Ascorbyl palmitate ameliorates inflammatory diseases by inhibition of NLRP3 inflammasome.

The aberrant activation of NLRP3 inflammasome contributes to pathogenesis of multiple inflammation-driven human diseases. However, the medications targeting NLRP3 inflammasome are not approved for clinic use to date. Here, we show that ascorbyl palmitate (AP), a lipophilic derivative of ascorbic acid (AA) and a safe food additive, is a potent inhibitor of NLRP3 inflammasome. Compared with AA, AP inhibited the activation of NLRP3 inflammasome with increased potency and specificity. Mechanistically, AP directly scavenged mitochondrial reactive oxygen species (mitoROS) by its antioxidant activity and blocked NLRP3-NEK7 interaction and NLRP3 inflammasome assembly. Moreover, AP showed more significant preventive effects than AA in LPS-induced systemic inflammation, dextran sulfate sodium (DSS)-induced colitis and experimental autoimmune encephalomyelitis (EAE). Thus, our results suggest that AP is a potential therapeutic combating NLRP3-driven diseases.

Immunophilin FKB20-2 participates in oligomerization of Photosystem I in Chlamydomonas.

PSI is a sophisticated photosynthesis protein complex that fuels the light reaction of photosynthesis in algae and vascular plants. While the structure and function of PSI have been studied extensively, the dynamic regulation on PSI oligomerization and high light response is less understood. In this work, we characterized a high light-responsive immunophilin gene FKB20-2 (FK506-binding protein 20-2) required for PSI oligomerization and high light tolerance in Chlamydomonas (Chlamydomonas reinhardtii). Biochemical assays and 77-K fluorescence measurement showed that loss of FKB20-2 led to the reduced accumulation of PSI core subunits and abnormal oligomerization of PSI complexes and, particularly, reduced PSI intermediate complexes in fkb20-2. It is noteworthy that the abnormal PSI oligomerization was observed in fkb20-2 even under dark and dim light growth conditions. Coimmunoprecipitation, MS, and yeast 2-hybrid assay revealed that FKB20-2 directly interacted with the low molecular weight PSI subunit PsaG, which might be involved in the dynamic regulation of PSI-light-harvesting complex I supercomplexes. Moreover, abnormal PSI oligomerization caused accelerated photodamage to PSII in fkb20-2 under high light stress. Together, we demonstrated that immunophilin FKB20-2 affects PSI oligomerization probably by interacting with PsaG and plays pivotal roles during Chlamydomonas tolerance to high light.

SWENet: A Physics-Informed Deep Neural Network (PINN) for Shear Wave Elastography.

Shear wave elastography (SWE) enables the measurement of elastic properties of soft materials in a non-invasive manner and finds broad applications in various disciplines. The state-of-the-art SWE methods rely on the measurement of local shear wave speeds to infer material parameters and suffer from wave diffraction when applied to soft materials with strong heterogeneity. In the present study, we overcome this challenge by proposing a physics-informed neural network (PINN)-based SWE (SWENet) method. The spatial variation of elastic properties of inhomogeneous materials has been introduced in the governing equations, which are encoded in SWENet as loss functions. Snapshots of wave motions have been used to train neural networks, and during this course, the elastic properties within a region of interest illuminated by shear waves are inferred simultaneously. We performed finite element simulations, tissue-mimicking phantom experiments, and ex vivo experiments to validate the method. Our results show that the shear moduli of soft composites consisting of matrix and inclusions of several millimeters in cross-section dimensions with either regular or irregular geometries can be identified with excellent accuracy. The advantages of the SWENet over conventional SWE methods consist of using more features of the wave motions and enabling seamless integration of multi-source data in the inverse analysis. Given the advantages of SWENet, it may find broad applications where full wave fields get involved to infer heterogeneous mechanical properties, such as identifying small solid tumors with ultrasound SWE, and differentiating gray and white matters of the brain with magnetic resonance elastography.

Simultaneous tensile and shear measurement of the human cornea in vivo using S0- and A0-wave optical coherence elastography.

