Nano-Osmolyte Conjugation: Tailoring the Osmolyte-Protein Interactions at the Nanoscale.

Osmolytes are small organic compounds accumulated at higher concentrations in the cell under various stress conditions like high temperature, high salt, high pressure, etc. Osmolytes mainly include four major classes of compounds including sugars, polyols, methylamines, and amino acids and their derivatives. In addition to their ability to maintain protein stability and folding, these osmolytes, also termed as chemical chaperones, can prevent protein misfolding and aggregation. Although being efficient protein folders and stabilizers, these osmolytes exhibit certain unavoidable limitations such as nearly molar concentrations of osmolytes being required for their effect, which is quite difficult to achieve inside a cell or in the extracellular matrix due to nonspecificity and limited permeability of the blood-brain barrier system and reduced bioavailability. These limitations can be overcome to a certain extent by using smart delivery platforms for the targeted delivery of osmolytes to the site of action. In this context, osmolyte-functionalized nanoparticles, termed nano-osmolytes, enhance the protein stabilization and chaperone efficiency of osmolytes up to 105 times in certain cases. For example, sugars, polyols, and amino acid functionalized based nano-osmolytes have shown tremendous potential in preventing protein aggregation. The enhanced potential of nano-osmolytes can be attributed to their high specificity at low concentrations, high tunability, amphiphilicity, multivalent complex formation, and efficient drug delivery system. Keeping in view the promising potential of nano-osmolytes conjugation in tailoring the osmolyte-protein interactions, as compared to their molecular forms, the present review summarizes the recent advancements of the nano-osmolytes that enhance the protein stability/folding efficiency and ability to act as artificial chaperones with increased potential to prevent protein misfolding disorders. Some of the potential nano-osmolyte aggregation inhibitors have been highlighted for large-scale screening with future applications in aggregation disorders. The synthesis of nano-osmolytes by numerous approaches and future perspectives are also highlighted.

Investigation on the Linear and Nonlinear Properties of Morin in Presence of Reverse Micelle and Different Oil Content in Reverse Micelle.

We investigate the linear and nonlinear optical property of Morin (MN) at different concentration (1 × 10-6 and 5 × 10-6 M) within AOT reversed micelle prepared by water-in-decane microemulsion having a constant molar ratio of water-to-surfactant molecules of 40 (W = [H2O]/[AOT] = 40) as well as the function of mass fraction of nano-droplet (MFD) values of 0.01,0.04, 0.07, and 0.1 by using UV-Visible, Fluorescence, FTIR, and Z-scan techniques. The steady-state measurement indicates that the presence of microenvironment can greatly affect the tautomeric structure of morin and also Morin property in microenvironment depends upon the amount of oil and Morin concentration. The increase in dipole moment from the ground state to excited state in microenvironment indicate the change in the molecular structure on morin. Morin does not show any nonlinear absorption property but the nonlinear refractive index is observed as a function of Morin concentration as well as MFD values which are due to the thermal agitation of formed dimers. Morin nonlinearity.

Metal oxide QD based ultrasensitive microsphere fluorescent sensor for copper, chromium and iron ions in water.

Herein we developed a rapid, cheap, and water-soluble ultra-sensitive ZnO quantum dot (QD) based metal sensor for detecting different hazardous metal ions up to the picomolar range in water. Various spectroscopic and microscopic techniques confirmed the formation of 2.15 ± 0.46 µm of ZnO QD conjugated CMC microspheres (ZCM microspheres) which contain 5.5 ± 0.5 nm fluorescent zinc oxide (ZnO) QDs. Our system, as a promising sensor, exhibited excellent photostability and affinity towards various heavy metal ions. The detection limits were calculated to be 16 pM for Cu2+ and 0.18 nM for Cr6+ ions which are better than previously reported values. The simple fluorescence 'turn off' property of our ZCM microsphere sensor system can serve a two-in-one purpose by not only detecting the heavy metals but also quantifying them. Nonetheless, pattern recognition for different heavy metals helped us to detect and identify multiple heavy metal ions. Finally, their practical applications on real samples also demonstrated that the ZCM sensor can be effectively utilized for detection of Cr6+, Fe3+, Cu2+ present in the real water samples. This study may inspire future research and design of target fluorescent metal oxide QDs with specific functions.

