Air spread through a wetted deformable membrane: Implications for the mechanism of soft valves in plants.

Plants have a special structure, torus-margo (TM) pit, which comprises a thickened torus at the center encircled by a highly porous margo. It is regarded as a key evolutionary structure to enable stable water transport, minimizing the air spread in the vessels. However, its valve-like dynamics to regulate two-phase flows still remains unclear even at a single pit level. Here, we study the air spreading dynamics using a bioinspired model of this soft pit valve. We divide it into the initial onset and the consecutive air-spreads, and propose the criteria of TM structures as the valve-like function. To delay the onset of air spread, the margo region should be thin and deformable enough to seal the pit aperture with the torus before the air penetration. Even after the onset, the membranes whose maximum pore size is smaller than its thickness can avoid continuous air-spread. The criteria also fit properly into botanical data on the morphologies of TM pits, implying that their valve-like behaviors may alleviate the tradeoff between hydraulic safety and efficiency at the single pit level. Our study would help to understand of the mechanistic pit-level strategy and also can provide insight into fluidic systems to control interfacial phenomena.

Air spreading through wetted cellulose membranes: Implications for the safety function of hydraulic valves in plants.

Plants transport water against the risk of cavitation inside xylem vessels, called "embolism." As one of their hydraulic strategies, pit membranes composed of cellulose fibers have been known as safety valves that prevent the spreading of embolism towards adjacent xylem vessels. However, detailed observation of embolism spreading through a pit membrane is still lacking. Here, we hypothesized that the pit membranes normally remain to be wetted in xylem vessels and noticed in particular the hydraulic role of water film on air spreading that has been overlooked previously. For the hydrodynamic study of the embolism spreading through a wetted pit membrane, we investigated the penetration and spreading dynamics of air plugs through the wetted cellulose membrane in a channel flow. Air spreading exhibits two types of dynamics: continuous and discrete spreading. We elucidated the correlation of dynamic characteristics of air flow and pressure variations according to membrane thickness. Our study speculates that the thickness of pit membranes affects the behaviors of water film captured by cellulose fibers, and it is a crucial criterion for the reversible gating of further spreading of embolism throughout xylem networks.

Penetration of a bubble through porous membranes with different wettabilities.

Porous structures with various surface wettabilities have been used to handle gas bubbles underwater for practical applications, such as separation, collection, detachment, and migration of the bubbles. Despite the increasing interest in porous structures, the effects of surface wettability on the behaviors of bubbles at porous surfaces have not been fully understood. Herein, we aim to examine the entire dynamics from collision to disappearance of a bubble through a porous membrane with different surface wettabilities. We divided the dynamics into three stages based on the characteristic behaviors such as bubble bouncing and contact line variation. Bubble dynamics is dominated by the existence of air layers covering the membrane surface. Bubbles on hydrophilic and hydrophobic membranes, which do not retain air layer, show the same removal pattern; they bounce on the surfaces, and then penetrate the membranes with pinned and moving contact line in sequence. In contrast, bubbles immediately penetrate the superhydrophobic membrane following the spread along the air layer. The characteristic time for bubble removal depends on the wettability, which affects the membrane permeability. The experimental characterization and theoretical analysis achieved in this work would improve the physical understanding of bubble dynamics on porous membranes and allow a proper design in bubble-related applications.

Removal of fine particulate matter (PM2.5) via atmospheric humidity caused by evapotranspiration.

Reduction of particulate matter (PM) has emerged as one of the most significant challenges in public health and environment protection worldwide. To address PM-related problems and effectively remove fine particulate matter (PM2.5), environmentalists proposed tree planting and afforestation as eco-friendly strategies. However, the PM removal effect of plants and its primary mechanism remains uncertain. In this study, we experimentally investigated the PM removal performance of five plant species in a closed chamber and the effects of relative humidity (RH) caused by plant evapotranspiration, as a governing parameter. On the basis of the PM removal test for various plant species, we selected Epipremnum aureum (Scindapsus) as a representative plant to identify the PM removal efficiency depending on evapotranspiration and particle type. Results showed that Scindapsus yielded a high PM removal efficiency for smoke type PM2.5 under active transpiration. We examined the correlation of PM removal and relative humidity (RH) and evaluated the increased effect of RH on PM2.5 removal by using a plant-inspired in vitro model. Based on the present results, the increase of RH due to evapotranspiration is crucial to the reduction of PM2.5 using plants.

