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In recent years, the increase in consumer interest towards simpler and authentic lifestyles has led to an explosive growth in the production and business of typical agri-food products and, among these, of wines and its derived beverages. With the aim of promoting a typical Italian beverage, the so-called “Vino di visciole” or “Visner”, listed in the national table of traditional agri-food products, the antioxidant and colour properties of fifteen samples from different provinces of the Marche region and obtained with different recipes were analysed. The “in vitro” total antioxidant activity (TAA) determined using ABTS assays, total phenolic content (TPC), total flavonoid content (TFC), total anthocyanins content (TAC), and colour (Somers assay) were measured. In addition, a spectroscopic FT-IR and UV-Vis analysis was carried out to analyse samples with multivariate techniques. The results showed that the production area, the recipe, and the type of cherries used to make the alcoholic beverage do not influence the antioxidant properties and the phytochemical contents of the samples. The multivariate treatment of the spectroscopic features (mainly UV-Vis) rather allowed the differentiation of samples with high antioxidant activity using easy and low-cost instrumental techniques that require little time and can be employed in routine analysis.

Production of cost-efficient composite materials with desired physicochemical properties from low-cost waste material is much needed to meet the growing needs of the industrial sector. As a step forward, the current study reports for the first time an effective utilization of industrial metal (inorganic) waste as well as fall leaves (organic waste), to produce three types of nanomaterials at the same time; “Titanium Doped Activated Carbon Nanostructures (Ti-ACNs)”, “Nanocellulose (NCel)”, and combination of both “Titanium Doped Activated Carbon Cellulose Nanocomposite (Ti-AC-Cel-NC)”. X-ray diffraction (XRD), transmission electron microscopy (TEM) and microanalysis (EDXS) measurements reveal that the Ti-ACNs material is formed by Ti-nanostructures, generally poorly crystalized but in some cases forming hexagonal Ti-crystallites of 15 nm, embedded in mutated graphene clouds. Micro- Fourier transform infrared spectroscopy (micro-FTIR) confirms that the chemical structure of NCel with bond vibrations between 1035 to 2917 cm−1 remained preserved during Ti-AC-Cel-NC formation. The prepared materials (Ti-ACNs, Ti-AC-Cel-NC) have demonstrated rapid removal of organic pollutants (Crystal Violet, Methyl Violet) from wastewater through surface adsorption and photocatalysis. In the first 20 min, Ti-ACNs have adsorbed ≈87% of the organic pollutants and further photocatalyzed them up to ≈96%. When Ti-ACNs are combined with NCel, their efficiency is increased of about four times. This performance originates from the adsorption by mutated graphene-like carbon and assisted photocatalysis by Ti nanostructures as well as the good supporting capacity of NCel for the homogenous Ti-ACNs distribution.

In the present study, the cytotoxic effects of 5-azacytidine on primary Oral Squamous Cell Carcinoma cells (OSCCs) from human biopsies, and on Cancer Stem Cells (CSCs) from the same samples, were investigated by an in vitro Fourier Transform InfraRed Microscospectroscopy (FTIRM) approach coupled with multivariate analysis. OSCC is an aggressive tumoral lesion of the epithelium, accounting for ~90% of all oral cancers. It is usually diagnosed in advanced stages, and this causes a poor prognosis with low success rates of surgical, as well as radiation and chemotherapy treatments. OSCC is frequently characterised by recurrence after chemotherapy and by the development of a refractoriness to some employed drugs, which is probably ascribable to the presence of CSCs niches, responsible for cancer growth, chemoresistance and metastasis. The spectral information from FTIRM was correlated with the outcomes of cytotoxicity tests and image-based cytometry, and specific spectral signatures attributable to 5-azacytidine treatment were identified, allowing us to hypothesise the demethylation of DNA and, hence, an increase in the transcriptional activity, together with a conformational transition of DNA, and a triggering of cell death by an apoptosis mechanism. Moreover, a different mechanism of action between OSSC and CSC cells was highlighted, probably due to possible differences between OSCCs and CSCs response.

