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Alessia Pampuri, Alessio Tugnolo, Davide Bianchi, Valentina Giovenzana, Roberto Beghi, Natacha Fontes, Hugo M. Oliveira, Andrea Casson, Lucio Brancadoro, Riccardo Guidetti
Optical specifications for a proximal sensing approach to monitor the vine water status in a distributed and autonomous fashion Journal Article
BIOSYSTEMS ENGINEERING, 212 , pp. 388-398, 2021.
Abstract | Links | BibTeX
@article{Pampuri2021,
title = {Optical specifications for a proximal sensing approach to monitor the vine water status in a distributed and autonomous fashion},
author = {Alessia Pampuri, Alessio Tugnolo, Davide Bianchi, Valentina Giovenzana, Roberto Beghi, Natacha Fontes, Hugo M. Oliveira, Andrea Casson, Lucio Brancadoro, Riccardo Guidetti },
url = {https://doi.org/10.1016/j.biosystemseng.2021.11.007},
doi = {10.1016/j.biosystemseng.2021.11.007},
year = {2021},
date = {2021-12-16},
journal = {BIOSYSTEMS ENGINEERING},
volume = {212},
pages = {388-398},
abstract = {In agriculture, increasing attention is being paid to collect data in a non-destructive way using optical systems which can be field distributed in a completely interconnected network. To improve the irrigation scheduling management, the control of the plant's water status is crucial. This work focused on the definition of optical specifications (wavelength-selection in vis/NIR region) for the development of cost-effective sensors, giving an initial bulk of information to design optical devices to be used in a network of distributed field sensors. The analyses were performed on vines of cv. Pinot Blanc. Optical data were collected on leaves before the analysis of water potential and moisture content. Pearson-correlation analysis between predawn water potential (ѰPD) and moisture content was performed (r = 0.47 and p-value<0.05) highlighting a non-highly correlation between the two parameters. The optical data (350–2500 nm) were used to build a PLS-model with vis/NIR and ѰPD (RMSEP = 0.056 MPa, R2 = 0.7). The study identified the most significant wavelengths related to the water potential at the leaf level to design a chemometric model that was compared to the model based on the whole spectra. Therefore, related VIP-scores were used to calibrate another PLS-model after the selection of most relevant optical bands (530 ± 20 nm, 700 ± 20 nm, and 1400 ± 20 nm). Good predictive performance was obtained with an RMSEP = 0.056 and an R2 = 0.60. These results paved the ground for further development of integrated optical sensors capable to monitor vine water status in the field in a distributed and autonomous fashion.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
In agriculture, increasing attention is being paid to collect data in a non-destructive way using optical systems which can be field distributed in a completely interconnected network. To improve the irrigation scheduling management, the control of the plant's water status is crucial. This work focused on the definition of optical specifications (wavelength-selection in vis/NIR region) for the development of cost-effective sensors, giving an initial bulk of information to design optical devices to be used in a network of distributed field sensors. The analyses were performed on vines of cv. Pinot Blanc. Optical data were collected on leaves before the analysis of water potential and moisture content. Pearson-correlation analysis between predawn water potential (ѰPD) and moisture content was performed (r = 0.47 and p-value<0.05) highlighting a non-highly correlation between the two parameters. The optical data (350–2500 nm) were used to build a PLS-model with vis/NIR and ѰPD (RMSEP = 0.056 MPa, R2 = 0.7). The study identified the most significant wavelengths related to the water potential at the leaf level to design a chemometric model that was compared to the model based on the whole spectra. Therefore, related VIP-scores were used to calibrate another PLS-model after the selection of most relevant optical bands (530 ± 20 nm, 700 ± 20 nm, and 1400 ± 20 nm). Good predictive performance was obtained with an RMSEP = 0.056 and an R2 = 0.60. These results paved the ground for further development of integrated optical sensors capable to monitor vine water status in the field in a distributed and autonomous fashion.