Enhancement of plant leaf transpiration with effective use of surface acoustic waves: effect of wave frequency.

Water transport in vascular plants provides remarkable opportunities for various engineering applications due to its highly efficient and powerless transportability. Several previous studies were conducted to regulate the biological responses of plants using noninvasive audible or ultrasound waves. However, the control mechanism of acoustic stimuli applied to plants has not been investigated yet. Thus, the practical application of these stimuli to real plants still exhibits technological limitations. This study experimentally investigated the effects of surface acoustic wave (SAW) frequency on plant transpiration to understand the acoustic-activated leaf transpiration and utilize the advantages of SAW. We captured consecutive images of the enhanced water transport in the test plant (Epipremnum aureum) by SAW at three different frequencies (10, 15, and 20 MHz). The dye solution at 15 MHz SAW presented the highest intensity value after 40 min of SAW stimulation. The excitation areas for 15 and 20 MHz SAWs were decreased to 42.3% and 22.6%, respectively, compared with that of 10 MHz SAW. The transpiration rates were directly measured to compare water transport enhancement quantitatively when different SAW frequencies were applied to the same plant leaves. The water transport in the leaves was maximized at 15 MHz SAW, regardless of excitation area.

Hydrodynamic Study on the "Stop-and-Acceleration" Pattern of Refilling Flow at Perforation Plates by Using a Xylem-Inspired Channel.

Porous structures, such as perforation plates and pit membranes, have attracted considerable attention due to their hydraulic regulation of water flow through vascular plant networks. However, limited information is available regarding the hydraulic functions of such structures during water-refilling and embolism repair because of difficulties in simultaneous in vivo measurements of refilling flow and pressure variations in xylem vessels. In this study, we developed a xylem-inspired microchannel with a porous mesh for systematic investigation on the hydraulic contribution of perforation plates on water-refilling. In particular, the "stop-and-acceleration" phenomenon of the water meniscus at the porous mesh structure was carefully examined in macroscopic and microscopic views. This distinctive phenomenon usually occurs in the xylem vessels of vascular plants during embolism repair. Based on the experimental results, we established a theoretical model of the flow characteristics and pressure variations around the porous structure inside the microchannel. Perforation plates could be speculated to be a pressure-modulated flow controller that facilitates embolism recovery. Furthermore, the proposed xylem-inspired channel can be used to investigate the hydraulic functions of porous structures for water management in plants.

Superb feeding behavior of Aedes albopictus transmitting Zika virus.

Disease-mediated mosquitoes have been receiving much attention, as the World Health Organization recently declared the Zika virus a global public health emergency. Mosquitoes transmit pathogens that cause various tropical diseases including malaria, dengue fever and yellow fever as well as Zika virus. The vector efficiency of mosquitoes depends on their blood-feeding characteristics and the mechanics of their blood-sucking pump system, but only a few studies have attempted to investigate these key issues. In this study, we demonstrate the rapid and gluttonous liquid-feeding characteristics of Ae. albopictus which transmits Zika virus can be explained by similar proportion of two blood-sucking pumps and accelerated liquid intake driven by fast expanding of pumps. Our results provide insight into the vector efficiency of Ae. albopictus in terms of feeding velocity, pumping frequency, liquid-intake rate, and wall shear stress.

Nearly Perfect Durable Superhydrophobic Surfaces Fabricated by a Simple One-Step Plasma Treatment.

