Amyloid-mimicking toxic nanofibers generated via self-assembly of dopamine.

Molecular self-assembly of biologically relevant aromatic metabolites is known to generate cytotoxic nanostructures and this unique property has opened up new concepts in the molecular mechanisms of metabolite-linked disorders. Because aromaticity is intrinsic to the chemical structure of some important neuromodulators, the question of whether this property can promote their self-assembly into toxic higher order structures is highly relevant to the advancement of both fundamental and applied research. We show here that dopamine, an aromatic neuromodulator of high significance, undergoes self-assembly, under physiological buffer conditions, yielding cytotoxic supramolecular nanostructures. The oxidation of dopamine seems crucial in driving the self-assembly, and substantial inhibition effect was observed in the presence of antioxidants and acidic buffers. Strong H-bonds and π-π interactions between optimally-oriented dopamine molecules were found to stabilize the dopamine nanostructure which displayed characteristic ß-structure-patterns with hydrophobic exterior and hydrophilic interior moieties. Furthermore, dopamine nanostructures were found to be highly toxic to human neuroblastoma cells, revealing apoptosis and necrosis-mediated cytotoxicity. Abnormal fluctuation in the dopamine concentration is known to predispose a multitude of neuronal complications, hence, the new findings of this study on oxidation-driven buildup of amyloid-mimicking neurotoxic dopamine assemblies may have direct relevance to the molecular origin of several dopamine related disorders.

Genesis of Neurotoxic Hybrid Nanofibers from the Coassembly of Aromatic Amino Acids.

Considering the relevance of accumulation and self-assembly of metabolites and aftermath of biological consequences, it is important to know whether they undergo coassembly and what properties the resultant hybrid higher-order structures would exhibit. This work reveals the inherent tendency of aromatic amino acids to undergo a spontaneous coassembly process under physiologically mimicked conditions, which yields neurotoxic hybrid nanofibers. Resultant hybrid nanostructures resembled the ß-structured conformers stabilized by H-bonds and π-π stacking interactions, which were highly toxic to human neuroblastoma cells. The hybrid nanofibers also showed strong cross-seeding potential that triggered in vitro aggregation of diverse globular proteins and brain extract components, converting the native structures into cross-ß-rich amyloid aggregates. The heterogenic nature of the hybrid nanofibers seems crucial for their higher toxicity and faster cross-seeding potential as compared to the homogeneous amino acid nanofibers. Our findings reveal the importance of aromaticity-driven optimized intermolecular arrangements for the coassembly of aromatic amino acids, and the results may provide important clues to the fundamental understanding of metabolite accumulation-related complications.

Dilution-Induced Physico-Chemical Changes of Metal Oxide Nanoparticles Due to Homeopathic Preparation Steps of Trituration and Succussion.

BACKGROUND: Although the presence of starting materials in extreme dilutions of homeopathic medicines has been established, the physico-chemical changes of these materials induced by the manufacturing steps-that is, solid-solid mixing involving grinding (trituration) and slurry mixing involving impact (succussion), followed by dilution-are still unknown. METHODS: We subjected cupric oxide and zinc oxide nanoparticles (NPs) to the homeopathic processes of trituration and succussion, followed by dilution up to 6 cH. Particle image velocimetry was employed to analyze the fluid motion during succussion and its effect on the NPs. The resulting microstructural and chemical changes at different dilution steps were determined by X-ray photoelectron spectroscopy, Fourier-transform infrared spectroscopy and transmission electron microscopy. RESULTS: The succussion triggered multi-sized bubble generation and turbulent fluid motion up to a duration of 400 ms, with maximum average velocity of 0.23 m/s. Due to 1% transfer of kinetic energy from a moving eddy with this velocity, upon collision, the rate of temperature change in a particle of size 1 µm and 1 nm was predicted to rise by approximately 102 K/s and 106 K/s respectively. During trituration, the oxide NPs reduced to metals and did not aggregate by remaining within lactose, but they converted to oxidized finer NPs after impact. Silicate chains leached from the vial cross-linked after third dilution, forming large macro-particles and encapsulating the NPs that were retained and carried at higher dilution steps. CONCLUSION: The results showed that the NPs sustained significant rate of temperature change due to energy transfer from moving eddies during succussion. Different physico-chemical changes, such as size reduction, successive reduction and oxidation of NPs, and morphological changes, were achieved through trituration and succussion. The retention of NPs within cross-linked poly-siloxane chains reveals the importance of both the borosilicate glass vial and the ethanol solution during preparation of homeopathic medicines.

