A non-invasive, sensor-based approach to exploit the autofluorescence of saffron (Crocus sativus L.) for on-site evaluation of aging.

So far, compliance with ISO 3632 standard specifications for top-quality saffron guarantees good agricultural and post-harvest production practices. Tracking early-stage oxidation remains challenging. Our study aims to address this issue by exploring the visible, fluorescence, and near-infrared spectra of category I saffron. Using a multi-spectral sensor, we tested fresh and artificially aged saffron in powder form. High autofluorescence intensities at 600-700 nm allowed calibration for the 'content of aged saffron'. Samples with minimum coloring strength (200-220 units) were classified as 70% aged, while those exceeding maximum aroma strength (50 units) as 100% aged. Consistent patterns across origin, age, and processing history indicated potential for objectively assessing early-oxidation markers. Further analyses uncovered multiple contributing fluorophores, including cis-apocarotenoids, correlated with FTIR-based aging markers. Our findings underscore that sensing autofluorescence of traded saffron presents an innovative quality diagnostic approach, paving new research pathways for assessing the remaining shelf-life along its supply chain.

High conductivity of ultrathin nanoribbons of SrRuO3 on SrTiO3 probed by infrared spectroscopy.

SrRuO3 (SRO) is a perovskite increasingly used in oxide-based electronics both for its intrinsic metallicity, which remains unaltered in thin films and for the ease of deposition on dielectric perovskites like SrTiO3, (STO) to implement SRO/STO microcapacitors and other devices. In order to test the reliability of SRO/STO also as high-current on-chip conductor, when the SRO dimensions are pushed to the nanoscale, here we have measured the electrodynamic properties of arrays of nanoribbons, fabricated by lithography starting from an ultrathin film of SRO deposited on a STO substrate. The nanoribbons are 6 or 4 nm thick, 400, 200 and 100 nm wide and 5 mm long. The measurements have been performed by infrared spectroscopy, a non-contact weakly perturbing technique which also allows one to separately determine the carrier density and their scattering rate or mobility. Far-infrared reflectivity spectra have been analyzed by Rigorous Coupled-Wave Analysis (RCWA) and by an Effective Medium Theory, obtaining consistent results. With the radiation polarized along the nanoribbons, we obtain a carrier density similar to that of a flat film used as reference, which in turn is similar to that of bulk SRO. Moreover, in the nanoribbons the carrier scattering rate is even smaller than in the unpatterned film by about a factor of 2. This shows that the transport properties of SRO deposited on STO remain at least unaltered down to nanometric dimensions, with interesting perspectives for implementing on-chip nano-interconnects in an oxide-based electronics. When excited in the perpendicular direction, the nanoribbons appear instead virtually transparent to the radiation field, as predicted by RCWA.

Human insulin fibrillogenesis in the presence of epigallocatechin gallate and melatonin: Structural insights from a biophysical approach.

Fibrillogenesis of monomeric human insulin in the presence or absence of (-)-epigallocatechin-3-gallate and melatonin was here investigated using a multi-technique approach. Results from Raman and Infrared spectroscopy pointed out that a high content of intermolecular ß-sheet aggregates is formed after long-term incubation. However, near UV experiments, Dynamic Light Scattering, Thioflavin-T fluorescence measurements and Atomic Force Microscopy revealed that the kinetics from native-to-fibrillar state of insulin is hampered only in the presence of (-)-epigallocatechin-3-gallate. Molecular dynamic simulations indicated that this compound binds near the B11-B18 protein segment, where hydrophobic residues responsible for the beginning of cooperative aggregation are located. Such a preferential binding region is not recognized by melatonin, a highly mobile molecule, which indeed does not affect fibril formation. The results of the present study demonstrate that (-)-epigallocatechin-3-gallate interferes with the insulin nucleation phase, giving rise to amorphous aggregates in the early stages of the aggregation process.

Protein clustering in chemically stressed HeLa cells studied by infrared nanospectroscopy.

Photo-Thermal Induced Resonance (PTIR) nanospectroscopy, tuned towards amide-I absorption, was used to study the distribution of proteic material in 34 different HeLa cells, of which 18 were chemically stressed by oxidative stress with Na3AsO3. The cell nucleus was found to provide a weaker amide-I signal than the surrounding cytoplasm, while the strongest PTIR signal comes from the perinuclear region. AFM topography shows that the cells exposed to oxidative stress undergo a volume reduction with respect to the control cells, through an accumulation of the proteic material around and above the nucleus. This is confirmed by the PTIR maps of the cytoplasm, where the pixels providing a high amide-I signal were identified with a space resolution of ∼300 × 300 nm. By analyzing their distribution with two different statistical procedures we found that the probability to find protein clusters smaller than 0.6 µm in the cytoplasm of stressed HeLa cells is higher by 35% than in the control cells. These results indicate that it is possible to study proteic clustering within single cells by label-free optical nanospectroscopy.