Fabrication of superhydrophobic surfaces is an area of great interest because it can be applicable to various engineering fields. A simple, safe and inexpensive fabrication process is required to fabricate applicable superhydrophobic surfaces. In this study, we developed a facile fabrication method of nearly perfect superhydrophobic surfaces through plasma treatment with argon and oxygen gases. A polytetrafluoroethylene (PTFE) sheet was selected as a substrate material. We optimized the fabrication parameters to produce superhydrophobic surfaces of superior performance using the Taguchi method. The contact angle of the pristine PTFE surface is approximately 111.0° ± 2.4°, with a sliding angle of 12.3° ± 6.4°. After the plasma treatment, nano-sized spherical tips, which looked like crown-structures, were created. This PTFE sheet exhibits the maximum contact angle of 178.9°, with a sliding angle less than 1°. As a result, this superhydrophobic surface requires a small external force to detach water droplets dripped on the surface. The contact angle of the fabricated superhydrophobic surface is almost retained, even after performing an air-aging test for 80 days and a droplet impacting test for 6 h. This fabrication method can provide superb superhydrophobic surface using simple one-step plasma etching.

Comparison of the tracheal systems of Anopheles sinensis and Aedes togoi larvae using synchrotron X-ray microscopic computed tomography (respiratory system of mosquito larvae using SR-µCT).

Mosquito-borne diseases, such as malaria, dengue fever, and Zika virus, are serious global health issues. Vector control may be an important strategy in reducing the mortality caused by these diseases. The respiratory system of mosquito larvae in the water has to inhale atmospheric oxygen as aquatic organisms. In this study, the three-dimensional (3D) structures of the dorsal longitudinal trunks (DLTs) of the tracheal systems of Anopheles sinensis and Aedes togoi were compared using synchrotron X-ray microscopic computed tomography. DLT respiratory frequencies were also investigated. Interestingly, the larvae of the two mosquito species exhibit tracheal systems that are both morphologically and functionally distinct. A. sinensis hangs horizontally under the water surface, and has a smaller DLT volume than A. togoi. In contrast, A. togoi hangs upside down using a siphon by fixing its tip to the water surface. The frequency of peristaltic movement in A. togoi is higher than that of A. sinensis. These differences in the structures and breathing behaviors of the respiratory systems of mosquito larvae provide new insights into the tracheal systems of mosquito larvae, which should help develop novel effective control strategies targeting mosquito larvae.

Three-dimensional structures of the tracheal systems of Anopheles sinensis and Aedes togoi pupae.

Mosquitoes act as a vector for the transmission of disease. The World Health Organization has recommended strict control of mosquito larvae because of their "few, fixed, and findable" features. The respiratory system of mosquito larvae and pupae in the water has a weak point. As aquatic organisms, mosquito larvae and pupae inhale atmosphere oxygen. However, the mosquito pupae have a non-feeding stage, unlike the larvae. Therefore, detailed study on the tracheal system of mosquito pupae is helpful for understanding their survival strategy. In this study, the three-dimensional (3D) structures of the tracheal systems of Anopheles sinensis and Aedes togoi pupae were comparatively investigated using synchrotron X-ray microscopic computed tomography. The respiratory frequencies of the dorsal trunks were also investigated. Interestingly, the pupae of the two mosquito species possess special tracheal systems of which the morphological and functional features are distinctively different. The respiratory frequency of Ae. togoi is higher than that of An. sinensis. These differences in the breathing phenomena and 3D structures of the respiratory systems of these two mosquito species provide an insight into the tracheal systems of mosquito pupae.

SPECT/CT analysis of splenic function in genistein-treated malaria-infected mice.

