Probe Diffusion Research Articles

Polymer Conformations and Diffusion through a Monolayer of Confining Nanoparticles

We present coarse-grained molecular dynamics simulations to probe chain-scale polymer conformations and diffusion between confining nanoparticles (NPs). By constructing a monolayer of hexagonally packed NPs in a polymer melt with athermal interactions, we observe the magnitude and length scale over which homogeneously confining NPs impact the polymer behavior, which provides fundamental insights into more complex polymer nanocomposites. We show that polymer conformations are more perturbed under strong confinement (i.e., when the interparticle distance, ID, is less than twice the polymer radius of gyration, 2Rg) as compared to around an isolated NP, and the effect depends on the ratio of RNP/Rg rather than either independently. In fact, these conformations can be quantitatively replicated by executing a simple random walk in a similarly confining environment. We also show that polymer diffusion is slowed by the presence of NPs, and the slowing persists far beyond the length scale over which polymer conformations are perturbed. Although the slowing is strongest ∼Rg from the NPs, the diffusion coefficient is slower even beyond ∼5Rg from the NPs. Furthermore, by analyzing the directional van Hove distributions, we show polymer diffusion away from the NP monolayer is bulk-like while diffusion through the monolayer is slower with increasing NP confinement. These molecular dynamics simulations provide fundamental insights into the temporal and spatial effect of confining, athermal NPs on chain-scale polymer conformations, and diffusion.

Miniaturized saturated absorption spectrometer

We describe a saturated absorption spectrometer that is robust and compact and requires minimum alignment, which is made possible by using a diffuse probe beam generated by a retro-reflecting film. This concept was studied and implemented in a miniaturized home-built setup that provides the same performance as an optimized table-top setup.

A comprehensive model for the diffusion and hybridization processes of nucleic acid probes in fluorescence in situ hybridization.

Fluorescence in situ hybridization (FISH) has been extensively used in the past decades for the detection and localization of microorganisms. However, a mechanistic approach of the whole FISH process is still missing, and the main limiting steps for the hybridization to occur remain unclear. In here, FISH is approached as a particular case of a diffusion-reaction kinetics, where molecular probes (MPs) move from the hybridization solution to the target RNA site within the cells. Based on literature models, the characteristic times taken by different MPs to diffuse across multiple cellular barriers, as well as the reaction time associated with the formation of the duplex molecular probe-RNA, were estimated. Structural and size differences at the membrane level of bacterial and animal cells were considered. For bacterial cells, the limiting step for diffusion is likely to be the peptidoglycan layer (characteristic time of 7.94 × 102 - 4.39 × 103 s), whereas for animal cells, the limiting step should be the diffusion of the probe through the bulk (1.8-5.0 s) followed by the diffusion through the lipid membrane (1 s). The information provided here may serve as a basis for a more rational development of FISH protocols in the future.

Muon Spectroscopy for Investigating Diffusion in Energy Storage Materials

We review recent applications of positive muon spin relaxation (μSR) spectroscopy as an active probe of ion diffusion in energy storage materials. μSR spectroscopy allows the study of ionic diffusion in solid-state materials on a time scale between 10−5 and 10−8 s where most long-range and consecutive short-range jumps of ions between interstitial sites occur. μSR also allows one to probe and model ionic diffusion in materials that contain magnetic ions, since both electronic and nuclear contributions to the muon depolarization can be separated, making μSR an excellent technique for the microscopic study of the ionic motions in crystalline materials. We highlight a series of battery materials for which μSR has provided insight into intrinsic ionic conduction and magnetic properties without interference of external factors, such as the presence of magnetic ions, macroscopic particle morphologies, or elaborate measurement setups.

Nanoparticle diffusion in polymer melts: Molecular dynamics simulations and mode-coupling theory.