Role of dietary antioxidant (-)-epicatechin in the development of ß-lactoglobulin fibrils.

Under specific physico-chemical conditions ß-lactoglobulin is seen to form fibrils structurally highly similar to those that are formed by the amyloid-like proteins associated with neurodegenerative disorders, such as Alzheimer and Parkinson diseases. In the present study we provide insights on the possible role that the dietary flavonoid (-)-epicatechin plays on ß-lactoglobulin fibril formation. Fibril formation is induced by keeping ß-lactoglobulin solutions at pH2.0 and at a temperature of 80°C for 24h. Atomic Force Microscopy measurements suggest that, by adding (-)-epicatechin in the solution, fibrils density is visibly lowered. This last observation is confirmed by Fluorescence Correlation Spectroscopy experiments. With the use of Fourier Transform IR spectroscopy we monitored the relative abundances of the secondary structures components during the heating process. We observed that in the presence of (-)-epicatechin the spectral-weight exchange between different secondary structures is partially inhibited. Molecular Dynamics simulations have been able to provide an atomistic explanation of this experimental observation, showing that (-)-epicatechin interacts with ß-lactoglobulin mainly via the residues that, normally in the absence of (-)-epicatechin, are involved in ß-sheet formation. Unveiling this molecular mechanism is an important step in the process of identifying suitable molecules apt at finely tuning fibril formation like it is desirable to do in food industry applications.

Mapping the amide I absorption in single bacteria and mammalian cells with resonant infrared nanospectroscopy.

Infrared (IR) nanospectroscopy performed in conjunction with atomic force microscopy (AFM) is a novel, label-free spectroscopic technique that meets the increasing request for nano-imaging tools with chemical specificity in the field of life sciences. In the novel resonant version of AFM-IR, a mid-IR wavelength-tunable quantum cascade laser illuminates the sample below an AFM tip working in contact mode, and the repetition rate of the mid-IR pulses matches the cantilever mechanical resonance frequency. The AFM-IR signal is the amplitude of the cantilever oscillations driven by the thermal expansion of the sample after absorption of mid-IR radiation. Using purposely nanofabricated polymer samples, here we demonstrate that the AFM-IR signal increases linearly with the sample thickness t for t > 50 nm, as expected from the thermal expansion model of the sample volume below the AFM tip. We then show the capability of the apparatus to derive information on the protein distribution in single cells through mapping of the AFM-IR signal related to the amide-I mid-IR absorption band at 1660 cm(-1). In Escherichia Coli bacteria we see how the topography changes, observed when the cell hosts a protein over-expression plasmid, are correlated with the amide I signal intensity. In human HeLa cells we obtain evidence that the protein distribution in the cytoplasm and in the nucleus is uneven, with a lateral resolution better than 100 nm.

Intrinsic linewidth of the plasmonic resonance in a micrometric metal mesh.

The intrinsic linewidth and angular dispersion of Surface Plasmon Polariton resonance of a micrometric metal mesh have been measured with a collimated mid-infrared beam, provided by an External Cavity tunable Quantum Cascade Laser. We show that the use of a collimated beam yields an observed resonance linewidth γ = 12 cm(-1) at the resonance frequency ν0 = 1658 cm(-1), better by an order of magnitude than with a non-collimated beam. The extremely narrow plasmon resonance attained by our mesh is then exploited to reconstruct, by varying the QCL angle of incidence θ, the angular intensity distribution f(θ) of a globar at the focal plane of a conventional imaging setup. We thus show that f(θ) is better reproduced by a Gaussian distribution than by a uniform one, in agreement with ray-tracing simulation.

Relationship between digestibility and secondary structure of raw and thermally treated legume proteins: a Fourier transform infrared (FT-IR) spectroscopic study.

The secondary structure of proteins in legumes, cereals, milk products and chicken meat was studied by diffuse reflectance infrared spectroscopy in the region of the amide I band. Major secondary structure components ( ß-sheets, random coil, α-helix, turns), together with the low- and high-frequency side contributions, were resolved and related to the in vitro digestibility behaviour of the different foods. A strong inverse correlation between the relative spectral weights of the ß-sheet structures and in vitro protein digestibility values was measured. Structural modifications in legume proteins induced by autoclaving were monitored by the changes in the amide I spectra. The results indicate that the ß-sheet structures of raw legume proteins and the intermolecular ß-sheet aggregates, arising upon heating, are primary factors in adversely affecting the digestibility.

Secondary structure of food proteins by Fourier transform spectroscopy in the mid-infrared region.

