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1.
ACS Sens ; 8(1): 353-362, 2023 01 27.
Artigo em Inglês | MEDLINE | ID: mdl-36599088

RESUMO

Pseudomonas aeruginosa is a ubiquitous multi-drug-resistant bacterium, capable of causing serious illnesses and infections. This research focuses on the development of a thermal sensor for the indirect detection of P. aeruginosa infection using molecularly imprinted polymers (MIPs). This was achieved by developing MIPs for the detection of pyocyanin, the main toxin secreted by P. aeruginosa. To this end, phenazine was used as a dummy template, evaluating several polymeric compositions to achieve a selective MIP for pyocyanin recognition. The sensitivity of the synthesized MIPs was investigated by UV-vis analysis, with the best composition having a maximum rebinding capacity of 30 µmol g-1 and an imprinting factor (IF) of 1.59. Subsequently, the MIP particles were immobilized onto planar aluminum chips using an adhesive layer, to perform thermal resistance measurements at clinically relevant concentrations of pyocyanin (1.4-9.8 µM), achieving a limit of detection (LoD) of 0.347 ± 0.027 µM. The selectivity of the sensor was also scrutinized by subjecting the receptor to potential interferents. Furthermore, the rebinding was demonstrated in King's A medium, highlighting the potential of the sensor for the indirect detection of P. aeruginosa in complex fluids. The research culminates in the demonstration of the MIP-based sensor's applicability for clinical diagnosis. To achieve this goal, an experiment was performed in which the sensor was exposed to pyocyanin-spiked saliva samples, achieving a limit of detection of 0.569 ± 0.063 µM and demonstrating that this technology is suitable to detect the presence of the toxin even at the very first stage of its production.


Assuntos
Impressão Molecular , Polímeros Molecularmente Impressos , Pseudomonas aeruginosa , Piocianina , Técnicas Eletroquímicas
2.
Food Chem ; 404(Pt B): 134653, 2023 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-36327513

RESUMO

Accurate and fast on-site detection of harmful microorganisms in food products is a key preventive step to avoid food-borne illness and product recall. In this study, screen-printed electrodes (SPEs) were functionalized via a facile strategy with surface imprinted polymers (SIPs). The SIP-coated SPEs were used in combination with the heat transfer method (HTM) for the real-time detection of Escherichia coli. The sensor was tested in buffer, with a reproducible and sensitive response that attained a limit of detection of 180 CFU/mL. Furthermore, selectivity was assessed by analyzing the sensor's response to C. sakazakii, K. pneumoniae and S. aureus as analogue strains. Finally, the device was successfully used for the detection of E. coli in spiked milk as proof-of-application, requiring no additional sample preparation. These results suggest the proposed thermal biosensor possesses the potential of becoming a tool for routine, on-site monitoring of E. coli in food safety applications.


Assuntos
Técnicas Biossensoriais , Escherichia coli , Staphylococcus aureus , Eletrodos , Técnicas Biossensoriais/métodos , Laticínios , Limite de Detecção
3.
Foods ; 11(18)2022 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-36141032

RESUMO

In recent years, melamine-sensing technologies have increasingly gained attention, mainly due to the misuse of the molecule as an adulterant in milk and other foods. Molecularly imprinted polymers (MIPs) are ideal candidates for the recognition of melamine in real-life samples. The prepared MIP particles were incorporated into a thermally conductive layer via micro-contact deposition and its response towards melamine was analyzed using the heat-transfer method (HTM). The sensor displayed an excellent selectivity when analyzing the thermal response to other chemicals commonly found in foods, and its applicability in food safety was demonstrated after evaluation in untreated milk samples, demonstrating a limit of detection of 6.02 µM. As the EU/US melamine legal limit in milk of 2.5 mg/kg falls within the linear range of the sensor, it can offer an innovative solution for routine screening of milk samples in order to detect adulteration with melamine. The results shown in this work thus demonstrate the great potential of a low-cost thermal platform for the detection of food adulteration in complex matrices.

4.
ACS Sens ; 6(12): 4515-4525, 2021 12 24.
Artigo em Inglês | MEDLINE | ID: mdl-34825565

RESUMO

Glucose bio-sensing technologies have received increasing attention in the last few decades, primarily due to the fundamental role that glucose metabolism plays in diseases (e.g., diabetes). Molecularly imprinted polymers (MIPs) could offer an alternative means of analysis to a field that is traditionally dominated by enzyme-based devices, posing superior chemical stability, cost-effectiveness, and ease of fabrication. Their integration into sensing devices as recognition elements has been extensively studied with different readout methods such as quartz-crystal microbalance or impedance spectroscopy. In this work, a dummy imprinting approach is introduced, describing the synthesis and optimization of a MIP toward the sensing of glucose. Integration of this polymer into a thermally conductive receptor layer was achieved by micro-contact deposition. In essence, the MIP particles are pressed into a polyvinyl chloride adhesive layer using a polydimethylsiloxane stamp. The prepared layer is then evaluated with the so-called heat-transfer method, allowing the determination of the specificity and the sensitivity of the receptor layer. Furthermore, the selectivity was assessed by analyzing the thermal response after infusion with increasing concentrations of different saccharide analogues in phosphate-buffered saline (PBS). The obtained results show a linear range of the sensor of 0.0194-0.3300 mM for the detection of glucose in PBS. Finally, a potential application of the sensor was demonstrated by exposing the receptor layer to increasing concentrations of glucose in human urine samples, demonstrating a linear range of 0.0444-0.3300 mM. The results obtained in this paper highlight the applicability of the sensor both in terms of non-invasive glucose monitoring and for the analysis of food samples.