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Catarina L. Nogueira, Diana P. Pires, Rodrigo Monteiro, Sílvio B. Santos, Carla M. Carvalho
Exploitation of a Klebsiella Bacteriophage Receptor-Binding Protein as a Superior Biorecognition Molecule Journal Article
ACS INFECTIOUS DISEASES, 7 (11), pp. 3077-3087, 2021.
Abstract | Links | BibTeX
@article{Nogueira2021b,
title = {Exploitation of a Klebsiella Bacteriophage Receptor-Binding Protein as a Superior Biorecognition Molecule},
author = {Catarina L. Nogueira, Diana P. Pires, Rodrigo Monteiro, Sílvio B. Santos, Carla M. Carvalho},
url = {https://doi.org/10.1021/acsinfecdis.1c00366},
doi = {10.1021/acsinfecdis.1c00366},
year = {2021},
date = {2021-11-12},
journal = {ACS INFECTIOUS DISEASES},
volume = {7},
number = {11},
pages = {3077-3087},
abstract = {Klebsiella pneumoniae is a Gram-negative bacterium that has become one of the leading causes of life-threatening healthcare-associated infections (HAIs), including pneumonia and sepsis. Moreover, due to its increasingly antibiotic resistance, K. pneumoniae has been declared a global top priority concern. The problem of K. pneumoniae infections is due, in part, to the inability to detect this pathogen rapidly and accurately and thus to treat patients within the early stages of infections. The success in bacterial detection is greatly dictated by the biorecognition molecule used, with the current diagnostic tools relying on expensive probes often lacking specificity and/or sensitivity. (Bacterio)phage receptor-binding proteins (RBPs) are responsible for the recognition and adsorption of phages to specific bacterial host receptors and thus present high potential as biorecognition molecules. In this study, we report the identification and characterization of a novel RBP from the K. pneumoniae phage KpnM6E1 that presents high specificity against the target bacteria and high sensitivity (80%) to recognize K. pneumoniae strains. Moreover, adsorption studies validated the role of gp86 in the attachment to bacterial receptors, as it highly inhibits (86%) phage adsorption to its Klebsiella host. Overall, in this study, we unravel the role and potential of a novel Klebsiella phage RBP as a powerful tool to be used coupled with analytical techniques or biosensing platforms for the diagnosis of K. pneumoniae infections.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Klebsiella pneumoniae is a Gram-negative bacterium that has become one of the leading causes of life-threatening healthcare-associated infections (HAIs), including pneumonia and sepsis. Moreover, due to its increasingly antibiotic resistance, K. pneumoniae has been declared a global top priority concern. The problem of K. pneumoniae infections is due, in part, to the inability to detect this pathogen rapidly and accurately and thus to treat patients within the early stages of infections. The success in bacterial detection is greatly dictated by the biorecognition molecule used, with the current diagnostic tools relying on expensive probes often lacking specificity and/or sensitivity. (Bacterio)phage receptor-binding proteins (RBPs) are responsible for the recognition and adsorption of phages to specific bacterial host receptors and thus present high potential as biorecognition molecules. In this study, we report the identification and characterization of a novel RBP from the K. pneumoniae phage KpnM6E1 that presents high specificity against the target bacteria and high sensitivity (80%) to recognize K. pneumoniae strains. Moreover, adsorption studies validated the role of gp86 in the attachment to bacterial receptors, as it highly inhibits (86%) phage adsorption to its Klebsiella host. Overall, in this study, we unravel the role and potential of a novel Klebsiella phage RBP as a powerful tool to be used coupled with analytical techniques or biosensing platforms for the diagnosis of K. pneumoniae infections.
