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Molecularly imprinted polymers (MIPs) rely on synthetic engineered materials able to selectively bind and intimately recognise a target molecule through its size and functionalities. The way in which MIPs interact with their targets, and the magnitude of this interaction, is closely linked to the chemical properties derived during the polymerisation stages, which tailor them to their specific target. Hence, MIPs are in-deep studied in terms of their sensitivity and cross-reactivity, further being used for monitoring purposes of analytes in complex analytical samples. As MIPs are involved in sensor development within different approaches, a systematic optimisation and rational data-driven sensing is fundamental to obtaining a best-performant MIP sensor. In addition, the closer integration of MIPs in sensor development requires that the inner properties of the materials in terms of sensitivity and selectivity are maintained in the presence of competitive molecules, which focus is currently opened. Identifying computational models capable of predicting and reporting the best-performant configuration of electrochemical sensors based on MIPs is of immense importance. The application of chemometrics using design of experiments (DoE) is nowadays increasingly adopted during optimisation problems, which largely reduce the number of experimental trials. These approaches, together with the emergent machine learning (ML) tool in sensor data processing, represent the future trend in design and management of point-of-care configurations based on MIP sensing. This review provides an overview on the recent application of chemometrics tools in optimisation problems during development and analytical assessment of electrochemical sensors based on MIP receptors. A comprehensive discussion is first presented to cover the recent advancements on response surface methodologies (RSM) in optimisation studies of MIPs design. Therefore, the recent advent of machine learning in sensor data processing will be focused on MIPs development and analytical detection in sensors.
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The present study employs X-ray photoelectron spectroscopy (XPS) to analyze plastic samples subjected to degradation processes with the aim to gain insight on the relevant chemical processes and disclose fragmentation mechanisms. Two model plastics, namely polystyrene (PS) and polyethylene (PE), are selected and analyzed before and after artificial UV radiation-triggered weathering, under simulated environmental hydrodynamic conditions, in fresh and marine water for different time intervals. The object of the study is to identify and quantify chemical groups possibly evidencing the occurrence of hydrolysis and oxidation reactions, which are the basis of degradation processes in the environment, determining macroplastic fragmentation. Artificially weathered plastic samples are analyzed also by Raman and FT-IR spectroscopy. Changes in surface chemistry with weathering are revealed by XPS, involving the increase in chemical moieties (hydroxyl, carbonyl, and carboxyl functionalities) which can be correlated with the degradation processes responsible for macroplastic fragmentation. On the other hand, the absence of significant modifications upon plastics weathering evidenced by Raman and FT-IR spectroscopy confirms the importance of investigating plastics surface, which represents the very first part of the materials exposed to degradation agents, thus revealing the power of XPS studies for this purpose. The XPS data on experimentally weathered particles are compared with ones obtained on microplastics collected from real marine environment for investigating the occurring degradation processes.
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Espectroscopía de Fotoelectrones , Plásticos , Polietileno , Espectroscopía de Fotoelectrones/métodos , Plásticos/química , Polietileno/química , Espectroscopía Infrarroja por Transformada de Fourier/métodos , Espectrometría Raman/métodos , Poliestirenos/química , Contaminantes Químicos del Agua/química , Contaminantes Químicos del Agua/análisis , Agua de Mar/química , Microplásticos/química , Oxidación-ReducciónRESUMEN
In the last two decades, fisheries and fish industries by-products have started to be recovered for the extraction of type I collagen because of issues related to the extraction of traditional mammalian tissues. In this work, special attention has been paid to by-products from fish bred in aquaponic plants. The valorization of aquaponic fish wastes as sources of biopolymers would make the derived materials eco-friendlier and attractive in terms of profitability and cost effectiveness. Among fish species, Nile Tilapia is the second-most farmed species in the world and its skin is commonly chosen as a collagen extraction source. However, to the best of our knowledge, no studies have been carried out to investigate, in depth, the age-related differences in fish skin with the final aim of selecting the most advantageous fish size for collagen extraction. In this work, the impact of age on the structural and compositional properties of Tilapia skin was evaluated with the aim of selecting the condition that best lends itself to the extraction of type I collagen for biomedical applications, based on the known fact that the properties of the original tissue have a significant impact on those of the final product. Performed analysis showed statistically significant age-related differences. In particular, an increase in skin thickness (+110 µm) and of wavy-like collagen fiber bundle diameter (+3 µm) besides their organization variation was observed with age. Additionally, a preferred collagen molecule orientation along two specific directions was revealed, with a higher fiber orientation degree according to age. Thermal analysis registered a shift of the endothermic peak (+1.7 °C) and an increase in the enthalpy (+3.3 J/g), while mechanical properties were found to be anisotropic, with an age-dependent brittle behavior. Water (+13%) and ash (+0.6%) contents were found to be directly proportional with age, as opposed to protein (-8%) and lipid (-10%) contents. The amino acid composition revealed a decrease in the valine, leucine, isoleucine, and threonine content and an increase in proline and hydroxyproline. Lastly, fatty acids C14:0, C15:0, C16:1, C18:2n6c, C18:3n6, C18:0, C20:3n3, and C23:0 were revealed to be upregulated, while C18:1n9c was downregulated with age.
