Raman Spectroscopic Analysis of Steviol Glycosides: Spectral Database and Quality Control Algorithms.

Besides all sharing an extraordinary high (i.e., up to ~450 times) sweetening power as compared to sucrose and while presenting strong similarities in their molecular structures, molecules belonging to the family of diterpene glycosides (i.e., the secondary metabolites of Stevia rebaudiana) differ in specific structural details that strongly impact on their levels of sweetness and bitter aftertaste. Given the nutritional and pharmacological benefits of steviol secondary metabolites as natural dietetic and anti-diabetic remedies, extraction and purification of steviol glycosides from plant material are nowadays widely spread among many countries. However, an unpleasant bitter aftertaste, which is linked to a genetic variation in human bitter taste receptors, hampers the full exploitation of such benefits and calls for a prompt improvement in organoleptic property control of stevia products. A deeper understanding of the molecular structure of different steviol glycosides and the consequent development of promptly measurable criteria for the organoleptic performance of their mixtures will support processing optimization and control of taste profiles within desired yields. The present research aimed at establishing Raman spectroscopic algorithms for quantitative characterizations of raw stevia-based sweetener products. First, a series of twelve high-purity diterpene glycosides were analyzed by high spectrally resolved Raman spectroscopy and their spectra analyzed in order to establish a complete Raman library of molecular structures. Then, quantitative spectroscopic parameters were built up and applied to characterize the organoleptic property of five different commercially available samples including the recently developed Rebaudioside M isoform. Raman spectroscopy was confirmed as a versatile analytical technique that could be used for quantitative quality control tasks on the production line and for prompt in situ characterizations of purchased products.

Effect of rapid cooling on residual stress and surface fracture toughness of dental zirconia.

Short-time sintering of dental zirconia not only improves manufacturing efficiency of zirconia prosthetics, but also enables an attractive situation in which prosthetic treatment can be completed within a single visit. Although many studies have clarified the effects of heating rate and dwell time on the properties of dental zirconia during short-time sintering, there are only a few studies on rapid cooling. In this study, we investigated the effect of cooling rate on dental zirconia. It was found that the cooling rate had no effect on the three-point flexural strength, but a fast cooling rate improved fracture toughness at the material surface. Raman piezo-spectroscopy showed that a compressive stress layer formed in the neighborhood of the zirconia surface and that its thickness increased with increasing cooling rate. From the above results, it was concluded that the compressive stress layer formed on the surface by rapid cooling improved the apparent fracture toughness at the material surface.

Temperature-Dependent Structural Properties of Nickel and Cobalt Selenite Hydrates as Solar Water Evaporators.

Solar water evaporation offers a promising solution to address global water scarcity, utilizing renewable energy for purification and desalination. Transition-metal selenite hydrates (specifically nickel and cobalt) have shown potential as solar absorbers with high evaporation rates of 1.83 and 2.34 kg∙m-2∙h-1, but the reported discrepancy in evaporation rate deserves further investigation. This investigation aims to clarify their thermal stability for applications and determine the underlying mechanisms responsible for the differences. Nickel and cobalt selenite hydrate compositions were synthesized and investigated via thermogravimetric analysis, X-ray diffraction, and Raman spectroscopy to assess their temperature-induced structural and compositional variations. The results reveal distinct phase transitions and structural alterations under various temperature conditions for these two photothermal materials, providing valuable insights into the factors influencing water transportation and evaporation rates.

Exploring the cellular antioxidant mechanism against cytotoxic silver nanoparticles: a Raman spectroscopic analysis.

