Rapid Volumetric Optoacoustic Tracking of Individual Microparticles In Vivo Enabled by a NIR-Absorbing Gold-Carbon Shell.

Rapid volumetric in vivo visualization of circulating microparticles can facilitate new biomedical applications, such as blood flow characterization or targeted drug delivery. However, existing imaging modalities generally lack the sensitivity to detect the weak signals generated by individual micrometer-sized particles distributed across millimeter- to centimeter-scale depths in living mammalian tissues. Also, the temporal resolution is typically insufficient to track the particles in an entire three-dimensional region. Herein, we introduce a new type of monodisperse (4 µm) silica-core microparticle coated with a shell formed by a multilayered structure of carbon nanotubes (CNT) and gold nanoparticles (AuNP) to provide strong optoacoustic (OA) absorption-based contrast. We capitalize on the unique advantages of a state-of-the-art high-frame-rate OA tomography system to visualize and track the motion of these core-shell particles individually and volumetrically as they flow throughout the mouse brain vasculature. The feasibility of localizing individual solid particles smaller than red blood cells opens new opportunities for mapping the blood flow velocity, enhancing the resolution and visibility of OA images, and developing new biosensing assays.

Logic-signal-based multiplex detection of MiRNAs with high tension hybridization and multiple signal amplification.

MicroRNAs (miRNAs) are crucial regulators of gene expression. The abnormal expression of miRNA is often closely related to many diseases. However, the accurate clinical profiling of miRNA expression remains a great challenge due to the high similarity and wide variety of miRNA sequence structures. Here, we report a highly specific and sensitive multiplex miRNA detection scheme with high tension hybridization and dual signal amplification based on the recyclable autocatalytic DNAzyme and a light harvesting conjugated polymer. Multiple signals can be read out simultaneously by single excitation through the efficient multiple fluorescence resonance energy transfer (FRET) between the conjugated polymer and different small molecule dyes. In addition, different types of logic gates can also be operated by observing the emission intensities of the labeling dyes with miRNAs as inputs, thus giving rise to a new way for the specific detection of certain miRNAs according to the logic signals. Furthermore, we successfully applied the strategy for multiple miRNA detection in cell lysates and the results agree well with those of qRT-PCR. Thus, we believe that this platform holds great potential for miRNA detection in biological samples.

Dye-Loaded Quatsomes Exhibiting FRET as Nanoprobes for Bioimaging.

Fluorescent organic nanoparticles (FONs) are emerging as an attractive alternative to the well-established fluorescent inorganic nanoparticles or small organic dyes. Their proper design allows one to obtain biocompatible probes with superior brightness and high photostability, although usually affected by low colloidal stability. Herein, we present a type of FONs with outstanding photophysical and physicochemical properties in-line with the stringent requirements for biomedical applications. These FONs are based on quatsome (QS) nanovesicles containing a pair of fluorescent carbocyanine molecules that give rise to Förster resonance energy transfer (FRET). Structural homogeneity, high brightness, photostability, and high FRET efficiency make these FONs a promising class of optical bioprobes. Loaded QSs have been used for in vitro bioimaging, demonstrating the nanovesicle membrane integrity after cell internalization, and the possibility to monitor the intracellular vesicle fate. Taken together, the proposed QSs loaded with a FRET pair constitute a promising platform for bioimaging and theranostics.

Radiation Force as a Physical Mechanism for Ultrasonic Neurostimulation of the Ex Vivo Retina.