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Cíclidos , Tilapia , Animales , Tilapia/metabolismo , Cíclidos/metabolismo , Colágeno Tipo I/metabolismo , Ácidos Grasos/metabolismo , Colágeno/metabolismo , MamíferosRESUMEN
One of the goals of archaeological studies is to determine how material goods and ideas moved among human populations, and bitumen is a worthy proxy because it has been used since prehistory. As a result, when bitumen is excavated from archaeological sites, determining its provenance is important because it sheds light on the trade and communication of populations at a given time. During the study of archaeological bitumen from coastal sites in central and southern Puglia (Italy), we observed that stable isotope ratios of saturated and aromatic fractions were incompatible with those obtained from asphaltenes, supporting the absorption of a foreign substance. Experiments showed that lipids are absorbed by bitumen and, in the case of oils, are distributed mainly in the saturated and aromatic fractions as their isotopic ratios change. The same experiments showed that the isotopic ratios of the asphaltenes do not change. Lipid absorption on the archaeological bitumen may have occurred before the bitumen was applied to the pottery, during the use of the pottery or while underground, before being excavated. These hypotheses are discussed, and it is concluded that the isotopic ratio of asphaltenes is a reliable proxy for provenance, whereas those of the saturated and aromatic fractions should be considered with caution due to possible lipid absorption. Nevertheless, they provide new information on pottery use that can be used in archaeological chemistry.
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Hidrocarburos , Aceites , Humanos , Hidrocarburos/química , Italia , IsótoposRESUMEN
Olive quick decline syndrome (OQDS) is a disorder associated with bacterial infections caused by Xylella fastidiosa subsp. pauca ST53 in olive trees. Metabolic profile changes occurring in infected olive trees are still poorly investigated, but have the potential to unravel reliable biomarkers to be exploited for early diagnosis of infections. In this study, an untargeted metabolomic method using high-performance liquid chromatography coupled to quadrupole-time-of-flight high-resolution mass spectrometry (HPLC-ESI-Q-TOF-MS) was used to detect differences in samples (leaves) from healthy (Ctrl) and infected (Xf) olive trees. Both unsupervised and supervised data analysis clearly differentiated the groups. Different metabolites have been identified as potential specific biomarkers, and their characterization strongly suggests that metabolism of flavonoids and long-chain fatty acids is perturbed in Xf samples. In particular, a decrease in the defence capabilities of the host after Xf infection is proposed because of a significant dysregulation of some metabolites belonging to flavonoid family. Moreover, oleic acid is confirmed as a putative diffusible signal factor (DSF). This study provides new insights into the host-pathogen interactions and confirms LC-HRMS-based metabolomics as a powerful approach for disease-associated biomarkers discovery in plants.
