In-plane ESPI with unlimited angle of view for multi-object rotation angle determination.

The in-plane electronic speckle pattern interferometry (ESPI), implemented in a Michelson stellar interferometer-like configuration, offers high sensitivity and dynamic measurement. However, its limited angle of view (AOV) remains a major challenge for the rotation angle determination of multiple objects. In this Letter, we analyze the main factors that influence the AOV of the in-plane ESPI and propose an "image transmitting" approach to enlarge the AOV. With the aid of a folded dual-telescope imaging system, we develop an AOV-unlimited interferometer that can determine multi-object rotation angles in real time. The practicability of the interferometer is demonstrated by the application in real-time measuring of the rotation angles of the disks within a 2D granular system.

Efficient Evaluation of Concrete Fracture Surface Roughness Using Fringe Projection Technology.

The evaluation of concrete surface roughness is crucial in the field of civil engineering. The purpose of this study is to propose a no-contact and efficient method for the measurement of the roughness of concrete fracture surfaces based on fringe-projection technology. A simple phase-correction method using one additional strip image is presented for the phase unwrapping to improve the measurement efficiency and accuracy. The experimental results indicate that the measuring error for plane height is less than 0.1mm, and the relative accuracy for measuring a cylindrical object is about 0.1%, meeting the requirements for concrete fracture-surface measurement. On this basis, three-dimensional reconstructions were carried out on various concrete fracture surfaces to evaluate the roughness. The results reveal that the surface roughness (R) and fractal dimension (D) decrease as the concrete strength increases or the water-to-cement ratio decreases, consistent with previous studies. In addition, compared with the surface roughness, the fractal dimension is more sensitive to the change in concrete surface shape. The proposed method is effective for detecting concrete fracture-surface features.

Biomimetic metal-organic frameworks as protective scaffolds for live-virus encapsulation and vaccine stabilization.

The invaluable health, economic and social impacts of vaccination are hard to exaggerate. The ability to stabilize vaccines is urgently required for their equitable distribution without the dependence on the 'cold-chain' logistics. Herein, for the first time we report biomimetic-mineralization of live-viral vaccines using metal-organic frameworks (MOFs) to enhance their storage stability from days to months. Applying ZIF-8 and aluminium fumarate (Alfum), the Newcastle Disease Virus (NDV) V4 strain and Influenza A WSN strain were encapsulated with remarkable retention of their viral titre. The ZIF-8@NDV, ZIF-8@WSN and Alfum@WSN composites were validated for live-virus recovery using a tissue culture infectious dose (TCID50) assay. With the objective of long-term stabilization, we developed a novel, trehalose (T) and skim milk (SM) stabilized, freeze-dried MOF@Vaccine composite, ZIF-8@NDV+T/SM. The thermal stability of this composite was investigated and compared with the control NDV and non-encapsulated, freeze-dried NDV+T/SM composite at 4 °C, RT, and 37 °C over a period of 12 weeks. We demonstrate the fragility of the control NDV vaccine which lost all viability at RT and 37°C by 12 and 4 weeks, respectively. Comparing the freeze-dried counterparts, the MOF encapsulated ZIF-8@NDV+T/SM demonstrated significant enhancement in stability of the NDV+T/SM composite especially at RT and 37 °C upto 12 weeks. STATEMENT OF SIGNIFICANCE: Vaccination is undoubtedly one of the most effective medical interventions, saving millions of lives each year. However, the requirement of 'cold-chain' logistics is a major impediment to widespread immunization. Live viral vaccines (LVVs) are widely used vaccine types with proven efficacy and low cost. Nonetheless, their complex composition increases their susceptability to thermal stress. Several LVV thermostabilization approaches have been investigated, including their complex engineering and the facile addition of stabilizers. Still, the lack of a universal approach urgently requires finding a stabilization technique especially when additives alone may not be sufficient. Herein, we demonstrate MOF biomimetic-mineralization technology to encapsulate LVVs developing an optimised composite which significantly preserves vaccines without refrigeration for extended periods of time.