Focused ultrasound has been shown to be effective at stimulating neurons in many animal models, both in vivo and ex vivo Ultrasonic neuromodulation is the only noninvasive method of stimulation that could reach deep in the brain with high spatial-temporal resolution, and thus has potential for use in clinical applications and basic studies of the nervous system. Understanding the physical mechanism by which energy in a high acoustic frequency wave is delivered to stimulate neurons will be important to optimize this technology. We imaged the isolated salamander retina of either sex during ultrasonic stimuli that drive ganglion cell activity and observed micron scale displacements, consistent with radiation force, the nonlinear delivery of momentum by a propagating wave. We recorded ganglion cell spiking activity and changed the acoustic carrier frequency across a broad range (0.5-43 MHz), finding that increased stimulation occurs at higher acoustic frequencies, ruling out cavitation as an alternative possible mechanism. A quantitative radiation force model can explain retinal responses and could potentially explain previous in vivo results in the mouse, suggesting a new hypothesis to be tested in vivo Finally, we found that neural activity was strongly modulated by the distance between the transducer and the electrode array showing the influence of standing waves on the response. We conclude that radiation force is the dominant physical mechanism underlying ultrasonic neurostimulation in the ex vivo retina and propose that the control of standing waves is a new potential method to modulate these effects.SIGNIFICANCE STATEMENT Ultrasonic neurostimulation is a promising noninvasive technology that has potential for both basic research and clinical applications. The mechanisms of ultrasonic neurostimulation are unknown, making it difficult to optimize in any given application. We studied the physical mechanism by which ultrasound is converted into an effective energy form to cause neurostimulation in the retina and find that ultrasound acts via radiation force leading to a mechanical displacement of tissue. We further show that standing waves have a strong modulatory effect on activity. Our quantitative model by which ultrasound generates radiation force and leads to neural activity will be important in optimizing ultrasonic neurostimulation across a wide range of applications.

Novel Light-Responsive Hydrogels with Antimicrobial and Antifouling Capabilities.

Smart materials with both bactericidal and bacteria-resistant functions are promising for combating the infection concern of medical devices. Current work mostly utilizes hydrolysis to switch materials from antimicrobial to antifouling forms by incubating materials in aqueous solutions for hours to days. In this work, a new photoresponsive poly[2-((4,5-dimethoxy-2-nitrobenzyl)oxy)- N-(2-(methacryloyloxy)ethyl)- N, N-dimethyl-2-oxoethan-1-aminium] (polyCBNA) hydrogel was developed, incorporating the photolabile 4,5-dimethoxy-2-nitrobenzyl and cationic quaternary ammonium groups. The photolabile groups were readily cleaved from the hydrogel shortly upon UV irradiation at 365 nm (a long wavelength widely used for biomedical applications), leading to polymer surface charge switching from cationic to zwitterionic form. Protein adsorbed significantly on polyCBNA but easily desorbed from surfaces after UV irradiation. The cationic hydrogel as a precursor was shown to effectively kill the attached bacteria, and then quickly switched to zwitterionic antifouling form via photolysis, which released the attached bacteria from surfaces and prevented further bacterial attachment. Moreover, the adhered endothelial cells were easily detached from polyCBNA surfaces triggered by light, providing a facile and less destructive nonenzymatic approach to harvest cells. This smart photoresponsive polyCBNA polymer, with integrated antimicrobial and antifouling properties, holds great potential in biomedical applications such as self-sterilizing and self-cleaning coatings for implants, cell harvesting, and cell patterning.

Facile surface functionalization of upconversion nanoparticles with phosphoryl pillar[5]arenes for controlled cargo release and cell imaging.

Upon surface functionalization and stabilization of ß-NaYF4:Yb/Er upconversion nanoparticles (UCNPs) with phosphoryl pillar[5]arenes (PP5) via a facile ligand exchange method, we constructed a new hybrid material (PP5-UCNPs) with good water dispersibility especially in a physiological environment and superior capability of controlled cargo release and cell imaging.

Oxidative degradation of tetramethylammonium hydroxide (TMAH) by UV/persulfate and associated acute toxicity assessment.

Tetramethylammonium hydroxide (TMAH) is widely used in high-tech industries as a developing agent. Ultraviolet (UV) light-activated persulfate (PS, S2O82-) can be used to generate strongly oxidative sulfate radicals, and it also exhibits the potential to treat TMAH-containing wastewater. This study initially investigated the effect of S2O82- concentration and UV strength on the UV/S2O82- process for the degradation of TMAH in a batch reactor. The results suggested that 15 watts (W) of UV-activated S2O82- at concentrations of 10 or 50 mM resulted in pseudo-first-order TMAH degradation rate constants of 3.1-4.2 × 10-2 min-1, which was adopted for determining the hydraulic retention time (HRT) in a continuous stirred tank reactor (CSTR). The operating conditions (15 W UV/10 mM S2O82-) with a HRT of 129 min resulted in stable residual concentrations of S2O82- and TMAH at approximately 2.6 mM and 20 mg L-1 in effluent, respectively. Several TMAH degradation intermediates including trimethylamine, dimethylamine, and methylamine were also detected. The effluent was adjusted to a neutral pH and evaluated for its biological acute toxicity using Cyprinus carpio as a bioassay organism. The "bio-acute toxicity unit" (TUa) was determined to be 1.41, which indicated that the effluent was acceptable for being discharged into an aquatic ecosystem.

