Benchmarking of Nondestructive Testing for Additive Manufacturing - Valdemar R. Duarte; Tiago A. Rodrigues; Miguel A. Machado (3D Printing and Additive Manufacturing, 2021)
Defect detection in additive manufacturing (AM) is of paramount importance to improve the reliability of products. Nondestructive testing is not yet widely used for defect detection. The main challenges are a lack of standards and methods, the types and location of defects, and the complex geometry of many parts. During selective laser melting (SLM), several types of defects can occur such as porosity, cracking, and lack of fusion. In this study, several nondestructive tests were conducted in a highly complex shaped part in AISI 316L stainless steel with real defects manufactured by SLM. Two additional artificial defects (one horizontal and one flat bottom hole) were produced and the defect detectability was evaluated. The techniques used were as follows: dye penetrant, infrared thermography, immersion ultrasonic, eddy current, and X-ray microcomputed tomography to assess different types of defects in the as-built part. We conclude that no single technique can detect every type of defect, although multiple techniques provide complementary and redundant information to critically evaluate the integrity of the parts. This approach is fundamental for improving the reliability of defect detection, which will help expand the potential for using AM to produce parts for critical structural applications.

Bioelectricity generation using long-term operated biocathode: RFLP based microbial diversity analysis - S.V. Ramanaiaha; Cordas, C. M.; Matias, S.C.; M.V. Reddyd; J.H. Leitão; Fonseca, L. P. (Biotechnology Reports, 2021)

CARATERIZAÇÃO DE UMA MISTURA COMERCIAL DE UM MATERIAL CIMENTÍCIO DE ULTRAELEVADO DESEMPENHO - JOÃO FILIPE SOUSA PEREIRA (Faculty of Engineering of the University of Porto, 2021)

Carbon dioxide utilisation - bioelectrochemical approaches - Cordas C.M.; Moura J.J.G.; Escapa A.; Mateos R. (Enzymes for Solving Humankind's Problems, Moura J.J.G., Moura I., Maia L.B. (eds), 2021)

Carbon dioxide utilisation - the formate route - Maia L.B.; Moura I.; Moura J.J.G. (Enzymes for Solving Humankind's Problems, Moura J.J.G., Moura I., Maia L.B. (eds), 2021)

Cellulose acetate fibres loaded with daptomycin for metal implant coatings - Faria, Jaime; Bruno Dionísio; Iris Soares; Baptista, Ana Catarina; Ana Cláudia Marques; Lídia Gonçalves; Ana Bettencourt; Carlos Baleizão; Ferreira, Isabel (Carbohydrate polymers, 2021)
Multifunctional polymeric coatings containing drug delivery vehicles can play a key role in preventing/reducing biofilm formation on implant surfaces. Their requirements are biocompatibility, good adhesion, and controllable drug release. Although cellulose acetate (CA) films and membranes are widely studied for scaffolding, their applications as a protective coating and drug delivery vehicle for metal implants are scarce. The reason is that adhesion to stainless steel (SS) substrates is non-trivial. Grinding SS substrates enhances the adhesion of dip-coated CA films while the adhesion of electrospun CA membranes is improved by an electrosprayed chitosan intermediate layer. PMMA microcapsules containing daptomycin have been successfully incorporated into CA films and fibres. The released drug concentration of 3 x10-3 mg/mL after 120 minutes was confirmed from the peak luminescence intensity under UV radiation of simulated body fluid (SBF) after immersion of the fibres.

Characterization of CeO2 Nanoparticles Synthesized by the Precipitation in the Water-Alcohol Solutions - Shlapa, Yu.; Timashkov, I.; Veltruska, K.; Siposova, K.; Garcarova, I.; Musatov, A.; Solopan, S.; Kubovcikova, M.; Belous, A. (New Journal of Chemistry, 2021)

Colloidal Lithography for Photovoltaics: An Attractive Route for Light Management - Rui D. Oliveira; Mouquinho, Ana; Pedro Centeno; Alexandre, Miguel; Haque, Sirazul; Rodrigo Martins; Elvira Fortunato; Hugo Águas; Mendes, Manuel J. (Nanomaterials, 2021)

Colourful Patterns in Cellulose-Based Liquid Crystals - Silva, Pedro E. S.; da Rosa, Rafaela R.; Chagas, Ricardo; Fernandes, Susete N.; Godinho, Maria Helena (Liquid Crystals and their Application, 2021)

Colours of pre-cinema projections: the evolution of hand-painted magic lantern glass slides' palette - Ângela Santos; Otero, Vanessa; Vilarigues, Márcia (Proceedings of the International Colour Association (AIC) Conference 2021, 2021)

Complementary experimental and computational analysis of the effects of non-ionic detergents and phospholipids on insulin amyloid aggregation - Siposova,K., Kozar, T., Stupakova, M., Musatov, A. (Colloids and Surface B: Biointerfaces, 2021)

The complete catalytic mechanism of Xanthine Oxidase: a computational study - Fernandes, H.; Maia, L.; Ribeiro, P.M.; Moura J.J.G.; Cerqueira, N. M. (Inorg Chem Front, 2021)

Continuous wave terahertz imaging for NDT: Fundamentals and experimental validation - Frederico B. Costa; Miguel A. Machado; Grégoire J. Bonfait; Vieira, Pedro; Telmo G. Santos (Measurement, 2021)
Continuous wave terahertz (CW THz) imaging, is a variant of terahertz imaging that has been gaining scientific and technological relevance in multiple areas. In this paper the fundamental phenomena of CW THz were studied and a mathematical model was developed that successfully describes the Fabry–Perot interference for such a system, opening the possibility for measurement of thicknesses and surface curvatures. The capabilities of the system were tested using different types of defects, such as voids, water infiltrations and thin metallic wires. The interactions between different materials, features and the radiation beam were numerically studied using finite element method and the results agreed with the experiments. By comparing the results with other Non-Destructive Testing methods, it was found that CW THz imaging is particularly interesting to image water infiltrations and composite materials that incorporate conductive wires.

Cover image Advanced Material Journal - https://onlinelibrary.wiley.com/doi/abs/10.1002/adma.202170047 (2021)

"Defining the first preventive conservation guidelines for hand-painted magic lantern glass slides" - Ângela Santos; Beatriz Rodrigues; Otero, Vanessa; Vilarigues, Márcia (Conservar Património, 2021)

Design and engineering of magneto-responsive devices for cancer theranostics: Nano to macro perspective - Paula I P Soares; Joana Romão; Matos, Ricardo; Silva, Jorge Carvalho; Borges, João Paulo (Progress in Materials Science, 2021)
Design, research, and development of new and improved smart multifunctional devices is one of the main topics in the advanced functional materials agenda for the next decade. Smart materials that can be triggered by external stimuli are seen with high potential for innovative treatments and improved drug delivery systems by regulatory agencies like the FDA and EMA. The incorporation of magnetic nanostructures into complex systems produces multifunctional devices that can be spatiotemporally controlled by an external magnetic field. These magneto-responsive devices can be used for a multitude of biomedical applications, from diagnostic to the treatment of tumors, and are actively being developed and tested for cancer theranostics. Herein, we review the development of magneto-responsive devices for cancer theranostics, starting from the most straightforward architecture, single nanoparticles. We give some theoretical concepts about the design and production of such systems while providing a critical review of applications in clinical practice. Naturally, the review evolves to more complex architectures, from one-dimensional to three-dimensional magneto-responsive systems, demonstrating higher complexity and multifunctionality, and consequently, higher interest for clinical practice. The review ends with the main challenges in the design and engineering of magneto-responsive devices for cancer theranostics and future trends in this biomedical field.

Design and synthesis of low temperature printed metal oxide memristors - Carlos, E.; Deuermeier, J.; R. Branquinho; Gaspar, C; R. Martins; Kiazadeh, A.; E. Fortunato (Materials Chemistry C, 2021)

Development of an HPFRC for Use in Flat Slabs - Julia Blazy; Sandra Nunes; Carlos Sousa; Mário Pimentel (Fibre Reinforced Concrete: Improvements and Innovations. BEFIB 2020. RILEM Bookseries, vol 30., 2021)

Development of highly selective composite polymeric membranes for Li+/ Mg2+ separation - H.M. Saif; R.M. Huertas; S. Pawlowski; Crespo, J. G.; S. Velizarov (Journal of Membrane Science, 2021)

Development of Multifunctional Nanocomposites for Controlled Drug Delivery and Hyperthermia - Hovhannisyan, V.; Siposova, K.; Musatov, A.; Chen, Sh.-J. (Scientific Reports, 2021)

Dynamic Magnetic Response of Ferrofluids under a Static Electric Field - Rajnak, M.; Dolnik, B.; Hodermarsky, P.; Paulovicova, K.; Cimbala, R.; Timko, M.; Kopcansky, P. (Physics of Fluids, 2021)

eHooke: a tool for automated image analysis of spherical bacteria based on cell cycle progression - Saraiva, Bruno M; Krippahl, Ludwig; Filipe, Sergio R.; Henriques, Ricardo; Pinho, Mariana G. (Biological Imaging, 2021)

Electronic control of drug release from gauze or cellulose acetate fibres for dermal applications - Baptista, Ana Catarina; Brito, Miguel; Marques, Ana; Ferreira, Isabel (Journal of Materials Chemistry B, 2021)
Electronic controlled drug release from fibres was studied using ibuprofen as a model drug, one of the most popular analgesics, to impregnate gauze and cellulose acetate (CA) membranes. Conductivity in the range of 1–10 mS cm−1 was obtained in polypyrrole (Ppy) functionalised gauze and CA fibres, providing voltage-controlled drug release in a system consisting of Ppy/Ibuprofen/Ppy membranes and an Ag electrode. SEM images evidenced the Ppy adhesion to fibres and Micro Raman spectra proved drug incorporation and release. A small wound adhesive built with these membranes retains ibuprofen at 1.5 V and quickly releases it when −0.5 V is applied.

Electronic control of drug release from gauze or cellulose acetate fibres for dermal applications - Baptista, Ana Catarina; Brito, Miguel; Marques, Ana; Ferreira, Isabel (Journal of Materials Chemistry B, 2021)

Empowering Photovoltaics with Smart Light Management Technologies - Christian Stefano Schuster; Isodiana Crupi; Janne Halme; Mehmet Koç; Mendes, Manuel J.; Ian Marius Peters; Selçuk Yerci (Handbook of Climate Change Mitigation and Adaptation, 2021)

Fish gelatin-based films for gas sensing - Moreira, Inês Pimentel; Sato, Laura; Cláudia Alves; Susana Palma; Roque, Ana Cecília (BIODEVICES 2021 - 14th International Conference on Biomedical Electronics and Devices; Part of the 14th International Joint Conference on Biomedical Engineering Systems and Technologies, BIOSTEC 2021, 2021)
Electronic noses (e-noses) mimic the complex biological olfactory system, usually including an array of gas sensors to act as the olfactory receptors and a trained computer with signal-processing and pattern recognition tools as the brain. In this work, a new stimuli-responsive material is shown, consisting of self-assembled droplets of liquid crystal and ionic liquid stabilised within a fish gelatin matrix. These materials change their opto/electrical properties upon contact with volatile organic compounds (VOCs). By using an in-house developed e-nose, these new gas-sensing films yield characteristic optical signals for VOCs from different chemical classes. A support vector machine classifier was implemented based on 12 features of the signals. The results show that the films are excellent identifying hydrocarbon VOCs (toluene, heptane and hexane) (95% accuracy) but lower performance was found to other VOCs, resulting in an overall 60.4% accuracy. Even though they are not reusable, these sustainable gas-sensing films are stable throughout time and reproducible, opening several opportunities for future optoelectronic devices and artificial olfaction systems.
n/a
Keywords: Electronic nose; Fish gelatin; Gas-sensing; Ionic liquid; Liquid crystal; Volatile organic compounds

Flat Slabs with Different Longitudinal Reinforcement Ratios Under Horizontal Cyclic Loading - Mariana Rossi (NOVA School of Science and Technology, 2021)
The following dissertation studies the behavior of flat slabs when subjected to constant vertical loads and cyclic horizontal displacements, as a continuation of previous studies developed at FCT/UNL. The main focus of this research is to study the influence of flexural reinforcement on the seismic response of flat slabs. Therefore, three reinforced concrete flat slabs with varying flexural reinforcement ratio were tested, two having the same top reinforcement ratio of !=0,64% and one with !=1,34%. One of the specimens with lower longitudinal ratio was reinforced with studs as specific punching shear reinforcement. All slabs had overall dimensions of 4,15 × 1,85 × 0,15 m3 and a gravity shear ratio, ratio between the gravity load and the punching shear resistance, approximately equal to 55%. For a more complete analysis the results obtained were compared to two other specimens from previous experimental campaigns also conducted at FCT/UNL. These two slabs were designed with top flexural reinforcement ratio (!=0,96%) that lies between the two tested in this dissertation, one with no shear-reinforcement and the other with headed studs. Results showed that the reduction of flexural reinforcement resulted in a more ductile behavior of the specimens and in a higher drift capacity. The high flexural ratio added to one specimen improved the maximum unbalanced moment capacity but also made the slab fail in a more brittle mode. As expected, the specimen with shear headed studs supported the highest drifts and ended up not failing during this experimental campaign, reaching the test setup upper limit.

High-performance wide bandgap perovskite solar cells fabricated in ambient high-humidity conditions - Ugur Deneb Menda; Guilherme Ribeiro; Nunes, Daniela; Calmeiro, Tomás; Hugo Águas; Elvira Fortunato; Rodrigo Martins; Mendes, Manuel J. (Materials Advances, 2021)

Image Recording and Processing Chemical Synthesis: Method Description and Demonstration - Sousa, David M.; João Carlos Lima; Ferreira, Isabel (Chemistry‐Methods, 2021)

In Situ Electrochemical Characterization of a Microbial Fuel Cell Biocathode Running on Wastewater - S.V. Ramanaiah; Cordas, C. M.; S. Matiasand; Fonseca, L. P. (Catalysts, 2021)

Incorporation of Dual-Stimuli Responsive Microgels in Nanofibrous Membranes for Cancer Treatment by Magnetic Hyperthermia - Adriana Gonçalves; Filipe V. Almeida; Borges, João Paulo; Paula I P Soares (Gels, 2021)
The delivery of multiple anti-cancer agents holds great promise for better treatments. The present work focuses on developing multifunctional materials for simultaneous and local combinatory treatment: Chemotherapy and hyperthermia. We first produced hybrid microgels (MG), synthesized by surfactant-free emulsion polymerization, consisting of Poly (N-isopropyl acrylamide) (PNIPAAm), chitosan (40 wt.%), and iron oxide nanoparticles (NPs) (5 wt.%) as the inorganic component. PNIPAAm MGs with a hydrodynamic diameter of about 1 μm (in their swollen state) were successfully synthesized. With the incorporation of chitosan and NPs in PNIPAAm MG, a decrease in MG diameter and swelling capacity was observed, without affecting their thermosensitivity. We then sought to produce biocompatible and mechanically robust membranes containing these dual-responsive MG. To achieve this, MG were incorporated in poly (vinyl pyrrolidone) (PVP) fibers through colloidal electrospinning. The presence of NPs in MG decreases the membrane swelling ratio from 10 to values between 6 and 7, and increases the material stiffness, raising its Young modulus from 20 to 35 MPa. Furthermore, magnetic hyperthermia assay shows that PVP-MG-NP composites perform better than any other formulation, with a temperature variation of about 1 °C. The present work demonstrates the potential of using multifunctional colloidal membranes for magnetic hyperthermia and may in the future be used as an alternative treatment for cancer.

Influence of flexural reinforcement on the seismic performance of flat slab – column connections - Brisid Isufi; Mariana Rossi; António Pinho Ramos (Engineering Structures, 2021)
The behavior of flat slab – column connections under seismic-type loading is complex and not exhaustively studied. Among the many variables involved, this paper focuses on the influence of flexural reinforcement on the seismic performance of such connections. Three specimens were tested and analyzed in conjunction with two previously published specimens tested under similar conditions, under constant vertical loading and cyclic horizontal displacements, resulting in a total of five specimens. Among these specimens, the top flexural reinforcement varied from 0.64% to 1.34% and the approximate value of applied gravity shear ratio (GSR, equal to the ratio between the applied gravity load and the punching shear resistance) was around 55%. Two of the specimens (low and median reinforcement ratio) were also reinforced with headed studs against punching shear to study the unbalanced moment transfer capacity of the slab – column connections. The specimens are described and analyzed in detail. The results show that the performance under cyclic loading is affected by the amount of flexural reinforcement, even though GSR was almost the same for all specimens. It is shown that current code-based approaches for the estimation of unbalanced moment capacity, as well as drift capacity, are generally safe sided for the specimens under investigation but do not fully capture the trends observed in the experimental campaign.

Injectable Composite Systems Based on Microparticles in Hydrogels for Bioactive Cargo Controlled Delivery - Henrique Carrêlo; Paula I P Soares; Borges, João Borges; Maria Teresa Cidade (Gels, 2021)
Engineering drug delivery systems (DDS) aim to release bioactive cargo to a specific site within the human body safely and efficiently. Hydrogels have been used as delivery matrices in different studies due to their biocompatibility, biodegradability, and versatility in biomedical purposes. Microparticles have also been used as drug delivery systems for similar reasons. The combination of microparticles and hydrogels in a composite system has been the topic of many research works. These composite systems can be injected in loco as DDS. The hydrogel will serve as a barrier to protect the particles and retard the release of any bioactive cargo within the particles. Additionally, these systems allow different release profiles, where different loads can be released sequentially, thus allowing a synergistic treatment. The reported advantages from several studies of these systems can be of great use in biomedicine for the development of more effective DDS. This review will focus on in situ injectable microparticles in hydrogel composite DDS for biomedical purposes, where a compilation of different studies will be analysed and reported herein.

Injectable hydrogels with two different rates of drug release based on pluronic/water system filled with poly(ε-caprolactone) microcapsules - Beatriz Delgado; Henrique Carrêlo; Mónica V. Loureiro; Ana C. Marques; Borges, João Borges; Maria Teresa Cidade (Journal of Materials Science, 2021)
The present paper regards the preparation and characterization of Pluronic F127 + F68/water/poly (ε-caprolactone) microcapsules (MCs) composite systems for tissue repair. The first part of the work relates to the production of poly(ε-caprolactone) (PCL) MCs via water-in-oil-in-water (W/O/W) double emulsion system combined with solvent evaporation method. The study of different process parameters in the final MCs characteristics and their drug release profile is herein reported. Different percentages of PCL, emulsion stabilizer, and volume proportions of the emulsion constituents have been tested, leading to considerable differences in the MCs size distributions. The selected MCs, containing an aqueous solution of methylene blue (MB) as a model drug, were then used to fill a Pluronic F127 + F68/water system leading to the final composite system (5 and 10 wt % MB loaded PCL MCs). The composite systems were characterised in the second part of the work in terms of its rheological behaviour and drug release performance. They were found to gellify at 30 °C, and present an extended drug release to a total of 18 days. The models that best define the release profiles were also studied, with the release of MB occurring mostly by Fick diffusion and polymer chain relaxation. Pluronic F127 + F68/water/poly (ε-caprolactone) MCs composite system is shown to be a promising injectable system, with two different drug release rates, for tissue repair.

Lazy State Determination for SQL Databases - Eduardo Subtil (FCT-NOVA, 2021)

Light management with quantum nanostructured dots-in-host semiconductors - Alexandre, Miguel; Hugo Águas; Elvira Fortunato; Rodrigo Martins; Mendes, Manuel J. (Light: Science & Applications, 2021)

Low-cost and high-performance 3D printed YBCO superconductors - Diogo Mendes; David Sousa; Ana C Cerdeira; Laura C. J. Pereira; Marques, Ana; João Murta-Pina; Anabela Pronto; Ferreira, Isabel (Ceramics International, 2021)

Magnetic particles used in a new approach for designed protein crystallization - R. Santos; Romão, M.J.; Roque, A. C. A.; Carvalho, A. L. (CrystEngComm, 2021)

Mechanical and Fracture Behaviour of an HPFRC - Sandra Nunes; Mário Pimentel; Carlos Sousa (Proceedings of RILEM-fib International Symposium on Fibre Reinforced Concrete, BEFIB2021, 2021)
The current paper analyses the mechanical and fracture behaviour of a High-Performance Fibre Reinforced Concrete (HPFRC). An HPFRC was developed in a previous stage aiming to simultaneously, maximise aggregates content, achieve a compressive strength of 90–120 MPa and maintaining self-compactability (SF1+VS2). The benefits of fibres hybridisation (using fibres with lengths of 13, 35 and 60 mm) on flexural strength are investigated using the wedge-splitting test, in order to achieve the highest performance while keeping a relatively low fibre content. The final selected mixture was characterised in terms of workability, compressive strength and modulus of elasticity. Six notched prismatic specimens were subjected to three-point bending tests, according to EN 14651, for classification according to the MC2010. Based on the bending tests data, the simplified linear characteristic tensile stress vs. crack opening displacement relationship of the HPFRC was evaluated according to MC2010 and two other analytical approaches available in the literature.