Interferon Signaling in Chickens Plays a Crucial Role in Inhibiting Influenza Replication in DF1 Cells.

Influenza A viruses (IAV) pose a constant threat to human and poultry health. Of particular interest are the infections caused by highly pathogenic avian influenza (HPAI) viruses, such as H5N1, which cause significant production issues. In response to influenza infection, cells activate immune mechanisms that lead to increased interferon (IFN) production. To investigate how alterations in the interferon signaling pathway affect the cellular response to infection in the chicken, we used CRISPR/Cas9 to generate a chicken cell line that lacks a functional the type I interferon receptor (IFNAR1). We then assessed viral infections with the WSN strain of influenza. Cells lacking a functional IFNAR1 receptor showed reduced expression of the interferon stimulated genes (ISG) such as Protein Kinase R (PKR) and Myxovirus resistance (Mx) and were more susceptible to viral infection with WSN. We further investigated the role or IFNAR1 on low pathogenicity avian influenza (LPAI) strains (H7N9) and a HPAI strain (H5N1). Intriguingly, Ifnar-/- cells appeared more resistant than WT cells when infected with HPAI virus, potentially indicating a different interaction between H5N1 and the IFN signaling pathway. Our findings support that ChIFNAR1 is a key component of the chicken IFN signaling pathway and these data add contributions to the field of host-avian pathogen interaction and innate immunity in chickens.

Large Deformation and Energy Absorption Behaviour of Perforated Hollow Sphere Structures under Quasi-Static Compression.

Hollow sphere structures with perforations (PHSSs) in three different arrangements (simple cubic (SC), body-centred cubic (BCC), and face-centred cubic (FCC)) were fabricated through three-dimensional (3D) printing, and the mechanical behaviours of these PHSSs under quasi-static compression were investigated experimentally and numerically. The results indicated that under uniaxial compression, the PHSSs mainly undergo three stages, i.e., a linear elastic stage, a large deformation or plateau stage, and a densification stage. During the stage of large deformation, the SC and BCC PHSSs experience a preliminary compaction sub-stage after layer-by-layer buckling, while for the FCC PHSS, layer-by-layer collapse and compaction are the dominant deformation behaviours. A numerical simulation was employed to study the mechanical properties of PHSSs with different geometric parameters under quasi-static compression and to explore the effect of the wall thickness, hole diameter, and sphere arrangement on the first peak stress, plateau stress, and specific energy absorption (SEA) of the PHSSs. The results reveal that the geometric parameters have a significant impact on the large deformation behaviour and energy absorption capacity of PHSSs. The presented PHSS is also proven to be much lighter than traditional metallic hollow sphere structure (MHSS) and has higher specific strength and SEA.

Refractive three-dimensional reconstruction for underwater stereo digital image correlation.

Measuring the three-dimensional (3D) deformation of submerged objects through different media with the stereo digital image correlation (stereo-DIC) method involves the refractive optical imaging problem where the non-linear transmission of light is induced by a change of medium density. The problem invalidates the underlying single viewpoint assumption of the perspective model in regular stereo-DIC, thereby resulting in erroneous measurements of 3D shape and deformation. In this work, we propose a refractive stereo-DIC method that overcomes the problem by considering light refraction in 3D reconstruction. We formulate a full refractive reconstruction geometry description based on Snell's law of flat refraction and the regular triangulation. This allows the true shape to be effectively reconstructed by tracing and establishing the refracted ray-paths based on the regular 3D reconstruction, without reformulating the camera model and image formation. The refractive stereo-DIC is finally established by integrating the refractive 3D reconstruction into the regular DIC framework for measuring accurate 3D shape and deformation of submerged objects. We experiment the proposed approach with underwater 3D shape and deformation measurements. Both results prove its feasibility and correctness, further heralding our approach as a flexible solution that could readily extend the stereo-DIC to fluid-immersed 3D deformation characterization.

Analysis of the Evolution of Pandemic Influenza A(H1N1) Virus Neuraminidase Reveals Entanglement of Different Phenotypic Characteristics.