Facile synthesis of TiO2 nanocrystals using NH4F as morphology-controlling agent and its influences on photocatalytic activity.

Controllable synthesis of nanomaterials with different morphologies can significantly affect their properties. Here we report that the morphology and facet orientation of TiO2 nanocrystals can be readily modulated via a hydrothermal method using a simple morphology-controlling agent of NH4F. The photocatalytic activity of resultant TiO2 has been evaluated by photodegradation of methyl orange. The results indicate that the introduction of NH4F can be used to modulate the mophology and, thereby, the photocatalytic activity of TiO2. The obtained TiO2 with high-energy facet, small size, and large surface area can exhibit an improved photocatalytic efficiency, which may be promising for real application.

A 1H NMR assay for measuring the photostationary States of photoswitchable ligands.

Incorporation of photoisomerizable chromophores into small molecule ligands represents a general approach for reversibly controlling protein function with light. Illumination at different wavelengths produces photostationary states (PSSs) consisting of different ratios of photoisomers. Thus optimal implementation of photoswitchable ligands requires knowledge of their wavelength sensitivity. Using an azobenzene-based ion channel blocker as an example, this protocol describes a (1)H NMR assay that can be used to precisely determine the isomeric content of photostationary states (PSSs) as a function of illumination wavelength. Samples of the photoswitchable ligand are dissolved in deuterated water and analyzed by UV/VIS spectroscopy to identify the range of illumination wavelengths that produce PSSs. The PSSs produced by these wavelengths are quantified using (1)H NMR spectroscopy under continuous irradiation through a monochromator-coupled fiber-optic cable. Because aromatic protons of azobenzene trans and cis isomers exhibit sufficiently different chemical shifts, their relative abundances at each PSS can be readily determined by peak integration. Constant illumination during spectrum acquisition is essential to accurately determine PSSs from molecules that thermally relax on the timescale of minutes or faster. This general protocol can be readily applied to any photoswitch that exhibits distinct (1)H NMR signals in each photoisomeric state.

Titania/CnTAB Nanoskeleton as adsorbent and photocatalyst for removal of alkylphenols dissolved in water.

We report here on the removal of alkylphenols (phenol, 4-n-propylphenol, 4-n-heptylphenol and 4-nonylphenol) dissolved in water using the composite particles of nanocrystalline titania and alkyltrimethylammonium bromide (CnH2n+1N(CH3)3Br, CnTAB; n=12, 14, 16 and 18) (named as TiO2/CnTAB Nanoskeleton) as adsorbents and photocatalysts. In particular, the adsorption of alkylphenols onto TiO2/CnTAB Nanoskeleton in water was investigated in terms of hydrophobic interaction between alkylphenols and CnTAB, surface area, pore structure and crystal size of TiO2/CnTAB Nanoskeleton. We revealed that CnTAB incorporated in the TiO2/CnTAB Nanoskeleton promotes the adsorption of alkylphenols onto TiO2/CnTAB Nanoskeleton due to the hydrophobic interaction between alkylphenols and CnTAB. On the other hand, the surface area, pore structure and crystal size of TiO2/CnTAB Nanoskeleton did not affect the adsorption of alkylphenols onto TiO2/CnTAB Nanoskeleton. We also found that the alkylphenols dissolved in water were completely removed by the combination of adsorption and photocatalytic degradation by the TiO2/CnTAB Nanoskeleton under UV irradiation. These results prove that the TiO2/CnTAB Nanoskeleton acts as in tandem an adsorbent and a photocatalyst for removal of alkylphenols dissolved in water.

Characterization and detoxification of a mature landfill leachate using a combined coagulation-flocculation/photo Fenton treatment.