Understanding corneal stiffness is valuable for improving refractive surgery, detecting corneal abnormalities, and assessing intraocular pressure. However, accurately measuring the elastic properties, specifically the tensile and shear moduli that govern mechanical deformation, has been challenging. To tackle this issue, we have developed guided-wave optical coherence elastography that can simultaneously excite and analyze symmetric (S0) and anti-symmetric (A0) elastic waves in the cornea at around 10 kHz frequencies, enabling us to extract tensile and shear properties from measured wave dispersion curves. We verified the technique using elastomer phantoms and ex vivo porcine corneas and investigated the dependence on intraocular pressure using acoustoelastic theory that incorporates corneal tension and a nonlinear constitutive tissue model. In a pilot study involving six healthy human subjects aged 31 to 62, we measured shear moduli (Gzx) of 94±20 kPa (mean±standard deviation) and tensile moduli (Exx) of 4.0±1.1 MPa at central corneas. Our preliminary analysis of age-dependence revealed contrasting trends: -8.3±4.5 kPa/decade for shear and 0.30±0.21 MPa/decade for tensile modulus. This OCE technique has the potential to become a highly useful clinical tool for the quantitative biomechanical assessment of the cornea. STATEMENT OF SIGNIFICANCE: This article reports an innovative elastography technique using two guided elastic waves, demonstrating the measurement of both tensile and shear moduli in human cornea in vivo with unprecedented precision. This technique paves the way for comprehensive investigations into corneal mechanics and holds clinical significance in various aspects of corneal health and disease management.

Vacuolar Sugar Transporter TMT2 Plays Crucial Roles in Germination and Seedling Development in Arabidopsis.

Vacuolar sugar transporters transport sugar across the tonoplast, are major players in maintaining sugar homeostasis, and therefore play vital roles in plant growth, development, and biomass yield. In this study, we analyzed the physiological roles of the tonoplast monosaccharide transporter 2 (TMT2) in Arabidopsis. In contrast to the wild type (WT) that produced uniform seedlings, the tmt2 mutant produced three types of offspring: un-germinated seeds (UnG), seedlings that cannot form true leaves (tmt2-S), and seedlings that develop normally (tmt2-L). Sucrose, glucose, and fructose can substantially, but not completely, rescue the abnormal phenotypes of the tmt2 mutant. Abnormal cotyledon development, arrested true leaf development, and abnormal development of shoot apical meristem (SAM) were observed in tmt2-S seedlings. Cotyledons from the WT and tmt2-L seedlings restored the growth of tmt2-S seedlings through micrografting. Moreover, exogenous sugar sustained normal growth of tmt2-S seedlings with cotyledon removed. Finally, we found that the TMT2 deficiency resulted in growth defects, most likely via changing auxin signaling, target of rapamycin (TOR) pathways, and cellular nutrients. This study unveiled the essential functions of TMT2 for seed germination and initial seedling development, ensuring cotyledon function and mobilizing sugars from cotyledons to seedlings. It also expanded the current knowledge on sugar metabolism and signaling. These findings have fundamental implications for enhancing plant biomass production or seed yield in future agriculture.

In vivo Optical Coherence Elastography Unveils Spatial Variation of Human Corneal Stiffness.

OBJECTIVE: The mechanical properties of corneal tissues play a crucial role in determining corneal shape and have significant implications in vision care. This study aimed to address the challenge of obtaining accurate in vivo data for the human cornea. METHODS: We have developed a high-frequency optical coherence elastography (OCE) technique using shear-like antisymmetric (A0)-mode Lamb waves at frequencies above 10 kHz. RESULTS: By incorporating an anisotropic, nonlinear constitutive model and utilizing the acoustoelastic theory, we gained quantitative insights into the influence of corneal tension on wave speeds and elastic moduli. Our study revealed significant spatial variations in the shear modulus of the corneal stroma on healthy subjects for the first time. Over an age span from 21 to 34 (N = 6), the central corneas exhibited a mean shear modulus of 87 kPa, while the corneal periphery showed a significant decrease to 44 kPa. The central cornea's shear modulus decreases with age with a slope of -19 +/- 8 kPa per decade, whereas the periphery showed non-significant age dependence. The limbus demonstrated an increased shear modulus exceeding 100 kPa. We obtained wave displacement profiles that are consistent with highly anisotropic corneal tissues. CONCLUSION: Our approach enabled precise measurement of corneal tissue elastic moduli in situ with high precision (< 7%) and high spatial resolution (< 1 mm). Our results revealed significant stiffness variation from the central to peripheral corneas. SIGNIFICANCE: The high-frequency OCE technique holds promise for biomechanical evaluation in clinical settings, providing valuable information for refractive surgeries, degenerative disorder diagnoses, and intraocular pressure assessments.

