Diffusion-Weighted MRI of the Liver in Patients With Chronic Liver Disease: A Comparative Study Between Different Fitting Approaches and Diffusion Models.

BACKGROUND: Diffusion-weighted imaging (DWI) has been considered for chronic liver disease (CLD) characterization. Grading of liver fibrosis is important for disease management. PURPOSE: To investigate the relationship between DWI's parameters and CLD-related features (particularly regarding fibrosis assessment). STUDY TYPE: Retrospective. SUBJECTS: Eighty-five patients with CLD (age: 47.9 ± 15.5, 42.4% females). FIELD STRENGTH/SEQUENCE: 3-T, spin echo-echo planar imaging (SE-EPI) with 12 b-values (0-800 s/mm2 ). ASSESSMENT: Several models statistical models, stretched exponential model, and intravoxel incoherent motion were simulated. The corresponding parameters (Ds , σ, DDC, α, f, D, D*) were estimated on simulation and in vivo data using the nonlinear least squares (NLS), segmented NLS, and Bayesian methods. The fitting accuracy was analyzed on simulated Rician noised DWI. In vivo, the parameters were averaged from five central slices entire liver to compare correlations with histological features (inflammation, fibrosis, and steatosis). Then, the differences between mild (F0-F2) or severe (F3-F6) groups were compared respecting to statistics and classification. A total of 75.3% of patients used to build various classifiers (stratified split strategy and 10-folders cross-validation) and the remaining for testing. STATISTICAL TESTS: Mean squared error, mean average percentage error, spearman correlation, Mann-Whitney U-test, receiver operating characteristic (ROC) curve, area under ROC curve (AUC), sensitivity, specificity, accuracy, precision. A P-value <0.05 was considered statistically significant. RESULTS: In simulation, the Bayesian method provided the most accurate parameters. In vivo, the highest negative significant correlation (Ds , steatosis: r = -0.46, D*, fibrosis: r = -0.24) and significant differences (Ds , σ, D*, f) were observed for Bayesian fitted parameters. Fibrosis classification was performed with an AUC of 0.92 (0.91 sensitivity and 0.70 specificity) with the aforementioned diffusion parameters based on the decision tree method. DATA CONCLUSION: These results indicate that Bayesian fitted parameters may provide a noninvasive evaluation of fibrosis with decision tree. EVIDENCE LEVEL: 1 TECHNICAL EFFICACY: Stage 1.

Computed torque control and force analysis for mechanical leg with variable rotation axis powered by servo pneumatic muscle.

It is difficult for a humanoid leg driven by two groups of antagonistic pneumatic muscles (PMs) to achieve a flexible humanoid gait, and its inherent strong coupling nonlinear characteristics make it hard to achieve good tracking performance in a large range of motion. Therefore, a four-bar linkage bionic knee joint structure with a variable axis and a double closed-loop servo position control strategy based on computed torque control are designed to improve anthropomorphic characteristics and the dynamic performance of the bionic mechanical leg powered by servo pneumatic muscle (SPM). Firstly, the relationship between the joint torque, the initial jump angle and the bounce height of the mechanical leg is established, and then we design a double-joint PM bionic mechanical leg containing a four-bar linkage mechanism of the knee joint. Secondly, a cascade position control strategy is developed, which consists of the outer position loop and the inner contraction force loop, and the mapping relationship is designed between joint torque and antagonistic PM contraction force. Finally, we further project bounce action timing of mechanical leg to realize the periodic jumping movement of the mechanical leg, and simulation and physical experiments of the real-style machine platform have been provided to demonstrate the effectiveness of the designed SPM controller.

In-vivo programmable acoustic manipulation of genetically engineered bacteria.

Acoustic tweezers can control target movement through the momentum interaction between an acoustic wave and an object. This technology has advantages over optical tweezers for in-vivo cell manipulation due to its high tissue penetrability and strong acoustic radiation force. However, normal cells are difficult to acoustically manipulate because of their small size and the similarity between their acoustic impedance and that of the medium. In this study, we use the heterologous expression of gene clusters to generate genetically engineered bacteria that can produce numerous sub-micron gas vesicles in the bacterial cytoplasm. We show that the presence of the gas vesicles significantly enhances the acoustic sensitivity of the engineering bacteria, which can be manipulated by ultrasound. We find that by employing phased-array-based acoustic tweezers, the engineering bacteria can be trapped into clusters and manipulated in vitro and in vivo via electronically steered acoustic beams, enabling the counter flow or on-demand flow of these bacteria in the vasculature of live mice. Furthermore, we demonstrate that the aggregation efficiency of engineering bacteria in a tumour is improved by utilizing this technology. This study provides a platform for the in-vivo manipulation of live cells, which will promote the progress of cell-based biomedical applications.