The polishing procedure is commonly performed after direct composite restorations, and little information exists regarding the right timing during which it should be performed on bulk fill composites. This study investigated the effect of polishing timing on the degree of conversion (DC), Vickers microhardness (VMH), and surface morphology of a methacrylate- (MET-) and dimethacrylate- (DMET-) based bulk fill composite, by using FT-NIR, microhardness tester, and SEM. Composite samples were divided as follows: in Group I (immediate), samples were polished immediately after curing (t0); in Group D (delayed), samples were polished after 24 h from curing (t24), whereas the unpolished samples were considered as controls (Group C). The DC and VMH values were evaluated before and after polishing, at t0 and t24. Statistical analysis was performed with a significance level set at p<0.05. At t0, DC increased after polishing in both tested composites (p<0.05), while at t24, Group I and Group D were not different. By considering VMH, in the case of MET, all groups were not different both at t0 and t24. On the other hand, at t0, VMH values of DMET increased after polishing. At t24, DMET Group I and DMET Group D were not different. Qualitative evaluations of scanning electron micrographs showed that the surface morphology of MET presented a more irregular aspect than the DMET one. In summary, since the immediate polishing of MET can improve the DC, without negatively affecting VMH, but showing an irregular surface, it is suggested to wait 24 hours before proceeding with polishing. Otherwise, for DMET, the immediate polishing could definitively be recommended, since it improves both DC and VMH, also producing a regular surface. Therefore, clinicians may always safely polish a restoration performed using DMET-based bulk fill composites in one-chair appointment, avoiding a second appointment.

Oral Squamous Cells Carcinoma (OSCC) is characterised by the risk of recurrence and the onset of a refractoriness response to chemotherapy drugs. These phenomena have been recently related to a subpopulation of Cancer Stem Cells (CSCs), which have either an innate or acquired drug resistance, triggered by chemotherapy treatments. In this light, to precisely target chemotherapy regimens, it is essential to improve knowledge on CSCs, with a particular focus on their molecular features. In this work, a subpopulation of CSCs, isolated by tumour sphere formation from primary OSCC cells, were treated with cisplatin for 16, 24 and 48 hours and analysed by infrared absorption and Raman microspectroscopies. CSC spectral data were compared with those obtained in previous work, for primary OSCC cells treated under the same conditions. Routine viability/apoptosis cell-based assays evidenced in CSCs and primary OSCCs, a similar degree of sensitivity to the drug at 24 hours, while a reversion of the conventional monotonic time response exhibited by OSCCs was shown by CSCs at 48 hours. This peculiar time response was also supported by the analysis of IR and Raman data, which pinpointed alterations in the lipid composition and DNA conformation in CSCs. The results obtained suggest that CSCs, although sharing with OSCC cells a similar sensitivity to cisplatin, display the onset of a mechanism of chemoresistance and enrichment of resistant CSCs as a result of drug treatment, shedding new light on the severe issue of refractoriness of some patients to chemotherapy conventionally used for OSCC.

Fourier-transform infrared hyperspectral imaging (FTIR-HSI) provides hyperspectral images containing both morphological and chemical information. It is widely applied in the biomedical field to detect tumor lesions, even at the early stage, by identifying specific spectral biomarkers. Pancreatic neoplasms present different prognoses and are not always easily classified by conventional analyses. In this study, tissue samples with diagnosis of pancreatic ductal adenocarcinoma and pancreatic neuroendocrine tumor were analyzed by FTIR-HSI and the spectral data compared with those from healthy and dysplastic samples. Multivariate/univariate approaches were complemented to hyperspectral images, and definite spectral markers of the different lesions identified. The malignant lesions were recognizable both from healthy/dysplastic pancreatic tissues (high values of phospholipids and triglycerides with shorter, more branched and less unsaturated alkyl chains) and between each other (different amounts of total lipids, phosphates and carbohydrates). These findings highlight different metabolic pathways characterizing the different samples, well detectable by FTIR-HSI.

