Uterine smooth muscle tumors include a broad range of neoplasms, from benign leiomyomas (LMs) to malignant leiomyosarcomas (LMS), as well as intermediate forms classified as Smooth Muscle Tumors of Uncertain Malignant Potential (STUMP). An accurate diagnosis of these tumor types is essential for their appropriate clinical management; however, it remains challenging due to possible overlapping of histological features. In this study, a multidisciplinary approach combining Fourier Transform Infrared Imaging (FTIRI) spectroscopy, a label-free and non-destructive analytical technique, with histology and statistical analyses have been exploited for investigating the morpho-chemical characteristics of these uterine smooth muscle tumors. The analysis aimed to identify new reliable and diagnostic spectral markers, complementary to traditional histology, and thus useful for improving accuracy in cases with uncertain morphological features. Tissue samples including different leiomyoma histological subtypes, such as usual, cellular, apoplectic, and bizarre, were analyzed and compared with LMS and healthy myometrium. The analysis of IR data, submitted to univariate and multivariate statistical approaches, such as Principal Component Analysis (PCA) and Hierarchical Cluster Analysis (HCA), revealed distinctive spectral profiles associated with each tumor type and indicated changes in collagen content and organization as key features for a reliable discrimination not only between benign and malignant tissues but also among different LM histotypes.

Objective:Plastics are widely used, including in medical applications. In dentistry, clear aligners (CAs) offer an alternative to fixed appliances, but their potential microplastics (MP) release raises health concerns. This study aims to investigate the detachment of MPs from CAs by analyzing the correlations between wearing time, manufacturing methods, and material composition. Methods:The following CAs were tested: Alleo (AL), F22 Aligner (F22), FlexiLigner (FL), Graphy (GP), Invisalign (INV), KeySplint (KS), Lineo (LIN), LuxCreo (LC), Spark (SP), and SureSmile (SS). One pair of CAs per group was immersed in artificial saliva, and the solution was stirred for 5 hours per day for 7 (T1) or 14 (T2) days to simulate mechanical friction. Saliva samples were subsequently vacuum-filtered through 1.6-μm pore-size membranes. Attenuated total reflectance-Fourier transform infrared spectroscopy was used to characterize the CA materials, and Raman microspectroscopy and scanning electron microscopy were used to assess the chemical composition, number, size, and shape of the MPs. The experiment was conducted in triplicate. Results:The release of MPs increased significantly from T1 to T2 (P < 0.05), especially those with a diameter < 5 μm. The mean size of the MPs did not significantly differ at either time point. MP release was associated with material composition at T1 and T2 (P < 0.0001). No significant correlation was observed with the manufacturing process, although GP, KS, and LC, which are three-dimensionally printed (3D-printed) CAs, released more uniform and spherical MPs. Conclusions:Replacing CAs after 7 days may help limit MP release, particularly smaller fragments; however, the clinical significance of this finding remains unclear and requires further in vivo investigation.

Cisplatin is a platinum-based chemotherapy drug with antimicrobial and antitumoral activity, largely used for a long time in the treatment of several cancers, including the Oral Squamous Cell Carcinoma (OSCC), which is one of the most frequent neoplasms of the oral cavity. Due to its aggressiveness and metastatic invasion, OSCC is characterized by poor outcome, often related also to chemoresistance mechanisms. The intracellular enzyme paraoxonase-2 (PON2) normally acts defending cells from the damages induced by Reactive Oxygen Species. Hence, in cancer cells, this enzyme can shield the potential of cisplatin, triggering a resistance mechanism. Based on this evidence, PON2 knockdown seems to be a valuable way to enhance the effects of chemotherapy, escaping this resistance. In this study, HOC621 and HSC-3 OSCC cell lines submitted to PON2 silencing were analyzed by Fourier Transform Infrared Microspectroscopy to evaluate the time-dependent changes occurring in these cells after cisplatin treatment. Spectral data were statistically analyzed by multivariate and univariate analyses and compared with MTT results. Positive feedback on cisplatin efficacy was found in both cell lines submitted to PON2 knockdown, even if with a different response. In particular, a less growth was found in PON2 silenced HOC 621 cells, respect to HSC-3 ones. Moreover, specific spectral markers (A 1172 /A TOT , A 1053 /A TOT , A 967 /A 1080 , and A 992 /A TOT band area ratios) were identified and statistically analyzed (p < 0.05): cellular alterations mainly in nucleic acids and carbohydrates were found in both cell lines, although more evident in HOC 621 ones, which therefore appeared to be more affected by chemotherapy treatment.