The aim of the present work was to characterize and treat a mature landfill leachate using a coagulation/flocculation process followed by a photo-Fenton oxidation treatment. The leachate was obtained from a landfill in Tetlama, Morelos (Mexico) during the drought season and was characterized in terms of its major pollutants. Considerable levels of chemical oxygen demand (COD), total carbon (TC) and NH4+ were identified, as well as high concentrations of Hg, Pb, and As. Other heavy metals such as Ni, Co, Zn, Cd, and Mn were detected at trace levels. The lethal concentration (LC50) of the leachate, evaluated on Artemia salina, was 12,161±11 mg/L of COD, demonstrating an antagonistic interaction among the leachate's components. The treatment of this effluent consisted of a coagulation-flocculation process using an optimal dose of FeCl3 · 6H2O of 300 mg/L. The supernatant was treated using a photo-Fenton process mediated with FeCl2 · 4H2O and H2O2 in a compound parabolic concentrator (CPC) photo-reactor operating in batch mode using an R ratio (R=[H2O2]/[Fe2+]) of 114. The global removal efficiencies after treatment were 56% for the COD, 95% for TC, and 64% for NH4+. The removal efficiencies for As, Hg, and Pb were 46%, 9%, and 85%, respectively.

NH4+-NH3 removal from simulated wastewater using UV-TiO2 photocatalysis: effect of co-pollutants and pH.

The main objective of the present study was to investigate the efficiency of titanium dioxide (TiO2) assisted photocatalytic degradation (PCD) process for the removal of ammonium-ammonia (NH4(+)-NH3) from the aqueous phase and in the presence of co-pollutants thiosulfate (S2O3(2-)) and p-cresol (C6H4CH3OH) under varying mixed conditions. For the NH4(+)-NH3 only PCD experiments, results showed higher NH4 -NH3 removal at pH 12 compared to pH 7 and 10. For the binary NH4(+)-NH3/S2O3(2-) studies the respective results indicated a significant lowering in NH4(+)-NH3 PCD in the presence of S2O32- at pH 7/12 whereas at pH 10 a marked increase in NH4(+)-NH3 removal transpired. A similar trend was noted for the p-cresol/NH4(+)-NH3 binary system. Comparing findings from the binary (NH4(+)-NH3/S2O3(2-) and p-cresol/NH4(+)-NH3) and tertiary (NH4(+)-NH3/S2O3(2-)/p-cresol) systems, at pH 10, showed fastest NH4(+)-NH3 removal transpiring for the tertiary system as compared to the binary systems, whereas both the binary systems indicated comparable NH4(+)-NH3 removal trends. The respective details have been discussed.

Color change capacity of dental resin mixed with silver methacrylate caused by light irradiation and heating.

We synthesized silver methacrylate, added it to resin for dental use, and observed color changes in the resin from colorless to red-purple following light irradiation. Therefore, 0.01-0.10 wt% silver methacrylate was added to two types of resin powder, i.e., self-curing resin produced by the polymerization of a barbituric acid derivative, quaternary ammonium, and an organometallic compound (Type I), and BPO-tertiary amine resin (Type II), and samples were produced by polymerization with the attached monomer. Changes with time in the color of samples from 3 hours (new samples) to 16 days (old samples) after polymerization and also color changes of old and new samples after heating were evaluated. Subsequently, changes in color with variations in the light intensity, irradiation time, and wavelength were evaluated in terms of color differences.

Conformation and dynamics of DNA molecules during photoreversible condensation.

Direct observation of the mechanism and dynamics of photo-initiated DNA compaction and re-expansion using a light-responsive cationic surfactant has been achieved with fluorescence microscopy. The surfactant undergoes a reversible photoisomerization upon exposure to visible (trans isomer, relatively hydrophobic) or UV (cis isomer, relatively hydrophilic) light. Thus, surfactant binding to DNA and the DNA condensation that result can both be initiated and controlled with light illumination. The inherent kinetics of DNA conformational changes, directly visualized following the in situ light "trigger" of surfactant photoisomerization, are found to occur at rates of approximately 9 microm/s or 240 kbp/s, at or near rates that can be achieved in natural processes. Furthermore, observation of photo-initiated DNA compaction, free of the effects of shear or mixing, provides evidence of a condensation mechanism that nucleates at the ends of the macromolecule. Ethidium bromide displacement studies, employed to gain insight on the mode of interaction between the photo-surfactant and DNA, also reveal the importance of both electrostatic and hydrophobic forces in surfactant binding and DNA condensation.

