Pyroptosis of Vascular Smooth Muscle Cells as a Potential New Target for Preventing Vascular Diseases.

Vascular remodeling is the adaptive response of the vessel wall to physiological and pathophysiological changes, closely linked to vascular diseases. Vascular smooth muscle cells (VSMCs) play a crucial role in this process. Pyroptosis, a form of programmed cell death characterized by excessive release of inflammatory factors, can cause phenotypic transformation of VSMCs, leading to their proliferation, migration, and calcification-all of which accelerate vascular remodeling. Inhibition of VSMC pyroptosis can delay this process. This review summarizes the impact of pyroptosis on VSMCs and the pathogenic role of VSMC pyroptosis in vascular remodeling. We also discuss inhibitors of key proteins in pyroptosis pathways and their effects on VSMC pyroptosis. These findings enhance our understanding of the pathogenesis of vascular remodeling and provide a foundation for the development of novel medications that target the control of VSMC pyroptosis as a potential treatment strategy for vascular diseases.

Diffusiophoretic Particle Penetration into Bacterial Biofilms.

Bacterial biofilms are communities of cells adhered to surfaces. These communities represent a predominant form of bacterial life on Earth. A defining feature of a biofilm is the three-dimensional extracellular polymer matrix that protects resident cells by acting as a mechanical barrier to the penetration of chemicals, such as antimicrobials. Beyond being recalcitrant to antibiotic treatment, biofilms are notoriously difficult to remove from surfaces. A promising, but relatively underexplored, approach to biofilm control is to disrupt the extracellular polymer matrix by enabling penetration of particles to increase the susceptibility of biofilms to antimicrobials. In this work, we investigate externally imposed chemical gradients as a mechanism to transport polystyrene particles into bacterial biofilms. We show that preconditioning the biofilm with a prewash step using deionized (DI) water is essential for altering the biofilm so it takes up the micro- and nanoparticles by the application of a further chemical gradient created by an electrolyte. Using different particles and chemicals, we document the transport behavior that leads to particle motion into the biofilm and its further reversal out of the biofilm. Our results demonstrate the importance of chemical gradients in disrupting the biofilm matrix and regulating particle transport in crowded macromolecular environments, and suggest potential applications of particle transport and delivery in other physiological systems.

Confinement, chaotic transport, and trapping of active swimmers in time-periodic flows.

Microorganisms encounter complex unsteady flows, including algal blooms in marine settings, microbial infections in airways, and bioreactors for vaccine and biofuel production. Here, we study the transport of active swimmers in two-dimensional time-periodic flows using Langevin simulations and experiments with swimming bacteria. We find that long-term swimmer transport is controlled by two parameters, the pathlength of the unsteady flow and the normalized swimmer speed. The pathlength nonmonotonically controls swimmer dispersion dynamics, giving rise to three distinct dispersion regimes. Weak flows hinder swimmer transport by confining cells toward flow manifolds. As pathlength increases, chaotic transport along flow manifolds initiates, maximizing the number of unique flow cells traveled. Last, strong flows trap swimmers at the vortex core, suppressing dispersal. Experiments with Vibrio cholerae showed qualitative agreement with model dispersion patterns. Our results reveal that nontrivial chaotic transport can arise in simple unsteady flows and suggest a potentially optimal dispersal strategy for microswimmers in nature.

Quorum-sensing control of matrix protein production drives fractal wrinkling and interfacial localization of Vibrio cholerae pellicles.