Probing the reverse micelle environment with a cationic dye by varying oil and water content of micelles.

In the present work, spectroscopic properties of crystal violet (CV) within water-in-hexane sodium base (2-ethyl hexyl) sulfosuccinate (AOT) reverse micellar environment was investigated as a function of water contents (W = [H2O]/[AOT] = 6,10) in AOT and also function of mass fraction of nanodroplet (MFD = 0.01, 0.04, 0.07, and 0.1) by using UV/visible, and fluorescence techniques. A deviation in the absorption spectra of crystal violet at the high CV concentration (0.002 M) was observed in the AOT reversed micelles. The Quenching in the emission intensity of crystal violet at high CV concentration and blue shift in λmax of fluorescence of dye was observed as a function of MFD in AOT RMs. At high concentration of dye, molecules of CV can be reside interfacial region but near core of AOT. At low dye concentration, the CV molecules also can be in interfacial region but far from core. The Stokes shift of CV at high concentration decreased with mass fraction of nano-droplet (MFD), whereas at the low CV concentration, its variation often increased. The ratio of ground state to the excited state dipole moment of crystal violet dye also affected by the change in W values and MFD values. The change in dipole moment and stokes shift indicates the different environment around CV for different W and MFD values.

Sensing of Different Human Telomeric G-Quadruplex DNA Topologies by Natural Alkaloid Allocryptopine Using Spectroscopic Techniques.

This article describes how a natural alkaloid allocryptopine (ALL) is able to differentiate two forms of biologically relevant human telomeric (htel22) G-quadruplex DNAs (GQ-DNA) depending on the presence of K+ and Na+ ions by steady-state and time-resolved spectroscopic techniques. For both interactions, predominant involvements of static-type quenching mechanism with the negligible influence of dynamic collision are established by UV-vis absorption and fluorescence emission study, which is further supported by fluorescence lifetime measurements. ALL exhibits appreciable affinity toward both GQ-DNAs. Both the mixed-hybrid (3 + 1) quadruplex structures in K+ ions and the basket-type antiparallel quadruplex structure under Na+ condition are converted to parallel types in the presence of ALL. Fluorescence intercalator displacement assay experiment revealed modest selectivity of ALL to both quadruplexes over duplex DNA along with higher selectivity for antiparallel types among the two quadruplexes via groove and/or loop binding, which is distinct from the conventional π-stacking of the ligands on external G-quartets. ALL stabilized both GQ-DNA topologies moderately. The differences in the dynamics of ALL within both DNA environments have been demonstrated vividly by time-resolved anisotropy measurements using the wobbling-in-cone model. These results suggest groove binding with antiparallel G-quartet with high affinity and moderate loop binding with mixed-hybrid G-quartet accompanied by the partial end stacking additionally in both of the cases. Our conclusions are further supported by steady-state anisotropy measurements and molecular docking. The present investigation can be used in the development of a biocompatible antitumour/anticancer agent targeting particular GQ-DNA conformation.

Nanosensing of Pesticides by Zinc Oxide Quantum Dot: An Optical and Electrochemical Approach for the Detection of Pesticides in Water.

Present study reveals the low concentrations (∼4 ppm) of pesticide sensing vis-à-vis degradation of pesticides with the help of nontoxic zinc oxide quantum dots (QD). In our study, we have taken four different pesticides viz., aldrin, tetradifon, glyphosate, and atrazine, which are widely used in agriculture and have structural dissimilarities/diversity. By using optical sensing techniques such as steady state and time-resolved fluorescence, we have analyzed the detailed exciton dynamics of QD in the presence of different pesticides. It has been found that the pesticide containing good leaving groups (-Cl) can interact with QD promptly and has high binding affinity (∼107 M-1). The different binding signatures of QD with different pesticides enable us to differentiate between the pesticides. Time resolved fluorescence spectroscopy provides significant variance (∼150-300 ns) for different pesticides. Furthermore, a large variation (105 Ω to 7 × 104 Ω) in the resistance of QD in the presence of different pesticides was revealed by electrochemical sensing technique. Moreover, during the interaction with pesticides, QD can also act as a photocatalyst to degrade pesticides. Present investigation explored the fact that the rate of degradation is positively affected by the binding affinity, i.e., the greater the binding, the greater is the degradation. What is more, both optical and electrochemical measurements of QD, in tandem, as described in our study could be utilized as the pattern recognition sensor for detection of several pesticides.