Flexible Ultrasound Transducer With Embedded Optical Shape Sensing Fiber for Biomedical Imaging Applications.

Flexible ultrasound transducers (FUTs), capable of conforming to irregular surfaces, have become a research hotspot in the field of medical imaging. With these transducers, high-quality ultrasound images can be obtained only if strict design criteria are fulfilled. Moreover, the relative positions of array elements must be determined, which are important for ultrasound beamforming and image reconstruction. These two major characteristics present great challenges to the design and fabrication of FUTs compared to that for traditional rigid probes. In this study, an optical shape-sensing fiber was embedded into a 128-element flexible linear array transducer to acquire the real-time relative positions of array elements to produce high-quality ultrasound images. Minimum concave and convex bend diameters of approximately 20 and 25 mm, respectively, were achieved. The transducer was flexed 2000 times, and yet no obvious damage was observed. Stable electrical and acoustic responses confirmed its mechanical integrity. The developed FUT exhibited an average center frequency of 6.35 MHz, and average -6-dB bandwidth of 69.2%. The array profile and element positions measured by the optic shape-sensing system were instantly transferred to the imaging system. Phantom experiments for both spatial resolution and contrast-to-noise ratio proved that FUTs can maintain satisfactory imaging capability despite bending to sophisticated geometries. Finally, color Doppler images and Doppler spectra of the peripheral arteries of healthy volunteers were obtained in real time.

Circular array transducer based-photoacoustic/ultrasonic endoscopic imaging with tunable ring-beam excitation.

Photoacoustic/ultrasound endoscopic imaging is regarded as an effective method to achieve accurate detection of intestinal disease by offering both the functional and structural information, simultaneously. Compared to the conventional endoscopy with single transducer and laser spot for signal detection and optical excitation, photoacoustic/ultrasound endoscopic probe using circular array transducer and ring-shaped laser beam avoids the instability brought by the mechanical scanning point-to-point, offering the dual-modality imaging with high accuracy and efficiency. Meanwhile, considering the complex morphological environments of intestinal tracts in clinics, developing the probe having sufficient wide imaging distance range is especially important. In this work, we develop a compact circular photoacoustic/ultrasonic endoscopic probe, using the group of fiber, lens and home-made axicon, to generate relatively concentrated ring-shaped laser beam for 360° excitation with high efficiency. Furthermore, the laser ring size can be tuned conveniently by changing the fiber-lens distance to ensure the potential applicability of the probe in various and complex morphological environments of intestines. Phantom experimental results demonstrate imaging distance range wide enough to cover from 12 mm to 30 mm. In addition, the accessibility of the photoacoustic signals of molecular probes in ex vivo experiments at the tissue depth of 7 mm using excitation energy of 5 mJ has also been demonstrated, showing a high optical excitation efficiency of the probe.

Sonodynamic Therapy With Concentric Ultrasound Imaging Array for Precision Theranostics for Atherosclerotic Plaque.