Assessment of cationic surfactants mineralization by ozonation and photo-Fenton process.

Aqueous solutions of two important quaternary ammonium compounds--16-BAC (benzyl-dimethyl-hexadecylammonium-chloride) and 18-BAC (benzyl-dimethyl-stearylammonium-chloride)--were treated by the ozonation and photo-Fenton processes at different ozone doses and hydrogen peroxide concentrations, respectively. During the photo-Fenton experiments, two different types of lamps were used--a UV mercury vapor medium pressure lamp and a xenon lamp, which simulates solar radiation. The total organic carbon removal was monitored to follow the mineralization of the surfactants. According to the experimental results, after 90 minutes of treatment, the photo-Fenton process achieved up to 80% of mineralization when the UV lamp was used. The efficiency of the photo-Fenton with the xenon lamp was lower. The ozonation process reached, at most, 50% mineralization at the used conditions (ozone dose = 7.57 g/h).

Effect of light on self-assembly of aqueous mixtures of sodium dodecyl sulfate and a cationic, bolaform surfactant containing azobenzene.

We report light and small-angle neutron scattering measurements that characterize microstructures formed in aqueous surfactant solutions (up to 1.0 wt % surfactant) containing mixtures of sodium dodecyl sulfate (SDS) and the light-sensitive bolaform surfactant, bis(trimethylammoniumhexyloxy)azobenzene dibromide (BTHA) as a function of composition, equilibration time, and photostationary state (i.e., solutions rich in cis-BTHA or trans-BTHA). We observed formation of vesicles in both SDS-rich and trans-BTHA-rich regions of the microstructure diagram, with vesicles present over a particularly broad range of compositions for trans-BTHA-rich solutions. Illumination of mixtures of BTHA and SDS with a broadband UV light source leads to formation of photostationary states where the fraction of BTHA present as cis isomer (75-80% cis-BTHA) is largely independent of the mixing ratio of SDS and BTHA. For a relatively limited set of mixing ratios of SDS and BTHA, we observed UV illumination of SDS-rich vesicles to result in the reversible transformation of the vesicles to micellar aggregates and UV illumination of BTHA-rich vesicles to result in irreversible precipitation. Surprisingly, however, for many mixtures of trans-BTHA and SDS that formed solutions containing vesicles, illumination with UV light (which was confirmed to lead to photoisomerization of BTHA) resulted in only a small decrease in the number of vesicles in solution, relatively little change in the sizes of the remaining vesicles, and coexistance of the vesicles with micelles. These observations are consistent with a physical model in which the trans and cis isomers of BTHA present at the photostationary state tend to segregate between the different microstructures coexisting in solution (e.g., vesicles rich in trans-BTHA and SDS coexist with micelles rich in cis-BTHA and SDS). The results presented in this paper provide guidance for the design of light-tunable surfactants systems.

Probing lysozyme conformation with light reveals a new folding intermediate.

A means to control lysozyme conformation with light illumination has been developed using the interaction of the protein with a photoresponsive surfactant. Upon exposure to the appropriate wavelength of light, the azobenzene surfactant undergoes a reversible photoisomerization, with the visible-light (trans) form being more hydrophobic than the UV-light (cis) form. As a result, surfactant binding to the protein and, thus, protein unfolding, can be tuned with light. Small-angle neutron scattering (SANS) measurements were used to provide detailed information of the protein conformation in solution. Shape-reconstruction methods applied to the SANS data indicate that under visible light the protein exhibits a native-like form at low surfactant concentrations, a partially swollen form at intermediate concentrations, and a swollen/unfolded form at higher surfactant concentrations. Furthermore, the SANS data combined with FT-IR spectroscopic analysis of the protein secondary structure reveal that unfolding occurs primarily in the alpha domain of lysozyme, while the beta domain remains relatively intact. Thus, the surfactant-unfolded intermediate of lysozyme appears to be a separate structure than the well-known alpha-domain intermediate of lysozyme that contains a folded alpha domain and unfolded beta domain. Because the interactions between the photosurfactant and protein can be tuned with light, illumination with UV light returns the protein to a native-like conformation. Fluorescence emission data of the nonpolar probe Nile red indicate that hydrophobic domains become available for probe partitioning in surfactant-protein solutions under visible light, while the availability of these hydrophobic domains to the probe decrease under UV light. Dynamic light scattering and UV-vis spectroscopic measurements further confirm the shape-reconstruction findings and reveal three discrete conformations of lysozyme. The results clearly demonstrate that visible light causes a greater degree of lysozyme swelling than UV light, thus allowing for the protein conformation to be controlled with light.