Quantitative measurement of fluorescence brightness of single molecules.

Single-molecule fluorescence spectroscopy and imaging probe many characteristics of the fluorescence from individual molecules like relative intensity, polarization, lifetime and spectrum. However, such an important and fundamental parameter as absolute fluorescence intensity (or in other words fluorescence brightness), which is proportional to the absorption cross section and fluorescence quantum yield, has not yet been sufficiently exploited in the field. One reason for that is the difficulty of absolute fluorescence brightness measurements. In the present work a detailed description of fluorescence brightness measurements of single molecules is given. We discuss several important factors like the power density and polarization of excitation light, the substrates and the local environment. It is shown that the fluorescence brightness of a single molecule indeed can be measured with sufficient accuracy and used as a powerful parameter for characterization of materials at single molecule/particle level. The brightness of a single object can give similar information as the fluorescence quantum yield that is crucial for understanding the photophysical properties for individual multi-chromophoric systems in inhomogeneous environments.

Manipulating energy transfer in copolymer-based nanocomposites by their controlled nanocaging and release of an ionic styryl dye: a case of an ultrasensitive pH sensor.

We report an interesting pH-tunable energy transfer between an acceptor ionic styryl dye 2-(4-(dimethylamino)styryl)-1-methylpyridinium iodide and a donor charge-transfer dye 1,8-naphthalimide in a vesicular medium. The polyethylene-b-polyethylene glycol block copolymer intercalates with the sodium dodecyl sulfate anionic surfactant to form self-aggregated nanocomposites. These nanocomposites interact with the donor molecules in aqueous solution to form "vesicles", and the donor molecules become attached on the outer wall by hydrogen bonding. The acceptor molecules are observed to be loaded in the vesicular interior. By controlling the spectral overlap of the donor and acceptor molecules by changing the pH of the medium, the energy-transfer efficiency in vesicles has been studied. The efficiency of energy transfer in vesicular media (55%) is found to be less compared to that in aqueous media (80%) at pH 7. The fall in efficiency has been attributed to the perturbation imparted by the vesicular wall due to the good matching of the donor-acceptor distance with the wall thickness. At low pH, the efficiency shows an abrupt increase (95%) due to the release of the acceptor molecules from the vesicular medium causing subsequent reduction of donor-acceptor separation and an increase of the spectral overlap at that pH.

Spectral signature of 2-[4-(dimethylamino)styryl]-1-methylquinolinium iodide: a case of negative solvatochromism in water.

Photophysics of the 2-[4-(dimethylamino) styryl]-1-methylquinolinium iodide (DASQMI) molecule has been studied in different solvents by steady-state and time-resolved emission spectroscopy and also with quantum chemical calculations. The probe molecule exhibits a strong solvent-polarity-dependent characteristic. The low-energy fluorescence band of DASQMI shows an anomalous 40 nm blue shift in water from that in dimethyl sulfoxide (DMSO); though in deuterium oxide the normal trend of red shift was observed. A marked increase in intensity of this band at 77 K and an increase in lifetime in viscous solvent point clearly to the intramolecular charge-transfer (ICT) character of the low-energy band. From the temperature-dependent emission and emission spectra in mixed solvents, the negative solvatochromism of DASQMI has been established, which means that the ICT state moves toward ground state with polarity and hydrogen-bond ability and beyond a critical dielectric constant coupled with protic nature of the solvent ground state gets further stabilized to show anomalous blue shift. In ethanol, below a critical temperature, 253 K, a blue shift starts due to greater solvent molecular polarization. A third long-lifetime component with dominant 75% amplitude was observed only in aqueous solution and may be due to the cis-isomer of hydrophobic DASQMI, a stable form in the excited state predicted from polarizable continuum model (PCM) calculations in water with 6-31G+(d,p) as basis set.

