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Background Recent reports of extreme levels of undersaturation in internal leaf air spaces have called into question one of the foundational assumptions of leaf gas exchange analysis, that leaf air spaces are effectively saturated with water vapour at leaf surface temperature. Historically, inferring the biophysical states controlling assimilation and transpiration from the fluxes directly measured by gas exchange systems has presented a number of challenges, including: (1) a mismatch in scales between the area of flux measurement, the biochemical cellular scale and the meso-scale introduced by the localization of the fluxes to stomatal pores; (2) the inaccessibility of the internal states of CO2 and water vapour required to define conductances; and (3) uncertainties about the pathways these internal fluxes travel. In response, plant physiologists have adopted a set of simplifying assumptions that define phenomenological concepts such as stomatal and mesophyll conductances. Scope Investigators have long been concerned that a failure of basic assumptions could be distorting our understanding of these phenomenological conductances, and the biophysical states inside leaves. Here we review these assumptions and historical efforts to test them. We then explore whether artefacts in analysis arising from the averaging of fluxes over macroscopic leaf areas could provide alternative explanations for some part, if not all, of reported extreme states of undersaturation. Conclusions Spatial heterogeneities can, in some cases, create the appearance of undersaturation in the internal air spaces of leaves. Further refinement of experimental approaches will be required to separate undersaturation from the effects of spatial variations in fluxes or conductances. Novel combinations of current and emerging technologies hold promise for meeting this challenge.

The entropy-driven monolayer assembly of hexagonal prisms and cylinders was studied under hard slit confinement. At the conditions investigated, the particles have two distinct and dynamically disconnected rotational states: unflipped and flipped, depending on whether their circular/hexagonal face is parallel or perpendicular to the wall plane. Importantly, these two rotational states cast distinct projection areas over the wall plane that favor either hexagonal or tetragonal packing. Monte Carlo simulations revealed a re-entrant melting transition where an intervening disordered Flipped-Unflipped (FUN) phase is sandwiched between a fourfold tetratic phase at high concentrations and a sixfold triangular solid at intermediate concentrations. The FUN phase contains a mixture of flipped and unflipped particles and is translationally and orientationally disordered. Complementary experiments were conducted with photolithographically fabricated cylindrical microparticles confined in a wedge cell. Both simulations and experiments show the formation of phases with comparable fraction of flipped particles and structure, i.e., the FUN phase, triangular solid, and tetratic phase, indicating that both approaches sample analogous basins of particle-orientation phase-space. The phase behavior of hexagonal prisms in a soft-repulsive wall model was also investigated to exemplify how tunable particle–wall interactions can provide an experimentally viable strategy to dynamically bridge the flipped and unflipped states.

Water acts as the solvent for natural biotic and abiotic processes and in many technological contexts. The availability of water to participate in chemical and physical processes is captured by thermodynamic variables which track the energetic state of water such as water activity and water potential. Our understanding of the energetic state of water in relevant processes is limited by a lack of sensors capable of providing accurate and reliable ex situ and in situ measurements of water activity. To address this technology gap, we present applications of a microtensiometer (μTM): a biomimetic microelectromechanical system (MEMS) sensor capable of measuring water activity in liquid, vapor, and semisolid (e.g., hydrogels, cheese) phases. We developed packaging, measurement systems, and methodology to enable us to make water activity measurements previously inaccessible to tensiometry. We present measurements in two contexts: (1) a small benchtop unit for ex situ measurements and (2) a probe format for in situ measurements. We demonstrate that the μTM can accurately measure water activity in a diversity of complex samples and agrees with chilled mirror hygrometry, an industry standard for water activity measurement.

International audience; Probing nanoconfined solutions in tortuous, mesoporous media is challenging because of pore size, complex pore connectivity, and the coexistence of multiple components and phases. Here, we use optical reflectance to experimentally investigate the wetting and drying of a mesoporous medium with ∼3-nm-diameter pores containing aqueous solutions of sodium chloride and lithium chloride. We show that the vapor activities (i.e., relative humidities) that correspond to optical features in the isotherms for solutions can be used to deduce the thermodynamic state of a nanoscopic solution that undergoes evaporation and crystallization upon drying and condensation and deliquescence when increasing the relative humidity. We emphasize specific equilibrium states of the system: the onset of draining during desorption and the end of filling during adsorption as well as percolation-induced scattering and crystallization. We find that theoretical arguments involving classical thermodynamics (a modified Kelvin–Laplace equation and classical nucleation theory) explain quantitatively the evolution of the optical features and thereby the state of the solution as a function of imposed vapor activity and solute concentration.

Significance Gaps in our ability to document local water relations in leaves compromise our ability to build complete models of leaf and plant function and our understanding of ecophysiological phenomena, such as response and adaptation to drought. Macroscopically, leaf water potential has been shown to impact vegetative growth and yield, susceptibility to disease, and, in extreme drought, plant viability, making it a promising candidate trait to improve water-use efficiency in plants. In this paper, we present a nanoscale sensor (AquaDust) that provides minimally disruptive measurements of water potential in leaves of intact plants at high spatial and temporal resolution. This creates opportunities for improving our understanding of the mechanisms coupling variations in water potential to biological and physical processes., Leaf water potential is a critical indicator of plant water status, integrating soil moisture status, plant physiology, and environmental conditions. There are few tools for measuring plant water status (water potential) in situ, presenting a critical barrier for developing appropriate phenotyping (measurement) methods for crop development and modeling efforts aimed at understanding water transport in plants. Here, we present the development of an in situ, minimally disruptive hydrogel nanoreporter (AquaDust) for measuring leaf water potential. The gel matrix responds to changes in water potential in its local environment by swelling; the distance between covalently linked dyes changes with the reconfiguration of the polymer, leading to changes in the emission spectrum via Förster Resonance Energy Transfer (FRET). Upon infiltration into leaves, the nanoparticles localize within the apoplastic space in the mesophyll; they do not enter the cytoplasm or the xylem. We characterize the physical basis for AquaDust’s response and demonstrate its function in intact maize (Zea mays L.) leaves as a reporter of leaf water potential. We use AquaDust to measure gradients of water potential along intact, actively transpiring leaves as a function of water status; the localized nature of the reporters allows us to define a hydraulic model that distinguishes resistances inside and outside the xylem. We also present field measurements with AquaDust through a full diurnal cycle to confirm the robustness of the technique and of our model. We conclude that AquaDust offers potential opportunities for high-throughput field measurements and spatially resolved studies of water relations within plant tissues.