The mixing of Guanosine (Gua) and Guanosine 5'-monophosphate (GMP) in water in selected compositions yields highly hydrated, transparent, and self-healing self-assembled supramolecular G-hydrogels, attractive for biomedical applications. This work investigates how hydrogel composition affects solute transport, including diffusion, binding, loading, and release properties, using a set of fluorescent probes with varying size and polarity. Although small/wide-angle X-ray scattering techniques showed that no structural changes are induced by probe addition, even when intercalation into G-quadruplexes is expected, the internal mesh structure of the hydrogel, modulated by the Gua:GMP ratio, directly impacts probe diffusivity and loading. Tighter networks (e.g., 1:1) slow diffusion and enhance retention compared to looser configurations (e.g., 1:4). Moreover, UV-visible titrations revealed markedly different binding affinities (Kb ≈ 5.7 × 104 M-1 for DAPI, 8.0 × 103 M-1 for ThT, and 1.4 × 102 M-1 for RhB), which are expected to result in lower diffusion coefficients and slower release, especially for DAPI and ThT. Indeed, diffusion coefficients, obtained via fluorescence recovery after photobleaching and time-resolved fluorescence spectroscopy, reach 90, 20, and 60 μm2/s for FITC-dextran, ThT, and RhB, respectively. Probe release kinetics, modeled via Weibull fitting, indicated sustained release with characteristic times (τ) between 9.6 and 23.2 h and β ≈ 1 in 1× PBS, consistent with predominantly Fickian diffusion. Remarkably, switching to 10× PBS significantly accelerated release (τ reduced by ≈ 40-50%), suggesting that ionic strength and/or pH changes critically affect not only probe-hydrogel interactions but also the internal gel architecture, altering porosity, mesh size, and network tortuosity, thus enhancing molecular mobility. Overall, the G-hydrogel system offers a structurally tunable and composition-dependent platform capable of finely regulating molecular transport and release profiles, making it highly suitable for controlled drug delivery and adaptive biomaterial applications.

The Adriatic Sea is a critical neritic habitat for juvenile and adult female loggerhead sea turtles (Caretta caretta), where intense anthropogenic pressures and environmental stressors may influence their reproductive biology. Knowledge on the onset of puberty in this population is limited by scarce information on the sub-adult stage, a transitional phase in which reproductive competence is acquired. This study integrated histological analysis and Fourier-transform infrared (FTIR) imaging spectroscopy to provide both structural and biochemical characterization of folliculogenesis, with emphasis on vitellogenesis, in C. caretta from the north-central Adriatic Sea. Histological analysis determined the progression of follicle development, while FTIR imaging, a label-free and spatially resolved technique, mapped the distribution of proteins, lipids, and nucleic acids across ovarian compartments. Logistic regression estimated the size at which 50% of females are sexually mature (L50) at 58.54 cm Curved Carapace Length (CCL). Based on this value, 60% of sub-adult females were already mature, indicating earlier puberty than previously inferred from macroscopic criteria. These preliminary results, along with reports of sporadic nesting in the Adriatic, raise the question of whether this basin may host further nesting events in the future. FTIR imaging proved to be a powerful tool for reproductive biology in non-model marine vertebrates.