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Ana T. Rufino, Ana Ramalho, Adelaide Sousa, José Miguel P. Ferreira de Oliveira, Paulo Freitas, Manuel A. Gonzalez Gómez, Yolanda Piñeiro-Redondo, José Rivas, Félix Carvalho, Eduarda Fernandes,* Marisa Freitas
Protective Role of Flavonoids against Intestinal Pro-Inflammatory Effects of Silver Nanoparticles Journal Article
MOLECULES, 26 (6610), 2021.
Abstract | Links | BibTeX
@article{Rufino2021,
title = {Protective Role of Flavonoids against Intestinal Pro-Inflammatory Effects of Silver Nanoparticles},
author = {Ana T. Rufino, Ana Ramalho, Adelaide Sousa, José Miguel P. Ferreira de Oliveira, Paulo Freitas, Manuel A. Gonzalez Gómez, Yolanda Piñeiro-Redondo, José Rivas, Félix Carvalho, Eduarda Fernandes,* Marisa Freitas},
url = {https://doi.org/10.3390/molecules26216610},
doi = {10.3390/molecules26216610},
year = {2021},
date = {2021-10-31},
journal = {MOLECULES},
volume = {26},
number = {6610},
abstract = {Silver nanoparticles (AgNP) have been increasingly incorporated into food-related and hygiene products for their unique antimicrobial and preservative properties. The consequent oral exposure may then result in unpredicted harmful effects in the gastrointestinal tract (GIT), which should be considered in the risk assessment and risk management of these materials. In the present study, the toxic effects of polyethyleneimine (PEI)-coated AgNP (4 and 19 nm) were evaluated in GIT-relevant cells (Caco-2 cell line as a model of human intestinal cells, and neutrophils as a model of the intestinal inflammatory response). This study also evaluated the putative protective action of dietary flavonoids against such harmful effects. The obtained results showed that AgNP of 4 and 19 nm effectively induced Caco-2 cell death by apoptosis with concomitant production of nitric oxide, irrespective of the size. It was also observed that AgNP induced human neutrophil oxidative burst. Interestingly, some flavonoids, namely quercetin and quercetagetin, prevented the deleterious effects of AgNP in both cell types. Overall, the data of the present study provide a first insight into the promising protective role of flavonoids against the potentially toxic effects of AgNP at the intestinal level.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Silver nanoparticles (AgNP) have been increasingly incorporated into food-related and hygiene products for their unique antimicrobial and preservative properties. The consequent oral exposure may then result in unpredicted harmful effects in the gastrointestinal tract (GIT), which should be considered in the risk assessment and risk management of these materials. In the present study, the toxic effects of polyethyleneimine (PEI)-coated AgNP (4 and 19 nm) were evaluated in GIT-relevant cells (Caco-2 cell line as a model of human intestinal cells, and neutrophils as a model of the intestinal inflammatory response). This study also evaluated the putative protective action of dietary flavonoids against such harmful effects. The obtained results showed that AgNP of 4 and 19 nm effectively induced Caco-2 cell death by apoptosis with concomitant production of nitric oxide, irrespective of the size. It was also observed that AgNP induced human neutrophil oxidative burst. Interestingly, some flavonoids, namely quercetin and quercetagetin, prevented the deleterious effects of AgNP in both cell types. Overall, the data of the present study provide a first insight into the promising protective role of flavonoids against the potentially toxic effects of AgNP at the intestinal level.
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Briliant Adhi Prabowo, Patrícia D. Cabral, Paulo Freitas, Elisabete Fernandes
The Challenges of Developing Biosensors for Clinical Assessment: A Review Journal Article
CHEMOSENSORS, 9 (11), pp. 299, 2021.
