Rotational Correlation Time Research Articles

Relationship between Translational and Rotational Dynamics of Alkyltriethylammonium-Based Ionic Liquids.

1H spin-lattice relaxation experiments have been performed for a series of ionic liquids including bis(trifluoromethanesulfonyl)imide anion and cations of a varying alkyl chain length: triethylhexylammonium, triethyloctylammonium, decyltriethylammonium, dodecyltriethylammonium, triethyltetradecylammonium, and hexadecyltriethylammonium. The relaxation studies were carried out in abroad frequency range covering three orders of magnitude, from 10 kHz to 10 MHz, versus temperature. On the basis of a thorough, quantitative analysis of this reach data set, parameters characterizing the relative, cation-cation, translation diffusion (relative diffusion coefficients and translational correlation times), and rotational motion of the cation (rotational correlation times) were determined. Relationships between these quantities and their dependence on the alkyl chain length were discussed in comparison to analogous properties of molecular liquids. It was shown, among other findings, that the ratio between the translational and rotational correlation times is smaller than for molecular liquids and considerably dependent on temperature. Moreover, a comparison of relative and self-diffusion coefficients indicate correlated translational dynamics of the cations.

Relationship between macroscopic properties of honey and molecular dynamics – temperature effects

1H spin-lattice relaxation studies of different kinds of honey have been performed in a broad frequency range, versus temperature. On the basis of the extensive data set, translation diffusion coefficients and rotational correlation times of water molecules confined in the sugar matrix have been determined. It has turned out that the translation dynamics is of three-dimensional (isotropic) character and its by three orders of magnitude slower compared to water in bulk. This scenario established on the molecular level has been compared with the picture of translation diffusion obtained from viscosity measurements. Systematic discrepancies have been observed for all kinds of honey and their origin has been explained. Activation energies of the translational and rotational motion and ratios between the translational and rotational correlational times have been determined and discussed in the context of the properties of supercooled liquids.

High-Pressure ESR Spectroscopy: On the Rotational Motion of Spin Probes in Pressurized Ionic Liquids.

We report high-pressure (up to 50 MPa) ESR-spectroscopic investigations on the rotational correlation times of the nitroxide radicals 2,2,6,6-tetramethylpiperidine 1-oxyl (TEMPO), 4-hydroxy-2,2,6,6-tetramethylpiperidine 1-oxyl (TEMPOL), and 4-amino-2,2,6,6-tetramethylpiperidine 1-oxyl (ATEMPO) in the ionic liquids 1-ethyl-3-methylimidazolium tetrafluoroborate (emimBF4), 1-butyl-3-methylimidazolium hexafluorophosphate (bmimPF6), 1-butyl-3-methylimidazolium tetrafluoroborate (bmimBF4), 1-methyl-3-octylimidazolium tetrafluoroborate (omimBF4), and 1-methyl-3-octylimidazolium hexafluorophosphate (omimPF6). The activation volumes (38.5–56.6 Å3) determined from pressure dependent rotational diffusion coefficients agree well with the pressure dependent viscosities of the ionic liquids. Experimentally, the fractional exponent of the generalized Stokes–Einstein–Debye relation is found to be close to one.

Confinement dependence of protein-associated solvent dynamics around different classes of proteins, from the EPR spin probe perspective.

