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1.
Biosens Bioelectron ; 262: 116544, 2024 Jun 29.
Artículo en Inglés | MEDLINE | ID: mdl-38963952

RESUMEN

In this work, a nanostructured conductive film possessing nanozyme features was straightforwardly produced via laser-assembling and integrated into complete nitrocellulose sensors; the cellulosic substrate allows to host live cells, while the nanostructured film nanozyme activity ensures the enzyme-free real-time detection of hydrogen peroxide (H2O2) released by the sames. In detail, a highly exfoliated reduced graphene oxide 3D film decorated with naked platinum nanocubes was produced using a CO2-laser plotter via the simultaneous reduction and patterning of graphene oxide and platinum cations; the nanostructured film was integrated into a nitrocellulose substrate and the complete sensor was manufactured using an affordable semi-automatic printing approach. The linear range for the direct H2O2 determination was 0.5-80 µM (R2 = 0.9943), with a limit of detection of 0.2 µM. Live cell measurements were achieved by placing the sensor in the culture medium, ensuring their adhesion on the sensors' surface; two cell lines were used as non-tumorigenic (Vero cells) and tumorigenic (SKBR3 cells) models, respectively. Real-time detection of H2O2 released by cells upon stimulation with phorbol ester was carried out; the nitrocellulose sensor returned on-site and real-time quantitative information on the H2O2 released proving useful sensitivity and selectivity, allowing to distinguish tumorigenic cells. The proposed strategy allows low-cost in-series semi-automatic production of paper-based point-of-care devices using simple benchtop instrumentation, paving the way for the easy and affordable monitoring of the cytopathology state of cancer cells.

2.
Biochim Biophys Acta Mol Cell Biol Lipids ; 1869(7): 159524, 2024 Jun 08.
Artículo en Inglés | MEDLINE | ID: mdl-38857757

RESUMEN

Neuroinflammation is a hallmark of several neurodegenerative disorders that has been extensively studied in recent years. Microglia, the primary immune cells of the central nervous system (CNS), are key players in this physiological process, demonstrating a remarkable adaptability in responding to various stimuli in the eye and the brain. Within the complex network of neuroinflammatory signals, the fatty acid N-ethanolamines, in particular N-arachidonylethanolamine (anandamide, AEA), emerged as crucial regulators of microglial activity under both physiological and pathological states. In this study, we interrogated for the first time the impact of the signaling of these bioactive lipids on microglial cell responses to a sub-lethal acute UVB radiation, a physical stressor responsible of microglia reactivity in either the retina or the brain. To this end, we developed an in vitro model using mouse microglial BV-2 cells. Upon 24 h of UVB exposure, BV-2 cells showed elevated oxidative stress markers and, cyclooxygenase (COX-2) expression, enhanced phagocytic and chemotactic activities, along with an altered immune profiling. Notably, UVB exposure led to a selective increase in expression and activity of fatty acid amide hydrolase (FAAH), the main enzyme responsible for degradation of fatty acid ethanolamides. Pharmacological FAAH inhibition via URB597 counteracted the effects of UVB exposure, decreasing tumor necrosis factor α (TNF-α) and nitric oxide (NO) release and reverting reactive oxidative species (ROS), interleukin-1ß (IL-1ß), and interleukin-10 (IL-10) levels to the control levels. Our findings support the potential of enhanced fatty acid amide signaling in mitigating UVB-induced cellular damage, paving the way to further exploration of these lipids in light-induced immune responses.