Sinus augmentation provides a well-established model for investigating the three-dimensional morphometry and macromolecular dynamics of bone regeneration, particularly when using biphasic calcium phosphate (BCP) graft substitutes. This case series included six biopsies from patients who underwent maxillary sinus augmentation using BCP granules composed of 30% hydroxyapatite (HA) and 70% β-tricalcium phosphate (β-TCP). Bone core biopsies were obtained at healing times of 6 months, 9 months, and 12 months. Histological evaluation yielded qualitative and quantitative insights into new bone distribution, while micro-computed tomography (micro-CT) and Raman microspectroscopy (RMS) were employed to assess the three-dimensional architecture and macromolecular composition of the regenerated bone. Micro-CT analysis revealed progressive maturation of the regenerated bone microstructure over time. At 6 months, the apical regenerated area exhibited a significantly higher mineralized volume fraction (58 ± 5%) compared to the basal native bone (44 ± 11%; p = 0.0170), as well as significantly reduced trabecular spacing (Tb.Sp: 187 ± 70 µm vs. 325 ± 96 µm; p = 0.0155) and degree of anisotropy (DA: 0.37 ± 0.05 vs. 0.73 ± 0.03; p < 0.0001). By 12 months, the mineralized volume fraction in the regenerated area (53 ± 5%) was statistically comparable to basal bone (44 ± 3%; p > 0.05), while Tb.Sp (211 ± 20 µm) and DA (0.23 ± 0.09) remained significantly lower (Tb.Sp: 395 ± 41 µm, p = 0.0041; DA: 0.46 ± 0.04, p = 0.0001), indicating continued structural remodelling and organization. Raman microspectroscopy further revealed dynamic macromolecular changes during healing. Characteristic β-TCP peaks (e.g., 1315, 1380, 1483 cm−1) progressively diminished over time and were completely absent in the regenerated tissue at 12 months, contrasting with their partial presence at 6 months. Simultaneously, increased intensity of collagen-specific bands (e.g., Amide I at 1661 cm−1, Amide III at 1250 cm−1) and carbonate peaks (1065 cm−1) reflected active matrix formation and mineralization. Overall, this case series provides qualitative and quantitative evidence that bone regeneration and integration of BCP granules in sinus augmentation continues beyond 6 months, with ongoing maturation observed up to 12 months post-grafting.

Background: The study exploited, for the first time, Attenuated Total Reflectance-Fourier Transform-InfraRed (ATR-FTIR) spectroscopy on human dental pulps at different timings of root resorption (RR) to deepen the biological mechanisms occurring in deciduous teeth (De) during their replacement with permanent ones. Methods: N:36 dental pulps from sound De were divided into the following: G0 (no RR); G1 (RR less than 1/3 of root length); G2 (RR not exceeding 2/3 of root length); and G3 (RR more than 2/3 of root length). Samples were analyzed by ATR-FTIR, and the spectral data were submitted to univariate (One-way ANOVA and Tukey’s multiple comparison tests; statistical significance set at p < 0.05) and multivariate (Principal Component Analysis, PCA) analyses. Results: PCA displayed good discrimination among groups, ascribable to: (i) the intensity of the peaks of nucleic acids (~1715 cm−1, ~1237 cm−1, ~964 cm−1, and ~815 cm−1) and carbohydrates (~1159 cm−1) which increased from G0 to G3 (p < 0.05); (ii) the relative amount of lipids which decreased from G0 to G3 (p < 0.05); and (iii) the intensity of the peaks at ~1014 cm−1, and ~875 cm−1 (phosphates and carbonates in hydroxyapatite), which decreased from G0 to G3 (p < 0.05). Conclusions: This study confirmed ATR-FTIR as a reliable and quick technique for the characterization of the dental pulp and highlighted a correlation between specific molecular changes in the dental pulp of deciduous teeth and different RR stages, shedding new light on this process and paving the way for future research, which could improve the clinical management of the primary dentition.