Protein function is modulated by coupled solvent fluctuations, subject to the degree of confinement from the surroundings. To identify universal features of the external confinement effect, the temperature dependence of the dynamics of protein-associated solvent over 200-265 K for proteins representative of different classes and sizes is characterized by using the rotational correlation time (detection bandwidth, 10-10-10-7 s) of the electron paramagnetic resonance (EPR, X-band) spin probe, TEMPOL, which is restricted to regions vicinal to protein in frozen aqueous solution. Weak (protein surrounded by aqueous-dimethylsulfoxide cryosolvent mesodomain) and strong (no added crysolvent) conditions of ice boundary confinement are imposed. The panel of soluble proteins represents large and small oligomeric (ethanolamine ammonia-lyase, 488 kDa; streptavidin, 52.8 kDa) and monomeric (myoglobin, 16.7 kDa) globular proteins, an intrinsically disordered protein (IDP, β-casein, 24.0 kDa), an unstructured peptide (protamine, 4.38 kDa) and a small peptide with partial backbone order (amyloid-β residues 1-16, 1.96 kDa). Expanded and condensate structures of β-casein and protamine are resolved by the spin probe under weak and strong confinement, respectively. At each confinement condition, the soluble globular proteins display common T-dependences of rotational correlation times and normalized weights, for two mobility components, protein-associated domain, PAD, and surrounding mesodomain. Strong confinement induces a detectable PAD component and emulation of globular protein T-dependence by the amyloid-β peptide. Confinement uniformly impacts soluble globular protein PAD dynamics, and is therefore a generic control parameter for modulation of soluble globular protein function.

Interaction of surfactants with poloxamers 338 and its effect on some properties of cream base

The aim. Study of the interaction of surfactants with poloxamer 338 (P338) and the effect of P338 on the properties of cream bases.
 Materials and methods. Solutions of the surfactants and P338 as well as cream bases were under study. The average hydrodynamic diameter (Dh) and zeta potential (ζ‑potential) were determined by the light scattering intensity and electrophoretic mobility of micelles. The electron paramagnetic resonance (EPR) spectra of spin probes in micelles, solvents and bases were obtained; the type of spectrum, isotropic constant (AN), rotational correlation times (τ) and anisotropy parameter (ε) were determined. Liquids and cream bases were studied by capillary and rotational viscometry; the flow behaviour and yield stress (t0), dynamic and apparent viscosity (η) as well as the hysteresis (thixotropic) area (AH) were determined. The microstructure of the bases was examined by optical microscopy. The strength of adhesion (Sm) was assessed by the pull-off test, and the absorption of water was studied by dialysis.
 Results. Under the impact of P338 the hydrodynamic diameters of micelles formed by cationic, anionic and nonionic surfactants decreased as well as the absolute values of their ζ‑potential became lower, but the microviscosity of the micelle nuclei increased. There was also a change in the structure of the aggregates of surfactant with fatty alcohols; EPR spectra, which were superpositions characteristic for the lateral phase separation, converted into triplets that indicated the uniform distribution of lipophilic probes in the surfactant phase. When the content of P338 increased to 17 %, the rheological parameters of the bases increased drastically, the flow behaviour and the microstructure changed. The bases had the consistency of cream within temperature range from 25 °C to 70 °C and completely restored their apparent viscosity, which had decreased under shear stress. P338 enhances the adhesive properties of the bases. Due to their microstructure, cream bases have a lower ability to absorb water compared to a solution and gel containing 17 % and 20 % P338, respectively.
 Conclusions. The structure of surfactant micelles and aggregates of surfactants with fatty alcohols changed under impact of P338 due to the interaction of surfactants with P338. As a result of this interaction, at a sufficiently high concentration of P338, the microstructure and flow behaviour of bases changed, their rheological parameters, which remain high at temperatures from 25 °C to 70 °C, increased significantly, and water absorption parameters decreased. The bases with P338 were more adhesive

Nanosized T1 MRI Contrast Agent Based on a Polyamidoamine as Multidentate Gd Ligand.