The influenza A virus (IAV) neuraminidase (NA) is essential for virion release from cells and decoy receptors and an important target of antiviral drugs and antibodies. Adaptation to a new host sialome and escape from the host immune system are forces driving the selection of mutations in the NA gene. Phylogenetic analysis shows that until 2015, 16 amino acid substitutions in NA became fixed in the virus population after introduction in the human population of the pandemic IAV H1N1 (H1N1pdm09) in 2009. The accumulative effect of these substitutions, in the order in which they appeared, was analyzed using recombinant proteins and viruses in combination with different functional assays. The results indicate that NA activity did not evolve to a single optimum but rather fluctuated within a certain bandwidth. Furthermore, antigenic and enzymatic properties of NA were intertwined, with several residues affecting multiple properties. For example, the substitution K432E in the second sialic acid binding site, next to the catalytic site, was shown to affect catalytic activity, substrate specificity, and the pH optimum for maximum activity. This substitution also altered antigenicity of NA, which may explain its selection. We propose that the entanglement of NA phenotypes may be an important determining factor in the evolution of NA.IMPORTANCE Since its emergence in 2009, the pandemic H1N1 influenza A virus (IAV) has caused significant disease and mortality in humans. IAVs contain two envelope glycoproteins, the receptor-binding hemagglutinin (HA) and the receptor-destroying neuraminidase (NA). NA is essential for virion release from cells and decoy receptors, is an important target of antiviral drugs, and is increasingly being recognized as an important vaccine antigen. Not much is known, however, about the evolution of this protein upon the emergence of the novel pandemic H1N1 virus, with respect to its enzymatic activity and antigenicity. By reconstructing the evolutionary path of NA, we show that antigenic and enzymatic properties of NA are intertwined, with several residues affecting multiple properties. Understanding the entanglement of NA phenotypes will lead to better comprehension of IAV evolution and may help the development of NA-based vaccines.

Investigating the Interaction between Negative Strand RNA Viruses and Their Hosts for Enhanced Vaccine Development and Production.

The current pandemic has highlighted the ever-increasing risk of human to human spread of zoonotic pathogens. A number of medically-relevant zoonotic pathogens are negative-strand RNA viruses (NSVs). NSVs are derived from different virus families. Examples like Ebola are known for causing severe symptoms and high mortality rates. Some, like influenza, are known for their ease of person-to-person transmission and lack of pre-existing immunity, enabling rapid spread across many countries around the globe. Containment of outbreaks of NSVs can be difficult owing to their unpredictability and the absence of effective control measures, such as vaccines and antiviral therapeutics. In addition, there remains a lack of essential knowledge of the host-pathogen response that are induced by NSVs, particularly of the immune responses that provide protection. Vaccines are the most effective method for preventing infectious diseases. In fact, in the event of a pandemic, appropriate vaccine design and speed of vaccine supply is the most critical factor in protecting the population, as vaccination is the only sustainable defense. Vaccines need to be safe, efficient, and cost-effective, which is influenced by our understanding of the host-pathogen interface. Additionally, some of the major challenges of vaccines are the establishment of a long-lasting immunity offering cross protection to emerging strains. Although many NSVs are controlled through immunisations, for some, vaccine design has failed or efficacy has proven unreliable. The key behind designing a successful vaccine is understanding the host-pathogen interaction and the host immune response towards NSVs. In this paper, we review the recent research in vaccine design against NSVs and explore the immune responses induced by these viruses. The generation of a robust and integrated approach to development capability and vaccine manufacture can collaboratively support the management of outbreaking NSV disease health risks.

Panoramic dual-directional shearography assisted by a bi-mirror.

A panoramic dual-directional shearography system is proposed to simultaneously determine out-of-plane deformation derivatives in two directions and globally inspect the object to be tested. A dichroic filter (DF), a 3CCD camera, and dual-wavelength light are used in the proposed shearography configuration. The dual-wavelength light coupled with the corresponding imaging sensors of the 3CCD camera provides independent color signals and shearograms. Through adjustment of the tilted stereo-angle of the DF, which offers a second wavelength-dependent measurement, an additional independent image-shearing can be introduced into the setup. The auxiliary bi-mirror surrounding the object helps to fully illuminate the object surface and capture it in a single shot. Theoretical analysis and experimental results demonstrated the utility of the system.