Poster Session: Optical coherence elastography for In situ measurement of stiffness increase in posterior sclera after crosslinking.

During eye growth, scleral development critically determine eye size and thus the refractive status of the eye. Scleral remodeling in myopia includes scleral thinning, loss of scleral tissue, and weakening of the mechanical properties. Therefore, an intervention aiming at stiffening scleral tissues (crosslinking, SCXL) may provide a way to prevent or treat myopia. The development of SCXL requires tools to evaluate the effects of crosslinking on the mechanical properties of tissues, particularly in sclera where the mechanical properties are more spatially heterogeneous than in the cornea, anisotropic, and varying locally from the anterior to posterior regions. Here, we apply the high-frequency OCE technique to measure the heterogeneous mechanical properties of posterior scleral tissues and, evaluate the changes in shear moduli after SCXL. As a model system, we use ex vivo in porcine eyes and riboflavin-assisted UV crosslinking. From measured elastic wave speeds (6-16kHz), the average out-of-plane shear modulus was 0.71±0.12MPa (n=20) for normal scleras. After treatment, the shear modulus increased to 1.50±0.39MPa. This 2-fold change was consistent with the increase of static Young's modulus from 5.5±.1 to 9.3±1.9MPa after crosslinking, using conventional uniaxial extensometry. OCE revealed regional stiffness differences across the temporal, nasal, and deeper posterior sclera, demonstrating its potential as a noninvasive tool to test the effect of scleral crosslinking.

Simultaneous tensile and shear measurement of the human cornea in vivo using S0- and A0-wave optical coherence elastography.

Understanding corneal stiffness is valuable for improving refractive surgery, detecting corneal abnormalities, and assessing intraocular pressure. However, accurately measuring the elastic properties, particularly the tensile and shear moduli that govern mechanical deformation, has been challenging. To tackle this issue, we have developed guided-wave optical coherence elastography that can simultaneously excite and analyze symmetric (S0) and anti-symmetric (A0) elastic waves in the cornea at frequencies around 10 kHz and allows us to extract tensile and shear properties from measured wave dispersion curves. By applying acoustoelastic theory that incorporates corneal tension and a nonlinear constitutive tissue model, we verified the technique using elastomer phantoms and ex vivo porcine corneas and investigated the dependence on intraocular pressure. For two healthy human subjects, we measured a mean tensile modulus of 3.6 MPa and a mean shear modulus of 76 kPa in vivo with estimated errors of < 4%. This technique shows promise for the quantitative biomechanical assessment of the cornea in a clinical setting.

Ultra-wideband optical coherence elastography from acoustic to ultrasonic frequencies.

Visualizing viscoelastic waves in materials and tissues through noninvasive imaging is valuable for analyzing their mechanical properties and detecting internal anomalies. However, traditional elastography techniques have been limited by a maximum wave frequency below 1-10 kHz, which hampers temporal and spatial resolution. Here, we introduce an optical coherence elastography technique that overcomes the limitation by extending the frequency range to MHz. Our system can measure the stiffness of hard materials including bones and extract viscoelastic shear moduli for polymers and hydrogels in conventionally inaccessible ranges between 100 Hz and 1 MHz. The dispersion of Rayleigh surface waves across the ultrawide band allowed us to profile depth-dependent shear modulus in cartilages ex vivo and human skin in vivo with sub-mm anatomical resolution. This technique holds immense potential as a noninvasive measurement tool for material sciences, tissue engineering, and medical diagnostics.

Noninvasive measurement of local stress inside soft materials with programmed shear waves.

