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Gingival crevicular fluid (GCF) is a site-specific exudate deriving from the epithelium lining of the gingival sulcus. GCF analysis provides a simple and noninvasive diagnostic procedure to follow-up periodontal and bone remodeling in response to diseases or mechanical stimuli such as orthodontic tooth movement (OTM). In recent years, the use of vibrational spectroscopies such as Fourier Transform Infrared and Raman microspectroscopy and Surface-Enhanced Raman spectroscopy contributed to characterizing changes in GCF during fixed orthodontic treatment. Amide I band plays a relevant role in the analysis of these changes. The aim of this study was to investigate the spectroscopy response of Amide I depending on the OTM process duration. A model based on Gaussian-Lorentzian curves was used to analyze the infrared spectra, while only Lorentzian functions were used for Raman and SERS spectra. Changes induced by the OTM process in subcomponents of the Amide I band were determined and ascribed to secondary structure modification occurring in proteins. The vibrational spectroscopies allow us to efficiently monitor the effects of the orthodontic force application, thus gaining increasing attention as tools for individual patient personalization in clinical practice.
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Amidas , Líquido del Surco Gingival , Humanos , Amidas/análisis , Espectroscopía Infrarroja por Transformada de Fourier , Líquido del Surco Gingival/química , Técnicas de Movimiento Dental/métodos , EncíaRESUMEN
A main factor hampering life in space is represented by high atomic number nuclei and energy (HZE) ions that constitute about 1% of the galactic cosmic rays. In the frame of the "STARLIFE" project, we accessed the Heavy Ion Medical Accelerator (HIMAC) facility of the National Institute of Radiological Sciences (NIRS) in Chiba, Japan. By means of this facility, the extremophilic species Haloterrigena hispanica and Parageobacillus thermantarcticus were irradiated with high LET ions (i.e., Fe, Ar, and He ions) at doses corresponding to long permanence in the space environment. The survivability of HZE-treated cells depended upon either the storage time and the hydration state during irradiation; indeed, dry samples were shown to be more resistant than hydrated ones. With particular regard to spores of the species P. thermantarcticus, they were the most resistant to irradiation in a water medium: an analysis of the changes in their biochemical fingerprinting during irradiation showed that, below the survivability threshold, the spores undergo to a germination-like process, while for higher doses, inactivation takes place as a consequence of the concomitant release of the core's content and a loss of integrity of the main cellular components. Overall, the results reported here suggest that the selected extremophilic microorganisms could serve as biological model for space simulation and/or real space condition exposure, since they showed good resistance to ionizing radiation exposure and were able to resume cellular growth after long-term storage.
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Optical vibrational techniques show a high potentiality in many biomedical fields for their characteristics of high sensitivity in revealing detailed information on composition, structure, and molecular interaction with reduced analysis time. In the last years, we have used these techniques for investigating gingival crevicular fluid (GCF) and periodontal ligament (PDL) during orthodontic tooth treatment. The analysis with Raman and infrared signals of GCF and PDL samples highlighted that different days of orthodontic force application causes modifications in the molecular secondary structure at specific wavenumbers related to the Amide I, Amide III, CH deformation, and CH3/CH2. In the present review, we report the most relevant results and a brief description of the experimental techniques and data analysis procedure in order to evidence that the vibrational spectroscopies could be a potential useful tool for an immediate monitoring of the individual patient's response to the orthodontic tooth movement, aiming to more personalized treatment reducing any side effects.
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SIGNIFICANCE: A noninvasive method based on surface-enhanced Raman spectroscopy (SERS) of tears was proposed as a support for diagnosing neurodegenerative pathologies, including different forms of dementia and Alzheimer's disease (AD). In this field, timely and reliable discrimination and diagnosis are critical aspects for choosing a valid medical therapy, and new methods are highly required. AIM: The aim is to evince spectral differences in SERS response of human tears from AD affected, mild cognitive impaired (MCI), and healthy control (Ctr) subjects. APPROACH: Human tears were characterized by SERS coupled with multivariate data analysis. Thirty-one informed subjects (Ctr, MCI, and AD) were considered. RESULTS: Average SERS spectra from Ctr, MCI, and AD subjects evidenced differences related to lactoferrin and lysozyme protein components. Quantitative changes were also observed by determining the intensity ratio between selected bands. We also constructed a classification model that discriminated among AD, MCI, and Ctr subjects. The model was built using the scores obtained by performing principal component analysis on specific spectral regions (i-PCA). CONCLUSIONS: The results are very encouraging with interesting perspectives for medical applications as support of clinical diagnosis and discrimination of AD from other forms of dementia.