Atherosclerotic cardiovascular disease is a major cause of human disability and mortality. Our previous study demonstrated the safety and efficacy of sonodynamic therapy (SDT) on atherosclerotic plaques. However, traditional single-element therapeutic transducer has single acoustic field, and positioning therapeutic and imaging transducers in the same position is difficult during ultrasound imaging-guided SDT. Continuously changing the position of transducers to intervene lesions in different positions is required, increasing the difficulty of treatment. Thus, an SDT device with precise theranostics is required. Therefore, we designed and fabricated a "concentric ultrasound transducer for theranostics" (CUST-T), comprising a central 8-MHz linear array transducer for ultrasound imaging, and a peripheral 1-MHz hollow two-dimensional (2-D) planar array transducer for generating phased-array focused ultrasound (PAFUS). The CUST-T exhibited high imaging resolution at a distance of up to 20 mm from the transducer and could generate a personalized complex PAFUS acoustic field to match various lesions. In vitro biomedical results showed that PAFUS-SDT induced RAW264.7-derived foam cell apoptosis leading to a targeting field apoptotic rate 4.36-6.24 times that of the nontargeting field and the significant apoptotic region was consistent with the PAFUS acoustic field. In vivo, PAFUS-SDT guided by ultrasound imaging significantly increased the lumen area ( ) and collagen level ( ), whereas the wall thickness ( ) and lipid content ( ) of rabbit femoral artery were reduced. In conclusion, CUST-T provided image guidance sufficient for accurate SDT for atherosclerotic plaques in peripheral arteries and could be applied in clinical practice.

Endoscopic Ultrasound Localization Microscopy for the Evaluation of the Microvasculature of Gastrointestinal Tract Tumors in Rabbits.

OBJECTIVE: The morphological and hemodynamic characterization of the microvascular network around the gastrointestinal (GI) tract can be of significant clinical value for the early diagnosis and treatment of GI tract cancer. Ultrasound localization microscopy (ULM) imaging has been demonstrated to be capable of resolving the microvascular network. However, the endoscopic application of ULM imaging techniques is still unknown. In this study, an endoscopic ultrasound localization microscopy (e-ULM) imaging technique was developed to evaluate the changes of microvasculature during GI tract tumor growth. METHODS: A customized circular array transducer (center frequency: 6.8 MHz) and the coherent diverging wave compounding method were used to generate B-mode images. Spatiotemporal singular value decomposition processing was used to eliminate the background signals before signal localizations. The centroids of spatially isolated signals were localized and summed to generate the final super-resolution image. RESULTS: The final microvasculature map of a rabbit GI tract tumor reveals that e-ULM can be used to surpass the diffraction limit in traditional endoscopic ultrasound (EUS) imaging. Furthermore, it is observed that data from different stages of tumor growth exhibit significant differences in microvascular pattern and density. CONCLUSION: Our study demonstrated the implementation and application of an in vivo e-ULM imaging technique for the evaluation of the microvasculature of GI tumors. SIGNIFICANCE: The efficient e-ULM imaging technique shows potential for use in the detection of GI tract tumor microcirculation changes and subsequent diagnosis of GI tract cancer.

3D Acoustic Manipulation of Living Cells and Organisms Based On 2D Array.

Flexible manipulation techniques for living cells and organisms are extremely useful tools for fundamental biomedical and life science research. Acoustic tweezers, which permit non-contact, label-free manipulation, are particularly suited to micromanipulation tasks as they provide a large acoustic radiation force and can be applied in various media. Here, we describe the design and fabrication of a 3 MHz, 64-element (8 × 8), 2D planar ultrasound array that realizes the multidimensional translation, rotation, orientation, and levitation of living cells and organisms. The focusing vortex and twin fields are generated using the holographic acoustic elements framework method. We demonstrate that the eggs and larvae of brine shrimp can be translated along a preset trajectory by controlling the central position of the vortex. By multiplexing counterclockwise vortices, clockwise vortices, and twin trap fields in a time sequence, the rotation direction of the shrimp eggs can be switched in real time, while non-spherical larvae can be reoriented. Moreover, the reflection of the acoustic beam can lift eggs and larvae from the bottom of the culture dish and further manipulate them in the vertical and horizontal directions. Additionally, we present quantitative analyses of the shrimp-egg rotation frequency with respect to the focal depths, topological charges of the vortex, and excitation voltages. These results indicate that acoustic tweezers based on 2D matrix arrays can realize complex and selective manipulation of living cells and organisms, thereby demonstrating their value for advancing research in the fields of cell assembly, tissue engineering, and micro-robot driving.

Design and Fabrication of a High-Frequency Microconvex Array Transducer for Small Animals Imaging.