Myricetin inhibits amyloid fibril formation of globular proteins by stabilizing the native structures.

Myricetin has been identified as a naturally occurring flavonoid class of polyphenolic compound which shows multiple medical benefits including antidiabetic, anticancerous and antioxidant properties. Here, we report the protective effect of myricetin against in vitro amyloid fibril formation of selected globular proteins. The results reveal that myricetin is capable of inhibiting amyloid fibril formation of both insulin and serum albumin. Seed-induced aggregation of both proteins was also substantially suppressed in the presence of myricetin. Fluorescence quenching data indicated binding of myricetin with protein monomers as well as fibrils. The molecular docking studies revealed strong affinity of myricetin for both the native and partially unfolded conformation of proteins mediated by H-bonds and hydrophobic interactions. Myricetin was also observed to promote disassembly of mature amyloid fibrils. The results reveal that myricetin molecule has the potential for suppressing amyloid formation and such an inherent property may help in developing myricetin-based antiamyloid drugs.

A biodegradable fluorescent nanohybrid for photo-driven tumor diagnosis and tumor growth inhibition.

Specific targeting and phototriggered therapy in mouse model have recently emerged as the starting point of cancer theragnosis. Herein, we report a bioresponsive and degradable nanohybrid, a liposomal nanohybrid decorated with red emissive carbon dots, for localized tumor imaging and light-mediated tumor growth inhibition. Unsaturated carbon dots (C-dots) anchored to liposomes convert near-infrared (NIR) light into heat and also produce reactive oxygen species (ROS), demonstrating the capability of phototriggered cancer cell death and tumor regression. The photothermal and oxidative damage of breast tumor by the nonmetallic nanohybrid has also been demonstrated. Designed nanoparticles show excellent aqueous dispersibility, biocompatibility, light irradiated enhanced cellular uptake, release of reactive oxygen species, prolonged and specific tumor binding ability and good photothermal response (62 °C in 5 minutes). Safe and localized irradiation of 808 nm light demonstrates significant tumor growth inhibition and bioresponsive degradation of the fluorescent nanohybrid without affecting the surrounding healthy tissues.

Bioconductive 3D nano-composite constructs with tunable elasticity to initiate stem cell growth and induce bone mineralization.

Bioactive 3D composites play an important role in advanced biomaterial design to provide molecular coupling and improve integrity with the cellular environment of the native bone. In the present study, a hybrid lyophilized polymer composite blend of anionic charged sodium salt of carboxymethyl chitin and gelatin (CMChNa-GEL) reinforced with nano-rod agglomerated hydroxyapatite (nHA) has been developed with enhanced biocompatibility and tunable elasticity. The scaffolds have an open, uniform and interconnected porous structure with an average pore diameter of 157±30µm and 89.47+0.03% with four dimensional X-ray. The aspect ratio of ellipsoidal pores decrease from 4.4 to 1.2 with increase in gelatin concentration; and from 2.14 to 1.93 with decrease in gelling temperature. The samples were resilient with elastic stain at 1.2MPa of stress also decreased from 0.33 to 0.23 with increase in gelatin concentration. The crosslinker HMDI (hexamethylene diisocyanate) yielded more resilient samples at 1.2MPa in comparison to glutaraldehyde. Increased crosslinking time from 2 to 4h in continuous compression cycle show no improvement in maximum elastic stain of 1.2MPa stress. This surface elasticity of the scaffold enables the capacity of these materials for adherent self renewal and cultivation of the NTERA-2 cL.D1 (NT2/D1), pluripotent embryonal carcinoma cell with biomechanical surface, as is shown here. Proliferation with MG-63, ALP activity and Alizarin red mineralization assay on optimized scaffold demonstrated ***p<0.001 between different time points thus showing its potential for bone healing. In pre-clinical study histological bone response of the scaffold construct displayed improved activity of bone regeneration in comparison to self healing of control groups (sham) up to week 07 after implantation in rabbit tibia critical-size defect. Therefore, this nHA-CMChNa-GEL scaffold composite exhibits inherent and efficient physicochemical, mechanical and biological characteristics based on gel concentrations, gelatin mixing and gelling temperature thus points to creating bioactive 3D scaffolds with tunable elasticity for orthopedic applications.

Establishing the interfacial nano-structure and elemental composition of homeopathic medicines based on inorganic salts: a scientific approach.