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Enfermedad de Alzheimer , Enfermedades Neurodegenerativas , Enfermedad de Alzheimer/diagnóstico por imagen , Femenino , Humanos , Masculino , Análisis Multivariante , Enfermedades Neurodegenerativas/diagnóstico por imagen , Análisis de Componente Principal , Espectrometría RamanRESUMEN
Monitoring the spore life cycle is one of the main issues in several fields including environmental control, sustainable ecosystems, food security, and healthcare systems. In this framework, the study of the living organism resistance to extreme conditions like those mimicking space environments is particularly interesting. The assessment of the local change of the pH level can be extremely useful for this purpose. An optical physiometer method based on the Raman response of the graphene, which is able to locally sense pH of a fluid on a micrometric scale, has been recently proposed. Due to the presence of π -bonds at the surface, the electronic doping of graphene is determined by the external conditions and can be electrochemically controlled or altered by the contact with an acid or alkaline fluid. The doping level affects the vibrational energies of the graphene that can be monitored by conventional Raman spectroscopy. In addition, Surface-Enhanced Raman Spectroscopy (SERS) can give direct information on the biochemical changes occurring in spore components. In this work, we propose the joint use of Graphene-Based Raman Spectroscopy (GbRS) and SERS for the monitoring of the response of spores to exposure to low temperatures down to 100 K. The spores of the thermophilic bacterium Parageobacillus thermantarcticus isolated from an active volcano of Antarctica (Mt. Melbourne) were investigated. These spores are particularly resistant to several stressing stimuli and able to adapt to extreme conditions like low temperatures, UV irradiation, and γ -rays exposure. The results obtained showed that the joint use of GbRS and SERS represents a valuable tool for monitoring the physio-chemical response of bacterial spores upon exposure to stressing stimuli.
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Espectrometría Raman , Regiones Antárticas , Bacillaceae , Ecosistema , Grafito , Esporas Bacterianas , TemperaturaRESUMEN
The aim of the study was to examine the biochemical and structural changes occurring in the periodontal ligament (PDL) during orthodontic-force application using micro-Raman spectroscopy ( µ -RS). Adolescent and young patients who needed orthodontic treatment with first premolar extractions were recruited. Before extractions, orthodontic forces were applied using a closed-coil spring that was positioned between the molar and premolar. Patients were randomly divided into three groups, whose extractions were performed after 2, 7, and 14 days of force application. From the extracted premolars, PDL samples were obtained, and a fixation procedure with paraformaldehyde was adopted. Raman spectra were acquired for each PDL sample in the range of 1000-3200 cm - 1 and the more relevant vibrational modes of proteins (Amide I and Amide III bands) and CH 2 and CH 3 modes were shown. Analysis indicated that the protein structure in the PDL samples after different time points of orthodontic-force application was modified. In addition, changes were observed in the CH 2 and CH 3 high wavenumber region due to local hypoxia and mechanical force transduction. The reported results indicated that µ -RS provides a valuable tool for investigating molecular interchain interactions and conformational modifications in periodontal fibers after orthodontic tooth movement, providing quantitative insight of time occurring for PDL molecular readjustment.
