Fluorescence Sensing Using DNA Aptamers in Cancer Research and Clinical Diagnostics

Among the various advantages of aptamers over antibodies, remarkable is their ability to tolerate a large number of chemical modifications within their backbone or at the termini without losing significant activity. Indeed, aptamers can be easily equipped with a wide variety of reporter groups or coupled to different carriers, nanoparticles, or other biomolecules, thus producing valuable molecular recognition tools effective for diagnostic and therapeutic purposes. This review reports an updated overview on fluorescent DNA aptamers, designed to recognize significant cancer biomarkers both in soluble or membrane-bound form. In many examples, the aptamer secondary structure switches induced by target recognition are suitably translated in a detectable fluorescent signal using either fluorescently-labelled or label-free aptamers. The fluorescence emission changes, producing an enhancement ("signal-on") or a quenching ("signal-off") effect, directly reflect the extent of the binding, thereby allowing for quantitative determination of the target in bioanalytical assays. Furthermore, several aptamers conjugated to fluorescent probes proved to be effective for applications in tumour diagnosis and intraoperative surgery, producing tumour-type specific, non-invasive in vivo imaging tools for cancer pre- and post-treatment assessment

Toxic organic contaminants in airborne particles: levels, potential sources and risk assessment

In the last years, many studies have focused on risk assessment of exposure of workers to airborne particulate matter (PM). Several studies indicate a strong correlation between PM and adverse health outcomes, as a function of particle size. In the last years, the study of atmospheric particulate matter has focused more on particles less than 10 m or 2.5 m in diameter; however, recent studies identify in particles less than 0.1 m the main responsibility for negative cardiovascular effects. The present paper deals with the determination of 66 organic compounds belonging to six different classes of persistent organic pollutants (POPs) in the ultrafine, fine and coarse fractions of PM (PM < 0.1 m; 0.1 < PM < 2.5 mand 2.5 < PM < 10 m) collected in three outdoor workplaces and in an urban outdoor area. Data obtained were analyzed with principal component analysis (PCA), in order to underline possible correlation between sites and classes of pollutants and characteristic emission sources. Emission source studies are, in fact, a valuable tool for both identifying the type of emission source and estimating the strength of each contamination source, as useful indicator of environment healthiness. Moreover, both carcinogenic and non-carcinogenic risks were determined in order to estimate human health risk associated to study sites. Risk analysis was carried out evaluating the contribution of pollutant distribution in PM size fractions for all the sites. The results highlighted significant differences between the sites and specific sources of pollutants related to work activities were identified. In all the sites and for all the size fractions of PM both carcinogenic and non-carcinogenic risk values were below acceptable and safe levels of risks recommended by the regulatory agencies

Gut microbiota profile in infants with milk and/or egg allergy and evaluation of intestinal colonization and persistence of a probiotic mixture

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Enhanced expression of hepatic lipogenic enzymes in an animal model of sedentariness.

The hindlimb-suspended rat was used as animal model to investigate the effects induced by immobilization of the skeletal muscle in the expression of the genes encoding hepatic lipogenic enzymes. Following a 14-day period of immobilization, rats were injected intraperitoneally with radioactive acetate, and the labeling of hepatic lipids and cholesterol was evaluated 15 min after the isotope injection. The incorporation of labeled acetate in lipids and cholesterol was almost three times higher in the liver of immobilized rats than in control animals as a consequence of the enhanced transcription of the genes encoding acetyl-CoA synthase, acetyl-CoA carboxylase, fatty acid synthase, and 3-hydroxy-3-methylglutaryl-CoA reductase. The high expression of the key enzymes for fatty acid and cholesterol synthesis induced by immobilization was not paralleled by an increase of the hepatic sterol-regulatory element binding protein (SREBP)-1 and SREBP-2 mRNA content. However, the expression of the mature form of SREBP-1 and SREBP-2 was higher in the nuclear fraction of immobilized rat liver than in controls due to a significant increase of the cleavage of the native proteins. Immobilization also affected the expression of proteins involved in lipid degradation. In fact, the hepatic content of peroxisome proliferator-activated receptor-alpha (PPARalpha) mRNA and of PPARalpha target genes encoding carnitine palmitoyl transferase-1 and acyl-CoA oxidase were significantly increased upon immobilization

Testing e validazione comparativa di coating idrofobizzanti per superfici in gelcoat