Irrigation takes considerable amount of water; however, in many cases up to half of it is wasted. Improving the efficiency of irrigation control, therefore, is an important task for sustainable water management. Most existing irrigation control systems are based on soil moisture level. In this work, we explore theoretically the use of continuous values stem water potential (SWP) as a basis for control. SWP is a more direct measure of plant water status than the soil moisture level. After linearizing and discretizing a nonlinear dynamic model of water dynamics in a plant, we develop a model predictive control (MPC) framework for regulating SWP. To prevent plants from suffering water stress, data-driven robust MPC (DDRMPC) which captures the uncertainty of weather forecast error is implemented. A case study based on almond tree is presented to characterize the effectiveness of the DDRMPC strategy relative to on-off control. Sensitivity analysis on the prediction horizon and penalty weights were performed to investigate the varying irrigation control decisions. For the case characterized, the analysis shows that controlling tree trunk water potential through DDRMPC can save 2.5% amount of water comparing to on-off control while maintaining zero violation.

Leaf water potential is a critical indicator of plant water status, integrating soil moisture status, plant physiology, and environmental conditions. There are few tools for measuring plant water status (water potential) in situ, presenting a critical barrier for the development of appropriate phenotyping (measurement) methods for crop development and modeling efforts aimed at understanding water transport in plants. Here, we present the development of an in situ, minimally-disruptive hydrogel nanoreporter (AquaDust) for measuring leaf water potential. The gel matrix responds to changes in water potential in its local environment by swelling; the distance between covalently linked dyes changes with the reconfiguration of the polymer, leading to changes in the emission spectrum via Fluorescence Resonance Energy Transfer (FRET). Upon infiltration into leaves, the nanoparticles localize within the apoplastic space in the mesophyll; they do not enter the cytoplast or the xylem. We characterize the physical basis for AquaDust’s response and demonstrate its function in intact maize (Zea mays L.) leaves as a reporter of leaf water potential. We use AquaDust to measure gradients of water potential along intact, actively transpiring leaves as a function of water status; the localized nature of the reporters allows us to define a hydraulic model that distinguishes resistances inside and outside the xylem. We also present field measurements with AquaDust through a full diurnal cycle to confirm the robustness of the technique and of our model. We conclude that AquaDust offers potential opportunities for high-throughput, field measurements and spatially resolved studies of water relations within plant tissues.

Long after CO2 is injected into a brine aquifer, most reservoir-scale fluid dynamic simulations predict large fractions of the original plume will become immobilized via capillary trapping and dispersed throughout the formation. We begin our analysis with a reservoir in this state and consider the effects caused by a depressurization of the domain (e.g. from a nearby production well or newly formed fracture between neighboring reservoirs) and predict the fraction of CO2 that will be remobilized as a result. We then model the dynamics of this remobilized CO2 in two distinct steps: (1) vertical rise within the reservoir, followed by (2) spreading of mobile CO2 into the far-field of the domain and justify this approach from a scaling analysis of the governing equations. We show that a model of relative permeability that takes account of insights from percolation theory near the minimum CO2 saturation leads to much more rapid rise and subsequent radial spreading of remobilized CO2 than a traditional empirical correlation such as the Brooks-Corey model. Furthermore, we find that over a broad range of remobilized CO2 mass fraction and Bond number, the radial extent of the mobile plume does not exceed a factor of 1.8 times the radius of the original immobilized CO2 region.

A ring with a cross-section that has a blunt inner and sharper outer edge can attain an equilibrium orientation in a Newtonian fluid subject to a low Reynolds number simple shear flow. This may be contrasted with the continuous rotation exhibited by most rigid bodies. Such rings align along an orientation when the rotation due to fluid vorticity balances the counter-rotation due to the extensional component of the simple shear flow. While the viscous stress on the particle tries to rotate it, the pressure can generate a counter-vorticity torque that aligns the particle. Using boundary integral computations, we demonstrate ways to effectively control this pressure by altering the geometry of the ring cross-section, thus leading to alignment at moderate particle aspect ratios. Aligning rings that lack fore–aft symmetry can migrate indefinitely along the gradient direction. This differs from the periodic spatial trajectories of fore–aft asymmetric axisymmetric particles that rotate in periodic orbits. The mechanism for migration of aligned rings along the gradient direction is elucidated in this work. The migration speed can be controlled by varying the cross-sectional shape and size of the ring. Our results provide new insights into controlling motion of individual particles and thereby open new pathways towards manipulating macroscopic properties of a suspension.

In many species, sucrose en route out of the leaf migrates from photosynthetically active mesophyll cells into the phloem down its concentration gradient via plasmodesmata, i.e., symplastically. In some of these plants the process is entirely passive, but in others phloem sucrose is actively converted into larger sugars, raffinose and stachyose, and segregated (trapped), thus raising total phloem sugar concentration to a level higher than in the mesophyll. Questions remain regarding the mechanisms and selective advantages conferred by both of these symplastic loading processes. Here we present an integrated model - including local and global transport and the kinetics of oligomerization - for passive and active symplastic loading. We also propose a physical model of transport through the plasmodesmata. With these models, we predict that: 1) relative to passive loading, oligomerization of sucrose in the phloem, even in the absence of segregation, lowers the sugar content in the leaf required to achieve a given export rate and accelerates export for a given concentration of sucrose in the mesophyll; and 2) segregation of oligomers and the inverted gradient of total sugar content can be achieved for physiologically reasonable parameter values, but even higher export rates can be accessed in scenarios in which polymers are allowed to diffuse back into the mesophyll. We discuss these predictions in relation to further studies aimed at the clarification of loading mechanisms, fitness of active and passive symplastic loading, and potential targets for engineering improved rates of export.