Encoding light intensity by the cone photoreceptor synapse.

How cone synapses encode light intensity determines the precision of information transmission at the first synapse on the visual pathway. Although it is known that cone photoreceptors hyperpolarize to light over 4-5 log units of intensity, the relationship between light intensity and transmitter release at the cone synapse has not been determined. Here, we use two-photon microscopy to visualize release of the synaptic vesicle dye FM1-43 from cone terminals in the intact lizard retina, in response to different stimulus light intensities. We then employ electron microscopy to translate these measurements into vesicle release rates. We find that from darkness to bright light, release decreases from 49 to approximately 2 vesicles per 200 ms; therefore, cones compress their 10,000-fold operating range for phototransduction into a 25-fold range for synaptic vesicle release. Tonic release encodes ten distinguishable intensity levels, skewed to most finely represent bright light, assuming release obeys Poisson statistics.

Photocontrol of protein folding: the interaction of photosensitive surfactants with bovine serum albumin.

The photoresponsive interaction of light-sensitive azobenzene surfactants with bovine serum albumin (BSA) at neutral pH has been investigated as a means to control protein folding with light irradiation. The cationic azobenzene surfactant undergoes a reversible photoisomerization upon exposure to the appropriate wavelength of light, with the visible-light (trans) form of the surfactant being more hydrophobic than the UV-light (cis) form. As a consequence, the trans form exhibits enhanced interaction with the protein compared to the cis form of the surfactant, allowing photoreversible control of the protein folding/unfolding phenomena. Small-angle neutron-scattering (SANS) measurements are used to provide detailed information on the protein conformation in solution. A fitting of the protein shape to a low-resolution triaxial ellipsoid model indicates that three discrete forms of the protein exist in solution depending on the surfactant concentration, with lengths of approximately 90, 150, and 250 A, respectively, consistent with additional dynamic light-scattering measurements. In addition, shape-reconstruction methods are applied to the SANS data to obtain relatively high-resolution conformation information. The results confirm that BSA adopts a heart-shaped structure in solution at low surfactant concentration, similar to the well-known X-ray crystallographic structure. At intermediate surfactant concentrations, protein elongation results as a consequence of the C-terminal portion separating from the rest of the molecule. Further increases in the surfactant concentration eventually lead to a highly elongated protein that nonetheless still exhibits some degree of folding that is consistent with the literature observations of a relatively high helical content in denatured BSA. The results clearly demonstrate that the visible-light form of the surfactant causes a greater degree of protein unfolding than the UV-light form, providing a means to control protein folding with light that, within the resolution of SANS, appears to be completely reversible.

Fast-neutron OSL sensitivity of thallium-doped ammonium salts.

The main problem in selecting suitable thermoluminescent (TL) materials for fast-neutron dosimetry is finding a material that is both tissue-equivalent and not damaged upon heating. Optically stimulated luminescence (OSL) avoids the need to heat the materials and allows the use of materials with a high content of hydrogen (responsible for 90% of the absorbed dose of fast-neutrons). The choice of studying the ammonium salts for their OSL properties was based on the calculation of their neutron kerma factor. A constant ratio of an ammonium salt's kerma coefficients to the tissue's kerma coefficients (in the fast-neutron range) is a prerequisite for a similar energy response to neutrons, i.e. tissue equivalency. The salts studied are NH4Br and (NH4)2SiF6 both doped with Tl+. This paper describes the OSL properties of Tl(+)-doped NH4Br and (NH4)2SiF6 after exposure to 14.5 MeV neutrons to explore their potential for developing new, tissue-equivalent OSL materials suitable for fast-neutron dosimetry. The relative neutron sensitivity, k, defined as the ratio of the sensitivity of the material to neutrons to its sensitivity to gamma rays, has been determined for 14.5 MeV neutrons and varies between k = 0.15 and k = 0.5. The latter value is a factor 2.5 higher than that found for known TL materials (k < or = 0.2). A drawback of these materials is the fast fading of the OSL signal.