Capillarity-Driven Enrichment and Hydrodynamic Trapping of Trace Nucleic Acids by Plasmonic Cavity Membrane for Rapid and Sensitive Detections.

Small-reactor-based polymerase chain reaction (PCR) has attracted considerable attention. A significant number of tiny reactors must be prepared in parallel to capture, amplify, and accurately quantify few target genes in clinically relevant large volume, which, however, requires sophisticated microfabrication and longer sample-to-answer time. Here, single plasmonic cavity membrane is reported that not only enriches and captures few nucleic acids by taking advantage of both capillarity and hydrodynamic trapping but also quickly amplifies them for sensitive plasmonic detection. The plasmonic cavity membrane with few nanoliters in a void volume is fabricated by self-assembling gold nanorods with SiO2 tips. Simulations reveal that hydrodynamic stagnation between the SiO2 tips is mainly responsible for the trapping of the nucleic acid in the membrane. Finally, it is shown that the plasmonic cavity membrane is capable of enriching severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genes up to 20 000-fold within 1 min, amplifying within 3 min, and detecting the trace genes as low as a single copy µL-1. It is anticipated that this work not only expands the utility of PCR but also provides an innovative way of the enrichment and detection of trace biomolecules in a variety of point-of-care testing applications.

Electron-Transport-Chain-Mediated Selective Growth of Gold Nanocrystals in the Intermembrane Space of Live Microbial Cells.

Hybridization of microbial cells with inorganic nanoparticles that could dramatically improve cellular functions such as electron transfer has been realized by the random attachment or stochastic entry of the nanoparticles. Clearly, the selective growth of inorganic nanoparticles on target functional organelles is ideal for such hybridization. Here, we report the selective growth of gold nanocrystals in the intermembrane space (IMS) of Escherichia coli by exploiting the electron transport chain (ETC). We systematically show that gold ions are permeated through porins in the outer membrane of E. coli and further reduced to gold nanocrystals by the ETC in live E. coli. We directly observe that the resulting gold nanocrystals exist only in the IMS by transmission electron microscopy measurements of cross-sectioned E. coli. Molecular dynamics simulations suggest that once gold ions are reduced to small nuclei by the ETC, the nuclei can be stably physisorbed onto ETC complexes, further supporting the ETC-mediated growth. Finally, we show that the ATP synthesis of E. coli where gold nanocrystals are formed in the IMS is up to 9 times higher than that of E. coli alone. We believe that our work can significantly contribute to not only improving microbial metabolic functions for biological energy conversion but also restoring physiological dysfunctions of microbial cells for biomedicine.

Bubble-free diatoms polymerase chain reaction.

Polymerase chain reaction (PCR) in small fluidic systems not only improves speed and sensitivity of deoxyribonucleic acid (DNA) amplification but also achieves high-throughput quantitative analyses. However, air bubble trapping and growth during PCR has been considered as a critical problem since it causes the failure of DNA amplification. Here we report bubble-free diatom PCR by exploiting a hierarchically porous silica structure of single-celled algae. We show that femtoliters of PCR solution can be spontaneously loaded into the diatom interior without air bubble trapping due to the surface hydrophilicity and pore structure of the diatom. We discover that a large pressure gradient between air bubbles and nanopores rapidly removes residual air bubbles through the periodically arrayed nanopores during thermal cycling. We demonstrate the DNA amplification by diatom PCR without air bubble trapping and growth. Finally, we successfully detect DNA fragments of SARS-CoV-2 with as low as 10 copies/µl by devising a microfluidic device integrated with diatoms assembly. We believe that our work can be applied to many PCR applications for innovative molecular diagnostics and provides new opportunities for naturally abundant diatoms to create innovative biomaterials in real-world applications.

Elucidation of Phytochemicals Affecting Platelet Responsiveness in Dangguisu-san: Active Ingredient Prediction and Experimental Research Using Network Pharmacology.

Plant-derived phytochemicals are emerging as novel agents for protection against chronic disorders. Dangguisu-san is a herbal prescription to invigorate the blood and relieve pain. Among the numerous active constituents of Dangguisu-san, those expected to be effective at inhibiting platelet aggregation were predicted using a network pharmacological method, and their efficacy was experimentally demonstrated. All four identified chemical components, namely chrysoeriol, apigenin, luteolin, and sappanchalcone, suppressed the aggregation of platelets to a certain extent. However, we report, for the first time, that chrysoeriol acts as a strong inhibitor of platelet aggregation. Although additional in vivo studies are needed, among the complex constituents of herbal medicines, the components that exert an inhibitory effect on platelet aggregation were predicted using a network pharmacological method and experimentally confirmed with human platelets.

Inner-Membrane-Bound Gold Nanoparticles as Efficient Electron Transfer Mediators for Enhanced Mitochondrial Electron Transport Chain Activity.