3.
Mikrochim Acta ; 190(8): 306, 2023 07 19.
Artículo en Inglés | MEDLINE | ID: mdl-37466678

RESUMEN

An electrochemical impedimetric biosensor for human serum albumin (HSA) determination is proposed. The biosensor is based on water-phase assembled nanocomposites made of 2D WS2 nanoflakes and Au nanoparticles (AuNPs). The WS2 has been produced using a liquid-phase exfoliation strategy assisted by sodium cholate, obtaining a water-stable suspension that allowed the straightforward decoration with AuNPs directly in the aqueous phase. The resulting WS2/Au nanocomposite has been characterized by atomic force microscopy and Raman spectroscopy and, then, employed to modify screen-printed electrodes. Good electron-transfer features have been achieved. An electrochemical immunosensing platform has been assembled exploiting cysteamine-glutaraldehyde covalent chemistry for antibody (Ab) immobilization. The resulting immunosensor exhibited good sensitivity for HSA detection (LOD = 2 ng mL-1), with extended linear range (0.005 - 100 µg mL-1), providing a useful analytical tool for HSA determination in urine at relevant clinical ranges for microalbuminuria screening. The HSA quantification in human urine samples resulted in recoveries from 91.8 to 112.4% and was also reproducible (RSD < 7.5%, n = 3), with marked selectivity. This nanocomposite, thanks to the reliable performance and the ease of the assembling strategy, is a promising alternative for electrochemical immunosensing of health relevant markers.


Asunto(s)
Técnicas Biosensibles , Nanopartículas del Metal , Nanocompuestos , Humanos , Nanopartículas del Metal/química , Agua , Oro/química , Técnicas Biosensibles/métodos , Inmunoensayo/métodos , Albúmina Sérica Humana , Nanocompuestos/química
4.
Food Chem ; 420: 136112, 2023 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-37059022

RESUMEN

An electroanalytical lab-on-a-strip device for the direct extra-virgin olive oil (EVOO) antioxidant capacity evaluation is proposed. The lab-made device is composed of a CO2 laser nanodecorated sensor combined with a cutter-plotter molded paper-strip designed for EVOOs sampling and extraction. Satisfactory performance towards the most representative o-diphenols of EVOOs i.e., hydroxytyrosol (HY) and oleuropein (OL) were achieved; good sensitivity (LODHY = 2 µM; LODOL = 0.6 µM), extended linear ranges (HY: 10-250 µM; OL: 2.5-50 µM) and outstanding reproducibility (RSD < 5%, n = 3) were obtained in rectified oil. The device was challenged for the extraction-free analysis of 15 different EVOO samples, with satisfactory recoveries (90-94%; RSD < 5%, n = 3) and correlation with classical photometric assays (r ≥ 0.91). The proposed device includes all analysis steps, needs 4 µL of sample, and returns reliable results in 2 min, resulting portable and usable with a smartphone.


Asunto(s)
Antioxidantes , Rayos Láser , Aceite de Oliva/análisis , Antioxidantes/análisis , Reproducibilidad de los Resultados
5.
Anal Methods ; 15(10): 1250-1253, 2023 03 09.
Artículo en Inglés | MEDLINE | ID: mdl-36861684

RESUMEN

The development of ultrasensitive analytical detection methods for organophosphorus pesticides such as dimethoate (DMT) plays a key role in healthy food production. DMT is an inhibitor of acetylcholinesterase (AChE), which can lead to the accumulation of acetylcholine and result in symptoms related to the autonomous and central nervous systems. Herein, we report the first spectroscopic and electrochemical study on template removal after an imprinting process from a polypyrrole-based molecularly imprinted polymer (PPy-MIP) film for the detection of DMT. Several template removal procedures were tested and evaluated using X-ray photoelectron spectroscopy. The most effective procedure was achieved in 100 mM NaOH. The proposed DMT PPy-MIP sensor exhibits a limit of detection of (8 ± 2) × 10-12 M.


Asunto(s)
Impresión Molecular , Plaguicidas , Polímeros/química , Dimetoato , Pirroles/química , Impresión Molecular/métodos , Acetilcolinesterasa , Compuestos Organofosforados
6.
Foods ; 12(6)2023 Mar 17.
Artículo en Inglés | MEDLINE | ID: mdl-36981218