The patterned differentiation of endothelial cells into tip and stalk cells represents an important step in the process of angiogenic sprouting. Vascular biologists hypothesize that changes in the density and overall structure of the vasculature can be traced in part to changes in the number of tip cells selected in the endothelium prior to sprout formation. However, the dominant hypotheses for tip cell selection invoke lateral inhibition via Notch; this juxtacrine mechanism predicts that a fixed fraction of endothelial cells become tip cells through a pattern-forming instability. Here, we present and analyze a hypothetical mechanism for tip cell selection that is based on endothelial competition for diffusible vascular endothelial growth factor (VEGF); this mechanism predicts that variable densities of tip cells emerge depending on the local (paracrine) production rate of VEGF. First, we hypothesize a network of VEGF signaling and trafficking based on previous experimental findings that could allow internalization of VEGF to occur with positive feedback. We formalize the hypothesis into a set of nonlinear ordinary differential equations and perform linear stability analysis to elucidate a general criterion for tip cell pattern formation under the mechanism. We use numerical integration to explore the nonlinear dynamics and final steady-states of tip cell patterns under this mechanism; the observed density of tip cells can be tuned from 10% to 84%. We conclude with proposals of future experiments and computational studies to explore how competitive consumption of diffusible VEGF may play a role in determining vascular structure.Statement of SignificanceThe patterned differentiation of endothelial cells into tip and stalk cells represents an important step in the process of blood vessel growth. Vascular biologists hypothesize that changes in the density and overall structure of the vasculature can be traced in part to changes in the number of tip cells selected during angiogenesis. However, the dominant hypotheses for tip cell selection predict that a locally fixed fraction of endothelial cells become tip cells following stimulation by vascular endothelial growth factor (VEGF). Here, we present and analyze a hypothetical mechanism for tip cell selection based on endothelial competition for diffusible VEGF; this mechanism predicts that variable densities of tip cells emerge depending on the local production rate of VEGF.

International audience; We investigate the filling and emptying of extreme ink-bottle porous media—micrometer-scale pores connected by nanometer-scale pores—when changing the pressure of the external vapor, in a case where the pore liquid contains solutes. These phenomena are relevant in diverse contexts, such as the weathering of building materials and artwork, aerosol formation in the atmosphere, and the hydration of soils and plants. Using model systems made of vein-shaped microcavities interconnected by a mesoporous matrix, we show experimentally that the presence of a nonvolatile solute shifts the condensation and evaporation transitions and in a way that is consistent with a modified Kelvin–Laplace equation that takes into account the osmotic pressure of the solution. Emptying occurs far below saturation, when the Kelvin stress, mediated by the large curvature of the liquid–vapor interfaces in the nanopores, is negative enough to induce spontaneous bubble nucleation in the microveins. Filling, on the other hand, occurs close to equilibrium (i.e., at saturation, psat for pure water and ps < psat for a solution), driven by the weak capillary pressure of the liquid–vapor interface in the microveins. Interestingly, solutes allow the system to reach situations where the vapor is supersaturated with respect to the solution (ps < p < psat). We show that in that latter situation, a condensation layer covers the outer surface of the porous system, preventing the generation of Kelvin stresses but inducing osmotic stresses and flows that are vapor pressure-dependent. The timescales and dynamics reflect these different driving forces: emptying proceeds through discrete, stochastic nucleation events with very fast, unsteady bubble growth associated with a poroelastic relaxation process, while filling occurs collectively in all veins of the sample through a slower steady-state process driven by a combination of osmosis and capillarity. The dynamics can however be rendered symmetrical between filling and emptying if bubbles pre-exist during emptying, a case that we explore using cycling of the vapor pressure around equilibrium.

In this paper, we investigate quantitatively the coupling between gradients of temperature and of chemical or water potential under steady state conditions in the vapor phase. This coupling is important for the measurement and modeling of the dynamics of water in unsaturated environments like soils and plants. We focus on a simple non-equilibrium scenario in which a gradient of temperature exists across an air-filled gap that separates two aqueous phases with no net transfer of water. This scenario is relevant for measurements of the water potential in environmental and industrial contexts. We use a new tool, a microtensiometer, to perform these measurements. We observed variations of water potential with difference of temperature across the air gap of $-\SI{7.9(3)}{\mega\pascal\per\kelvin}$, in agreement with previous measurements. Our result is close to a first order theoretical prediction, highlighting that most of the effect comes from the variation of saturation pressure with temperature. We then show that thermodiffusion (Soret effect) coupled to natural convection could occur in our experiment and discuss how these effects could explain the small discrepancy observed between measurements and first order theoretical prediction., * Version of January 2021: various clarifications * Early version contained mistakes: use of Philip and de Vries model was incorrect. New version gives updated values (using more accurate data from literature) and more detailed model, considering possible effects of thermodiffusion and natural convection in the experiment. Emphasis on the soil physics context is decreased

A time dependent injection strategy for greatly accelerating the immobilization of geologically sequestered CO 2 is proposed and analyzed. The injection of high density CO 2 into a brine aquifer is followed by brine flooding facilitating residual trapping and dissolution of the CO 2 on time scales much shorter than those that would occur by natural processes. One-dimensional kinematic wave equations are derived for the two-phase flow of brine and CO 2 and for the transport of dissolved CO 2 . A solution of these equations using the method of characteristics reveals that brine flooding is most effective when the kinematic wave speed of CO 2 saturation is higher than the propagation velocity of a shock wave separating the two-phase flow from the native brine. Finite volume simulation using the reservoir simulator TOUGH2 with PetraSIM interface are generally in good agreement with the one-dimensional model, but show that gravitational overriding of the CO 2 can become important if the duration of the injection process is too long. Both methods show that brine flooding is able to reduce the mass fraction of mobile CO 2 to less than 10% using a volume ratio brine:CO 2 of less than 2.75 on time scales comparable to that of the CO 2 injection.

We experimentally investigate the dynamics of capillary-driven flows at the nanoscale, using an original platform that combines nanoscale pores and microfluidic features. Our results show a coherent picture across multiple experiments including imbibition, poroelastic transient flows, and a drying-based method that we introduce. In particular, we exploit extreme drying stresses - up to 100 MPa of tension - to drive nanoflows and provide quantitative tests of continuum theories of fluid mechanics and thermodynamics (e.g. Kelvin-Laplace equation) across an unprecedented range. We isolate the breakdown of continuum as a negative slip length of molecular dimension., Comment: 5 pages; 4 figures

Adipose-derived stem cells (ASCs) are key regulators of new blood vessel formation and widely investigated for their role in tissue regeneration and tumorigenesis. However, the cellular and molecular mechanisms through which ASCs regulate angiogenesis are not well understood. Here, it was our goal to test the functional contribution of ASC-mediated extracellular matrix (ECM) remodeling on endothelial cell invasion. To isolate the effect of ECM-remodeling, ASCs were embedded within 3-D collagen type I hydrogels and pre-cultured for 7 days; controls were not pre-cultured. A confluent monolayer of human umbilical vein endothelial cells (HUVECs) was seeded on top and its invasion into the underlying hydrogel was analyzed. Without pre-culture, ASCs inhibited vascular sprouting by stabilizing the endothelium. In contrast, 7 day pre-culture of ASCs drastically increased invasion by HUVECs. This effect was largely mediated by proteolytic ECM degradation by ASC-derived matrix metalloproteinases (MMPs) rather than vascular endothelial growth factor (VEGF), as our results indicated that blockade of MMPs, but not VEGF, inhibited endothelial sprouting. Collectively, these data suggest that the angiogenic capability of ASCs is modulated by their proteolytic remodeling of the ECM, opening new avenues for pro- and anti-angiogenic therapies.