Nanoparticle diffusion in polymer melts is studied by the combination of Molecular Dynamics (MD) simulations and Mode-Coupling Theory (MCT). In accord with earlier experimental, simulation, and theoretical studies, we find that the Stokes-Einstein (SE) hydrodynamic relation Dn ∼ 1/Rn holds when the nanoparticle radius Rn is greater than the polymer gyration radius Rg, while in the opposite regime, the measured nanoparticle diffusion coefficient Dn exceeds the SE value by as much as an order of magnitude. The MCT values of Dn are found to be consistently higher than the MD simulation values. The observed discrepancy is attributed to the approximations involved in constructing the microscopic friction as well as to the approximate forms for dynamic structure factors used in MCT. In a thorough test of underlying MCT assumptions and approximations, various structural and dynamical quantities required as input for MCT are obtained directly from MD simulations. We present the improved MCT approach, which involves splitting of the microscopic time-dependent friction into two terms: binary (originating from short-time dynamics) and collective (due to long-time dynamics). Using MD data as input in MCT, we demonstrate that the total friction is largely dominated by its binary short-time term, which, if neglected, leads to severe overestimation of Dn. As a result, the revised version of MCT, in agreement with the present MD data, predicts 1/Rn 2 scaling of the probe diffusion coefficient in a non-hydrodynamic regime when Rn < Rg. If the total friction is dominated by the collective long-time component, one would observe 1/Rn 3 scaling of Dn in accordance with previous studies.

Permeability of Dawson–type polyoxometalates through vertically oriented nanoporous silica membranes on electrode: Effect of pore size and probe charge

The voltammetric behavior of bulky heteropolyanions belonging to the Dawson family of polyoxometalates, [P2VmW18-mO62]n− (m = 0, 3, 6), has been investigated at electrodes coated with vertically-aligned mesoporous silica thin films with pore openings in the 2–3 nm range. The experiments have been carried out in acidic medium (pH 0.3–1), where the silica surface is positively charged and thus likely to interact with the negatively charged redox probes, and at various ionic strengths of the medium. Better permeability of the films was clearly observed at lower pH values, corresponding to higher densities of positive charges onto the silica walls, and of films with a larger mesopore size, favoring the mass transport rates through the nanoporous membrane. Diffusion of such bulky anionic redox probes was dramatically influenced by the ionic strength of the medium, being especially restricted at low electrolyte concentrations where the thickness of the electrical double layer became of the same order of magnitude or larger than the mesopore radius, giving rise to very low voltammetric signals. The overall negative charge on POMs had few effects on their mass transport through the film, except at low ionic strength where the most highly charged redox probes gave rise to the lowest current responses.

Optimizing the Atomic Layer Deposition of Alumina on Perovskite Nanocrystal Films by Using O2 As a Molecular Probe

AbstractEncapsulation methods have shown to be effective in imparting improved stability to metal‐halide perovskite nanocrystals (NCs). Atomic layer deposition (ALD) of metal oxides is one of the promising approaches for such encapsulation, yet better control on the process parameters are required to achieve viable lifetimes for several optoelectronic and photocatalytic applications. Herein, we optimize the ALD process of amorphous aluminum oxide (AlOx) as an encapsulating layer for CsPbBr3 NC thin films by using oxygen (O2) as a molecular diffusion probe to assess the uniformity of the deposited AlOx layer. When O2 reaches the NC surface, it extracts the photogenerated electrons, thus quenching the PL of the CsPbBr3 NCs. As the quality of the ALD layer improves, less quenching is expected. We compare three different ALD deposition modes. We find that the low temperature/high temperature and the exposure modes improve the quality of the alumina as a gas barrier when compared with the low temperature mode. We attribute this result to a better diffusion of the ALD precursor throughout the NC film. We propose the low temperature/high temperature as the most suitable mode for future implementation of multilayered coatings.