Silver nanoparticles (AgNPs) hold great promise for several different applications, from colorimetric sensors to antimicrobial agents. Despite their widespread incorporation in consumer products, limited understanding of the detrimental effects and cellular antioxidant responses associated with AgNPs at sublethal concentrations persists, raising concerns for human and ecological well-being. To address this gap, we synthesized AgNPs of varying sizes and evaluated their cytotoxicity against human dermal fibroblasts (HDF). Our study revealed that toxicity of AgNPs is a time- and size-dependent process, even at low exposure levels. AgNPs exhibited low short-term cytotoxicity but high long-term impact, particularly for the smallest NPs tested. Raman microspectroscopy was employed for in-time investigations of intracellular molecular variations during the first 24 h of exposure to AgNPs of 35 nm. Subtle protein and lipid degradations were detected, but no discernible damage to the DNA was observed. Signals associated with antioxidant proteins, such as superoxide dismutase (SOD), catalase (CAT) and metallothioneins (MTs), increased over time, reflecting the heightened production of these defense agents. Fluorescence microscopy further confirmed the efficacy of overexpressed antioxidant proteins in mitigating ROS formation during short-term exposure to AgNPs. This work provides valuable insights into the molecular changes and remedial strategies within the cellular environment, utilizing Raman microspectroscopy as an advanced analytical technique. These findings offer a novel perspective on the cytotoxicity mechanism of AgNPs, contributing to the development of safer materials and advice on regulatory guidelines for their biomedical applications.

Ammonia Toxicity and Associated Protein Oxidation: A Single-Cell Surface Enhanced Raman Spectroscopy Study.

Ammonia (NH3) is a commonly used industrial chemical to which exposure at high concentrations can result in severe skin damage. Moreover, high levels of ammonia in the human body can lead to hyperammonemia conditions and enhanced cancer metabolism. In this work, the toxicity mechanism of NH3 has been studied against human dermal fibroblast (HDF) cells using surface-enhanced Raman spectroscopy (SERS). For this purpose, gold nanoparticles of size 50 nm have been prepared and used as probes for Raman signal enhancement, after being internalized inside HDF cells. Following the exposure to ammonia, HDF cells showed a significant variation in the protein ternary structure's signals, demonstrating their denaturation and oxidation process, together with early signs of apoptosis. Meaningful changes were observed especially in the Raman vibrations of sulfur-containing amino acids (cysteine and methionine) together with aromatic residues. Fluorescence microscopy revealed the formation of reactive oxygen and nitrogen species in cells, which confirmed their stressed condition and to whom the causes of protein degradation can be attributed. These findings can provide new insights into the mechanism of ammonia toxicity and protein oxidation at a single-cell level, demonstrating the high potential of the SERS technique in investigating the cellular response to toxic compounds.

Raman Multi-Omic Snapshot and Statistical Validation of Structural Differences between Herpes Simplex Type I and Epstein-Barr Viruses.

Raman spectroscopy was applied to study the structural differences between herpes simplex virus Type I (HSV-1) and Epstein-Barr virus (EBV). Raman spectra were first collected with statistical validity on clusters of the respective virions and analyzed according to principal component analysis (PCA). Then, average spectra were computed and a machine-learning approach applied to deconvolute them into sub-band components in order to perform comparative analyses. The Raman results revealed marked structural differences between the two viral strains, which could mainly be traced back to the massive presence of carbohydrates in the glycoproteins of EBV virions. Clear differences could also be recorded for selected tyrosine and tryptophan Raman bands sensitive to pH at the virion/environment interface. According to the observed spectral differences, Raman signatures of known biomolecules were interpreted to link structural differences with the viral functions of the two strains. The present study confirms the unique ability of Raman spectroscopy for answering structural questions at the molecular level in virology and, despite the structural complexity of viral structures, its capacity to readily and reliably differentiate between different virus types and strains.

Raman Fingerprints of SARS-CoV-2 Omicron Subvariants: Molecular Roots of Virological Characteristics and Evolutionary Directions.