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Ligamento Periodontal , Espectrometría Raman/métodos , Técnicas de Movimiento Dental , Adolescente , Adulto , Femenino , Humanos , Masculino , Ligamento Periodontal/química , Ligamento Periodontal/fisiología , Proteínas/análisis , Proteínas/química , Adulto JovenRESUMEN
Tears are exceptionally rich sources of information on the health status of the eyes, as well as of whole body functionality, due to the presence of a large variety of salts and organic components whose concentration can be altered by pathologies, eye diseases and/or inflammatory processes. Surface enhanced Raman spectroscopy (SERS) provides a unique method for analyzing low concentrations of organic fluids such as tears. In this work, a home-made colloid of gold nanoparticles has been used for preparing glass substrates able to efficiently induce an SERS effect in fluid samples excited by a Heâ»Ne laser ( λ = 633 nm). The method has been preliminary tested on Rhodamine 6G aqueous solutions at different concentrations, proving the possibility to sense substance concentrations as low as few µ M, i.e., of the order of the main tear organic components. A clear SERS response has been obtained for human tear samples, allowing an interesting insight into tear composition. In particular, aspartic acid and glutamic acid have been shown to be possible markers for two important human tear components, i.e., lactoferrin and lysozyme.
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Técnicas Biosensibles/métodos , Espectrometría Raman/métodos , Lágrimas/química , Oro/química , Humanos , Lactoferrina/química , Nanopartículas del Metal/química , Muramidasa/químicaRESUMEN
Graphene provides a unique way of sensing the local pH level of substances on the micrometric scale, with important implications for the monitoring of cellular metabolic activities where proton excretion could occur. Accordingly, an innovative biosensing approach for the quantification of the pH value of biological fluids, to be used also with small amounts of fluids, was realized and tested. It is based on the use of micro-Raman spectroscopy to detect the modifications of the graphene doping level induced by the contact of the graphene with the selected fluids. The approach was preliminarily tested on aqueous solutions of known pH values. It was then used to quantify the pH values of cell culture media directly exposed to different doses of X-ray radiation and to media exposed to X-ray-irradiated cells. The Raman response of cells placed on graphene layers was also examined.
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Células/química , Células/efectos de la radiación , Medios de Cultivo/química , Medios de Cultivo/efectos de la radiación , Grafito/química , Espectrometría Raman/métodos , Rayos X , Humanos , Concentración de Iones de HidrógenoRESUMEN
Gingival crevicular fluid (GCF) is a site-specific exudate in the gingival sulcus. GCF composition changes in response to diseases or mechanical stimuli, such as those occurring during orthodontic treatments. Raman microspectroscopy (µ-RS) and surface-enhanced Raman spectroscopy (SERS) were adopted for a GCF analysis during different initial phases of orthodontic force application. GCF samples were pooled from informed patients using paper cones. SERS spectra were obtained from GCF extracted from these cones, whereas µ-RS spectra were directly acquired on paper cones without any manipulation. The spectral characteristics of the main functional groups and the changes in cytochrome, amide III, and amide I contributions were highlighted in the different phases of orthodontic treatment with both SERS and µ-RS analysis. µ-RS directly performed on the paper cones together with proper statistical methods can offer an effective approach for the development of a tool for monitoring the processes occurring during orthodontic treatments, which may help the clinician in the choice of type of treatment individually for each patient and accelerate and improve the orthodontic therapy.
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Ortodoncia/métodos , Espectrometría Raman , Líquido del Surco Gingival/diagnóstico por imagen , HumanosRESUMEN
We study the current-voltage characteristics of Fe(Se,Te) thin films deposited on CaF2 substrates in form of nanostrips (width w ~ λ, λ the London penetration length). In view of a possible application of these materials to superconductive electronics and micro-electronics we focus on transport properties in small magnetic field, the one generated by the bias current. From the characteristics taken at different temperatures we derive estimates for the pinning potential U and the pinning potential range δ for the magnetic flux lines (vortices). Since the sample lines are very narrow, the classical creep flow model provides a sufficiently accurate interpretation of the data only when the attractive interaction between magnetic flux lines of opposite sign is taken into account. The observed voltages and the induced depression of the critical current of the nanostrips are compatible with the presence of a low number ([Formula: see text]) magnetic field lines at the equilibrium, a strongly inhomogeneous current density distribution at the two ends of the strips and a reduced Bean Livingston barrier. In particular, we argue that the sharp corners defining the bridge geometry represent points of easy magnetic flux lines injection. The results are relevant for creep flow analysis in superconducting Fe(Se,Te) nanostrips.