A linear polyamidoamine (PAA) named BAC-EDDS, containing metal chelating repeat units composed of two tert-amines and four carboxylic groups, has been prepared by the aza-Michael polyaddition of ethylendiaminodisuccinic (EDDS) with 2,2-bis(acrylamido)acetic acid (BAC). It was characterized by size exclusion chromatography (SEC), FTIR, UV–Vis and NMR spectroscopies. The pKa values of the ionizable groups of the repeat unit were estimated by potentiometric titration, using a purposely synthesized molecular ligand (Agly-EDDS) mimicking the structure of the BAC-EDDS repeat unit. Dynamic light scattering (DLS) and ζ-potential analyses revealed the propensity of BAC-EDDS to form stable nanoaggregates with a diameter of approximately 150 nm at pH 5 and a net negative charge at physiological pH, in line with an isoelectric point <2. BAC-EDDS stably chelated Gd (III) ions with a molar ratio of 0.5:1 Gd (III)/repeat unit. The stability constant of the molecular model Gd-Agly-EDDS (log K = 17.43) was determined as well, by simulating the potentiometric titration through the use of Hyperquad software. In order to comprehend the efficiency of Gd-BAC-EDDS in contrasting magnetic resonance images, the nuclear longitudinal (r1) and transverse (r2) relaxivities as a function of the externally applied static magnetic field were investigated and compared to the ones of commercial contrast agents. Furthermore, a model derived from the Solomon–Bloembergen–Morgan theory for the field dependence of the NMR relaxivity curves was applied and allowed us to evaluate the rotational correlation time of the complex (τ = 0.66 ns). This relatively high value is due to the dimensions of Gd-BAC-EDDS, and the associated rotational motion causes a peak in the longitudinal relaxivity at ca. 75 MHz, which is close to the frequencies used in clinics. The good performances of Gd-BAC-EDDS as a contrast agent were also confirmed through in vitro magnetic resonance imaging experiments with a 0.2 T magnetic field.

Stimuli-sensitivity and dynamics in the self-assembly structure of TEMPO-containing nonamphiphilic nanoparticles and their triggering hydrophobic drug release

Understanding the dynamics of stimuli-sensitive self-assembled nanomaterials is crucial for their potential applications, however, it is relatively difficult to quantitatively probe the dynamics using conventional analytical techniques because of the low sensitivity and selectivity of these techniques. In this research, TEMPO-containing nonamphiphilic copolymers are synthesized and highly sensitive, selective and non-destructive electron spin resonance (ESR) spectroscopy is applied to study their dynamics in stimuli-sensitive self-assembled nanostructures in different microenvironments. ESR spectroscopy reveals the structural changes of the smart supramolecular nanoparticles (SNPs) in their controlled assemblies by observing the molecular-level interactions. In order to obtain the dynamical evidence, several spectroscopic parameters such as rotational correlation time, hyperfine splitting constant, intensity increase/decrease and anisotropy parameters are determined from the ESR spectra. The stimuli-sensitivity and dynamics strongly depend on the SNPs’ microenvironment correlated to several chemical and physiological parameters such as radical concentration, pH, redox agent, temperature, polarity and viscosity. The results provide quantitative insights into the interface of the supramolecular assemblies. The nanoparticles showed good biocompatibility confirmed by the hemolysis test and the MTT assay. Furthermore, proof-of-concept experiments were carried out to demonstrate the SNPs’ stimuli-sensitive performance as a hydrophobic anticancer drug nanocarrier. This effort can be used to design other colloidal nano-platforms for the potential application in cancer treatment.

Study of factors affecting the in vitro release of diclofenac sodium from hypromelose-based gels

