Percorrer por autor "Nouws, Henri"
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- Amperometric enzyme sensor for the rapid determination of histaminePublication . Torre, Ricarda; Costa Rama, Estefanía; LOPES, Paula; Nouws, Henri; Delerue-Matos, CristinaThe concentration of histamine, a biogenic amine, in fish is considered a fish spoilage indicator. Therefore, the development of a rapid-response and portable tool that allows its on-site analysis is very interesting since the consumption of high amounts of histamine causes scombroid poisoning. Thus, in this work a simple enzymatic sensor for the determination of histamine, based on a screen-printed carbon electrode and the enzyme diamine oxidase, was developed. The enzyme was immobilized on the electrode surface through a simple cross-linking procedure employing glutaraldehyde and bovine serum albumin. Chronoamperometry was used as the detection technique; the sensor showed a short response time (60 s, -0.3 V) and the measurements were performed by only using 40 μL of sample solution. The sensor provides an useful linear range (between 1 and 75 mg L-1 in fish extract), excellent reproducibility (RSD = 2.6%) and is reusable for up to 7 measurements. The feasibility of the sensor was tested performing histamine analysis in fish extracts achieving recovery values of 103%.
- Breast cancer biomarker (HER2-ECD) detection using a molecularly imprinted electrochemical sensorPublication . Pacheco, João; Rebelo, Patrícia; Freitas, Maria; Nouws, Henri; Delerue-Matos, CristinaThe extracellular domain of the human epidermal growth factor receptor 2 (HER2-ECD) is a protein breast cancer biomarker. Its quantification in peripheral blood could provide an important contribution to diagnostics and patient follow-up. In this work an electrochemical molecularly imprinted polymer (MIP) sensor for the quantification of HER2-ECD was developed. The MIP was electropolymerized by cyclic voltammetry using a solution containing phenol and HER2-ECD on a screen-printed gold electrode (AuSPE). The sensor was characterized by cyclic voltammetry and electrochemical impedance spectroscopy. The analysis of HER2-ECD was performed by differential pulse voltammetry using ([Fe(CN)6]3−/4−as redox probe. The linear range was established in the concentration interval from 10 to 70 ng/mL HER2-ECD, with a limit of detection of 1.6 ng/L and a limit of quantification of 5.2 ng/mL. Through the analysis of other protein biomarkers, the MIP sensor was found to be selective. Furthermore, these proteins did not interfere in the analysis of the selected biomarker. The developed sensor was used for the analysis of spiked human serum samples, providing adequate recovery values and precise results. The outcomes of this study indicate that the developed MIP sensor could be useful in the non-invasive analysis of HER2-ECD in breast cancer patients.
- Diamine oxidase-modified screen-printed electrode for the redox-mediated determination of histaminePublication . Torre, Ricarda; Costa Rama, Estefanía; Nouws, Henri; Delerue-Matos, CristinaHistamine is an important biogenic amine because of its role in immune responses and the regulation of physiological functions. It is also used as a food freshness indicator, so its maximum concentration in fish is legally regulated. Although several robust and sensitive methods for histamine detection are already available, it continues to be a challenge to develop simple and portable devices that allow rapid histamine screening at any point of the fish production chain. Thus, in this work, a simple, miniaturized and low-cost sensor for histamine analysis was developed. The construction of the sensor only takes 30 min and consists of the immobilization of the enzyme diamine oxidase on the surface of a screen-printed carbon electrode by cross-linking. The quantification of histamine was achieved by chronoamperometry (+ 0.2V,120 s) using hexacyanoferrate (III) as a redox mediator. This selective sensor provided a low limit of detection (0.97 mg L−1) and accurate and precise results and was successfully applied to the analysis of spiked tuna and mackerel extracts,obtaining recovery values of 99–100%. Moreover, the sensor shows good stability, maintaining 87.7% of its initial signalafter 35 days.
- Disposable electrochemical immunosensor for analysis of cystatin C, a CKD biomarkerPublication . LOPES, Paula; Costa Rama, Estefanía; Beirão, Idalina; Nouws, Henri; Santos-Silva, Alice; Delerue-Matos, CristinaSpecific monitoring of cystatin C (CysC) levels in biological fluids is critical for diagnosis, treatment and mechanistic understanding of a spectrum of diseases, particularly chronic kidney disease (CKD). Despite evidences that CysC correlates with the high risk and/or progression of CKD, its use in clinical practice is still scarce. In this context, we report the development of a simple and sensitive immunosensor for the detection of CysC. The biosensor combines the technology of cost-effective screen-printed electrodes with the high specificity of a sandwich immunoassay. Optimized conditions showed that the sensor operates in a linear range between 10 and 100 ng mL-1, with a detection limit and a sensitivity of 6.0 ng mL-1 and 6.4 ± 0.3 μA ng mL-1 cm-2, respectively. Moreover, the sensor provided precise results (RSD ≤ 6.2%) and the quantification of CysC in CKD serum samples revealed to be in agreement with the values obtained by a particle-enhanced nephelometric immunoassay. In this light, the proposed immunosensor qualifies for clinical application, constituting a step forward in the development of fast, sensitive and cost-effective diagnostic tools that can improve the current medical care settings of CKD patients.