In the present study, human primary oral squamous carcinoma cells treated with cisplatin and 5-fluorouracil were analyzed, for the first time, by in vitro FTIR Microspectroscopy (FTIRM), to improve the knowledge on the biochemical pathways activated by these two chemotherapy drugs. To date, most of the studies regarding FTIRM cellular analysis have been executed on fixed cells from immortalized cell lines. FTIRM analysis performed on primary tumor cells under controlled hydrated conditions provides more reliable information on the biochemical processes occurring in in vivo tumor cells. This spectroscopic analysis allows to get on the same sample and at the same time an overview of the composition and structure of the most remarkable cellular components. In vitro FTIRM analysis of primary oral squamous carcinoma cells evidenced a time-dependent drug-specific cellular response, also including apoptosis triggering. Furthermore, the univariate and multivariate analyses of IR data evidenced meaningful spectroscopic differences ascribable to alterations affecting cellular proteins, lipids and nucleic acids. These findings suggest for the two drugs different pathways and extents of cellular damage, not provided by conventional cell-based assays (MTT assay and image-based cytometry).

Fourier-Transform Infrared microspectroscopy, a largely used spectroscopic technique in basic and industrial researches, offers the possibility to analyze the vibrational features of molecular groups within a variety of environments. In the bioclinical field, and, in particular, in the study of cells, tissues and biofluids, it could be considered a supporting objective technique able to characterize the biochemical processes involved in relevant pathologies, such as tumoral diseases, highlighting specific spectral markers associable with the principal biocomponents (proteins, lipids and carbohydrates). In this article, we review the applications of infrared spectroscopy to the study of tumoral diseases of oral cavity compartments with the aim to improve understanding of biological processes involved during the onset of these lesions and to afford to an early diagnosis. Spectral studies on mouth, salivary glands and oral cystic lesions, objectively discriminate normal from dysplastic and cancer states characterizing also the grading.

In the present paper, we compared the histopathological and vibrational analyses of different tissue sections of Oral Squamous Cell Carcinoma (OSCC) at various malignancy grades, in order to unambiguously identify them. To achieve reliable results, healthy and dysplastic samples were also taken into account. FT-IR microspectroscopy is considered an effective tool for studying different molecular structures occurring in tumoral tissues and offers an interesting alternative to detect biochemical changes in a non-subjective way. In particular, on an adequate number of tissue sections affected by three different grades of OSSC (well G1, moderately G2, and poorly G3 differentiated), as well as on dysplastic and healthy tissues (all obtained from surgical resection), the chemical maps were acquired on meaningful areas containing both epithelial and connective structures. The multivariate analysis (Hierarchical Cluster Analysis, HCA, and Principal Component Analysis, PCA), performed separately on epithelial and connective spectral data, afforded to a good segregation for the different morphological structures. By analysing the representative spectra of healthy, dysplastic and tumoral epithelia and connectives, modifications were pin-pointed in the position of bands and absorbance band ratios usually associated with carcinogenesis. Above all, the changes in the protein pattern (with modifications in the length of side chains and in secondary structures), and in carbohydrates and nucleic acids moieties were associated with specific spectral markers of this pathology. The vibrational investigation led to a satisfactory understanding of these lesions so contributing to an early diagnosis, when the sole morphological inspection may result troublesome.