Spleen traps malaria-infected red blood cells, thereby leading to splenomegaly. Splenomegaly induces impairment in splenic function, i.e., rupture. Therefore, splenomegaly inhibition is required to protect the spleen. In our previous study, genistein was found to have an influence on malaria-induced splenomegaly. However, the effect of genistein in malaria-induced splenomegaly, especially on the function of spleen, has not been fully investigated. In this study, hematoxylin and eosin (H&E) staining images show that genistein partially prevents malaria-induced architectural disruption of spleen. In addition, genistein decreases transgenic Plasmodium parasites accumulation in the spleen. Genistein treatment can protect splenic function from impairment caused by malaria infection. To examine the functions of malaria-infected spleen, we employed single-photon emission computed tomography/computed tomography (SPECT/CT) technology. Red blood cells are specifically radiolabeled with Technetium-99m pertechnetate (99mTcO4-) and trapped inside the spleen. The standardized uptake values (SUVs) in the spleen of infected mice are higher than those of naive and genistein-treated mice. However, genistein reduces the malaria-induced trapping capacity of spleen for heat-damaged radiolabeled RBCs, while exhibiting a protective effect against malaria. Considering these results, we suggested that genistein could be effectively used in combination therapy for malaria-induced splenic impairment.

In vivo dynamic analysis of water refilling in embolized xylem vessels of intact Zea mays leaves.

Background and Aims The refilling of embolized xylem vessels under tension is a major issue in water transport among vascular plants. However, xylem embolism and refilling remain poorly understood because of technical limitations. Direct observation of embolism repair in intact plants is essential to understand the biophysical aspects of water refilling in embolized xylem vessels. This paper reports on details of the water refilling process in leaves of the intact herbaceous monocot plant Zea mays and its refilling kinetics obtained by a direct visualization technique. Methods A synchrotron X-ray micro-imaging technique was used to monitor water refilling in embolized xylem vessels of intact maize leaves. Xylem embolism was artificially induced by using a glass capillary; real-time images of water refilling dynamics were consecutively captured at a frame rate of 50 f.p.s. Key Results Water supply in the radial direction initiates droplet formation on the wall of embolized xylem vessels. Each droplet grows into a water column; this phenomenon shows translation motion or continuous increase in water column volume. In some instances, water columns merge and form one large water column. Water refilling in the radial direction causes rapid recovery from embolism in several minutes. The average water refilling velocity is approx. 1 µm s-1. Conclusions Non-destructive visualization of embolized xylem vessels demonstrates rapid water refilling and gas bubble removal as key elements of embolism repair in a herbaceous monocot species. The refilling kinetics provides new insights into the dynamic mechanism of water refilling phenomena.

Vulnerability of Protoxylem and Metaxylem Vessels to Embolisms and Radial Refilling in a Vascular Bundle of Maize Leaves.

Regulation of water flow in an interconnected xylem vessel network enables plants to survive despite challenging environment changes that can cause xylem embolism. In this study, vulnerability to embolisms of xylem vessels and their water-refilling patterns in vascular bundles of maize leaves were experimentally investigated by employing synchrotron X-ray micro-imaging technique. A vascular bundle in maize consisted of a protoxylem vessel with helical thickenings between two metaxylem vessels with single perforation plates and nonuniformly distributed pits. When embolism was artificially induced in excised maize leaves by exposing them to air, protoxylem vessels became less vulnerable to dehydration compared to metaxylem vessels. After supplying water into the embolized vascular bundles, when water-refilling process stopped at the perforation plates in metaxylem vessels, discontinuous radial water influx occurred surprisingly in the adjacent protoxylem vessels. Alternating water refilling pattern in protoxylem and metaxylem vessels exhibited probable correlation between the incidence location and time of water refilling and the structural properties of xylem vessels. These results imply that the maintenance of water transport and modulation of water refilling are affected by hydrodynamic roles of perforation plates and radial connectivity in a xylem vascular bundle network.

Direct observation of local xylem embolisms induced by soil drying in intact Zea mays leaves.

The vulnerability of vascular plants to xylem embolism is closely related to their stable long-distance water transport, growth, and survival. Direct measurements of xylem embolism are required to understand what causes embolism and what strategies plants employ against it. In this study, synchrotron X-ray microscopy was used to non-destructively investigate both the anatomical structures of xylem vessels and embolism occurrence in the leaves of intact Zea mays (maize) plants. Xylem embolism was induced by water stress at various soil drying periods and soil water contents. X-ray images of dehydrated maize leaves showed that the ratio of gas-filled vessels to all xylem vessels increased with decreased soil water content and reached approximately 30% under severe water stress. Embolism occurred in some but not all vessels. Embolism in maize leaves was not strongly correlated with xylem diameter but was more likely to occur in the peripheral veins. The rate of embolism formation in metaxylem vessels was higher than in protoxylem vessels. This work has demonstrated that xylem embolism remains low in maize leaves under water stress and that there xylem has characteristic spatial traits of vulnerability to embolism.