Guanosine nucleosides and nucleotides have the peculiar ability to self-assemble in water to form supramolecular complex architectures from G-quartets to G-quadruplexes. G-quadruplexes exhibit in turn a large liquid crystalline lyotropic polymorphism, but they eventually cross-link or entangle to form a densely connected 3D network (a molecular hydrogel), able to entrap very large amount of water (up to the 99% v/v). This high water content of the hydrogels enables tunable softness, deformability, self-healing, and quasi-liquid properties, making them ideal candidates for different biotechnological and biomedical applications. In order to fully exploit their possible applications, Attenuated Total Reflection-Fourier Transform InfraRed (ATR-FTIR) spectroscopy was used to unravel the vibrational characteristics of supramolecular guanosine structures. First, the characteristic vibrations of the known quadruplex structure of guanosine 5 '-monophosphate, '-monophosphate, potassium salt (GMP/K), were investigated: the identified peaks reflected both the chemical composition of the sample and the formation of quartets, octamers, and quadruplexes. Second, the role of K+ + and Na+ + cations in promoting the quadruplex formation was assessed: infrared spectra confirmed that both cations induce the formation of G-quadruplexes and that GMP/K is more stable in the G-quadruplex organization. Finally, ATR-FTIR spectroscopy was used to investigate binary mixtures of guanosine (Gua) and GMP/K or GMP/Na, both systems forming G-hydrogels. The same G-quadruplex-based structure was found in both mixtures, but the proportion of Gua and GMP affected some features, like sugar puckering, guanine vibrations, and base stacking, reflecting the known side-to-side aggregation and bundle formation occurring in these binary systems.

This review presents several alternatives to replace antibiotic therapy and make the European aquaculture industry more friendly and environmentally sustainable. The first part of this review highlights the growing importance of the aquaculture industry worldwide for its ability to supply low-cost proteins and lipids. The second part discusses different strategies for these replacements, from recombinant vaccines to diets with low environmental impact and rich in bioactive molecules that can benefit other species. Specifically, the beneficial effects of bioactive compounds present within insect meals are discussed. In addition, particular focus is placed on the importance of adopting sustainable protocols for fish farming, including supplements such as probiotics, prebiotics, and synbiotics capable of modulating the gut microbiota as the second brain. Those feed additives can stimulate European farmed species’ immunological systems, growth, and welfare.

Human pharmaceuticals represent a major challenge in natural environment. A better knowledge on their mechanisms of action and adverse effects on cellular pathways is fundamental to predict long-term consequences for marine wildlife. The FTIRI Imaging (FTIRI) spectroscopy represents a vibrational technique allowing to map specific areas of non-homogeneous biological samples, providing a unique biochemical and ultrastructural fingerprint of the tissue. In this study, FTIRI technique has been applied, for the first time, to characterize (i) the chemical building blocks of digestive glands of Mytilus galloprovincialis, (ii) alterations and (iii) resilience of macromolecular composition, after a 14-days exposure to 0.5 µg/L of carbamazepine (CBZ), valsartan (VAL) and their mixture, followed by a 14-days recovery period. Spectral features of mussels digestive glands provided insights on composition and topographical distribution of main groups of biological macromolecules, such as proteins, lipids, and glycosylated compounds. Pharmaceuticals caused an increase in the total amount of protein and a significant decrease of lipids levels. Changes in macromolecular features reflected the modulation of specific molecular and biochemical pathways thus supporting our knowledge on mechanisms of action of such emerging pollutants. Overall, the applied approach could represent an added value within integrated strategies for the effects-based evaluation of environmental contaminants.