OBJECTIVES Fourier Transform Infrared microspectroscopy let characterize the macromolecular composition and distribution of tissues and cells, by studying the interaction between infrared radiation and matter. Therefore, we hypothesize to exploit this analytical tool in the analysis of inflamed pulps, to detect the different biochemical features related to various degrees of inflammation. MATERIALS AND METHODS IR maps of 13 irreversible and 12 hyperplastic pulpitis, together with 10 normal pulps, were acquired, compared with histological findings and submitted to multivariate (HCA, PCA, SIMCA) and statistical (one-way ANOVA) analysis. The fit of convoluted bands let calculate meaningful band area ratios (means ± s.d., P

Melatonin is an important hormone for the synchronization of the reproductive response to appropriate environmental conditions in photoperiodic animals. Even if a large amount of scientific evidences supports its positive impact on oocyte quality and fertilization rate, the biochemical processes involved are not completely understood. Fourier transform infrared (FT-IR) imaging technique has been applied to analyze the variations induced by melatonin administration on both growth and maturation phases of zebrafish oocytes. Chemical maps were acquired on thin ovary sections, from which average spectra were extracted for each class of oocytes. All average spectra were submitted to multivariate analysis, that separated them into different clusters according to oocyte class and treatment. The vibrational analysis highlighted modifications both in the proteic pattern, with an increment in helical structures with respect to β-sheets (amide I and II bands), and in the aliphatic chains and phosphate moieties for oocytes deriving from melatonin treated females. In addition, real time-PCR and enzyme-linked immunosorbent assay (ELISA) analyses showed that the administration of this hormone favoured the synthesis of vitellogenin. The reduction in gene expression involved in preventing oocyte maturation, such as bone morphogenetic protein ( bmp15 ) and transforming growth factor ( tgfβ1 ), was also observed together with the increase of the luteinizing hormone receptor ( lhr ), involved in the induction of oocyte maturation.

The aim of this study was to verify the effects of probiotic Lactobacillus rhamnosus on zebrafish oocyte maturation using FPA (focal plane array) FTIR imaging together with specific biochemical assays (SDS-PAGE, real-time PCR and enzymatic assay). Oocyte growth is prevalently due to a vitellogenic process which consists of the hepatic synthesis of vitellogenin and its selective uptake during maturation. The administration of L. rhamnosus IMC 501 for 10 days induced chemical changes to oocyte composition, promoting the maturation process. Some interesting biochemical features, linked to protein secondary structure (amide I band) and to phospholipidic and glucidic patterns, were detailed by vibrational analysis. The spectro- scopic results were supported by the early increase of the lysosomal enzyme involved in the final oocyte maturation, the cathepsin L. This enzyme increases during follicle maturation, with the highest levels in class IV oocytes. In treated females, class III oocytes showed higher cathepsin L gene expression and enzymatic activity, with levels comparable to class IV oocytes isolated from controls; this can be related to the proteolytic cleavage of the higher molecular mass yolk protein components, as evidenced by SDS-PAGE.

Zebrafish oocytes growth causes relevant modifications in protein components; in fact, the proteolytic cleavage of vitellogenin, a large sex-specific phospholipoglycoprotein synthesized in the female liver, leads to three main yolk protein components (phosphovitin and lipovitellins 1, 2), present as a complex in the oocyte. FT-IR Microspectroscopy could have the potentiality of monitoring these biochemical changes during the maturation process. Representative spectra for I–II, IIIA, IIIB and IV classes oocytes (Hierarchical Clustering Analysis and Principal Component Analysis) were investigated to find specific vibrational patterns corresponding to different maturation degrees. On going from I–II to IV class oocytes, relevant spectral differences were found with III class exhibiting an intermediate spectroscopic behaviour. In particular, the increase of the convoluted band at 2925 cm−1 (CH2 and CH3 stretching modes), as well as of intensity band ratios at 2926/2954 cm−1 (νasym CH2/CH3), 2854/2873 cm−1 (νsym CH2/CH3) and 1452/1392 cm−1 ( δ CH 2 / 3 / ν sym COO - ), suggest longer lipidic chains; broadening of Amide I and II bands and absorptions at 1737 (νC O, phospholipids) and 1157 cm−1 (νC O and νC OH carbohydrates) in lipovitellin and vitellogenin spectra, are present in III and IV classes.