The latest RNA genomic mutation of SARS-CoV-2 virus, termed the Omicron variant, has generated a stream of highly contagious and antibody-resistant strains, which in turn led to classifying Omicron as a variant of concern. We systematically collected Raman spectra from six Omicron subvariants available in Japan (i.e., BA.1.18, BA.2, BA.4, BA.5, XE, and BA.2.75) and applied machine-learning algorithms to decrypt their structural characteristics at the molecular scale. Unique Raman fingerprints of sulfur-containing amino acid rotamers, RNA purines and pyrimidines, tyrosine phenol ring configurations, and secondary protein structures clearly differentiated the six Omicron subvariants. These spectral characteristics, which were linked to infectiousness, transmissibility, and propensity for immune evasion, revealed evolutionary motifs to be compared with the outputs of genomic studies. The availability of a Raman "metabolomic snapshot", which was then translated into a barcode to enable a prompt subvariant identification, opened the way to rationalize in real-time SARS-CoV-2 activity and variability. As a proof of concept, we applied the Raman barcode procedure to a nasal swab sample retrieved from a SARS-CoV-2 patient and identified its Omicron subvariant by coupling a commercially available magnetic bead technology with our newly developed Raman analyses.

Raman Multi-Omic Snapshots of Koshihikari Rice Kernels Reveal Important Molecular Diversities with Potential Benefits in Healthcare.

This study exploits quantitative algorithms of Raman spectroscopy to assess, at the molecular scale, the nutritional quality of individual kernels of the Japanese short-grain rice cultivar Koshihikari in terms of amylose-to-amylopectin ratio, fractions of phenylalanine and tryptophan aromatic amino acid residues, protein-to-carbohydrate ratio, and fractions of protein secondary structures. Statistical assessments on a large number of rice kernels reveal wide distributions of the above nutritional parameters over nominally homogeneous kernel batches. This demonstrates that genetic classifications cannot catch omic fluctuations, which are strongly influenced by a number of extrinsic factors, including the location of individual grass plants within the same rice field and the level of kernel maturation. The possibility of collecting nearly real-time Raman "multi-omic snapshots" of individual rice kernels allows for the automatic (low-cost) differentiation of groups of kernels with restricted nutritional characteristics that could be used in the formulation of functional foods for specific diseases and in positively modulating the intestinal microbiota for protection against bacterial infection and cancer prevention.

In Situ Raman Study of Neurodegenerated Human Neuroblastoma Cells Exposed to Outer-Membrane Vesicles Isolated from Porphyromonas gingivalis.

The aim of this study was to elucidate the chemistry of cellular degeneration in human neuroblastoma cells upon exposure to outer-membrane vesicles (OMVs) produced by Porphyromonas gingivalis (Pg) oral bacteria by monitoring their metabolomic evolution using in situ Raman spectroscopy. Pg-OMVs are a key factor in Alzheimer's disease (AD) pathogenesis, as they act as efficient vectors for the delivery of toxins promoting neuronal damage. However, the chemical mechanisms underlying the direct impact of Pg-OMVs on cell metabolites at the molecular scale still remain conspicuously unclear. A widely used in vitro model employing neuroblastoma SH-SY5Y cells (a sub-line of the SK-N-SH cell line) was spectroscopically analyzed in situ before and 6 h after Pg-OMV contamination. Concurrently, Raman characterizations were also performed on isolated Pg-OMVs, which included phosphorylated dihydroceramide (PDHC) lipids and lipopolysaccharide (LPS), the latter in turn being contaminated with a highly pathogenic class of cysteine proteases, a key factor in neuronal cell degradation. Raman characterizations located lipopolysaccharide fingerprints in the vesicle structure and unveiled so far unproved aspects of the chemistry behind protein degradation induced by Pg-OMV contamination of SH-SY5Y cells. The observed alterations of cells' Raman profiles were then discussed in view of key factors including the formation of amyloid ß (Aß) plaques and hyperphosphorylated Tau neurofibrillary tangles, and the formation of cholesterol agglomerates that exacerbate AD pathologies.

Instantaneous Inactivation of Herpes Simplex Virus by Silicon Nitride Bioceramics.