Extremely dilute systems arise in homeopathy, which uses dilution factors 10(60), 10(400) and also higher. These amounts to potencies of 30c, 200c or more, those are far beyond Avogadro's number. There is extreme skepticism among scientists about the possibility of presence of starting materials due to these high dilutions. This has led modern scientists to believe homeopathy may be at its best a placebo effect. However, our recent studies on 30c and 200c metal based homeopathic medicines clearly revealed the presence of nanoparticles of starting metals, which were found to be retained due to the manufacturing processes involved, as published earlier.(9,10) Here, we use HR-TEM and STEM techniques to study medicines arising from inorganic salts as starting materials. We show that the inorganic starting materials are present as nano-scale particles in the medicines even at 1 M potency (having a large dilution factor of 10(2000)). Thus this study has extended our physicochemical studies of metal based medicines to inorganic based medicines, and also to higher dilution. Further, we show that the particles develop a coat of silica: these particles were seen embedded in a meso-microporous silicate layer through interfacial encapsulation. Similar silicate coatings were also seen in metal based medicines. Thus, metal and inorganic salt based homeopathic medicines retain the starting material as nanoparticles encapsulated within a silicate coating. On the basis of these studies, we propose a universal microstructural hypothesis that all types of homeopathic medicines consist of silicate coated nano-structures dispersed in the solvent.

Evidence of rapid coaggregation of globular proteins during amyloid formation.

The question of how an aggregating protein can influence aggregation of other proteins located in its vicinity is particularly significant because many proteins coexist in cells. We demonstrate in vitro coaggregation and cross-seeding of lysozyme, bovine serum albumin, insulin, and cytochrome c during their amyloid formation. The coaggregation process seems to be more dependent on the temperature-induced intermediate species of these proteins and less dependent on their sequence identities. Because amyloid-linked inclusions and plaques are recognized as multicomponent entities originating from aggregation of the associated protein, these findings may add new insights into the mechanistic understanding of amyloid-related pathologies.

Chain length dependence of polyol synthesis of zinc ferrite nanoparticles: why is diethylene glycol so different?

Superparamagnetic ZnFe2O4 nanoparticles with size range of 28-38 nm were synthesized by polyol process based on use of varying chain length glycols as solvent. We have offered, for the first time, the plausible mechanism behind in situ formation of zinc ferric oxalate hydroxide hydrate [Fe2Zn(C2O4)2(OH)3](+)·4H2O complex from diethylene and polyethylene glycol. We are also reporting, the magnetic properties of above complexes. We have found a ferromagnetic ordering in precursor complex compounds. The intermediate hydrocarbon chain between the oxalato bridged metal cations plays a crucial role in obtaining anomalous magnetic behavior. ZnFe2O4 nanoparticles obtained after annealing the DEGylated precursor complex (precursor complex formed in diethylene glycol) showed the highest superparamagnetic (SPM) behavior (22.4 emu g(-1)) among others. The reasons for anomalous SPM behavior of ZnFe2O4 nanoparticles are explained on the basis of the degree of inversion of the spinel structure, high surface-to-volume ratio, which causes non-collinear spin arrangement in a surface layer and higher oxygen concentration on the surface of dead organic layer, which increases the unpaired valence electrons leading to uncompensated surface spins.

A case study: Te in ZnSe and Mn-doped ZnSe quantum dots.

Photoluminescence (PL) behavior of ZnSe(1-y)Te(y) quantum dots is investigated by varying Te concentration as well as size. The striking effect of quantum confinement is the observation of isoelectronic center-related emission at room temperature in lieu of near-band-edge emission that dominates the optical scenario. ZnSe(0.99)Te(0.01) quantum dots were also doped by Mn(2+) ions. The Mn(2+) ion-related d-d transition is drastically suppressed by Te isoelectronic centers. Incorporation of Mn(2+) at substitutional sites in ZnSe(0.99)Te(0.01) quantum dots is also confirmed by the electron paramagnetic resonance measurements. Effect of Te isoelectronic impurity on the emission behavior is more pronounced than that of Mn(2+) ions. A subtle blueshift in the orange d-d transition is a sign of a decrease in crystal field strength. PL and photoluminescence excitation measurements on Zn(1-x)Se(0.99)Te(0.01)Mn(x) quantum dots indicate that the transition probability from the lowest unoccupied molecular orbital to Te levels is substantially larger than that to Mn(2+) d-d levels.