Electron transfer through the mitochondrial electron transport chain (ETC) can be critically blocked by the dysfunction of protein complexes. Redox-active molecules have been used to mediate the electron transfer in place of the dysfunctional complexes; however, they are limited to replacing complex I and are known to be toxic. Here we report artificial mitochondrial electron transfer pathways that enhance ETC activity by exploiting inner-membrane-bound gold nanoparticles (GNPs) as efficient electron transfer mediators. The hybridization of mitochondria with GNPs, driven by electrostatic interaction, is successfully visualized in real time at the level of a single mitochondrion. By observing quantized quenching dips via plasmon resonance energy transfer, we reveal that the hybridized GNPs are bound to the inner membrane of mitochondria irrespective of the presence of the outer membrane. The ETC activity of mitochondria with GNPs such as membrane potential, oxygen consumption, and ATP production is remarkably increased in vitro.

Sensitive and Direct Optical Monitoring of Release and Cellular Uptake of Aqueous CO from CO-Releasing Molecules.

Dynamics of release and cellular uptake of aqueous CO from CO-releasing molecules (CORMs) significantly affect signaling and cell viability. So far, it has been mainly observed by IR, UV-visible, and fluorescence techniques, which suffer from poor sensitivity and slow response time. Here, we show how to directly probe the mass transfer of aqueous CO from CORMs to cells using a fluidic chamber integrated with live cells and Raman reporters of large-area Au@Pd core-shell nanoparticle assembly to emulate a physiologically relevant microenvironment. We sensitively and directly detect CO release from trace CORMs of as low as 100 nM by measuring the Raman transitions of CO via rapid chemisorption onto the surface of the Au@Pd nanoparticles. By using our method, we successfully observe the dynamics of CO release from CORM-2 despite its very short half-life. We also reveal that the initial rate of CO release from CORM-3 is dramatically decreased by tens to hundreds of times when exposed to physiologically relevant pH variations from 7.4 to 2.5, which can be attributed to the acid hydrolysis of the CO ligand. CORM-2 tends to quickly release CO regardless of pH, probably because of its rapid cleavage into two monomeric Ru complexes by the co-solvent. The decrease in the initial rate at lower temperatures is more significant for CORM-3 than for CORM-2. Finally, we observe that the cellular uptake of aqueous CO from CORM-3 by lung cancer cells is approximately 2 times higher than that of normal lung cells.

Liposome/gold hybrid nanoparticle encoded with CoQ10 (LGNP-CoQ10) suppressed rheumatoid arthritis via STAT3/Th17 targeting.

Coenzyme Q10 (CoQ10), also known as ubiquinone, is a fat-soluble antioxidant. Although CoQ10 has not been approved as medication by the Food and Drug Administration, it is widely used in dietary supplements. Some studies have shown that CoQ10 has anti-inflammatory effects on various autoimmune disorders. In this study, we investigated the anti-inflammatory effects of liposome/gold hybrid nanoparticles encoded with CoQ10 (LGNP-CoQ10). Both CoQ10 and LGNP-CoQ10 were administered orally to mice with collagen-induced arthritis (CIA) for 10 weeks. The inflammation pathology of joint tissues of CIA mice was then analyzed using hematoxylin and eosin and Safranin O staining, as well as immunohistochemistry analysis. We obtained immunofluorescence staining images of spleen tissues using confocal microscopy. We found that pro-inflammatory cytokines were significantly decreased in LGNP-CoQ10 injected mice. Th17 cell and phosphorylated STAT3-expressed cell populations were also decreased in LGNP-CoQ10 injected mice. When human peripheral blood mononuclear cells (PBMCs) were treated with CoQ10 and LGNP-CoQ10, the IL-17 expression of PBMCs in the LGNP-CoQ10-treated group was significantly reduced. Together, these results suggest that LGNP-CoQ10 has therapeutic potential for the treatment of rheumatoid arthritis.

Direct Optical and Ultrasensitive Probing of Nonequilibrium Dynamics of Carbon Monoxide in an Aqueous Phase during Biochemical Reactions.