Hydrolytic reactions taking place at the surface of a silicon nitride (Si3N4) bioceramic were found to induce instantaneous inactivation of Human herpesvirus 1 (HHV-1, also known as Herpes simplex virus 1 or HSV-1). Si3N4 is a non-oxide ceramic compound with strong antibacterial and antiviral properties that has been proven safe for human cells. HSV-1 is a double-stranded DNA virus that infects a variety of host tissues through a lytic and latent cycle. Real-time reverse transcription (RT)-polymerase chain reaction (PCR) tests of HSV-1 DNA after instantaneous contact with Si3N4 showed that ammonia and its nitrogen radical byproducts, produced upon Si3N4 hydrolysis, directly reacted with viral proteins and fragmented the virus DNA, irreversibly damaging its structure. A comparison carried out upon testing HSV-1 against ZrO2 particles under identical experimental conditions showed a significantly weaker (but not null) antiviral effect, which was attributed to oxygen radical influence. The results of this study extend the effectiveness of Si3N4's antiviral properties beyond their previously proven efficacy against a large variety of single-stranded enveloped and non-enveloped RNA viruses. Possible applications include the development of antiviral creams or gels and oral rinses to exploit an extremely efficient, localized, and instantaneous viral reduction by means of a safe and more effective alternative to conventional antiviral creams. Upon incorporating a minor fraction of micrometric Si3N4 particles into polymeric matrices, antiherpetic devices could be fabricated, which would effectively impede viral reactivation and enable high local effectiveness for extended periods of time.

Effect of Carbonate Source on the Dehydrofluorination Process in Polyvinylidene Fluoride/Alkali Metal Carbonate Composites.

In this paper, Raman and X-ray photoelectron spectroscopies were applied to analyze compositional and structural variations of the generated activated carbon (AC), as induced by changing carbonate source in three different types of systems, PVDF/M2CO3 (M = Li, Na, and K). According to the variations of I D/I G and sp2/sp3 ratios, a strong dependence of the AC structure on the type and content of the initial carbonate was found, determined by practical dehydrofluorination reactions associated with oxygen incorporation in AC and side reactions, because of the property variation induced by the difference in the cation of the carbonate sources. This procedure clarified the process of PVDF dehydrofluorination and the formation of activated carbon, which helps to optimize the material performance of the percolative composite for flexible energy storage applications.

Design of Size-Controlled Sulfur Nanoparticle Cathodes for Lithium-Sulfur Aviation Batteries.

Lithium-sulfur (Li-S) battery has been considered as a strong contender for commercial aerospace battery, but the commercialization requires Ah-level pouch cells with both efficient discharge at high rates and ultra-high energy density. In this paper, the application of lithium-sulfur batteries for powering drones by using the cathode of highly dispersed sulfur nanoparticles with well-controlled particle sizes have been realized. The sulfur nanoparticles are prepared by a precipitation method in an eco-friendly and efficient way, and loaded on graphene oxide-cetyltrimethylammonium bromide by molecular grafting to realize a large-scale fabrication of sulfur-based cathodes with superior electrochemical performance. A button cell based on the cathode exhibits an excellent discharge capacity of 62.8 mAh cm-2 at a high sulfur loading of 60 mg cm-2 (i.e., 1046.7 mAh g-1 ). The assembled miniature pouch cell (PCmini) shows a discharge capacity of 130 mAh g-1 , while the formed Ah-level pouch cell (PCAh) achieves energy density of 307 Wh kg-1 at 0.3C and 92 Wh kg-1 at 4C. Especially, a four-axis propeller drone powered by the PC has successfully completed a long flight (>3 min) at high altitudes, demonstrating the practical applicability as aviation batteries.

In Situ Raman Analysis of Biofilm Exopolysaccharides Formed in Streptococcus mutans and Streptococcus sanguinis Commensal Cultures.