Abstract | Links | BibTeX
@article{Prabowo2021,
title = {The Challenges of Developing Biosensors for Clinical Assessment: A Review},
author = {Briliant Adhi Prabowo, Patrícia D. Cabral, Paulo Freitas, Elisabete Fernandes },
url = {https://doi.org/10.3390/chemosensors9110299},
doi = {10.3390/chemosensors9110299},
year = {2021},
date = {2021-10-24},
journal = {CHEMOSENSORS},
volume = {9},
number = {11},
pages = {299},
abstract = {Emerging research in biosensors has attracted much attention worldwide, particularly in response to the recent pandemic outbreak of coronavirus disease 2019 (COVID-19). Nevertheless, initiating research in biosensing applied to the diagnosis of diseases is still challenging for researchers, be it in the preferences of biosensor platforms, selection of biomarkers, detection strategies, or other aspects (e.g., cutoff values) to fulfill the clinical purpose. There are two sides to the development of a diagnostic tool: the biosensor development side and the clinical side. From the development side, the research engineers seek the typical characteristics of a biosensor: sensitivity, selectivity, linearity, stability, and reproducibility. On the other side are the physicians that expect a diagnostic tool that provides fast acquisition of patient information to obtain an early diagnosis or an efficient patient stratification, which consequently allows for making assertive and efficient clinical decisions. The development of diagnostic devices always involves assay developer researchers working as pivots to bridge both sides whose role is to find detection strategies suitable to the clinical needs by understanding (1) the intended use of the technology and its basic principle and (2) the preferable type of test: qualitative or quantitative, sample matrix challenges, biomarker(s) threshold (cutoff value), and if the system requires a mono- or multiplex assay format. This review highlights the challenges for the development of biosensors for clinical assessment and its broad application in multidisciplinary fields. This review paper highlights the following biosensor technologies: magnetoresistive (MR)-based, transistor-based, quartz crystal microbalance (QCM), and optical-based biosensors. Its working mechanisms are discussed with their pros and cons. The article also gives an overview of the most critical parameters that are optimized by developing a diagnostic tool.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Emerging research in biosensors has attracted much attention worldwide, particularly in response to the recent pandemic outbreak of coronavirus disease 2019 (COVID-19). Nevertheless, initiating research in biosensing applied to the diagnosis of diseases is still challenging for researchers, be it in the preferences of biosensor platforms, selection of biomarkers, detection strategies, or other aspects (e.g., cutoff values) to fulfill the clinical purpose. There are two sides to the development of a diagnostic tool: the biosensor development side and the clinical side. From the development side, the research engineers seek the typical characteristics of a biosensor: sensitivity, selectivity, linearity, stability, and reproducibility. On the other side are the physicians that expect a diagnostic tool that provides fast acquisition of patient information to obtain an early diagnosis or an efficient patient stratification, which consequently allows for making assertive and efficient clinical decisions. The development of diagnostic devices always involves assay developer researchers working as pivots to bridge both sides whose role is to find detection strategies suitable to the clinical needs by understanding (1) the intended use of the technology and its basic principle and (2) the preferable type of test: qualitative or quantitative, sample matrix challenges, biomarker(s) threshold (cutoff value), and if the system requires a mono- or multiplex assay format. This review highlights the challenges for the development of biosensors for clinical assessment and its broad application in multidisciplinary fields. This review paper highlights the following biosensor technologies: magnetoresistive (MR)-based, transistor-based, quartz crystal microbalance (QCM), and optical-based biosensors. Its working mechanisms are discussed with their pros and cons. The article also gives an overview of the most critical parameters that are optimized by developing a diagnostic tool.
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Coelho, Filipe; Botelho, C. M. ; Paris, Juan L.; Marques, Eduardo F.; Silva, Bruno F. B.
Influence of the media ionic strength on the formation and in vitro biological performance of polycation-DNA complexes Journal Article
JOURNAL OF MOLECULAR LIQUIDS, 344 (117930), 2021.