Reverse micelle induced flipping of binding site and efficiency of albumin protein with an ionic styryl dye.

The effect of reverse micelle environment on the binding mechanism of 2-(4-(dimethylamino)styryl)-1-methylpyridinium iodide (DASPMI) with Bovine Serum Albumin (BSA) compared with that in buffer solution has been investigated in this paper with the help of steady state and time-resolved emission spectroscopy along with molecular docking to have a correct picture about binding. The binding of DASPMI with attachment efficiency of 30% and 70% at site I (subdomain IIA) and site II (subdomain IIIA) of BSA, respectively, in buffer solution gets reversed inside a reverse micelle. The bigger cavity size of site II in buffer solution ushers the dye with increased attachment efficiency and in reverse micelle change in pi-stacking and hydrophobic interaction control the attachment efficiency. The calculated Forster distance gets curtailed as the environment changes from buffer to reverse micelle. The binding becomes stronger with a smaller gap between the probe and Trp-214 inside the reverse micelle than that in buffer solution.

Influence of surfactants on the excited state photophysics of 4-nitro-1-hydroxy-2-naphthoic acid.

This paper reports the changed dynamics of excited state proton transfer (ESPT) photophysics of 4-nitro-1-hydroxy-2-naphthoic acid (NHNA) in ionic and nonionic micelles, as revealed from time-resolved emission spectra (TRES) and time-resolved area normalized emission spectra (TRANES). Both the neutral and anionic bands of NHNA are enhanced and blue-shifted inside the micelles. Quenching of emission by metal Cu(2+) reveals that NHNA does not penetrate into the core of the micelles; rather it binds to the micelle/water interfacial region. The fast decay component is due to decay of normal species and the slow component is assigned to decay of anionic species. In all the micelles the decay time becomes longer by three times compared to that in water. The efficiency of formation of anionic state from the locally excited state gets enhanced in micellar media due to enhanced sojourn in the first excited state in increased viscous medium around NHNA. TRANES data also indicate that of the normal and anionic species in the aqueous phase the anionic species increases in the micelles as the time evolved.

Quest for mode of binding of 2-(4-(dimethylamino)styryl)-1-methylpyridinium iodide with calf thymus DNA.

The mode of binding of 2-(4-(dimethylamino)styryl)-1-methylpyridinium iodide (DASPMI) with calf thymus DNA as revealed from different steady state and time-resolved emission spectroscopic measurements has been reported in this paper. Fluorescence enhancement of DASPMI and its quenching by potassium iodide (KI) points to groove binding of dye with ct-DNA, rather than intercalation in the ct-DNA helix. An increase in steady state anisotropy and fluorescence lifetime hints at binding with ct-DNA. The value of binding constant from emission and association constant from circular dichroic spectrum also indicates weak binding. The strong dependence on ionic strength or salt in controlling the binding of DASPMI with ct-DNA by electrostatic interaction confirms groove binding. The high semicone angle of DASPMI in ct-DNA certainly rules out the possibility of intercalated bonding. A theoretical modeling shows that the probe is bound to ct-DNA as a crescent with a curvature of 11.35 A, which is the previously known curvature of probe in the minor groove.

On the spectral behavior of an ionic styryl dye: effect of micelle-polyethylene-block-polyethylene glycol diblock copolymer assembly.

The interaction of anionic micelle sodium dodecyl sulfate (SDS) and amphiphilic block copolymers polyethylene-b-polyethylene glycol (PE-b-PEG) and the sharp change of excited-state charge-transfer complex photophysics of 2-(4-(dimethylamino)styryl)-1-methylpyridinium iodide (DASPMI) inside of the supramolecular assembly have been addressed in the paper. The dramatic enhancement of emission intensity of DASPMI incorporated inside of the nanostructure formed by micellar and polymeric chains indicates a completely different environment compared to that in the water and micellar system. A huge increase in the rotational relaxation time obtained from time-resolved anisotropy decay and the value of the order parameter is indicative of a very restrictive regime in the self-assembly system. The wobbling and translational motion of the probe is also restricted inside of the micelle-polymer aggregate due to the presence of polymer chains. The translational diffusion coefficient is drastically reduced due to the aggregation.