A Molecular Link between Cell Wall Biosynthesis, Translation Fidelity, and Stringent Response in Streptococcus pneumoniae - Aggarwal, S D; Lloyd*, A J; Yerneni, S S; A. R. Narciso; Shepherd, J; Roper, D I; Dowson, C; S. R. Filipe*; Hiller*, N L (Proc. Natl. Acad. Sci. USA, 2021)

Monitoring the Surface Tension by the Pendant Drop Technique for Detection of Insulin Fibrillogenesis - Siposova, K.; Sedlakova, D.; Musatov, A. (Analytical Methods, 2021)

The Mycobacteriophage Ms6 LysB N-terminus Displays Peptidoglycan Binding Affinity - Gigante, A M; Olivença, F; M. J. Catalão; Moniz-Pereira, J; S. R. Filipe; M. Pimentel (Viruses, 2021)

Nanomaterials for magnetic hyperthermia - Manuel P. Graça; Silvia Soreto Teixeira; Sílvia R. Gavinho; Manuel Almeida Valente; Cristiana Salgueiro; José Nunes; Paula I P Soares; Lança, Maria Carmo; Tânia Vieira; Silva, Jorge Carvalho; Borges, João Borges (European Journal of Public Health, 2021)
Cancer remains as one of the major causes of mortality worldwide. Recent advances in nanoparticles based therapy mark a new era on cancer treatment. Many groups have investigated biological/physical effects of nanoparticles on tumour cells and how these vary with physical parameters such as particle size, shape, concentration and distribution. Magnetic hyperthermia (MHT) can be an alternative or an add-value therapy with demonstrated effectiveness. MHT uses magnetic nanoparticles, which can be directly applied to the tumour, where, by applying an external ac magnetic field, will promote a localized temperature increment that can be controlled.

Nanostructured LiFe5O8 by a Biogenic Method for Applications from Electronics to Medicine - Silvia Soreto Teixeira; Manuel P. F. Graça; José Lucas; Manuel Almeida Valente; Paula I P Soares; Lança, Maria Carmo; Tânia Vieira; Silva, Jorge Carvalho; Borges, João Paulo; Luiza-Izabela Jinga; Gabriel Socol; Cristiane Mello Salgueiro; et al. (Nanomaterials, 2021)
The physical properties of the cubic and ferrimagnetic spinel ferrite LiFe5O8 has made it an attractive material for electronic and medical applications. In this work, LiFe5O8 nanosized crystallites were synthesized by a novel and eco-friendly sol-gel process, by using powder coconut water as a mediated reaction medium. The dried powders were heat-treated (HT) at temperatures between 400 and 1000 °C, and their structure, morphology, electrical and magnetic characteristics, cytotoxicity, and magnetic hyperthermia assays were performed. The heat treatment of the LiFe5O8 powder tunes the crystallite sizes between 50 nm and 200 nm. When increasing the temperature of the HT, secondary phases start to form. The dielectric analysis revealed, at 300 K and 10 kHz, an increase of ε′ (≈10 up to ≈14) with a tanδ almost constant (≈0.3) with the increase of the HT temperature. The cytotoxicity results reveal, for concentrations below 2.5 mg/mL, that all samples have a non-cytotoxicity property. The sample heat-treated at 1000 °C, which revealed hysteresis and magnetic saturation of 73 emu g−1 at 300 K, showed a heating profile adequate for magnetic hyperthermia applications, showing the potential for biomedical applications.

Near infrared photothermoelectric effect in transparent AZO/ITO/Ag/ITO thin films - Bianchi, Catarina; Marques, AC; R.C. da Silva; Calmeiro, Tomas; Ferreira, Isabel (Scientific reports, 2021)

A Neural Network Approach towards Generalized Resistive Switching Modelling - Carvalho, Guilherme; Pereira, Maria; Kiazadeh, Asal; Tavares, Vitor Grade (Micromachines, 2021)

A New Long-Term Composite Drug Delivery System Based on Thermo-Responsive Hydrogel and Nanoclay - Cezar Tipa; Maria Teresa Cidade; Tânia Vieira; Silva, Jorge Carvalho; Paula I P Soares; Borges, João Paulo (Nanomaterials, 2021)
Several problems and limitations faced in the treatment of many diseases can be overcome by using controlled drug delivery systems (DDS), where the active compound is transported to the target site, minimizing undesirable side effects. In situ-forming hydrogels that can be injected as viscous liquids and jellify under physiological conditions and biocompatible clay nanoparticles have been used in DDS development. In this work, polymer–clay composites based on Pluronics (F127 and F68) and nanoclays were developed, aiming at a biocompatible and injectable system for long-term controlled delivery of methylene blue (MB) as a model drug. MB release from the systems produced was carried out at 37 °C in a pH 7.4 medium. The Pluronic formulation selected (F127/F68 18/2 wt.%) displayed a sol/gel transition at approx. 30 °C, needing a 2.5 N force to be injected at 25 °C. The addition of 2 wt.% of Na116 clay decreased the sol/gel transition to 28 °C and significantly enhanced its viscoelastic modulus. The most suitable DDS for long-term application was the Na116-MB hybrid from which, after 15 days, only 3% of the encapsulated MB was released. The system developed in this work proved to be injectable, with a long-term drug delivery profile up to 45 days.

Novo desenho das células dos painéis solares vai revolucionar o aproveitamento da luz solar - Mendes, Manuel J. (Agir pelo Planeta, 2021)

Optical Gas Sensing with Liquid Crystal Droplets and Convolutional Neural Networks - José Frazão; Susana I C J Palma; Henrique M.A. Costa; Alves, C.; Roque, Ana C A; Silveira, M (Sensors, 2021)

An Overview of High-Entropy Alloys as Biomaterials - Diogo Castro; Pedro Jaeger; Baptista, Ana Catarina; João Pedro Oliveira (Metals, 2021)
High-entropy alloys (HEAs) have been around since 2004. The breakthroughs in this field led to several potential applications of these alloys as refractory, structural, functional, and biomedical materials. In this work, a short overview on the concept of high-entropy alloys is provided, as well as the theoretical design approach. The special focus of this review concerns one novel class of these alloys: biomedical high-entropy alloys. Here, a literature review on the potential high-entropy alloys for biomedical applications is presented. The characteristics that are required for these alloys to be used in biomedical-oriented applications, namely their mechanical and biocompatibility properties, are discussed and compared to commercially available Ti6Al4V. Different processing routes are also discussed.

An Overview of High-Entropy Alloys as Biomaterials - D. Castro; P. Jaeger; Baptista, A. C.; Oliveira, J.P. (Metals, 2021)

Photonic Strategies for Photovoltaics: New Advances Beyond Optics - Mouquinho, Ana; Sanchez-Sobrado, Olalla; Haque, Sirazul; Pedro Centeno; Alexandre, Miguel F.; Guilherme Ribeiro; Jenny L. N. Boane; Mateus, Tiago; Ugur Deneb Menda; Hugo Águas; Elvira Fortunato; Rodrigo Martins; et al. (Modern Environmental Science and Engineering, 2021)

A purification platform for antibodies and derived fragments using a de novo designed affinity adsorbent - Matos, MJB; Trovão, F; Gonçalves, J.; Rothbauer, U; Freire, MG; Barbosa, AMJB; Pina, A. S.; Roque, A. C. A. (Separation and Purification Technology, 2021)

Real-time droplet monitoring for digital Polymerase Chain Reaction in microfluidic chip - Mota, Ana Catarina Candeias (Faculdade de Ciências e Tecnologia, 2021)
Current cancer diagnosis techniques are often dependent on the collection of tumour tissue, involving invasive processes for the patient. Circulating Tumour DNA (ctDNA) emerges as an alternative resource for cancer detection and monitoring, that can be har vested from simple blood samples. Digital Polymerase Chain Reaction (dPCR) is a fast and sensitive technique for DNA amplification, suitable for low DNA concentrations such as ctDNA. Advances in microfluidics allow the partition of PCR samples into droplets based in water-in-oil emulsions, so that PCR amplification occurs within each droplet. In this way, the PCR reaction is a well controlled process with a low probability of contami nation and allowing a high throughput analysis. The aimed of this work was to develop droplet-based microfluidic device for application to dPCR technique coupled with real-time droplet monitoring. This work focused on the design and fabrication of a microfluidic device capable of producing a large number of uniform droplets with volumes in the nanoliter range and constant frequency. For this, a polydimethylsiloxane (PDMS) droplet generator device was developed, through photo and soft-lithography techniques, and tested with several oil/water flow rates ratios. Then, the droplets generated were characterized in terms of droplet size, velocity and frequency through the implementation of a powerful open-source software for real-time analysis. Several tests on different devices were carried out to evaluate the device reproducibility. Finally, the droplet generator was incorporated with a serpentine design, allowing the PCR cycles to occur in continuous flow. The results revealed that was possible to generate droplets with radius between 22-99 µm and a coefficient of variation bellow 10%. The correspondents volumes ranged between 90 pL-4.18 nL. Moreover, the velocities obtained situated between 0.05 mm/s-7.62 mm/s with droplet generating frequency of 2-50 Hz. Regarding to the droplet monitoring, the results of the workflows developed revealed similarity with the results obtained trough a widely used software for this purposes, with the advantage of allowing real-time analysis for a larger sample of results.

Recent automation trends in Portugal: implications on industrial productivity and employment in automotive sector - Nuno Boavida; Candeias, Marta (Societies, 2021)

Recent Automation Trends in Portugal: Implications on Industrial Productivity and Employment in Automotive Sector - Nuno Boavida; Candeias, Marta (Societies, 2021)

Recent Progress in Solution‐Based Metal Oxide Resistive Switching Devices - Carlos, E.; Branquinha, R; R. Martins; Kiazadeh, A.; E. Fortunato (Advanced Material, 2021)

Remote Monitoring Systems for Patients With Chronic Diseases in Primary Health Care: Systematic Review - Mariana Peyroteo; Inês Augusto Ferreira; Luís Brito Elvas; João Carlos Ferreira; Luís Velez Lapão (JMIR Mhealth and Uhealth, 2021)

A review of tests on slab-column connections with advanced concrete materials - Brisid Isufi; António Pinho Ramos (Structures, 2021)
Advances in concrete technology during the last decades have resulted in the development of materials with enhanced mechanical properties, such as High Strength Concrete (HSC), Fibre Reinforced Concrete (FRC) and Ultra-High Performance Fibre Reinforced Concrete (UHPFRC). The application of these materials in flat slabs, which are a popular structural solution in Reinforced Concrete (RC) buildings worldwide, has the potential of significantly reducing raw material consumption by enabling the design of slenderer and therefore lighter structures. However, flat slabs are susceptible to punching shear failure, which is a complex phenomenon that remains challenging, even though significant efforts have been made to experimentally study it. For advanced concrete materials (HSC, FRC and UHPFRC), the challenge is further accentuated by the continuous and rapid development of these materials. With the purpose of identifying and highlighting gaps in the published literature, a review of tests with HSC, FRC and UHPFRC slab-column connections in non-seismic and seismic loading applications is presented in this paper. It is shown that future research directions in this field include, among others, testing thicker slabs, HSC slabs with higher concrete compressive strength, HSC combined with FRC and several more cases related to seismic loading conditions.

Screening of Potential Stress Biomarkers in Sweat Associated with Sports Training - Nunes, M. J.; Cordas, C. M.; Moura, J. J. G.; J. P. Noronha; Branco, L. C. (Sports Medicine - Open, 2021)

Social Dimensions in CPS & IoT Based Automated Production Systems - El-Haouzi, Hind B.; Valette, E.; Krings, B.-J.; Moniz, A. B. (Societies, 2021)

Soft-Microstructured Transparent Electrodes for Photonic-Enhanced Flexible Solar Cells - Jenny L. N. Boane; Pedro Centeno; Mouquinho, Ana; Alexandre, Miguel; Calmeiro, Tomás; Elvira Fortunato; Rodrigo Martins; Mendes, Manuel J.; Hugo Águas (Micro, 2021)

Stable and Oriented Liquid Crystal Droplets Stabilized by Imidazolium Ionic Liquids - Ramou, E.; Rebordao, G; S. I. C. J. Palma; Roque, A. C. A. (MOLECULES, 2021)

STRENGTHENING OF REINFORCED CONCRETE ELEMENTS WITH UHPFRC - Aurélio Guilherme Sine (Faculty of Engineering of the University of Porto, 2021)

Structural and Physical-Chemical Characterization of Redox Active CeO2 Nanoparticles Synthesized by the Precipitation in the Water-Alcohol Solutions - Shlapa, Yu; Timashkov, I.; Veltruska, K.; Siposova, K.; Garcarova, I.; Musatov, A.; Solopan, S.; Kubovcikova, M.; Belous, A. (Nanotechnology, 2021)

Technology as enabler of the automation of work? Current societal challenges for a future perspective of work - Moniz, A. B.; Krings, B.-J.; Frey, P. (Revista Brasileira de Sociologia, 2021)

Towards Sustainable Crossbar Artificial Synapses with Zinc-Tin Oxide - Silva, C; Martins, J.; Deuermeier, J.; Pereira, M.; Rovisco, A.; P. Barquinha; Goes, J.; E. Fortunato; Martins. R; Kiazadeh, A. (Electronics Material, 2021)

Cation-mediated gelation of the fucose-rich polysaccharide FucoPol: preparation and characterization of hydrogel beads and their cytotoxicity assessment - Fialho, Letícia; Araújo, Diana; Alves, Vitor D.; Catarina Roma-Rodrigues; Baptista, Pedro V; Fernandes, Alexandra R; Freitas, Filomena; Reis, Maria A. M. (International Journal of Polymeric Materials and Polymeric Biomaterials, 2021)
This study describes for the first time the iron- and copper-mediated gelation of FucoPol, fucose-rich bacterial polysaccharide. The ability of FucoPol to gel in the presence of metal cations, including iron(III) and copper(II), was used for the preparation of hydrogel beads. Iron mediated the formation of stable and not cytotoxic gel beads, while copper resulted in fragile and cytotoxic ones. Copper-mediated beads coated with an iron-mediated gel layer were more stable and had reduced cytotoxicity. The resulting polymeric structures had differing morphology, physical properties and cytotoxicity, which support their use in several applications, including biomedicine, agriculture and bioremediation.
FCT/MEC (UID/Multi/04378/2019). PEst-OE/AGR/UI0245/2014. POCI-01-0145-FEDER-007728 and POCI-01-0145-FEDER - 007265. SFRH/BD/140829/2018. SFRH/BPD/124612/2016.
Keywords: cytotoxicity; FucoPol; hydrogel beads

Donor Functionalized Iron(II) N-Heterocyclic Carbene Complexes in Transfer Hydrogenation Reactions - Lopes, Rita; Raya-Barón, Álvaro; Robalo, M. Paula; Vinagreiro, Carolina; Barroso, Sónia; Romão, Maria J.; Fernández, Ignacio; Pereira, Mariette M.; Royo, Beatriz (European Journal of Inorganic Chemistry, 2021)
Two piano-stool iron(II) complexes bearing N-heterocyclic carbene ligands outfitted with acetamide- and amine-pendant arms [Cp*Fe(NHCR)(CO)I] {Cp* = η5-tetramethylcyclopentadienyl; R = CH2CONEt2 (3), (CH2)2NEt2 (4)}, have been prepared and fully characterized. Their catalytic activity in transfer hydrogenation (TH) of ketones using iPrOH as a hydrogen source and catalytic amounts of base (LiOtBu) has been explored, along with that of previously reported [CpFe(NHCR)(CO)I] {R = nBu (5), (CH2)2OH (6), Et (7), and (CH2)3OH (8)} complexes containing hydroxyl and nonfunctionalized alkyl arms. Complex 3 displayed the highest catalytic activity of the whole series 3–8, reaching a TOF50 value of 533 h–1. NMR monitoring of the stoichiometric reaction of 3 with LiOtBu, allowed the identification of a new species 3' containing a deprotonated amidate moiety, which has been fully characterized by 1H, 13C, and 15N NMR. Finally, a green protocol for the reduction of ketones through TH using glycerol as a hydrogen source, under microwave irradiation in the presence of catalytic amounts of 3 and base has been developed.
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Keywords: Carbene ligands; Iron; Ketones; Microwave irradiation; Reduction; Transfer hydrogenation

A dual cohesin–dockerin complex binding mode in Bacteroides cellulosolvens contributes to the size and complexity of its cellulosome - Duarte, Marlene; Viegas, Aldino; Alves, Victor D.; Prates, José A. M.; Ferreira, Luís M. A.; Najmudin, Shabir; Cabrita, Eurico J.; Ana Luísa Carvalho; Fontes, Carlos M. G. A.; Bule, Pedro (Journal of Biological Chemistry, 2021)
The Cellulosome is an intricate macromolecular protein complex that centralizes the cellulolytic efforts of many anaerobic microorganisms through the promotion of enzyme synergy and protein stability. The assembly of numerous carbohydrate processing enzymes into a macromolecular multiprotein structure results from the interaction of enzyme-borne dockerin modules with repeated cohesin modules present in noncatalytic scaffold proteins, termed scaffoldins. Cohesin–dockerin (Coh-Doc) modules are typically classified into different types, depending on structural conformation and cellulosome role. Thus, type I Coh-Doc complexes are usually responsible for enzyme integration into the cellulosome, while type II Coh-Doc complexes tether the cellulosome to the bacterial wall. In contrast to other known cellulosomes, cohesin types from Bacteroides cellulosolvens, a cellulosome-producing bacterium capable of utilizing cellulose and cellobiose as carbon sources, are reversed for all scaffoldins, i.e., the type II cohesins are located on the enzyme-integrating primary scaffoldin, whereas the type I cohesins are located on the anchoring scaffoldins. It has been previously shown that type I B. cellulosolvens interactions possess a dual-binding mode that adds flexibility to scaffoldin assembly. Herein, we report the structural mechanism of enzyme recruitment into B. cellulosolvens cellulosome and the identification of the molecular determinants of its type II cohesin–dockerin interactions. The results indicate that, unlike other type II complexes, these possess a dual-binding mode of interaction, akin to type I complexes. Therefore, the plasticity of dual-binding mode interactions seems to play a pivotal role in the assembly of B. cellulosolvens cellulosome, which is consistent with its unmatched complexity and size.
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Keywords: cellulase; Cellulose; cellulosome; cohesin; Crystal structure; dockerin; dual-binding; protein complex

Enhanced Stability of Detergent-Free Human Native STEAP1 Protein from Neoplastic Prostate Cancer Cells upon an Innovative Isolation Procedure - Barroca-Ferreira, Jorge; Cruz-Vicente, Pedro; Santos, Marino F. A.; Rocha, Sandra M.; Santos-Silva, Teresa; Maia, Cláudio J.; Passarinha, Luís A. (International Journal of Molecular Sciences, 2021)
Background: The STEAP1 is a cell-surface antigen over-expressed in prostate cancer, which contributes to tumor progression and aggressiveness. However, the molecular mechanisms underlying STEAP1 and its structural determinants remain elusive. Methods: The fraction capacity of Butyl- and Octyl-Sepharose matrices on LNCaP lysates was evaluated by manipulating the ionic strength of binding and elution phases, followed by a Co-Immunoprecipitation (Co-IP) polishing. Several potential stabilizing additives were assessed, and the melting temperature (Tm) values ranked the best/worst compounds. The secondary structure of STEAP1 was identified by circular dichroism. Results: The STEAP1 was not fully captured with 1.375 M (Butyl), in contrast with interfering heterologous proteins, which were strongly retained and mostly eluted with water. This single step demonstrated higher selectivity of Butyl-Sepharose for host impurities removal from injected crude samples. Co-IP allowed recovering a purified fraction of STEAP1 and contributed to unveil potential physiologically interacting counterparts with the target. A Tm of  55 °C was determined, confirming STEAP1 stability in the purification buffer. A predominant α-helical structure was identified, ensuring the protein’s structural stability. Conclusions: A method for successfully isolating human STEAP1 from LNCaP cells was provided, avoiding the use of detergents to achieve stability, even outside a membrane-mimicking environment.
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Ibuprofen incorporated into unmodified and modified mesoporous silica: From matrix synthesis to drug release - Sara Inocêncio; Cordeiro, Teresa; Matos, Inês; Florence Danède,; Sotomayor, João C.; Fonseca, Isabel M.; Natália T. Correia; Corvo, Marta C.; Madalena Dionísio (Microporous and Mesoporous Materials, 2021)
Aiming to rationalize the release profile of an incorporated pharmaceutical drug in terms of its mobility, driven by guest-host interactions, the poorly water-soluble ibuprofen drug was loaded in a mesoporous inorganic silica matrix with unmodified (MCM-41) and modified surface (MCM-41sil) by post-synthesis silylation, both having pore sizes   3 nm. The single calorimetric detection of a broad glass transition step for both ibuprofen composites indicates full drug amorphization, confirmed by the only appearance of an amorphous halo in the powder XRD patterns. Moreover, a gradient profile is disclosed by the heat flux derivative plot in the glass transition, in coherence with the thermogravimetric profile that shows a multi-step decomposition trace for confined ibuprofen in these matrixes. While identical guest dynamics, as probed by dielectric relaxation spectroscopy, were found in both dehydrated composites, a significant molecular population with faster relaxation exists in the hydrated state for the drug inside the unmodified matrix. This was rationalized as the concurrence of true confinement effects, which manifest under nanometer dimensions, and greater water affinity of the unmodified matrix, forcing the drug molecules to be placed mostly in the pore core. Finite size effects are also felt in both dehydrated composites, however guest-host interactions give origin to a dominant population with slowed down mobility that governs the overall guest dynamics. In spite of an inferior number of active sites for drug adsorption in the silylated matrix, a faster ibuprofen delivery in phosphate buffer (pH = 6.8) was observed when the drug is released from unmodified MCM-41 in the hydrated state. Therefore, our results suggest that a relevant role is played by water molecules, which impair a strong guest adsorption in the host surface more efficiently than the limited surface modification, influence the higher ratio of a faster population in the pore core and facilitate the diffusion of the aqueous releasing media inside pores.
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Keywords: Amorphous materials; Confinement; Dielectric spectroscopy; Drug delivery; Magic angle spinning nuclear magnetic spectroscopy

