Effects of Palladium(II) Chlorocomplex Speciation on the Controlled Interaction with a Polyaniline Film in Acid.

In this study, we determined the Pd(II) chlorocomplex species that has the most favorable interaction with an electropolymerized and protonated polyaniline (PANI) film. This study was completed with the intent to use this species to electrochemically build atomic palladium clusters in the PANI matrix. Varying amounts of NaCl were added to a K2PdCl4/HClO4 solution to result in three species studied: PdCl2(H2O)2, PdCl3(H2O)-, and PdCl42-. UV-vis spectroscopy was used to confirm the speciation, and Raman spectroscopy, X-ray photoelectron spectroscopy, and cyclic voltammograms were used to probe the interaction between the Pd species and PANI. It was determined that PdCl3(H2O)- most effectively interacts with PANI as a result of the charge balance between the anion and the protonated nitrogen-containing groups in the polymer. It has been also found that some fraction of inserted Pd(II) cannot be reduced to Pd(0).

Hyperoxia Induces Intracellular Acidification in Neonatal Mouse Lung Fibroblasts: Real-Time Investigation Using Plasmonically Enhanced Raman Spectroscopy.

It is important to understand the molecular mechanisms underlying oxygen toxicity, which contributes to multiple human disorders. The archetype model of oxygen toxicity is neonatal lung injury induced by hyperoxia exposure. Here, we utilized plasmonically enhanced Raman spectroscopy (PERS) in combination with fluorescence and proteomic analysis to provide comprehensive information on hyperoxia-induced biomolecular modifications in neonatal mouse lung fibroblasts (nMLFs). During this study, we made the novel observation that hyperoxia induces intracellular acidification in nMLF, which we probed in real-time using label-free PERS. We found that intracellular acidification induces conformational modifications in proteins followed by significant changes in Raman vibrations corresponding to aromatic amino acids such as phenylalanine and tryptophan as well as cysteine moieties. Hyperoxia-induced intracellular pH changes and subsequent modifications in protein expression and associated post-translational modifications within the cells were further validated by fluorescence and proteomic analysis. These new insights may help identifying unique oxidant stress-induced mechanisms in disease processes and may guide the development of more efficient therapeutic strategies.

Lewis Acid Based Sorption of Trace Amounts of RuCl3 by Polyaniline.

A sorption process of RuCl3 in phosphate buffer by polyaniline (PANI) powder chemically synthesized from phosphoric acid was spectrophotometrically monitored as a function of time. It was determined that the sorption process follows the Langmuir and Freundlich isotherms, and their constants were evaluated. It was determined that chemisorption was the rate-controlling step. By conducting detailed studies, we assigned the chemisorption to Lewis acid based interactions of the sorbent electron pair localized at the benzenoid amine (-NH2) and quinoid imine (═NH) groups, with the sorbate, RuCl3, as the electron acceptor. The stability of the interaction over a period of ∼1 week showed that the presence of the Ru(III) in the PANI matrix reverses its state from emeraldine base to emeraldine salt, resulting in a change of conductivity. The partial electron donor based charge transfer is a slow process as compared to the sorption process involving Brønsted acid doping.

Unraveling the biomolecular snapshots of mitosis in healthy and cancer cells using plasmonically-enhanced Raman spectroscopy.

Owing to the dynamic and complex nature of mitosis, precise and timely executions of biomolecular events are critical for high fidelity cell division. In this context, visualization of such complex events at the molecular level can provide vital information on the biomolecular processes in abnormal cells. Here, we explored the plasmonically enhanced light scattering properties of functionalized gold nanocubes (AuNCs) together with surface-enhanced Raman spectroscopy (SERS) to unravel the complex and dynamic biological processes involved in mitosis of healthy and cancerous cells from its molecular perspectives. By monitoring various stages of mitosis using SERS, we noticed that relatively high rate of conversion of mitotic proteins from their α-helix structure to ß-sheet conformation is likely in the cancer cells during meta-, ana-, and telophases. Unique biochemical modifications to the lipid and amino acid moieties, associated with the observed protein conformational modifications, were also identified. However, in healthy cells, the existence of proteins in their ß conformation was momentary and was largely in the α-helix form. The role of abnormal conformational modifications of mitotic proteins on the development of anomalous mitotic activities was further confirmed by looking at plasmonic nanoparticle-induced cytokinesis failure in cancer cells. Our findings illustrate the vast possibilities of SERS in real-time tracking of complex, subtle, and momentary modifications of biomolecules in live cells, which could provide new insights to the role of protein conformation dynamics during mitosis on the development of cancer and many other diseases.

Detection of target ssDNA using a microfabricated Hall magnetometer with correlated optical readout.