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Olea , Cromatografía Líquida de Alta Presión , Metabolómica , Enfermedades de las Plantas/microbiología , Espectrometría de Masas en TándemRESUMEN
PURPOSE: To explore the role of vacuum assisted closure (VAC) therapy versus conventional dressings in the Fournier's gangrene wound therapy. PATIENTS AND METHODS: This is a retrospective multi-institutional cohort study. Data of 92 patients from nine centers between 2007 and 2018 were retrospectively analyzed. After surgery, patient having a local or a disseminated FG were managed with VAC therapy or with conventional dressings. The 10-weeks wound closure cumulative rate and OS were analyzed. RESULTS: Of the 92 patients, 62 (67.4%) showed local and 30 (32.6%) a disseminated FG. After surgery, 19 patients (20.7%) with local and 14 (15.2%) with disseminated FG underwent to VAC therapy; 43 (46.7%) with local and 16 (17.4%) with disseminated FG were treated using conventional dressings. The multivariable logistic regression analysis demonstrated that the VAC in patients with disseminated FG led to a higher cumulative rate of wound closure than patients treated with no-VAC (OR = 6.5; 95% CI 1.1-37.4, p = 0.036). The Kaplan-Meier survival curves for the OS showed a significant difference between no-VAC patients with local and disseminated FG (OS rate at 90 days 0.90, 95% CI 0.71-0.97 vs 0.55, 95% CI 0.24-0.78, respectively; p = 0.039). Cox regression confirmed that no-VAC patients with disseminated FG showed the lowest OS (hazard ratio adjusted for sex and age HR = 3.4, 95% CI 1.1-10.4; p = 0.033). CONCLUSIONS: In this large cohort study, VAC therapy in patients with disseminated FG may offer an advantage in terms of 10-weeks wound closure cumulative rate and OS at 90 days after initial surgery.
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Vendajes , Gangrena de Fournier/cirugía , Terapia de Presión Negativa para Heridas , Anciano , Anciano de 80 o más Años , Estudios de Cohortes , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios RetrospectivosRESUMEN
Concentrated Growth Factors (CGF) represent new autologous (blood-derived biomaterial), attracting growing interest in the field of regenerative medicine. In this study, the chemical, structural, and biological characterization of CGF was carried out. CGF molecular characterization was performed by GC/MS to quantify small metabolites and by ELISA to measure growth factors and matrix metalloproteinases (MMPs) release; structural CGF characterization was carried out by SEM analysis and immunohistochemistry; CGF has been cultured, and its primary cells were isolated for the identification of their surface markers by flow cytometry, Western blot, and real-time PCR; finally, the osteogenic differentiation of CGF primary cells was evaluated through matrix mineralization by alizarin red staining and through mRNA quantification of osteogenic differentiation markers by real-time PCR. We found that CGF has a complex inner structure capable of influencing the release of growth factors, metabolites, and cells. These cells, which could regulate the production and release of the CGF growth factors, show stem features and are able to differentiate into osteoblasts producing a mineralized matrix. These data, taken together, highlight interesting new perspectives for the use of CGF in regenerative medicine.
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Diferenciación Celular , Péptidos y Proteínas de Señalización Intercelular/farmacología , Osteoblastos/citología , Osteogénesis , Células Madre/citología , Proliferación Celular , Células Cultivadas , Femenino , Humanos , Masculino , Osteoblastos/efectos de los fármacos , Osteoblastos/metabolismo , Células Madre/efectos de los fármacos , Células Madre/metabolismoRESUMEN
INTRODUCTION: Xylella fastidiosa (Xf) is a pathogenic bacterium that causes diseases in olive trees. Therefore, analytical methods for both the characterisation of the host/pathogen interaction and infection monitoring are needed. Volatile organic compounds (VOCs) are emitted by plants relate to their physiological state, therefore VOCs monitoring can assist in detecting stress or infection states before visible signs are present. OBJECTIVE: In this work, the headspace-solid phase microextraction-gaschromatography-mass spectrometry (HS-SPME-GC-MS) technique was used for the first time to highlight VOCs differences between healthy and Xf-infected olive trees. METHODOLOGY: VOCs from olive tree twig samples were extracted and analysed by HS-SPME-GC-MS, and hence identified by comparing the experimental linear retention indexes with the reference values and by MS data obtained from NIST library. Data were processed by principal component analysis (PCA) and analysis of variance (ANOVA). RESULTS: The HS-SPME step was optimised in terms of adsorbent phase and extraction time. HS-SPME-GC-MS technique was applied to the extraction and analysis of VOCs of healthy and Xf-infected olive trees. More than 100 compounds were identified and the differences between samples were evidenced by the multivariate analysis approach. The results showed the marked presence of methyl esters in Xf-infected samples, suggesting their probable involvement in the mechanism of diffusible signal factor. CONCLUSION: The proposed approach represents an easy and solvent-free method to evaluate the presence of Xf in olive trees, and to evidence volatiles produced by host/pathogen interactions that could be involved in the defensive mechanism of the olive tree and/or in the infective action of Xf.
