A rationally designed synthetic antimicrobial peptide against Pseudomonas-associated corneal keratitis: Structure-function correlation.

Contact lens wearers are at an increased risk of developing Pseudomonas-associated corneal keratitis, which can lead to a host of serious ocular complications. Despite the use of topical antibiotics, ocular infections remain a major clinical problem, and a strategy to avoid Pseudomonas-associated microbial keratitis is urgently required. The hybrid peptide VR18 (VARGWGRKCPLFGKNKSR) was designed to have enhanced antimicrobial properties in the fight against Pseudomonas-induced microbial keratitis, including contact lens-related keratitis. In this paper, VR18's modes of action against Pseudomonas membranes were shown by live cell Raman spectroscopy, live cell NMR, live-cell fluorescence microscopy and measures taken using sparsely tethered bilayer lipid membrane bacterial models to be via a bacterial-specific membrane disruption mechanism. The high affinity and selectivity of the peptide were then demonstrated using in vivo, in vitro and ex vivo models of Pseudomonas infection. The extensive data presented in this work suggests that topical employment of the VR18 peptide would be a potent therapeutic agent for the prevention or remedy of Pseudomonas-associated microbial keratitis.

Lattice dynamics of Ge1-xSnx alloy nanowires.

Alloying group IV semiconductors offers an effective way to engineer their electronic properties and lattice dynamics. The incorporation of Sn in Ge permits a transition from an indirect to a direct bandgap semiconductor. Here, by combining polarization, laser power-dependent and temperature-dependent micro-Raman spectroscopy we explore the full lattice dynamics of Ge1-xSnx (x = 0.01, 0.06 and 0.08) alloy nanowires. In the high Sn content samples (x ≥ 0.06), a low-frequency tail and a high-frequency shoulder are observed which are associated with the F2g optical phonon mode of Ge (Ge-Ge mode). The new modes are assigned to the stretching of Ge-Ge bonds due to Sn-induced lattice relaxation and compression, respectively. The symmetry of the observed Raman modes has been studied by polarization-dependent Raman scattering. Nonlinear fitting of the laser power-dependent intensity of the high-frequency Ge-Ge mode in the Ge1-xSnx alloy nanowires with x = 0.06 and 0.08 suggests the activation of a third-order stimulated Raman scattering process, due to the high intensity localized electric field surrounding the Sn clusters. Finally, from the temperature-dependent Raman study, we have estimated the isobaric Grüneisen parameters for all the observed modes.

One-Step Grown Carbonaceous Germanium Nanowires and Their Application as Highly Efficient Lithium-Ion Battery Anodes.

Developing a simple, cheap, and scalable synthetic method for the fabrication of functional nanomaterials is crucial. Carbon-based nanowire nanocomposites could play a key role in integrating group IV semiconducting nanomaterials as anodes into Li-ion batteries. Here, we report a very simple, one-pot solvothermal-like growth of carbonaceous germanium (C-Ge) nanowires in a supercritical solvent. C-Ge nanowires are grown just by heating (380-490 °C) a commercially sourced Ge precursor, diphenylgermane (DPG), in supercritical toluene, without any external catalysts or surfactants. The self-seeded nanowires are highly crystalline and very thin, with an average diameter between 11 and 19 nm. The amorphous carbonaceous layer coating on Ge nanowires is formed from the polymerization and condensation of light carbon compounds generated from the decomposition of DPG during the growth process. These carbonaceous Ge nanowires demonstrate impressive electrochemical performance as an anode material for Li-ion batteries with high specific charge values (>1200 mAh g-1 after 500 cycles), greater than most of the previously reported for other "binder-free" Ge nanowire anode materials, and exceptionally stable capacity retention. The high specific charge values and impressively stable capacity are due to the unique morphology and composition of the nanowires.

Growth and analysis of the tetragonal (ST12) germanium nanowires.