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Surface-Enhanced Raman Spectroscopy (SERS) enables the investigation of samples with weak specific Raman signals, such as opaque samples, including fruit juices and pulp. In this paper, biological apple juices and apple/pear pulp have been studied in order to evidence the presence of fructose and pectin, which are components of great relevance for quality assessment of these kinds of products. In order to perform SERS measurements a low-cost home-made substrate consisting of a glass slide decorated with 30-nm-sized gold nanoparticles has been designed and used. By employing a conventional micro-Raman spectroscopy set-up and a suitable data treatment based on "wavelet" denoising algorithms and background subtraction, spectra of pectin and fructose with clear Raman features have been obtained. The results have confirmed the potential of SERS in the food industry for product characterization, also considering the low-cost and the relative ease of the fabrication process of the employed SERS substrate.
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Análisis de los Alimentos , Fructosa , Frutas , Oro , Nanopartículas del Metal , Pectinas , Espectrometría RamanRESUMEN
Different chemical formulations for the synthesis of highly intercalated graphite bisulfate have been tested. In particular, nitric acid, potassium nitrate, potassium dichromate, potassium permanganate, sodium periodate, sodium chlorate, and hydrogen peroxide have been used in this synthesis scheme as the auxiliary reagent (oxidizing agent). In order to evaluate the presence of delamination, and pre-expansion phenomena, and the achieved intercalation degree in the prepared samples, the obtained graphite intercalation compounds have been characterized by scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), X-ray powder diffraction (XRD), infrared spectroscopy (FT-IR), micro-Raman spectroscopy (µ-RS), and thermal analysis (TGA). Delamination and pre-expansion phenomena were observed only for nitric acid, sodium chlorate, and hydrogen peroxide, while the presence of strong oxidizers (KMnO4, K2Cr2O7) led to stable graphite intercalation compounds. The largest content of intercalated bisulfate is achieved in the intercalated compounds obtained from NaIO4 and NaClO3.
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A micro-Raman spectroscopy investigation has been performed in vitro on single human mammary epithelial cells after irradiation by graded x-ray doses. The analysis by principal component analysis (PCA) and interval-PCA (i-PCA) methods has allowed us to point out the small differences in the Raman spectra induced by irradiation. This experimental approach has enabled us to delineate radiation-induced changes in protein, nucleic acid, lipid, and carbohydrate content. In particular, the dose dependence of PCA and i-PCA components has been analyzed. Our results have confirmed that micro-Raman spectroscopy coupled to properly chosen data analysis methods is a very sensitive technique to detect early molecular changes at the single-cell level following exposure to ionizing radiation. This would help in developing innovative approaches to monitor radiation cancer radiotherapy outcome so as to reduce the overall radiation dose and minimize damage to the surrounding healthy cells, both aspects being of great importance in the field of radiation therapy.
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Mama/citología , Células Epiteliales/efectos de la radiación , Espectrometría Raman/métodos , Análisis de Varianza , Células Epiteliales/química , Femenino , Humanos , Análisis de Componente Principal , Dosis de Radiación , Radiación Ionizante , Sensibilidad y EspecificidadRESUMEN
Micro-Raman Spectroscopy is an efficient method for analyzing biological specimens due to its sensitivity to subtle chemical and structural changes. The aim of this study was to use micro-Raman spectroscopy to analyze chemical and structural changes in periodontal ligament after orthodontic force application and in gingival crevicular fluid in presence of periodontal disease. The biopsy of periodontal ligament samples of premolars extracted for orthodontic reasons and the gingival crevicular fluid samples collected by using absorbent paper cones; were analyzed by micro-Raman spectroscopy. Changes of the secondary protein structure related to different times of orthodontic force application were reported; whereas an increase of carotene was revealed in patients affected by periodontal inflammation.