Microimaging Fourier transform infrared spectroscopy is able to monitor differentiation between normal and malignant tissues. All the specimens, previously submitted to histological analysis, displayed abnormal spectra compared with the corresponding normal tissues with changes in many diagnostic bands like those arising from phosphate, C–O and CH stretching vibrational modes. The comparison between cancer (K) and connective (C) spectra evidenced the following differences: in the v CH region 3000–2800 cm −1 no hypomethylation effect was evident in K; the convolution of the bands of connective indicated an expected higher membrane fluidity; in the neoplastic zone, Amide I and II modes showed convoluted bands with maxima at 1651 and 1547 cm −1 , respectively, indicating an α-helix conformation of proteins due to changes in the secondary structure proteins upon carcinogenesis. Other signature bands, such as the deformation O–P–O phosphate band at 965 cm −1 , suggested DNA conformational changes in solid cancer, infiltrating cancer and neoplasia in the region 1350–800 cm −1 . These characteristic bands have been monitored as a function of the degree of cancer progression. Chemometric methods, such as principal component analysis (PCA) and hierarchical clustering analysis (HCA) have been used in order to distinguish spectra of neoplastic and normal zones.

Micro-imaging infrared spectroscopy has been performed on atheromatous plaques in order to localize and characterize substances responsible for the cytotoxic effects that prevent macrophages clearance of lipidic and calcified materials. In plaques with different graded atherosclerotic lesions, infrared determinations allowed to visualize gruel and ceroid toxic components and variously calcified zones. Compare correlations let to visualize the progression of the lesion on going from the lumen to the outer media of the plaque.

FT-IR microimaging was performed on colon tissues with the aim to characterize spectral ‘markers’ to distinguish healthy from pathological tissues. Evidences of spectral peculiarities were mainly found in the finger print region even in the presence of a low grade adenocarcinoma. The occurrence of inflammation and necrotic states can also be demonstrated. Through statistical analysis as well as custom map procedures it was possible to reconstruct the topological distribution of different biochemical states and to verify results from the histopathological analysis. Preliminary results from FT-NIR analysis are in substantial agreement with those in the mid infrared region.

Fourier transform infrared imaging (FTIRI) is a powerful tool for analyzing biochemical changes in tumoral tissues. The head and neck region is characterized by a great variety of lesions, with different degrees of malignancy, which are often difficult to diagnose. Schneiderian papillomas are sinonasal benign neoplasms arising from the Schneiderian mucosa; they can evolve into malignant tumoral lesions (squamous cell carcinoma). In addition, they can sometimes be confused with the more common inflammatory polyps. Therefore, an early and definitive diagnosis of this pathology is mandatory. Progressing in our research on the study of oral cavity lesions, 15 sections consisting of inflammatory sinonasal polyps, benign Schneiderian papillomas, and sinonasal undifferentiated carcinomas were analyzed using FTIRI. To allow a rigorous description of these pathologies and to gain objective diagnosis, the epithelial layer and the adjacent connective tissue of each section were separately investigated by following a multivariate analysis approach. According to the nature of the lesion, interesting modifications were detected in the average spectra of the different tissue components, above all in the lipid and protein patterns. Specific band-area ratios acting as spectral markers of the different pathologies were also highlighted.