Adaptability of Granier empirical formula in sap flow measurement of Populus tomentosa based on whole tree weighing method

We evaluated the adaptability of Granier's empirical formula in the measurement of trunk sap flow in Populus tomentosa. The thermal diffusion probe method (TDP) was used to mea-sure sap flow rate, and the whole tree weighing was simultaneously measured for each tree. We compared results from the Granier empirical formula with that from the whole tree weighing to find out whether Granier formula had any error in measuring the trunk sap flow of P. tomentosa. The transpiration rate by the whole tree weighing method and the temperature difference coefficient K by the thermal diffusion method were fitted with power exponential regression to establish a corrected Granier formula. Compared with the transpiration rate measured by the whole tree weighing method, sap flow rate calculated by the Granier empirical formula was underestimated by 67.7%. Therefore, a calibrated Granier correction formula of P. tomentosa was established: Fd=0.0135K0.6952(R2=0.77). The calculated result from this calibrated formula was only 3.4% lower than the transpiration rate estimated with the whole tree weighing method, which showed good consistency. Thus, the calculation of the P. tomentosa sap flow rate should be corrected when using the Granier empirical formula.

Electron Paramagnetic Resonance Measurements of Four Nitroxide Probes in Supercooled Water Explained by Molecular Dynamics Simulations.

Electron paramagnetic resonance (EPR) measurements of the rotational diffusion of small nitroxide probes have been demonstrated to be a powerful technique for experimentally investigating the properties of supercooled liquids, such as water. However, since only the rotational diffusion of the probe molecules is measured and EPR measurements are indirect, it is not clear what the relationship between the behavior of water and the probe molecule is. To address this, we have performed molecular dynamics simulations of four nitroxide probes in TIP4P-Ew and OPC water models to directly compare with EPR experiments and to determine the behavior of the water and the underlying microscopic coupling between the water and the probes. In all, 200 ns simulations were run for 23 temperatures between 253 and 283 K for all four probes with each water model for an aggregate of 36.8 μs of simulation time. Simulations for both water models systematically underestimated the rotational diffusion coefficients for both water and probes, though OPC simulations were generally in better agreement with the experiments than TIP4P-Ew simulations. Despite this, when the temperature dependence of the data was fit to a power law, fit parameters for TIP4P-Ew were generally in better agreement with the experiments than OPC. For probe molecules, the singular temperature was found to be T0 = 226.5 ± 0.4 K from experiments, T0 = 208 ± 2 K for OPC water, and T0 = 215 ± 2 K for TIP4P-Ew water. While for water molecules, the singular temperature was found to be T0 = 220.3 ± 0.2 K from experiments, T0 = 208 ± 2 K for OPC water, and T0 = 220 ± 1 K for TIP4P-Ew water. Systematic underestimation of the rotational diffusion coefficients was most pronounced at lower temperatures and was clearly observed in changes to the Arrhenius activation energy. Above the maximum density temperature of Tρmax = 277 K, an activation energy of EA ≈ 16.7 kJ/mol was observed for the probes from experiments, while OPC had EA ≈ 15.2 kJ/mol and TIP4P-Ew had EA ≈ 14.6 kJ/mol. Below the maximum density temperature, the activation energy jumped to EA ≈ 32.5 kJ/mol for experiments but only EA ≈ 23 kJ/mol for OPC and EA ≈ 22 kJ/mol for TIP4P-Ew. In all cases, we saw good agreement between the behavior of the probe molecules and water. To understand why, we calculated the average number of hydrogen bonds between the probe molecules and water. From this, we were able to explain the rotational diffusion times for all of the probes. These results show that current molecular models are sufficient to capture physical phenomena observed with EPR and to help elucidate why the probes provide accurate insights into the behavior of supercooled water.

Agarose Hydrogels Enriched by Humic Acids as the Complexation Agent.

The transport properties of agarose hydrogels enriched by humic acids were studied. Methylene blue, rhodamine 6G and Cu(II) ions were incorporated into hydrogel as diffusion probes, and then their release into water was monitored. Cu(II) ions as well as both the dyes studied in this work have high affinity to humic substances and their interactions strongly affected their diffusion in hydrogels. It was confirmed that humic acids retarded the transport of diffusion probes. Humic acids’ enrichment caused the decrease in the values of effective diffusion coefficients due to their complexation with diffusion probes. In general, the diffusion of dyes was more affected by the complexation with humic acids in comparison with Cu(II) ions. The effect of complexation was selective for the particular diffusion probe. The strongest effect was obtained for the diffusion of methylene blue. It was assumed that metal ions interacted preferentially with acidic functional groups. In contrast to Cu(II) ions, dyes can interact with acidic functional groups, and the condensed cyclic structures of the dye probes supported their interactions with the hydrophobic domains of humic substances.