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Cromatografía de Gases y Espectrometría de Masas/métodos , Olea/química , Olea/microbiología , Microextracción en Fase Sólida/métodos , Compuestos Orgánicos Volátiles/análisis , Xylella/patogenicidad , Análisis Multivariante , Olea/clasificaciónRESUMEN
Pirins are evolutionarily conserved iron-containing proteins that are found in all kingdoms of life, and have been implicated in diverse molecular processes, mostly associated with cellular stress. In the present study, we started from the evidence that the insertional inactivation of pirin-like gene SAM23877_RS18305 (pirA) by ΦC31 Att/Int system-based vectors in spiramycin-producing strain Streptomyces ambofaciens ATCC 23877 resulted in marked effects on central carbon and energy metabolism gene expression, high sensitivity to oxidative injury and repression of polyketide antibiotic production. By using integrated transcriptomic, proteomic and metabolite profiling, together with genetic complementation, we here show that most of these effects could be traced to the inability of the pirA-defective strain to modulate beta-oxidation pathway, leading to an unbalanced supply of precursor monomers for polyketide biosynthesis. Indeed, in silico protein-protein interaction modeling and in vitro experimental validation allowed us to demonstrate that PirA is a novel redox-sensitive negative modulator of very long-chain acyl-CoA dehydrogenase, which catalyzes the first committed step of the beta-oxidation pathway.
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Proteínas Bacterianas , Proteínas de Unión a Hierro , Ingeniería Metabólica , Streptomyces , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Proteínas de Unión a Hierro/genética , Proteínas de Unión a Hierro/metabolismo , Oxidación-Reducción , Policétidos/metabolismo , Streptomyces/genética , Streptomyces/metabolismoRESUMEN
INTRODUCTION: To report the long-term multicenter experience with retrograde intrarenal holmium-laser incision (RIR-HoLI) in the management of symptomatic renal sinus cysts (RSCs). In the literature, RIR-HoLI has been shown to be a safe and effective treatment, but there are only a few reports regarding long-term results and reproducibility of this procedure. MATERIAL AND METHODS: From June 2010 to June 2015, 14 patients with symptomatic RSCs underwent RIR-HoLI. The mean age was 52.1 ± 11.28 years (range 28-77) and the mean cyst size was 53.2 ± 14.23 mm (range 35-90). In all cases, contrast-enhanced computer tomography (CT) showed compression of the renal pelvis by the cyst (no malignancy). Surgical outcome was assessed in terms of symptoms improvement (measured by Visual Analogue Scale [VAS] for pain) and renal ultrasound findings at 3-6-12 months postoperatively and then yearly. CT scan was carried out at 12 months follow-up. RESULTS: RIR-HoLI was successful in all patients. The mean operative time was 47.8 ± 13.54 min (range 30-80) and mean hospital stay was 3.5 days (range 2-5). There were 2 Clavien grade II complications (flank pain and urgency delaying discharge). After surgery, all patients -became asymptomatic (VAS score change, p = 0.0001). One patient had persistence of a small cyst (10 mm). Mean follow-up is 44 ± 17.24 months (range 24-84); all patients remained asymptomatic, with no signs of recurrence. CONCLUSIONS: RIR-HoLI proved to be a safe and effective treatment for symptomatic RSCs. In our experience, it provided excellent long-term results and was reproducible at 4 different -institutions.
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Enfermedades Renales Quísticas/cirugía , Terapia por Láser/instrumentación , Láseres de Estado Sólido/uso terapéutico , Procedimientos Quirúrgicos Urológicos/instrumentación , Adulto , Anciano , Femenino , Humanos , Italia , Enfermedades Renales Quísticas/diagnóstico por imagen , Terapia por Láser/efectos adversos , Láseres de Estado Sólido/efectos adversos , Tiempo de Internación , Masculino , Persona de Mediana Edad , Tempo Operativo , Dimensión del Dolor , Complicaciones Posoperatorias/etiología , Factores de Tiempo , Tomografía Computarizada por Rayos X , Resultado del Tratamiento , Ultrasonografía , Ureteroscopía , Procedimientos Quirúrgicos Urológicos/efectos adversosRESUMEN
Heme is an essential molecule in many biological processes, such as transport and storage of oxygen and electron transfer as well as a structural component of hemoproteins. Defects of heme biosynthesis in developing erythroblasts have profound medical implications, as represented by sideroblastic anemia. The synthesis of heme requires the uptake of glycine into the mitochondrial matrix where glycine is condensed with succinyl coenzyme A to yield δ-aminolevulinic acid. Herein we describe the biochemical and molecular characterization of yeast Hem25p and human SLC25A38, providing evidence that they are mitochondrial carriers for glycine. In particular, the hem25Δ mutant manifests a defect in the biosynthesis of δ-aminolevulinic acid and displays reduced levels of downstream heme and mitochondrial cytochromes. The observed defects are rescued by complementation with yeast HEM25 or human SLC25A38 genes. Our results identify new proteins in the heme biosynthetic pathway and demonstrate that Hem25p and its human orthologue SLC25A38 are the main mitochondrial glycine transporters required for heme synthesis, providing definitive evidence of their previously proposed glycine transport function. Furthermore, our work may suggest new therapeutic approaches for the treatment of congenital sideroblastic anemia.