RESUMEN

The transition to a sustainable economic and environmental management of olive oil sector needs to be implemented in both national and regional territories through the introduction and development of innovative growing systems and variety. In this study, the olive oil quality parameters of local and allochthonous varieties cultivated in different orchards located in the Abruzzo region (Italy), using traditional and super high-density systems, were analyzed. Frantene, Lecciana, Koroneiki, and a mix of Arbequina and Lecciana provided olive oils rich in flavonoids and secoiridoids compounds with respect to the local varieties Frantoio, Leccino, and a mix of Dritta, Leccino, and Pendolino. Oleic/linoleic ratio was influenced by cultivar and training systems with super high-density olive oils rich in oleic acid. Frantene showed a peculiar fatty acid profile different from cultivars grown in the same location; moreover, interesting similarities were found between Frantene and the mix of Dritta, Leccino, and Pendolino in terms of health-related compounds. The potential development of innovative sustainable training system to improve olive oil quality was highlighted. The study's results identify olive varieties suitable for super high-density systems spread in the Abruzzo region, representing a valid alternative for the olive growers to improve both the quality of the olive oil, as well as the company's income.

7.
Nanomaterials (Basel) ; 13(3)2023 Jan 17.
Artículo en Inglés | MEDLINE | ID: mdl-36770338

RESUMEN

The demand for next-generation multifunctional nanovectors, combining therapeutic effects with specific cellular targeting, has significantly grown during the last few years, pursuing less invasive therapy strategies. Polyphenol-conjugated silver nanoparticles (AgNPs) appear as potential multifunctional nanovectors, integrating the biorecognition capability and the antioxidant power of polyphenols, the antimicrobial activity of silver, and the drug delivery capability of NPs. We present a spectroscopic and microscopic investigation on polyphenol-synthesized AgNPs, selecting caffeic acid (CA) and catechol (CT) as model polyphenols and using them as reducing agents for the AgNP green synthesis, both in the presence and in the absence of a capping agent. We exploit the plasmonic properties of AgNPs to collect Surface-Enhanced Raman Scattering (SERS) spectra from the nanosized region next to the Ag surface and to characterize the molecular environment in the proximity of the NP, assessing the orientation and tunable deprotonation level of CA, depending on the synthesis conditions. Our results suggest that the SERS investigation of such nanovectors can provide crucial information for their perspective biomedical application.

8.
Nat Prod Res ; 37(15): 2591-2595, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-35289674

RESUMEN

In this work, the chemical composition and the antioxidant evaluation of the inflorescences from 12 Cannabis sativa L. monoecious cultivars (Carmagnola Lemon CL, Ferimon F, Gran Sasso Kush GSK, Antal A, Carmagnola C, Kompolti K, Futura 75 F75, Villanova V, Tiborzallasi T, Finola FL, Kc Virtus KV and Pineapple P) cultivated at the same condition, were investigated. GC-MS analysis was carried out to evaluate the volatile fraction, while HPLC-MS/MS was used for cannabinoids and polyphenolic compounds. The evaluation of antioxidant activity was carried out using ABTS*+, Trolox equivalence antioxidant capacity (TEAC), ferric reducing antioxidant property (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH*) assays in vitro. The obtained data, demonstrated that each cultivar has a characteristic chemical profile, with highest antioxidant capacity for CL, F75, GSK and F. Based on the in vitro antioxidant activity the plant extracts can be considered as promising candidates for different applications in food field.


Asunto(s)
Cannabinoides , Cannabis , Cannabis/química , Antioxidantes/análisis , Espectrometría de Masas en Tándem , Cannabinoides/química , Extractos Vegetales/análisis
9.
Biosens Bioelectron ; 218: 114749, 2022 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-36183581