Mechanical stresses across different length scales play a fundamental role in understanding biological systems' functions and engineering soft machines and devices. However, it is challenging to noninvasively probe local mechanical stresses in situ, particularly when the mechanical properties are unknown. We propose an acoustoelastic imaging-based method to infer the local stresses in soft materials by measuring the speeds of shear waves induced by custom-programmed acoustic radiation force. Using an ultrasound transducer to excite and track the shear waves remotely, we demonstrate the application of the method by imaging uniaxial and bending stresses in an isotropic hydrogel and the passive uniaxial stress in a skeletal muscle. These measurements were all done without the knowledge of the constitutive parameters of the materials. The experiments indicate that our method will find broad applications, ranging from health monitoring of soft structures and machines to diagnosing diseases that alter stresses in soft tissues.

In situ measurement of the stiffness increase in the posterior sclera after UV-riboflavin crosslinking by optical coherence elastography.

Scleral crosslinking may provide a way to prevent or treat myopia by stiffening scleral tissues. The ability to measure the stiffness of scleral tissues in situ pre and post scleral crosslinking would be useful but has not been established. Here, we tested the feasibility of optical coherence elastography (OCE) to measure shear modulus of scleral tissues and evaluate the impact of crosslinking on different posterior scleral regions using ex vivo porcine eyes as a model. From measured elastic wave speeds at 6 - 16 kHz, we obtained out-of-plane shear modulus value of 0.71 ± 0.12 MPa (n = 20) for normal porcine scleral tissues. After riboflavin-assisted UV crosslinking, the shear modulus increased to 1.50 ± 0.39 MPa (n = 20). This 2-fold change was consistent with the increase of static Young's modulus from 5.5 ± 1.1 MPa to 9.3 ± 1.9 MPa after crosslinking, which we measured using conventional uniaxial extensometry on tissue stripes. OCE revealed regional stiffness differences across the temporal, nasal, and deeper posterior sclera. Our results show the potential of OCE as a noninvasive tool to evaluate the effect of scleral crosslinking.

In vivo stiffness measurement of epidermis, dermis, and hypodermis using broadband Rayleigh-wave optical coherence elastography.

Traveling-wave optical coherence elastography (OCE) is a promising technique to measure the stiffness of biological tissues. While OCE has been applied to relatively homogeneous samples, tissues with significantly varying elasticity through depth pose a challenge, requiring depth-resolved measurement with sufficient resolution and accuracy. Here, we develop a broadband Rayleigh-wave OCE technique capable of measuring the elastic moduli of the 3 major skin layers (epidermis, dermis, and hypodermis) reliably by analyzing the dispersion of leaky Rayleigh surface waves over a wide frequency range of 0.1-10 kHz. We show that a previously unexplored, high frequency range of 4-10 kHz is critical to resolve the thin epidermis, while a low frequency range of 0.2-1 kHz is adequate to probe the dermis and deeper hypodermis. We develop a dual bilayer-based inverse model to determine the elastic moduli in all 3 layers and verify its high accuracy with finite element analysis and skin-mimicking phantoms. Finally, the technique is applied to measure the forearm skin of healthy volunteers. The Young's modulus of the epidermis (including the stratum corneum) is measured to be ∼ 4 MPa at 4-10 kHz, whereas Young's moduli of the dermis and hypodermis are about 40 and 15 kPa, respectively, at 0.2-1 kHz. Besides dermatologic applications, this method may be useful for the mechanical analysis of various other layered tissues with sub-mm depth resolution. STATEMENT OF SIGNIFICANCE: To our knowledge, this is the first study that resolves the stiffness of the thin epidermis from the dermis and hypodermis, made possible by using high-frequency (4 - 10 kHz) elastic waves and optical coherence elastography. Beyond the skin, this technique may be useful for mechanical characterizations of various layered biomaterials and tissues.

Probing the Mechanical Properties of Large Arteries by Measuring Their Deformation In Vivo with Ultrasound.