Comparison of the functional features of the pump organs of Anopheles sinensis and Aedes togoi.

Mosquitoes act as vectors for severe tropical diseases. Mosquito-borne diseases are affected by various factors such as environmental conditions, host body susceptibility, and mosquito feeding behavior. Among these factors, feeding behavior is affected by the feeding pump system located inside the mosquito head and also depends on the species of mosquito. Therefore, the 3D morphological structures of the feeding pumps of Aedes togoi and Anopheles sinensis were comparatively investigated using synchrotron X-ray microscopic computed tomography. In addition, the feeding behaviors of their pumping organs were also investigated using a 2D X-ray micro-imaging technique. An. sinensis, a malarial vector mosquito, had a larger feeding pump volume than Ae. togoi in the static or resting position. Interestingly, the two species of mosquitoes exhibited different feeding behaviors. Ae. togoi had a higher feeding frequency and expansion ratio than An. sinensis. Ae. togoi also exhibited F-actin localization more clearly. These distinctive variations in feeding volumes and behaviors provide essential insight into the blood-feeding mechanisms of female mosquitoes as vectors for tropical diseases.

Functional water flow pathways and hydraulic regulation in the xylem network of Arabidopsis.

In vascular plants, the xylem network constitutes a complex microfluidic system. The relationship between vascular network architecture and functional hydraulic regulation during actual water flow remains unexplored. Here, we developed a method to visualize individual xylem vessels of the 3D xylem network of Arabidopsis thaliana, and to analyze the functional activities of these vessels using synchrotron X-ray computed tomography with hydrophilic gold nanoparticles as flow tracers. We show how the organization of the xylem network changes dynamically throughout the plant, and reveal how the elementary units of this transport system are organized to ensure both long-distance axial water transport and local lateral water transport. Xylem vessels form distinct clusters that operate as functional units, and the activity of these units, which determines water flow pathways, is modulated not only by varying the number and size of xylem vessels, but also by altering their interconnectivity and spatial arrangement. Based on these findings, we propose a regulatory model of water transport that ensures hydraulic efficiency and safety.

In vivo monitoring of intracellular chloroplast movements in intact leaves of C4 plants using two-photon microscopy.

Dynamic changes in the spatial distribution of chloroplasts are essential for optimizing photosynthetic capacity under changing light conditions. Light-induced movement of chloroplasts has been widely investigated, but most studies were conducted on isolated tissues or protoplasts. In this study, a two-photon microscopy (TPM) system was adapted to monitor the intracellular 3-dimensional (3D) movements of chloroplasts in intact leaves of plants during dark to light transitions. The TPM imaging was based on autofluorescence of chlorophyll generated by a femto-second Ti:Sapphire laser. All chloroplasts did not exhibit the same motion in response to irradiation variation. In the sub-epidermal mesophyll cells, chloroplasts generally moved away from the surface following blue light treatment, however many chloroplasts did not show any movement. Such spatial heterogeneity in chloroplast motility underlines the importance of monitoring intracellular orientation and movement of individual chloroplasts across intact leaves. Our investigation shows that the 3D imaging of chloroplasts using TPM can help to understand the changes in local photosynthetic capacity in intact leaves under changing environmental conditions.

Interactive ion-mediated sap flow regulation in olive and laurel stems: physicochemical characteristics of water transport via the pit structure.