Isothermal Amplification of Nucleic Acids: The Race for the Next “Gold Standard” - Oliveira, Beatriz B.; Bruno Veigas; Baptista, Pedro Viana (Frontiers in Sensors, 2021)
Nucleic acid amplification technologies (NAATs) have become fundamental tools in molecular diagnostics, due to their ability to detect small amounts of target molecules. Since its development, Polymerase Chain Reaction (PCR) has been the most exploited method, being stablished as the “gold standard” technique for DNA amplification. However, the requirement for different working temperatures leads to the need of a thermocycler machine or complex thermal apparatus, which have been preventing its application in novel integrated devices for single workflow and high throughput analysis. Conversely, isothermal amplification methods have been gaining attention, especially for point-of-care diagnosis and applications. These non-PCR based methods have been developed by mimicking the in vivo amplification mechanisms, while performing the amplification with high sensitivity, selectivity and allowing for high-throughput analysis. These favorable capabilities have pushed forward the implementation and commercialization of several platforms that exploit isothermal amplification methods, mostly against virus, bacteria and other pathogens in water, food, environmental and clinical samples. Nevertheless, the future of isothermal amplification methods is still dependent on achieving technical maturity and broader commercialization of enzymes and reagents.
Funding Information: info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F04378%2F2020/PT# info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F04378%2F2020/PT# info:eu-repo/grantAgreement/FCT/Concurso para Atribuição do Estatuto e Financiamento de Laboratórios Associados (LA)/LA%2FP%2F0140%2F2020/PT# info:eu-repo/grantAgreement/FCT//2020.07660.BD/PT# Publisher Copyright: Copyright © 2021 Oliveira, Veigas and Baptista.
Keywords: Isothermal amplification; NAAT; non-pcr amplification; nucleic acids amplification; point—of—care

Magnetic particles used in a new approach for designed protein crystallization - Raquel dos Santos; Romão, Maria João; Roque, Ana Cecília A; Ana Luísa Carvalho (CrystEngComm, 2021)
After more than one hundred and thirty thousand protein structures determined by X-ray crystallography{,} the challenge of protein crystallization for 3D structure determination remains. In the quest for additives for efficient protein crystallization{,} inorganic materials emerge as an alternative. Magnetic particles (MPs) are versatile inorganic materials{,} easy to use{,} modify and manipulate in a wide range of biological assays. The potential of using functionalised MPs as crystallization chaperones for protein crystallization was shown in this work. MPs with distinct coatings were rationally designed to promote protein crystallization by affinity-triggered heterogeneous nucleation. Hen egg white lysozyme (HEWL) and trypsin{,} were crystallized in the presence of MPs either bare or coated with a polysaccharide (chitin) or a protein (casein){,} respectively. The addition of MPs was characterized in terms of bound protein to the MPs{,} crystal morphology{,} time-lapse of crystal emergence{,} crystallization yield fold change and crystal diffraction quality for structure determination. The MPs additives have shown to bind to the respective target protein{,} and to promote nucleation and crystal growth without compromising crystal morphology. On the other hand{,} MPs addition led to faster detectable crystal emergence and up to 13 times higher crystallization yield{,} addressing some the challenges in protein crystallization{,} the main bottleneck of macromolecular crystallography. Structure determination of the protein crystallized in the presence of MPs revealed that the structural characteristics of the protein remained unchanged{,} as shown by the superposition with PDB annotated proteins. Moreover{,} and unlike most reported cases{,} it was possible to exclude the inhibitor benzamidine during trypsin crystallisation{,} which is a remarkable result opening new prospects in enzyme engineering and drug design. Our results show that MPs coated with affinity ligands to target proteins can be used as controlled and tailor-made crystallization inducers.
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Multistep Signaling in Nature: A Close-Up of Geobacter Chemotaxis Sensing - Silva, , Marta A.; Salgueiro, , Carlos A. (International Journal of Molecular Sciences, 2021)
Environmental changes trigger the continuous adaptation of bacteria to ensure their survival. This is possible through a variety of signal transduction pathways involving chemoreceptors known as methyl-accepting chemotaxis proteins (MCP) that allow the microorganisms to redirect their mobility towards favorable environments. MCP are two-component regulatory (or signal transduction) systems (TCS) formed by a sensor and a response regulator domain. These domains synchronize transient protein phosphorylation and dephosphorylation events to convert the stimuli into an appropriate cellular response. In this review, the variability of TCS domains and the most common signaling mechanisms are highlighted. This is followed by the description of the overall cellular topology, classification and mechanisms of MCP. Finally, the structural and functional properties of a new family of MCP found in Geobacter sulfurreducens are revisited. This bacterium has a diverse repertoire of chemosensory systems, which represents a striking example of a survival mechanism in challenging environments. Two G. sulfurreducens MCP—GSU0582 and GSU0935—are members of a new family of chemotaxis sensor proteins containing a periplasmic PAS-like sensor domain with a c-type heme. Interestingly, the cellular location of this domain opens new routes to the understanding of the redox potential sensing signaling transduction pathways.
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Protein Crystallization in a Microfluidic Contactor with Nafion®117 Membranes - Polino, M.; Rho, H. S.; Pina, M. P.; Mallada, R.; Carvalho, A. L.; Romão, M.J.; Coelhoso, Isabel; Gardeniers, J. G. E.; Crespo, J. G.; Portugal, Carla A. M. (Membranes, 2021)
Protein crystallization still remains mostly an empirical science, as the production of crystals with the required quality for X-ray analysis is dependent on the intensive screening of the best protein crystallization and crystal’s derivatization conditions. Herein, this demanding step was addressed by the development of a high-throughput and low-budget microfluidic platform consisting of an ion exchange membrane (117 Nafion® membrane) sandwiched between a channel layer (stripping phase compartment) and a wells layer (feed phase compartment) forming 75 independent micro-contactors. This microfluidic device allows for a simultaneous and independent screening of multiple protein crystallization and crystal derivatization conditions, using Hen Egg White Lysozyme (HEWL) as the model protein and Hg2+ as the derivatizing agent. This microdevice offers well-regulated crystallization and subsequent crystal derivatization processes based on the controlled transport of water and ions provided by the 117 Nafion® membrane. Diffusion coefficients of water and the derivatizing agent (Hg2+) were evaluated, showing the positive influence of the protein drop volume on the number of crystals and crystal size. This microfluidic system allowed for crystals with good structural stability and high X-ray diffraction quality and, thus, it is regarded as an efficient tool that may contribute to the enhancement of the proteins’ crystals structural resolution.
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Protein Engineering of Electron Transfer Components from Electroactive Geobacter Bacteria - Fernandes, Tomás M.; Leonor Morgado; Turner, David L.; Salgueiro, , Carlos A. (Antioxidants, 2021)
Electrogenic microorganisms possess unique redox biological features, being capable of transferring electrons to the cell exterior and converting highly toxic compounds into nonhazardous forms. These microorganisms have led to the development of Microbial Electrochemical Technologies (METs), which include applications in the fields of bioremediation and bioenergy production. The optimization of these technologies involves efforts from several different disciplines, ranging from microbiology to materials science. Geobacter bacteria have served as a model for understanding the mechanisms underlying the phenomenon of extracellular electron transfer, which is highly dependent on a multitude of multiheme cytochromes (MCs). MCs are, therefore, logical targets for rational protein engineering to improve the extracellular electron transfer rates of these bacteria. However, the presence of several heme groups complicates the detailed redox characterization of MCs. In this Review, the main characteristics of electroactive Geobacter bacteria, their potential to develop microbial electrochemical technologies and the main features of MCs are initially highlighted. This is followed by a detailed description of the current methodologies that assist the characterization of the functional redox networks in MCs. Finally, it is discussed how this information can be explored to design optimal Geobacter-mutated strains with improved capabilities in METs.
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Structural and functional insights of GSU0105, a unique multiheme cytochrome from G. sulfurreducens - Fernandes, Tomás M.; Folgosa, Filipe; Teixeira, Miguel; Salgueiro, , Carlos A.; Leonor Morgado (Biophysical Journal, 2021)
Geobacter sulfurreducens possesses over 100 cytochromes that assure an effective electron transfer to the cell exterior. The most abundant group of cytochromes in this microorganism is the PpcA family, composed of five periplasmic triheme cytochromes with high structural homology and identical heme coordination (His-His). GSU0105 is a periplasmic triheme cytochrome synthetized by G. sulfurreducens in Fe(III)-reducing conditions but is not present in cultures grown on fumarate. This cytochrome has a low sequence identity with the PpcA family cytochromes and a different heme coordination, based on the analysis of its amino acid sequence. In this work, amino acid sequence analysis, site-directed mutagenesis, and complementary biophysical techniques, including ultraviolet-visible, circular dichroism, electron paramagnetic resonance, and nuclear magnetic resonance spectroscopies, were used to characterize GSU0105. The cytochrome has a low percentage of secondary structural elements, with features of α-helices and β-sheets. Nuclear magnetic resonance shows that the protein contains three low-spin hemes (Fe(II), S = 0) in the reduced state. Electron paramagnetic resonance shows that, in the oxidized state, one of the hemes becomes high-spin (Fe(III), S = 5/2), whereas the two others remain low-spin (Fe(III), S = 1/2). The data obtained also indicate that the heme groups have distinct axial coordination. The apparent midpoint reduction potential of GSU0105 (−154 mV) is pH independent in the physiological range. However, the pH modulates the reduction potential of the heme that undergoes the low- to high-spin interconversion. The reduction potential values of cytochrome GSU0105 are more distinct compared to those of the PpcA family members, providing the protein with a larger functional working redox potential range. Overall, the results obtained, together with an amino acid sequence analysis of different multiheme cytochrome families, indicate that GSU0105 is a member of a new group of triheme cytochromes.
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A unique aromatic residue modulates the redox range of a periplasmic multiheme cytochrome from Geobacter metallireducens - Portela, Pilar C.; Silva, , Marta A.; Liliana R. Teixeira; Salgueiro, , Carlos A. (Journal of Biological Chemistry, 2021)
Geobacter bacteria are able to transfer electrons to the exterior of the cell and reduce extracellular electron acceptors including toxic/radioactive metals and electrode surfaces, with potential applications in bioremediation or electricity harvesting. The triheme c-type cytochrome PpcA from Geobacter metallireducens plays a crucial role in bridging the electron transfer from the inner to the outer membrane, ensuring an effective extracellular electron transfer. This cytochrome shares 80% identity with PpcA from Geobacter sulfurreducens, but their redox properties are markedly different, thus determining the distinctive working redox potential ranges in the two bacteria. PpcA from G. metallireducens possesses two extra aromatic amino acids (Phe-6 and Trp-45) in its hydrophobic heme core, whereas PpcA from G. sulfurreducens has a leucine and a methionine in the equivalent positions. Given the different nature of these residues in the two cytochromes, we have hypothesized that the extra aromatic amino acids could be partially responsible for the observed functional differences. In this work, we have replaced Phe-6 and Trp-45 residues by their nonaromatic counterparts in PpcA from G. sulfurreducens. Using redox titrations followed by UV–visible and NMR spectroscopy we observed that residue Trp-45 shifted the redox potential range 33% toward that of PpcA from G. sulfurreducens, whereas Phe-6 produced a negligible effect. For the first time, it is shown that the inclusion of an aromatic residue at the heme core can modulate the working redox range in abundant periplasmic proteins, paving the way to engineer bacterial strains for optimal microbial bioelectrochemical applications.
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Keywords: bioenergetics; Cytochrome; Electron transfer; NMR; Site-directed mutagenesis

Revisiting Ionic Liquid Structure-Property Relationship: A Critical Analysis - Silva, Wagner; Zanatta, Marcileia; Ferreira, Ana Sofia; Corvo, Marta C.; Cabrita, Eurico J. (International Journal of Molecular Sciences, 2020)
{In the last few years, ionic liquids (ILs) have been the focus of extensive studies concerning the relationship between structure and properties and how this impacts their application. Despite a large number of studies, several topics remain controversial or not fully answered, such as: the existence of ion pairs, the concept of free volume and the effect of water and its implications in the modulation of ILs physicochemical properties. In this paper, we present a critical review of state-of-the-art literature regarding structure-property relationship of ILs, we re-examine analytical theories on the structure-property correlations and present new perspectives based on the existing data. The interrelation between transport properties (viscosity, diffusion, conductivity) of IL structure and free volume are analysed and discussed at a molecular level. In addition, we demonstrate how the analysis of microscopic features (particularly using NMR-derived data) can be used to explain and predict macroscopic properties, reaching new perspectives on the properties and application of ILs.}
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Large-pore silica spheres as support for samarium-coordinated undecamolybdophosphate: Oxidative desulfurization of diesels - Mirante, Fatima; Alves, Ana C.; Juliao, Diana; Almeida, Pedro L; Gago, Sandra; Valenca, Rita; Ribeiro, Jorge C.; Baltazar de Castro; Granadeiro, Carlos M.; Balula, Salete S. (Fuel, 2020)
A novel composite has been prepared through the immobilization of the Keggin sandwich-type {[}Sm (PMo11O39)(2)](11-) anion (SmPOM) on large-pore silica spheres previously functionalized with trimethylammonium groups (TMA). The resulting SmPOM@TMA-LPMS material has been evaluated as heterogeneous catalyst in a biphasic desulfurization 1:1 diesel/extraction solvent system using H2O2 as oxidant. Preliminary experiments were conducted with different extraction solvents, acetonitrile and {[}BMIM]PF6 ionic liquid. The optimized extractive and catalytic oxidative desulfurization system (ECODS) with {[}BMIM]PF6 was able to reach complete sulfur removal from a model diesel containing 2100 ppm S in just 60 min (10 min of initial extraction + 50 min of catalytic step). The reutilization of catalyst and extraction phase has been successfully performed without loss of desulfurization efficiency in consecutive cycles, turning the process more sustainable and cog-effective. The remarkable results with simulated diesel have motivated the application of the catalyst in the desulfurization of untreated real diesel and 74% of efficiency was achieved after only 2 h for three consecutive cycles.
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Cleaning fungal stains on paper with hydrogels: The effect of pH control - Barrulas, V, Raquel; Nunes, Andre D.; Sequeira, Silvia O.; Casimiro, Maria Helena; Corvo, Marta C. (International Biodeterioration & Biodegradation, 2020)
{Fungal stains affect documents and artworks on paper all over the world, diminishing their chemical stability and compromising their readability. The present paper studies the suitability of agarose and gellan gum hydrogels to remove fungal stains from paper, using paper impregnated with alizarin as a model system to simulate the most common colorant molecules produced by fungi - polyketide quinones. The effect of pH variation on the efficacy of the gels was evaluated by UV spectrometry. The results show that the cleaning efficacy of the gels greatly depends on the gel matrix, the colorant molecules, and the pH balance of the process.}
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Polyoxometalate@Periodic mesoporous organosilicas as active materials for oxidative desulfurization of diesels - Ribeiro, Susana O.; Almeida, Pedro L; Pires, Joao; Baltazar de Castro; Balula, Salete S. (Microporous and Mesoporous Materials, 2020)
Novel material catalysts based in the active zinc-substituted polyoxotungstate ({[}PW11Zn(H2O)(39)](5-), abbreviated as PW11Zn) were efficiently used in the oxidative desulfurization of real and model diesels. These active catalytic center was strategically immobilized in a less hydrophilic periodic mesoporous organosilicas (PMOs), containing ethane-bridge (PMOE) and benzene-bridge (PMOB) walls, functionalized with (3-aminopropyl)triethoxysilane (aptes). The efficiency of the novel catalytic composites (PW11Zn@aptesPMOE and PM11Zn@aptesPMOB) was studied under oxidative desulfurization system (CODS) without the presence of an extraction solvent and also using a biphasic (diesel/extraction solvent) oxidative desulfurization system (ECODS). Both composites presented higher desulfurization efficiency under the solvent-free system, reaching ultra-low levels of sulfur compounds after only 1 h and using low ratio of H2O2/S = 4. The catalysts could be recycled without loss of activity for ten consecutive cycles. However, after the first desulfurization cycle complete desulfurization was achieved within only 30 min using PW11Zn@aptesPMOE composite. Also, the structure of PW it Zn@aptesPMOE demonstrated to be more stable than PW11Zn@aptesPMOB, probably due to the occurrence of some PW11Zn leaching from the PMOB surface, probably caused by the lower interaction of PW11Zn with the benzene-bridge PMOB wall. The most robust catalyst PW11Zn@aptesPMOE was used to desulfurize a real diesel achieving 75.9% of desulfurization after 2 h. The catalyst was further recycled with success to treat real diesel.
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The intracellular number of magnetic nanoparticles modulates the apoptotic death pathway after magnetic hyperthermia treatment - Beola, Lilianne; Laura Asín; Catarina Roma-Rodrigues; Yilian Fernandez-Afonso; Fratila, Raluca M.; Serantes, David; Ruta, Sergiu; Chantrell, Roy W.; Fernandes, Alexandra R; Baptista, Pedro V; {de la Fuente}, Jesus M.; Valeria Grazu; et al. (ACS Applied Materials & Interfaces, 2020)
Magnetic hyperthermia is a cancer treatment based on the exposure of magnetic nanoparticles to an alternating magnetic field in order to generate local heat. In this work, 3D cell culture models were prepared to observe the effect that a different number of internalized particles had on the mechanisms of cell death triggered upon the magnetic hyperthermia treatment. Macrophages were selected by their high capacity to uptake nanoparticles. Intracellular nanoparticle concentrations up to 7.5 pg Fe/cell were measured both by elemental analysis and magnetic characterization techniques. Cell viability after the magnetic hyperthermia treatment was decreased to <25% for intracellular iron contents above 1 pg per cell. Theoretical calculations of the intracellular thermal effects that occurred during the alternating magnetic field application indicated a very low increase in the global cell temperature. Different apoptotic routes were triggered depending on the number of internalized particles. At low intracellular magnetic nanoparticle amounts (below 1 pg Fe/cell), the intrinsic route was the main mechanism to induce apoptosis, as observed by the high Bax/Bcl-2 mRNA ratio and low caspase-8 activity. In contrast, at higher concentrations of internalized magnetic nanoparticles (1−7.5 pg Fe/cell), the extrinsic route was observed through the increased activity of caspase-8. Nevertheless, both mechanisms may coexist at intermediate iron concentrations. Knowledge on the different mechanisms of cell death triggered after the magnetic hyperthermia treatment is fundamental to understand the biological events activated by this procedure and their role in its effectiveness.
PCIN-2017-060 FCT/MCTES M-ERA NET/2/0008/2016 PGC2018-096016-B-I00 BIO2017-84246-C2-1-R UIDB/04378/2020 ED431E2018/08 EP/K031589/1 CAS18/00233 RYC-2014-15512 RYC-2015-17640 Sem PDF conforme despacho.
Keywords: 3D cell culture; Apoptosis; cell death pathways; iron oxides; Macrophages; Magnetic hyperthermia; nanoparticle uptake

Hyperthermia induced by gold nanoparticles and visible light photothermy combined with chemotherapy to tackle doxorubicin sensitive and resistant colorectal tumor 3D spheroids - Catarina Roma-Rodrigues; Pombo, Inês; Fernandes, Alexandra R; Baptista, Pedro V (International Journal of Molecular Sciences, 2020)
Current cancer therapies are frequently ineffective and associated with severe side effects and with acquired cancer drug resistance. The development of effective therapies has been hampered by poor correlations between pre-clinical and clinical outcomes. Cancer cell-derived spheroids are three-dimensional (3D) structures that mimic layers of tumors in terms of oxygen and nutrient and drug resistance gradients. Gold nanoparticles (AuNP) are promising therapeutic agents which permit diminishing the emergence of secondary effects and increase therapeutic efficacy. In this work, 3D spheroids of Doxorubicin (Dox)-sensitive and -resistant colorectal carcinoma cell lines (HCT116 and HCT116-DoxR, respectively) were used to infer the potential of the combination of chemotherapy and Au-nanoparticle photothermy in the visible (green laser of 532 nm) to tackle drug resistance in cancer cells. Cell viability analysis of 3D tumor spheroids suggested that AuNPs induce cell death in the deeper layers of spheroids, further potentiated by laser irradiation. The penetration of Dox and earlier spheroid disaggregation is potentiated in combinatorial therapy with Dox, AuNP functionalized with polyethylene glycol (AuNP@PEG) and irradiation. The time point of Dox administration and irradiation showed to be important for spheroids destabilization. In HCT116-sensitive spheroids, pre-irradiation induced earlier disintegration of the 3D structure, while in HCT116 Dox-resistant spheroids, the loss of spheroid stability occurred almost instantly in post-irradiated spheroids, even with lower Dox concentrations. These results point towards the application of new strategies for cancer therapeutics, reducing side effects and resistance acquisition.
UIDB/04378/2020 SFRH/BPD/124612/2016
Keywords: 3D spheroids; colorectal cancer; doxorubicin resistance; Gold nanoparticles; photothermy