Cancer metabolism has received renewed interest as a potential target for cancer therapy. In this study, we use a multi-scale modeling approach to interrogate the implications of three metabolic scenarios of potential clinical relevance: the Warburg effect, the reverse Warburg effect and glutamine addiction. At the intracellular level, we construct a network of central metabolism and perform flux balance analysis (FBA) to estimate metabolic fluxes; at the cellular level, we exploit this metabolic network to calculate parameters for a coarse-grained description of cellular growth kinetics; and at the multicellular level, we incorporate these kinetic schemes into the cellular automata of an agent-based model (ABM), iDynoMiCS. This ABM evaluates the reaction-diffusion of the metabolites, cellular division and motion over a simulation domain. Our multi-scale simulations suggest that the Warburg effect provides a growth advantage to the tumor cells under resource limitation. However, we identify a non-monotonic dependence of growth rate on the strength of glycolytic pathway. On the other hand, the reverse Warburg scenario provides an initial growth advantage in tumors that originate deeper in the tissue. The metabolic profile of stromal cells considered in this scenario allows more oxygen to reach the tumor cells in the deeper tissue and thus promotes tumor growth at earlier stages. Lastly, we suggest that glutamine addiction does not confer a selective advantage to tumor growth with glutamine acting as a carbon source in the tricarboxylic acid (TCA) cycle, any advantage of glutamine uptake must come through other pathways not included in our model (e.g., as a nitrogen donor). Our analysis illustrates the importance of accounting explicitly for spatial and temporal evolution of tumor microenvironment in the interpretation of metabolic scenarios and hence provides a basis for further studies, including evaluation of specific therapeutic strategies that target metabolism., Author summary Cancer metabolism is an emerging hallmark of cancer. In the past decade, a renewed focus on cancer metabolism has led to several distinct hypotheses describing the role of metabolism in cancer. To complement experimental efforts in this field, a scale-bridging computational framework is needed to allow rapid evaluation of emerging hypotheses in cancer metabolism. In this study, we present a multi-scale modeling platform and demonstrate the distinct outcomes in population-scale growth dynamics under different metabolic scenarios: the Warburg effect, the reverse Warburg effect and glutamine addiction. Within this modeling framework, we confirmed population-scale growth advantage enabled by the Warburg effect, provided insights into the symbiosis between stromal cells and tumor cells in the reverse Warburg effect and argued that the anaplerotic role of glutamine is not exploited by tumor cells to gain growth advantage under resource limitations. We point to the opportunity for this framework to help understand tissue-scale response to therapeutic strategies that target cancer metabolism while accounting for the tumor complexity at multiple scales.

Despite its relevance in numerous natural and industrial processes, the solubility of molecular oxygen has never been directly measured in capillary condensed liquid water. In this article, we measure oxygen solubility in liquid water trapped within nanoporous samples, in metastable equilibrium with a subsaturated vapor. We show that solubility increases two-fold at moderate subsaturations (RH ~ 0.55). This evolution with relative humidity is in good agreement with a simple thermodynamic prediction using properties of bulk water, previously verified experimentally at positive pressure. Our measurement thus verifies the validity of this macroscopic thermodynamic theory to strong confinement and large negative pressures, where ignificant non-idealities are expected. This effect has strong implications for important oxygen-dependent chemistries in natural and technological contexts., Accepted for publication in Langmuir

We develop a novel data-driven robust model predictive control (DDRMPC) approach for automatic control of irrigation systems. The fundamental idea is to integrate both mechanistic models, which describe dynamics in soil moisture variations, and data-driven models, which characterize uncertainty in forecast errors of evapotranspiration and precipitation, into a holistic systems control framework. To better capture the support of uncertainty distribution, we take a new learning-based approach by constructing uncertainty sets from historical data. For evapotranspiration forecast error, the support vector clustering-based uncertainty set is adopted, which can be conveniently built from historical data. As for precipitation forecast errors, we analyze the dependence of their distribution on forecast values, and further design a tailored uncertainty set based on the properties of this type of uncertainty. In this way, the overall uncertainty distribution can be elaborately described, which finally contributes to rational and efficient control decisions. To assure the quality of data-driven uncertainty sets, a training-calibration scheme is used to provide theoretical performance guarantees. A generalized affine decision rule is adopted to obtain tractable approximations of optimal control problems, thereby ensuring the practicability of DDRMPC. Case studies using real data show that, DDRMPC can reliably maintain soil moisture above the safety level and avoid crop devastation. The proposed DDRMPC approach leads to a 40% reduction of total water consumption compared to the fine-tuned open-loop control strategy. In comparison with the carefully tuned rule-based control and certainty equivalent model predictive control, the proposed DDRMPC approach can significantly reduce the total water consumption and improve the control performance.

Oxygen status and tissue dimensionality are critical determinants of tumor angiogenesis, a hallmark of cancer and an enduring target for therapeutic intervention. However, it is unclear how these microenvironmental conditions interact to promote neovascularization, due in part to a lack of comprehensive, unbiased data sets describing tumor cell gene expression as a function of oxygen levels within three-dimensional (3D) culture. Here, we utilized alginate-based, oxygen-controlled 3D tumor models to study the interdependence of culture context and the hypoxia response. Microarray gene expression analysis of tumor cells cultured in 2D versus 3D under ambient or hypoxic conditions revealed striking interdependence between culture dimensionality and hypoxia response, which was mediated in part by pro-inflammatory signaling pathways. In particular, interleukin-8 (IL-8) emerged as a major player in the microenvironmental regulation of the hypoxia program. Notably, this interaction between dimensionality and oxygen status via IL-8 increased angiogenic sprouting in a 3D endothelial invasion assay. Taken together, our data suggest that pro-inflammatory pathways are critical regulators of tumor hypoxia response within 3D environments that ultimately impact tumor angiogenesis, potentially providing important therapeutic targets. Furthermore, these results highlight the importance of pathologically relevant tissue culture models to study the complex physical and chemical processes by which the cancer microenvironment mediates new vessel formation.