The aim of our study was to identify factors affecting the in vitro release of diclofenac sodium (DS) from hypromellose-based gels (HPMC).&#x0D; Materials and methods. Gels with HPMC and liquids without HPMC were studied by viscosity-rotating viscometer method and spin probe electron paramagnetic resonance spectroscopy. Rheograms were used to determine the flow behavior and the apparent viscosity, and the EPR spectra were used to determine the rotational correlation time (τ–1) of the dissolved spin probes. The in vitro release tests were performed using vertical diffusion cells according to a validated procedure. The assay of DS and isopropyl alcohol (IPA) in the receptor medium was performed by high performance liquid chromatography (HPLC) and gas chromatography (GC) according to validated procedures, and the water content was determined using semi-micro method.&#x0D; Results. The apparent viscosity of the gels increased with increasing HPMC content and depended on the HPMC grade. The high apparent viscosity of the gels did not affect the values of τ–1 of the dissolved spin probes. In viscous gels and Newtonian fluids, the composition of which corresponded to the dispersion medium of gels, the values of τ–1 were identical and were in the range of rapid rotation, which is a prerequisite for similar and rapid release of the dissolved substances from gels and liquids. It was shown that the HPMC-based gel and Newtonian liquid without HPMC in terms of in vitro release parameters DS and IPA were equivalent. During in vitro testing the release of dissolved DS increased with increasing its concentration in the gel and depended on the dispersed state of DS. When the content of IPA was changed from 45.0 % to 22.5 %, the water absorption by the gel and the release of IPA decreased, and the release of DS increased, which was due to the decrease in the solubility of DS in the gel.&#x0D; Conclusions. HPMC, which provided high apparent viscosity of the gels, did not affect the value of τ–1 of the dissolved spin probes and the in vitro release of DS from the gels. The gel and Newtonian liquid were equivalent in terms of in vitro release of DS and IPA. The release of DS altered proportionally with the concentration of DS and depended on its dispersed state. As the content of IPA decreased, the release of IPA decreased, but the release of DS increased because of the decrease in the solubility of the DS in the gel

In Situ Continuous Wave Electron Paramagnetic Resonance Investigation of the Amyloid Aggregation of Parkinson’s Protein Alpha-Synuclein—the Second Spin-Label Position

Self-aggregation of amyloid proteins is a crucial step in neurodegenerative disease. The protein alpha-synuclein (αS) is implicated in Parkinson’s disease. In an extension of the demonstration of in situ observation of intermediates in αS-aggregation by continuous wave (cw) EPR at room temperature (Zurlo et al. PLoS One 16: e0245548, 2021) by spin-label EPR, here the spin label is attached to position 90 (R1αS90), rather than at position 56. The aim is to determine, if the spin-label position affects the kinetics of aggregation and if local information on the intermediates is accessible. Probed by the MTSL ((1-Oxyl-2,2,5,5-tetramethylpyrroline-3-methyl) methanethiosulfonate) spin label at position 90, using diamagnetic dilution of 9:1 wild type αS to R1αS90, similar aggregation kinetics are found. Rotation correlation times for the spin label in the oligomer cannot be determined with sufficient accuracy to obtain local information on the oligomer under the conditions used. At the present stage, higher resolution EPR approaches, such as high-field EPR are more promising.

Fluorescent Liquid Tetrazines.

Tetrazines with branched alkoxy substituents are liquids at ambient temperature that despite the high chromophore density retain the bright orange fluorescence that is characteristic of this exceptional fluorophore. Here, we study the photophysical properties of a series of alkoxy-tetrazines in solution and as neat liquids. We also correlate the size of the alkoxy substituents with the viscosity of the liquids. We show using time-resolved spectroscopy that intersystem crossing is an important decay pathway competing with fluorescence, and that its rate is higher for 3,6-dialkoxy derivatives than for 3-chloro-6-alkoxytetrazines, explaining the higher fluorescence quantum yields for the latter. Quantum chemical calculations suggest that the difference in rate is due to the activation energy required to distort the tetrazine core such that the and the higher-lying states cross, at which point the spin-orbit coupling exceeding 10 cm allows for efficient intersystem crossing to occur. Femtosecond time-resolved anisotropy studies in solution allow us to measure a positive relationship between the alkoxy chain lengths and their rotational correlation times, and studies in the neat liquids show a fast decay of the anisotropy consistent with fast exciton migration in the neat liquid films.