Tackling Multidrug Resistance in Streptococci: From Novel Biotherapeutic Strategies to Nanomedicines - Alves-Barroco, Cinthia; Lorenzo Rivas-García; Fernandes, Alexandra R; Baptista, Pedro Viana (Frontiers in Microbiology, 2020)
The pyogenic streptococci group includes pathogenic species for humans and other animals and has been associated with enduring morbidity and high mortality. The main reason for the treatment failure of streptococcal infections is the increased resistance to antibiotics. In recent years, infectious diseases caused by pyogenic streptococci resistant to multiple antibiotics have been raising with a significant impact to public health and veterinary industry. The rise of antibiotic-resistant streptococci has been associated to diverse mechanisms, such as efflux pumps and modifications of the antimicrobial target. Among streptococci, antibiotic resistance emerges from previously sensitive populations as result of horizontal gene transfer or chromosomal point mutations due to excessive use of antimicrobials. Streptococci strains are also recognized as biofilm producers. The increased resistance of biofilms to antibiotics among streptococci promote persistent infection, which comprise circa 80% of microbial infections in humans. Therefore, to overcome drug resistance, new strategies, including new antibacterial and antibiofilm agents, have been studied. Interestingly, the use of systems based on nanoparticles have been applied to tackle infection and reduce the emergence of drug resistance. Herein, we present a synopsis of mechanisms associated to drug resistance in (pyogenic) streptococci and discuss some innovative strategies as alternative to conventional antibiotics, such as bacteriocins, bacteriophage, and phage lysins, and metal nanoparticles. We shall provide focused discussion on the advantages and limitations of agents considering application, efficacy and safety in the context of impact to the host and evolution of bacterial resistance.
UID/Multi/04378/2020 SFRH/BD/118350/2016 Inn-Indigo 0002/2015 RA Detect
Keywords: antimicrobial resistance; bacteriocins; bacteriophage; biofilms; Nanomedicine; nanoparticles; pyogenic streptococci

CO2 removal from anaesthesia circuits using gas-ionic liquid membrane contactors - Martins, C. F.; Neves, L. A.; Chagas, R.; Ferreira, LM; C. A. M. Afonso; Crespo, J. G.; Coelhoso, I. M. (SEPARATION AND PURIFICATION TECHNOLOGY, 2020)
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Correction to: Nano-in-Micro Sildenafil Dry Powder Formulations for the Treatment of Pulmonary Arterial Hypertension Disorders: The Synergic Effect of POxylated Polyurea Dendrimers, PLGA, and Cholesterol (Part. Part. Syst. Charact, (2020), 37, (1900447), - Restani, Rita B.; Tavares, Márcia T.; Pires, Rita F.; Baptista, Pedro V; Fernandes, Alexandra R; Casimiro, Teresa; Bonifácio, Vasco D. B.; Aguiar-Ricardo, Ana (Particle and Particle Systems Characterization, 2020)
Part. Part. Syst. Charact. 2020, 37, 1900447 In the originally published manuscript, the author Márcia T. Tavares was omitted. The author is hereby added in the author byline and is associated with the first affiliation.
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Water safety screening via multiplex LAMP-Au-nanoprobe integrated approach - Oliveira, Beatriz B.; Bruno Veigas; Carlos, Fábio Ferreira; Sánchez-Melsió, Alexandre; Balcázar, José Luís; Borrego, Carles M.; Baptista, Pedro Viana (Science of the Total Environment, 2020)
Contaminated water resources remain a major global concern regarding public health. The majority of water safety protocols include indicators of microbial contamination to evaluate the potential risk to public health and are key elements of quality guidelines. Among these, markers for total coliforms and fecal coliforms are strong indicators of co-contamination with other pathogens. Traditional methods, recurring to slow and cumbersome culture-based approaches, have been gradually replaced by molecular methods, capable of faster and more specific screening. These are usually PCR-based methods that may allow for multiple pathogen detection but require dedicated laboratory equipment, hindering the rapid on-site assessment. Here, we used a multiplex Loop-Mediated Isothermal Amplification (mLAMP) strategy for the amplification of two markers associated with the contamination by total and fecal coliforms (e.g. Escherichia coli) — lacZ and uidA genes, respectively — thus allowing for single tube multiplex detection. The mLAMP products were then subject to an Au-nanoprobe colorimetric detection assay for precise discrimination of targets. This approach was validated in 22 water samples that were also screened for the presence of lacZ and uidA using standard and quantitative PCR, with the capability for discriminating the contamination level, e.g. a semi-quantitative evaluation of water quality.
This work was financed by FEDER funds through the COMPETE 2020 Program and National Funds through FCT (Portuguese Foundation for Science and Technology) under the projects UlD/Multi/04378/2013 and UID/Multi/04378/2019 for Applied Molecular Biosciences Unit (UCIBIO), SFRH/BPD/124311/2016. JPI Proj No 281715 (TRACE) project. We also acknowledge Sigita Stanynaite for the restriction analysis of lacZ and uidA LAMP products. ICRA researchers acknowledge the support for scientific equipment given by the European Regional Development Fund (FEDER) under the Catalan FEDER Operative Program 2007-2013 and by MINECO according to DA3a of the Catalan Statute of Autonomy and to PGE-2010 as well as the funding from the CERCA program of the Catalan government.
Keywords: E. coli; loop-mediated isothermal amplification; Point-of-need screening; Water safety screening

Fast prototyping microfluidics: Integrating droplet digital lamp for absolute quantification of cancer biomarkers - Beatriz Oliveira; Bruno Veigas; Fernandes, Alexandra R; Hugo Águas; Rodrigo Martins; Elvira Fortunato; Baptista, Pedro Viana (Sensors, 2020)
Microfluidic (MF) advancements have been leveraged toward the development of state-of-the-art platforms for molecular diagnostics, where isothermal amplification schemes allow for further simplification of DNA detection and quantification protocols. The MF integration with loop-mediated isothermal amplification (LAMP) is today the focus of a new generation of chip-based devices for molecular detection, aiming at fast and automated nucleic acid analysis. Here, we combined MF with droplet digital LAMP (ddLAMP) on an all-in-one device that allows for droplet generation, target amplification, and absolute quantification. This multilayer 3D chip was developed in less than 30 minutes by using a low-cost and extremely adaptable production process that exploits direct laser writing technology in “Shrinky-dinks” polystyrene sheets. ddLAMP and target quantification were performed directly on-chip, showing a high correlation between target concentration and positive droplet score. We validated this integrated chip via the amplification of targets ranging from five to 500,000 copies/reaction. Furthermore, on-chip amplification was performed in a 10 µL volume, attaining a limit of detection of five copies/µL under 60 min. This technology was applied to quantify a cancer biomarker, c-MYC, but it can be further extended to any other disease biomarker.
UID/CTM/50025/2019 UID/Multi/04378/2019 Inn-INDIGO/0002/2015 PTDC/BTM-SAL/31201/2017 SFRH/BPD/124311/2016
Keywords: digital amplification; integrated microfluidics device; lab-on-a-chip; loop-mediated isothermal amplification; oncogenes

Gene therapy in cancer treatment: Why go nano? - Catarina Roma-Rodrigues; Lorenzo Rivas-García; Baptista, Pedro V; Fernandes, Alexandra R (Pharmaceutics, 2020)
The proposal of gene therapy to tackle cancer development has been instrumental for the development of novel approaches and strategies to fight this disease, but the efficacy of the proposed strategies has still fallen short of delivering the full potential of gene therapy in the clinic. Despite the plethora of gene modulation approaches, e.g., gene silencing, antisense therapy, RNA interference, gene and genome editing, finding a way to efficiently deliver these effectors to the desired cell and tissue has been a challenge. Nanomedicine has put forward several innovative platforms to overcome this obstacle. Most of these platforms rely on the application of nanoscale structures, with particular focus on nanoparticles. Herein, we review the current trends on the use of nanoparticles designed for cancer gene therapy, including inorganic, organic, or biological (e.g., exosomes) variants, in clinical development and their progress towards clinical applications.
This work was supported by the Applied Molecular Biosciences Unit -UCIBIO which is financed by national funds from FCT (UIDB/04378/2020); CRR (SFRH/BPD/124612/2016) and LRG (Inn-Indigo 00002/2015 RA Detect).
Keywords: gene delivery; Gene therapy; Nanomedicine; nanoparticles; Tumor microenvironment

Synthesis and characterization of dicarboxymethyl cellulose - Chagas, Ricardo; Gericke, Martin; Ferreira, Ricardo B.; Heinze, Thomas; Ferreira, Luisa M. (CELLULOSE, 2020)
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Nano-in-Micro Sildenafil Dry Powder Formulations for the Treatment of Pulmonary Arterial Hypertension Disorders: The Synergic Effect of POxylated Polyurea Dendrimers, PLGA, and Cholesterol - Restani, Rita B.; Pires, Rita F.; Baptista, Pedro V; Fernandes, Alexandra R; Casimiro, Teresa; Bonifácio, Vasco D. B.; Aguiar-Ricardo, Ana (Particle and Particle Systems Characterization, 2020)
POXylated polyurea dendrimer nanoparticles (PUREG4OOx48) are loaded with sildenafil (SDF) by a supercritical carbon dioxide–assisted (scCO2) impregnation. Further supercritical CO2-assisted spray drying (SASD) leads to hybrid nano-in-micro dry powder formulations that are investigated aiming at efficient pulmonary delivery of SDF in pulmonary arterial hypertension treatment. This is the first report of the production of poly(D,L-lactide-co-glycolide)-cholesterol (PLGA-Chol) microparticles processed by SASD. The optimized formulation of nano-in-microparticles is composed of PLGA, Chol, and PUREG4OOx48, loaded with SDF solutions in a 77:23 ratio (PLGA-Chol:dendrimer, w/w). The dry powders are fully characterized and found to be highly biodegradable and biocompatible, and the SDF release profile evaluates under different pH values. The median mass average diameter (MMAD) of the nano-in-micro systems varies between 2.57 and 5 µm and the fine particle fraction (FPF) between 36% and 29% for PUREG4OMeOx48[PLGA-Chol] and PUREG4OEtOx48[PLGA-Chol], respectively. The data validate the potential use of these new formulations in inhalation therapy. In vitro studies are also carried out in order to evaluate the effect of the free drug in cell viability and formulations cytotoxicity.
info:eu-repo/grantAgreement/FCT/3599-PPCDT/99452/PT# info:eu-repo/grantAgreement/FCT/SFRH/SFRH%2FBD%2F66858%2F2009/PT# UID/QUI/50006/2020 POCI-01-0145-FEDER-007265 UID/Multi/04378/2020 SFRH/BD/109006/2015 Sem PDF conforme despacho
Keywords: cholesterol; nano-in-microparticles; PLGA; Pulmonary drug delivery; sildenafil citrate; supercritical-assisted spray drying

Synthesis of new hetero-arylidene-9(10H)-anthrone derivatives and their biological evaluation - Catarina Roma-Rodrigues; Malta, Gabriela; Peixoto, Daniela; Ferreira, Luísa M.; Baptista, Pedro V; Fernandes, Alexandra R; Branco, Paula S. (Bioorganic Chemistry, 2020)
New hetero-arylidene-9(10H)-anthrone derivatives (1) were synthesized from reaction of 1,2-dimethyl-3-alkyl imidazolium salts (2) and 9-anthracenecarboxaldehyde. Ion exchange of the anion with dioctyl sulfosuccinate and lithium bis(trifluoromethanesulfonyl)imide led to the preparation of other derivatives. The antiproliferative effect of the compounds was evaluated in human ovarian (A2780) and colorectal (HCT116) carcinoma cell lines and in normal primary human fibroblasts. Compound 1 presented an antiproliferative effect related to the imidazolium pattern of substitution with compounds having a decyl group at the R-position (1c and 3c) showing the highest cytotoxic activities in all cell lines independently of the counter ion. Compounds 1b and 1c internalize A2780 cancer cells via a passive or an active transport, respectively, inducing A2780 cell death via an extrinsic apoptosis (1b) or intrinsic apoptosis and oncosis (1c). The localization of both compounds in the cytoplasm coupled to the absence of reactive oxygen species (ROS) induction suggest that the mechanisms of toxicity might be different than those of other anthracyclines currently used in chemotherapy.
info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F50006%2F2020/PT# info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F04378%2F2020/PT# info:eu-repo/grantAgreement/FCT/3599-PPCDT/RECI%2FBBB-BQB%2F0230%2F2012/PT# info:eu-repo/grantAgreement/FCT/3599-PPCDT/RECI%2FBBB-BEP%2F0124%2F2012/PT#
Keywords: Arylidene-anthrone derivatives; Cell apoptosis; Imidazolium chemistry; Ovarian cancer; Proliferation inhibition

Synthesis of new hetero-arylidene-9(10H)-anthrone derivatives and their biological evaluation - Catarina Roma-Rodrigues; Malta, Gabriela; Peixoto, Daniela; Ferreira, Luisa M.; Pedro Baptista; Alexandra R. Fernandes; Branco, Paula S. (BIOORGANIC CHEMISTRY, 2020)
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Light Irradiation of Gold Nanoparticles Toward Advanced Cancer Therapeutics - Amendoeira, Ana; García, Lorenzo Rivas; Fernandes, Alexandra R; Baptista, Pedro V (Advanced Therapeutics, 2020)
Cancer is one of the leading causes of death in the world. To challenge this epidemic, there are growing demands for the development of new advanced and targeted therapeutics capable of effectively tackling cancer cells with improved selectivity. Nanomedicine has put forward several innovative therapeutics toward improving therapeutic efficacy while decreasing the deleterious side effects of current chemotherapy. Multifunctional gold nanoparticles (AuNPs) have been at the core of a plethora of advanced therapeutic strategies that provide selective targeting with their unique optical properties, capable to interact with the light of specific wavelength to deliver therapy with tremendous spatiotemporal precision. AuNPs have been exploited as photodynamic and photothermal therapeutic agents alone or in combination with other cancer treatment modalities with other cancer applications. Due to their exceptional physicochemical properties, they have been proven efficacious allies for photodynamic therapy and for photothermal therapy regimens. Herein, the rapidly progressing literature related to the use of these promising strategies against cancer is discussed, highlighting their possible future clinical translation.
Funding Information: info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UID%2FMulti%2F04378%2F2019/PT# Publisher Copyright: © 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Keywords: Cancer; Gold nanoparticles; Nanomedicine; photodynamic therapy; photothermal therapy

Nanotheranostics in Gene Therapy - Oliveira, Beatriz B.; Fernandes, Alexandra R; Baptista, Pedro V (Advances in Cancer Nanotheranostics for Experimental and Personalized Medicine, 2020)
The continuous advances in molecular genetics have prompt for a wealth of tools capable to modulate genome and the corresponding gene expression. These innovative technologies have broadened the range of possibilities for gene therapy, either to decrease expression of malignant genes and mutations or edition of genomes for correction of errors. These strategies rely on the delivery of therapeutic nucleic acids to cells and tissues that must overcome several biological barriers. Indeed, a key element for the success of any gene therapy formulation is the carrier agent capable to deliver the therapeutic nucleic acid moieties to a specific target and promote efficient cellular uptake, while preventing deleterious off-target effects and degradation by endogenous nucleases. The initial vectorization strategies proved to be rather immunogenic, limited in the amount of genetic material that can be packed and raised severe toxicity concerns. Nowadays, a new generation of nanotechnology-based gene delivery systems are making an impact on the way we use therapeutic nucleic acids. These nanovectorization platforms have been developed so as to show low immunogenicity, low toxicity, ease of assembly and scale-up with higher loading capacity. Some of these nanoscale systems have also allowed for controlled release system and for the simultaneous capability of monitorization of effect - nanotheranostics. Herein, we provide a review on the variety of gene delivery vectors and platforms at the nanoscale.
Publisher Copyright: © 2020, Bentham Books imprint.
Keywords: Cancer; Gene therapy; Nanomedicine; nanotheranostics; RNAi

A Novel Cellulose-Based Polymer for Efficient Removal of Methylene Blue - Gago, Diana; Chagas, Ricardo; Ferreira, Luisa M.; Velizarov, Svetlozar; Coelhoso, Isabel (MEMBRANES, 2020)
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Porphyrin Pigments in Polychaeta: Explorations on the Evolution of Haem Metabolism in Marine Eumetazoans - Martins, C.; Rodrigo, A. P.; Madeira, C; D'Ambrosio, M; Goncalves, C; Parola, A. J.; Grosso, A. R.; Baptista, P. V.; Fernandes, A. R.; Costa, P. M. (2020)
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Synthetic Approaches to a Challenging and Unusual Structure-An Amino-Pyrrolidine Guanine Core - Rippel, Rafael; Pinheiro, Luis; Lopes, Monica; Lourenco, Ana; Ferreira, Luisa M.; Branco, Paula S. (MOLECULES, 2020)
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Antiproliferative Activities of Diimine-Based Mixed Ligand Copper(II) Complexes - Kordestani, Nazanin; Rudbari, Hadi Amiri; Fernandes, Alexandra R; Raposo, Luís R.; Baptista, Pedro V; Ferreira, Daniela A.; Bruno, Giuseppe; Bella, Giovanni; Scopelliti, Rosario; Braun, Jason D.; Herbert, David E.; Olivier Blacque (ACS Combinatorial Science, 2020)
A series of Cu(diimine)(X-sal)(NO3) complexes, where the diimine is either 2,2′-bipyridine (bpy) or 1,10-phenanthroline (phen) and X-sal is a monoanionic halogenated salicylaldehyde (X = Cl, Br, I, or H), have been synthesized and characterized by elemental analysis and X-ray crystallography. Penta-coordinate geometries copper(II) were observed for all cases. The influence of the diimine coligands and different halogen atoms on the antiproliferative activities toward human cancer cell lines have been investigated. All Cu(II) complexes were able to induce a loss of A2780 ovarian carcinoma cell viability, with phen derivatives more active than bpy derivatives. In contrast, no in vitro antiproliferative effects were observed against the HCT116 colorectal cancer cell line. These cytotoxicity differences were not due to a different intracellular concentration of the complexes determined by inductively coupled plasma atomic emission spectroscopy. A small effect of different halogen substituents on the phenolic ring was observed, with X = Cl being the most highly active toward A2780 cells among the phen derivatives, while X = Br presented the lowest IC50 in A2780 cells for bpy analogs. Importantly, no reduction in normal primary fibroblasts cell viability was observed in the presence of bpy derivatives (IC50 > 40 μM). Mechanistically, complex 1 seems to induce a stronger apoptotic response with a higher increase in mitochondrial membrane depolarization and an increased level of intracellular reactive oxygen species (ROS) compared to complex 3. Together, these data and the low IC50 compared to cisplatin in A2780 ovarian carcinoma cell line demonstrate the potential of these bpy derivatives for further in vivo studies.
FCT/MCTES (UID/Multi/04378/2019 project PTDC/CVT/EPI/6685/2014 M-ERA/NET2/0008/2016 Sem PDF conforme despacho.
Keywords: Apoptosis; Cancer; copper(II); Crystal structure; cytotoxicity; halogenated salicylaldehyde; mixed ligands; ovarian carcinoma cells