Vascular plants rely on differences in osmotic pressure to export sugars from regions of synthesis (mature leaves) to sugar sinks (roots, fruits). In this process, known as Munch pressure flow, the loading of sugars from photosynthetic cells to the export conduit (the phloem) is crucial, as it sets the pressure head necessary to power long-distance transport. Whereas most herbaceous plants use active mechanisms to increase phloem sugar concentration above that of the photosynthetic cells, in most tree species, for which transport distances are largest, loading seems, counterintuitively, to occur by means of passive symplastic diffusion from the mesophyll to the phloem. Here, we use a synthetic microfluidic model of a passive loader to explore the non-linear dynamics that arise during export and determine the ability of passive loading to drive long-distance transport. We first demonstrate that in our device, the phloem concentration is set by the balance between the resistances to diffusive loading from the source and convective export through the phloem. Convection-limited export corresponds to classical models of Munch transport, where the phloem concentration is close to that of the source; in contrast, diffusion-limited export leads to small phloem concentrations and weak scaling of flow rates with hydraulic resistance. We then show that the effective regime of convection-limited export is predominant in plants with large transport resistances and low xylem pressures. Moreover, hydrostatic pressures developed in our synthetic passive loader can reach botanically relevant values as high as 10 bars. We conclude that passive loading is sufficient to drive long-distance transport in large plants, and that trees are well suited to take full advantage of passive phloem loading strategies.

In leaves, the transpirational flux of water exits the veins as liquid and travels toward the stomata in both the vapor and liquid phases before exiting the leaf as vapor. Yet, whether most of the evaporation occurs from the vascular bundles (perivascular), from the photosynthetic mesophyll cells, or within the vicinity of the stomatal pore (peristomatal) remains in dispute. Here, a one-dimensional model of the competition between liquid and vapor transport is developed from the perspective of nonisothermal coupled heat and water molecule transport in a composite medium of airspace and cells. An analytical solution to the model is found in terms of the energy and transpirational fluxes from the leaf surfaces and the absorbed solar energy load, leading to mathematical expressions for the proportions of evaporation accounted for by the vascular, mesophyll, and epidermal regions. The distribution of evaporation in a given leaf is predicted to be variable, changing with the local environment, and to range from dominantly perivascular to dominantly peristomatal depending on internal leaf architecture, with mesophyll evaporation a subordinate component. Using mature red oak (Quercus rubra) trees, we show that the model can be solved for a specific instance of a transpiring leaf by combining gas-exchange data, anatomical measurements, and hydraulic experiments. We also investigate the effect of radiation load on the control of transpiration, the potential for condensation on the inside of an epidermis, and the impact of vapor transport on the hydraulic efficiency of leaf tissue outside the xylem.

Plants live dangerously, but gracefully. To remain hydrated, they exploit liquid water in the thermodynamically metastable state of negative pressure, similar to a rope under tension. This tension allows them to pull water out of the soil and up to their leaves. When this liquid rope breaks, owing to cavitation, they catch the ends to keep it from unraveling and then bind it back together. In parallel, they operate a second vascular system for the circulation of metabolites though their tissues, this time with positive pressures and flow that passes from leaf to root. In this article, we review the current state of understanding of water management in plants with an emphasis on the rich coupling of transport phenomena, thermodynamics, and active biological processes. We discuss efforts to replicate plant function in synthetic systems and point to opportunities for physical scientists and engineers to benefit from and contribute to the study of plants.

Tensiometers sense the chemical potential of water (or water potential, Ψw) in an external phase of interest by measuring the pressure in an internal volume of liquid water in equilibrium with that phase. For sub-saturated phases, the internal pressure is below atmospheric and frequently negative; the liquid is under tension. Here, we present the initial characterization of a new tensiometer based on a microelectromechanical pressure sensor and a nanoporous membrane. We explain the mechanism of operation, fabrication, and calibration of this device. We show that these microtensiometers operate stably out to water potentials below -10 MPa, a tenfold extension of the range of current tensiometers. Finally, we present use of the device to perform an accurate measurement of the equation of state of liquid water at pressures down to -14 MPa. We conclude with a discussion of outstanding design considerations, and of the opportunities opened by the extended range of stability and the small form factor in sensing applications, and in fundamental studies of the thermodynamic properties of water.

The design and analysis of plant-inspired loop heat pipes (LHPs) that would exploit nanoporous membranes to allow for operation with large capillary pressures and superheated liquid are presented. The operating concepts of this superheated loop heat pipe (SHLHP) resemble the transpiration process in vascular plants: reduction of pressure in leaves drives sap flow up from the roots and overcomes gravity, viscous drag, and reduced chemical potential of water in subsaturated soils. We present a model for steady-state operation and a linear response analysis of both the conventional and superheated designs. Our analysis shows that these SHLHPs could: (1) extend the limitations of conventional LHPs imposed by thermodynamic properties of the working fluid, (2) provide efficient heat transfer over long distances and against large accelerations, and (3) allow for operation in a subsaturated state that would eliminate the thermal resistance and entrainment effect of the liquid film of conventional designs. © 2013 American Institute of Chemical Engineers AIChE J 60: 762–777, 2014

In perfusion experiments, the hydraulic conductance of stem segments () responds to changes in the properties of the perfusate, such as the ionic strength (), pH, and cationic identity. We review the experimental and theoretical work on this phenomenon. We then proceed to explore the hypothesis that electrokinetic effects in the bordered pit membrane (BPM) contribute to this response. In particular, we develop a model based on electroviscosity in which hydraulic conductance of an electrically charged porous membrane varies with the properties of the electrolyte. We use standard electrokinetic theory, coupled with measurements of electrokinetic properties of plant materials from the literature, to determine how the conductance of BPMs, and therefore , may change due to electroviscosity. We predict a nonmonotonic variation of with with a maximum reduction of 18%. We explore how this reduction depends on the characteristics of the sap and features of the BPM, such as pore size, density of chargeable sites, and their dissociation constant. Our predictions are consistent with changes in observed for physiological values of sap and pH. We conclude that electroviscosity is likely responsible, at least partially, for the electrolyte dependence of conductance through pits and that electroviscosity may be strong enough to play an important role in other transport processes in xylem. We conclude by proposing experiments to differentiate the impact of electroviscosity on from that of other proposed mechanisms.