Aging and cardiovascular diseases (CVDs) may alter the microstructures of arteries and hence their mechanical properties. Therefore, the measurement of intrinsic artery mechanical properties in vivo can provide valuable information in understanding aging and CVDs and is of clinical significance. The accuracy of advanced ultrasound imaging techniques in measuring the deformation of large arteries under blood pressure is good. However, the assessment of arterial stiffness in vivo remains a challenge. An inverse method to infer the constitutive parameters of arteries in vivo from the blood pressure-arterial radius relationship (P-r curve) is proposed here. The stability analysis reveals that a key constitutive parameter, bθ, which measures the circumferential hardening of an artery, can be reliably identified. An in vivo experiment was performed on the common carotid arteries of 41 healthy volunteers (age: 37 ± 17 y). The value of bθ varies significantly (from 0.55 ± 0.15 for the young group to 0.93 ± 0.29 for the older group, p < 0.01) and is positively correlated with age (r = 0.673, p < 0.01). Furthermore, our theoretical analysis and experimental study have revealed a strong correlation between the clinic-used stiffness index ß and bθ. This study shows that the arterial material parameter bθ can be measured in vivo, which makes it promising as a new biomarker in the diagnosis of CVDs.

Arterial Stiffness Probed by Dynamic Ultrasound Elastography Characterizes Waveform of Blood Pressure.

The clinical and economic burdens of cardiovascular diseases pose a global challenge. Growing evidence suggests an early assessment of arterial stiffness can provide insights into the pathogenesis of cardiovascular diseases. However, it remains difficult to quantitatively characterize local arterial stiffness in vivo. Here we utilize guided axial waves continuously excited and detected by ultrasound to probe local blood pressures and mechanical properties of common carotid arteries simultaneously. In a pilot study of 17 healthy volunteers, we observe a  âˆ¼ 20 % variation in the group velocities of the guided axial waves (5.16 ± 0.55 m/s in systole and 4.31 ± 0.49 m/s in diastole) induced by the variation of the blood pressures. A linear relationship between the square of group velocity and blood pressure is revealed by the experiments and finite element analysis, which enables us to measure the waveform of the blood pressures by the group velocities. Furthermore, we propose a wavelet analysis-based method to extract the dispersion relations of the guided axial waves. We then determined the shear modulus by fitting the dispersion relations in diastole with the leaky Lamb wave model. The average shear modulus of all the volunteers is 166.3 ± 32.8 kPa. No gender differences are found. This study shows the group velocity and dispersion relation of the guided axial waves can be utilized to probe blood pressure and arterial stiffness locally in a noninvasive manner and thus promising for early diagnosis of cardiovascular diseases.

Non-destructive mapping of stress and strain in soft thin films through sound waves.

Measuring the in-plane mechanical stress in a taut membrane is challenging, especially if its material parameters are unknown or altered by the stress. Yet being able to measure the stress is of fundamental interest to basic research and practical applications that use soft membranes, from engineering to tissues. Here we present a robust non-destructive technique to measure directly in-situ stress and strain in soft thin films without the need to calibrate material parameters. Our method relies on measuring the speed of elastic waves propagating in the film. Using optical coherence tomography, we verify our method experimentally for a stretched rubber membrane, a piece of cling film (about 10 µm thick), and the leather skin of a traditional Irish frame drum. We find that our stress predictions are highly accurate and anticipate that our technique could be useful in applications ranging from soft matter devices to biomaterial engineering and medical diagnosis.

Supershear surface waves reveal prestress and anisotropy of soft materials.

Surface waves play important roles in many fundamental and applied areas from seismic detection to material characterizations. Supershear surface waves with propagation speeds greater than bulk shear waves have recently been reported, but their properties are not well understood. Here we describe theoretical and experimental results on supershear surface waves in rubbery materials. We find that supershear surface waves can be supported in viscoelastic materials with no restriction on the shear quality factor. Interestingly, the effect of prestress on the speed of the supershear surface wave is opposite to that of the Rayleigh surface wave. Furthermore, anisotropy of material affects the supershear wave much more strongly than the Rayleigh surface wave. We offer heuristic interpretation as well as theoretical verification of our experimental observations. Our work points to the potential applications of supershear waves for characterizing the bulk mechanical properties of soft solid from the free surface.

Inhibition of the Inflammasome Activity of NLRP3 Attenuates HDM-Induced Allergic Asthma.