Bacterial cells at fluid interfaces can self-assemble into collective communities with stunning macroscopic morphologies. Within these soft, living materials, called pellicles, constituent cells gain group-level survival advantages including increased antibiotic resistance. However, the regulatory and structural components that drive pellicle self-patterning are not well defined. Here, using Vibrio cholerae as our model system, we report that two sets of matrix proteins and a key quorum-sensing regulator jointly orchestrate the sequential mechanical instabilities underlying pellicle morphogenesis, culminating in fractal patterning. A pair of matrix proteins, RbmC and Bap1, maintain pellicle localization at the interface and prevent self-peeling. A single matrix protein, RbmA, drives a morphogenesis program marked by a cascade of ever finer wrinkles with fractal scaling in wavelength. Artificial expression of rbmA restores fractal wrinkling to a ΔrbmA mutant and enables precise tuning of fractal dimensions. The quorum-sensing regulatory small RNAs Qrr1-4 first activate matrix synthesis to launch pellicle primary wrinkling and ridge instabilities. Subsequently, via a distinct mechanism, Qrr1-4 suppress fractal wrinkling to promote fine modulation of pellicle morphology. Our results connect cell-cell signaling and architectural components to morphogenic patterning and suggest that manipulation of quorum-sensing regulators or synthetic control of rbmA expression could underpin strategies to engineer soft biomaterial morphologies on demand.

Quorum-sensing- and type VI secretion-mediated spatiotemporal cell death drives genetic diversity in Vibrio cholerae.

Bacterial colonies composed of genetically identical individuals can diversify to yield variant cells with distinct genotypes. Variant outgrowth manifests as sectors. Here, we show that Type VI secretion system (T6SS)-driven cell death in Vibrio cholerae colonies imposes a selective pressure for the emergence of variant strains that can evade T6SS-mediated killing. T6SS-mediated cell death occurs in two distinct spatiotemporal phases, and each phase is driven by a particular T6SS toxin. The first phase is regulated by quorum sensing and drives sectoring. The second phase does not require the T6SS-injection machinery. Variant V. cholerae strains isolated from colony sectors encode mutated quorum-sensing components that confer growth advantages by suppressing T6SS-killing activity while simultaneously boosting T6SS-killing defenses. Our findings show that the T6SS can eliminate sibling cells, suggesting a role in intra-specific antagonism. We propose that quorum-sensing-controlled T6SS-driven killing promotes V. cholerae genetic diversity, including in natural habitats and during disease.

Bacteria hinder large-scale transport and enhance small-scale mixing in time-periodic flows.

Understanding mixing and transport of passive scalars in active fluids is important to many natural (e.g., algal blooms) and industrial (e.g., biofuel, vaccine production) processes. Here, we study the mixing of a passive scalar (dye) in dilute suspensions of swimming Escherichia coli in experiments using a two-dimensional (2D) time-periodic flow and in a simple simulation. Results show that the presence of bacteria hinders large-scale transport and reduces overall mixing rate. Stretching fields, calculated from experimentally measured velocity fields, show that bacterial activity attenuates fluid stretching and lowers flow chaoticity. Simulations suggest that this attenuation may be attributed to a transient accumulation of bacteria along regions of high stretching. Spatial power spectra and correlation functions of dye-concentration fields show that the transport of scalar variance across scales is also hindered by bacterial activity, resulting in an increase in average size and lifetime of structures. On the other hand, at small scales, activity seems to enhance local mixing. One piece of evidence is that the probability distribution of the spatial concentration gradients is nearly symmetric with a vanishing skewness. Overall, our results show that the coupling between activity and flow can lead to nontrivial effects on mixing and transport.

Hierarchical transitions and fractal wrinkling drive bacterial pellicle morphogenesis.

Bacterial cells can self-organize into structured communities at fluid-fluid interfaces. These soft, living materials composed of cells and extracellular matrix are called pellicles. Cells residing in pellicles garner group-level survival advantages such as increased antibiotic resistance. The dynamics of pellicle formation and, more generally, how complex morphologies arise from active biomaterials confined at interfaces are not well understood. Here, using Vibrio cholerae as our model organism, a custom-built adaptive stereo microscope, fluorescence imaging, mechanical theory, and simulations, we report a fractal wrinkling morphogenesis program that differs radically from the well-known coalescence of wrinkles into folds that occurs in passive thin films at fluid-fluid interfaces. Four stages occur: growth of founding colonies, onset of primary wrinkles, development of secondary curved ridge instabilities, and finally the emergence of a cascade of finer structures with fractal-like scaling in wavelength. The time evolution of pellicle formation depends on the initial heterogeneity of the film microstructure. Changing the starting bacterial seeding density produces three variations in the sequence of morphogenic stages, which we term the bypass, crystalline, and incomplete modes. Despite these global architectural transitions, individual microcolonies remain spatially segregated, and thus, the community maintains spatial and genetic heterogeneity. Our results suggest that the memory of the original microstructure is critical in setting the morphogenic dynamics of a pellicle as an active biomaterial.