Identification of pheromone candidates for the eucalyptus weevil, Gonipterus platensis (Coleoptera, Curculionidae) - Branco, Sofia; Mateus, Eduardo Pires; Richter Gomes da Silva, Marco Diogo; Mendes, Davide; Araujo Pereira, Maria Manuela; Schutz, Stefan; Paiva, Maria Rosa (JOURNAL OF APPLIED ENTOMOLOGY, 2020)
The eucalyptus weevil, Gonipterus platensis (Coleoptera, Curculionidae), is a major pest of eucalyptus plantations worldwide. To date, no pheromones have been identified for this species, despite their valuable potential as tools in monitoring or control strategies. Here we report the detection and identification of pheromones candidates of G. platensis. The weevil's volatile compounds were collected by solid phase micro extraction (SPME) and monolithic material sorption extraction (MMSE). Using Gas Chromatography coupled to Mass Spectrometry (GC/MS) analysis, eleven insect specific compounds were detected and identified: verbenene, cis-verbenol, trans-verbenol, verbenone, 2-oxo-1,8-cineole, 9-hydroxy-1,8-cineole, 2-alpha-hydroxy-1,8-cineole, 3-oxo-1,8-cineole, 2-beta-hydroxy-1,8-cineole, 3-alpha-hydroxy-1,8-cineole and 7-hydroxy-1,8-cineole. Three of these compounds, verbenene, cis-verbenol and trans-verbenol, were shown to be male-specific. Antennal sensitivity towards ten compounds emitted by G. platensis was detected using Gas Chromatography-Mass Spectrometry/Electroantennographic Detection (GC-MS/EAD). Extracts from virgin males proved to be attractive to virgin females in olfactometer bioassays. Further behavioural bioassays showed that both virgin females and virgin males were attracted to the male-specific compound cis-verbenol and that virgin females were attracted to trans-verbenol. Verbenone was attractive to mated females. Regarding 2-alpha-hydroxy-1.8-cineole and 2-oxo-1,8-cineole, which are produced by both sexes, the alcohol was attractive to virgin males and both the alcohol and the ketone were repellant to mated females. This is, to our knowledge, the first identification of pheromones candidates in Gonipterus spp. and also the first evidence of cineole metabolites acting as semiochemicals.
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Improving the Anti-inflammatory Response via Gold Nanoparticle Vectorization of CO-Releasing Molecules - Fernandes, Alexandra R; Inês Mendonça-Martins; Santos, Marino F. A.; Raposo, Luís R.; Rita Mendes; Marques, Joana; Romão, Carlos C.; Romão, Maria João; Santos-Silva, Teresa; Baptista, Pedro V (ACS Biomaterials Science and Engineering, 2020)
CO-releasing molecules (CORMs) have been widely studied for their anti-inflammatory, antiapoptotic, and antiproliferative effects. CORM-3 is a water-soluble Ru-based metal carbonyl complex, which metallates serum proteins and readily releases CO in biological media. In this work, we evaluated the anti-inflammatory and wound-healing effects of gold nanoparticles-CORM-3 conjugates, AuNPs@PEG@BSA·Ru(CO)x, exploring its use as an efficient CO carrier. Our results suggest that the nanoformulation was capable of inducing a more pronounced cell effect, at the anti-inflammatory level and a faster tissue repair, probably derived from a rapid cell uptake of the nanoformulation that results in the increase of CO inside the cell.
UID/Multi/04378/2019 FCT/MEC PTDC/QEQMED/1902/2014 Sem PDF conforme despacho.
Keywords: anti-inflammatory effects; bovine serum albumin; CO-releasing molecules; CORM-3; Gold nanoparticles; tissue regeneration

Size-Dependent Biological Activities of Fluorescent Organosilane-Modified Zinc Oxide Nanoparticles - Mariana Buşilă; Aurel Tăbăcaru; Viorica Muşsat; Vasile, Bogdan Ştefan; Neaşu, Ionela Andreea; Pinheiro, Teresa; Catarina Roma-Rodrigues; Baptista, Pedro V; Fernandes, Alexandra R; Matos, António Pedro; Marques, Fernanda Marujo (Journal of biomedical nanotechnology, 2020)
Surface modification of zinc oxide nanoparticles (ZnO NPs) is a strategy to tune their biocompatibility. Herein we report on the synthesis of a series of fluorescent ZnO NPs modified with 2-10% (3-glycidyloxypropyl)trimethoxysilane (GPTMS) to investigate the fluorescence properties and to explore their applications in microbiology and biomedicine. The obtained ZnO NPs were characterized by X-ray diffraction (XRD), high resolution transmission electron microscopy (HRTEM) and Fourier transform infrared spectroscopy (FTIR). Size reduction occurred from ca. 13 nm in unmodified ZnO to 3-4 nm in silane-modified samples and fluorescence spectra showed size-dependent variation of the photoemission bands' intensity. The antibacterial and cytotoxic activities were investigated on Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacteria, and in ovarian (A2780) and prostate (PC3) cancer cells by tetrazolium/formazan-based methods. The antibacterial effect was higher for E. coli than S. aureus, while the cytotoxic activity was similar for both cancer cells and varied with the particle size. Cell death by apoptosis, and/or necrosis versus autophagy, were explored by flow cytometry using an Annexin V based-method and transmission electron microscopy (TEM). The main mechanism of ZnO NPs toxicity may involve the generation of reactive oxygen species (ROS) and the induction of apoptosis or autophagy. This work revealed the potential utility of GPTMS-modified ZnO NPs in the treatment of bacterial infection and cancer.
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Supramolecular protein polymers using mini-ferritin Dps as the building block - Pacheco, M. Raquel; Jacinto, Joao P.; Penas, Daniela; Calmeiro, Tomas; Almeida, Ana; Colaco, Miriam; Elvira Fortunato; Jones, Nykola C.; Hoffmann, V, Soren; Pereira, M Manuela A; Tavares, Pedro; Pereira, Alice S. (ORGANIC & BIOMOLECULAR CHEMISTRY, 2020)
A missense mutant of a Dps protein (DNA-binding protein from starved cells) from Marinobacter hydrocarbonoclasticus was used as a building block to develop a new supramolecular assembly complex which enhances the iron uptake, a physiological function of this mini-ferritin. The missense mutation was conducted in an exposed and flexible region of the N-terminal, wherein a threonine residue in position 10 was replaced by a cysteine residue (DpsT10C). This step enabled a click chemistry approach to the variant DpsT10C, where a thiol-ene coupling occurs. Two methods and two types of linker were used resulting in two different mini-ferritin supramolecular polymers, which have maintained secondary structure and native iron uptake physiological function. Electrophoretic assays and mass spectrometry were utilized to confirm that both functionalization and coupling reactions occured as predicted. The secondary structure has been investigated by circular dichroism and synchrotron radiation circular dichroism. Size and morphology were obtained by dynamic light scattering, size exclusion chromatography and atomic force microscopy, respectively. The iron uptake of the synthesized protein polymers was confirmed by UV-Vis spectroscopy loading assays.
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Gold nanoparticles for vectorization of nucleic acids for cancer therapeutics - Ferreira, Daniela; Fontinha, David; Martins, Catarina; Pires, David; Fernandes, Alexandra R; Baptista, Pedro V (Molecules, 2020)
Cancer remains a complex medical challenge and one of the leading causes of death worldwide. Nanomedicines have been proposed as innovative platforms to tackle these complex diseases, where the combination of several treatment strategies might enhance therapy success. Among these nanomedicines, nanoparticle mediated delivery of nucleic acids has been put forward as key instrument to modulate gene expression, be it targeted gene silencing, interference RNA mechanisms and/or gene edition. These novel delivery systems have strongly relied on nanoparticles and, in particular, gold nanoparticles (AuNPs) have paved the way for efficient delivery systems due to the possibility to fine-tune their size, shape and surface properties, coupled to the ease of functionalization with different biomolecules. Herein, we shall address the different molecular tools for modulation of expression of oncogenes and tumor suppressor genes and discuss the state-of-the-art of AuNP functionalization for nucleic acid delivery both in vitro and in vivo models. Furthermore, we shall highlight the clinical applications of these spherical AuNP based conjugates for gene delivery, current challenges, and future perspectives in nanomedicine.
info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F04378%2F2020/PT# info:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FNAN-MAT%2F31100%2F2017/PT# info:eu-repo/grantAgreement/FCT/3599-PPCDT/M-ERA-NET2%2F0008%2F2016/PT#
Keywords: gene editing; Gene silencing; Gold nanoparticles; Nanomedicine; therapeutic nucleic acid

Invasive Plants: Turning Enemies into Value - Maximo, Patricia; Ferreira, Luisa M.; Branco, Paula S.; Lourenco, Ana (MOLECULES, 2020)
In this review, a brief description of the invasive phenomena associated with plants and its consequences to the ecosystem is presented. Five worldwide invasive plants that are a threat to Portugal were selected as an example, and a brief description of each is presented. A full description of their secondary metabolites and biological activity is given, and a resume of the biological activity of extracts is also included. The chemical and pharmaceutical potential of invasive species sensu lato is thus acknowledged. With this paper, we hope to demonstrate that invasive species have potential positive attributes even though at the same time they might need to be controlled or eradicated. Positive attributes include chemical and pharmaceutical properties and developing these could help mitigate the costs of management and eradication.
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Colloidal lithography for transparent electronics and light trapping in thin film flexible solar cells - Mendes, Manuel J. (Encontro Ciência 2020, 2020)

All-cellulose composite membranes for oil microdroplet collection - Ana P.C. Almeida; Oliveira, João; Fernandes, Susete N.; Godinho, Maria H; Canejo, João P (2020)
Oil spills on ocean waters represent a major threat to marine ecosystems. A significant part of the spilled oil is dispersed in microdroplets that are not recovered by traditional oil-removing methods. In this work, we report on the manufacture of cellulose acetate (CA) electrospun non-woven membranes, stamped with different cellulose nanocrystal (CNC) patterns. We demonstrate the use of the membranes produced as selective oil microdroplets removal from water emulsions with an efficiency up to 80%. Screenprinting was used to imprint different CNC designs on the CA surface membranes. To promote the adhesion between the CNCs and the CNCs with the CA fibers the membrane was subjected to a thermal and chemical treatments. Oil droplets were collected under water in the oleophilic CNC pattern while the water could flow through the hydrophilic CA electrospun non-woven membrane.
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Antiproliferative Activities of Diimine-Based Mixed Ligand Copper(II) Complexes - Kordestani, N.; Rudbari, H. A.; Fernandes, A. R.; Raposo, L. R.; P.V. Baptista; Ferreira, D.; Bruno, G.; Bella, G.; Scopelliti, R.; Braun, J. D.; Herbert, D. E.; Blacque, O. (ACS Comb Sci, 2020)
A series of Cu(diimine)(X-sal)(NO3) complexes, where the diimine is either 2,2'-bipyridine (bpy) or 1,10-phenanthroline (phen) and X-sal is a monoanionic halogenated salicylaldehyde (X = Cl, Br, I, or H), have been synthesized and characterized by elemental analysis and X-ray crystallography. Penta-coordinate geometries copper(II) were observed for all cases. The influence of the diimine coligands and different halogen atoms on the antiproliferative activities toward human cancer cell lines have been investigated. All Cu(II) complexes were able to induce a loss of A2780 ovarian carcinoma cell viability, with phen derivatives more active than bpy derivatives. In contrast, no in vitro antiproliferative effects were observed against the HCT116 colorectal cancer cell line. These cytotoxicity differences were not due to a different intracellular concentration of the complexes determined by inductively coupled plasma atomic emission spectroscopy. A small effect of different halogen substituents on the phenolic ring was observed, with X = Cl being the most highly active toward A2780 cells among the phen derivatives, while X = Br presented the lowest IC50 in A2780 cells for bpy analogs. Importantly, no reduction in normal primary fibroblasts cell viability was observed in the presence of bpy derivatives (IC50 > 40 muM). Mechanistically, complex 1 seems to induce a stronger apoptotic response with a higher increase in mitochondrial membrane depolarization and an increased level of intracellular reactive oxygen species (ROS) compared to complex 3. Together, these data and the low IC50 compared to cisplatin in A2780 ovarian carcinoma cell line demonstrate the potential of these bpy derivatives for further in vivo studies.
Kordestani, NazaninRudbari, Hadi Amiri Fernandes, Alexandra R Raposo, Luis R Baptista, Pedro V Ferreira, Daniela Bruno, Giuseppe Bella, Giovanni Scopelliti, Rosario Braun, Jason D Herbert, David E Blacque, Olivier eng Research Support, Non-U.S. Gov't ACS Comb Sci. 2020 Feb 10;22(2):89-99. doi: 10.1021/acscombsci.9b00202. Epub 2020 Jan 23.
Keywords: *Apoptosis; *cancer; *copper(II); *crystal structure; *cytotoxicity; *halogenated salicylaldehyde; *mixed ligands; *ovarian carcinoma cells

Backbone, side chain and heme resonance assignment of the triheme cytochrome PpcA from Geobacter metallireducens in the oxidized state - Portela, Pilar C.; Joana M Dantas; Salgueiro, , Carlos A. (Biomol NMR Assign, 2020)
The bacterium Geobacter metallireducens is capable of transferring electrons to the cell exterior, a process designated extracellular electron transfer. This mechanism allows the microorganism to reduce extracellular acceptors such as Fe(III) (hydr)oxides and water toxic and/or radioactive contaminants including Cr(VI) and U(VI). It is also capable of oxidizing waste water aromatic organic compounds being an important microorganism for bioremediation of polluted waters. Extracellular electron transfer also allows electricity harvesting from microbial fuel cells, a promising sustainable form of energy production. However, extracellular electron transfer processes in this microorganism are still poorly characterized. The triheme c-type cytochrome PpcA from G. metallireducens is abundant in the periplasm and is crucial for electron transfer between the cytoplasm and the cell’s exterior. In this work, we report near complete assignment of backbone, side chain and heme resonances for PpcA in the oxidized state that will permit its structure determination and identification of interactions with physiological redox partners.
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Catalytic Mechanism of Human Aldehyde Oxidase - Ferreira, Pedro; Cerqueira, Nuno M. F. Sousa A.; Fernandes, Pedro Alexandrino; Romão, Maria João; Ramos, Maria João (ACS CatalysisACS Catalysis, 2020)
The mechanism of oxidation of N-heterocycle phthalazine to phthalazin-1(2H)-one and its associated free energy profile, catalyzed by human aldehyde oxidase (hAOX1), was studied in atomistic detail using QM/MM methodologies. The studied reaction was found to involve three sequential steps: (i) protonation of the substrate’s N2 atom by Lys893, (ii) nucleophilic attack of the hydroxyl group of the molybdenum cofactor (Moco) to the substrate, and (iii) hydride transfer from the substrate to the sulfur atom of the Moco. The free energy profile that was calculated revealed that the rate-limiting step corresponds to hydride transfer. It was also found that Lys893 plays a relevant role in the reaction, being important not only for the anchorage of the substrate close to the Moco, but also in the catalytic reaction. The variations of the oxidation state of the molybdenum ion throughout the catalytic cycle were examined too. We found out that during the displacement of the products away from the Moco, the transfer of electrons from the catalytic site to the FAD site was proton-coupled. As a consequence, the most favorable and fastest pathway for the enzyme to complete its catalytic cycle was that with MoV and a deprotonated SH ligand of the Moco with the FAD molecule converted to its semiquinone form, FADH•.The mechanism of oxidation of N-heterocycle phthalazine to phthalazin-1(2H)-one and its associated free energy profile, catalyzed by human aldehyde oxidase (hAOX1), was studied in atomistic detail using QM/MM methodologies. The studied reaction was found to involve three sequential steps: (i) protonation of the substrate’s N2 atom by Lys893, (ii) nucleophilic attack of the hydroxyl group of the molybdenum cofactor (Moco) to the substrate, and (iii) hydride transfer from the substrate to the sulfur atom of the Moco. The free energy profile that was calculated revealed that the rate-limiting step corresponds to hydride transfer. It was also found that Lys893 plays a relevant role in the reaction, being important not only for the anchorage of the substrate close to the Moco, but also in the catalytic reaction. The variations of the oxidation state of the molybdenum ion throughout the catalytic cycle were examined too. We found out that during the displacement of the products away from the Moco, the transfer of electrons from the catalytic site to the FAD site was proton-coupled. As a consequence, the most favorable and fastest pathway for the enzyme to complete its catalytic cycle was that with MoV and a deprotonated SH ligand of the Moco with the FAD molecule converted to its semiquinone form, FADH•.
doi: 10.1021/acscatal.0c02627

Evolution, expression, and substrate specificities of aldehyde oxidase enzymes in eukaryotes - Terao, Mineko; Garattini, Enrico; Romão, Maria João; Leimkühler, Silke (Journal of Biological ChemistryJournal of Biological Chemistry, 2020)
Aldehyde oxidases (AOXs) are a small group of enzymes belonging to the larger family of molybdo-flavoenzymes, along with the well-characterized xanthine oxidoreductase. The two major types of reactions that are catalyzed by AOXs are the hydroxylation of heterocycles and the oxidation of aldehydes to their corresponding carboxylic acids. Different animal species have different complements of AOX genes. The two extremes are represented in humans and rodents; whereas the human genome contains a single active gene (AOX1), those of rodents, such as mice, are endowed with four genes (Aox1-4), clustering on the same chromosome, each encoding a functionally distinct AOX enzyme. It still remains enigmatic why some species have numerous AOX enzymes, whereas others harbor only one functional enzyme. At present, little is known about the physiological relevance of AOX enzymes in humans and their additional forms in other mammals. These enzymes are expressed in the liver and play an important role in the metabolisms of drugs and other xenobiotics. In this review, we discuss the expression, tissue-specific roles, and substrate specificities of the different mammalian AOX enzymes and highlight insights into their physiological roles.Aldehyde oxidases (AOXs) are a small group of enzymes belonging to the larger family of molybdo-flavoenzymes, along with the well-characterized xanthine oxidoreductase. The two major types of reactions that are catalyzed by AOXs are the hydroxylation of heterocycles and the oxidation of aldehydes to their corresponding carboxylic acids. Different animal species have different complements of AOX genes. The two extremes are represented in humans and rodents; whereas the human genome contains a single active gene (AOX1), those of rodents, such as mice, are endowed with four genes (Aox1-4), clustering on the same chromosome, each encoding a functionally distinct AOX enzyme. It still remains enigmatic why some species have numerous AOX enzymes, whereas others harbor only one functional enzyme. At present, little is known about the physiological relevance of AOX enzymes in humans and their additional forms in other mammals. These enzymes are expressed in the liver and play an important role in the metabolisms of drugs and other xenobiotics. In this review, we discuss the expression, tissue-specific roles, and substrate specificities of the different mammalian AOX enzymes and highlight insights into their physiological roles.
doi: 10.1074/jbc.REV119.007741

Fast Prototyping Microfluidics: Integrating Droplet Digital Lamp for Absolute Quantification of Cancer Biomarkers - Oliveira, B.; B. Veigas; Fernandes, A. R.; Aguas, H.; R. Martins; E. Fortunato; P.V. Baptista (Sensors (Basel), 2020)
Microfluidic (MF) advancements have been leveraged toward the development of state-of-the-art platforms for molecular diagnostics, where isothermal amplification schemes allow for further simplification of DNA detection and quantification protocols. The MF integration with loop-mediated isothermal amplification (LAMP) is today the focus of a new generation of chip-based devices for molecular detection, aiming at fast and automated nucleic acid analysis. Here, we combined MF with droplet digital LAMP (ddLAMP) on an all-in-one device that allows for droplet generation, target amplification, and absolute quantification. This multilayer 3D chip was developed in less than 30 minutes by using a low-cost and extremely adaptable production process that exploits direct laser writing technology in "Shrinky-dinks" polystyrene sheets. ddLAMP and target quantification were performed directly on-chip, showing a high correlation between target concentration and positive droplet score. We validated this integrated chip via the amplification of targets ranging from five to 500,000 copies/reaction. Furthermore, on-chip amplification was performed in a 10 microL volume, attaining a limit of detection of five copies/microL under 60 min. This technology was applied to quantify a cancer biomarker, c-MYC, but it can be further extended to any other disease biomarker.
Oliveira, BeatrizVeigas, Bruno Fernandes, Alexandra R Aguas, Hugo Martins, Rodrigo Fortunato, Elvira Baptista, Pedro Viana eng UID/CTM/50025/2019; UID/Multi/04378/2019; Inn-INDIGO/0002/2015; PTDC/BTM-SAL/31201/2017; SFRH/BPD/124311/2016/Fundacao para a Ciencia e Tecnologia Switzerland Sensors (Basel). 2020 Mar 14;20(6). pii: s20061624. doi: 10.3390/s20061624.
Keywords: *Biosensing Techniques; Biomarkers, Tumor/genetics/*isolation & purification; digital amplification; DNA, Neoplasm/genetics/*isolation & purification; Humans; integrated microfluidics device; lab-on-a-chip; Lab-On-A-Chip Devices; Limit of Detection; loop-mediated isothermal amplification; Microfluidics/methods; Molecular Diagnostic Techniques/methods; Neoplasms/*diagnosis/genetics/pathology; Nucleic Acid Amplification Techniques/methods; oncogenes; Pathology, Molecular/methods

Flexible random lasers in dye-doped bio-degradable cellulose nanocrystalline needles - Germano, Gleice C. M.; Machado, Yan D. R.; Martinho, Lucas; Fernandes, Susete N.; Costa, Antonio Mario L. M.; Pecoraro, Edison; Gomes, Anderson S. L.; Carvalho, Isabel C. S. (Journal of the Optical Society of America BJournal of the Optical Society of America B, 2020)
In this work, we developed and investigated a random laser based on rhodamine6G (Rh6G) in ethylene glycol (EG) solution with varying cellulose nanocrystalline (CNC) needles as scatterers in the lasing media. Besides the suspension-in-cuvette scheme, an alternative configuration was also employed: a dye-CNC flexible self-supported thick-film (70 µm) random laser made by drop casting of the ${\rm CNCs}+{\rm Rh6G}+{\rm hydroxypropyl}$CNCs+Rh6G+hydroxypropyl cellulose suspension. In relation to conventional scatterers, the biodegradable cellulose nanocompounds showed a comparable reduction in both the spectral full width at half-maximum and the energy threshold values, with an optimal concentration of 5 mg [CNC]/ml[EG] in suspension. Its performance was also compared with other cellulose-based random lasers, presenting advantages for some parameters. The flexible film configuration showed similar results, but contained 10% less Rh6G than the suspension.
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Keywords: Optical feedback; Optical Fibers; Random lasers; Rare earth doped glass; Scattering media; Surface plasmons