Rotating-Frame Overhauser Transfer via Long-Lived Coherences

Solution-state distance restraints for protein structure determination with Ångström-level resolution rely on through-space transfer of magnetization between nuclear spins. Such magnetization transfers, named Overhauser effects, occur via dipolar magnetic couplings. We demonstrate improvements in magnetization transfer using long-lived coherences (LLCs)—singlet-triplet superpositions that are antisymmetric with respect to spin-permutation within pairs of coupled magnetic nuclei—as the magnetization source. Magnetization transfers in the presence of radio-frequency irradiation, known as ‘rotating-frame’ Overhauser effects (ROEs), are predicted by theory to improve by the use of LLCs; calculations are matched by preliminary experiments herein. The LLC-ROE transfers were compared to the transmission of magnetization via classical transverse routes. Long-lived coherences accumulate magnetization on an external third proton, K, with transfer rates that depended on the tumbling regime. I,S →K transfers in the LLC configuration for (I,S) are anticipated to match, and then overcome, the same transfer rates in the classical configuration as the molecular rotational correlation times increase. Experimentally, we measured the LLC-ROE transfer in dipeptide AlaGly between aliphatic protons in different residues K = Ala − Hα and (I,S) = Gly − Hα1,2 over a distance dK,I,S = 2.3 Å. Based on spin dynamics calculations, we anticipate that, for such distances, a superior transfer of magnetization occurs using LLC-ROE compared to classical ROE at correlation times above τC=10 ns. The LLC-ROE effect shows potential for improving structural studies of large proteins and offering constraints of increased precision for high-affinity protein-ligand complexes in slow tumbling in the liquid state.

Dissecting Monomer-Dimer Equilibrium of an RNase P Protein Provides Insight Into the Synergistic Flexibility of 5' Leader Pre-tRNA Recognition.

Ribonuclease P (RNase P) is a universal RNA-protein endonuclease that catalyzes 5’ precursor-tRNA (ptRNA) processing. The RNase P RNA plays the catalytic role in ptRNA processing; however, the RNase P protein is required for catalysis in vivo and interacts with the 5’ leader sequence. A single P RNA and a P protein form the functional RNase P holoenzyme yet dimeric forms of bacterial RNase P can interact with non-tRNA substrates and influence bacterial cell growth. Oligomeric forms of the P protein can also occur in vitro and occlude the 5’ leader ptRNA binding interface, presenting a challenge in accurately defining the substrate recognition properties. To overcome this, concentration and temperature dependent NMR studies were performed on a thermostable RNase P protein from Thermatoga maritima. NMR relaxation (R1, R2), heteronuclear NOE, and diffusion ordered spectroscopy (DOSY) experiments were analyzed, identifying a monomeric species through the determination of the diffusion coefficients (D) and rotational correlation times (τc). Experimental diffusion coefficients and τc values for the predominant monomer (2.17 ± 0.36 * 10−10 m2/s, τ c = 5.3 ns) or dimer (1.87 ± 0.40* 10−10 m2/s, τ c = 9.7 ns) protein assemblies at 45°C correlate well with calculated diffusion coefficients derived from the crystallographic P protein structure (PDB 1NZ0). The identification of a monomeric P protein conformer from relaxation data and chemical shift information enabled us to gain novel insight into the structure of the P protein, highlighting a lack of structural convergence of the N-terminus (residues 1–14) in solution. We propose that the N-terminus of the bacterial P protein is partially disordered and adopts a stable conformation in the presence of RNA. In addition, we have determined the location of the 5’ leader RNA in solution and measured the affinity of the 5’ leader RNA–P protein interaction. We show that the monomer P protein interacts with RNA at the 5’ leader binding cleft that was previously identified using X-ray crystallography. Data support a model where N-terminal protein flexibility is stabilized by holoenzyme formation and helps to accommodate the 5’ leader region of ptRNA. Taken together, local structural changes of the P protein and the 5’ leader RNA provide a means to obtain optimal substrate alignment and activation of the RNase P holoenzyme.

TRACT revisited: an algebraic solution for determining overall rotational correlation times from cross-correlated relaxation rates.