Inhaled allergens promote inflammatory response, tissue damage, and airway hyperresponsiveness in the lungs, leading to allergic asthma. NLRP3, as an immune sensor of infections and cellular stress, is associated with the development and exacerbation of asthma. However, the mechanism by which NLRP3 affects asthma requires further investigation. Here, we showed that inhaled house dust mite (HDM) promotes NLRP3 inflammasome activation in the lungs and specifically induces the maturation of caspase-1 and IL-1ß in alveolar macrophages (AMs). Using Nlrp3-mutant mice, we found that NLRP3 promotes the inflammatory response and pathogenesis in HDM-induced allergic asthma in an inflammasome-dependent manner. Treatment with RRx-001, an NLRP3 inhibitor, significantly reduced inflammatory cell infiltration and mucus secretion in the airway. Our results showed that NLRP3 in myeloid cells promoted the development and progression of allergic asthma in an inflammasome-dependent manner. Small molecules targeting the NLRP3 inflammasome may provide new treatment options for this disease.

Sinapic Acid Reduces Oxidative Stress and Pyroptosis via Inhibition of BRD4 in Alcoholic Liver Disease.

Alcoholic liver disease (ALD) is one of the main causes of death in chronic liver disease. Oxidative stress and pyroptosis are important factors leading to ALD. Bromodomain-containing protein 4 (BRD4) is a factor that we have confirmed to regulate ALD. As a phenolic acid compound, sinapic acid (SA) has significant effects in antioxidant, anti-inflammatory and liver protection. In this study, we explored whether SA regulates oxidative stress and pyroptosis through BRD4 to play a protective effect in ALD. Male C57BL/6 mice and AML-12 cells were used for experiments. We found that SA treatment largely abolished the up-regulation of BRD4 and key proteins of the canonical pyroptosis signalling in the liver of mice fed with alcohol, while conversely enhanced the antioxidant response. Consistantly, both SA pretreatment and BRD4 knockdown inhibited oxidative stress, pyroptosis, and liver cell damage in vitro. More importantly, the expression levels of BRD4 and pyroptosis indicators increased significantly in ALD patients. Molecule docking analysis revealed a potent binding of SA with BRD4. In conclusion, this study demonstrates that SA reduces ALD through BRD4, which is a valuable lead compound that prevents the ALD process.

High-contrast OPCPA front end in high-power petawatt laser facility based on the ps-OPCPA seed system.

A high-energy, high-beam-quality, high-contrast picosecond optical parametric chirped-pulse amplification (ps-OPCPA) laser system was demonstrated. The pulse from a femtosecond oscillator was stretched to 4 ps, after which it was amplified from 140 pJ to 600 µJ by an 8 ps/6 mJ pump laser in two non-collinear OPCPA stages. The total gain was >106, and the root mean square of the energy stability of the laser system was 1.6% in 10 h. The contrasts of the solid and fiber mode-locked femtosecond oscillator-seeded ps-OPCPA systems were compared, and a signal-to-noise ratio of >1011 was achieved. Using this system, the contrast of the front end in high-power picosecond petawatt laser facility was improved by ∼40 dB to >1011, beyond ∼200 ps ahead of the main pulse with an output level of 60 mJ.

AFM-based indentation method for measuring the relaxation property of living cells.

Probing the mechanical properties of cells is critical for understanding their deformation behaviors and biological functions. Although some methods have been proposed to characterize the elastic properties of cells, it is still difficult to measure their time-dependent properties. This paper investigates the use of atomic force microscope (AFM) to determine the reduced relaxation modulus of cells. In principle, AFM is hard to perform an indentation relaxation test that requires a constant indenter displacement during load relaxation, whereas the real AFM indenter displacement usually varies with time during relaxation due to the relatively small bending stiffness of its cantilever. We investigate this issue through a combined theoretical, computational, and experimental effort. A protocol relying on the choice of appropriate cantilever bending stiffness is proposed to perform an AFM-based indentation relaxation test of cells, which enables the measurement of reduced relaxation modulus with high accuracy. This protocol is first validated by performing nanoindentation relaxation tests on a soft material and by comparing the results with those from independent measurements. Then indentation tests of cartilage cells are conducted to demonstrate this method in determining time-dependent properties of living cells. Finally, the change in the viscoelasticity of MCF-7 cells under hyperthermia is investigated.