Functional Implications of Second-Shell Basic Residues for dUTPase DR2231 Enzymatic Specificity - Vidossich, Pietro; Castañeda Moreno, Luis Eduardo; Mota, Cristiano; de Sanctis, Daniele; Miscione, Gian Pietro; De Vivo, Marco (ACS CatalysisACS Catalysis, 2020)
Nucleotide-processing enzymes are key players in biological processes. They often operate through high substrate specificity for catalysis. How such specificity is achieved is unclear. Here, we dealt with this question by investigating all-α dimeric deoxyuridine triphosphate nucleotidohydrolases (dUTPases). Typically, these dUTPases hydrolyze either dUTP or deoxyuridine diphosphate (dUDP) substrates. However, the dUTPase enzyme DR2231 from Deinococcus radiodurans selectively hydrolyzes dUTP only, and not dUDP. By means of extended classical molecular dynamics simulations and quantum chemical calculations, we show that DR2231 achieves this specificity for dUTP via second-shell basic residues that, together with the two catalytic magnesium ions, contribute to properly orienting the γ-phosphate of dUTP in a prereactive state. This allows a nucleophilic water to be correctly placed and activated in order to perform substrate hydrolysis. We show that this enzymatic mechanism is not viable when dUDP is bound to DR2231. Importantly, in several other dUTPases capable of hydrolyzing either dUTP or dUDP, we detected that active site second-shell basic residues are more in number, anchoring the β-phosphate of the nucleotide substrate too, in contrast to what is observed in DR2231. Thus, strategically located basic second-shell residues mediate precise reactant positioning at the catalytic site, determining substrate specificity in dUTPases and possibly in other structurally similar nucleotide-processing metalloenzymes.Nucleotide-processing enzymes are key players in biological processes. They often operate through high substrate specificity for catalysis. How such specificity is achieved is unclear. Here, we dealt with this question by investigating all-α dimeric deoxyuridine triphosphate nucleotidohydrolases (dUTPases). Typically, these dUTPases hydrolyze either dUTP or deoxyuridine diphosphate (dUDP) substrates. However, the dUTPase enzyme DR2231 from Deinococcus radiodurans selectively hydrolyzes dUTP only, and not dUDP. By means of extended classical molecular dynamics simulations and quantum chemical calculations, we show that DR2231 achieves this specificity for dUTP via second-shell basic residues that, together with the two catalytic magnesium ions, contribute to properly orienting the γ-phosphate of dUTP in a prereactive state. This allows a nucleophilic water to be correctly placed and activated in order to perform substrate hydrolysis. We show that this enzymatic mechanism is not viable when dUDP is bound to DR2231. Importantly, in several other dUTPases capable of hydrolyzing either dUTP or dUDP, we detected that active site second-shell basic residues are more in number, anchoring the β-phosphate of the nucleotide substrate too, in contrast to what is observed in DR2231. Thus, strategically located basic second-shell residues mediate precise reactant positioning at the catalytic site, determining substrate specificity in dUTPases and possibly in other structurally similar nucleotide-processing metalloenzymes.
doi: 10.1021/acscatal.0c04148

Gene Therapy in Cancer Treatment: Why Go Nano? - Roma-Rodrigues, C.; Rivas-Garcia, L.; P.V. Baptista; Fernandes, A. R. (Pharmaceutics, 2020)
The proposal of gene therapy to tackle cancer development has been instrumental for the development of novel approaches and strategies to fight this disease, but the efficacy of the proposed strategies has still fallen short of delivering the full potential of gene therapy in the clinic. Despite the plethora of gene modulation approaches, e.g., gene silencing, antisense therapy, RNA interference, gene and genome editing, finding a way to efficiently deliver these effectors to the desired cell and tissue has been a challenge. Nanomedicine has put forward several innovative platforms to overcome this obstacle. Most of these platforms rely on the application of nanoscale structures, with particular focus on nanoparticles. Herein, we review the current trends on the use of nanoparticles designed for cancer gene therapy, including inorganic, organic, or biological (e.g., exosomes) variants, in clinical development and their progress towards clinical applications.
Roma-Rodrigues, CatarinaRivas-Garcia, Lorenzo Baptista, Pedro V Fernandes, Alexandra R eng Inn-Indigo 00002/2015 RA Detect/Fundacao para a Ciencia e a Tecnologia UID/Multi/04378/2019/Fundacao para a Ciencia e a Tecnologia SFRH/BPD/124612/2016/Fundacao para a Ciencia e a Tecnologia Review Switzerland Pharmaceutics. 2020 Mar 5;12(3). pii: pharmaceutics12030233. doi: 10.3390/pharmaceutics12030233.
Keywords: gene delivery; Gene therapy; Nanomedicine; nanoparticles; Tumor microenvironment

Gold Nanoparticles for Vectorization of Nucleic Acids for Cancer Therapeutics - Ferreira, D.; Fontinha, D.; Martins, C.; Pires, D.; Fernandes, A. R.; P.V. Baptista (Molecules, 2020)
Cancer remains a complex medical challenge and one of the leading causes of death worldwide. Nanomedicines have been proposed as innovative platforms to tackle these complex diseases, where the combination of several treatment strategies might enhance therapy success. Among these nanomedicines, nanoparticle mediated delivery of nucleic acids has been put forward as key instrument to modulate gene expression, be it targeted gene silencing, interference RNA mechanisms and/or gene edition. These novel delivery systems have strongly relied on nanoparticles and, in particular, gold nanoparticles (AuNPs) have paved the way for efficient delivery systems due to the possibility to fine-tune their size, shape and surface properties, coupled to the ease of functionalization with different biomolecules. Herein, we shall address the different molecular tools for modulation of expression of oncogenes and tumor suppressor genes and discuss the state-of-the-art of AuNP functionalization for nucleic acid delivery both in vitro and in vivo models. Furthermore, we shall highlight the clinical applications of these spherical AuNP based conjugates for gene delivery, current challenges, and future perspectives in nanomedicine.
Ferreira, DanielaFontinha, David Martins, Catarina Pires, David Fernandes, Alexandra R Baptista, Pedro V eng UIDB/04378/2020; M-ERA-NET2/0008/2016/Fundacao para a Ciencia e a Tecnologia Review Switzerland Molecules. 2020 Jul 31;25(15). pii: molecules25153489. doi: 10.3390/molecules25153489.
Keywords: gene editing; Gene silencing; Gold nanoparticles; Nanomedicine; therapeutic nucleic acid

Hyperthermia Induced by Gold Nanoparticles and Visible Light Photothermy Combined with Chemotherapy to Tackle Doxorubicin Sensitive and Resistant Colorectal Tumor 3D Spheroids - Roma-Rodrigues, C.; Pombo, I.; Fernandes, A. R.; P.V. Baptista (Int J Mol Sci, 2020)
Current cancer therapies are frequently ineffective and associated with severe side effects and with acquired cancer drug resistance. The development of effective therapies has been hampered by poor correlations between pre-clinical and clinical outcomes. Cancer cell-derived spheroids are three-dimensional (3D) structures that mimic layers of tumors in terms of oxygen and nutrient and drug resistance gradients. Gold nanoparticles (AuNP) are promising therapeutic agents which permit diminishing the emergence of secondary effects and increase therapeutic efficacy. In this work, 3D spheroids of Doxorubicin (Dox)-sensitive and -resistant colorectal carcinoma cell lines (HCT116 and HCT116-DoxR, respectively) were used to infer the potential of the combination of chemotherapy and Au-nanoparticle photothermy in the visible (green laser of 532 nm) to tackle drug resistance in cancer cells. Cell viability analysis of 3D tumor spheroids suggested that AuNPs induce cell death in the deeper layers of spheroids, further potentiated by laser irradiation. The penetration of Dox and earlier spheroid disaggregation is potentiated in combinatorial therapy with Dox, AuNP functionalized with polyethylene glycol (AuNP@PEG) and irradiation. The time point of Dox administration and irradiation showed to be important for spheroids destabilization. In HCT116-sensitive spheroids, pre-irradiation induced earlier disintegration of the 3D structure, while in HCT116 Dox-resistant spheroids, the loss of spheroid stability occurred almost instantly in post-irradiated spheroids, even with lower Dox concentrations. These results point towards the application of new strategies for cancer therapeutics, reducing side effects and resistance acquisition.
Roma-Rodrigues, CatarinaPombo, Ines Fernandes, Alexandra R Baptista, Pedro V eng UIDB/04378/2020/Ministerio da Ciencia, Tecnologia e Ensino Superior SFRH/BPD/124612/2016/Ministerio da Ciencia, Tecnologia e Ensino Superior Switzerland Int J Mol Sci. 2020 Oct 28;21(21). pii: ijms21218017. doi: 10.3390/ijms21218017.
Keywords: 3D spheroids; colorectal cancer; doxorubicin resistance; Gold nanoparticles; photothermy

Improving the Anti-inflammatory Response via Gold Nanoparticle Vectorization of CO-Releasing Molecules - Alexandra R. Fernandes; Mendonça-Martins, Inês; Santos, Marino F. A.; Luís R. Raposo; Rita Mendes; Marques, Joana; Romão, Carlos C.; Romão, Maria João; Santos-Silva, Teresa; Pedro V. Baptista (ACS Biomaterials Science & EngineeringACS Biomaterials Science & Engineering, 2020)
CO-releasing molecules (CORMs) have been widely studied for their anti-inflammatory, antiapoptotic, and antiproliferative effects. CORM-3 is a water-soluble Ru-based metal carbonyl complex, which metallates serum proteins and readily releases CO in biological media. In this work, we evaluated the anti-inflammatory and wound-healing effects of gold nanoparticles–CORM-3 conjugates, AuNPs@PEG@BSA·Ru(CO)x, exploring its use as an efficient CO carrier. Our results suggest that the nanoformulation was capable of inducing a more pronounced cell effect, at the anti-inflammatory level and a faster tissue repair, probably derived from a rapid cell uptake of the nanoformulation that results in the increase of CO inside the cell.CO-releasing molecules (CORMs) have been widely studied for their anti-inflammatory, antiapoptotic, and antiproliferative effects. CORM-3 is a water-soluble Ru-based metal carbonyl complex, which metallates serum proteins and readily releases CO in biological media. In this work, we evaluated the anti-inflammatory and wound-healing effects of gold nanoparticles–CORM-3 conjugates, AuNPs@PEG@BSA·Ru(CO)x, exploring its use as an efficient CO carrier. Our results suggest that the nanoformulation was capable of inducing a more pronounced cell effect, at the anti-inflammatory level and a faster tissue repair, probably derived from a rapid cell uptake of the nanoformulation that results in the increase of CO inside the cell.
doi: 10.1021/acsbiomaterials.9b01936

Improving the Anti-inflammatory Response via Gold Nanoparticle Vectorization of CO-Releasing Molecules - Alexandra R. Fernandes; Mendonça-Martins, Inês; Santos, Marino F. A.; Luís R. Raposo; Rita Mendes; Marques, Joana; Romão, Carlos C.; Romão, Maria João; Santos-Silva, Teresa; Pedro V. Baptista (ACS Biomaterials Science & Engineering, 2020)
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In vitro antiproliferative effect of vanadium complexes bearing 8-hydroxyquinoline-based ligands - the substituent effect - Choroba, K.; Raposo, L. R.; Palion-Gazda, J.; Malicka, E.; Erfurt, K.; Machura, B.; Fernandes, A. R. (Dalton Trans, 2020)
This is the first comprehensive study demonstrating the antiproliferative effect of vanadium complexes bearing 8-hydroxyquinoline (quinH) ligands, including the parent and -CH3 (Me), -NO2, -Cl and -I substituted ligands, on HCT116 and A2780 cancer cell lines. To determine the structure-cytotoxicity relationships seven six-coordinate oxovanadium(v) complexes [VO(OMe)(5,7-(Me)2-quin)2] (1), [VO(OMe)(5,7-Cl2-quin)2] (2), [VO(OMe)(5,7-Cl,I-quin)2] (3), [VO(OMe)(5,7-I2-quin)2] (4), [VO(OMe)(5-NO2-quin)2] (5), [VO(OMe)(5-Cl-quin)2] (6), and [VO(OMe)(quin)2] (7) were investigated. The cytotoxicity of 8-hydroxyquinoline oxovanadium(v) complexes is higher in the A2780 cell line (lower IC50) than that observed for the widely used chemotherapeutic agent, cisplatin, while displaying low cytotoxicity for normal human primary fibroblasts. Substituents introduced into the 8-hydroxyquinoline backbone reduced the antiproliferative effect of the vanadium complexes, and the complexes with the ligand substituted only in the 5 position (5 and 6) were more cytotoxic than those with substituents in the 5,7 positions of the quin backbone (1-4). Depending on the substituent type, the cytotoxicity of 1-4 followed the trend: -Cl > -CH3 > -I. Incubation of A2780 cancer cells with IC50 concentrations of complexes 5, 6 and 7 promoted cellular detachment, possibly through membrane destabilization, and triggered apoptosis and necrosis. ROS production might be responsible for the cell death mechanism observed particularly in the A2780 cells exposed to complexes 5 and 6.
Choroba, KatarzynaRaposo, Luis R Palion-Gazda, Joanna Malicka, Ewa Erfurt, Karol Machura, Barbara Fernandes, Alexandra R eng England Dalton Trans. 2020 May 26;49(20):6596-6606. doi: 10.1039/d0dt01017k.

The Intracellular Number of Magnetic Nanoparticles Modulates the Apoptotic Death Pathway after Magnetic Hyperthermia Treatment - Beola, L.; Asin, L.; Roma-Rodrigues, C.; Fernandez-Afonso, Y.; Fratila, R. M.; Serantes, D.; Ruta, S.; Chantrell, R. W.; Fernandes, A. R.; P.V. Baptista; J.M. de la Fuente; Grazu, V.; et al. (ACS Appl Mater Interfaces, 2020)
Magnetic hyperthermia is a cancer treatment based on the exposure of magnetic nanoparticles to an alternating magnetic field in order to generate local heat. In this work, 3D cell culture models were prepared to observe the effect that a different number of internalized particles had on the mechanisms of cell death triggered upon the magnetic hyperthermia treatment. Macrophages were selected by their high capacity to uptake nanoparticles. Intracellular nanoparticle concentrations up to 7.5 pg Fe/cell were measured both by elemental analysis and magnetic characterization techniques. Cell viability after the magnetic hyperthermia treatment was decreased to <25% for intracellular iron contents above 1 pg per cell. Theoretical calculations of the intracellular thermal effects that occurred during the alternating magnetic field application indicated a very low increase in the global cell temperature. Different apoptotic routes were triggered depending on the number of internalized particles. At low intracellular magnetic nanoparticle amounts (below 1 pg Fe/cell), the intrinsic route was the main mechanism to induce apoptosis, as observed by the high Bax/Bcl-2 mRNA ratio and low caspase-8 activity. In contrast, at higher concentrations of internalized magnetic nanoparticles (1-7.5 pg Fe/cell), the extrinsic route was observed through the increased activity of caspase-8. Nevertheless, both mechanisms may coexist at intermediate iron concentrations. Knowledge on the different mechanisms of cell death triggered after the magnetic hyperthermia treatment is fundamental to understand the biological events activated by this procedure and their role in its effectiveness.
Beola, LilianneAsin, Laura Roma-Rodrigues, Catarina Fernandez-Afonso, Yilian Fratila, Raluca M Serantes, David Ruta, Sergiu Chantrell, Roy W Fernandes, Alexandra R Baptista, Pedro V de la Fuente, Jesus M Grazu, Valeria Gutierrez, Lucia eng ACS Appl Mater Interfaces. 2020 Sep 30;12(39):43474-43487. doi: 10.1021/acsami.0c12900. Epub 2020 Sep 17.
Keywords: 3D cell culture; Apoptosis; cell death pathways; iron oxides; Macrophages; Magnetic hyperthermia; nanoparticle uptake

Ionically Modified Cellulose Nanocrystal Self-Assembled Films with a Mesoporous Twisted Superstructure: Polarizability and Application in Ion-Gated Transistors - Grey, Paul; Fernandes, Susete N.; Gaspar, Diana; Deuermeier, Jonas; Rodrigo Martins; Elvira Fortunato; Godinho, Maria H; Pereira, Luis (ACS Applied Electronic MaterialsACS Applied Electronic Materials, 2020)
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doi: 10.1021/acsaelm.9b00652

Lazy State Determination: More Concurrency for Contending Linearizable Transactions - Vale, T. M.; Leitão, J.; Preguiça, N.; Rodrigues, R; Dias, R.J.; Lourenço, J. M. (FCT-NOVA, 2020)

Light Irradiation of Gold Nanoparticles Toward Advanced Cancer Therapeutics - Amendoeira, Ana; García, Lorenzo Rivas; Alexandra R. Fernandes; Pedro V. Baptista (2020)
Abstract Cancer is one of the leading causes of death in the world. To challenge this epidemic, there are growing demands for the development of new advanced and targeted therapeutics capable of effectively tackling cancer cells with improved selectivity. Nanomedicine has put forward several innovative therapeutics toward improving therapeutic efficacy while decreasing the deleterious side effects of current chemotherapy. Multifunctional gold nanoparticles (AuNPs) have been at the core of a plethora of advanced therapeutic strategies that provide selective targeting with their unique optical properties, capable to interact with the light of specific wavelength to deliver therapy with tremendous spatiotemporal precision. AuNPs have been exploited as photodynamic and photothermal therapeutic agents alone or in combination with other cancer treatment modalities with other cancer applications. Due to their exceptional physicochemical properties, they have been proven efficacious allies for photodynamic therapy and for photothermal therapy regimens. Herein, the rapidly progressing literature related to the use of these promising strategies against cancer is discussed, highlighting their possible future clinical translation.
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Magnetic Precipitation: A New Platform for Protein Purification - Raquel dos Santos; Iria, Inês; Manuel, Ana M; Leandro, Ana P; Madeira, Catarina A C; Goncalves, Joao; Ana Luísa Carvalho; Roque, Ana Cecília (Biotechnology JournalBiotechnology Journal, 2020)
One of the trends in downstream processing comprises the use of ?anything-but-chromatography? methods to overcome the current downfalls of standard packed-bed chromatography. Precipitation and magnetic separation are two techniques already proven to accomplish protein purification from complex media, yet never used in synergy. With the aim to capture antibodies directly from crude extracts, a new approach combining precipitation and magnetic separation was developed and named as affinity magnetic precipitation. A precipitation screening, based on the Hofmeister series, and a commercial precipitation kit were tested with affinity magnetic particles to assess the best condition for antibody capture from human serum plasma and clarified cell supernatant. The best conditions were obtained when using PEG3350 as precipitant at 4°C for 1h, reaching 80% purity and 50% recovery of polyclonal antibodies from plasma, and 99% purity with 97% recovery yield of anti-TNFα mAb from cell supernatants. These results show that the synergetic use of precipitation and magnetic separation can represent an alternative for the efficient capture of antibodies. This article is protected by copyright. All rights reserved
doi: 10.1002/biot.202000151
Keywords: affinity magnetic precipitation; antibody purification; downstream; Magnetic nanoparticles; method development

Multitask ATPases (NBDs) of bacterial ABC importers type I and their interspecies exchangeability - Leisico, Francisco; Lia M. Godinho; Gonçalves, Inês C.; Silva, Sara P.; Carneiro, Bruno; Romão, Maria J.; Santos-Silva, Teresa; Isabel de Sá-Nogueira (2020)
ATP-binding cassette (ABC) type I importers are widespread in bacteria and play a crucial role in its survival and pathogenesis. They share the same modular architecture comprising two intracellular nucleotide-binding domains (NBDs), two transmembrane domains (TMDs) and a substrate-binding protein. The NBDs bind and hydrolyze ATP, thereby generating conformational changes that are coupled to the TMDs and lead to substrate translocation. A group of multitask NBDs that are able to serve as the cellular motor for multiple sugar importers was recently discovered. To understand why some ABC importers share energy-coupling components, we used the MsmX ATPase from Bacillus subtilis as a model for biological and structural studies. Here we report the first examples of functional hybrid interspecies ABC type I importers in which the NBDs could be exchanged. Furthermore, the first crystal structure of an assigned multitask NBD provides a framework to understand the molecular basis of the broader specificity of interaction with the TMDs.
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Nano-in-Micro Sildenafil Dry Powder Formulations for the Treatment of Pulmonary Arterial Hypertension Disorders: The Synergic Effect of POxylated Polyurea Dendrimers, PLGA, and Cholesterol - Rita B. Restani; Rita F. Pires; Pedro V. Baptista; Alexandra R. Fernandes; Casimiro, Teresa; Vasco D.B. Bonifácio; Aguiar-Ricardo, Ana (2020)
Abstract POXylated polyurea dendrimer nanoparticles (PUREG4OOx48) are loaded with sildenafil (SDF) by a supercritical carbon dioxide–assisted (scCO2) impregnation. Further supercritical CO2-assisted spray drying (SASD) leads to hybrid nano-in-micro dry powder formulations that are investigated aiming at efficient pulmonary delivery of SDF in pulmonary arterial hypertension treatment. This is the first report of the production of poly(D,L-lactide-co-glycolide)-cholesterol (PLGA-Chol) microparticles processed by SASD. The optimized formulation of nano-in-microparticles is composed of PLGA, Chol, and PUREG4OOx48, loaded with SDF solutions in a 77:23 ratio (PLGA-Chol:dendrimer, w/w). The dry powders are fully characterized and found to be highly biodegradable and biocompatible, and the SDF release profile evaluates under different pH values. The median mass average diameter (MMAD) of the nano-in-micro systems varies between 2.57 and 5 µm and the fine particle fraction (FPF) between 36% and 29% for PUREG4OMeOx48[PLGA-Chol] and PUREG4OEtOx48[PLGA-Chol], respectively. The data validate the potential use of these new formulations in inhalation therapy. In vitro studies are also carried out in order to evaluate the effect of the free drug in cell viability and formulations cytotoxicity.
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New copper(I) complexes selective for prostate cancer cells - Machado, J. F.; Sequeira, D.; Marques, F.; Piedade, M. F. M.; Villa de Brito, M. J.; Helena Garcia, M.; Fernandes, A. R.; Morais, T. S. (Dalton Trans, 2020)
A new family of eighteen Cu(i) complexes of the general formula [Cu(PP)(LL)][BF4], where PP is a phosphane ligand and LL represents an N,O-heteroaromatic bidentate ligand, has been synthesized and fully characterized by classical analytical and spectroscopic methods. Five complexes of this series were also characterized by single crystal X-ray diffraction studies. The cytotoxicity of all compounds was evaluated in breast (MCF7) and prostate (LNCap) human cancer cells and in a normal prostate cell line (RWPE). In general, all compounds showed higher cytotoxicity for the prostate cancer cells than for the breast cells, with IC50 values in the range 0.2-2 muM after 24 h of treatment. The most cytotoxic compound, [Cu(dppe)(2-ap)][BF4] (16), where dppe = 1,2-bis(diphenylphosphano) ethane and 2-ap = 2-acetylpyridine, showed a high level of cellular internalization, generation of intracellular ROS and activation of the cell death mechanism via apoptosis/necrosis. Owing to its high cytotoxic activity for LNCap cells, being 70-fold higher than that for normal prostate cells (RWPE), complex (16) was found to be the most promising for further research in prostate cancer models.
Machado, Joao FrancoSequeira, Diogo Marques, Fernanda Piedade, M Fatima M Villa de Brito, Maria J Helena Garcia, M Fernandes, Alexandra R Morais, Tania S eng England Dalton Trans. 2020 Sep 15;49(35):12273-12286. doi: 10.1039/d0dt02157a.

