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This study aims to compare the degree of conversion of two different curing protocols used during adhesive cementation. The following resin luting agents were tested: Hri Flow (MF) and pre-heated Hri Micerium (MH); light-cure Nexus Third Generation (NX3L) and dual-cure Nexus Third Generation (NX3D); dual cured RelyX Ultimate (RXU) and light-cure RelyX Veneers (RXL). For each tested material, ten samples were prepared and divided into two groups which had different curing protocols (P1 and P2): in P1, samples were cured for 40 s; in P2, samples were cured for 5 s, and then, after 20 s, cured again for additional 40 s. The degree of conversion (DC) was evaluated both during the first 5 min of the curing phase and after 1, 2, 7, 14 and 28 days (p = 0.05). Different trends were observed in DC values after 5 min by comparing P1 and P2. In both P1 and P2, DC decreased as follows, MH > MF > NX3L > RXL > RXU > NX3D. There were significant differences of DC values among all resin luting agents (p p p > 0.05). Both P1 and P2 protocols let achieve an acceptable DC after 28 days. The tested P2 can be safely used to lute indirect restorations, simplifying the removal of cement excesses.

We have developed a photo-mobile polymer film, that combines organic and inorganic materials, to allow for controlled motion that can be triggered by light or heat. Our film is made using recycled quartz and consists of two layers: a multi-acrylate polymer layer and a layer containing oxidized 4-amino-phenol and N-Vinyl-1-Pyrrolidinone. The use of quartz in our film also gives it a high temperature resistance of at least 350 °C. When exposed to heat, the film moves in a direction that is independent of the heat source, due to its asymmetrical design. Once the heat source is removed, the film returns to its original position. ATR-FTIR measurements confirm this asymmetrical configuration. This technology may have potential applications in energy harvesting, due to the piezoelectric properties of quartz.

The paper deals with effects of two different widespread extraction methods (conventional extraction and Soxhlet extraction) and four different pre-treatments (homogenization with pressure and with blades, sonication, and impact with glass spheres) on the extraction yields and properties of polyhydroxyalkanoate (PHA) extracted from biomass coming from an innovative process (short-cut enhanced phosphorus and PHA recovery) applied in a real wastewater treatment plant. The results show that the two different extraction processes affected the crystallization degree and the chemical composition of the polymer. On the other hand, the extractive yield was highly influenced by pre-treatments: homogenization provided a 15% more extractive yield than the others. Homogenization, especially at high pressure, proved to be the best pre-treatment also in terms of the purity, visual appearance (transparency and clearness), thermal stability, and mechanical performances of the obtained PHA films. All the PHA films begin to melt long before their degradation temperature (Td > 200 °C): this allows their use in the fields of extrusion or compression moulding. Synopsis Optimizing the extraction of PHAs from municipal wastewater gives a double beneficial environmental impact: wastewater treatment and circular bio-based carbon upgrade to biopolymers for the production of bioplastics and other intersectoral applications.

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.

The occurrence, fate and removal of microplastics (MPs) in a wastewater treatment plant (WWTP) in Central Italy were investigated together with their potential adverse effects on anaerobic processes. In the influent of the WWTP, 3.6 MPs.L-1 were detected that mostly comprised polyester fibers and particles in the shape of films, ranging 0.1-0.5 mm and made of polyethylene and polypropylene (PP). The full-scale conventional activated sludge scheme removed 86% of MPs, with the main reduction in the primary and secondary settling. MPs particles bigger than 1 mm were not detected in the final effluent and some loss of polymers types were observed. In comparison, the pilot-scale upflow granular anaerobic sludge blanket (UASB) + anaerobic membrane bioreactor (AnMBR) configuration achieved 94% MPs removal with the abatement of 87% of fibers and 100% of particles. The results highlighted an accumulation phenomenon of MPs in the sludge and suggested the need to further investigate the effects of MPs on anaerobic processes. Accordingly, PP-MPs at concentrations from 5 PP-MPs.gTS-1 to 50 PP-MPs.gTS-1 were spiked in the pilot-scale UASB reactor that was fed with real municipal wastewater, where up to 58% decrease in methanogenic activity was observed at the exposure of 50 PP-MPs.gTS-1. To the best of our knowledge, the presented results will be the first to report of PP-MPs inhibition on anaerobic processes., The authors would like to acknowledge Matthew Chistolini for his valuable contribution to the investigation of PP-MPs in the pilot-scale UASB operation. The authors also would like to thank the water utility Viva Servizi for their support throughout the study. This study was supported by the Polytechnic University of Marche - MICROWASTE Strategic Project 2017 and by the European Commission with the LIFE program - "LIFE BLUE LAKES" under the Grant Agreement No. LIFE18 GIE/IT/000813. Alessia Foglia kindly acknowledges the Fondazione Cariverona for funding her PhD scholarship.

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.