- Electrochemical Biosensing in Cancer Diagnostics and Follow-upPublication . Freitas, Maria; Nouws, Henri; Delerue-Matos, CristinaIn cancer, screening and early detection are critical for the success of the patient's treatment and to increase the survival rate. The development of analytical tools for non‐invasive detection, through the analysis of cancer biomarkers, is imperative for disease diagnosis, treatment and follow‐up. Tumour biomarkers refer to substances or processes that, in clinical settings, are indicative of the presence of cancer in the body. These biomarkers can be detected using biosensors, that, because of their fast, accurate and point of care applicability, are prominent alternatives to the traditional methods. Moreover, the constant innovations in the biosensing field improve the determination of normal and/or elevated levels of tumour biomarkers in patients’ biological fluids (such as serum, plasma, whole blood, urine, etc.). Although several biomarkers (DNA, RNA, proteins, cells) are known, the detection of proteins and circulating tumour cells (CTCs) are the most commonly reported due to their approval as tumour biomarkers by the specialized entities and commonly accepted for diagnosis by medical and clinical teams. Therefore, electrochemical immunosensors and cytosensors are vastly described in this review, because of their fast, simple and accurate detection, the low sample volumes required, and the excellent limits of detection obtained. The biosensing strategies reported for the six most commonly diagnosed cancers (lung, breast, colorectal, prostate, liver and stomach) are summarized and the distinct phases of the sensors’ constructions (surface modification, antibody immobilization, immunochemical interactions, detection approach) and applications are discussed.
- Enrichment of Dehalococcoides mccartyi spp. from a municipal activated sludge during AQDS-mediated bioelectrochemical dechlorination of 1,2-dichloroethane to ethenePublication . Leitão, Patrícia; Rossetti, Simona; Danko, Anthony S.; Nouws, Henri; Aulenta, FedericoThe application of bioelectrochemical systems (BES) for the treatment of chloroethanes has been so far limited, in spite of the high frequency that these contaminants are detected at contaminated sites. This work studied the biodegradation of 1,2-dichloroethane (1,2-DCA) in a lab-scale BES, inoculated with a municipal activated sludge and operated under a range of conditions, spanning from oxidative to reductive, both in the presence and in the absence of the humic acid analogue anthraquinone-2,6-disulfonate (AQDS) as a redox mediator. The results showed stable dechlorination of 1,2-DCA to ethene (up to 65 ± 5 lmol/L d), when the BES was operated at a set potential of 300 mV vs. SHE, in the presence of AQDS. Sustained filled-and-draw operation resulted in the enrichment of Dehalococcoides mccartyi. The results of this work provide new insights into the applicability of BES for groundwater remediation and the potential interaction between biogeochemistry and 1,2-DCA in humics-rich contaminated aquifers.
- Food allergen control: Tropomyosin analysis through electrochemical immunosensingPublication . Torre, Ricarda; Freitas, Maria; Costa‐Rama, Estefanía; Nouws, Henri; Delerue-Matos, CristinaRegulations of the EU obliges the indication of the presence of allergens on food labels. This work reports the development of an electrochemical immunosensor to determine tropomyosin (TPM) – a major shellfish allergen – prevailing in the muscles of crustacean species. Two linear ranges between the signal and TPM concentration were obtained: between 2.5 and 20 ng mL−1 and between 30 and 200 ng mL−1, with a lowest limit of detection of 0.47 ng mL−1. The selectivity of the optimized immunoassay, tested with other food allergens (e.g., Cyp c 1, a fish allergen), assures the effective detection of TPM, enabling successful control of foodstuff labelling. Several (12) foods, containing high and low TPM concentrations and TPM-free samples, were analysed using the sensor. A conventional ELISA kit and recovery assays were used to evaluate the accuracy of the results.