Photonic composite materials from cellulose nanorods and clay nanolayers - Trindade, Ana C.; Carreto, Miguel; Helgesen, Geir; Knudsen, Kenneth D.; Puchtler, Florian; Breu, Josef; Fernandes, Susete; Godinho, Maria Helena; Fossum, Jon Otto (2020)
Cellulose nano crystals (CNCs) are promising materials for energy efficient buildings related to the control of reflectivity and heat absorption/reflection of light. In this sense it is important to improve CNCs films fire retardant properties, which can be achieved by adding clays. Cellulose nanocrystals (CNCs) and nanolayers obtained from Sodium Fluorohectorite (NaFh) synthetic clay are both known to form liquid crystalline phases in aqueous suspensions. CNCs form cholesteric phases, which structure is preserved after water evaporation, while dry NaFh nanolayers aligned films collapse. In this initial work, it is shown that CNCs are compatible with NaFh clay. We demonstrate that the liquid crystalline phase of CNCs in water is not destroyed by the presence of NaFh nanolayers. The NaFh nanolayers act as planar anchoring surfaces to the cellulose nanorods and, after evaporation of the water coloured films are obtained. The precursor solutions and the photonic films were investigated by Describe several techniques.
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Playing the blues, the greens and the reds with cellulose-based structural colours - Chagas, Ricardo; Silva, Pedro E. S.; Fernandes, Susete N.; Žumer, Slobodan; Godinho, Maria Helena (Faraday Discussions, 2020)
Structural vivid colours can arise from the interference of light reflected from structures exhibiting periodicity on scales in the range of visible wavelengths. This effect is observed with light reflected from cell-walls of some plants and exoskeletons of certain insects. Sometimes the colour sequence observed for these structures consists of nearly circular concentric rings that vary in colour from Red, Orange, Yellow, Green, Cyan to Blue, from the periphery to the centre, similarly to the colour scheme sequence observed for the rainbow (ROYGB). The sequence of colours has been found for solid films obtained from droplets of aqueous cellulose nanocrystals (CNCs) suspensions and attributed to a “coffee ring” effect. In this work, coloured lyotropic solutions and solid films obtained from a cellulose derivative in the presence of trifluoroacetic acid (TFA), which acts as a “reactive solvent”, are revisited. The systems were investigated with spectroscopy, using circularly and linearly polarised light, coupled with a polarised optical microscope (POM) and scanning electron microscopy (SEM). The lyotropic cholesteric liquid crystalline solutions were confined in capillaries to simplify 1D molecular diffusion along the capillary where an unexpected sequence of the structural colours was observed. The development and reappearance of the sequence of vivid colours seem consistent with the reaction–diffusion of the “reactive solvent” in the presence of the cellulosic chains. The strong TFA acts as an auto-catalyst for the chemical reaction between TFA and the hydroxyl groups, existing along the cellulosic chain, and diffuses to the top and bottom along the capillaries, carrying dissolved cellulosic chains. Uncovering the precise mechanism of colour sequence and evolution over time in cellulosic lyotropic solutions has important implications for future optical/sensors applications and for the understanding of the development of cellulose-based structures in nature.
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Singularities of Pyogenic Streptococcal Biofilms – From Formation to Health Implication - Alves-Barroco, Cinthia; Paquete-Ferreira, João; Santos-Silva, Teresa; Alexandra R. Fernandes (2020)
Biofilms are generally defined as communities of cells involved in a self-produced extracellular matrix adhered to a surface. In biofilms, the bacteria are less sensitive to host defense mechanisms and antimicrobial agents, due to multiple strategies, that involve modulation of gene expression, controlled metabolic rate, intercellular communication, composition, and 3D architecture of the extracellular matrix. These factors play a key role in streptococci pathogenesis, contributing to therapy failure and promoting persistent infections. The species of the pyogenic group together with Streptococcus pneumoniae are the major pathogens belonging the genus Streptococcus, and its biofilm growth has been investigated, but insights in the genetic origin of biofilm formation are limited. This review summarizes pyogenic streptococci biofilms with details on constitution, formation, and virulence factors associated with formation.
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Keywords: Biofilm,antibiotic resistance,Virulence Factors,Quorum-Sensing 2,Streptococcal

Size-Dependent Biological Activities of Fluorescent Organosilane-Modified Zinc Oxide Nanoparticles - Busila, M.; Tabacaru, A.; Mussat, V.; Vasile, B. S.; Neasu, I. A.; Pinheiro, T.; Roma-Rodrigues, C.; P.V. Baptista; Fernandes, A. R.; Matos, A. P.; Marques, F. (J Biomed Nanotechnol, 2020)
Surface modification of zinc oxide nanoparticles (ZnO NPs) is a strategy to tune their biocompatibility. Herein we report on the synthesis of a series of fluorescent ZnO NPs modified with 2-10% (3-glycidyloxypropyl)trimethoxysilane (GPTMS) to investigate the fluorescence properties and to explore their applications in microbiology and biomedicine. The obtained ZnO NPs were characterized by X-ray diffraction (XRD), high resolution transmission electron microscopy (HRTEM) and Fourier transform infrared spectroscopy (FTIR). Size reduction occurred from ca. 13 nm in unmodified ZnO to 3-4 nm in silane-modified samples and fluorescence spectra showed size-dependent variation of the photoemission bands' intensity. The antibacterial and cytotoxic activities were investigated on Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacteria, and in ovarian (A2780) and prostate (PC3) cancer cells by tetrazolium/formazan-based methods. The antibacterial effect was higher for E. coli than S. aureus, while the cytotoxic activity was similar for both cancer cells and varied with the particle size. Cell death by apoptosis, and/or necrosis versus autophagy, were explored by flow cytometry using an Annexin V based-method and transmission electron microscopy (TEM). The main mechanism of ZnO NPs toxicity may involve the generation of reactive oxygen species (ROS) and the induction of apoptosis or autophagy. This work revealed the potential utility of GPTMS-modified ZnO NPs in the treatment of bacterial infection and cancer.
Busila, MarianaTabacaru, Aurel Mussat, Viorica Vasile, Bogdan Stefan Neasu, Ionela Andreea Pinheiro, Teresa Roma-Rodrigues, Catarina Baptista, Pedro V Fernandes, Alexandra R Matos, Antonio P Marques, Fernanda eng J Biomed Nanotechnol. 2020 Feb 1;16(2):137-152. doi: 10.1166/jbn.2020.2882.
Keywords: *Metal Nanoparticles; *Ovarian Neoplasms; Anti-Bacterial Agents; Cell Line, Tumor; Escherichia coli; Female; Humans; Microbial Sensitivity Tests; Organosilicon Compounds; Spectroscopy, Fourier Transform Infrared; Staphylococcus aureus; X-ray diffraction; Zinc oxide

SLMP53-1 interacts with wild-type and mutant p53 DNA-binding domain and reactivates multiple hotspot mutations - Gomes, Ana Sara; Ramos, Helena; Gomes, Sara; Loureiro, Joana B.; Soares, Joana; Barcherini, Valentina; Monti, Paola; Fronza, Gilberto; Oliveira, Carla; Domingues, Lucília; Bastos, Margarida; Dourado, Daniel F.A.R.; et al. (2020)
BackgroundHalf of human cancers harbour TP53 mutations that render p53 inactive as a tumor suppressor. As such, reactivation of mutant (mut)p53 through restoration of wild-type (wt)-like function represents one of the most promising therapeutic strategies in cancer treatment. Recently, we have reported the (S)-tryptophanol-derived oxazoloisoindolinone SLMP53-1 as a new reactivator of wt and mutp53 R280K with in vitro and in vivo p53-dependent antitumor activity. The present work aimed a mechanistic elucidation of mutp53 reactivation by SLMP53-1. Methods and results By cellular thermal shift assay (CETSA), it is shown that SLMP53-1 induces wt and mutp53 R280K thermal stabilization, which is indicative of intermolecular interactions with these proteins. Accordingly, in silico studies of wt and mutp53 R280K DNA-binding domain with SLMP53-1 unveiled that the compound binds at the interface of the p53 homodimer with the DNA minor groove. Additionally, using yeast and p53-null tumor cells ectopically expressing distinct highly prevalent mutp53, the ability of SLMP53-1 to reactivate multiple mutp53 is evidenced. Conclusions SLMP53-1 is a p53-activating agent with the ability to directly target wt and a set of hotspot mutp53. General Significance This work reinforces the encouraging application of SLMP53-1 in the personalized treatment of cancer patients harboring distinct p53 status.
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Keywords: Cancer; chemotherapy; Mutant; p53; Reactivator

Synthesis of new hetero-arylidene-9(10H)-anthrone derivatives and their biological evaluation - Roma-Rodrigues, C.; Malta, G.; Peixoto, D.; Ferreira, LM; P.V. Baptista; Fernandes, A. R.; Branco, P. S. (Bioorg Chem, 2020)
New hetero-arylidene-9(10H)-anthrone derivatives (1) were synthesized from reaction of 1,2-dimethyl-3-alkyl imidazolium salts (2) and 9-anthracenecarboxaldehyde. Ion exchange of the anion with dioctyl sulfosuccinate and lithium bis(trifluoromethanesulfonyl)imide led to the preparation of other derivatives. The antiproliferative effect of the compounds was evaluated in human ovarian (A2780) and colorectal (HCT116) carcinoma cell lines and in normal primary human fibroblasts. Compound 1 presented an antiproliferative effect related to the imidazolium pattern of substitution with compounds having a decyl group at the R-position (1c and 3c) showing the highest cytotoxic activities in all cell lines independently of the counter ion. Compounds 1b and 1c internalize A2780 cancer cells via a passive or an active transport, respectively, inducing A2780 cell death via an extrinsic apoptosis (1b) or intrinsic apoptosis and oncosis (1c). The localization of both compounds in the cytoplasm coupled to the absence of reactive oxygen species (ROS) induction suggest that the mechanisms of toxicity might be different than those of other anthracyclines currently used in chemotherapy.
Roma-Rodrigues, CatarinaMalta, Gabriela Peixoto, Daniela Ferreira, Luisa M Baptista, Pedro V Fernandes, Alexandra R Branco, Paula S eng Research Support, Non-U.S. Gov't Bioorg Chem. 2020 Jun;99:103849. doi: 10.1016/j.bioorg.2020.103849. Epub 2020 Apr 14.
Keywords: *Arylidene-anthrone derivatives; *Cell apoptosis; *Imidazolium chemistry; *Ovarian cancer; *Proliferation inhibition; competing financial interests or personal relationships that could have appeared; to influence the work reported in this paper.

Tackling Multidrug Resistance in Streptococci - From Novel Biotherapeutic Strategies to Nanomedicines - Alves-Barroco, C.; Rivas-Garcia, L.; Fernandes, A. R.; P.V. Baptista (Front Microbiol, 2020)
The pyogenic streptococci group includes pathogenic species for humans and other animals and has been associated with enduring morbidity and high mortality. The main reason for the treatment failure of streptococcal infections is the increased resistance to antibiotics. In recent years, infectious diseases caused by pyogenic streptococci resistant to multiple antibiotics have been raising with a significant impact to public health and veterinary industry. The rise of antibiotic-resistant streptococci has been associated to diverse mechanisms, such as efflux pumps and modifications of the antimicrobial target. Among streptococci, antibiotic resistance emerges from previously sensitive populations as result of horizontal gene transfer or chromosomal point mutations due to excessive use of antimicrobials. Streptococci strains are also recognized as biofilm producers. The increased resistance of biofilms to antibiotics among streptococci promote persistent infection, which comprise circa 80% of microbial infections in humans. Therefore, to overcome drug resistance, new strategies, including new antibacterial and antibiofilm agents, have been studied. Interestingly, the use of systems based on nanoparticles have been applied to tackle infection and reduce the emergence of drug resistance. Herein, we present a synopsis of mechanisms associated to drug resistance in (pyogenic) streptococci and discuss some innovative strategies as alternative to conventional antibiotics, such as bacteriocins, bacteriophage, and phage lysins, and metal nanoparticles. We shall provide focused discussion on the advantages and limitations of agents considering application, efficacy and safety in the context of impact to the host and evolution of bacterial resistance.
Alves-Barroco, CinthiaRivas-Garcia, Lorenzo Fernandes, Alexandra R Baptista, Pedro Viana eng Review Switzerland Front Microbiol. 2020 Oct 6;11:579916. doi: 10.3389/fmicb.2020.579916. eCollection 2020.
Keywords: antimicrobial resistance; bacteriocins; bacteriophage; biofilms; Nanomedicine; nanoparticles; pyogenic streptococci

A Tale of Two Ends: Repurposing Metallic Compounds from Anti-Tumour Agents to Effective Antibacterial Activity - Alves Ferreira, D.; L, M. D. R. S. Martins; A, R. Fernandes; Martins, M (Antibiotics (Basel), 2020)
The rise in antibiotic resistance coupled with the gap in the discovery of active molecules has driven the need for more effective antimicrobials while focusing the attention into the repurpose of already existing drugs. Here, we evaluated the potential antibacterial activity of one cobalt and two zinc metallic compounds previously reported as having anticancer properties. Compounds were tested against a range of Gram-positive and -negative bacteria. The determination of the minimum inhibitory and bactericidal concentrations (MIC/MBC) of the drugs were used to assess their potential antibacterial activity and their effect on bacterial growth. Motility assays were conducted by exposing the bacteria to sub-MIC of each of the compounds. The effect of sub-MIC of the compounds on the membrane permeability was measured by ethidium bromide (EtBr) accumulation assay. Cell viability assays were performed in human cells. Compound TS262 was the most active against the range of bacteria tested. No effect was observed on the motility or accumulation of EtBr for any of the bacteria tested. Cell viability assays demonstrated that the compounds showed a decrease in cell viability at the MIC. These results are promising, and further studies on these compounds can lead to the development of new effective antimicrobials.
Alves Ferreira, DanielaM D R S Martins, Luisa R Fernandes, Alexandra Martins, Marta eng Department of Microbiology/Trinity College Dublin Postgraduate Research (1252) Studentship/Trinity College Dublin Unidade de Ciencias Biomoleculares Aplicadas - UCIBIO (UIDB/04378/2020)/Fundacao para a Ciencia e a Tecnologia Switzerland Antibiotics (Basel). 2020 Jun 11;9(6). pii: antibiotics9060321. doi: 10.3390/antibiotics9060321.
Keywords: antimicrobial resistance; drug repurposing; metallic compounds

Towards the mechanistic understanding of enzymatic CO2 reduction - Ana Rita Oliveira; Mota, Cristiano; Mourato, Cláudia; Domingos, Renato M.; Santos, Marino F. A.; Gesto, Diana; Guigliarelli, Bruno; Santos-Silva, Teresa; Romão, Maria João; Pereira, Inês Antunes Cardoso (ACS CatalysisACS Catalysis, 2020)
Reducing CO2 is a challenging chemical transformation that biology solves easily, with high efficiency and specificity. In particular, formate dehydrogenases are of great interest since they reduce CO2 to formate, a valuable chemical fuel and hydrogen storage compound. The metal-dependent formate dehydrogenases of prokaryotes can show high activity for CO2 reduction. Here, we report an expression system to produce recombinant W/Sec-FdhAB from Desulfovibrio vulgaris Hildenborough fully loaded with cofactors, its cata-lytic characterization and crystal structures in oxidised and reduced states. The enzyme has very high activi-ty for CO2 reduction and displays remarkable oxygen stability. The crystal structure of the formate-reduced enzyme shows Sec still coordinating the tungsten, supporting a mechanism of stable metal coordination during catalysis. Comparison of the oxidised and reduced structures shows significant changes close to the active site. The DvFdhAB is an excellent model for studying catalytic CO2 reduction and probing the mecha-nism of this conversion.Reducing CO2 is a challenging chemical transformation that biology solves easily, with high efficiency and specificity. In particular, formate dehydrogenases are of great interest since they reduce CO2 to formate, a valuable chemical fuel and hydrogen storage compound. The metal-dependent formate dehydrogenases of prokaryotes can show high activity for CO2 reduction. Here, we report an expression system to produce recombinant W/Sec-FdhAB from Desulfovibrio vulgaris Hildenborough fully loaded with cofactors, its cata-lytic characterization and crystal structures in oxidised and reduced states. The enzyme has very high activi-ty for CO2 reduction and displays remarkable oxygen stability. The crystal structure of the formate-reduced enzyme shows Sec still coordinating the tungsten, supporting a mechanism of stable metal coordination during catalysis. Comparison of the oxidised and reduced structures shows significant changes close to the active site. The DvFdhAB is an excellent model for studying catalytic CO2 reduction and probing the mecha-nism of this conversion.
doi: 10.1021/acscatal.0c00086

Zn(II) and Co(II) derivatives anchored with scorpionate precursor: Antiproliferative evaluation in human cancer cell lines - Das, K.; Datta, A.; Frontera, A.; Wen, Y. S.; Roma-Rodrigues, C.; Raposo, L. R.; Fernandes, A. R.; Hung, C. H. (J Inorg Biochem, 2020)
A 'scorpionate' type precursor [bdtbpza=bis(3,5-di-t-butylpyrazol-1-yl)acetate] has been employed to synthesize two mononuclear Zn(II) and Co(II) derivatives, namely [Zn(bdtbpza)2 (H2O)2].2.5CH3OH.2[(CH3)3C-C3H2N2-C(CH3)3] (1) and [Co(bdtbpza)2(CH3OH)4] (2) in good yield. Single crystal X-ray diffraction analysis reveals that in 1, the Zn(II) atom is tetrahedrally surrounded by a pair of Oacetate atoms of two bis(pyrazol-1-yl)acetate units and two water molecules; while in 2, the Co(II) atom shows an octahedral environment coordinating a pair of Oacetate atoms of two bis(pyrazol-1-yl)acetate units along with four methanol molecules. The EPR spectra of 2 recorded at 77 and 298K confirmed the tetragonal symmetry of the high spin Co(II). The DFT (Density functional theory) computation is in good agreement with the geometry proposed for compounds 1 and 2. Both the compounds display a high antiproliferative activity against HCT116 (colorectal carcinoma) and A2780 (ovarian carcinoma) cell lines compared to human normal dermal fibroblasts. In the case of A2780 cells, compounds 1 and 2 exhibit IC50 values that are similar to those described for cisplatin, a widely used chemotherapeutic drug. Exposure of A2780 cells to the IC50 concentration of each compound led to an increase of the number of apoptotic and autophagic cells. In the case of compound 1, the accumulation of intracellular ROS (Reactive oxygen species) is responsible for triggering A2780 cell death.
Das, KuheliDatta, Amitabha Frontera, Antonio Wen, Yuh-Sheng Roma-Rodrigues, Catarina Raposo, Luis R Fernandes, Alexandra R Hung, Chen-Hsiung eng Research Support, Non-U.S. Gov't J Inorg Biochem. 2020 Jan;202:110881. doi: 10.1016/j.jinorgbio.2019.110881. Epub 2019 Oct 23.
Keywords: *Antiproliferative; *Autophagy, ROS; *Cancer cell lines; *Co(II); *Zn(II)

Additive Manufacturing in health sector: Bio printing and Bio Manufacturing - Nuno Araújo; António Brandão Moniz (8th International Conference on Virtual and Networked Organizations Emergent Technologies and Tools - ViNOrg'20, 2020)

PRODUTECH SIF Project: its mobilized effect and main results and its contribution to accelerating digital transformation in the industry - V. Pacheco; Nuno Araújo; L. Rocha (8th International Conference on Virtual and Networked Organizations Emergent Technologies and Tools - ViNOrg'20, 2020)

Smart Additive Manufacturing: the path to digital chain - V. Pacheco; Nuno Araújo; L. Rocha (8th International Conference on Virtual and Networked Organizations Emergent Technologies and Tools - ViNOrg'20, 2020)

Anything but Conventional Chromatography Approaches in Bioseparation - Roque, A. C. A.; Pina, A. S.; A. M. Azevedo; Aires-Barros, R.; Jungbauer, A; Profio, G Di; Heng, JYY; Haigh, J; Ottens, M (Biotechnology Journal, 2020)

Assembly of peptidoglycan fragments – a synthetic challenge - F. Queda; G. Covas; S. R. Filipe*; Marques*, M M B (Pharmaceuticals, 2020)

Carbon threads sweat-based supercapacitors for electronic textiles - Nuno Lima; Baptista, Ana Catarina; Bruno M. Morais Faustino; Sofia Taborda; Marques, Ana; Ferreira, Isabel (Scientific Reports, 2020)
Flexible and stretchable energy-storage batteries and supercapacitors suitable for wearable electronics are at the forefront of the emerging field of intelligent textiles. In this context, the work here presented reports on the development of a symmetrical wire-based supercapacitor able to use the wearer’s sweat as the electrolyte. The inner and outer electrodes consists of a carbon-based thread functionalized with a conductive polymer (polypyrrole) which improves the electrochemical performances of the supercapacitor. The inner electrode is coated with electrospun cellulose acetate fibres, as the separator, and the outer electrode is twisted around it. The electrochemical performances of carbon-based supercapacitors were analyzed using a simulated sweat solution and displayed a specific capacitance of 2.3 F.g−1, an energy of 386.5 mWh.kg−1 and a power density of 46.4 kW.kg−1. Moreover, cycle stability and bendability studies were performed. Such energy conversion device has exhibited a stable electrochemical performance under mechanical deformation, over than 1000 cycles, which make it attractive for wearable electronics. Finally, four devices were tested by combining two supercapacitors in series with two in parallel demonstrating the ability to power a LED.