High-frequency convex array transducer, featuring both high spatial resolution and wide field of view, has been successfully developed for ophthalmic imaging. To further expand its application range to small animals' imaging, this work develops a high-frequency microconvex array transducer possessing smaller aperture size and wider scanning angle. This transducer featured 128 array elements arranged in a curvilinear 2-2 piezoelectric composite configuration, yielding a maximum view angle of 97.8°. The array was composed of two front matching layers, a nonconductive backing layer, and a customized flexible circuit that electrically connected array elements to coaxial cables. The center frequency and the -6-dB fractional bandwidth were about 18.14 MHz and 69.15%, respectively. The image of a tungsten wire phantom resulted in approximately 62.9- [Formula: see text] axial resolution and 121.3- [Formula: see text] lateral resolution. The image of the whole kidney of a rat as well as its internal arteries was acquired in vivo, demonstrating the imaging capability of the proposed high-frequency microconvex array transducers for small animals' imaging applications.

1.5-Dimensional Circular Array Transducer for In Vivo Endoscopic Ultrasonography.

OBJECTIVE: Traditional endoscopic ultrasonography (EUS), which uses one-dimensional (1-D) curvilinear or radial/circular transducers, cannot achieve dynamic elevational focusing, and the slice thickness is not sufficient. The purpose of this study was to design and fabricate a 1.5-dimensional (1.5-D) circular array transducer to achieve dynamic elevational focusing in EUS in vivo. METHODS: An 84 × 5 element 1.5-D circular array transducer was successfully developed and characterized in this study. It was fabricated with PZT-5H 1-3 composite that attained a high-electromechanical coupling factor and low-acoustic impedance. The acoustic field distribution was measured with different transmission modes to validate the 1.5-D elevational beam focusing capability. The imaging performance of the 84 × 5 element 1.5-D circular array transducer was evaluated by two wire phantoms, an agar-based cyst phantom, an ex vivo swine pancreas, and an in vivo rhesus macaque rectum based on multifocal ray-line imaging method with five-row elevational beam steering. RESULTS: It was demonstrated that the transducer exhibited a central frequency of 6.47 MHz with an average bandwidth of 50%, a two-way insertion loss of 23 dB, and crosstalk of <-26 dB around the center frequency. CONCLUSION: Dynamic elevational focusing and the enhancement of the slice thickness in EUS were obtained with a 1.5-D circular array transducer. SIGNIFICANCE: This study promotes the development of multirow and two-dimensional array EUS probes for a more precise clinical diagnosis and treatment.

Self-Navigated 3D Acoustic Tweezers in Complex Media Based on Time Reversal.

Acoustic tweezers have great application prospects because they allow noncontact and noninvasive manipulation of microparticles in a wide range of media. However, the nontransparency and heterogeneity of media in practical applications complicate particle trapping and manipulation. In this study, we designed a 1.04 MHz 256-element 2D matrix array for 3D acoustic tweezers to guide and monitor the entire process using real-time 3D ultrasonic images, thereby enabling acoustic manipulation in nontransparent media. Furthermore, we successfully performed dynamic 3D manipulations on multiple microparticles using multifoci and vortex traps. We achieved 3D particle manipulation in heterogeneous media (through resin baffle and ex vivo macaque and human skulls) by introducing a method based on the time reversal principle to correct the phase and amplitude distortions of the acoustic waves. Our results suggest cutting-edge applications of acoustic tweezers such as acoustical drug delivery, controlled micromachine transfer, and precise treatment.

Fiber-Based Clock Synchronization Method for Medical Ultrasound System.

Brain ultrasound has attracted great attention recently due to its noninvasive treatment function for brain diseases. However, ultrasound is still difficult to pass through an intact skull. Phase correction is recognized as an effective method for skull compensation. Half-wavelength pitch transducer is important for the phase correction and, hence, thousands of elements array is required to cover large area human tissue. The clock synchronization between elements is crucial for the phase correction; however, the traditional clock scheme which is designed for 128- or 256-element system is not suitable for thousands of elements. In addition, the clock scheme needs to be magnetic resonance imaging (MRI) compatible since MRI-guided intervention is becoming a routine operation for the brain ultrasound. This study is the first to propose an optical fiber-based clock synchronization method for MRI-guided ultrasound array system. The optical fiber not only distributes the clock but also sets up a link to transmit the data for ultrasound beamformer. The link is full-duplex so both the clock and the data can be transmitted and received simultaneously. The precision of clock synchronization is less than 557 ps when using 50 MHz clock, and the period jitter of the clock is less than 10 ps (rms). Multiple 128- or 256-channel ultrasonic systems can be synchronized, and the error between the channels can be less than 10 ns when using 1-MHz ultrasound transducer. The system can work in an MRI scanning room and communicate with a console via only one fiber. In vivo primate animal study has been achieved, and it has been proven that the proposed clock scheme is suitable for MRI-guided large-scale ultrasound array system.