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Anemia/metabolismo , Hemo/biosíntesis , Mitocondrias/metabolismo , Proteínas de Transporte de Membrana Mitocondrial/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/metabolismo , Anemia/genética , Prueba de Complementación Genética , Glicina/genética , Glicina/metabolismo , Hemo/genética , Humanos , Mitocondrias/genética , Proteínas de Transporte de Membrana Mitocondrial/genética , Proteínas de Saccharomyces cerevisiae/genéticaRESUMEN
Rifamycins are mainstay agents in treatment of many widespread diseases, but how an improved rifamycin producer can be created is still incompletely understood. Here, we describe a comparative genomic approach to investigate the mutational patterns introduced by the classical mutate-and-screen method in the genome of an improved rifamycin producer. Comparing the genome of the rifamycin B overproducer Amycolatopsis mediterranei HP-130 with those of the reference strains A. mediterranei S699 and U32, we identified 250 variations, affecting 227 coding sequences (CDS), 109 of which were HP-130-specific since they were absent in both S699 and U32. Mutational and transcriptional patterns indicated a series of genomic manipulations that not only proved the causative effect of mutB2 (coding for methylmalonyl-CoA mutase large subunit) and argS2 (coding for arginyl tRNA synthetase) mutations on the overproduction of rifamycin, but also constituted a rational strategy to genetically engineer a reference strain into an overproducer.
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Actinobacteria/genética , Arginino-ARNt Ligasa/genética , Genoma Bacteriano/genética , Ingeniería Metabólica/métodos , Metilmalonil-CoA Mutasa/genética , Rifamicinas/metabolismo , Actinobacteria/clasificación , Mapeo Cromosómico/métodos , Hibridación Genómica Comparativa/métodos , Marcación de Gen/métodos , Mejoramiento Genético/métodos , Especificidad de la Especie , Regulación hacia Arriba/genéticaRESUMEN
In the setting of symptoms of testicular torsion the absence of diastolic flow or color flow on Doppler ultrasound has traditionally prompted emergent scrotal exploration. This practice emanates from the difficulty on ultrasound of distinguishing salvageable testes from those that are not salvageable. We evaluated the changes of echogenicity in the course of testicular torsion of the testis to identify characteristics predictive of irreversible organ damage.
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Torsión del Cordón Espermático/complicaciones , Torsión del Cordón Espermático/diagnóstico por imagen , Testículo/diagnóstico por imagen , Niño , Humanos , Masculino , Pronóstico , Estudios Retrospectivos , UltrasonografíaRESUMEN
Plastic pollution is a worldwide problem especially in the marine environment. Plastic items once fragmented into microplastics (MPs), can be captured by different marine species. Benthic filter feeders like sponges and polychaetas, due to their trophic strategy, are highly exposed to MPs pollution. Herein a simple but effective method to digest the fan worm Sabella spallanzanii and the calcareous sponge Paraleucilla magna is presented: a solution with KOH and H2O2 was able to remove quantitatively (more than 98 %) the organic matter in 3 h while an acid treatment dissolved most of spicules and chaetes in less than 30 min. MPs were easily identified both microscopically and spectroscopically on filters. Quantification in animals collected from the same environment showed that, on average, sponges accumulate fewer MPs than polychaetes (66 ± 31 and 117 ± 46 particles/g dry weight, respectively). The plastic recovery of the method was validated using three different approaches (spiking of standard PS microspheres, of common-use plastic objects, and of microplastics already weathered in marine environment). This procedure can make it easier and cost-effective to process biota in monitoring studies, providing information about bioindicator/bioremediation species.