Substrate Binding by the Second Sialic Acid-Binding Site of Influenza A Virus N1 Neuraminidase Contributes to Enzymatic Activity.

The influenza A virus (IAV) neuraminidase (NA) protein plays an essential role in the release of virus particles from cells and decoy receptors. The NA enzymatic activity presumably needs to match the activity of the IAV hemagglutinin (HA) attachment protein and the host sialic acid (SIA) receptor repertoire. We analyzed the enzymatic activities of N1 NA proteins derived from avian (H5N1) and human (H1N1) IAVs and analyzed the role of the second SIA-binding site, located adjacent to the conserved catalytic site, therein. SIA contact residues in the second SIA-binding site of NA are highly conserved in avian, but not human, IAVs. All N1 proteins preferred cleaving α2,3- over α2,6-linked SIAs even when their corresponding HA proteins displayed a strict preference for α2,6-linked SIAs, indicating that the specificity of the NA protein does not need to fully match that of the corresponding HA protein. NA activity was affected by substitutions in the second SIA-binding site that are observed in avian and human IAVs, at least when multivalent rather than monovalent substrates were used. These mutations included both SIA contact residues and residues that do not directly interact with SIA in all three loops of the second SIA-binding site. Substrate binding via the second SIA-binding site enhanced the catalytic activity of N1. Mutation of the second SIA-binding site was also shown to affect virus replication in vitro Our results indicate an important role for the N1 second SIA-binding site in binding to and cleavage of multivalent substrates.IMPORTANCE Avian and human influenza A viruses (IAVs) preferentially bind α2,3- and α2,6-linked sialic acids (SIAs), respectively. A functional balance between the hemagglutinin (HA) attachment and neuraminidase (NA) proteins is thought to be important for host tropism. What this balance entails at the molecular level is, however, not well understood. We now show that N1 proteins of both avian and human viruses prefer cleaving avian- over human-type receptors although human viruses were relatively better in cleavage of the human-type receptors. In addition, we show that substitutions at different positions in the second SIA-binding site found in NA proteins of human IAVs have a profound effect on binding and cleavage of multivalent, but not monovalent, receptors and affect virus replication. Our results indicate that the HA-NA balance can be tuned via modification of substrate binding via this site and suggest an important role of the second SIA-binding site in host tropism.

Kinetic analysis of the influenza A virus HA/NA balance reveals contribution of NA to virus-receptor binding and NA-dependent rolling on receptor-containing surfaces.

Interactions of influenza A virus (IAV) with sialic acid (SIA) receptors determine viral fitness and host tropism. Binding to mucus decoy receptors and receptors on epithelial host cells is determined by a receptor-binding hemagglutinin (HA), a receptor-destroying neuraminidase (NA) and a complex in vivo receptor-repertoire. The crucial but poorly understood dynamics of these multivalent virus-receptor interactions cannot be properly analyzed using equilibrium binding models and endpoint binding assays. In this study, the use of biolayer interferometric analysis revealed the virtually irreversible nature of IAV binding to surfaces coated with synthetic sialosides or engineered sialoglycoproteins in the absence of NA activity. In addition to HA, NA was shown to be able to contribute to the initial binding rate while catalytically active. Virus-receptor binding in turn contributed to receptor cleavage by NA. Multiple low-affinity HA-SIA interactions resulted in overall extremely high avidity but also permitted a dynamic binding mode, in which NA activity was driving rolling of virus particles over the receptor-surface. Virus dissociation only took place after receptor density of the complete receptor-surface was sufficiently decreased due to NA activity of rolling IAV particles. The results indicate that in vivo IAV particles, after landing on the mucus layer, reside continuously in a receptor-bound state while rolling through the mucus layer and over epithelial cell surfaces driven by the HA-NA-receptor balance. Quantitative BLI analysis enabled functional examination of this balance which governs this dynamic and motile interaction that is expected to be crucial for penetration of the mucus layer and subsequent infection of cells by IAV but likely also by other enveloped viruses carrying a receptor-destroying enzyme in addition to a receptor-binding protein.