Cell position fates and collective fountain flow in bacterial biofilms revealed by light-sheet microscopy.

Bacterial biofilms represent a basic form of multicellular organization that confers survival advantages to constituent cells. The sequential stages of cell ordering during biofilm development have been studied in the pathogen and model biofilm-former Vibrio cholerae It is unknown how spatial trajectories of individual cells and the collective motions of many cells drive biofilm expansion. We developed dual-view light-sheet microscopy to investigate the dynamics of biofilm development from a founder cell to a mature three-dimensional community. Tracking of individual cells revealed two distinct fates: one set of biofilm cells expanded ballistically outward, while the other became trapped at the substrate. A collective fountain-like flow transported cells to the biofilm front, bypassing members trapped at the substrate and facilitating lateral biofilm expansion. This collective flow pattern was quantitatively captured by a continuum model of biofilm growth against substrate friction. Coordinated cell movement required the matrix protein RbmA, without which cells expanded erratically. Thus, tracking cell lineages and trajectories in space and time revealed how multicellular structures form from a single founder cell.

Three-dimensional structures and symmetry breaking in viscoelastic cross-channel flow.

Using holographic particle tracking, we report the three-dimensional flow structure organizing the viscoelastic instability in cross-channel flow. Beyond a critical Wi, the advective core flow undergoes an out-of-plane instability marked by the emergence of tertiary flow, resembling that of the toroidal vortices in Taylor-Couette geometry. The out-of-plane flow component distorts the separatrix between the impinging inflow streams, triggering symmetry breaking normal to the extension plane. As extensional rate increases, progressively higher order modes of the separatrix are observed, akin to Euler buckling of a rigid column. The disturbances propagate upstream via stress fluctuations despite viscous dissipation. These complex flow structures may be generic to elastic turbulence in mixed flows.

Flow Resistance and Structures in Viscoelastic Channel Flows at Low Re.

The flow of viscoelastic fluids in channels and pipes remains poorly understood, particularly at low Reynolds numbers. Here, we investigate the flow of polymeric solutions in straight channels using pressure measurements and particle tracking. The flow friction factor f_{η} versus flow rate exhibits two regimes: a transitional regime marked by rapid increase in drag, and a turbulentlike regime characterized by a sudden decrease in drag and a weak dependence on flow rate. Lagrangian trajectories show finite transverse modulations not seen in Newtonian fluids. These curvature perturbations far downstream can generate sufficient hoop stresses to sustain the flow instabilities in the parallel shear flow.

Upstream vortex and elastic wave in the viscoelastic flow around a confined cylinder.

The viscoelastic flow past a cylinder is a classic benchmark problem that is not completely understood. Using novel 3D holographic particle velocimetry, we report three main discoveries of the elastic instability upstream of a single cylinder in viscoelastic channel flow. First, we observe that upstream vortices initiate at the corner between the cylinder and the wall and grow with increasing flow rate. Second, beyond a critical Weissenberg, the flow upstream becomes unsteady and switches between two bistable configurations, leading to symmetry breaking in the cylinder axis direction that is highly three-dimensional in nature. Lastly, we find that the disturbance of the elastic instability propagates relatively far upstream via an elastic wave, and is weakly correlated with that in the cylinder wake. The wave speed and the extent of the instability increase with Weissenberg number, indicating an absolute instability in viscoelastic fluids.

In vitro and in vivo metabolic activation of rhein and characterization of glutathione conjugates derived from rhein.