Transcranial Low-Intensity Pulsed Ultrasound Stimulation Induces Neuronal Autophagy.

Autophagy, or cellular self-digestion, is an essential process for eliminating abnormal protein in mammalian cells. Accumulating evidence indicates that increased neuronal autophagy has a protective effect on neurodegenerative disorders. It has been reported that low-intensity pulsed ultrasound (LIPUS) can noninvasively modulate neural activity in the brain. Yet, the effect of LIPUS on neuronal autophagy is still unclear. The objective of this study was to examine whether LIPUS stimulation could induce neuronal autophagy. Primary neurons were treated by LIPUS with a frequency of 0.68 MHz, a pulse repetition frequency (PRF) of 500 Hz, a spatial peak temporal-average intensities ( [Formula: see text]) of 70 and 165 mW/cm2. Then, the immunofluorescent analysis of LC3B was carried out for evaluating neuronal autophagy. Furthermore, 0.5-MHz LIPUS was noninvasively delivered to the cortex and hippocampus of adult mice ( n = 16 ) with PRF of 500 Hz and [Formula: see text] of 235 mW/cm2. The LC3BII/LC3BI ratio and p62 (autophagic markers) were measured by western blot analysis. In the in vitro study, the expression of LC3B in primary neurons was statistically improved after LIPUS stimulation was implemented for 4 h ( ). With the increase in the irradiation duration or acoustic intensity of LIPUS stimulation, the expression of LC3B in primary neurons was increased. Furthermore, transcranial LIPUS stimulation increased the LC3BII/LC3BI ratio ( ) and decreased the expression of p62 ( ) in the cortex and hippocampus. We concluded that LIPUS provides a safe and capable tool for activating neuronal autophagy in vitro and in vivo.

A PMN-PT Composite-Based Circular Array for Endoscopic Ultrasonic Imaging.

Endoscopic ultrasound (EUS), an interventional imaging technology, utilizes a circular array to delineate the cross-sectional morphology of internal organs through the gastrointestinal (GI) track. However, the performance of conventional EUS transducers has scope for improvement because of the ordinary piezoelectric parameters of Pb(Zr, Ti) [Formula: see text] (PZT) bulk ceramic as well as its inferior mechanical flexibility which can cause material cracks during the circular shaping process. To achieve both prominent imaging capabilities and high device reliability, a 128-element 6.8-MHz circular array transducer is developed using a Pb(Mg [Formula: see text]Nb [Formula: see text]) [Formula: see text]-PbTiO3 (PMN-PT) 1-3 composite with a coefficient of high electromechanical coupling ( [Formula: see text]) and good mechanical flexibility. The characterization results exhibit a large average bandwidth of 58%, a high average sensitivity of 100 mVpp, and a crosstalk of less than -37 dB near the center frequency. Imaging performance of the PMN-PT composite-based array transducer is evaluated by a wire phantom, an anechoic cyst phantom, and an ex-vivo swine intestine. This work demonstrates the superior performance of the crucial ultrasonic device based on an advanced PMN-PT composite material and may lead to the development of next-generation biomedical ultrasonic devices for clinical diagnosis and treatment.

Ultrafast Endoscopic Ultrasonography With Circular Array.