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Microplastics (MPs) are dangerous and ubiquitous in the environment. The urgency to contrast plastic pollution is prompting the scientific community to offer new proposals. Recently, bioremediation using filter feeders is gaining consent as a nature-based solution. Herein four filter feeders (Mytilus galloprovincialis Lamarck, 1819, Sabella spallanzanii Gmelin, 1791, Phallusia mammillata Cuvier, 1815, Paraleucilla magna Klautau, Monteiro & Borojevic, 2004), studied in a previous laboratory experiment as MPs bioremediators, are evaluated in field conditions within 1-year. These organisms are part of an established fouling community growing on eco-friendly ropes in an Integrated Multi-Trophic Aquaculture (IMTA) in the Mar Grande of Taranto. After digesting the animal tissue, the MPs content was quantified by optical microscopy and spectroscopically characterized in the four seasons: highest values were measured in Autumn and lowest in Spring. M. galloprovincialis and P. mammillata were the most contaminated, but S. spallanzanii removed more MPs, due to its high density on the ropes. The whole community removed 3.15 × 107 MPs/season, with the amount of microfibers corresponding to a bottle cap/season.
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Herein, chemometric-assisted synthesis of electrochemical sensors based on electropolymerised ion-imprinted polymeric (e-IIP) films was explored. Co(II)-IIPs sensors were prepared by performing electropolymerisation procedures of polymerisation mixtures comprising varying concentrations of an electroactive o-aminophenol (o-AP) monomer and Co(II) ions, respectively, according to the Taguchi L9 experimental design, exploiting the simultaneous evaluation of other controlled parameters during electrosynthesis. Each e-IIP developed from Taguchi runs was compared with the respective non-imprinted polymer (NIP) films and fitted according to Langmuir-Freudlich isotherms. Distinctive patterns of low and high-affinity films were screened based on the qualities and properties of the developed IIPs in terms of binding kinetics (KD), imprinting factor, and the heterogeneity index of produced cavities. These results can provide a generic protocol for chemometric-assisted synthesis of e-IIPs based on poly-o-AP, providing highly stable, reproducible, and high-affinity imprinted polymeric films for monitoring purposes.
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Reconfiguration of chemical sensors, intended as the capacity of the sensor to adapt to novel operational scenarios, e.g., new target analytes, is potentially game changing and would enable rapid and cost-effective reaction to dynamic changes occurring at healthcare, environmental, and industrial levels. Yet, it is still a challenge, and rare examples of sensor reconfiguration have been reported to date. Here, we report on a reconfigurable label-free optical sensor leveraging the versatile immobilization of metal ions through a chelating agent on a nanostructured porous silica (PSiO2) optical transducer for the detection of different biomolecules. First, we show the reversible grafting of different metal ions on the PSiO2 surface, namely, Ni2+, Cu2+, Zn2+, and Fe3+, which can mediate the interaction with different biomolecules and be switched under mild conditions. Then, we demonstrate reconfiguration of the sensor at two levels: 1) switching of the metal ions on the PSiO2 surface from Cu2+ to Zn2+ and testing the ability of Cu2+-functionalized and Zn2+-reconfigured devices for the sensing of the dipeptide carnosine (CAR), leveraging the well-known chelating ability of CAR toward divalent metal ions; and 2) reconfiguration of the Cu2+-functionalized PSiO2 sensor for a different target analyte, namely, the nucleotide adenosine triphosphate (ATP), switching Cu2+ with Fe3+ ions to exploit the interaction with ATP through phosphate groups. The Cu2+-functionalized and Zn2+-reconfigured sensors show effective sensing performance in CAR detection, also evaluated in tissue samples from murine brain, and so does the Fe3+-reconfigured sensor toward ATP, thus demonstrating effective reconfiguration of the sensor with the proposed surface chemistry.
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Técnicas Biosensibles , Dióxido de Silicio , Animales , Ratones , Dióxido de Silicio/química , Técnicas Biosensibles/instrumentación , Técnicas Biosensibles/métodos , Nanoestructuras/química , Porosidad , Adenosina Trifosfato/análisis , Adenosina Trifosfato/química , Cobre/química , Metales/químicaRESUMEN
We report the case of a 46-year-old male hospitalized for abdominal pain and fever with history of a David procedure followed by an aortic valve replacement due to severe aortic regurgitation. Transesophageal echocardiography (TEE) and computed tomography showed a large mass floating in the aorta. After surgical excision of the vegetation, attached to the Dacron prosthesis, histological examination revealed Candida hyphae and spores confirming the diagnosis of a mycetoma in an ascending aorta tubular graft. At six-month follow-up, the patient was in good clinical condition without recurrence of the fungal mass on TEE.