Nell’industria delle telecomunicazioni a supporto del settore aerospaziale, il mantenimento dell’efficacia e della sicurezza delle trasmissioni in condizioni ambientali estreme assume un’importanza crescente, in funzione della sempre maggiore diffusione dei siti trasmittenti/riceventi, operativi su scala planetaria. In questa cornice, si richiede che gli elementi a protezione dei dispositivi installati si dimostrino durevoli nel tempo anche quando esposti ad abbondanti precipitazioni, accumuli nevosi continuati o stratificazioni di ghiaccio, e improvvisi shock termici, garantendo nel contempo la fedeltà della trasmissione. Una marcata idrofobicità degli strati esterni delle coperture risulta quindi alla base per mantenerne l’efficacia nel tempo. Nel presente studio sono vengono riportati e confrontati i risultati di una serie di trattamenti idrofobizzanti su superfici di elementi tecnici in gelcoat costruiti per strutture radome, insieme a prove di resistenza meccanica, al fine di simulare in laboratorio le effettive condizioni di esposizione al danneggiamento. A questo scopo, le superfici sono state sottoposte a prove di bagnabilità tramite misurazione dell’ angolo di contatto e a test di resistenza meccanica alla scalfittura e di aderenza dei rivestimenti applicati, tramite Pencil Hardness Test, Cross-cut test, Scotch tape Test e pull-off Test. I campioni sono stati sottoposti ad un processo di invecchiamento accelerato mirato, alfine di valutare il mantenimento delle proprietà nel tempo. La comparazione multiparametrica dei risultati ottenuti ha reso possibile individuare i trattamenti che esibiscono performance eccellenti in termini di proprietà ultra-idrofobizzanti coniugate alla resistenza meccanica, anche mantenute nel tempo

Polychlorinated biphenyl profile in polyhydroxy-alkanoates synthetized from urban organicwastes

Call CIRC-05-2016The microbial synthesis of polyhydroxyalkanoates (PHA) from organic wastes is a valuable process to valorize available renewable resources, such as food wastes and biological sludge. Bioplastics find many applications in various sectors, from medical field to food industry. However, persistent organic pollutants could be transferred from wastes to the final product. The present paper demonstrates that the use of municipal wastes in PHA production is safe for the environment and human health and provides a polychlorinated biphenyl (PCB) profile in both commercial and waste-based PHA samples. PCB analysis in several PHA samples showed very low concentrations of the target analytes. Commercial PHA samples showed a similar PCB level with respect to PHA samples from municipal waste/sludge and higher than PHA samples from fruit waste. For all analyzed PCBs, detected concentrations were consistently lower than the ones reported in regulatory framework or guidelines.publishersversionpublishe

Fighting the Huntington's Disease with a G-Quadruplex-Forming Aptamer Specifically Binding to Mutant Huntingtin Protein: Biophysical Characterization, In Vitro and In Vivo Studies

A set of guanine-rich aptamers able to preferentially recognize full-length huntingtin with an expanded polyglutamine tract has been recently identified, showing high efficacy in modulating the functions of the mutated protein in a variety of cell experiments. We here report a detailed biophysical characterization of the best aptamer in the series, named MS3, proved to adopt a stable, parallel G-quadruplex structure and show high nuclease resistance in serum. Confocal microscopy experiments on HeLa and SH-SY5Y cells, as models of non-neuronal and neuronal cells, respectively, showed a rapid, dose-dependent uptake of fluorescein-labelled MS3, demonstrating its effective internalization, even in the absence of transfecting agents, with no general cytotoxicity. Then, using a well-established Drosophila melanogaster model for Huntington's disease, which expresses the mutated form of human huntingtin, a significant improvement in the motor neuronal function in flies fed with MS3 was observed, proving the in vivo efficacy of this aptamer

Truncated Analogues of a G-Quadruplex-Forming Aptamer Targeting Mutant Huntingtin: Shorter Is Better!

Two analogues of the MS3 aptamer, which was previously shown to have an exquisite capability to selectively bind and modulate the activity of mutant huntingtin (mHTT), have been here designed and evaluated in their physicochemical and biological properties. Featured by a distinctive propensity to form complex G-quadruplex structures, including large multimeric aggregates, the original 36-mer MS3 has been truncated to give a 33-mer (here named MS3-33) and a 17-mer (here named MS3-17). A combined use of different techniques (UV, CD, DSC, gel electrophoresis) allowed a detailed physicochemical characterization of these novel G-quadruplex-forming aptamers, tested in vitro on SH-SY5Y cells and in vivo on a Drosophila Huntington’s disease model, in which these shorter MS3-derived oligonucleotides proved to have improved bioactivity in comparison with the parent aptamer