Detection of trace carbon monoxide (CO) dissolved in an aqueous phase is key for monitoring and optimizing biological and chemical gas conversions. So far, irrespective of the nonequilibrium nature of these conversion processes, because of low water solubility of CO, such detection has been performed indirectly, under the assumption of thermodynamic equilibrium, by the combination of chromatographic measurement of relatively abundant CO in a gas phase and Henry's law. Direct and sensitive detection of dissolved CO under nonequilibrium has not been explored yet. Here, we report the direct, ultrasensitive, and real-time monitoring of nonequilibrium dynamics of CO in an aqueous phase during biochemical conversions by devising miniaturized fluidic reactors with built-in CO-specific optical probes via surface-enhanced Raman spectroscopy. As the sensitive and selective probes, we fabricate ligand-free Au@Pd core-shell nanoparticle monolayers to maximize the Raman signal of single CO in the aqueous phase. We confirm that under equilibrium conditions, aqueous and gaseous CO concentrations estimated by our method are in good agreement with those measured directly and indirectly by gas chromatography (GC). We show that our probe can detect the aqueous CO concentrations as low as ca. 0.01% with high signal reproducibility, which is 200-fold more sensitive than that achieved by infrared spectroscopy. Finally, we successfully observe the nonequilibrium dynamics of the aqueous CO during biochemical reactions, which cannot be sensed by other detection methods including even indirect measurement by GC. We anticipate that our method can be widely applied not only for monitoring of biochemical gas reactions on multiple scales from a large reactor to a single-molecule level but also for molecular imaging of biological systems.

Plasmonic bacteria on a nanoporous mirror via hydrodynamic trapping for rapid identification of waterborne pathogens.

A rapid, precise method for identifying waterborne pathogens is critically needed for effective disinfection and better treatment. However, conventional methods, such as culture-based counting, generally suffer from slow detection times and low sensitivities. Here, we developed a rapid detection method for tracing waterborne pathogens by an innovative optofluidic platform, a plasmonic bacteria on a nanoporous mirror, that allows effective hydrodynamic cell trapping, enrichment of pathogens, and optical signal amplifications. We designed and simulated the integrated optofluidic platform to maximize the enrichment of the bacteria and to align bacteria on the nanopores and plasmonic mirror via hydrodynamic cell trapping. Gold nanoparticles are self-assembled to form antenna arrays on the surface of bacteria, such as Escherichia coli and Pseudomonas aeruginosa, by replacing citrate with hydroxylamine hydrochloride in order to amplify the signal of the plasmonic optical array. Owing to the synergistic contributions of focused light via the nanopore geometry, self-assembled nanoplasmonic optical antennas on the surface of bacteria, and plasmonic mirror, we obtain a sensitivity of detecting E. coli as low as 102 cells/ml via surface-enhanced Raman spectroscopy. We believe that our label-free strategy via an integrated optofluidic platform will pave the way for the rapid, precise identification of various pathogens.

Tunable Plasmonic Cavity for Label-free Detection of Small Molecules.

Owing to its high sensitivity and high selectivity along with rapid response time, plasmonic detection has gained considerable interest in a wide variety of sensing applications. To improve the fieldwork applicability and reliability of plasmonic detection, the integration of plasmonic nanoparticles into optical devices is desirable. Herein, we propose an integrated label-free detection platform comprising a plasmonic cavity that allows sensitive molecular detection via either surface-enhanced Raman scattering (SERS) or plasmon resonance energy transfer (PRET). A small droplet of metal ion solution spontaneously produces a plasmonic cavity on the surface of uncured poly(dimethylsiloxane) (PDMS), and as PDMS is cured, the metal ions are reduced to form a plasmonic antennae array on the cavity surface. Unique spherical feature and the integrated metallic nanoparticles of the cavity provide excellent optical functions to focus the incident light in the cavity and to rescatter the light absorbed by the nanoparticles. The optical properties of the plasmonic cavity for SERS or PRET are optimized by controlling the composition, size, and density of the metal nanoparticles. By using the cavity, we accomplish both 1000-fold sensitive detection and real-time monitoring of reactive oxygen species secreted by live cells via PRET. In addition, we achieve sensitive detection of trace amounts of toxic environmental molecules such as 5-chloro-2-methyl-4-isothiazolin-3-one/2-methyl-4-isothiazol-3-one (CMIT/MIT) and bisphenol A, as well as several small biomolecules such as glucose, adenine, and tryptophan, via SERS.

General and programmable synthesis of hybrid liposome/metal nanoparticles.

Hybrid liposome/metal nanoparticles are promising candidate materials for biomedical applications. However, the poor selectivity and low yield of the desired hybrid during synthesis pose a challenge. We designed a programmable liposome by selective encoding of a reducing agent, which allows self-crystallization of metal nanoparticles within the liposome to produce stable liposome/metal nanoparticles alone. We synthesized seven types of liposome/monometallic and more complex liposome/bimetallic hybrids. The resulting nanoparticles are tunable in size and metal composition, and their surface plasmon resonance bands are controllable in visible and near infrared. Owing to outer lipid bilayer, our liposome/Au nanoparticle shows better colloidal stability in biologically relevant solutions as well as higher endocytosis efficiency than gold nanoparticles without the liposome. We used this hybrid in intracellular imaging of living cells via surface-enhanced Raman spectroscopy, taking advantage of its improved physicochemical properties. We believe that our method greatly increases the utility of metal nanoparticles in in vivo applications.