Background One of the main factors for the osseointegration of dental implants is the development of an adequate soft tissue barrier, mainly composed by collagen, which protects the implant from bacterial development. The structural features of the peri-implant collagen are influenced by the implant components and, in particular, by the type of the surface. In the clinical practice, healing abutments are characterized by smooth surfaces, named machined. Recently, a new laser technique, Synthegra, has been developed to obtain a topography-controlled surface with micrometric regular pores that seems reducing the risk of peri-implantitis. Based on this background, this study aims investigating the structural organization and spatial distribution of collagen surrounding healing abutments characterized by laser-treated and machined surfaces. Methods Gingiva portions surrounding custom-made healing abutments (HA), characterized by alternated laser-treated and machined surfaces, were collected and analyzed by combining Fourier Transform InfraRed Imaging (FTIRI) spectroscopy, a non-invasive and high-resolution bidimensional analytical technique, with histological and multivariate analyses. Results Masson’s trichrome staining, specific for collagen, highlighted a massive presence of collagen in all the analyzed samples, evidencing a surface-related spatial distribution. The nature of collagen, investigated by the FTIRI spectroscopy, appeared more abundant close to the laser-treated surface, with a perpendicular disposition of the bundles respect to the HA; conversely, a parallel distribution was observed around the machined surface. A different secondary structure was also found, with a higher amount of triple helices and a lower quantity of random coils in collagen close to the laser treated surfaces. Conclusions FTIRI spectroscopy demonstrates that the use of a laser treated transmucosal surface can improve the morphological organization of the peri-implant collagen, which presents a distribution more similar to that of natural teeth. Trial registration : This trial is registered with ClinicalTrials.gov Identifier: (Registration Number: NCT05754970). Registered 06/03/2023, retrospectively registered, https://clinicaltrials.gov/show/NCT05754970 .

A crucial issue in forensics is determining the post-mortem interval (PMI), the time between death and the finding of a body. Despite various methods already employed for its estimation, only approximate values are currently achievable. Vitreous humor (VH) is an avascular tissue between the lens and the retina, mainly composed by a collagen fibers network, hyaluronic acid, and hyalocytes. Recently, it has received interest in forensic medicine, being easy to collect and susceptible to low microbiological contamination and putrefaction. Based on this evidence and thanks to the ability of Attenuated Total Reflectance - Fourier Transform InfraRed (ATR-FTIR) spectroscopy to perform fast analyses on a minimal sample amount, in this study, a new analytical approach to reliably estimate PMI is proposed combining ATR-FTIR analysis of VH human samples with multivariate statistical procedures, such as Principal Component Analysis (PCA) and Partial Least Squares-Discriminant Analysis (PLS-DA), for discriminant classification. Regression procedures, including Partial Least Squares Regression (PLSR), were performed: extremely positive results were obtained, and the most discriminant spectral features were highlighted (peaks at 1665, 1630, 1585, 1400, 1220, 1200, 1120, 854, 835, and 740 cm-1) and associated to PMI classes (average accuracy over 80 %). Specific and reliable markers able to correlate the macromolecular composition of VH with the PMI were identified, revealing a post-mortem protein degradation and amino acids deamination (decrease of proteins and increase of free amino acids and NH3), an increase of lactate, which diffuses from the retina to the VH, and changes in the hyaluronic acid component.

The increasing global concern over microplastic pollution has driven a surge in research efforts aimed at detecting microplastics across various ecosystems. Airborne microplastics (MPs) have been identified in remote environments worldwide, including Antarctica. However, data on bulk atmospheric deposition remain scarce. From January to December 2020, atmospheric deposition was directly collected using passive samplers placed in eight sites across Victoria Land. Using Raman Microspectroscopy, MPs were identified in six out of the seven samples collected (one sample was lost due to the extreme weather conditions). The average daily MP deposition for Victoria Land was 1.7 +/- 1.1 MPs m(-2) d(-1), with values ranging from 0.76 to 3.44 MPs m(-2) d(-1). The majority (53 %) of MPs found in the atmospheric deposition were in the size class of 5-10 mu m, and the main shape of MPs was fragments (95 %). The predominant plastic type was polypropylene (31 %), followed by polyethylene (19 %) and polycarbonate (12 %). Polystyrene, polyester, styrene and polyethylene terephthalate each accounted for similar to 6 %. Microplastics identified in the coastal sites may have local origins, potentially associated with scientific activities at research stations. Conversely, a backward trajectories analysis suggested a potential contribution of atmospheric transport to microplastic deposition at Larsen Glacier and Tourmaline Plateau, the two most remote sites of the study area, where the highest MP concentrations were detected. Our findings present the first evidence of microplastics in the Antarctic atmospheric deposition directly collected via passive samplers, highlighting the need for continued monitoring and research to assess the environmental impact of MPs, particularly in sensitive and remote ecosystems like Antarctica.