New semiconducting materials, such as state-of-the-art alloys, engineered composites and allotropes of well-established materials can demonstrate unique physical properties and generate wide possibilities for a vast range of applications. Here we demonstrate, for the first time, the fabrication of a metastable allotrope of Ge, tetragonal germanium (ST12-Ge), in nanowire form. Nanowires were grown in a solvothermal-like single-pot method using supercritical toluene as a solvent, at moderate temperatures (290-330 °C) and a pressure of ∼48 bar. One-dimensional (1D) nanostructures of ST12-Ge were achieved via a self-seeded vapour-liquid-solid (VLS)-like paradigm, with the aid of an in situ formed amorphous carbonaceous layer. The ST12 phase of Ge nanowires is governed by the formation of this carbonaceous structure on the surface of the nanowires and the creation of Ge-C bonds. The crystalline phase and structure of the ST12-Ge nanowires were confirmed by X-ray diffraction (XRD), high-resolution transmission electron microscopy (HRTEM) and Raman spectroscopy. The nanowires produced displayed a high aspect ratio, with a very narrow mean diameter of 9.0 ± 1.4 nm, and lengths beyond 4 µm. The ST12-Ge nanowire allotrope was found to have a profound effect on the intensity of the light emission and the directness of the bandgap, as confirmed by a temperature-dependent photoluminescence study.

Solution phase growth and analysis of super-thin zigzag tin selenide nanoribbons.

Tin selenide (SnSe), a highly promising layered material, has been garnering particular interest in recent times due to its significant promise for future energy devices. Herein we report a simple solution-phase approach for growing highly crystalline layered SnSe nanoribbons. Polyvinylpyrrolidone (PVP) was used as a templating agent to selectively passivates the (100) and (001) facets of the SnSe nanoribbons resulting in the unique growth of nanoribbons along theirb-axis with a defined zigzag edge state along the sidewalls. The SnSe nanoribbons are few layers thick (∼20 layers), with mean widths of ∼40 nm, and achievable length of >1µm. Nanoribbons could be produced in relatively high quantities (>150 mg) in a single batch experiment. The PVP coating also offers some resistance to oxidation, with the removal of the PVP seen to lead to the formation of a SnSe/SnOxcore-shell structure. The use of non-toxic PVP to replace toxic amines that are typically employed for other 1D forms of SnSe is a significant advantage for sustainable and environmentally friendly applications. Heat transport properties of the SnSe nanoribbons, derived from power-dependent Raman spectroscopy, demonstrate the potential of SnSe nanoribbons as thermoelectric material.

Probing dipole and quadrupole resonance mode in non-plasmonic nanowire using Raman spectroscopy.

Electric field enhancement in semiconductor nanostructures offers a possibility to find an alternative to the metallic particles which is well known for tuning the light-matter interaction due to its strong polarizability and size-dependent surface plasmon resonance energy. Raman spectroscopy is a powerful technique to monitor the electric field as its scattering depends on the electromagnetic eigenmode of the particle. Here, we observe enhanced polarized Raman scattering from germanium nanowires of different diameters. The incident electromagnetic radiation creates a distribution of the internal electric field inside the naowires which can be enhanced by manipulating the nanowire diameter, the incident electric field and its polarization. Our estimation of the enhancement factor, including its dependence on nanowire diameter, agrees well with the Mie theory for an infinite cylinder. Furthermore, depending on diameter of nanowire and wavelength of incident radiation, polarized Raman study shows dipolar (antenna effect) and quadrupolar resonances, which has never been observed in germanium nanowire. We attempt to understand this polarized Raman behavior using COMSOL Multiphysics simulation, which suggests that the pattern observed is due to photon confinement within the nanowires. Thus, the light scattering direction can be toggled by tuning the polarization of incident excitation and diameter of non plasmonic nanowire.

Targeted inhibition of amyloidogenesis using a non-toxic, serum stable strategically designed cyclic peptide with therapeutic implications.