A remarkable advantage of clay tiles roof coverings in hot climates is the realization of a ventilated air layer between them and the roofing underlay that allows a natural and forced convection through the tiles joints and the channel from eaves to ridge, thus cooling the roof materials. However recently, in many countries, regulatory developments on buildings energy efficiency or buildings sustainability certification protocols are increasingly encouraging the use of alternative strategies, with the aim of reducing the urban heat island (UHI) effect and the buildings’ cooling consumptions. Among them, the use of ‘cool’ materials for roof covering. These mandatory or voluntary measures de facto push the construction products market towards specific directions, risking penalizing traditional components such as clay tiles. This article reports the results of experimental and numerical activities carried out in order to extensively characterize the optical properties of clay tile materials and investigate their impact, also coupled with above sheathing ventilation, on the thermal performance of a ventilated roof under warm-temperate climate. In the first phase of the research, the main optical properties of over 30 different clay products have been experimentally characterized in order to get a clear and extensive picture of such properties for the materials spread in the market. In a second phase, starting from the thermal data collected on an experimental real-scale building, a dynamic energy analysis tool was calibrated and used to perform simulations by varying the optical properties of the roof covering thus assessing the impact on the roof temperatures, also in comparison to a clay tiles roof. The results underline that the use of the above sheathing ventilation obtained through clay tiles is an effective strategy to reduce roof temperatures, even if covering materials are not qualified as ‘cool’, thus impacting on both UHI and indoor comfort.

The cumulative impact of microplastic and organic enrichment is still largely unknown. Here, we investigated the microplastic contamination, the organic enrichment and their effects on meiofaunal distribution and diversity in two islands of the Maldivian archipelago: one more pristine, and another strongly anthropized. Field studies were coupled with manipulative experiments in which microplastic polymers were added to sediments from the non-anthropized island (i.e., without organic enrichment) to assess the relative effect of microplastic pollution on meiofauna assemblages. Our results reveal that the impact of microplastic contamination on meiofaunal abundance and taxa richness was more significant in the anthropized island, which was also characterized by a significant organic enrichment. Meiofauna exposed experimentally to microplastic contamination showed: i) the increased abundance of opportunistic nematodes and copepods and ii) a shift in the trophic structure, increasing relevance in epistrate-feeder nematodes. Based on all these results, we argue that the coexistence of chronic organic enrichment and microplastics can significantly increase the ecological impacts on meiofaunal assemblages. Since microplastic pollution in the oceans is predicted to increase in the next decades, its negative effects on benthic biodiversity and functioning of tropical ecosystems are expected to worsen especially when coupled with human-induced eutrophication. Urgent actions and management plans are needed to avoid the cumulative impact of microplastic and organic enrichment.

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.

This paper investigates the durability of a low-cost construction system named HOMEDONE developed to realize affordable and also temporary housing solutions. The system is based on the assembly of 3D-reinforced EPS panels externally topped off with a multi-layer rendering system. Similar technologies showed durability issues, especially in hot climates, due to the thermal and hygrometric stresses of the thin finishing layers when coupled to thick EPS panels and exposed to extreme events. For this reason, in this work freeze–thaw and wet/drying-UV aging tests on HOMEDONE panels with different finishing systems have been carried out, monitoring macroscopic, microscopic (ATR-FT-IR analysis) and bond strength variations due to aging. Results have pointed out good mechanical properties of the system and only small color variations of the finishing layer due to UV cycles. Then, HOMEDONE can be considered as a durable option for affordable and temporary housing solutions.

The history of the Chemistry within the Engineering Faculty will be described, and the evolution of the research topics will be shown till the actual main research lines. The explanation of these topics will also take into account the direction of their possible development in the next future.

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).

A desirable goal of the aquaculture sector is to replace most of fish meal and fish oil with more sustainable, cost-effective, and environmental friendly ingredients ensuring fish health and welfare standards. Due to minimal environmental impact, compared with most conventional feed commodities, insects deserve a growing attention as candidate ingredients for aquafeeds. The present study investigated, for the first time, the possible application of a 100% insect diet in zebrafish larval rearing. Through a multidisciplinary approach, the major biological responses of fish to the new diets were assessed. Results of biometry, fatty acid composition, expression of genes involved in fish growth, stress response, lipid metabolism, chitinolytic activity, gut inflammation, and liver macromolecular composition suggested a possible application of insect larvae for zebrafish larval rearing. However, further studies are necessary to better understand the use of this insect species in the rearing of fish.

In the last few years, vibrational spectroscopies have been widely applied in biology and medicine, as a synergic support to commonly used analytical and diagnostic techniques. This review summarizes the relevant researches carried out by using FTIR and Raman spectroscopy on oviparous and mammalian gametes, including human ones.

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.