High resolution analysis of collagen bundles could provide information on tumor onset and evolution. This study was focused on the microarchitecture and biomolecular organization of collagen bundles in oral tongue squamous cell carcinoma (OTSCC). Thirty-five OTSCC biopsy samples were analyzed by synchrotron-based phase-contrast microcomputed tomography and Fourier transform infrared imaging (FTIRI) spectroscopy. PhC-microCT evidenced the presence of reduced and disorganized collagen in the tumor area compared to the extratumoral (ExtraT) one. FTIRI also revealed a reduction of folded secondary structures in the tumor area, and highlighted differences in the peritumoral (PeriT) areas in relation with the OTSCC stage, whereby a significantly lower amount of collagen with less organized fibers was found in the PeriT stroma of advanced-OTSCC stages. Interestingly, no significant morphometrical mismatches were detected in the same region by PhC-microCT analysis. These results suggest that biomolecular alterations in the OTSCC stroma temporally anticipate structural modifications of collagen bundle microarchitecture.

ATR-FTIR (Attenuated Total Reflectance Fourier Transform InfraRed) spectroscopy, combined with chemometric, represents a rapid and reliable approach to obtain information about the macromolecular composition of food and plant materials. With a single measurement, the chemical fingerprint of the analyzed sample is rapidly obtained. Hence, this technique was used for investigating 13 differently processed tea leaves (green, black and white) all grown and processed in European tea gardens, and their vacuum-dried tea brews, prepared using both hot and cold water, to observe how the components differ from tea leaf to the in-cup infusion. Spectra were collected in the 1800–600 cm−1 region and were submitted to Principal Component Analysis (PCA). The comparison of the spectral profiles of leaves and hot and cold infusions of tea from the same country, emphasizes how they differ in relation to the different spectral regions. Differences were also noted among the different countries. Furthermore, the changes observed (e.g., at ~1340 cm−1) due to catechin content, confirm the antioxidant properties of these teas. Overall, this experimental approach could be relevant for rapid analysis of various tea types and could pave the way for the industrial discrimination of teas and of their health properties without the need of time-consuming, lab chemical assays.

In this preliminary study, a multidisciplinary method based on high-resolution analytical techniques (such as microcomputed tomography, Raman Microspectroscopy, scanning electron microscopy, and Vickers microhardness test) was exploited to evaluate the alterations that occur in human teeth at the initial stage of the carious lesion. To this purpose, six extracted molars displaying a natural white spot lesion (WSL) were investigated. Specific morphological, structural, and chemical parameters, such as the mineral density, indentation hardness, molecular and elemental composition, and surface micromorphology were obtained on the WSL, and the results were statistically compared (t-test, p < 0.05) to those of the sound enamel on the same tooth. In the WSL, with respect to the sound area, a decrease in the mineral density and crystallinity was detected together with differences in the molecular composition and surface microstructure, such as the occurrence of micropores and irregularities. Moreover, the elemental analysis highlighted in WSL showed a statistically significant decrease in Ca and P percentages. In conclusion, this multidisciplinary approach allows us to fully characterize the area of interest, providing a deeper knowledge of these enamel lesions, which could have important clinical implications.