RESUMEN

Rapid volatile organic compounds (VOCs) detection is a hot topic today; in this framework nanomaterials and their tailorable chemistry offer a plethora of compelling opportunities. In this work, Group VI transition metal dichalcogenides (TMDs, i.e., MoS2, WSe2, MoSe2, and WSe2) were functionalized with organic compounds (ellagic acid, tannic acid, catechin, and sodium cholate) able to assist their sonochemical exfoliation in water. The 16 resulting water-dispersed 2D hybrid inorganic/organic TMDs resulted in a few-layer nanoflakes conformation and were used to modify quartz crystal microbalances (QCMs) to equip an e-nose for VOCs determination. The ability of the sensors for the detection of VOCs was assessed on alcohols, terpenes, esters, and aldehydes; the responses were significatively different, confirming the synergic effect of TMD and the organic compound in the interaction with VOCs. The 16 sensors exhibited quantitative responses for VOCs (R2≥0.978) with fast signals recovery (<100 s) and repeatable (RSD ≤9.3%, n = 5), reproducible (RSD ≤12.8%, n = 3) and stable (RSD ≤14.6%, 3 months) signals. As proof of applicability, in an e-nose format, banana aroma evolution during post-harvest ripening was successfully monitored using the 2D TMDs-based sensors array. These data demonstrate that TMDs exfoliated in water with different organic compounds are sustainable functional nanomaterials, able to offer new opportunities in nano-bioelectronic applications.


Asunto(s)
Técnicas Biosensibles , Catequina , Elementos de Transición , Compuestos Orgánicos Volátiles , Nariz Electrónica , Molibdeno/química , Colato de Sodio , Elementos de Transición/química , Agua/química , Aldehídos , Taninos , Terpenos
10.
Antioxidants (Basel) ; 11(10)2022 Oct 11.
Artículo en Inglés | MEDLINE | ID: mdl-36290731

RESUMEN

A new sustainable route to nanodispersed and functionalized carbon black in water phase (W-CB) is proposed. The sonochemical strategy exploits ultrasounds to disaggregate the CB, while two selected functional naturally derived compounds, sodium cholate (SC) and rosmarinic acid (RA), act as stabilizing agents ensuring dispersibility in water adhering onto the CB nanoparticles' surface. Strategically, the CB-RA compound is used to drive the AuNPs self-assembling at room temperature, resulting in a CB surface that is nanodecorated; further, this is achieved without the need for additional reagents. Electrochemical sensors based on the proposed nanomaterials are realized and characterized both morphologically and electrochemically. The W-CBs' electroanalytical potential is proved in the anodic and cathodic window using caffeic acid (CF) and hydroquinone (HQ), two antioxidant compounds that are significant for food and the environment. For both antioxidants, repeatable (RSD ≤ 3.3%; n = 10) and reproducible (RSD ≤ 3.8%; n = 3) electroanalysis results were obtained, achieving nanomolar detection limits (CF: 29 nM; HQ: 44 nM). CF and HQ are successfully determined in food and environmental samples (recoveries 97-113%), and also in the presence of other phenolic classes and HQ structural isomers. The water dispersibility of the proposed materials can be an opportunity for (bio) sensor fabrication and sustainable device realization.

11.
Molecules ; 27(10)2022 May 11.
Artículo en Inglés | MEDLINE | ID: mdl-35630565

RESUMEN

Polyphenols (PCs) are a numerous class of bioactive molecules and are known for their antioxidant activity. In this work, the potential of the quadrupole/linear ion trap hybrid mass spectrometer (LIT-QqQ) was exploited to develop a semi-untargeted method for the identification of polyphenols in different food matrices: green coffee, Crocus sativus L. (saffron) and Humulus lupulus L. (hop). Several conjugate forms of flavonoids and hydroxycinnamic acid were detected using neutral loss (NL) as a survey scan coupled with dependent scans with enhanced product ion (EPI) based on information-dependent acquisition (IDA) criteria. The presented approach is focused on a specific class of molecules and provides comprehensive information on the different conjugation models that are related to specific base molecules, thus allowing a quick and effective identification of all possible combinations, such as mono-, di-, or tri-glycosylation or another type of conjugation such as quinic acid esters.