Rapid development of ultrafast ultrasound imaging has led to novel medical ultrasound applications, including shear wave elastography and super-resolution vascular imaging. However, these have yet to incorporate endoscopic ultrasonography (EUS) with a circular array, which provides a wider view in the alimentary canal than traditional linear and convex arrays. A coherent diverging wave compounding (CDWC) imaging method was proposed for ultrafast EUS imaging and implemented on a custom circular array. In CDWC, virtual acoustic point sources are allocated and virtually insonified diverging waves from each source are achieved by adjusting all circular array elements' emission time delays. Diverging waves emitted from different virtual sources are coherently compounded, generating synthetic transmit focusing at every location in the image plane. As the field of view of the circular array is centrally symmetric, all virtual sources are equidistantly distributed on a concentric circle of radius r . To achieve the highest frame rate possible with image quality comparable to that obtained with the traditional multi-focus imaging method, the effects of various radii r and virtual source quantities on the compounded image quality were theoretically analyzed and experimentally verified. Simulation, phantom, and ex-vivo experiments were conducted with an 8 MHz, 124-element circular array, with a 5.35 mm radius. When 16 virtual sources were used with r=1.605 mm, image quality comparable to that obtained with the multi-focus approach was achieved at a frame rate of 1000 frames/s. This demonstrates the feasibility of the proposed ultrafast EUS imaging method and promotes further development of multi-functional EUS devices.

Development of Scalable 2D Plane Array for Transcranial Ultrasonic Neuromodulation on Non-Human Primates: An Ex Vivo Study.

Ultrasonic neuromodulation on large animals, like non-human primates, requires the array transducer to have a good steering ability to arbitrarily stimulate various brain locations. Moreover, due to the different sizes of the animal heads, the array is preferred to be conveniently adjustable to different aperture sizes. To meet these requirements, a scalable 2D plane array, which can consist of up to tens of 256-element square modules, has been designed and fabricated in this study. Arrays with large apertures, consisting of 12 and 48 modules (including 3072 and 12288 elements, respectively), have been assembled and tested. The square modules were driven at a center frequency of 1.04 MHz. The transcranial and steering abilities of the modules were verified in a water tank on an ex vivo macaque skull. A focused beam transmitted by single module could pass through the skull with its peak acoustic pressure reduced to 18.1% of that in the free-field, with an acceptable transverse steering range up to 10 mm. Moreover, a much smaller focal spot was obtained by simultaneous transmission using two vertical-placed modules. Compared to the single module, the focal zone axial size was reduced from 29.7 mm to 3.3 mm, and the "stripes" pattern in the focal zone caused by standing waves was eliminated using a difference-frequency transmitting strategy. Such scalable 2D plane array, which can provide a small-size focal spot and has a large steering range and an acceptable transcranial ability, can be useful in research on ultrasonic neuromodulation on non-human primates.

Transcranial Low-Intensity Pulsed Ultrasound Modulates Structural and Functional Synaptic Plasticity in Rat Hippocampus.

Plasticity of synaptic structure and function play an essential role in neuronal development, cognitive functions, and degenerative diseases. Recently, low-intensity pulsed ultrasound (LIPUS) stimulation has been reported as a promising technology for neuromodulation. However, the effect of LIPUS stimulation on the structural and functional synaptic plasticity in rat hippocampus has not yet been addressed. The aim of this study was to investigate whether LIPUS stimulation could affect the dendritic structure, electrophysiological properties, and expression level of glutamate receptors GluN2A, GluN2B, and GluR1 subunits in rat hippocampus. Transcranial LIPUS was delivered to CA1 of the intact hippocampus of rats ( n = 40 ) for 10 days (10 min/day) with the following parameters: fundamental frequency of 0.5 MHz, pulse repetition frequency (PRF) of 500 Hz, peak negative pressure of 0.42 MPa, and Ispta of 360 mW/cm2. The effect of LIPUS on dendritic structure, electrophysiological properties, and the expression of neurotransmitter receptors was measured using Golgi staining, electrophysiological recording, and western blotting, respectively. Golgi staining and electrophysiological recordings showed that LIPUS stimulation significantly increased the density of dendritic spines (0.72 ± 0.17 versus 0.94 ± 0.19 spines/ [Formula: see text], ) and the frequency of spontaneous excitatory postsynaptic current (0.37 ± 0.14 versus 1.77 ± 0.37 Hz, ) of CA1 hippocampal neurons. Furthermore, the western blotting analysis demonstrated a significant increase in the expression level of GluN2A ( ). The results illustrated the effect of LIPUS on the dendritic structure, function, and neurotransmitter receptors, which may provide a powerful tool for treating neurodegenerative diseases.

[Influencing Mechanism and Spatio-temporal Pattern of Stomatal Ozone Flux of Winter Wheat Under Ozone Pollution].

