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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, 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.
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Quadruplex G , Guanosina , Hidrogéis , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Guanosina/química , Hidrogéis/química , Potássio/química , Potássio/análise , Vibração , Guanosina Monofosfato/químicaRESUMO
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.
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Anti-Inflamatórios não Esteroides , Complexos de Coordenação , Molibdênio , Anti-Inflamatórios não Esteroides/química , Molibdênio/química , Complexos de Coordenação/química , Ligantes , Humanos , Naproxeno/química , Animais , Ibuprofeno/química , Aspirina/química , Indometacina/química , CamundongosRESUMO
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 µm, 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 ~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.
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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.
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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.
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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.
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Sistema Digestório , Mytilus , Poluentes Químicos da Água , Animais , Mytilus/efeitos dos fármacos , Mytilus/metabolismo , Poluentes Químicos da Água/toxicidade , Sistema Digestório/efeitos dos fármacos , Sistema Digestório/metabolismo , Substâncias Macromoleculares , Carbamazepina/farmacologia , Espectroscopia de Infravermelho com Transformada de Fourier , Bivalves/efeitos dos fármacos , Bivalves/químicaRESUMO
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 ß-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.
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Dinoflagellida , Mytilus , Dourada , Animais , Humanos , Toxinas Marinhas/toxicidade , Metabolismo dos Lipídeos , Alimentos Marinhos , Dinoflagellida/genética , Ácidos Graxos , LipídeosRESUMO
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 .
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Implantes Dentários , Colágeno , Gengiva/patologia , Lasers , Osseointegração , Propriedades de Superfície , HumanosRESUMO
Lyotropic Liquid Crystalline (LLC) nanoparticles represent an emerging class of smart, biocompatible, and biodegradable systems for the delivery of drugs. Among these, structures with complex 3D architectures such as cubosomes are of particular interest. These are non- lamellar assemblies having hydrophobic and hydrophilic portions able to carry drugs of different nature. They can further be modulated including suitable additives to control the release of the active payload, and to promote an active targeting. Starting from monoolein (GMO) cubic phase, different concentrations of mannose-based esters were added, and the eventual structural modifications were monitored to ascertain the effects of the presence of glycolipids. Moreover, the structural properties of these nanosystems loaded with Dexamethasone (DEX), a very well-known anti-inflammatory steroid, were also studied. Experiments were carried out by synchrotron Small Angle X-ray Scattering (SAXS), Raman Microspectroscopy (RMS) and Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR) measurements. The drug delivery potential (i.e. entrapment efficiency and release properties) of the obtained nanoparticles was evaluated. Finally, in vitro cytocompatibility and anti-inflammatory activity studies of the prepared formulations were carried out. Inclusion of mannose-based surfactants up to 10 mol% influenced the structural parameters of Im3m cubic phase and swollen cubic phases were obtained with the different glycolipids with lattice parameters significantly higher than GMO. A complete cytocompatibility and an increased DEX activity were observed, thus suggesting the possibility to use GMO/glycolipids nanoparticles to formulate innovative drug delivery systems.
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Cristais Líquidos , Manose , Espalhamento a Baixo Ângulo , Difração de Raios X , Sistemas de Liberação de Medicamentos , Anti-Inflamatórios/farmacologia , Glicolipídeos , Cristais Líquidos/químicaRESUMO
OBJECTIVES: This in vitro study aimed assessing the remineralization potential of three commercial fluoride-based toothpastes in permanent teeth with natural white spot lesions (WSLs). A multidisciplinary approach based on Raman microspectroscopy (RMS), Scanning electron microscopy (SEM), Energy-dispersive x-ray spectroscopy (EDS), and Vickers microhardness (VMH) was exploited. METHODS: N = 12 human molars with natural WSLs in the proximal-vestibular zone were selected and divided into 4 groups (n = 3) according to the different treatments: HAF (hydroxyapatite with fluoride ions); SMF (sodium monofluorophosphate with arginine); SF (sodium fluoride with enzymes), and CTRL (untreated group). All toothpastes tested contained 1450 ppm of fluoride. Teeth samples were submitted to the following protocol: a 7-day pH cycling treatment, with two daily exposures (2 min each time) to the commercial toothpastes described above. The surface micromorphology (SEM), the chemical/elemental composition (RMS and EDS), and the Vickers microhardness (VMH) were evaluated. Statistical analysis was performed. RESULTS: A remarkable remineralization of WSLs in SEM images was observed in all treated groups compared to CTRL. In particular, HAF and SF displayed higher values of VMH, phosphates amount (I960), crystallinity (FWHM960), and lower ones of C/P (I1070/I960) with respect to CTRL. Intermediate values were found in SMF, higher than CTRL but lower with respect to HAF and SF. As regards the Ca/P ratio, statistically significant differences (p < 0.05) were found between SF and the other groups. CONCLUSIONS: All the tested dentifrices have shown to remineralize the WSLs. SF and HAF have comparable capability in hardness recovery and crystallinity; however, SF shows the best remineralizing potential according to both micromorphological and chemical analyses. Clinical relevance The daily use of toothpastes containing hydroxyapatite partially replaced with fluoride, sodium monofluorophosphate with arginine and sodium fluoride toothpaste associated with enzymes represents a preventive, therapeutic, effective, and non-invasive tool for remineralize WSLs.