Asunto(s)
Polifenoles , Espectrometría de Masas en Tándem , Cromatografía Liquida/métodos , Fenoles
12.
Anal Chim Acta ; 1183: 338971, 2021 Oct 23.
Artículo en Inglés | MEDLINE | ID: mdl-34627526

RESUMEN

In this work, a low-cost, disposable, and portable lab-on-paper device is proposed to simultaneously quantify total polyphenol content (TPC) and antioxidant capacity (AOC) in 15 min; the assay requires no pre-treatment of the samples. The lab-on-paper device fabrication has been carried out employing a xurography-based benchtop microfabrication technology using low-cost materials as chromatography paper and polymeric sheets. Extra virgin olive oil (EVOO) phenolic compounds' represents a nutritional added value, nevertheless, the high lipidic content hinders their direct and rapid analysis, resulting in an extremely challenging sample. The realized lab-on-paper allows to perform the dual TPC and AOC determination in three simple steps: (i) sample loading, (ii) analytes transport to the analysis spot, and (iii) double colorimetric analysis exploiting the growth of AuNPs and AgNPs on paper mediated by phenolic compounds. Signal acquisition is achieved using a standard digital camera. The dual colorimetric assay is able to detect phenolic compounds in the 25-500 mg L-1 range with limits of detection ≤6 mg L-1 and good reproducibility (RSDs ≤11%). Direct analysis of EVOO samples (n = 30) correlated well (r > 0.92) with conventional spectrophotometric methods for TPC and AOC determination.


Asunto(s)
Oro , Nanopartículas del Metal , Aceite de Oliva , Aceites de Plantas , Reproducibilidad de los Resultados
13.
Mikrochim Acta ; 188(11): 369, 2021 10 07.
Artículo en Inglés | MEDLINE | ID: mdl-34618244

RESUMEN

A new green and effective sonochemical liquid-phase exfoliation (LPE) is proposed wherein a flavonoid compound, catechin (CT), promotes the formation of conductive, redox-active, water-phase stable graphene nanoflakes (GF). To maximize the GF-CT redox activity, the CT concentration and sonication time have been studied, and the best performing nanomaterial-fraction selected. Physicochemical and electrochemical methods have been employed to characterize the morphological, structural, and electrochemical features of the GF-CT nanoflakes. The obtained GF intercalated with CT exhibits fully reversible electrochemistry (ΔEp = 28 mV, ipa/ipc = ⁓1) because of the catecholic adducts. GF-CT-integrated electrochemistry was generated directly during LPE of graphite, with no need of graphene oxide production, nor activation steps, electropolymerization, or ex-post functionalization. The GF-CT electro-mediator ability has been proven towards hydrazine (HY) and ß-nicotinamide adenine dinucleotide (NADH) by simply drop-casting the redox-material onto screen-printed electrodes. GF-CT-based electrodes by using amperometry exhibited high sensitivity and extended linear ranges (HY: LOD = 0.1 µM, L.R. 0.5-150 µM; NADH: LOD = 0.6 µM, L.R. 2.5-200 µM) at low overpotential (+ 0.15 V) with no electrode fouling. The GF-CT electrodes are performing significantly better than commercial graphite electrodes and graphene nanoflakes exfoliated with a conventional surfactant, such as sodium cholate. Recoveries of 94-107% with RSD ≤ 8% (n = 3) for determination of HY and NADH in environmental and biological samples were achieved, proving the material functionality also in challenging analytical media. The presented GF-CT is a new functional redox-active material obtainable with a single-pot sustainable strategy, exhibiting standout properties particularly prone to (bio)sensors and cutting-edge device development.


Asunto(s)
Grafito
14.
Biosens Bioelectron ; 191: 113471, 2021 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-34246123