As one of the main atmospheric pollutants over surface layer,researches on the increasing surface ozone concentration and its impact on main crops have become the focus of every government and the public.In this paper,based on the observations in Nanjing using the main local cultivars in China's major winter wheat producing areas,it was expected to obtain the data including ozone concentration,meteorological data and stomatal conductance by continuous observation.Stomatal conductance model was used and parameterized,combined with flux model,we analyzed the characteristics of stomatal flux in winter wheat under ozone pollution.At the same time,the stomatal conductance and stomatal ozone absorption flux of winter wheat were simulated in Jiangsu Province.The main results were as follows:Elevated ozone concentration could reduce stomatal conductance of winter wheat leaf and stomatal conductance decreased with the increase of ozone concentration.According to the observational data through the experiment,based on the boundary line technology,stomatal conductance model was parameterized to simulate stomatal conductance of wheat leaves from environmental factors.Approximately 90%,77% and 83% variation of measured stomatal conductance could be explained by the stomatal conductance model.In the experiment,the total ozone absorption flux in ozone concentration of CK (53.67 nL·L-1),100nL·L-1,150nL·L-1 was 6.42 mmol·m-2,12.27 mmol·m-2,13.90 mmol·m-2 respectively.The ozone concentration gradually increased from early period to late period during the period of winter wheat growth in Jiangsu area.The average stomatal conductance followed the order of the middle stage >the later stage >the early stage.Winter wheat ozone cumulative absorption flux was the highest during the middle stage.

Quantitative Glycoproteomic Analysis Identifies Platelet-Induced Increase of Monocyte Adhesion via the Up-Regulation of Very Late Antigen 5.

Physiological stimuli, such as thrombin, or pathological stimuli, such as lysophosphatidic acid (LPA), activate platelets circulating in blood. Once activated, platelets bind to monocytes via P-selectin-PSGL-1 interactions but also release the stored contents of their granules. These platelet releasates, in addition to direct platelet binding, activate monocytes and facilitate their recruitment to atherosclerotic sites. Consequently, understanding the changes platelet releasates induce in monocyte membrane proteins is critical. We studied the glyco-proteome changes of THP-1 monocytic cells affected by LPA- or thrombin-induced platelet releasates. We employed lectin affinity chromatography combined with filter aided sample preparation to achieve high glyco- and membrane protein and protein sequence coverage. Using stable isotope labeling by amino acids in cell culture, we quantified 1715 proteins, including 852 membrane and 500 glycoproteins, identifying the up-regulation of multiple proteins involved in monocyte extracellular matrix binding and transendothelial migration. Flow cytometry indicated expression changes of integrin α5, integrin ß1, PECAM-1, and PSGL-1. The observed increase in monocyte adhesion to fibronectin was determined to be mediated by the up-regulation of very late antigen 5 via a P-selectin-PSGL-1 independent mechanism. This novel aspect could be validated on CD14+ human primary monocytes, highlighting the benefits of the improved enrichment method regarding high membrane protein coverage and reliable quantification.

Identification of total reversible cysteine oxidation in an atherosclerosis model using a modified biotin switch assay.

Oxidative stress due to the imbalance of reactive oxygen species (ROS) and the resulting reversible cysteine oxidation (CysOX) are involved in the early proatherogenic aspect of atherosclerosis. Given that the corresponding redox signaling pathways are still unclear, a modified biotin switch assay was developed to quantify the reversible CysOX in an atherosclerosis model established by using a monocytic cell line treated with platelet releasate. The accumulation of ROS was observed in the model system and validated in human primary monocytes. Through the application of the modified biotin switch assay, we obtained the first reversible CysOX proteome for this model. A total of 75 peptides, corresponding to 53 proteins, were quantified with oxidative modification. The bioinformatics analysis of these CysOX-containing proteins highlighted biological processes including glycolysis, cytoskeleton arrangement, and redox regulation. Moreover, the reversible oxidation of three glycolysis enzymes was observed using this method, and the regulation influence was verified by an enzyme activity assay. NADPH oxidase (NOX) inhibition treatment, in conjunction with the modified biotin switch method, was used to evaluate the global CysOX status. In conclusion, this versatile modified biotin switch assay provides an approach for the quantification of all reversible CysOX and for the study of redox signaling in atherosclerosis as well as in diseases in other biological systems.