Sensing biological agents at the genomic level, while enhancing the response time for biodetection over commonly used, optics-based techniques such as nucleic acid microarrays or enzyme-linked immunosorbent assays (ELISAs), is an important criterion for new biosensors. Here, we describe the successful detection of a 35-base, single-strand nucleic acid target by Hall-based magnetic transduction as a mimic for pathogenic DNA target detection. The detection platform has low background, large signal amplification following target binding and can discriminate a single, 350 nm superparamagnetic bead labeled with DNA. Detection of the target sequence was demonstrated at 364 pM (<2 target DNA strands per bead) target DNA in the presence of 36 µM nontarget (noncomplementary) DNA (<10 ppm target DNA) using optical microscopy detection on a GaAs Hall mimic. The use of Hall magnetometers as magnetic transduction biosensors holds promise for multiplexing applications that can greatly improve point-of-care (POC) diagnostics and subsequent medical care.

Lipid bilayer discs and banded tubules: photoinduced lipid sorting in ternary mixtures.

The self-assembly of biological amphiphiles has proved a fascinating topic in recent years, the hollow cylindrical lipid tubule morphology being of particular interest due to its potential applications in "soft" microtechnologies. Lateral coexistence of liquid-ordered (lo) and liquid-disordered (ld) phases, which may resemble raft formation in cell membranes, was investigated in lipid tubules, prepared from 1,2-dioleoyl-sn-glycero-3-phosphocholine, egg-sphingomyelin, and cholesterol. Fluorescence microscopy shows that the appearance of micrometer-scale lo domains in the lipid tubule is not an intrinsic phase behavior of the system but a consequence of photoinduced lipid peroxidation. Most interestingly, new photoinduced bilayer structures: lipid discs, essentially stable flattened liposomes, were observed for the first time in a model membrane system. This investigation not only aids in our understanding of lipid sorting phenomena in cell membranes but also demonstrates how control of this process may provide a route to the generation of new, functional structures.

Elucidation of ultraviolet radiation-induced cell responses and intracellular biomolecular dynamics in mammalian cells using surface-enhanced Raman spectroscopy.

Fingerprinting biochemical changes associated with cellular responses to external stimuli can provide vital information on the dynamics of biological processes and their defense mechanisms. In this study, surface-enhanced Raman spectroscopy (SERS) has been used to elucidate biomolecular dynamics on the response of healthy and cancerous cells towards ultraviolet (UV) light irradiation at the cellular level in real-time. We have identified a number of physiochemical damages to proteins, especially to the chemical structure of the sulfur and aromatic amino acid containing moieties, as well as changes in secondary structure. Furthermore, we found that continuous exposure of short wave UV-C light (254 nm) to living cells can photolytically damage intracellular proteins and can completely arrest nanoparticle transport and trigger apoptosis. However, under similar conditions, this was not observed when the cells were exposed to long wave UV-A light (365 nm). These biomolecular events were probed in real-time using SERS and dark-field (DF) imaging. Specifically, this technique has been utilized for the real-time evaluation of a unique cellular defense mechanism in cancer cells towards UV exposure. Our technique provides a powerful approach to understand the mechanisms of UV light-triggered cell death, protein dynamics, and enhanced cell repair and defense machinery within cancer cells through actively monitoring molecular vibrations.

Microcapsules with three orthogonal reactive sites.

Polymeric microcapsules containing reactive sites on the shell surface and two orthogonally reactive polymers encapsulated within the interior are selectively labeled. The capsules provide three spatially separate and differentially reactive sites. Confocal fluorescence microscopy is used to characterize the distribution of labels. Polymers encapsulated are distributed homogeneously within the core and do not interact with the shell even when oppositely charged.

Enhanced conductivity of thin film polyaniline by self-assembled transition metal complexes.

In a recent study, the transition metal complex, cis-dichlorobis(2-,2'-dipyridyl)ruthenium (II) (Ru(bpy)2Cl2), and the macrocycle Ru(TPP)CO (TPP:- tetraphenylporphine) were bound to pyridine terminated self-assembled monolayers on quartz. Following modification of the quartz surface with metal complexes, the conducting polymer polyaniline was deposited via in situ polymerization. The sheet conductivity (as measured by the four-probe method) of the resulting polyaniline films deposited onto Ru(bpy)2Cl2 and Ru(TPP)CO surfaces was significantly enhanced relative to films deposited onto unmodified quartz. It is postulated that either the macrocycle or the transition metal complex-modified surface interacts with the conducting polymer as it is forming, resulting in a more ordered expanded coil conformation for the polymer. The net result of such an interaction is a thin film possessing significantly greater electrical conductivity.