Spontaneous Self-Formation of 3D Plasmonic Optical Structures.

Self-formation of colloidal oil droplets in water or water droplets in oil not only has been regarded as fascinating fundamental science but also has been utilized in an enormous number of applications in everyday life. However, the creation of three-dimensional (3D) architectures by a liquid droplet and an immiscible liquid interface has been less investigated than other applications. Here, we report interfacial energy-driven spontaneous self-formation of a 3D plasmonic optical structure at room temperature without an external force. Based on the densities and interfacial energies of two liquids, we simulated the spontaneous formation of a plasmonic optical structure when a water droplet containing metal ions meets an immiscible liquid polydimethylsiloxane (PDMS) interface. At the interface, the metal ions in the droplet are automatically reduced to form an interfacial plasmonic layer as the liquid PDMS cures. The self-formation of both an optical cavity and integrated plasmonic nanostructure significantly enhances the fluorescence by a magnitude of 1000. Our findings will have a huge impact on the development of various photonic and plasmonic materials as well as metamaterials and devices.

Consequence of climate mitigation on the risk of hunger.

Climate change and mitigation measures have three major impacts on food consumption and the risk of hunger: (1) changes in crop yields caused by climate change; (2) competition for land between food crops and energy crops driven by the use of bioenergy; and (3) costs associated with mitigation measures taken to meet an emissions reduction target that keeps the global average temperature increase to 2 °C. In this study, we combined a global computable general equilibrium model and a crop model (M-GAEZ), and we quantified the three impacts on risk of hunger through 2050 based on the uncertainty range associated with 12 climate models and one economic and demographic scenario. The strong mitigation measures aimed at attaining the 2 °C target reduce the negative effects of climate change on yields but have large negative impacts on the risk of hunger due to mitigation costs in the low-income countries. We also found that in a strongly carbon-constrained world, the change in food consumption resulting from mitigation measures depends more strongly on the change in incomes than the change in food prices.

Interfacial Synthesis of Two-Dimensional Dendritic Platinum Nanoparticles Using Oleic Acid-in-Water Emulsion.

Here we propose facile and scalable synthesis of two-dimensional (2D) dendritic platinum nanoparticle at room temperature by exploiting an oil-in-water emulsion. The interfacial synthesis selectively provides platinum nanoparticle with 2D structure in high yield by controlling key reactants such as the amount of oleic acid and the concentration of block copolymer. Electrocatalytic activity of 2D dendritic platinum nanoparticle for oxygen reduction and methanol oxidation reaction is also examined.

Two-dimensional hyper-branched gold nanoparticles synthesized on a two-dimensional oil/water interface.

Two-dimensional (2D) gold nanoparticles can possess novel physical and chemical properties, which will greatly expand the utility of gold nanoparticles in a wide variety of applications ranging from catalysis to biomedicine. However, colloidal synthesis of such particles generally requires sophisticated synthetic techniques to carefully guide anisotropic growth. Here we report that 2D hyper-branched gold nanoparticles in the lateral size range of about 50 ~ 120 nm can be synthesized selectively on a 2D immiscible oil/water interface in a few minutes at room temperature without structure-directing agents. An oleic acid/water interface can provide diffusion-controlled growth conditions, leading to the structural evolution of a smaller gold nucleus to 2D nanodendrimer and nanourchin at the interface. Simulations based on the phase field crystal model match well with experimental observations on the 2D branching of the nucleus, which occurs at the early stage of growth. Branching results in higher surface area and stronger near-field enhancement of 2D gold nanoparticles. This interfacial synthesis can be scaled up by creating an emulsion and the recovery of oleic acid is also achievable by centrifugation.

Climate change impact and adaptation assessment on food consumption utilizing a new scenario framework.

We assessed the impacts of climate change and agricultural autonomous adaptation measures (changes in crop variety and planting dates) on food consumption and risk of hunger considering uncertainties in socioeconomic and climate conditions by using a new scenario framework. We combined a global computable general equilibrium model and a crop model (M-GAEZ), and estimated the impacts through 2050 based on future assumptions of socioeconomic and climate conditions. We used three Shared Socioeconomic Pathways as future population and gross domestic products, four Representative Concentration Pathways as a greenhouse gas emissions constraint, and eight General Circulation Models to estimate climate conditions. We found that (i) the adaptation measures are expected to significantly lower the risk of hunger resulting from climate change under various socioeconomic and climate conditions. (ii) population and economic development had a greater impact than climate conditions for risk of hunger at least throughout 2050, but climate change was projected to have notable impacts, even in the strong emission mitigation scenarios. (iii) The impact on hunger risk varied across regions because levels of calorie intake, climate change impacts and land scarcity varied by region.