Objective To characterize from a vibrational point of view the alterations caused by aging on human oocytes. Design Reproductive biology. Setting Private assisted reproductive technology clinic, synchrotron beam line, and university infrared laboratory. Patient(s) Twenty women of different ages (30 ± 2 and 39 ± 2 years) selected on the basis of detailed inclusion criteria and submitted to controlled ovarian stimulation according to a specific protocol. Intervention(s) Collection of 68 supernumerary oocytes that were not used during the IVF cycle from the above cited consenting patients. Main Outcome Measure(s) Focal Plane Array Fourier transform infrared (FTIR) analysis of human oocytes. Result(s) Specific spectral differences were highlighted in the two experimental groups of oocytes. In particular, in oocytes of 39-year-old women, the occurrence of peroxidative processes and a decrease in the amount of carbohydrates were observed, together with alterations in the phospholipid membrane, proteic pattern, and nucleic acids content. Conclusion(s) For the first time, FTIR spectroscopy was applied to human oocytes, leading to strong evidence of damage from aging in the gametes of mature women, which could be related to a decline in reproductive function. All the information obtained may be considered useful to improve the scientific knowledge on human reproduction and to exploit new strategies for detecting oocyte aging.

This study has been undertaken to investigate spectral features of cystic lesions of the jaw with the aim to understand their tumoral progression and to evidence initial signals of neoplastic changes. Three important groups (according to the World Health Organization classification) representing inflammatory (radicular) and developmental (orthokeratinized odontogenic, OOC) cysts as well as keratocystic odontogenic tumor (KCOT) of the jaw have been studied by microimaging infrared spectroscopy. From the spectral data analysed with the multivariate pattern recognition procedure, representative spectra were isolated and used to build correlation maps to localize the biochemical components in the tissue. The procedure enabled to better understand spectral features of these classes of cysts and to discriminate tumoral from non-tumoral spectra through the analysis of ‘vibrational markers’. In KCOT, the correlation of calcium derivatives (in metaplastic or displastic bones) with the tumor has been highlighted, too. The distribution of various biochemical components in the tissues, achieved through correlation maps of representative spectra, resulted in satisfactory agreement with the histopathological analysis. ‘Spectral isolation’ of micro tumorigenic zones in some normal OOC cysts, demonstrated the potentiality of infrared spectroscopy to evidence the early stage of a hidden lesion.

Stem cells are a promising source in a wide range of medical fields, included bone tissue regeneration. In this study, we investigated the molecular structure of dental pulp stem cells (DPSCs), during their differentiation towards the osteoblastic phenotype. DPSCs, easily achievable even from human young adults, were isolated from dental pulp of included third molar teeth and then cultured with appropriate medium to induce osteoblast differentiation. The FT-IR analysis of undifferentiated and differentiated DPSCs well pin-pointed biological changes during the osteogenic differentiation, above all in the Amide I, II and III vibrational modes as well as in the phosphate stretching vibrations. The spectroscopic study was supported by reverse transcriptase-polymerase chain reaction (RT-PCR) for the expression of specific osteoblastic markers, such as type I collagen (Coll-I), alkaline phosphatase (ALP), osteocalcin (OCN) and runt-related transcription factor 2 (RUNX2).

On continuing our studies on head and neck neoplasia, specimens from salivary gland tumors have been explored by using infrared microimaging spectroscopy to discern healthy from neoplastic tissues. Samples with Warthin tumor, epithelial displasia, marginal B-cell lymphoma, low-grade adenocarcinoma, and adenoid cystic carcinoma pathologies have been investigated by using conventional light sources. Changes were monitored at the molecular level, probing spectral markers such as Amide I and II, phosphate, nucleic acids, and carbohydrates vibrational modes. In all cases, supervised and unsupervised spectral analyses resulted in satisfactory agreement with histopathological findings. Microsc. Res. Tech., 2009. © 2008 Wiley-Liss, Inc.

A FT-IR microspectroscopy study on reactive lymph nodes and non-Hodgkin lymphomas is reported. Mid infrared absorption spectra collected at diffraction limit spatial resolution from reactive and neoplastic lymph nodes resulted sufficiently different once analysed by multivariate pattern recognition analysis to distinguish tumoral from non tumoral samples. The potential of infrared spectroscopy as a post-operative screening is gained by the use of a multielement Focal Plane Array detector. Spectral differences between normal and malignant spectra were mainly in the methyl stretching and in the low frequency region.