Both physiological and pathological tissue remodeling (e.g., during wound healing and cancer, respectively) require new blood vessel formation via angiogenesis, but the underlying microenvironmental mechanisms remain poorly defined due in part to the lack of biologically relevant in vitro models. Here, we present a biomaterials-based microfluidic 3D platform for analysis of endothelial sprouting in response to morphogen gradients. This system consists of three lithographically defined channels embedded in type I collagen hydrogels. A central channel is coated with endothelial cells, and two parallel side channels serve as a source and a sink for the steady-state generation of biochemical gradients. Gradients of vascular endothelial growth factor (VEGF) promoted sprouting, whereby endothelial cell responsiveness was markedly dependent on cell density and vessel geometry regardless of treatment conditions. These results point toward mechanical and/or autocrine mechanisms that may overwhelm pro-angiogenic paracrine signaling under certain conditions. To date, neither geometrical effects nor cell density have been considered critical determinants of angiogenesis in health and disease. This biomimetic vessel platform demonstrated utility for delineating hitherto underappreciated contributors of angiogenesis, and future studies may enable important new mechanistic insights that will inform anti-angiogenic cancer therapy.

Most rigid, torque-free, low-Reynolds-number, axisymmetric particles undergo a time-periodic tumbling motion in a simple shear flow, with their axes of symmetry following a set of closed Jeffery orbits. We have identified a class of rigid, ring-like particles whose axes of symmetry instead reach a permanent alignment near the velocity gradient direction with the plane of the particle aligning near the flow–vorticity plane. An asymptotic analysis for small particle aspect ratio (ratio of length parallel to the axis of symmetry to diameter perpendicular to the axis) shows that an appropriate asymmetry of the ring cross-section with a thinner outer edge and thicker inner edge leads to a tendency to rotate in a direction opposite to the vorticity; this tendency can balance the usual rotation rate associated with the finite thickness of the particle. Boundary integral computations for finite particle aspect ratios are used to determine the conditions of aspect ratio and degree of asymmetry that lead to the aligning behaviour and the final orientation of the axis of symmetry of the aligned particles. The aligning particle follows an equation of motion similar to the Leslie–Erickson equation for the director of a small-molecule nematic liquid crystal. However, whereas the alignment of the director arises from intermolecular interactions, the ring-like particle aligns solely due to its intrinsic rotational motion in a low-Reynolds-number flow.

International audience; Liquid can sustain mechanical tension as its pressure drops below the vapor-liquid coexistence line and becomes less than zero, until it reaches the stability limit – the pressure at which cavitation (i.e. the nucleation of vapor bubbles in bulk liquid) inevitably occurs. For liquid water, its stability limit is still a subject of debate: the results obtained by researchers using a variety of techniques show discrepancies between the values of stability limit and its temperature-dependence as temperature approaches 0°C. In this work, we present a study of the stability limit of water with the metastable vapor-liquid equilibrium (MVLE) method in which a volume of liquid is equilibrated with its unsaturated vapor via nanoporous silicon membrane. We also report on an experimental system which enables test of the temperature-dependence of the stability limit with MVLE. Our results falls in the range between -20 and -30 MPa; a range that is in consistent with the majority of the experiments but is far less negative than the limit obtained in experiments involving quartz inclusions and that predicted for homogeneous nucleation. Further, the stability limit we found increases monotonically (larger tension) as temperature approaches 0°C; this trend contradicts the centrifugal result of Briggs but agrees with the experiments by acoustic cavitation.

We study the spatio-temporal dynamics of water uptake by capillary condensation from unsaturated vapor in mesoporous silicon layers (pore radius $r_\mathrm{p} \simeq 2$ nm), taking advantage of the local changes in optical reflectance as a function of water saturation. Our experiments elucidate two qualitatively different regimes as a function of the imposed external vapor pressure: for low saturations, equilibration occurs via a diffusion-like process; for high saturations, an imbibition-like wetting front results in fast equilibration towards a fully saturated sample. We show that the imbibition dynamics can be described by a modified Lucas-Washburn equation that takes into account the liquid stresses implied by Kelvin equation., Comment: 11 pages, 5 figures, under review

Breast cancer initiation, invasion and metastasis span multiple length and time scales. Molecular events at short length scales lead to an initial tumorigenic population, which left unchecked by immune action, acts at increasingly longer length scales until eventually the cancer cells escape from the primary tumor site. This series of events is highly complex, involving multiple cell types interacting with (and shaping) the microenvironment. Multiscale mathematical models have emerged as a powerful tool to quantitatively integrate the convective-diffusion-reaction processes occurring on the systemic scale, with the molecular signaling processes occurring on the cellular and subcellular scales. In this study, we reviewed the current state of the art in cancer modeling across multiple length scales, with an emphasis on the integration of intracellular signal transduction models with pro-tumorigenic chemical and mechanical microenvironmental cues. First, we reviewed the underlying biomolecular origin of breast cancer, with a special emphasis on angiogenesis. Then, we summarized the development of tissue engineering platforms which could provide high-fidelity ex vivo experimental models to identify and validate multiscale simulations. Lastly, we reviewed top-down and bottom-up multiscale strategies that integrate subcellular networks with the microenvironment. We present models of a variety of cancers, in addition to breast cancer specific models. Taken together, we expect as the sophistication of the simulations increase, that multiscale modeling and bottom-up agent-based models in particular will become an increasingly important platform technology for basic scientific discovery, as well as the identification and validation of potentially novel therapeutic targets.

In many important chemical processes, the laminar flow regime is inescapable and defines the performance of reactors, separators, and analytical instruments. In the emerging field of microchemical process or lab-on-a-chip, this constraint is particularly rigid. Here, we review developments in the use of chaotic laminar flows to improve common transport processes in this regime. We focus on four: mixing, interfacial transfer, axial dispersion, and spatial sampling. Our coverage demonstrates the potential for chaos to improve these processes if implemented appropriately. Throughout, we emphasize the usefulness of familiar theoretical models of transport for processes occurring in chaotic flows. Finally, we point out open challenges and opportunities in the field.