Shape measurement with modified phase-shift lateral shearing interferometry illumination and radial basis function.

Three-dimensional shapes of objects were evaluated with modified phase-shift lateral shearing interferometry illumination and radial basis function. A simple optical system was developed to create the fringe pattern based on the Murty interferometer. The phase shift was generated only by moving a plane-parallel plate along an in-plane parallel direction. A novel moving radial basis function method was presented to improve the quality of fringe patterns. And the proper calculation window size was given based on numerical simulation. Three-dimensional shapes of two kinds of objects were determined to verify the feasibility and effectiveness of the proposed method, and the reconstructed height distributions were in good accordance with the referenced data.

Mutation of the Second Sialic Acid-Binding Site, Resulting in Reduced Neuraminidase Activity, Preceded the Emergence of H7N9 Influenza A Virus.

The emergence of the novel influenza A virus (IAV) H7N9 since 2013 has caused concerns about the ability of the virus to spread between humans. Analysis of the receptor-binding properties of the H7 protein of a human isolate revealed modestly increased binding to α2,6 sialosides and reduced, but still dominant, binding to α2,3-linked sialic acids (SIAs) compared to a closely related avian H7N9 virus from 2008. Here, we show that the corresponding N9 neuraminidases (NAs) display equal enzymatic activities on a soluble monovalent substrate and similar substrate specificities on a glycan array. In contrast, solid-phase activity and binding assays demonstrated reduced specific activity and decreased binding of the novel N9 protein. Mutational analysis showed that these differences resulted from substitution T401A in the 2nd SIA-binding site, indicating that substrate binding via this site enhances NA catalytic activity. Substitution T401A in the novel N9 protein appears to functionally mimic the substitutions that are found in the 2nd SIA-binding site of NA proteins of avian-derived IAVs that became human pandemic viruses. Our phylogenetic analyses show that substitution T401A occurred prior to substitutions in hemagglutinin (HA), causing the altered receptor-binding properties mentioned above. Hence, in contrast to the widespread assumption that such changes in NA are obtained only after acquisition of functional changes in HA, our data indicate that mutations in the 2nd SIA-binding site may have enabled and even driven the acquisition of altered HA receptor-binding properties and may have contributed to the spread of the novel H7N9 viruses.IMPORTANCE Novel H7N9 IAVs continue to cause human infections and pose an ongoing public health threat. Here, we show that their N9 proteins display reduced binding to and lower enzymatic activity against multivalent substrates, resulting from mutation of the 2nd sialic acid-binding site. This mutation preceded and may have driven the selection of substitutions in H7 that modify H7 receptor-binding properties. Of note, all animal IAVs that managed to cross the host species barrier and became human viruses carry mutated 2nd sialic acid-binding sites. Screening of animal IAVs to monitor their potential to cross the host species barrier should therefore focus not only on the HA protein, but also on the functional properties of NA.

Identification of Residues That Affect Oligomerization and/or Enzymatic Activity of Influenza Virus H5N1 Neuraminidase Proteins.

