response seen in CO2 enrichment experiments. Reconstructed atmospheric CO2 values from fossil Ginkgo cuticles compare well with the stomatal ratio method of obtaining a quantitative CO2 signal from extinct fossil plants, and independent geochemical modelling studies of the long-term carbon cycle. Although there is self-consistency between palaeobiological and geochemical CO2 estimates, it should be recognized that the nonlinear response is a limitation of the stomatal approach to estimating high palaeo-CO2 levels.

Stomatal density and stomatal index as indicators of paleoatmospheric CO(2) concentration.

A growing number of studies use the plant species-specific inverse relationship between atmospheric CO(2) concentration and stomatal density (SD) or stomatal index (SI) as a proxy for paleo-CO(2) levels. A total of 285 previously published SD and 145 SI responses to variable CO(2) concentrations from a pool of 176 C(3) plant species are analyzed here to test the reliability of this method. The percentage of responses inversely responding to CO(2) rises from 40 and 36% (for SD and SI, respectively) in experimental studies to 88 and 94% (for SD and SI, respectively) in fossil studies. The inconsistent experimental responses verify previous concerns involving this method, however the high percentage of fossil responses showing an inverse relationship clearly validates the method when applied over time scales of similar length. Furthermore, for all groups of observations, a positive relationship between CO(2) and SD/SI is found in only

Paleobotanical evidence for near present-day levels of atmospheric Co2 during part of the tertiary.

Understanding the link between the greenhouse gas carbon dioxide (CO(2)) and Earth's temperature underpins much of paleoclimatology and our predictions of future global warming. Here, we use the inverse relationship between leaf stomatal indices and the partial pressure of CO(2) in modern Ginkgo biloba and Metasequoia glyptostroboides to develop a CO(2) reconstruction based on fossil Ginkgo and Metasequoia cuticles for the middle Paleocene to early Eocene and middle Miocene. Our reconstruction indicates that CO(2) remained between 300 and 450 parts per million by volume for these intervals with the exception of a single high estimate near the Paleocene/Eocene boundary. These results suggest that factors in addition to CO(2) are required to explain these past intervals of global warmth.

A study of the secular equation for rayleigh waves using the root locus method.

In a recent note Rahman and Barber [Trans. ASME 62 (1995) 250] proposed an exact expression for the roots of the secular equation for the Rayleigh waves in an isotropic half-space. Using the root locus method, we describe in a very simpler manner the evolution of the roots versus the Poisson's ratio and we derive easily the critical value for which the nature of the roots changes.

Diffraction effects in the parametric interaction of acoustic waves: application to measurements of the nonlinearity parameter B/A in liquids.

This paper deals with diffraction effects related to the determination of nonlinearity parameters B/A in liquids using the parametric interaction. A diffraction model, based on plane wave expansions, is applied to weak nonlinear interactions between two ultrasonic waves propagating in an absorbing medium. A validation of the model has been carried out with an experiment performed using an optical interferometer. A method is proposed to measure nonlinearity parameters with the parametric interaction of two waves with a high frequency ratio. With respect to diffraction effects, the parametric interaction can then be identified with a phase modulation of the high frequency wave. A prototype, using the interaction of a 30-MHz frequency continuous wave with an acoustic pulse of central frequency 2.5 MHz, has been designed. The proposed method is validated by nonlinearity parameters measurements in well-known liquids (water and ethanol). The uncertainty on absolute measurements is discussed, and a relative method is proposed to increase the accuracy. This method enables measurements without any transducer calibration.

A preliminary study of sonographic grading of fetal intracardiac echogenic foci: feasibility, reliability and association with aneuploidy.

OBJECTIVES: To prospectively and quantitatively grade intracardiac echogenic focus/foci (ICEF) using sonographic gain reduction and to determine the association of ICEF by grade with fetal aneuploidy. METHODS: Women referred for raised maternal age (> or = 35 years), or > 18 years of age and with a Down syndrome risk > or = 1/270, increased trisomy 18 risk by second trimester serum screen or a prior aneuploid offspring were included in this institutionally approved protocol. Only pregnancies of gestational age between 14 and 24 weeks were included. All women had a targeted ultrasound and were offered fetal chromosome analysis. The classification of ICEF was made from a four-chamber view of the fetal heart. The echo amplitude of the ICEF was compared to that of the thoracic spine and categorized according to the comparative gain setting at which the image of the relevant structure disappeared: Grade O = no ICEF present, Grade 1 = ICEF image lost before thoracic spine when gain was reduced, Grade 2 = ICEF image lost at same gain setting as thoracic spine, Grade 3 = thoracic spine image lost before ICEF. The primary outcome was a prenatally or post-natally detected chromosomal abnormality. RESULTS: A total of 885 eligible women were examined during the 21-month study period. ICEF were seen in 29 (3.3%) fetuses: 24(83%) in the left ventricle and five (17%) in the right ventricle. A chromosome abnormality was identified in 13/671 (1.9%) fetuses without ICEF (Grade 0) and 0/21 (0%) fetuses with Grade 1 ICEF. In contrast, two of five (40%) fetuses with Grade 2 ICEF were aneuploid (P = 0.005). No Grade 3 ICEF were observed. Additional sonographic abnormalities were seen in both aneuploid fetuses with Grade 2 ICEF. Interobserver agreement on ICEF grading was noted in 50/50 (100%) examinations (kappa = 1.0). CONCLUSIONS: Sonographic grading of ICEF is feasible and highly reliable. Grade 2 ICEF, especially when accompanied by additional sonographic markers of a chromosomal abnormality, are associated with aneuploidy significantly more frequently than Grade 1 ICEF.

Efficacy of community-based second trimester genetic ultrasonography in detecting the chromosomally abnormal fetus.

We sought to assess prospectively the efficacy of community-based genetic ultrasonography in detecting chromosomally abnormal fetuses in a high-risk population and determine independent markers of aneuploidy. Patients 18 years old and older who were between 14 and 24 weeks' gestation were included if referred for maternal age greater than 35 years, increased risk of Down syndrome or trisomy 18 by second trimester serum screen, or prior affected offspring. All women had a targeted ultrasonographic examination between April 1997 and June 1999 and were offered fetal chromosomal analysis. Markers of aneuploidy and pregnancy outcomes were recorded prospectively. The primary outcome was prenatally or postnatally detected chromosomal abnormalities. Of the 1030 fetuses seen during the study, 789 had outcome data available and constituted the study group. In this group, 694 (87.9%) ultrasonograms were normal, 73 (9.2%) had one marker present, 17 (2.2%) had two markers present, and 5 (0.6%) had three or more markers present. Fourteen of 17 (82.3%) aneuploid fetuses had an abnormal ultrasonogram (one or more markers present), including 5 of 7 (71.4%) with Down syndrome. Logistic regression showed abnormal four-chamber view, structural anomaly, and intracardiac echogenic focus to be significant aneuploidy markers. The amniocentesis rate was 334 of 1030 (32.4%), and it increased with the number of sonographic markers noted (0 = 29.9%, 1 = 60.2%, 2 = 70.6%, 3 or more = 80%). Genetic ultrasonography is highly effective in identifying chromosomally abnormal fetuses in a community-based practice.

Experimental and theoretical waveforms of Rayleigh waves generated by a thermoelastic laser line source.

An analytical model has been developed for the generation of surface acoustic (Rayleigh) waves in an isotropic solid by a thermoelastic laser line source. For a Gaussian light intensity profile, this model leads to an expression in closed form for the normal surface displacement of the Rayleigh wave either in the near field or in the far field domain. Quantitative agreement has been found for experiments carried out with an interferometric optical probe on a duraluminum plate.