Microvascular networks support metabolic activity and define microenvironmental conditions within tissues in health and pathology. Recapitulation of functional microvascular structures in vitro could provide a platform for the study of complex vascular phenomena, including angiogenesis and thrombosis. We have engineered living microvascular networks in three-dimensional tissue scaffolds and demonstrated their biofunctionality in vitro. We describe the lithographic technique used to form endothelialized microfluidic vessels within a native collagen matrix; we characterize the morphology, mass transfer processes, and long-term stability of the endothelium; we elucidate the angiogenic activities of the endothelia and differential interactions with perivascular cells seeded in the collagen bulk; and we demonstrate the nonthrombotic nature of the vascular endothelium and its transition to a prothrombotic state during an inflammatory response. The success of these microvascular networks in recapitulating these phenomena points to the broad potential of this platform for the study of cardiovascular biology and pathophysiology.

We present the development and characterization of nanoparticles loaded with a custom phosphor; we exploit these nanoparticles to perform quantitative measurements of the concentration of oxygen within three-dimensional (3-D) tissue cultures in vitro and blood vessels in vivo. We synthesized a customized ruthenium (Ru)-phosphor and incorporated it into polymeric nanoparticles via self-assembly. We demonstrate that the encapsulated phosphor is non-toxic with and without illumination. We evaluated two distinct modes of employing the phosphorescent nanoparticles for the measurement of concentrations of oxygen: 1) in vitro, in a 3-D microfluidic tumor model via ratiometric measurements of intensity with an oxygen-insensitive fluorophore as a reference, and 2) in vivo, in mouse vasculature using measurements of phosphorescence lifetime. With both methods, we demonstrated micrometer-scale resolution and absolute calibration to the dissolved oxygen concentration. Based on the ease and customizability of the synthesis of the nanoparticles and the flexibility of their application, these oxygen-sensing polymeric nanoparticles will find a natural home in a range of biological applications, benefiting studies of physiological as well as pathological processes in which oxygen availability and concentration play a critical role.

Rapid, effective host cell invasion and vascularization is essential for durable incorporation of avascular tissue-replacement scaffolds. In this study, the authors sought to qualitatively and quantitatively determine which of two commercially available products (i.e., Strattice and Integra) facilitates more rapid cellular and vascular invasion in a murine model of graft incorporation.Integra and Strattice were implanted subcutaneously into the dorsa of C57BL/6 mice; harvested after 3, 7, or 14 days; and stained with hematoxylin and eosin, 4',6-diamidino-2-phenylindole, and immunohistochemical stains for CD31 and α-smooth muscle actin. Exponential decay equations describing cellular invasion through each layer were fit to each material/time point. Mean cell density and cell frequency maps were created denoting extent of invasion by location within the scaffold.Qualitative analysis demonstrated extensive cellular infiltration into Integra by 3 days and increasing over the remaining 14 days. Invasion of Strattice was sparse, even after 14 days. α-Smooth muscle actin immunohistochemistry revealed blood vessel formation within Integra by 14 days but no analogous neovascularization in Strattice. Mean decay equations for Integra and Strattice were y = 76.3(0.59) and y = 75.5(0.33), respectively. Both cell density measurements and frequency mapping demonstrated that, at all time points, Integra manifested a greater density/depth of cellular invasion when compared with Strattice.These data confirm empiric clinical observations that Integra is more rapidly invaded than Strattice when placed in a suitable host bed. A remnant microvasculature template is not sufficient for effective cellular ingrowth into an artificial tissue construct. These findings provide insight into means for improving future dermal replacement products.

The collision of particles influences the behavior of suspensions through the formation of aggregates for adhesive particles or through the contributions of solid-body contacts to the stress for nonadhesive particles. The simplest estimate of the collision rate, termed the ideal collision rate, is obtained when particles translate and rotate with the flow but have no hydrodynamic or colloidal interactions. Smoluchowski calculated the ideal collision frequency of spherical particles in 1917. So far, little work has been done to understand rate of collision for nonspherical particles. In this work, we calculate the ideal collision rate for cylindrical particles over a broad range of particle aspect ratios r defined as the ratio of length to diameter. Monte Carlo simulations are performed with initial relative positions and orientations that model the rate of approach of noninteracting particles following Jeffery orbits with several choices of the orbit distribution. The role of rotational motion of particles on collision frequency is elucidated by comparing the ideal collision rate calculations with similar calculations for nonrotating particles. It is shown that the ratio of the collision rate of cylinders to that of spheres that circumscribe the cylinders is proportional to 1/rr(e) for r ≫ 1 and r(e) for r ≪ 1. Here, r(e) is the effective aspect ratio defined as the aspect ratio of a spheroid having the same period of rotation as the cylinder. The effective aspect ratio of the cylindrical particles was determined using finite element calculations of the torque on nonrotating cylinders with their axes parallel to the velocity and velocity gradient directions. In addition to deriving the total collision rate, we categorize collisions as side-side, edge-side, and face-edge based on the initial point of contact. Most collisions are found to be side-edge for r ≫ 1 and face-edge for r ≪ 1, suggesting that nonlinear aggregates will develop if particles stick at the point of first contact.

Tissue templates for reconstruction and regeneration in vivo have achieved clinical successes for homogeneous tissues in well vascularized regions. Outstanding challenges exist for applications in poorly vascularized regions and for spatially differentiated tissues. Here, we present a strategy to control the spatial patterns of cell and vascular ingrowth throughout the volume of a bioremodelable and resorbable matrix via well-defined micropores and networks of microchannels. Our presentation of this approach includes: a description of a lithographic technique to form deterministic microstructures within a matrix of native collagen; elucidation of multistep process by which microstructures drive rapid invasion and vascularization; and demonstration of long range guidance of invasion through the full thickness of patterned templates. Experiments were performed in a murine wound model. These microstructured tissue templates (MTTs) could improve outcomes in reconstructive surgery and open paths to directed regeneration of spatially heterogeneous tissues or organs.

Oxygen (O2) reduction has long been the factor limiting the power density of most fuel cells. Membraneless, microfluidic fuel cells are a promising new fuel cell technology, yet they are affected e...