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Líquido del Surco Gingival/química , Ligamento Periodontal/química , Proteínas/análisis , Proteínas/ultraestructura , Espectrometría Raman/métodos , Técnicas de Movimiento Dental , Adolescente , Adulto , Análisis del Estrés Dental , Femenino , Humanos , Masculino , Estructura Secundaria de Proteína , Adulto JovenRESUMEN
A simple approach for the bulk production of carbon nanoscrolls (CNSs) is described. This method is based on the application of shear-friction forces to convert graphite nanoplatelets into carbon nanoscrolls using a bi-axially oriented polypropylene (BOPP) surface. The combined action of shear and friction forces causes the exfoliation of graphite nanoplatelets and the simultaneous roll-up of graphite layers. Evidence of the CNS formation is given by optical microscopy, scanning electron microscopy, and transmission electron microscopy. These investigations reveal that the CNSs have a long tube-like and fusiform structure with a hollow core surrounded by few layers of graphene. Micro-Raman spectroscopy shows that the produced structures are not defect free, and optical spectroscopy reveals distinctive features due to the presence of two weak absorption bands at 224 and 324 nm.
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Micro-Raman spectroscopy is a very promising tool for medical applications, thanks to its sensitivity to subtle changes in the chemical and structural characteristics of biological specimens. To fully exploit these promises, building a method of data analysis properly suited for the case under study is crucial. Here, a linear or univariate approach using a R2 determination coefficient is proposed for discriminating Raman spectra even with small differences. The validity of the proposed approach has been tested using Raman spectra of high purity glucose solutions collected in the 600 to 1,600 cm(-1) region and also from solutions with two known solutes at different concentrations. After this validation step, the proposed analysis has been applied to Raman spectra from oral human tissues affected by Pemphigus Vulgaris (PV), a rare life-threatening autoimmune disease, for monitoring disease follow-up. Raman spectra have been obtained in the wavenumber regions from 1,050 to 1,700 cm(-1) and 2,700 to 3,200 cm(-1) from tissues of patients at different stages of pathology (active PV, under therapy and PV in remission stage) as confirmed by histopathological and immunofluorescence analysis. Differences in the spectra depending on tissue illness stage have been detected at 1,150-1,250 cm(-1) (amide III) and 1,420-1,450 cm(-1) (CH3 deformation) regions and around 1,650 cm(-1) (amide I) and 2,930 cm(-1) (CH3 symmetric stretch). The analysis of tissue Raman spectra by the proposed univariate method has allowed us to effectively differentiate tissues at different stages of pathology.
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Monitoreo Fisiológico/métodos , Espectrometría Raman/métodos , Estudios de Seguimiento , HumanosRESUMEN
A wavelet multi-component decomposition algorithm has been used for data analysis of micro-Raman spectra of blood serum samples from patients affected by pemphigus vulgaris at different stages. Pemphigus is a chronic, autoimmune, blistering disease of the skin and mucous membranes with a potentially fatal outcome. Spectra were measured by means of a Raman confocal microspectrometer apparatus using the 632.8 nm line of a He-Ne laser source. A discrete wavelet transform decomposition method has been applied to the recorded Raman spectra in order to overcome problems related to low-level signals and the presence of noise and background components due to light scattering and fluorescence. This numerical data treatment can automatically extract quantitative information from the Raman spectra and makes more reliable the data comparison. Even if an exhaustive investigation has not been done in this work, the feasibility of the follow-up monitoring of pemphigus vulgaris pathology has been clearly proved with useful implications for the clinical applications.
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Liquid samples of clarified apple and apricot juices at different productionstages were investigated using visible light micro-Raman spectroscopy in order to assessits potential in monitoring fruit juice production. As is well-known, pectin plays a strategicrole in the production of clarified juice and the possibility of using Raman for its detectionduring production was therefore evaluated. The data analysis has enabled the clearidentification of pectin. In particular, Raman spectra of apple juice samples from washedand crushed fruits revealed a peak at 845 cm-1 (typical of pectin) which disappears in theRaman spectra of depectinised samples. The fructose content was also revealed by thepresence of four peaks at 823 cm-1, 872 cm-1, 918 cm-1 and 975 cm-1. In the case of apricotjuice, several Raman fingerprints of ß-carotene at 1008, 1159 and 1520 cm-1 were alsohighlighted. Present results resulted interesting for the exclusive use of optical methods forthe quantitative determination of the above-mentioned substances in place of thebiochemical assays generally used for this purpose, which are time consuming and requiredifferent chemical reagents for each of them.