Rhein (RH), 4,5-dihydroxyanthrauinone-2-carboxylic acid, is found in rhubarb (Dahuang), a traditional herbal medicine. RH has reportedly demonstrated multiple pharmacologic properties. Previous studies have also shown that RH induced hepatotoxicity, but the mechanisms of the adverse effect remain unknown. The major objective of the present study was to study the metabolic pathways of RH in order to identify potential reactive metabolites. One mono-hydroxylation metabolite (M1) was detected in urine and bile of rats given RH. M1 was also observed in rat and human liver microsomal incubations after exposure to RH. A total of three (GSH) conjugates (M2, M3 and M5) were detected in bile of rats treated with RH. We concluded that M2-M3 were directly derived from parent compound RH through spontaneous reaction with GSH. M5 was derived from M1 by reaction with GSH, which required cytoslic GSTs. M5 was further metabolized to the corresponding NAC conjugate (mercapturic acid) and was excreted in urine. P450 2C9 was mainly involved in the oxidation of RH.

Flagellar swimming in viscoelastic fluids: role of fluid elastic stress revealed by simulations based on experimental data.

Many important biological functions depend on microorganisms' ability to move in viscoelastic fluids such as mucus and wet soil. The effects of fluid elasticity on motility remain poorly understood, partly because the swimmer strokes depend on the properties of the fluid medium, which obfuscates the mechanisms responsible for observed behavioural changes. In this study, we use experimental data on the gaits of Chlamydomonas reinhardtii swimming in Newtonian and viscoelastic fluids as inputs to numerical simulations that decouple the swimmer gait and fluid type in order to isolate the effect of fluid elasticity on swimming. In viscoelastic fluids, cells employing the Newtonian gait swim faster but generate larger stresses and use more power, and as a result the viscoelastic gait is more efficient. Furthermore, we show that fundamental principles of swimming based on viscous fluid theory miss important flow dynamics: fluid elasticity provides an elastic memory effect that increases both the forward and backward speeds, and (unlike purely viscous fluids) larger fluid stress accumulates around flagella moving tangent to the swimming direction, compared with the normal direction.

Chemical Reactivity of Emodin and Its Oxidative Metabolites to Thiols.

Polygonum multiflorum is an herbal medicine widely employed in China. Hepatotoxicity of the herbal medicine has been well documented, but the mechanisms of the toxicity remain unknown. Emodin (EM) is a major constituent of the herb and has been reported to be hepatotoxic. The main purpose of this study was to define the metabolic pathways of EM in order to characterize the potential reactive intermediates. EM was incubated with rat liver microsomes or human liver microsomes, followed by LC-MS/MS analysis to investigate the in vitro and in vivo metabolism of EM. As a result, three monohydroxylation metabolites (M1-M3) were detected after exposure to EM: ω-hydroxyemodin, 2-hydroxyemodin, and 5-hydroxyemodin. Urinary M1 and M2 were detected in rats administered EM. Three mercapturic acids (M4-M6) were found in microsomal incubations containing EM, NADPH, and N-acetylcysteine. It appears that M4 originated from parent compound EM, and M5 and M6 originated from M1 and M2, respectively. Two biliary EM-derived GSH conjugates were found in EM-treated rats. One arose from direct adduction of EM with GSH, and the other was derived from M1. Cytochrome P450's 1A2, 2C19, and 3A4 were the predominant P450 enzymes to oxidize EM. The findings helped us to understand the mechanisms of EM-induced hepatotoxicity.

Hybrid structured fiber-optic Fabry-Perot interferometer for simultaneous measurement of strain and temperature.

We fabricate and experimentally demonstrate a hybrid structured Fabry-Perot interferometer (FPI) embedded in the middle of a fiber line for simultaneous measurement of axial strain and temperature. The FPI is composed of a silica-cavity cascaded to a spheroidal air-cavity, both of which are formed in a hollow annular core fiber (HACF). The fabrication process of the FPI includes only a fusion splice between a single-mode fiber and a HACF and several electrical arc discharges at the HACF near the splice point. Experimental results show that the strain and temperature sensitivities of the air-cavity can be 5.2 pm/µÎµ and 1.3 pm/C°, respectively, and those of the silica-cavity can be 1.1 pm/µÎµ and 13 pm/C°, respectively. The different sensitivities of silica-cavity and air-cavity to strain and temperature enable us to implement simultaneous sensing in strain and temperature.