Type I collagen is a favorable substrate for cell adhesion and growth and is remodelable by many tissue cells; these characteristics make it an attractive material for the study of dynamic cellular processes. Low mass fraction (1.0–3.0 mg/ml), hydrated collagen matrices used for three-dimensional cell culture permit cellular movement and remodeling, but their microstructure and mechanics fail to mimic characteristics of many extracellular matrices in vivo and limit the definition of fine-scale geometrical features (< 1 mm) within scaffolds. In this study, we worked with hydrated type I collagen at mass fractions between 3.0 and 20 mg/ml to define the range of densities over which the matrices support both microfabrication and cellular remodeling. We present pore and fiber dimensions based on confocal microscopy and longitudinal modulus and hydraulic permeability based on confined compression. We demonstrate faithful reproduction of simple pores of 50 µm-diameter over the entire range and formation of functional microfluidic networks for mass fractions greater than 10.0 mg/ml. We present quantitative characterization of the rate and extent of cellular remodelability using human umbilical vein endothelial cells. Finally, we present a co-culture with tumor cells and discuss the implications of integrating microfluidic control within scaffolds as a tool to study spatial and temporal signaling during tumor angiogenesis and vascularization of tissue-engineered constructs.

Plant scientists believe that transpiration-the motion of water from the soil, through a vascular plant, and into the air-occurs by a passive, wicking mechanism. This mechanism is described by the cohesion-tension theory: loss of water by evaporation reduces the pressure of the liquid water within the leaf relative to atmospheric pressure; this reduced pressure pulls liquid water out of the soil and up the xylem to maintain hydration. Strikingly, the absolute pressure of the water within the xylem is often negative, such that the liquid is under tension and is thermodynamically metastable with respect to the vapour phase. Qualitatively, this mechanism is the same as that which drives fluid through the synthetic wicks that are key elements in technologies for heat transfer, fuel cells and portable chemical systems. Quantitatively, the differences in pressure generated in plants to drive flow can be more than a hundredfold larger than those generated in synthetic wicks. Here we present the design and operation of a microfluidic system formed in a synthetic hydrogel. This synthetic 'tree' captures the main attributes of transpiration in plants: transduction of subsaturation in the vapour phase of water into negative pressures in the liquid phase, stabilization and flow of liquid water at large negative pressures (-1.0 MPa or lower), continuous heat transfer with the evaporation of liquid water at negative pressure, and continuous extraction of liquid water from subsaturated sources. This development opens the opportunity for technological uses of water under tension and for new experimental studies of the liquid state of water.

Most methods to culture cells in three dimensions depend on a cell-seedable biomaterial to define the global structure of the culture and the microenvironment of the cells. Efforts to tailor these scaffolds have focused on the chemical and mechanical properties of the biomaterial itself. Here, we present a strategy to control the distributions of soluble chemicals within the scaffold with convective mass transfer via microfluidic networks embedded directly within the cell-seeded biomaterial. Our presentation of this strategy includes: a lithographic technique to build functional microfluidic structures within a calcium alginate hydrogel seeded with cells; characterization of this process with respect to microstructural fidelity and cell viability; characterization of convective and diffusive mass transfer of small and large solutes within this microfluidic scaffold; and demonstration of temporal and spatial control of the distribution of non-reactive solutes and reactive solutes (that is, metabolites) within the bulk of the scaffold. This approach to control the chemical environment on a micrometre scale within a macroscopic scaffold could aid in engineering complex tissues.

Motivated by the necessity to engineer appropriately stratified cartilage, the shear mechanics of layered, bovine chondrocyte-seeded 20 mg/mL alginate scaffolds were investigated and related to the structure and biochemical composition. Chondrocyte-seeded alginate scaffolds were exposed to a calcium-chelating solution, layered, crosslinked in CaCl 2 , and cultured for 10 weeks. The shear mechanical properties of the layered gels were statistically similar to those of the non-layered controls. Shear modulus of layered gels increased by approximately six-fold while toughness and shear strength increased by more than two-fold during the culture period. Hydroxyproline content in both layered gels and controls had statistically significant increases after 6 weeks. Glycosaminoglycan (GAG) content of controls increased throughout culture while GAG content in layered gels leveled off after 4 weeks. Hematoxylin and eosin histological staining showed tissue growth at the interface over the first 4 weeks. Shear mechanical properties in the engineered tissues showed significant correlations to hydroxyproline content. Dependence of interfacial mechanical properties on hydroxyproline content was most evident for layered gels when compared to controls, especially for toughness and shear strength. Additionally, interfacial properties showed almost no dependence on GAG content. These findings demonstrate the feasibility of creating stratified engineered tissues through layering and that collagen deposition is necessary for interfacial integrity.

We review recent research that involves the interaction of nanomaterials such as nanoparticles, nanowires, and carbon nanotubes with proteins. We begin with a focus on the fundamentals of the structure and function of proteins on nanomaterials. We then review work in three areas that exploit these interactions: (1) sensing, (2) assembly of nanomaterials by proteins and proteins by nanomaterials, and (3) interactions with cells. We conclude with the identification of challenges and opportunities for the future.

Vascular anastomosis – the fusion of vessels from two distinct branches of the vascular system – represents a critical step in vascular growth under both healthy and pathological conditions, in vivo, and presents an important target for engineering of vascularized tissues, in vitro. Recent works in animal models have advanced our understanding of the molecular and cellular players in vascular anastomosis, but questions remain related to cellular dynamics and control of this process, in vitro. In this study, we exploited a three-dimensional (3-D) culture platform to examine the dynamics of endothelial cell (EC) during and after vascular anastomosis by allowing angiogenesis and vasculogenesis to proceed in parallel. We show that anastomosis occurs between sprouts formed by angiogenesis from an endothelium and tubes formed by vasculogenesis in the bulk of a 3-D matrix. This fusion leads to highly connected vessels that span from the surface of the matrix into the bulk in a manner that depends on cell density and identity. Further, we observe and analyze intermixing of endothelial cells of distinct origin (surface versus bulk) within the vessels structures that are formed; we provide evidence that the cells migrate along pre-existing vessels segments as part of this intermixing process. We conclude that anastomosis can occur between vessels emerging by angiogenesis and vasculogenesis and that this process may play an important role in contexts such as wound healing.