Probing the microenvironment of polyacrylamide hydrogel matrix using turbidity and fluorescence recovery after photobleaching: One versus Two phases

The microstructure of polyacrylamide hydrogel matrices was probed by turbidity measurements, and by fluorescence recovery after photobleaching (FRAP) of fluorescently labeled bovine serum albumin (fBSA), as a function of polymer concentration and crosslink density. Turbidity increased with increasing polymer and/or crosslinker concentration. At low polymer and crosslinker concentrations, FRAP dynamics were well described by diffusion through a single polymer phase. However for higher concentrations of polymer and crosslinker, the diffusion model was inadequate, and a second mode, likely corresponding to release of fBSA from regions of relatively dense polymer, was observed. At low network concentrations, probe diffusivity (inversely related to FRAP relaxation time) decreased with increasing polymer and crosslinker concentrations. Following onset of the second mode, however, diffusivity increased with increasing monomer concentration, suggesting that the network through which fBSA diffused became looser. The optical and FRAP measurements, while showing qualitatively similar trends, were not directly correlated, and they likely reflected different modes of network phase separation.

Density of Obstacles Affects Diffusion in Adsorbed Polymer Layers.

The translational diffusion of molecules dispersed into polymer matrices slows down tremendously when approaching a nonrepulsive interface. To unravel the origin of this phenomenon, we investigated the diffusion of molecular probes in the direction normal to an adsorbing wall. Using adsorbed polymer layers as matrices, wewere able todecouple interfacial and finite size effects and determined the relation between the diffusion time and the area available at the polymer/solid interface. Based on the results of our investigation, we present a physical picture, suggesting that the reduction indiffusion rate is correlated to the degree of chain adsorption onto the substrate, that is, the density of surface obstacles encountered by tracer molecules.

Interstitial diffuse optical probe with spectral fitting to measure dynamic tumor hypoxia

Understanding the dynamic nature of tumor hypoxia is vital for cancer therapy. The presence of oxygen within a tumor during radiation therapy increases the likelihood of local control. We used a novel interstitial diffuse optical probe to make real-time measurements of blood volume fraction and hemoglobin oxygen saturation within a tumor at a high temporal resolution. This device was initially characterized and benchmarked using a customized vessel designed to control hemoglobin oxygen saturation and blood volume in a solution of blood with different concentrations of an oxygen scavenger, tetrakis (hydroxymethyl) phosphonium chloride. The optical device was found to consistently monitor the changes in oxygen saturation and these changes correlated to the concentration of the oxygen scavenger added. In near-simultaneous measurements of blood volume and oxygen saturation in tumor-bearing mice, the changes in blood volume fraction and oxygen saturation measured with the interstitial diffuse optical probe were benchmarked against photoacoustic imaging system to track and compare temporal dynamics of oxygen saturation and blood volume in a patient-derived xenograft model of hypopharyngeal carcinoma. Positive correlations between our device and photoacoustic imaging in measuring blood volume and oxygen saturation were observed.

An intramolecular catalytic hairpin assembly on a DNA tetrahedron for mRNA imaging in living cells: improving reaction kinetics and signal stability.

Enzyme-free amplification techniques based on dynamic DNA self-assembly (DDSA) have recently been developed for the in situ detection of mRNA in living cells. However, signal generation in traditional DDSA amplifiers is mainly dependent on the random diffusion of dissociative probes in a bulk solution, which is generally accompanied by poor kinetics and interference from complex biological systems. In this work, a new amplifier based on the design of an intramolecular catalytic hairpin assembly (intra-CHA) is proposed for the FRET imaging of mRNA in living cells. Compared with that in the free catalytic hairpin assembly (free-CHA), probes H1 and H2 in intra-CHA were simultaneously fixed on a DNA tetrahedron. The distance between them was closer, the local concentration of H1 and H2 in intra-CHA was theoretically approximately 808-times higher than that in free-CHA, and the initial reaction rate was enhanced 15.6 fold. Due to the spatial confinement effect, the reaction kinetics for target-catalyzed signal generation were significantly improved. By virtue of the three-dimensional nanostructure, H1 and H2 in the intra-CHA amplifier entered cells without any transfection or nanocarrier, and the probes and their products were free from biological interference, providing much higher signal stability for the reliable imaging of mRNA in living cells.