RESUMEN

The design of a new class of selective and high affinity antibody mimetics termed clamp peptide (CP) that incorporate three short peptides structurally and mechanically mimicking a clamp is proposed as sensing elements for a reliable detection sensor platform. The CPs consist of two short peptides functioning as arms that recognize two different epitopes in the target protein and are connected by a third short peptide that acts as a hinge between the peptide arms. For the construction of CPs, we employed a rational design combined with computational methods. To illustrate our approach, we designed a CP that binds selectively to the envelope protein of the Zika virus (ZIKV). The virtual docking cycles were run maximizing the discrimination between ZIKV and Dengue virus (DENV) envelope proteins. DENV was chosen among the flavivirus family because it has high structural similarity with ZIKV. When employed in a colorimetric binding assay or in label-free electrochemical impedance sensor format, the CP was selective for ZIKV vs DENV particles showing detection limit under 104 copies/mL, comparable to anti-ZIKV antibodies. Apparent dissociation binding constants (Kd) confirmed a better performance of CPs than mono-arm peptides (Kd of best CP = 162 nM ± 23 nM; Kd of best mono-arm peptide = 11.15 ± 2.76 µM). The performance of the assays based on CPs was also verified in serum and urine (diluted 1:10 and 1:1 respectively). The detection limits of CPs decreased about one order of magnitude for ZIKV detection in serum or urine, with a distinct analytical signal starting from 105 copies/mL of ZIKV.


Asunto(s)
Técnicas Biosensibles , Virus del Dengue , Dengue , Infección por el Virus Zika , Virus Zika , Anticuerpos Antivirales , Reacciones Cruzadas , Humanos , Péptidos , Infección por el Virus Zika/diagnóstico
15.
J Chromatogr A ; 1651: 462315, 2021 Aug 16.
Artículo en Inglés | MEDLINE | ID: mdl-34157475

RESUMEN

In this work two different acquisition approaches were used for the quantification and/or tentative identification of phenolic compounds (PCs) in plant matrices by HPLC-MS/MS. A targeted approach, based on MRM acquisition mode, was used for the identification and quantification of a list of target analytes by comparison with standards; a semi-targeted approach was also developed by the precursor ion scan and neutral loss for the tentative identification of compounds not included in the target list. Analysis of phenolic content in three different plant matrices (curry leaves, hemp and blueberry) was carried out. The extraction and clean-up steps were set up according to the characteristics of the sample allowing to minimize the interfering compounds present in such complex matrices, as proved by the low matrix effect obtained (<16%) and recovery values ranging from 45% to 98% for all the analytes. This approach provided a sensitive and robust quantitative analysis of the target compounds with LOQs between 0.0002 and 0.05 ng mg-1, which allowed the identification and quantification of several hydroxycinnamic and hydroxybenzoic acids, in addition to numerous flavonoids in all three matrices. Furthermore, different moieties were considered as neutral losses or as precursor ions in semi-targeted MS/MS approach, providing the putative identification of different glycosylated forms of flavonoids, such as luteolin-galactoside and diosmin in all three matrices, while apigenin-glucuronide was detected in hemp and quercetin-glucuronide in blueberry. A further study was carried out by MS3, allowing the discrimination of compounds with similar aglycones, such as luteolin and kaempferol.


Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Fenoles/análisis , Plantas/química , Espectrometría de Masas en Tándem/métodos , Hojas de la Planta/química , Reproducibilidad de los Resultados , Espectrometría de Masa por Ionización de Electrospray/métodos
16.
Colloids Surf B Biointerfaces ; 199: 111533, 2021 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-33388719

RESUMEN

An exponential increase of scientific works dealing with the use of polyphenol-rich 'natural products' for the synthesis of bioactive AgNPs is in progress. However, a lack of fundamental studies on phytochemical compounds involved, and their role is evident. In this work, a comprehensive study of the antifungal performances of silver nanoparticles (AgNPs) synthesized exclusively with phenolic compounds (PCs) with different structures and different antioxidant capacity is presented. The experimental hypothesis is that AgNPs@PCs produced with different PCs can exert different toxicity. In particular, di-hydroxylic and tri-hydroxylic phenolic acids (caffeic acid and gallic acid) and flavonoids (catechin and myricetin) were compared. A room temperature rapid and simple AgNPs synthesis was carefully optimized, obtaining stable and reproducible colloids. AgNPs@PCs suspensions were characterized by UV-vis spectroscopy, ς-potential, dynamic light scattering and transmission electron microscopy. AgNPs@PCs radical scavenging capacity was also assessed. Finally, the AgNPs@PCs antifungal effect was tested against Aspergillus niger, particularly on spore germination and mycelial growth. The different antifungal activity was attributed to the different PCs' ability to generate/stabilize AgNPs with different shells, residual antioxidant capacity, and capacity to interact and aggregate during their 'attack' to A. niger hyphae. This work paves the way for the rational use of PCs and PCs rich-products for AgNPs-based applications.