Accurate rotational correlation times (τc) are critical for quantitative analysis of fast timescale NMR dynamics. As molecular weights increase, the classic derivation of τc using transverse and longitudinal relaxation rates becomes increasingly unsuitable due to the non-trivial contribution of remote dipole-dipole interactions to longitudinal relaxation. Derivations using cross-correlated relaxation experiments, such as TRACT, overcome these limitations but are erroneously calculated in 65% of the citing literature. Herein, we developed an algebraic solutions to the Goldman relationship that facilitate rapid, point-by-point calculations for straightforward identification of appropriate spectral regions where global tumbling is likely to be dominant. The rigid-body approximation of the Goldman relationship has been previously shown to underestimate TRACT-based rotational correlation time estimates. This motivated us to develop a second algebraic solution that employs a simplified model-free spectral density function including an order parameter term that could, in principle, be set to an average backbone S2 ≈ 0.9 to further improve the accuracy of τc estimation. These solutions enabled us to explore the boundaries of the Goldman relationship as a function of the H-N internuclear distance ( r ), difference of the two principal components of the axially-symmetric 15N CSA tensor (ΔδN), and angle of the CSA tensor relative to the N-H bond vector (θ). We hope our algebraic solutions and analytical strategies will increase the accuracy and application of the TRACT experiment.

Correlated Dynamics in Ionic Liquids by Means of NMR Relaxometry: Butyltriethylammonium bis(Trifluoromethanesulfonyl)imide as an Example.

1H and 19F spin-lattice relaxation experiments have been performed for butyltriethylammonium bis(trifluoromethanesulfonyl)imide in the temperature range from 258 to 298 K and the frequency range from 10 kHz to 10 MHz. The results have thoroughly been analysed in terms of a relaxation model taking into account relaxation pathways associated with 1H–1H, 19F–19F and 1H–19F dipole–dipole interactions, rendering relative translational diffusion coefficients for the pairs of ions: cation–cation, anion–anion and cation–anion, as well as the rotational correlation time of the cation. The relevance of the 1H–19F relaxation contribution to the 1H and 19F relaxation has been demonstrated. A comparison of the diffusion coefficients has revealed correlation effects in the relative cation–anion translational movement. It has also turned out that the translational movement of the anions is faster than of cations, especially at high temperatures. Moreover, the relative cation–cation diffusion coefficients have been compared with self-diffusion coefficients obtained by means of NMR (Nuclear Magnetic Resonance) gradient diffusometry. The comparison indicates correlation effects in the relative cation–cation translational dynamics—the effects become more pronounced with decreasing temperature.

Fluorescence Depolarization Kinetics Captures Short-Range Backbone Dihedral Rotations and Long-Range Correlated Dynamics of an Intrinsically Disordered Protein.

Intrinsically disordered proteins (IDPs) do not autonomously fold into well-defined three-dimensional structures and are best described as a heterogeneous ensemble of rapidly interconverting conformers. It is challenging to elucidate their complex dynamic signatures using a single technique. In this study, we employed sensitive fluorescence depolarization kinetics by following picosecond time-resolved fluorescence anisotropy decays to directly capture the essential dynamical features of intrinsically disordered α-synuclein (α-syn) site-specifically labeled with thiol-active fluorophores. By utilizing a long-lifetime (≥10 ns) anisotropic label, we were able to discern three distinct rotational components of α-syn. The subnanosecond component represents the local wobbling-in-cone motion of the fluorophore, whereas the slower (∼1.4 ns) component corresponds to the short-range backbone dynamics governed by collective torsional fluctuations in the Ramachandran Φ-Ψ dihedral space. This backbone dihedral rotational time scale is sensitive to the local chain stiffness and slows down in the presence of an adjacent proline residue. We also observed a small-amplitude (≤10%) slower rotational correlation time (6-10 ns) that represents the long-range correlated dynamics involving a much longer segment of the polypeptide chain. These intrinsic dynamic signatures of IDPs will provide critical mechanistic underpinnings in a mosaic of biophysical phenomena involving internal friction, allosteric interactions, and phase separation.

Perchlorinated Triarylmethyl Radical 99% Enriched 13C at the Central Carbon as EPR Spin Probe Highly Sensitive to Molecular Tumbling.