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Aorta/diagnóstico por imagen , Aorta/microbiología , Implantación de Prótesis Vascular/métodos , Prótesis Vascular/microbiología , Candida/aislamiento & purificación , Micetoma/microbiología , Micetoma/cirugía , Infecciones Relacionadas con Prótesis/microbiología , Infecciones Relacionadas con Prótesis/cirugía , Válvula Aórtica/cirugía , Insuficiencia de la Válvula Aórtica/cirugía , Ecocardiografía Transesofágica , Estudios de Seguimiento , Implantación de Prótesis de Válvulas Cardíacas , Humanos , Masculino , Micetoma/diagnóstico por imagen , Infecciones Relacionadas con Prótesis/diagnóstico por imagen , Índice de Severidad de la Enfermedad , Resultado del TratamientoRESUMEN
OBJECTIVE: Recently, fine needle cytology (FNC) of the thyroid has been combined with biomolecular analysis. In particular, there has been detailed study of the V600E-BRAF mutation. The aim of our study is to demonstrate that with a single thyroid sample it is possible to obtain enough cellular material for both cytological diagnosis and a V600E-BRAF molecular test. STUDY DESIGN: FNC was carried out under ultrasound guidance aided by an echographist and cytopathologist. We acquired one biopsy for each nodule with a 23-gauge needle without suction. The preparations were smeared by the pathologist onto one glass slide, air dried and stained with Diff-Quick. Cell adequacy was evaluated for each patient. The needle was washed by aspirating 2 ml of physiologic solution which was collected into a tube. The material was collected for molecular testing. RESULTS: The following cytological diagnoses were made: not neoplastic, Tir2 (n = 227); indeterminate, Tir3 (n = 15); suspicious, Tir4 (n = 4), and malignancy, Tir5 (n = 12). The V600E-BRAF mutation was found in 0 of 227 Tir2 specimens, 2 of 15 (13.3%) Tir3 specimens, 2 of 4 (50%) Tir4 specimens and 9 of 12 (75%) Tir5 specimens. CONCLUSIONS: Our data showed that, in a routine clinical setting, FNC specimens can be handled properly to provide both morphological and molecular information. In fact, our tests show that with a single specimen it is possible to set up a slide for the cytological diagnosis and to obtain enough residual cellular material for DNA extraction (>70 ng) and for the identification of the V600E-BRAF mutation.
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Biopsia con Aguja Fina , Citodiagnóstico/métodos , Proteínas Proto-Oncogénicas B-raf/genética , Nódulo Tiroideo/genética , Nódulo Tiroideo/patología , Humanos , Reacción en Cadena de la Polimerasa , Proteínas Proto-Oncogénicas B-raf/análisis , Sensibilidad y EspecificidadRESUMEN
Due to the increased resistance to all available antibiotics and the lack of vaccines, Neisseria gonorrhoeae (the gonococcus) poses an urgent threat. Although the mechanisms of virulence and antibiotic resistance have been largely investigated in this bacterium, very few studies have addressed the stringent response (SR) that in pathogenic bacteria controls the expression of genes involved in host-pathogen interaction and tolerance and persistence toward antibiotics. In this study, the results of the transcriptome analysis of a clinical isolate of N. gonorrhoeae, after induction of the SR by serine hydroxamate, provided us with an accurate list of genes that are transcriptionally modulated during the SR. The list includes genes associated with metabolism, cellular machine functions, host-pathogen interaction, genome plasticity, and antibiotic tolerance and persistence. Moreover, we found that the artificial induction of the SR in N. gonorrhoeae by serine hydroxamate is prevented by thiostrepton, a thiopeptide antibiotic that is known to interact with ribosomal protein L11, thereby inhibiting functions of EF-Tu and EF-G, and binding of pppGpp synthase I (RelA) to ribosome upon entry of uncharged tRNA. We found that N. gonorrhoeae is highly sensitive to thiostrepton under in vitro conditions, and that thiostrepton, in contrast to other antibiotics, does not induce tolerance or persistence. Finally, we observed that thiostrepton attenuated the expression of key genes involved in the host-pathogen interaction. These properties make thiostrepton a good drug candidate for dampening bacterial virulence and preventing antibiotic tolerance and persistence. The ongoing challenge is to increase the bioavailability of thiostrepton through the use of chemistry and nanotechnology.