Sap water is distributed and utilized through xylem conduits, which are vascular networks of inert pipes important for plant survival. Interestingly, plants can actively regulate water transport using ion-mediated responses and adapt to environmental changes. However, ionic effects on active water transport in vascular plants remain unclear. In this report, the interactive ionic effects on sap transport were systematically investigated for the first time by visualizing the uptake process of ionic solutions of different ion compositions (K+/Ca2+) using synchrotron X-ray and neutron imaging techniques. Ionic solutions with lower K+/Ca2+ ratios induced an increased sap flow rate in stems of Olea europaea L. and Laurus nobilis L. The different ascent rates of ionic solutions depending on K+/Ca2+ ratios at a fixed total concentration increases our understanding of ion-responsiveness in plants from a physicochemical standpoint. Based on these results, effective structural changes in the pit membrane were observed using varying ionic ratios of K+/Ca2+. The formation of electrostatically induced hydrodynamic layers and the ion-responsiveness of hydrogel structures based on Hofmeister series increase our understanding of the mechanism of ion-mediated sap flow control in plants.

Label-free viscosity measurement of complex fluids using reversal flow switching manipulation in a microfluidic channel.

The accurate viscosity measurement of complex fluids is essential for characterizing fluidic behaviors in blood vessels and in microfluidic channels of lab-on-a-chip devices. A microfluidic platform that accurately identifies biophysical properties of blood can be used as a promising tool for the early detections of cardiovascular and microcirculation diseases. In this study, a flow-switching phenomenon depending on hydrodynamic balancing in a microfluidic channel was adopted to conduct viscosity measurement of complex fluids with label-free operation. A microfluidic device for demonstrating this proposed method was designed to have two inlets for supplying the test and reference fluids, two side channels in parallel, and a junction channel connected to the midpoint of the two side channels. According to this proposed method, viscosities of various fluids with different phases (aqueous, oil, and blood) in relation to that of reference fluid were accurately determined by measuring the switching flow-rate ratio between the test and reference fluids, when a reverse flow of the test or reference fluid occurs in the junction channel. An analytical viscosity formula was derived to measure the viscosity of a test fluid in relation to that of the corresponding reference fluid using a discrete circuit model for the microfluidic device. The experimental analysis for evaluating the effects of various parameters on the performance of the proposed method revealed that the fluidic resistance ratio ( R J L / R L , fluidic resistance in the junction channel ( R J L ) to fluidic resistance in the side channel ( R L )) strongly affects the measurement accuracy. The microfluidic device with smaller R J L / R L values is helpful to measure accurately the viscosity of the test fluid. The proposed method accurately measured the viscosities of various fluids, including single-phase (Glycerin and plasma) and oil-water phase (oil vs. deionized water) fluids, compared with conventional methods. The proposed method was also successfully applied to measure viscosities of blood with varying hematocrits, chemically fixed RBCS, and channel sizes. Based on these experimental results, the proposed method can be effectively used to measure the viscosities of various fluids easily, without any fluorescent labeling and tedious calibration procedures.

Development and clinical evaluation of a rapid serodiagnostic test for toxoplasmosis of cats using recombinant SAG1 antigen.

Rapid serodiagnostic methods for Toxoplasma gondii infection in cats are urgently needed for effective control of transmission routes toward human infections. In this work, 4 recombinant T. gondii antigens (SAG1, SAG2, GRA3, and GRA6) were produced and tested for the development of rapid diagnostic test (RDT). The proteins were expressed in Escherichia coli, affinity-purified, and applied onto the nitrocellulose membrane of the test strip. The recombinant SAG1 (rSAG1) showed the strongest antigenic activity and highest specificity among them. We also performed clinical evaluation of the rSAG1-loaded RDT in 182 cat sera (55 household and 127 stray cats). The kit showed 0.88 of kappa value comparing with a commercialized ELISA kit, which indicated a significant correlation between rSAG1-loaded RDT and the ELISA kit. The overall sensitivity and specificity of the RDT were 100% (23/23) and 99.4% (158/159), respectively. The rSAG1-loaded RDT is rapid, easy to use, and highly accurate. Thus, it would be a suitable diagnostic tool for rapid detection of antibodies in T. gondii-infected cats under field conditions.