Abstract | Links | BibTeX
@article{Coelho2021,
title = {Influence of the media ionic strength on the formation and in vitro biological performance of polycation-DNA complexes},
author = {Coelho, Filipe; Botelho, C. M. ; Paris, Juan L.; Marques, Eduardo F.; Silva, Bruno F. B.},
url = {https://doi.org/10.1016/j.molliq.2021.117930},
doi = {10.1016/j.molliq.2021.117930},
year = {2021},
date = {2021-10-21},
journal = {JOURNAL OF MOLECULAR LIQUIDS},
volume = {344},
number = {117930},
abstract = {Cationic polymer-DNA complexes, or polyplexes, have been subject to intensive investigation as potentially efficient non-viral systems for gene therapy. Yet the effects of ionic strength, a physiologically relevant parameter, on the formation, physicochemical properties (e.g. size and colloidal stability) and transfection efficiency of polyplexes are still poorly investigated and understood. In this work, we analyze the effect of ionic strength on the formation and transfection efficiency of poly-L-lysine (PLL), branched polyethylenimine (PEI) and bioreducible poly-L-Lysine (bPLL) polycations complexed with plasmid DNA, using different preparation paths. In path I, the polycation and DNA are mixed in water and transferred to saline media afterwards. In path II, the polycation and DNA are mixed already in the presence of salt. Despite that the final compositions are identical, for monovalent salt (NaCl) concentrations ≥ 70 mM, the two pathways give rise to polyplexes with different sizes and stability. Path I polyplexes are smaller and colloidally more stable than their path II analogues, irrespective of polycation. Regarding the different polycations used, PLL-DNA polyplexes are smaller and more stable than PEI-polyplexes, regardless of path, while bPLL-DNA particles aggregate very easily in saline media. Conversely, when applied to 2D A549 cell cultures, the two assembly pathways do not show significant differences in transfection efficiency, but regarding cellular-uptake, PEI and path I offer better results. Overall, we show that slight differences in ionic strength at the time of polyplex formation strongly influence the size and stability of polycation-DNA complexes, but they do not translate into significant differences in the transfection of 2D in vitro A549 cell-cultures. Notwithstanding, caution should be exercised as the size differences observed could impact transfection in more complex in vivo models.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Cationic polymer-DNA complexes, or polyplexes, have been subject to intensive investigation as potentially efficient non-viral systems for gene therapy. Yet the effects of ionic strength, a physiologically relevant parameter, on the formation, physicochemical properties (e.g. size and colloidal stability) and transfection efficiency of polyplexes are still poorly investigated and understood. In this work, we analyze the effect of ionic strength on the formation and transfection efficiency of poly-L-lysine (PLL), branched polyethylenimine (PEI) and bioreducible poly-L-Lysine (bPLL) polycations complexed with plasmid DNA, using different preparation paths. In path I, the polycation and DNA are mixed in water and transferred to saline media afterwards. In path II, the polycation and DNA are mixed already in the presence of salt. Despite that the final compositions are identical, for monovalent salt (NaCl) concentrations ≥ 70 mM, the two pathways give rise to polyplexes with different sizes and stability. Path I polyplexes are smaller and colloidally more stable than their path II analogues, irrespective of polycation. Regarding the different polycations used, PLL-DNA polyplexes are smaller and more stable than PEI-polyplexes, regardless of path, while bPLL-DNA particles aggregate very easily in saline media. Conversely, when applied to 2D A549 cell cultures, the two assembly pathways do not show significant differences in transfection efficiency, but regarding cellular-uptake, PEI and path I offer better results. Overall, we show that slight differences in ionic strength at the time of polyplex formation strongly influence the size and stability of polycation-DNA complexes, but they do not translate into significant differences in the transfection of 2D in vitro A549 cell-cultures. Notwithstanding, caution should be exercised as the size differences observed could impact transfection in more complex in vivo models.
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Conference presentation: Fernandes E., Martins V., Dias T., Germano J., Sobrino T., Cardoso S., Castillo J., Rivas J., Freitas PP. Simultaneous interrogation of multiple targets using a magnetoresistive (MR) biochip platform. Biosensors 2016, Gothenburg, Sweden 25-27 May 2016