UNLABELLED: Influenza A virus (IAV) attachment to and release from sialoside receptors is determined by the balance between hemagglutinin (HA) and neuraminidase (NA). The molecular determinants that mediate the specificity and activity of NA are still poorly understood. In this study, we aimed to design the optimal recombinant soluble NA protein to identify residues that affect NA enzymatic activity. To this end, recombinant soluble versions of four different NA proteins from H5N1 viruses were compared with their full-length counterparts. The soluble NA ectodomains were fused to three commonly used tetramerization domains. Our results indicate that the particular oligomerization domain used does not affect the Km value but may affect the specific enzymatic activity. This particularly holds true when the stalk domain is included and for NA ectodomains that display a low intrinsic ability to oligomerize. NA ectodomains extended with a Tetrabrachion domain, which forms a nearly parallel four-helix bundle, better mimicked the enzymatic properties of full-length proteins than when other coiled-coil tetramerization domains were used, which probably distort the stalk domain. Comparison of different NA proteins and mutagenic analysis of recombinant soluble versions thereof resulted in the identification of several residues that affected oligomerization of the NA head domain (position 95) and therefore the specific activity or sialic acid binding affinity (Km value; positions 252 and 347). This study demonstrates the potential of using recombinant soluble NA proteins to reveal determinants of NA assembly and enzymatic activity. IMPORTANCE: The IAV HA and NA glycoproteins are important determinants of host tropism and pathogenicity. However, NA is relatively understudied compared to HA. Analysis of soluble versions of these glycoproteins is an attractive way to study their activities, as they are easily purified from cell culture media and applied in downstream assays. In the present study, we analyzed the enzymatic activity of different NA ectodomains with three commonly used tetramerization domains and compared them with full-length NA proteins. By performing a mutagenic analysis, we identified several residues that affected NA assembly, activity, and/or substrate binding. In addition, our results indicate that the design of the recombinant soluble NA protein, including the particular tetramerization domain, is an important determinant for maintaining the enzymatic properties within the head domain. NA ectodomains extended with a Tetrabrachion domain better mimicked the full-length proteins than when the other tetramerization domains were used.

Effects of zearalenone-diet on expression of ghrelin and PCNA genes in ovaries of post-weaning piglets.

Numerous reports have provided evidence that zearalenone (ZEN) can increase the weight of genital organs. These findings have been confirmed by many studies in which the ghrelin gene was expressed in the ovary and was implicated in the control of cells in reproductive tissues. The proliferating cell nuclear antigen (PCNA) is an important marker of cell proliferation. The present study investigates the effects of a ZEN-treated diet on the development of ovaries in post-weaning piglets by the detection of ghrelin and PCNA protein and relative abundance of mRNA using optical microscopy, immunohistochemistry and quantitative real-time (qRT-PCR). A total of 20 piglets (Duroc × Landrace × Yorkshire) weaned at 28 d, with an average body weight of 8.74 ± 0.26 kg (P=0.919) were used in this study. Piglets in the control group (n=10) were fed a normal basal diet, and those in the treatment group (n=10) were fed a diet containing ZEN (1.04 mg/kg), for 35 d. The proportion of growing follicles and diameter of the largest growing follicle in ovaries were greater in piglets fed the diet with ZEN. The total integrated optical densities of protein and mRNA of ghrelin and PCNA were greater with the feeding of the ZEN-treatment diet. The results suggested that 1.04 mg/kg ZEN could promote the autocrine action or expression of the ghrelin gene in piglet ovary, and further accelerate the development of ovaries (follicles).

Rapid Emergence of Highly Pathogenic Avian Influenza Subtypes from a Subtype H5N1 Hemagglutinin Variant.

In 2014, novel highly pathogenic avian influenza A H5N2, H5N5, H5N6, and H5N8 viruses caused outbreaks in Asia, Europe, and North America. The H5 genes of these viruses form a monophyletic group that evolved from a clade 2.3.4 H5N1 variant. This rapid emergence of new H5Nx combinations is unprecedented in the H5N1 evolutionary history.

Transmissible gastroenteritis virus infection induces cell cycle arrest at S and G2/M phases via p53-dependent pathway.