Translation of a portable diffuse optical breast scanner probe for clinical application: a preliminary study

Most breast cancer lesions absorb higher levels of near-infrared (NIR) radiation compared to healthy breast tissue due to its increased vascularity. Oxy-hemoglobin (HbO2) and deoxy-hemoglobin (Hb) primarily found in cancerous vascular lesions, absorbs higher levels of radiation in the 650 nm to 850 nm wavelength range than the surrounding fatty tissue and water in the human breast. NIR diffuse optical spectroscopy (DOS) provides real-time functional and compositional information based on the optical properties of biological tissues, which cannot be accomplished by other portable breast imaging modalities. Here we present the first set of clinical trials using a non-invasive, hand-held diffuse optical breast scanner (DOB-Scan probe) to capture in vivo cross-sectional images of the breast. The scanner uses four NIR illuminating sources with different wavelengths, 690 nm, 750 nm, 800 nm, and 850 nm, to determine the concentrations of the four main constituents of breast tissue, oxy-hemoglobin (HbO2), deoxy-hemoglobin (Hb), water (H2O), and fat. In this paper, we briefly explain the hardware design and image reconstruction algorithm of the DOB-Scan probe, the data collection process, and the imaging results of four different participants, selected from twenty, all who are diagnosed with breast cancer. For each patient, images were scanned from two locations, the first over the cancerous lesion and the second over the same region on the contralateral healthy breast, as a means of establishing controls for comparison. During each scan, four cross-sectional images of the breast, corresponding to four different NIR wavelengths, are reconstructed and displayed on a user interface for reference. Clinical results confirm that the absorption coefficients of cancerous lesions are significantly higher than the normal surrounding tissue. We propose to deploy the probe to effectively identify cancerous breast tissue at an early stage in a primary care setting, which could increase the efficiency of screening programs.

Diffusion and flow in complex liquids.

Thermal motion of particles and molecules in liquids underlies many chemical and biological processes. Liquids, especially in biology, are complex due to structure at multiple relevant length scales. While diffusion in homogeneous simple liquids is well understood through the Stokes-Einstein relation, this equation fails completely in describing diffusion in complex media. Modeling, understanding, engineering and controlling processes at the nanoscale, most importantly inside living cells, requires a theoretical framework for the description of viscous response to allow predictions of diffusion rates in complex fluids. Here we use a general framework with the viscosity η(k) described by a function of wave vector in reciprocal space. We introduce a formulation that allows one to relate the rotational and translational diffusion coefficients and determine the viscosity η(k) directly from experiments. We apply our theory to provide a database for rotational diffusion coefficients of proteins/protein complexes in the bacterium E. coli. We also provide a database for the diffusion coefficient of proteins sliding along major grooves of DNA in E. coli. These parameters allow predictions of rate constants for association of proteins. In addition to constituting a theoretical framework for description of diffusion of probes and viscosity in complex fluids, the formulation that we propose should decrease substantially the cost of numerical simulations of transport in complex media by replacing the simulation of individual crowding particles with a continuous medium characterized by a wave-length dependent viscosity η(k).