- Impact of magnetite nanoparticles on the syntrophic dechlorination of 1,2-dichloroethanePublication . Leitão, Patrícia; Aulenta, Federico; Rossetti, Simona; Nouws, Henri; Danko, Anthony S.In anaerobic environments microorganisms exchange electrons with community members and with soil and groundwater compounds. Interspecies electron transfer (IET) occurs by several mechanisms: diffusion of redox compounds or direct contact between cells. This latter mechanism may be facilitated by the presence of conductive nanoparticles (NP), possibly serving as electron conduits among microorganisms. Our study examined the effect of magnetite (Fe3O4) NP on the dechlorination of 1,2-dichloroethane (1,2-DCA) by a mixed-culture. The addition of NP (170 mg L− 1 total Fe) enhanced the acetate-driven reductive dechlorination of 1,2-DCA to harmless ethene (via reductive dihaloelimination) up to 3.3-times (2.3 μeq L− 1 d− 1 vs. 0.7 μeq L− 1 d− 1), while decreasing the lag time by 0.8 times (23 days) when compared to unamended (magnetite-free) microcosms. Dechlorination activity was correlated with the abundance of Dehalococcoides mccartyi, which accounted up to 50% of total bacteria as quantified by CARD-FISH analysis, pointing to a key role of this microorganism in the process. Given the widespread abundance of conductive minerals in the environment, the results of this study may provide new insights into the fate of 1,2-DCA and suggest new tools for its remediation by linking biogeochemical mechanisms.
- Molecularly imprinted electrochemical sensor for the point-of-care detection of a breast cancer biomarker (CA 15-3)Publication . Pacheco, João; Silva, Marta S.V.; Freitas, Maria; Nouws, Henri; Delerue-Matos, CristinaThe incidence of breast cancer has been increasing over the years. To control and monitor this disease several tumor biomarkers have been proposed for early diagnosis, patient follow-up and/or treatment guidance. The only serum breast cancer biomarker in current use is the cancer antigen 15-3 (CA 15-3). In this work a molecularly imprinted polymer (MIP)-based electrochemical (voltammetric) sensor to monitor breast cancer was developed, based on direct surface imprinting of CA 15-3 on a screen-printed gold electrode (Au-SPE). The imprinting was performed in two steps: (1) adsorption of CA 15-3 on the surface of the Au-SPE and (2) electropolymerization of 2-aminophenol around the adsorbed protein. After extraction of the imprinted protein voltammetric analysis was conducted using hexacyanoferrate(II/III) as redox probe, measuring the signals before and after protein binding. The sensor was characterized by voltammetric techniques and electrochemical impedance spectroscopy, and the analytical responses of imprinted and non-imprinted polymer sensors were studied. A linear relationship between the peak current intensity of the redox probe and the logarithm of CA 15-3 concentration was established between 5 and 50 U mL−1, achieving a limit of detection of 1.5 U mL−1. The prepared MIP-sensor provides fast (15 min) analysis and is cheap, easy to prepare, disposable and could easily be integrated in small portable point-of care devices.
- Natural Magnetite Minerals Enhance 1,2-Dichloroethane Reductive DechlorinationPublication . Leitão, Patrícia; Tucci, Matteo; Cruz Viggi, Carolina; Nouws, Henri; Danko, Anthony S.; Aulenta, FedericoContamination of soil and groundwater by chlorinated solvents is an environmental issue of primary concern. Recently, electrically conductive iron particles have been proposed as a novel approach to accelerate anaerobic bioremediation processes. In fact, it was demonstrated that conductive particles facilitate the exchange of electrons between microorganisms via Direct Interspecies Electron Transfer (DIET) processes, thus enhancing the pollutant-degrading potential of the microbial community. However, the use of natural minerals in this context has not been reported so far. In this study, we applied, for the first time, natural magnetite and hematite to accelerate the reductive dechlorination of 1,2-dichloroethane by an enrichment culture in lab-scale anaerobic microcosms. After four feeding cycles, low magnetite-amended microcosms (13 mg/L) yielded the highest rate of 1,2-DCA reductive dechlorination and reduced methanogenic activity. By contrast, hematite did not display any apparent stimulatory effect. Surprisingly, in the presence of higher amounts of iron oxides, a weaker effect was obtained, probably because iron(III) present in the minerals competed for the electrons necessary for reductive dechlorination. For all microcosms, the concentration of the toxic byproduct vinyl chloride was negligible throughout the whole study. The SEM/EDS analysis confirmed the close interaction between the conductive iron oxide particles and the dechlorinating bacteria. This work opens the possibility of using natural conductive minerals for bioremediation applications as well as shedding light on the previously unrecognized role of such minerals in contaminated ecosystems