Asunto(s)
Nanopartículas del Metal , Plata , Antifúngicos/farmacología , Aspergillus niger , Extractos Vegetales , Plata/farmacología
17.
Biosens Bioelectron ; 170: 112669, 2020 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-33035897

RESUMEN

A Prussian blue-based electrode array (PBEA) constituted by eight stencil-printed electrodes on a flexible PET (polyethylene terephthalate) substrate is proposed for in-situ HeLa cell culturing and real-time detection of the released H2O2. The array was suitably interfaced with a poly- (methyl methacrylate) (PMMA) well-containing holders resulting in a low cost multichambered chip. PBEA fabrication was carried out employing a xurography-based cost-effective benchtop microfabrication technology using just a desktop cutting plotter and office grade thermal-laminator. The hydrophobicity of the PET isolating layer allows to constrain cell-containing drops directly on top of the electrochemical cells. HeLa cells growth in the very close vicinity of the working electrode ensures in-situ cell seeding, incubation, and further electrochemical detection of the H2O2 released, enabling high-throughput analysis. Selective and sensitive electrochemical sensing of hydrogen peroxide was carried out at -100 mV vs Ag|AgCl; the resulting LOD was 1.9 µM. Remarkably, the analytical exploitability of the approach was demonstrated by detection of the hydrogen peroxide released from HeLa cells stimulated with N-Formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP) and after pretreatment of the cells with cocoa polyphenols, that induced a decreased oxidative stress levels. These data make our approach a promising tool for oxidative stress evaluation in cell cultures and biological systems.


Asunto(s)
Técnicas Biosensibles , Peróxido de Hidrógeno , Electrodos , Ferrocianuros , Células HeLa , Humanos , Estrés Oxidativo
18.
Anal Chem ; 92(19): 13565-13572, 2020 10 06.
Artículo en Inglés | MEDLINE | ID: mdl-32869640

RESUMEN

A novel benchtop approach to fabricate xurography-enabled thermally transferred (XTT) carbon nanomaterial-based electrochemical sensors is proposed. Filtered nanomaterial (NM) films were transferred from Teflon filters to polyethylene terephthalate-ethylene vinyl acetate (PET-EVA) substrates by a temperature-driven approach. Customized PET-EVA components were xurographically patterned by a cutting plotter. The smart design of PET-EVA films enabled us to selectively transfer the nanomaterial to the exposed EVA side of the substrate. Hence, the substrate played an active role in selectively controlling where nanomaterial transfer occurred allowing us to design different working electrode geometries. Counter and reference electrodes were integrated by a stencil-printing approach, and the whole device was assembled by thermal lamination. To prove the versatility of the technology, XTT materials were exclusively made of carbon black (XTT-CB), multiwalled carbon nanotubes (XTT-MWCNTs), and single-walled carbon nanotubes (XTT-SWCNTs). Their electrochemical behavior was carefully studied and was found to be highly dependent on the amount and type of NM employed. XTT-SWCNTs were demonstrated to be the best-performing sensors, and they were employed for the determination of l-tyrosine (l-Tyr) in human plasma from tyrosinemia-diagnosed patients. High analytical performance toward l-Tyr (linear range of 0.5-100 µM, LOD = 0.1 µM), interelectrode precision (RSD ip,a = 3%, n = 10; RSD calibration slope = 4%, n = 3), and accurate l-Tyr quantification in plasma samples with low relative errors (≤7%) compared to the clinical declared values were obtained. The proposed benchtop approach is cost-effective and straightforward, does not require sophisticated facilities, and can be potentially employed to develop pure or hybrid nanomaterial-based electrodes.