模拟穿透雨减少对锐齿栎(Quercus aliena var. acuteserrata)树干液流密度的影响

PDF HTML阅读 XML下载 导出引用 引用提醒 模拟穿透雨减少对锐齿栎(Quercus aliena var. acuteserrata)树干液流密度的影响 DOI: 10.5846/stxb201903060423 作者: 作者单位: 作者简介: 通讯作者: 中图分类号: 基金项目: 国际竹藤中心基本科研业务费专项基金项目（1632018002，1632018004，1632017016）；国家重点研发计划（2018YFC0507301） Effects of manipulated throughfall reduction on sap flux density of Quercus aliena var. acuteserrata Author: Affiliation: Fund Project: International Bamboo and Rattan Center"s special funds for basic research and operation fees;National Key R&D Program 摘要 | 图/表 | 访问统计 | 参考文献 | 相似文献 | 引证文献 | 资源附件 | 文章评论 摘要:降水格局变化是全球气候变化的重要特征之一，未来气候变化下，较为频繁和严峻的干旱将威胁地球中纬度部分地区的森林，但森林植被如何响应季节性干旱胁迫及其机制尚不清楚。北亚热带-暖温带过渡区分布着以锐齿栎（Quercus aliena var. acuteserrata）为优势树种的落叶阔叶林，研究其水分蒸腾代谢过程对干旱的响应是评估气候变化对过渡区天然落叶阔叶林生态系统水碳影响的关键科学问题。在典型的锐齿栎天然林中通过开展模拟穿透雨减少大型野外实验，采用Granier热扩散式探针技术监测锐齿栎树干液流密度的动态变化，研究了不同径级锐齿栎树干液流密度对模拟干旱的响应规律。结果表明：（1）穿透雨减少对树干液流密度的影响呈现季节变异。在7月份，林内穿透雨减少显著降低了锐齿栎的树干液流密度，但生长季后期的10月份林内穿透雨减少反而使锐齿栎树干液流密度显著升高。（2）不同径级的锐齿栎树干液流密度在生长季内对干旱有不同的响应，特别是小径级的树干液流密度与其他径级有较多的不同。小径级的锐齿栎树干液流密度在5、7月份表现为减雨样地显著小于对照样地，在9、10月份则表现为减雨样地显著大于对照样地。中径级的锐齿栎树干液流密度在5、10月份表现为减雨样地显著大于对照样地，在7月份则表现为减雨样地极显著小于对照样地。大径级的锐齿栎树干液流密度在6、7月份表现为减雨样地显著小于对照样地，在10月份则表现为减雨样地显著大于对照样地。 Abstract:The change in precipitation pattern is one of the characteristics of global climate change. Some regions at mid-latitude might experience more frequent and severe drought. However, it is unclear how the precipitation change will affect the forest vegetation in those regions. Amongst, Quercus aliena var. acuteserrata is the dominant canopy tree species of deciduous broad-leaved forests in the warm temperate zone of middle China. Understanding the evapotranspiration responses of Q. aliena to drought induced by climate change is crucial to elucidate carbon-water coupling process. In order to explore the warm-temperate deciduous broad-leaved forest response to precipitation change, a manipulated throughfall reduction experiment was conducted in Baotianman National Nature Reserve since 2013. The Granier thermal diffusion probe (TDP) was used to measure the dynamic change of sap flux density of Q. aliena. The results showed that the throughfall reduction had significant effects on the sap flux density (Js) during the growing season. Firstly, the throughfall reduction significantly reduced the sap flux density of Q. aliena in July. However, in the late growing season (October), the throughfall reduction increased the sap flux density of Q. aliena significantly. Therefore, the sap flux density of Q. aliena differently responded to the throughfall reduction among months during the growing season. Secondly, the sap flux density of Q. aliena of different tree sizes showed different responses to drought during the growing season, especially the small trees (diameter at breast height, DBH≤21 cm) was different from that of medium (21 cm 参考文献 相似文献 引证文献

Multimodal Nanocarrier Probes Reveal Superior Biodistribution Quantification by Isotopic Analysis over Fluorescence.