Assuntos
Impressão Molecular , Glicemia , Automonitorização da Glicemia , Glucose , Humanos , Polímeros Molecularmente Impressos
5.
J Chem Educ ; 98(2): 439-444, 2021 Feb 09.
Artigo em Inglês | MEDLINE | ID: mdl-33583951

RESUMO

The need to develop interest in STEM (science, technology, engineering, and mathematics) skills in young pupils has driven many educational systems to include STEM as a subject in primary schools. In this work, a science kit aimed at children from 8 to 14 years old is presented as a support platform for an innovative and stimulating approach to STEM learning. The peculiar design of the kit, based on modular components, is aimed to help develop a multitude of skills in the young students, dividing the learning process into two phases. During phase 1 the pupils build the experimental setup and visualize the scientific phenomena, while in phase 2, they are introduced and challenged to understand the principles on which these phenomena are based, guided by a handbook. This approach aims at making the experience more inclusive, stimulating the interest and passion of the pupils for scientific subjects.

6.
Molecules ; 25(22)2020 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-33182534

RESUMO

The rapid sensing of drug compounds has traditionally relied on antibodies, enzymes and electrochemical reactions. These technologies can frequently produce false positives/negatives and require specific conditions to operate. Akin to antibodies, molecularly imprinted polymers (MIPs) are a more robust synthetic alternative with the ability to bind a target molecule with an affinity comparable to that of its natural counterparts. With this in mind, the research presented in this article introduces a facile MIP-based dye displacement assay for the detection of (±) amphetamine in urine. The selective nature of MIPs coupled with a displaceable dye enables the resulting low-cost assay to rapidly produce a clear visual confirmation of a target's presence, offering huge commercial potential. The following manuscript characterizes the proposed assay, drawing attention to various facets of the sensor design and optimization. To this end, synthesis of a MIP tailored towards amphetamine is described, scrutinizing the composition and selectivity (ibuprofen, naproxen, 2-methoxphenidine, quetiapine) of the reported synthetic receptor. Dye selection for the development of the displacement assay follows, proceeded by optimization of the displacement process by investigating the time taken and the amount of MIP powder required for optimum displacement. An optimized dose-response curve is then presented, introducing (±) amphetamine hydrochloride (0.01-1 mg mL-1) to the engineered sensor and determining the limit of detection (LoD). The research culminates in the assay being used for the analysis of spiked urine samples (amphetamine, ibuprofen, naproxen, 2-methoxphenidine, quetiapine, bupropion, pheniramine, bromopheniramine) and evaluating its potential as a low-cost, rapid and selective method of analysis.


Assuntos
Anfetaminas/urina , Corantes/química , Polímeros Molecularmente Impressos , Polímeros/química , Detecção do Abuso de Substâncias/métodos , Urina/química , Anfetamina/urina , Bromofeniramina/urina , Bupropiona/urina , Relação Dose-Resposta a Droga , Técnicas Eletroquímicas , Reações Falso-Positivas , Humanos , Ibuprofeno/urina , Limite de Detecção , Impressão Molecular , Naproxeno/urina , Feniramina/urina , Piperidinas/urina , Pós , Fumarato de Quetiapina/urina
7.
Bioorg Med Chem ; 28(13): 115529, 2020 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-32386952

RESUMO

The bacterial deacetylase LpxC is a promising target for the development of antibiotics selectively combating Gram-negative bacteria. To improve the biological activity of the reported benzyloxyacetohydroxamic acid 9 ((S)-N-hydroxy-2-{2-hydroxy-1-[4-(phenylethynyl)phenyl]ethoxy}acetamide), its hydroxy group was replaced by a triazole ring. Therefore, in divergent syntheses, triazole derivatives exhibiting rigid and flexible lipophilic side chains, different configurations at their stereocenter, and various substitution patterns at the triazole ring were synthesized, tested for antibacterial and LpxC inhibitory activity, and structure-activity relationships were deduced based on docking and binding energy calculations.


Assuntos
Amidoidrolases/antagonistas & inibidores , Antibacterianos/síntese química , Proteínas de Bactérias/antagonistas & inibidores , Inibidores Enzimáticos/síntese química , Ácidos Hidroxâmicos/síntese química , Triazóis/química , Antibacterianos/farmacologia , Reação de Cicloadição , Inibidores Enzimáticos/farmacologia , Bactérias Gram-Negativas/efeitos dos fármacos , Humanos , Ácidos Hidroxâmicos/farmacologia , Testes de Sensibilidade Microbiana , Simulação de Acoplamento Molecular , Estrutura Molecular , Estereoisomerismo , Relação Estrutura-Atividade
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