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Cárie Dentária , Fluoretos , Humanos , Fluoretos/farmacologia , Cremes Dentais/farmacologia , Cremes Dentais/uso terapêutico , Fluoreto de Sódio/farmacologia , Fluoreto de Sódio/uso terapêutico , Esmalte Dentário , Cárie Dentária/tratamento farmacológico , Cárie Dentária/prevenção & controle , Arginina/farmacologia , Hidroxiapatitas/farmacologia , Remineralização Dentária/métodos , Cariostáticos/uso terapêuticoRESUMO
The potential toxicity of microplastics is a growing concern for the scientific community. The loggerhead sea turtle (Caretta caretta) is particularly inclined to accidently ingest plastic and microplastic due to its long-life cycle features. The possible transfer of microplastics from the female to the eggs should be investigated. The present study investigated the presence of microplastics in yolk and liver samples evaluating the number of melanomacrophages in the hepatic tissue as a possible biomarker of microplastics impact on the embryonic health status. The biometric parameters and liver histological analysis of 27 and 48 embryos (from two different nests respectively) at the 30 stage of development were analyzed. Raman Microspectroscopy was performed to identify the microplastics after alkaline digestion (10% KOH) of yolk and portion of liver from 5 embryos at the 30 developmental stage per nest. Microplastics were found in yolk and liver of loggerhead sea turtles at late embryonic stage for the first time. All microplastics were smaller than 5 µm and were made of polymers and colors suggesting their diverse origins. A total number of 21 microplastics, with dimensions lower than 5 µm, were found between the two nests (11 and 10 microplastics respectively). Only two shape categories were identified: spheres and fragments. The most frequent polymers observed were polyethylene, polyvinyl chloride and acrylonitrile butadiene styrene (31.5%, 21.1% and 15.8% respectively). Despite the eggs showing a higher number of microplastics in yolk samples than liver (15 and 6 microplastics in yolk and liver respectively), a positive correlation was observed only between the number of melanomacrophages (r = 0.863 p < 0.001) and microplastics in the liver. This result may suggest that microplastics could exert some effects on the hepatic tissues. Future studies should investigate this aspect and the possible relation between microplastics and other stress biomarkers.
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Microplásticos , Tartarugas , Animais , Feminino , Plásticos , Projetos Piloto , FígadoRESUMO
The presence of microplastics (MPs) in human fluids and organs is a great concern, since, as highlighted by recent studies on animal models, they could cause alterations of several physiological functions, including reproduction. In this study, semen samples collected from men living in a polluted area of the Campania Region (Southern Italy), were analyzed to assess the presence of MPs. N. 16 pigmented microplastic fragments (ranging from 2 to 6 µm in size) with spheric or irregular shapes were found in six out of ten samples. All the detected MPs were characterized in terms of morphology (size, colour, and shape) and chemical composition by Raman Microspectroscopy. Chemical composition showed the presence of polypropylene (PP), polyethylene (PE), polyethylene terephthalate (PET), polystyrene (PS), polyvinylchloride (PVC), polycarbonate (PC), polyoxymethylene (POM) and acrylic, suggesting ingestion and/or inhalation as a route of exposure to environmental MPs. In this work, we propose for the first time a mechanism by which MPs pass into the semen most likely through the epididymis and seminal vesicles, which are the most susceptible to inflammation.
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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.