Soluble stable radicals are used as spin probes and spin labels for in vitro and in vivo electron paramagnetic resonance (EPR) spectroscopy and imaging applications. We report the synthesis and characterization of a perchlorinated triarylmethyl radical enriched 99% at the central carbon, 13C1-PTMTC. The anisotropy of the hyperfine splitting with the 13C1 (Ax = 26, Ay = 25, Az = 199.5 MHz) and the g (gx = 2.0015, gy = 2.0015, gz = 2.0040) are responsible for a strong effect of the radical tumbling rate on the EPR spectrum. The rotational correlation time can be determined by spectral simulation or via the line width or the apparent Az after calibration, so the spin probe 13C1-PTMTC can be used to measure media microviscosity with high sensitivity.

Rotation of a Charged Spin Probe in Room-Temperature Ionic Liquids.

X-band electron paramagnetic resonance spectroscopy has been used to investigate the rotational diffusion of a stable, positively charged nitroxide 4-trimethylammonium-2,2,6,6-tetramethylpiperidine-1-oxyl iodide (Cat-1) in a series of 1-alkyl-3-methylimidazolium tetrafluoroborate room-temperature ionic liquids (RTILs) having alkyl chain lengths from two to eight carbons. The rotation of Cat-1 is anisotropic with the preferential axis of rotation along the NO• moiety. The Stokes-Einstein-Debye law describes the mean rotational correlation time of Cat-1, assuming that the hydrodynamic radius is smaller than the van der Waals radius of the probe. This implies that the probe rotates freely, experiencing slip boundary condition, which is solvent-dependent. The rotational correlation time of Cat-1 in RTILs can very well be fitted to a power-law functionality with a singular temperature, which suggests that the apparent activation energy of rotation exhibits non-Arrhenius behavior. Compared to the rotation of perdeuterated 2,2,6,6-tetramethyl-4-oxopiperidine-1-oxyl (pDTO), which is neutral, the rotation of Cat-1 is several times slower. The rotational anisotropy, the ratio of the rotational times of pDTO and Cat-1, and the apparent activation energy indicate the transition from a homogeneously globular structure to a spongelike structure when the alkyl chain has four carbons, which is also observed in molecular dynamics computational studies. For the first time, we have been able to show that the rotational correlation time of a solute molecule can be analyzed in terms of the Cohen-Turnbull free volume theory. The Cohen-Turnbull theory fully describes the rotation of Cat-1 in all ionic liquids in the measured temperature range.

Synergistic Enhancement of Fluorescence and Magnetic Resonance Signals Assisted by Albumin Aggregate for Dual-Modal Imaging.

Dual-modal fluorescence and magnetic resonance imaging (FLI/MRI) is important for the early diagnosis of malignant tumors. However, facile and opportune strategies to synergistically enhance fluorescence intensity and magnetic resonance (MR) contrast have rarely been reported. Herein, we present a facile strategy using albumin aggregates (AAs) to synergistically enhance the fluorescence intensity by aggregation-induced emission (AIE) and MR contrast with prolonged rotational correlation time (τR) of Gd(III) chelates and the diffusion correlation time (τD) of surrounding water molecules. The amphiphilic dual-modal FLI/MRI probe of NGd was facilely loaded into albumin pockets and then formed AAs to generate a supramolecular structure of NGd-albumin aggregates (NGd-AAs), which show excellent biocompatibility and biosafety, and exhibit superior fluorescence quantum yield and r1 over NGd with 6- and 8-fold enhancement, respectively. Moreover, compared with the clinical MRI contrast agent Gd-DOTA, r1 of NGd-AAs showed a 17-fold enhancement. Therefore, NGd-AAs successfully elicited high-performance dual-modal FLI/MRI in vitro and in vivo and high contrast MR signals were observed in the liver and tumor after intravenous injection of NGd-AAs at a dosage of 6 μmol Gd(III)/kg body weight. This generic and feasible strategy successfully realized a synergistic effect for dual-modal FLI/MRI.