Asunto(s)
Carbono/química , Técnicas Electroquímicas , Nanoestructuras/química , Tereftalatos Polietilenos/análisis , Polivinilos/análisis , Temperatura
19.
Foods ; 9(9)2020 Sep 02.
Artículo en Inglés | MEDLINE | ID: mdl-32887367

RESUMEN

Appropriate and standardized techniques for the extraction of secondary metabolites with interesting biological activity from plants are required. In this work, a comparison of different conventional and unconventional extraction techniques (maceration-M, Soxhlet-S, ultrasound assisted extraction-UAE, and rapid solid-liquid dynamic extraction-RSLDE) was investigated. Bioactive compounds were extracted from Thymus vulgaris L. (thyme), Cannabis sativa L. (industrial hemp) and Coriandrum sativum L. (coriander) and chemically characterized for their volatile fraction and polyphenolic content by means of gas chromatography-mass spectrometry (GC-MS) and high performance liquid chromatography-ultraviolet (HPLC-UV). Linalool (48.19%, RSLDE) and carvacrol (21.30%, M) for thyme, caryophyllene (54.78%, S) and humulene (14.13%, S) for hemp, and linalool (84.16%, RSLDE) for coriander seeds were the main compounds among terpenes, while thyme was the richest source of polyphenols with rosmarinic acid (51.7 mg/g dry extract-S), apigenin (7.6 mg/g dry extract-S), and luteolin (4.1 mg/g dry extract-UAE) being the most abundant. In order to shed light on their potential as natural food preservatives, the biological activity of the extracts was assessed in terms of antioxidant activity (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid-ABTS˙+, ferric reducing antioxidant power-FRAP, 2,2-diphenyl-1-picrylhydrazyl-DPPH˙ assays) and phenolic content (Folin-Ciocâlteu method). For thyme, Soxhlet extracts showed best performances in FRAP and ABTS˙+ assays (74 mg TE/g dry extract and 134 mg TE/g dry extract, respectively), while Soxhlet and RSLDE extracts recorded similar activity in DPPH˙ (107-109 mg TE/g dry extract). For hemp and coriander, indeed, RSLDE extracts accounted for higher antioxidant activity as evidenced by FRAP (80 mg TE/g dry extract and 18 mg TE/g dry extract, respectively) and ABTS˙+ (557 mg TE/g dry extract and 48 mg TE/g dry extract, respectively) assays. With respect to DPPH˙, the best results were observed for UAE extracts (45 mg TE/g dry extract and 220 mg TE/g dry extract, respectively). Our findings suggest that all the investigated techniques are valid extraction methods to retain bioactive compounds and preserve their activity for application in food and pharmaceutical formulations. Among them, the innovative RSLDE stands out for the slightly higher antioxidant performances of the extracts, coupled with the facility of use and standardization of the extraction process.

20.
Sensors (Basel) ; 20(16)2020 Aug 08.
Artículo en Inglés | MEDLINE | ID: mdl-32784423

RESUMEN

Detection and monitoring of volatiles is a challenging and fascinating issue in environmental analysis, agriculture and food quality, process control in industry, as well as in 'point of care' diagnostics. Gas chromatographic approaches remain the reference method for the analysis of volatile organic compounds (VOCs); however, gas sensors (GSs), with their advantages of low cost and no or very little sample preparation, have become a reality. Gas sensors can be used singularly or in array format (e.g., e-noses); coupling data output with multivariate statical treatment allows un-target analysis of samples headspace. Within this frame, the use of new binding elements as recognition/interaction elements in gas sensing is a challenging hot-topic that allowed unexpected advancement. In this review, the latest development of gas sensors and gas sensor arrays, realized using peptides, molecularly imprinted polymers and DNA is reported. This work is focused on the description of the strategies used for the GSs development, the sensing elements function, the sensors array set-up, and the application in real cases.


Asunto(s)
ADN/química , Nariz Electrónica , Gases/análisis , Impresión Molecular , Péptidos/química , Compuestos Orgánicos Volátiles/análisis , Polímeros
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