This study probed in vitro the mechanisms of competition/coexistence between Streptococcus sanguinis (known for being correlated with health in the oral cavity) and Streptococcus mutans (responsible for aciduric oral environment and formation of caries) by means of quantitative Raman spectroscopy and imaging. In situ Raman assessments of live bacterial culture/coculture focusing on biofilm exopolysaccharides supported the hypothesis that both species engaged in antagonistic interactions. Experiments of simultaneous colonization always resulted in coexistence, but they also revealed fundamental alterations of the biofilm with respect to their water-insoluble glucan structure. Raman spectra (collected at fixed time but different bacterial ratios) showed clear changes in chemical bonds in glucans, which pointed to an action by Streptococcus sanguinis to discontinue the impermeability of the biofilm constructed by Streptococcus mutans. The concurrent effects of glycosidic bond cleavage in water-insoluble α - 1,3-glucan and oxidation at various sites in glucans' molecular chains supported the hypothesis that secretion of oxygen radicals was the main "chemical weapon" used by Streptococcus sanguinis in coculture.

Effect of Ga2O3 Dopant on High Speed Sintered 5 mol% Y2O3 Stabilized Dental Zirconia.

The high-speed sintering of zirconia has become essential for ceramic dental prosthesis treatment in a single visit. Previous studies have shown that 5 mol% yttria-stabilized zirconia (5Y zirconia), with the exception of some types, loses strength and translucency with high-speed sintering. In this study, 0.15-0.92 wt% Ga2O3, which is expected to promote the sintering of zirconia, was added to improve the properties of 5Y zirconia high-speed sintered bodies, and the effect of its addition was evaluated. The specimens were characterized by density and translucency measurements, a three-point bending test, X-ray diffraction (XRD), scanning electron microscopy (SEM), and shrinkage measurement. The addition of Ga2O3 improved both translucency and flexural strength of 5Y zirconia high-speed sintered bodies. XRD and SEM observations revealed that this improvement in properties was due to the change in the crystal phase composition and the decrease in the amount and size of pores due to the addition of Ga2O3. Shrinkage measurements also revealed that the addition of Ga2O3 changed the sintering behavior of 5Y zirconia, suggesting that this change led to a reduction in porosity. From the above results, it was concluded that Ga2O3 addition is effective in improving the properties of 5Y zirconia high-speed sintered bodies.

Facile syntheses of Fe2O3-rGO and NiCo-LDH-rGO nanocomposites for high-performance electrochemical capacitors.

Faraday-type electrode materials and devices for electrochemical capacitors have been widely investigated. However, their applications are severely limited by the preparation method and cost of electrode materials. In this work, high-performance electrochemical capacitors were successfully assembled using Fe2O3-decorated reduced graphene oxide (rGO) nanocomposites and NiCo-Layered Double Hydroxides (LDH) as the anode and cathode, respectively. An easy and efficient approach (the modified precipitation method) for the large-scale fabrication was used to prepare Fe2O3 and NiCo-LDH, supported by rGO sheets, respectively. The anode material, Fe2O3-rGO, exhibited an excellent specific capacitance (Csp) of 1073 F g-1 at a current density of 1 A g-1 and a retention rate of 92 % at 10 A g-1, while the NiCo-LDH-rGO cathode material provided a Csp of 1850 F g-1 at 1 A g-1 and maintained 84 % at 10 A g-1. The effective combination of these electrodes for the NiCo-LDH-rGO//Fe2O3-rGO electrochemical capacitors resulted in an excellent energy density of 108 Wh/kg at a power density of 884 W/kg, with remarkable cycling stability (80 % after 1000 cycles at 10 A g-1). We believe that this work, including the proposed method and electrode materials, will advance the further development and commercialization of electrochemical capacitors.

Osseointegration Properties of Titanium Implants Treated by Nonthermal Atmospheric-Pressure Nitrogen Plasma.

Pure titanium is used in dental implants owing to its excellent biocompatibility and physical properties. However, the aging of the material during storage is detrimental to the long-term stability of the implant after implantation. Therefore, in this study, we attempted to improve the surface properties and circumvent the negative effects of material aging on titanium implants by using a portable handheld nonthermal plasma device capable of piezoelectric direct discharge to treat pure titanium discs with nitrogen gas. We evaluated the osteogenic properties of the treated samples by surface morphology and elemental analyses, as well as in vitro and in vivo experiments. The results showed that nonthermal atmospheric-pressure nitrogen plasma can improve the hydrophilicity of pure titanium without damaging its surface morphology while introducing nitrogen-containing functional groups, thereby promoting cell attachment, proliferation, and osseointegration to some extent. Therefore, nitrogen plasma treatment may be a promising method for the rapid surface treatment of titanium implants.