Amyloidogenic disorders are currently rising as a global health issue, prompting more and more studies dedicated to the development of effective targeted therapeutics. The innate affinity of these amyloidogenic proteins towards the biomembranes adds further complexities to the systems. Our previous studies have shown that biologically active peptides can effectively target amyloidogenesis serving as an efficient therapeutic alternative in several amyloidogenic disorders. The structural uniqueness of the PWWP motif in the de novo designed heptapeptide, KR7 (KPWWPRR-NH2) was demonstrated to target insulin fiber elongation specifically. By working on insulin, an important model system in amyloidogenic studies, we gained several mechanistic insights into the inhibitory actions at the protein-peptide interface. Here, we report a second-generation non-toxic and serum stable cyclic peptide, based primarily on the PWWP motif that resulted in complete inhibition of insulin fibrillation both in the presence and absence of the model membranes. Using both low- and high-resolution spectroscopic techniques, we could delineate the specific mechanism of inhibition, at atomistic resolution. Our studies put forward an effective therapeutic intervention that redirects the default aggregation kinetics towards off-pathway fibrillation. Based on the promising results, this novel cyclic peptide can be considered an excellent lead to design pharmaceutical molecules against amyloidogenesis.

Size-dependent apoptotic activity of gold nanoparticles on osteosarcoma cells correlated with SERS signal.

In the last decade, gold nanoparticles have emerged as promising agents for in vitro bio-sensing and in vivo cancer theranostics. However, different investigations have reported widely varying cytotoxicity and uptake efficiency of gold nanoparticles depending upon their size. Therefore, more extensive studies are needed to standardize these biological effects as a function of size on a particular cell line. In addition, to obtain robust confirmation on the correlation of a size to biological effect, thorough mechanistic study must also be performed. In this study, the size dependent biological activities of gold nanoparticles on osteosarcoma cells is investigated towards exploring their potential theranostic application in bone cancer, for which very scarce literature reports are available. Tris-assisted citrate based method was optimized to synthesize stable gold naoparticles of 40-60 nm sizes. Nanoparticles were characterized through UV-Vis spectroscopy, field emission scanning electron microscope (FESEM) and dynamic light scattering (DLS). Increasing concentrations of gold nanoparticles (AuNPs) of 46 nm size, enhanced the rate of reactive oxygen species (ROS)-induced apoptosis in MG63 cells by disrupting their mitochondrial membrane potential. Considerably higher cell death was observed for 46 and 60 nm AuNPs compared to 38 nm at all concentrations of 200, 400 and 800 ng/mL. Further, molecular signatures of cellular apoptosis under nanoparticle treatment were optically assessed through surface enhanced Raman scattering (SERS). A significant Raman enhancement in cancer cells under treatment of larger gold nanoparticles (46 and 60 nm) at fixed wavelength of 785 nm and laser power of 8.0 mW was evident. In corroboration with molecular biology techniques, SERS observation confirmed the size-dependent apoptotic phenomena in osteosarcoma cells under treatment of gold nanoparticles. Study demonstrates a facile, non-active targeting approach for detection of size-dependent AuNP-induced apoptosis in osteosarcoma cells through label-free SERS method.

Molecular Details of a Salt Bridge and Its Role in Insulin Fibrillation by NMR and Raman Spectroscopic Analysis.