Addressing the Exigent Role of a Coumarin Fluorophore toward Finding the Suitable Microenvironment of Biomimicking and Biomolecular Systems: Steering to Project the Drug Designing and Drug Delivery Study.

The photophysics of 4-azidocoumarin (4-AC), a novel fluorescent coumarin derivative, is well established by the investigation of the alteration of the microheterogeneous environment comprising two types of systems: supramolecular systems, cyclodextrins (CDs), and biomolecular systems, serum albumins (SAs). The enhanced emission of the ligand with the organized assemblies like α-CD, β-CD, and γ-CD by steady-state and time-resolved fluorescence and fluorescence anisotropy at 298 K is compared with those of bovine serum albumin (BSA) and human serum albumin (HSA). The remarkable enhancement of the emission of ligand 4-AC along with the blue shift of the emission for both the systems are visualized as the incorporation of 4-AC into the hydrophobic core of the CDs and proteins mainly due to reduction of nonradiative decay process in the hydrophobic interior of CDs and SAs. The binding constants at 298 K and the single binding site are estimated using enhanced emission and anisotropy of the bound ligand in both the systems. The marked enhancement of the fluorescence anisotropy indicates that the ligand molecule experiences a motionally constrained environment within the CDs and SAs. Rotational correlation time (θc) of the bound ligand 4-AC is calculated in both the categories of the confined environment using time-resolved anisotropy at 298 K. Molecular docking studies for both the variety of complexes of the ligand throw light to assess the location of the ligand and the microenvironment around the ligand in the ligand–CD and ligand–protein complexes. Solvent variation study of the probe 4-AC molecule in different polar protic and aprotic solvents clearly demonstrates the polarity and hydrogen-bonding ability of the solvents, which supports the alteration of the microenvironments around 4-AC due to binding with the biomimicking as well as biomolecular systems. Dynamic light scattering is employed to determine the hydrodynamic diameter of free BSA/HSA and complexes of BSA/HSA with the ligand 4-AC.

Core-Shell Tecto Dendrimers Enable Enhanced Tumor MR Imaging through an Amplified EPR Effect.

Development of nanoplatforms that can amplify the passive tumor targeting effect based on enhanced permeability and retention (EPR) effect is crucial for precision cancer nanomedicine applications. Herein, we present the development of core-shell tecto dendrimers (CSTDs) as a platform for enhanced tumor magnetic resonance (MR) imaging through an amplified EPR effect. In this work, poly(amidoamine) (PAMAM) dendrimers of generation 5 (G5) were decorated with β-cyclodextrin (CD) and then assembled with G3 PAMAM dendrimers premodified with adamantane (Ad) via supramolecular recognition of CD and Ad. The formed G5-CD/Ad-G3 CSTDs were conjugated with tetraazacyclododecane tetraacetic acid (DOTA)-Gd(III) chelators and further acetylated to neutralize the remaining CSTD periphery amines. We reveal that the formed CSTD.NHAc-DOTA(Gd) (CSTD-D-Gd) complexes have a narrow size distribution and satisfactory colloidal stability, and are cytocompatible within the concentration range studied. Compared to the single dendrimer counterpart of G5.NHAc-DOTA(Gd) (G5-D-Gd) complexes, the CSTD-D-Gd complexes with a higher molecular weight and volume possess a longer rotation correlation time, hence having a longitudinal relaxivity (r1) of 7.34 mM-1 s-1, which is 1.5 times larger than that of G5-D-Gd complexes (4.92 mM-1 s-1). More importantly, the CSTD-D-Gd complexes display better permeability in the three-dimensional (3D) cell spheroids in vitro through fluorescence imaging and a more significant EPR effect for improved tumor MR imaging in vivo than the G5-DOTA-Gd complexes. The generated CSTD-D-Gd complexes may be adopted for enhanced tumor MR imaging through an amplified passive EPR effect and also be further extended for different cancer theranostic applications.