p53 signaling pathway plays an important role in the regulation of cell cycle. Our previous studies have demonstrated that TGEV infection induces the activation of p53 signaling pathway. In this study we investigated the effects of TGEV infection on the cell cycle of host cells and the roles of p53 activation in this process. The results showed that TGEV infection induced cell cycle arrest at S and G2/M phases in both asynchronous and synchronized PK-15 and ST cells, while UV-inactivated TGEV lost the ability of induction of cell cycle arrest. TGEV infection promoted p21 accumulation, down-regulated cell cycle-regulatory proteins cyclins B1, cdc2, cdk2 and PCNA. Further studies showed that inhibition of p53 signaling could attenuate the TGEV-induced S- and G2/M-phase arrest by reversing the expression of p21 and corresponding cyclin/cdk. In addition, TGEV infection of the cells synchronized in various stages of cell cycle showed that viral genomic RNA and subgenomic RNA, and virus titer were higher in the cells released from S-phase- or G2/M phase-synchronized cells than that in the cells released from the G0/G1 phase-synchronized or asynchronous cells after 18h p.i. Taken together, our data suggested that TGEV infection induced S and G2/M phase arrest in host cells, which might provide a favorable condition for viral replication.

Calibration of revolution axis for 360 deg surface measurement.

Two methods are proposed to calibrate the revolution axis of a 360 deg, multiview fringe projection system for surface measurement. The first method is based on minimizing the distance between calculated and measured points; the second method is based on minimizing the difference between thus obtained vectors. Both are able to retrieve the revolution axis of a turntable, which is then used to transform surface patches measured at different viewing angles to a common coordinate. In the point-based method, a nonlinear minimization problem has to be solved by the Levenberg-Marquardt algorithm; in the vector-based method, the minimization problem is resolved into several linear equations, and an analytic solution is obtained efficiently. Results of simulation and experiments show that the error of calibration can be less than 0.05 deg for the axis's orientation and 0.3 mm for the axis's position (a point on the axis), which is about 0.1% of the measured volume.

Transmissible gastroenteritis virus infection induces cell apoptosis via activation of p53 signalling.

Transmissible gastroenteritis virus (TGEV) infection induced apoptosis in several cell lines in vitro. Our previous studies demonstrated that TGEV could activate FasL- and mitochondria-mediated pathways to induce apoptosis in PK-15 cells. In this study, we investigated the regulation of p53 and p38 mitogen-activated protein kinases (MAPK) signalling pathways in the interaction of TGEV with host cells. We observed that TGEV infection decreased p300/CBP, downregulated MDM2 and promoted p53 phosphorylation at serines 15, 20 and 46, resulting in accumulation and activation of p53 in PK-15 cells. TGEV infection induced the transient activation of p38 MAPK in the early phase of inoculation and constant activation in the later phase of infection. However, UV-irradiated TGEV did not promote the activation of p53 and p38 MAPK in the later phase, whereas it only triggered the transient activation of p38 MAPK in the early phase. Blocking of p53 activation significantly inhibited the occurrence of apoptosis through suppressing the TGEV-induced FasL expression, Bcl-2 reduction, Bax and cytochrome c redistribution, while inhibition of p38 activity moderately blocked apoptosis induction and partly attenuated the accumulation and activation of p53. However, inhibition of p38 and p53 activity had no significant effects on viral gene transcription at 12 and 24 h post-infection. Taken together, these results demonstrated that TGEV infection promoted the activation of p38 MAPK and p53 signalling, and p53 signalling might play a dominant role in the regulation of cell apoptosis. These findings provide new insights into the function of p53 and p38 MAPK in the interaction of TGEV with host cells.

Single-shot color fringe projection for three-dimensional shape measurement of objects with discontinuities.

A simple but effective fringe projection profilometry is proposed to measure 3D shape by using one snapshot color sinusoidal fringe pattern. One color fringe pattern encoded with a sinusoidal fringe (as red component) and one uniform intensity pattern (as blue component) is projected by a digital video projector, and the deformed fringe pattern is recorded by a color CCD camera. The captured color fringe pattern is separated into its RGB components and division operation is applied to red and blue channels to reduce the variable reflection intensity. Shape information of the tested object is decoded by applying an arcsine algorithm on the normalized fringe pattern with subpixel resolution. In the case of fringe discontinuities caused by height steps, or spatially isolated surfaces, the separated blue component is binarized and used for correcting the phase demodulation. A simple and robust method is also introduced to compensate for nonlinear intensity response of the digital video projector. The experimental results demonstrate the validity of the proposed method.