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Carcinoma de Células Escamosas , Cisplatino , Humanos , Cisplatino/farmacologia , Análise dos Mínimos Quadrados , Análise Espectral Raman/métodos , Carcinoma de Células Escamosas/tratamento farmacológico , Análise MultivariadaRESUMO
Gestational diabetes mellitus (GDM) and small-for-gestational-age (SGA) are two metabolic-related diseases that could affect women during pregnancy. Considering that the chorionic villi (CVs) are crucial structures for the feto-maternal exchange, the alterations in their conformation have been linked to an imbalanced metabolic environment of placenta. In this study, a multidisciplinary approach has been carried out to describe the changes occurring in the placental CVs of GDM and SGA patients. The results revealed higher levels of superoxide dismutase 1 (SOD-1) and catalase (CAT), especially in the GDM placentae, which could be correlated with the hyperglycemic environment characteristic of this pathology. Furthermore, spectroscopy and histologic analyses revealed that both pathologies modify the placental lipid composition altering its structure. However, SGA induces lipid peroxidation and reduces collagen deposition within the CVs. Since the endocannabinoid system (ECS) is involved in placentation and different metabolic activities, the cannabinoid receptor 1 (CB1) and transient receptor potential cation channel subfamily V member 1 (TRPV-1) were analyzed. No changes have been observed either at general or specific levels in the CVs comparing control and pathological samples, suggesting the non-involvement of the cannabinoid system in these two pathologies.
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Diabetes Gestacional , Humanos , Recém-Nascido , Gravidez , Feminino , Diabetes Gestacional/metabolismo , Placenta/metabolismo , Placentação , Recém-Nascido Pequeno para a Idade Gestacional , Biomarcadores/metabolismoRESUMO
A nanodelivery system based on palladium nanoparticles (PdNP) and cisplatin (CisPt) was developed by physisorption of the drug onto the PdNP synthesized via a green redox process, using d-glucose and polyvinylpyrrolidone (PVP) as reducing and stabilizing/capping agents, respectively. UV-vis analysis and H2-evolution measurements were carried out to prove the nanoparticles' capability to act as bimodal theranostic nanomedicine, i.e., having both plasmonic and photocatalytic properties. XPS, XRD, and TEM allowed light to be shed on the chemical composition and morphology of the PdNP. The analysis of the UV-visible spectra evidenced plasmonic peak changes for the hybrid nanoparticle-drug assembly (Pd@CisPt), which pointed to a significant interaction of CisPt with the NP surface. The drug loading was quantitatively estimated by ICP-OES measurements, while DLS and AFM confirmed the strong association of the drug with the nanoparticle surface. The test of SOD-like activity in a cell-free environment proved the maintenance of the antioxidant capability of PdNP also in the Pd@CisPt systems. Finally, Pd@CisPt tested in prostate cancer cells (PC-3 line) unveiled the antitumoral action of the developed nanomedicine, related to reactive oxygen species (ROS) generation, with a condition of protein misfolding/unfolding and DNA damage, as evidenced by cytotoxicity and MitoSOX assays, as well as Raman microspectroscopy, respectively. Cell imaging by confocal microscopy evidenced cellular uptake of the nanoparticles, as well as dynamic processes of copper ion accumulation at the level of subcellular compartments. Finally, cell migration studies upon treatment with Pd@CisPt evidenced a tunable response between the inhibitory effect of CisPt and the enhanced rate of cell migration for the metal NP alone, which pointed out the promising potential of the developed theranostic nanomedicine in tissue regeneration.
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Antineoplásicos , Nanopartículas Metálicas , Masculino , Humanos , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Antineoplásicos/química , Nanopartículas Metálicas/uso terapêutico , Nanopartículas Metálicas/química , Nanomedicina Teranóstica/métodos , Paládio/farmacologia , Paládio/química , Cisplatino/químicaRESUMO
Clear orthodontic aligners have recently seen increasing popularity. The thermoplastic materials present several advantages, even if it is known that all plastic products can be subjected to environmental and mechanical degradation, leading to the release of microplastics (MPs). Their ingestion could cause oxidative stress and inflammatory lesions. This study aims to evaluate the potential detachment of MPs by clear aligners due to mechanical friction simulated with a 7-day protocol in artificial saliva. The study was performed on orthodontic clear aligners from different manufacturers: Alleo (AL); FlexiLigner (FL); F22 Aligner (F22); Invisalign® (INV); Lineo (LIN); Arc Angel (ARC), and Ortobel Aligner (OR). For each group, two aligners were immersed in artificial saliva for 7 days and stirred for 5 h/day, simulating the physiological teeth mechanical friction. After 7 days, the artificial saliva was filtered; then, filters were analyzed by Raman Microspectroscopy (RMS) and Scanning Electron Microscopy (SEM), respectively to chemically identify the polymeric matrix and to measure the number and size of the detected MPs. RMS spectra revealed that AL, FL, LIN, ARC, and OR aligners were composed by polyethylene terephthalate, while F22 and INV ones by polyurethane. SEM analysis showed that the highest number of MPs was found in ARC and the lowest in INV (p < 0.05). As regards MPs' size, no statistically significant difference was found among groups, with most MPs ranging from 5 to 20 µm. Noteworthy, a highly significant correlation (p < 0.0001) was highlighted between the distribution of MPs size and the different typologies of aligners. This in vitro study highlighted for the first time the detachment of MPs from clear aligners due to mechanical friction. This evidence may represent a great concern in the clinical practice since it could impact human general health.