Absolute measurements of biodistribution are essential for understanding and optimizing the function of nanomaterials for in vivo diagnostic and therapeutic applications. Biodistribution analysis by optical imaging is desirable due to its low cost, wide accessibility, and high-throughput nature, but it is substantially less accurate than isotopic and chemical techniques. In this work, we developed multimodal probes for optical and nuclear imaging to analyze the quantitative limits of optical contrast in the red and near-infrared spectra for polysaccharide nanocarriers targeting macrophage cells. Probes incorporating three zwitterionic fluorophores together with radioactive copper distributed diffusely to optically dissimilar tissues that were either white (visceral adipose tissue) or dark red (liver and spleen) in obese rodents. We used in vivo positron emission tomography/computed tomography (PET/CT) imaging, in vivo hyperspectral tomographic fluorescence imaging, and ex vivo optical and isotopic analyses to determine correlations between optical and nuclear signals. PET imaging strongly correlated with standardized ex vivo methods for all tissue types, whereas no fluorescence signals exhibited substantial accuracy in quantification or localization in vivo. Optical imaging of resected tissues was most accurate in the 700 nm wavelength window, but only in white tissues. This work suggests that fluorescence can be used to measure diffuse probe distribution in white tissues over time or across animals, but not red tissues and not deep in the body. This work also highlights the importance of choosing validated experimental protocols and describes how optical measurements are impacted by fluorophore class and spectral properties, tissue properties, and imaging workflow.

Artificial membranes in combination with selected natural oils for in vitro drug passive diffusion screening in Ussing type chamber apparatus applied to gastro-retentive systems

This study aimed at developing an effective in vitro technique for the screening of drug passive diffusion utilising artificial membranes in combination with three selected oils (i.e. cognac, emu, and olive oil). Artificial membranes of varying chemical composition and characteristics have been investigated individually and in combination with the selected oils in terms of the passive diffusion of a fluorescent probe (i.e. Rhodamine 6G or R6G), in a diffusion apparatus as compared to excised pig intestinal tissues. In general, the permeation results showed that the rate and extent of R6G permeation were dependent on the membrane composition as well as the type of oil used. The apparent permeability coefficient (Papp) value for R6G across the cellulose nitrate membrane (0.197 × 10−7 ± 0.069 cm/s) was the closest to the Papp of R6G across the excised pig intestinal tissue (0.210 × 10−7 ± 0.080 cm/s). The cellulose acetate-nitrate mixture membrane impregnated with emu oil also produced a Papp value (0.191 × 10−7 ± 0.010 cm/s) that was relatively close to that of R6G across the excised pig intestinal tissue. The delivery of R6G from gastro-retentive matrix type tablets correlated with the release of R6G from the gastro-retentive tablets.

Chemical tuning of photoswitchable azobenzenes: a photopharmacological case study using nicotinic transmission.

We have developed photochromic probes for the nicotinic acetylcholine receptor that exploit the unique chemical properties of the tetrafluoroazobenzene (4FAB) scaffold. Ultraviolet light switching and rapid thermal relaxation of the metastable cis configuration are the main drawbacks associated with standard AB-based switches. We designed our photoprobes to take advantage of the excellent thermodynamic stability of the cis-4FAB configuration (thermal half-life > 12 days at 37 °C in physiological buffer) and cis–trans photostationary states above 84%. Furthermore, the well-separated n–π* absorption bands of trans- and cis-4FAB allow facile photoswitching with visible light in two optical channels. A convergent 11-step synthetic approach allowed the installation of a trimethylammonium (TA) head onto the 4FAB scaffold, by means of an alkyl spacer, to afford a free diffusible 4FABTA probe. TAs are known to agonize nicotinic receptors, so 4FABTA was tested on mouse brain slices and enabled reversible receptor activation with cycles of violet and green light. Due to the very long-lived metastable cis configuration, 4FAB in vivo use could be of great promise for long term biological studies. Further chemical functionalization of this 4FAB probe with a maleimide functionality allowed clean cross-linking with glutathione. However, attempts to conjugate with a cysteine on a genetically modified nicotinic acetylcholine receptor did not afford the expected light-responsive channel. Our data indicate that the 4FAB photoswitch can be derivatized bifunctionally for genetically-targeted photopharmacology whilst preserving all the favorable photophysical properties of the parent 4FAB scaffold, however, the tetrafluoro motif can significantly perturb pharmacophore–protein interactions. In contrast, we found that the freely diffusible 4FABTA probe could be pre-set with green light into an OFF state that was biologically inert, irradiation with violet light effectively "uncaged" agonist activity, but in a photoreversible manner. Since the neurotransmitter acetylcholine has fully saturated heteroatom valences, our photoswitchable 4FABTA probe could be useful for physiological studies of this neurotransmitter.