Multinuclear complexes are metal compounds featured by adjacent bound metal centers that can lead to unconventional reactivity. Some M2L4-type paddlewheel dinuclear complexes with monoanionic bridging ligands feature promising properties, including therapeutic ones. Molybdenum has been studied for the formation of multiple-bonded M2+ compounds due to their unique scaffold, redox, and spectroscopic properties as well as for applications in several fields including catalysis and biology. These latter are much less explored and only sporadic studies have been carried out. Here, a series of four dimolybdenum (II,II) carboxylate paddlewheel complexes were synthesized using different Non-Steroidal Anti-Inflammatory Drugs (NSAIDs) as ligands. The reaction of (NH4)5[Mo2Cl9]·H2O with the selected NSAIDs in methanol produced the complexes Mo2(μ-O2CR)4 where RCO2 is ibuprofen (1), naproxen (2), aspirin (3) and indomethacin (4). The products were obtained in good yields and extensively characterized with integrated techniques. Stability and solution behaviour were studied using a mixed experimental and computational approach. Finally, the biological activity of 1 and 3 (i.e. the most reactive and the most stable compounds of the series, respectively) was preliminarily assessed confirming the disassembling of the molecules in the biological milieu. Overall, some very interesting results emerged for these unconventional compounds from a mechanistic point of view.

The marine microalgae Ostreopsis cf. ovata are a well-known producer of palytoxin (PlTXs) analogues, i.e. ovatoxins (OVTXs) among others, which arouse concern for animal and human health. Both in field and laboratory studies, presence of OVTXs, detected in species directly feeding on O. cf. ovata, was frequently correlated with impairment on organisms' physiology, development and behaviour, while similar knowledge is still lacking for animals feeding on contaminated preys. In this study, transfer and toxicity of OVTXs were evaluated in an exposure experiment, in which gilthead seabream Sparus aurata was fed with bivalve mussel Mytilus galloprovincialis, contaminated by a toxic strain of O. cf. ovata. Mussels exposed to O. cf. ovata for 21 days accumulated meanly 188 ± 13 μg/kg OVTXs in the whole tissues. Seabreams fed with OVTX-contaminated mussels started to reject the food after 6 days of contaminated diet. Although no detectable levels of OVTXs were measured in muscle, liver, gills and gastro-intestinal tracts, the OVTX-enriched diet induced alterations of lipid metabolism in seabreams livers, displaying a decreased content of total lipid and fatty acid, together with overexpression of fatty acid biosynthetic genes, downregulation of β-oxidation genes and modulation of several genes related to lipid transport and regulation. Results from this study would suggest the hypothesis that OVTXs produced by O. cf. ovata may not be subject to bioaccumulation in fish fed on contaminated preys, being however responsible of significant biological effects, with important implications for human consumption of seafood products.

Raman MicroSpectroscopy (RMS) is a powerful label-free tool to probe the effects of drugs at a cellular/subcellular level. It is important, however, to be able to extract relevant biochemical and kinetic spectroscopic signatures of the specific cellular responses. In the present study, a combination of Multivariate Curve Resolution-Alternating Least Squares (MCR-ALS) and Principal Component Analysis (PCA) is used to analyse the RMS data for the example of exposure of primary Oral Squamous Carcinoma Cells (OSCC) to the chemotherapeutic agent cisplatin. Dosing regimens were established by cytotoxicity assays, and the effects of the drug on cellular spectral profiles were monitored from 16 to 72 hours post-exposure using an apoptosis assay, to establish the relative populations of viable (V), early (EA) and late apoptotic/dead (LA/D) cells after the drug treatment. Based on a kinetic model of the progression from V > EA > D, MCR-ALS regression analysis of the RMS responses was able to extract spectral profiles associated with each stage of the cellular responses, enabling a quantitative comparison of the response rates for the respective drug treatments. Moreover, PCA was used to compare the spectral profiles of the viable cells exposed to the drug. Spectral differences were highlighted in the early stages (16 hours exposure), indicative of the initial cellular response to the drug treatment, and also in the late stages (48-72 hours exposure), representing the cell death pathway. The study demonstrates that RMS coupled with multivariate analysis can be used to quantitatively monitor the progression of cellular responses to different drugs, towards future applications for label-free, in vitro, pre-clinical screening.