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Aparelhos Ortodônticos Removíveis , Plásticos , Humanos , Saliva Artificial , Microplásticos , Análise EspectralRESUMO
Reliable point-of-care (POC) rapid tests are crucial to detect infection and contain the spread of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). The emergence of several variants of concern (VOC) can reduce binding affinity to diagnostic antibodies, limiting the efficacy of the currently adopted tests, while showing unaltered or increased affinity for the host receptor, angiotensin converting enzyme 2 (ACE2). We present a graphene field-effect transistor (gFET) biosensor design, which exploits the Spike-ACE2 interaction, the crucial step for SARS-CoV-2 infection. Extensive computational analyses show that a chimeric ACE2-Fragment crystallizable (ACE2-Fc) construct mimics the native receptor dimeric conformation. ACE2-Fc functionalized gFET allows in vitro detection of the trimeric Spike protein, outperforming functionalization with a diagnostic antibody or with the soluble ACE2 portion, resulting in a sensitivity of 20 pg/mL. Our miniaturized POC biosensor successfully detects B.1.610 (pre-VOC), Alpha, Beta, Gamma, Delta, Omicron (i.e., BA.1, BA.2, BA.4, BA.5, BA.2.75 and BQ.1) variants in isolated viruses and patient's clinical nasopharyngeal swabs. The biosensor reached a Limit Of Detection (LOD) of 65 cps/mL in swab specimens of Omicron BA.5. Our approach paves the way for a new and reusable class of highly sensitive, rapid and variant-robust SARS-CoV-2 detection systems.
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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.
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Preeclampsia is a human pregnancy-specific disease characterized by abnormal placentation that usually presents with maternal hypertension and proteinuria. The main hallmark of preeclampsia, impaired trophoblast migration, and the subsequent disruption of uterine arteries remodeling lead to several molecular alterations in the placental compartments with those occurring in the chorionic villi being of the utmost importance. Given the essential role of the endocannabinoid system during preimplantation and trophoblast migration, we have combined the histological and hyperspectral imaging analyses to shed light on the involvement of two cannabinoid receptors in the macromolecular alterations related to preeclampsia. The results obtained by immunohistochemistry showed a significant increase in the protein levels of cannabinoid receptor 1 (CB1) in the preeclamptic chorionic villi. However, no changes were reported regarding transient receptor potential vanilloid 1 (TRPV-1) levels either in the bulk placental samples or chorionic villi when comparing control and preeclamptic patients. Histological analysis and Fourier-transform infrared spectroscopy (FTIRI) showed an increase in collagen deposition together with higher levels of lipid peroxidation and phosphorylated compounds in the pathological villi. Since CB1 enhancement has been described as promoting fibrosis and oxidative stress in several tissues, we proposed that the higher receptor abundance in preeclampsia could be triggering similar molecular effects in preeclamptic term placentas.
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Vilosidades Coriônicas , Pré-Eclâmpsia , Humanos , Feminino , Gravidez , Vilosidades Coriônicas/metabolismo , Placenta/metabolismo , Pré-Eclâmpsia/metabolismo , Placentação , Trofoblastos/metabolismo , Receptores de Canabinoides/metabolismoRESUMO
Microplastics (MPs) are defined as plastic particles smaller than 5 mm. They have been found almost everywhere they have been searched for and recent discoveries have also demonstrated their presence in human placenta, blood, meconium, and breastmilk, but their location and toxicity to humans have not been reported to date. The aim of this study was twofold: 1. To locate MPs within the intra/extracellular compartment in human placenta. 2. To understand whether their presence and location are associated with possible structural changes of cell organelles. Using variable pressure scanning electron microscopy and transmission electron microscopy, MPs have been localized in ten human placentas. In this study, we demonstrated for the first time the presence and localization in the cellular compartment of fragments compatible with MPs in the human placenta and we hypothesized a possible correlation between their presence and important ultrastructural alterations of some intracytoplasmic organelles (mitochondria and endoplasmic reticulum). These alterations have never been reported in normal healthy term pregnancies until today. They could be the result of a prolonged attempt to remove and destroy the plastic particles inside the placental tissue. The presence of virtually indestructible particles in term human placenta could contribute to the activation of pathological traits, such as oxidative stress, apoptosis, and inflammation, characteristic of metabolic disorders underlying obesity, diabetes, and metabolic syndrome and partially accounting for the recent epidemic of non-communicable diseases.