Fourier transform spectroscopy in the mid-infrared (400-5,000 cm(-1)) (FT-IR) is being recognized as a powerful tool for analyzing chemical composition of food, with special concern to molecular architecture of food proteins. Unlike other spectroscopic techniques, it provides high-quality spectra with very small amount of protein, in various environments irrespective of the molecular mass. The fraction of peptide bonds in alpha-helical, beta-pleated sheet, turns and aperiodic conformations can be accurately estimated by analysis of the amide I band (1,600-1,700 cm(-1)) in the mid-IR region. In addition, FT-IR measurement of secondary structure highlights the mechanism of protein aggregation and stability, making this technique of strategic importance in the food proteomic field. Examples of applications of FT-IR spectroscopy in the study of structural features of food proteins critical of nutritional and technological performance are discussed.

Observation of charge-density-wave excitations in manganites.

In the optical conductivity of four different manganites with commensurate charge order (CO), strong peaks appear in the meV range below the ordering temperature T_{CO}. They are similar to those reported for one-dimensional charge density waves (CDW) and are assigned to pinned phasons. The peaks and their overtones allow one to obtain, for La_{1-n/8}Ca_{n/8}MnO_{3} with n=5, 6, the electron-phonon coupling, the effective mass of the CO system, and its contribution to the dielectric constant. These results support a description of the CO in La-Ca manganites in terms of moderately weak coupling and of the CDW theory.

Far-infrared absorption of La1-xCaxMnO3-y at high pressure.

The first far-infrared absorption spectra of manganite samples at pressures P up to 10 GPa were obtained on La1-xCaxMnO3-y by use of synchrotron radiation. For x=0.25 and 0.20 (y=0), P promotes partial metallization at room temperature through a strong reduction of the insulating gap. An x=0.20 sample with y=0.08 does not show any charge delocalization effect up to 10 GPa. An Urbach-like model of disordered Jahn-Teller wells is shown to well fit the far-infrared band edge and allows one to obtain a reliable pressure dependence of the energy gap.

A comparative EPR, infrared and Raman study of natural and deproteinated tooth enamel and dentin.

The aim of the present work was to investigate how the native signal observed in the electron paramagnetic resonance (EPR) spectrum of tooth enamel and dentin is associated with the organic content of the two tissues. This was achieved by comparing the EPR native signal and the optical bands (Raman and infrared, IR) associated with organic components of tooth enamel and dentin, in natural and deproteinated samples. The main results were: (a) in natural samples, the organic optical bands are more intense in dentin than in enamel, in contrast with the EPR native signal which shows similar intensity in the two tissues; (b) after deproteination, the optical organic bands are completely suppressed in both dentin and enamel, while the EPR native signal is eliminated only in dentin. It is suggested that the IR and Raman organic bands are originated in the bulk of the organic matrix, while the paramagnetic centres associated with the EPR native signal are located in the organic-mineral interface.

Spectral distribution of infrared synchrotron radiation by an insertion device and its edges: A comparison between experimental and simulated spectra

The first measurements of the spectral distribution of infrared radiation emitted by an undulator are reported. They are compared with calculations including both velocity and acceleration terms. Measurements have been performed at the beam line SIRLOIN (Spectroscopie en InfraRouge Lointain). The agreement between the observations and this first exact numerical solution shows that the inclusion of the velocity term in the submillimeter frequency range is necessary. Moreover, structures due to undulator edges are observed in the far infrared and mid-infrared range, while the interference pattern due to redshifted harmonics of the undulator is dominating in the mid-infrared to near infrared.

Infrared properties of chemical-vapor deposition polycrystalline diamond windows.

Low-resolution transmittance and reflectance spectra of high-quality chemical-vapor deposition (CVD) diamond windows were measured in the infrared in the 2.5-500-mum wavelength range (20-4000 cm(-1)). High-resolution measurements on a window with nearly parallel surfaces show well defined interference fringes at low frequencies. By standard procedures the optical constants n and k of CVD diamond were determined, for the first time to the author's knowledge, in the far-infrared region. It is shown that a window with a large wedge angle, close to 1 degrees , does not produce appreciable interference fringes. Modeling of these results confirms that interference fringes can be avoided by use of properly wedged CVD diamond windows. This result is of considerable relevance to the use of CVD diamond windows in spectroscopic applications for which fringe suppression is a major requirement.

SINBAD, a brilliant IR source from the DAPhiNE storage ring.

SINBAD is the Italian IR synchrotron radiation beamline, designed to work at wavelengths greater than 10 micro m. It is being installed on DAPhiNE, a new collider that is designed to work at 0.51 GeV with a beam current up to 5 A. Due to such a high current, the IR extracted from a bending magnet will be more brilliant than that of a black body at 2000 K by two orders of magnitude at 100 micro m. The beamline optical system, projected by ray-tracing simulation, consists of six mirrors that first focus the radiation on a wedged CVD diamond-film window and then transfer the collimated beam to the experimental area where a Michelson interferometer will be installed.