Insulin, a simple polypeptide hormone with huge biological importance, has long been known to self-assemble in vitro and form amyloid-like fibrillar aggregates. Utilizing high-resolution NMR, Raman spectroscopy, and computational analysis, we demonstrate that the fluctuation of the carboxyl terminal (C-ter) residues of the insulin B-chain plays a key role in the growth phase of insulin aggregation. By comparing the insulin sourced from bovine, human, and the modified glargine (GI), we observed reduced aggregation propensity in the GI variant, resulting from two additional Arg residues at its C-ter. NMR analysis showed atomic contacts and residue-specific interactions, particularly the salt bridge and H-bond formed among the C-ter residues Arg31B, Lys29B, and Glu4A. These inter-residue interactions were reflected in strong nuclear Overhauser effects among Arg31BδH-Glu4AδH and Lys29BδHs-Glu4AδH in GI, as well as the associated downfield chemical shift of several A-chain amino terminal (N-ter) residues. The two additional Arg residues of GI, Arg31B and Arg32B, enhanced the stability of the GI native structure by strengthening the Arg31B, Lys29B, and Glu4A salt bridge, thus reducing extensive thermal distortion and fluctuation of the terminal residues. The high stability of the salt bridge retards tertiary collapse, a crucial biochemical event for oligomerization and subsequent fibril formation. Circular dichroism and Raman spectroscopic measurement also suggest slow structural distortion in the early phase of the aggregation of GI because of the restricted mobility of the C-ter residues as explained by NMR. In addition, the structural and dynamic parameters derived from molecular dynamics simulations of insulin variants highlight the role of residue-specific contacts in aggregation and amyloid-like fibril formation.

Sequence specificity of amylin-insulin interaction: a fragment-based insulin fibrillation inhibition study.

Subcutaneous insulin delivery serves as the major treatment for the ever-increasing spread of type II diabetes worldwide. However, long-term exposure to insulin results in local aggregates at the site of injection. This therapeutic concern accentuates the need to develop newer effective excipients to stabilize the insulin in pharmaceutical formulations. The fact that in normal physiological conditions, insulin interacts with the amylin hormone co-secreted from the pancreas, we targeted a peptide-mimetic approach based on the amylin sequence. The amylin-fibrillating core (NL6- N22FGAIL27 from the human Islet Amyloid Poly-Peptide) and its derivative NFGAXL (NL6X, X = 2-aminobenzoic acid) were used as potential inhibitory peptides against insulin amyloidogenesis. The fibrillation kinetics in the presence of the inhibitors was studied using an array of biophysical and microscopic techniques. High-resolution NMR spectroscopy enabled probing of the inhibitory interaction at an atomic resolution. Our results highlight the potential of using the naturally evolved NL6 peptide as an effective inhibitor against insulin fibrillation.

Chemometric analysis of integrated FTIR and Raman spectra obtained by non-invasive exfoliative cytology for the screening of oral cancer.

FTIR spectroscopy and Raman spectroscopy of biological analytes are increasingly explored as screening tools for early detection of cancer. In the present study, an integrated analysis of the FTIR and Raman spectra obtained from exfoliated cells is adopted to improve discrimination of normal, pre-cancerous and cancerous conditions. Multiple spectra were obtained from 13 normal, 13 pre-cancer and 10 cancer patients in both modes. Compared to normal patients, significant differences were observed at 1550, 1580, 1640, 2370, 2330, 2950-3000 and 3650-3750 cm-1 (FTIR) and 520, 640, 785, 827, 850, 935, 1003, 1175, 1311 cm-1 and 1606 cm-1 (Raman) vibrations of the other two. The increase in DNA, protein and lipid content with malignancy was more clearly elucidated by examining both spectra. Principal component analysis (PCA)-linear discriminant analysis (LDA) with 10-fold cross validation of the FTIR and Raman spectral data sets showed efficient discrimination between normal and pathological conditions while overlapping was seen between the two pathologies. The PCA-LDA model of the dual spectra yielded a classification accuracy of 98% in comparison with either FTIR (85%) or Raman (82%) in a spectrum-wise comparison. In the patient-wise approach (mean of all spectra from a patient), the overall classification efficiency was 73%, 80% and 87% for FTIR, Raman and integrated spectral approaches respectively. Moreover, the efficiency of the integrated FTIR-Raman PCA-LDA model as a prediction tool was tested to screen susceptible individuals (11 cigarette smokers) using the dual spectra acquired from these individuals. The study presents proof-of-concept for adopting a large-scale, follow-up trial of the approach for mass screening purposes.