A biomimetic bone-like composite, made of self-assembled collagen fibrils and carbonate hydroxyapatite nanocrystals, has been performed by an electrochemically-assisted deposition on titanium plate. The electrolytic processes have been carried out using a single type I collagen molecules suspension in a diluted Ca(NO3)2 and NH4H2PO4 solution at room temperature and applying a constant current for different periods of time. Using the same electrochemical conditions, carbonate hydroxyapatite nanocrystals or reconstituted collagen fibrils coatings were obtained. The reconstituted collagen fibrils, hydroxyapatite nanocrystals and collagen fibrils/apatite nanocrystals coatings have been characterized chemically, structurally and morphologically, as well as for their ability to bind fibronectin (FN). Fourier Transform Infrared microscopy has been used to map the topographic distribution of the coating components at different times of electrochemical deposition, allowing to single out the individual deposition steps. Moreover, roughness of Ti plate has been found to affect appreciably the nucleation region of the inorganic nanocrystals. Laser scanning confocal microscopy has been used to characterize the FN adsorption pattern on a synthetic biomimetic apatitic phase, which exhibits a higher affinity when it is inter-grown with the collagen fibrils. The results offer auspicious applications in the preparation of medical devices such as biomimetic bone-like compositecoated metallic implants. _ 2007 Elsevier B.V. All rights reserved. Keywords: Hydroxyapatite–collagen coating; Electrochemically-assisted deposition; Micro-imaging FTIR spectroscopy; Laser scanning confocal microscopy; Biomimetic crystal growth; Fibronectin binding

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.

The development of new methods for the preparation of C-glycosyl beta-amino acid libraries with chemical and stereochemical diversity levels was investigated and the results are described herein. Two complementary one-pot three-component Mannich-type and Reformatsky-type synthetic strategies have been developed for the construction of chiral 3-amino propanoate fragments (eventually bis-substituted at C-2) directly linked to the anomeric carbon of pyranose and furanose residues. Both methods involved as the initial step the coupling of a sugar aldehyde to p-methoxybenzylamine but differed in the nucleophile (a d(2) synthon equivalent) which was successively added: a ketene silyl acetal (Mannich route) or a bromozinc enolate (Reformatsky route). Individual C-glycosyl beta-amino esters were isolated as single 3R diastereoisomers in fair to excellent yield (60-90%) and their structure assigned by NMR spectroscopy (Riguera protocol) supported by X-ray crystallography. A tentative explanation of the observed stereochemical outcome based on transition-state models is provided. A preliminary study on the synthesis of alpha,alpha-difluoro C-glycosyl beta-amino acids via a more traditional Reformatsky route is also reported.

A novel and versatile strategy for the synthesis of heterocyclic alpha-amino acids has been described. The use of components (aldehyde or beta-ketoester) bearing a masked glycinyl moiety in Biginelli and Hantzsch cyclocondensations allowed access to the 4-dihydropyrimidinyl-alpha-glycines, 4-dihydropyrimidinyl-alpha-alanines, 4-pyridyl-alpha-alanines, and 2-pyridyl-alpha-alanines classes. Dihydropyrimidinyl-amino acids were obtained as a mixture of diastereoisomers due to the formation of the stereocenter at C4 of the dihydropyrimidinone ring. Individual stereoisomers were isolated as pure compounds and their structures were assigned with the aid of X-ray crystallography and chiroptical properties. The enantiomeric purity of a representative selection of the above amino acids was greater than 96% as verified by derivatization to the corresponding Mosher's amides and subsequent (1)H and (19)F NMR spectroscopy. Incorporation of the 4-pyridyl-alpha-alanine derivative into a peptide chain is also described.

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.

There is an imperative need to distinguish stable from unstable and vulnerable plaques because of the occurrence of thrombosis following rupture of the plaque. Imaging techniques, invasive and noninvasive, are used routinely for the assessment of visualization and quantification of this pathology, even if invasive techniques cannot be applied in the screening of atherosclerotic plaques and noninvasive imaging has low reproducibility. The aim of this review was to verify the suitability and the advantages of using vibrational spectroscopy in the study of human atherosclerotic plaques. The utilization of validated vibrational techniques in clinical trials may be of great importance for the ability to diagnose the early onset of a disease, rapidly, noninvasively and unambiguously. These techniques, by using a worldwide accepted protocol in spectral investigation of bioclinical changes in the human body, have been successfully applied to assess vulnerable plaques in ex vivo and in vivo models, constituting a potentially useful support to classical clinical imaging techniques. It is conceivable that this fingerprinting approach, with further clinical validation, is ready to be developed for use and at the disposal of commercial and easy-to-use spectrometers, equipped with catheter-based systems for the in vivo imaging of atherosclerosis plaques.

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).

The Lewis acid catalyzed (BF 3 ·Et 2 O, CuCl, AcOH) one-pot three-component cyclocondensation of urea with C -glycosylated aldehydes and β-keto esters (Biginelli reaction) in a combinatorial manner afforded three different series of 3,4-dihydropyrimidin-2(1 H )-ones bearing C -glycosyl moieties at C4, C6, and at both C4 and C6.

The Yb(OTf) 3 catalyzed Biginelli cyclocondensation reaction of 3-hydroxybenzaldehyde, ethyl acetoacetate and thiourea afforded the corresponding dihydropyrimidine-2-thione, called monastrol, in 95% isolated yield. The chiral resolution of racemic monastrol, a mitosis blocker by kinesin Eg5 inhibition, was carried out on a preparative scale (ca. 100 mg) through diastereomeric N -3 ribofuranosyl amides.

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.