Raman Analyses of Laser Irradiation-Induced Microstructural Variations in Synthetic Hydroxyapatite and Human Teeth.

The microstructural and molecular-scale variations induced by laser irradiation treatment on human teeth enamel in comparison with synthetic hydroxyapatite (HAp) were examined through Raman microprobe spectroscopy as a function of irradiation power. The results demonstrated that laser irradiation could modify stoichiometry, microstructure, and the population of crystallographic defects, as well as the hardness of the materials. These modifications showed strong dependences on both laser power and initial nonstoichiometric structure (defective content of HPO4), because of the occurrence of distinct reactions and structural reconstruction. The reported observations can redirect future trends in tooth whitening by laser treatment and the production of HAp coatings because of the important role of stoichiometric defects.

Raman Metabolomics of Candida auris Clades: Profiling and Barcode Identification.

This study targets on-site/real-time taxonomic identification and metabolic profiling of seven different Candida auris clades/subclades by means of Raman spectroscopy and imaging. Representative Raman spectra from different Candida auris samples were systematically deconvoluted by means of a customized machine-learning algorithm linked to a Raman database in order to decode structural differences at the molecular scale. Raman analyses of metabolites revealed clear differences in cell walls and membrane structure among clades/subclades. Such differences are key in maintaining the integrity and physical strength of the cell walls in the dynamic response to external stress and drugs. It was found that Candida cells use the glucan structure of the extracellular matrix, the degree of α-chitin crystallinity, and the concentration of hydrogen bonds between its antiparallel chains to tailor cell walls' flexibility. Besides being an effective ploy in survivorship by providing stiff shields in the α-1,3-glucan polymorph, the α-1,3-glycosidic linkages are also water-insoluble, thus forming a rigid and hydrophobic scaffold surrounded by a matrix of pliable and hydrated ß-glucans. Raman analysis revealed a variety of strategies by different clades to balance stiffness, hydrophobicity, and impermeability in their cell walls. The selected strategies lead to differences in resistance toward specific environmental stresses of cationic/osmotic, oxidative, and nitrosative origins. A statistical validation based on principal component analysis was found only partially capable of distinguishing among Raman spectra of clades and subclades. Raman barcoding based on an algorithm converting spectrally deconvoluted Raman sub-bands into barcodes allowed for circumventing any speciation deficiency. Empowered by barcoding bioinformatics, Raman analyses, which are fast and require no sample preparation, allow on-site speciation and real-time selection of appropriate treatments.

Evaluation of the Effect of High-Speed Sintering and Specimen Thickness on the Properties of 5 mol% Yttria-Stabilized Dental Zirconia Sintered Bodies.

High-speed sintering of zirconia has become essential to single-visit dental prosthetic treatments. This important prosthetic dentistry technique demands a translucent material tougher than porcelain. Previous studies on high-speed sintered zirconia did not take heat and material thickness into consideration. We evaluated pre-sintered specimen thickness and the effect of high-speed sintering on the properties of 5 mol% Y2O3-stabilized zirconia (5Y zirconia). High-speed sintered bodies of 5Y zirconia were evaluated by density measurements, translucency measurements, three-point flexural and fracture toughness tests, X-ray diffraction (XRD), and scanning electron microscopy (SEM). High-speed sintering reduced the translucency and mechanical properties of 5Y zirconia. XRD and SEM observation results clarified that these reductions were due to the change in crystal phase composition and to the increase in residual pores, respectively, both resulting from high-speed sintering. Moreover, in high-speed sintering, as the thickness of the specimen increased, the number and size of internal pores increased, and the translucency and strength decreased. The threshold value for avoiding a reduction in translucency and mechanical properties was found to lie at ~4.4 mm. From the above results, it was concluded that 5Y zirconia is not suitable for high-speed sintering applications.