Carbon threads sweat-based supercapacitors for electronic textiles - Nuno Lima; Baptista, Ana C.; Bruno M. Morais Faustino; Sofia Taborda; Marques, Ana; Ferreira, Isabel (Scientific reports, 2020)

Carbon-Based Materials for the Development of Highly Dispersed Metal Catalysts: Towards Highly Performant Catalysts for Fine Chemical Synthesis - Pérez-Mayoral, E.; Matos, I.; Bernardo, M.; Ventura, M.; Fonseca, I.M. (Catalysts, 2020)

Conductive Electrospun Polyaniline/Polyvinylpyrrolidone Nanofibers: Electrical and Morphological Characterization of New Yarns for Electronic Textiles - Patrícia Perdigão; Bruno Faustino; Faria, Jaime; Canejo, João Paulo; Borges, João Borges; Ferreira, Isabel; Baptista, Ana Catarina (Fibers, 2020)
Advanced functionalities textiles embedding electronic fibers, yarns and fabrics are a demand for innovative smart cloths. Conductive electrospun membranes and yarns based on polyaniline/polyvinylpyrrolidone (PANI/PVP) were investigated using the chemical modification of PANI instead of using conventional coating processes as in-situ polymerization. PANI was synthesized from the aniline monomer and the influence of the oxidant-to-monomer ratio on electrical conductivity was studied. The optimized conductivity of pellets made with pressed PANI powders was 21 S·cm−1. Yarns were then prepared from the t-Boc-PANI/PVP electrospun membranes followed by PANI protonation to enhance their electrical properties. Using this methodology, electrospun membranes and yarns were produced with electrical conductivities of 1.7 × 10−2 and 4.1 × 10−4 S·cm−1.

Conductive electrospun polyaniline/polyvinylpyrrolidone nanofibers: Electrical and morphological characterization of new yarns for electronic textiles - Patrícia Perdigão; Bruno Miguel Faustino; Faria, Jaime; Canejo, João Paulo; Borges, João Paulo; Ferreira, Isabel; Baptista, Ana Catarina (Fibers, 2020)

Cyclodextrin solubilization and complexation of antiretroviral drug lopinavir: In silico prediction; Effects of derivatization, molar ratio and preparation method - Adeoye, Oluwatomide; Conceição, Jaime; Serra, Patrrícia A; da Silva, Andreia Bento; Duarte, Noélia; Guedes, Rita C.; Corvo, Marta C.; Aguiar-Ricardo, Ana; Jicsinszky, László; Casimiro, Teresa; Cabral-Marques, Helena (Carbohydrate Polymers, 2020)
Lopinavir (LPV) is currently used in combination with ritonavir for the clinical management of HIV infections due to its limited oral bioavailability. Herein, we report the application of an in silico method to study cyclodextrin (CyD) host-guest molecular interaction with LPV for the rational selection of the best CyD for developing a CyD based LPV delivery system. The predicted CyD, a (2-hydroxy)propyl-gamma derivative with high degree of substitution (HP17-γ-CyD) was synthesized and comparatively evaluated with γ-CyD and the commercially available HP-γ-CyD. All complexes were prepared by supercritical assisted spray drying (SASD) and co-evaporation (CoEva) at molar ratios (1:1 and 1:2); and afterwards fully characterized. Results indicate a higher LPV amorphization and solubilization ability of HP17-γ-CyD. The SASD processing technology also enhanced LPV solubilization and release from complexes. The application of in silico methodologies is a feasible approach for the rational and/or deductive development of CyD drug delivery systems.
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Demo of PolySense: How to Make Electrically Functional Textiles - P. Strohmeier; C. Honnet; H. Perner-Wilson; M. Teyssier; B. Fruchard; Baptista, Ana Catarina; J. Steimle (CHI Conference on Human Factors in Computing Systems, 2020)
We demonstrate a simple and accessible method for enhancing textiles with custom piezo-resistive properties. Based on in-situ polymerization, our method offers seamless integration at the material level, preserving a textile's haptic and mechanical properties. We demonstrate how to enhance a wide set of fabrics and yarns using only readily available tools. During each demo session, conference attendees may bring textile samples which will be polymerized in a shared batch. Attendees may keep these samples. While the polymerization is happening, attendees can inspect pre-made samples and explore how these might be integrated in functional circuits. Examples objects created using polymerization include rapid manufacturing of on-body interfaces, tie-dyed motion-capture clothing, and zippers that act as potentiometers.

Design of Artificial Enzymes Using the Metals of the Periodic Table - Moura J.J.G. (Memories of the Class of Sciences, Academia das Ciências de Lisboa, 2020)

Design of wave-optical structured substrates for ultra-thin perovskite solar cells - Haque, Sirazul; Alexandre, Miguel; Mendes, Manuel J.; Hugo Águas; Elvira Fortunato; Rodrigo Martins (Applied Materials Today, 2020)

Designed Metal-ATCUN Derivatives: Redox and Non-redox-Based Applications Relevant for Chemistry, Biology, and Medicine - B.K. Maiti; Govil, N.; Kundu, T.; Moura J.J.G. (iScience, 2020)

Designing silica xerogels containing RTIL for CO2 capture and CO2/CH4 separation: Influence of ILs anion, cation and cation side alkyl chain length and ramification - dos Santos, Leonardo M; Bernard, Franciele L.; Polesso, Bárbara B; Pinto, Ingrid S.; Frankenberg, Claudio C.; Corvo, Marta C.; Almeida, Pedro L; Eurico Cabrita; Menezes, Sonia; Einloft, Sandra (Journal of Environmental Management, 2020)
CO2 separation from natural gas is considered to be a crucial strategy to mitigate global warming problems, meet product specification, pipeline specs and other application specific requirements. Silica xerogels (SX) are considered to be potential materials for CO2 capture due to their high specific surface area. Thus, a series of silica xerogels functionalized with imidazolium, phosphonium, ammonium and pyridinium-based room-temperature ionic liquids (RTILs) were synthesized. The synthesized silica xerogels were characterized by NMR, helium pycnometry, DTA-TG, BET, SEM and TEM. CO2 sorption, reusability and CO2/CH4 selectivity were assessed by the pressure-decay technique. Silica xerogels containing IL demonstrated advantages compared to RTILs used as separation solvents in CO2 capture processes including higher CO2 sorption capacity and faster sorption/desorption. Using fluorinated anion for functionalization of silica xerogels leads to a higher affinity for CO2 over CH4. The best performance was obtained by SX- [bmim] [TF2N] (223.4 mg CO2/g mg/g at 298.15 K and 20 bar). Moreover, SX- [bmim] [TF2N] showed higher CO2 sorption capacity as compared to other reported sorbents. CO2 sorption and CO2/CH4 selectivity results were submitted to an analysis of variance and the means compared using Tukey's test (5%).
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Diverse biological roles of the tetrathiomolybdate anion - B.K. Maiti; Moura, J. J. G. (Coord Chem Rev, 2020)

Effect of Nanomaterials on Protein Amyloid Aggregation - K.Siposova, A.Musatov (International Conference - Condensed Matter Research at the IBR-2, 2020)

The effect of pH on Marinobacter hydrocarbonoclasticus denitrification pathway and nitrous oxide reductase - Carreira, C.; Nunes, R.F.; Mestre, O.; Moura, I.; Pauleta, S. R. (J Biol Inorg Chem, 2020)

Enhancement of CO2/N2 selectivity and CO2 uptake by tuning concentration and chemical structure of imidazolium-based ILs immobilized in mesoporous silica - Duczinski, Rafael; Polesso, Barbara B.; Bernard, Franciele L.; Ferrari, Henrique Z.; Almeida, Pedro L; Corvo, Marta C.; Cabrita, Eurico J.; Menezes, Sonia; Einloft, Sandra (Journal of Environmental Chemical Engineering, 2020)
Imidazolium-based ionic liquids (ILs) with different cation alkyl chain ([i-C5mim] or [C4mim]) and inorganic anions ([Cl−], [Tf2N−], [PF6−] and [DCA−]) were synthesized and immobilized in commercial mesoporous silica. The synthesized supported ILs (SILs) were characterized using NMR, FTIR, TGA, BET, SEM and TEM. CO2 sorption capacity, reusability and CO2/N2 selectivity were assessed by the pressure-decay technique. The effects of IL concentration, cation and anion chemical structure in CO2 sorption capacity and CO2/N2 separation performance were evaluated. Tests evidenced that the presence of branching on the cation alkyl side chain increases CO2/N2 selectivity. The immobilization of the IL [i-C5TPIm][Cl] on mesoporous silica in different concentrations (50, 20, 10 and 5 %) revealed that lower IL concentration results in higher CO2 sorption capacity. Immobilization of ILs containing fluorinated anions at low concentrations in the mesoporous silica support may promote the improvement of the CO2/N2 selectivity without interfering on CO2 sorption capacity of the original support. CO2 sorption capacity value shown by sample SIL-5 % - [i-C5TPIm][Tf2N] (79.50 ± 0.70 mg CO2  g-1) was close to the value obtained for the pristine mesoporous silica (81.70 ± 2.20 mg CO2 g-1) and the selectivity (4.30 ± 0.70) was more than twice of the one obtained for the support alone (2.32 ± 0.4). Recycle tests demonstrated that the ILs immobilized in mesoporous silica samples are stable, providing a new option to be used in CO2 capture processes.
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Fast Prototyping Microfluidics: Integrating Droplet Digital Lamp for Absolute Quantification of Cancer Biomarkers - Beatriz Oliveira; Bruno Veigas; Alexandra R. Fernandes; Hugo Águas; Rodrigo Martins; Elvira Fortunato; Pedro Viana Baptista (Sensors, 2020)
Microfluidic (MF) advancements have been leveraged toward the development of state-of-the-art platforms for molecular diagnostics, where isothermal amplification schemes allow for further simplification of DNA detection and quantification protocols. The MF integration with loop-mediated isothermal amplification (LAMP) is today the focus of a new generation of chip-based devices for molecular detection, aiming at fast and automated nucleic acid analysis. Here, we combined MF with droplet digital LAMP (ddLAMP) on an all-in-one device that allows for droplet generation, target amplification, and absolute quantification. This multilayer 3D chip was developed in less than 30 minutes by using a low-cost and extremely adaptable production process that exploits direct laser writing technology in “Shrinky-dinks” polystyrene sheets. ddLAMP and target quantification were performed directly on-chip, showing a high correlation between target concentration and positive droplet score. We validated this integrated chip via the amplification of targets ranging from five to 500,000 copies/reaction. Furthermore, on-chip amplification was performed in a 10 µL volume, attaining a limit of detection of five copies/µL under 60 min. This technology was applied to quantify a cancer biomarker, c-MYC, but it can be further extended to any other disease biomarker.

The fundamental importance of basic science: examples of high-impact discoveries from an international Chemistry Network - Lopes, L. G. F.; Sadler, P.J; Bernardes-Génisson, V.; Moura, J. J. G.; Chauvin, R.; Bernhardt, P. V.; Sousa, E. H. S. (Quim Nova, 2020)

Hand-painted Magic Lantern Slides and the 19th-century Colourmen Winsor & Newton - Vilarigues, Márcia; Otero, Vanessa (A Million Pictures: Magic Lantern Slides in the History of Learning, 2020)

Handling CO2 sorption mechanism in PIL@IL composites - Zanatta, M; Lopes, M.; E. J. Cabrita; Bernardes, CES; Corvo, Mc (Journal of CO2 Utilization, 2020)
The mitigation of climate change effects requires the use of alternative materials and technologies to control CO2 atmospheric levels through its capture, storage and use. In this field, the current work presents the evaluation of two poly(ionic liquid)s (PILs) (poly-1-vinyl-3-ethylimidazolium acetate and hydroxide) combined with free ionic liquid (IL) 1-butyl-3-methylimidolium acetate (BMI·OAc) for CO2 capture. The sorption capacity of PIL@IL composites was evaluated under 20 bar of CO2 at 298 K. Nuclear Magnetic Resonance (NMR) spectroscopy allowed quantification of CO2 sorption (physisorption and/or chemisorption) and in situ study of the PIL−CO2 interaction mechanism. NMR in combination with Molecular Dynamics (MD) simulations suggested a 3D organization of PIL composites, maintaining a similar organization to ILs. Also, the use of aqueous solutions of PIL@IL composites was tested, identifying the optimum conditions for water activation (intrinsic water trapped inside IL structure) for chemisorption. As our main contribution, we demonstrate the possibility to control the sorption pathway towards CO2 physisorption, or CO2 conversion (chemisorption) through carbonation (HCO3−/CO32-) according to the PIL/IL ratio, ions structure and water amount. The use of PIL/IL composites is a promising advance for further CO2 reuse approaching a biomimetic carbonation process.
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Human erythrocytes exposure to juglone leads to an increase of superoxide anion production associated with cytochrome b5 reductase uncoupling - Valério, G.N.; Gutierrez-Merino, C.; Nogueira, F.; Moura, I.; Moura, J. J. G.; Samhan-Arias, A. K. (Biochim Biophys Acta Bioenerg, 2020)

Influence of chain length of prepolymers in permanent memory effect of PDLC assessed by solid-state NMR - Mouquinho, Ana; Corvo, Marta C.; Almeida, Pedro L; Feio, Gabriel M.; João Sotomayor (Liquid Crystals, 2020)
The relationship between linear chain (ethylene oxide units) length of polymerisable monomers with morphology, electro-optical properties and 13C nuclear magnetic resonance (NMR) spectroscopy of the corresponding polymer-dispersed liquid crystal (PDLC) films was investigated. The preferred liquid crystal molecule alignment and permanent memory effect of PDLC were greatly influenced by the length of the molecular chain of prepolymers to be incorporated as a polymer matrix. By increasing the number of ethylene oxide in prepolymer chain and maintaining the number of functionalities (polymerisable groups in each monomer molecule), the permanent memory effect of PDLC increased, as proved by solid-state 13C NMR spectroscopy.
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Light trapping in solar cells: simple design rules to maximize absorption - Kezheng Li; Haque, Sirazul; Augusto Martins; Elvira Fortunato; Rodrigo Martins; Mendes, Manuel J.; Christian S. Schuster (Optica, 2020)

Magnetic Precipitation: A New Platform for Protein Purification - Santos, R. dos; Iria, I.; Manuel, A. M.; Leandro, A.P.; Madeira, C.A.C.; Gonçalves, J.; Carvalho, A. L.; Roque, A. C. A. (Biotechnology Journal, 2020)

Microfluidics in Gas Sensing and Artificial Olfaction - Rebordão, G.; S. I. C. J. Palma; Roque, A. C. A. (Sensors, 2020)
Rapid, real-time, and non-invasive identification of volatile organic compounds (VOCs) and gases is an increasingly relevant field, with applications in areas such as healthcare, agriculture, or industry. Ideal characteristics of VOC and gas sensing devices used for artificial olfaction include portability and affordability, low power consumption, fast response, high selectivity, and sensitivity. Microfluidics meets all these requirements and allows for in situ operation and small sample amounts, providing many advantages compared to conventional methods using sophisticated apparatus such as gas chromatography and mass spectrometry. This review covers the work accomplished so far regarding microfluidic devices for gas sensing and artificial olfaction. Systems utilizing electrical and optical transduction, as well as several system designs engineered throughout the years are summarized, and future perspectives in the field are discussed.

Microwave synthesis of silver sulfide and silver nanoparticles: light and time influence - David Magalhães Sousa; William Chiappim; Joaquim P. Leitão; João Carlos Lima; Ferreira, Isabel (ACS omega, 2020)

Natural multimerization rules the performance of affinity-based physical hydrogels for stem cell encapsulation and differentiation - Fernandes, C.S.M.; Rodrigues, A.L.; Alves, V. D.; Fernandes, T. G.; Pina, A. S.; Roque, A. C. A. (Biomacromolecules, 2020)

The Nature of Carbon Dioxide in Bare Ionic Liquid - Zanatta, Marcileia; Nathália M. Simon; Dupont, Jairton (ChemSusChem, 2020)
Ionic liquids (ILs) are among the most studied and promising materials for selective CO2 capture and transformation. The high CO2 sorption capacity associated with the possibility to activate this rather stable molecule through stabilization of ionic/radical species or covalent interactions either with the cation or anion has opened new avenues for CO2 functionalization. However, recent reports have demonstrated that another simpler and plausible pathway is also involved in the sorption/activation of CO2 by ILs associated with basic anions. Bare ILs or IL solutions contain almost invariable significant amounts of water and through interaction with CO2 generate carbonates/bicarbonates rather than carbamic acids or amidates. In these cases, the IL acts as a base and not a nucleophile and yields buffer‐like solutions that can be used to shift the equilibrium toward acid products in different CO2 reutilization reactions. In this Minireview, the emergence of IL buffer‐like solutions as a new reactivity paradigm in CO2 capture and activation is described and analyzed critically, mainly through the evaluation of NMR data.

Nitrate-nitrite fate and oxygen sensing in dormant Mycobacterium tuberculosis: A bioinorganic approach highlighting the importance of transition metals - Sousa, E. H. S.; Carepo, M. S.; Moura, J. J. G. (Coord Chem Rev, 2020)

Noble‐Metal‐Free Memristive Devices Based on IGZO for Neuromorphic Applications - Pereira, Maria; Deuermeier, Jonas; Nogueira, Ricardo; Patricia Almeida Carvalho; Rodrigo Martins; Elvira Fortunato; Kiazadeh, Asal (Advanced electronic Materials, 2020)

Peptide and Protein Engineering: From Concepts to Biotechnological Applications - Iranzo, O.; Roque, A. C. A. (Springer, 2020)

Photonic-structured TCO front contacts yielding optical and electrically enhanced thin-film solar cells - Sánchez-Sobrado Olalla; Mendes, Manuel J.; Mateus, Tiago; João Costa; Nunes, Daniela; Hugo Águas; Elvira Fortunato; Rodrigo Martins (Solar Energy, 2020)

PolySense: Augmenting Textiles with Electrical Functionality using In-Situ Polymerization - C. Honnet; H. Perner-Wilson; M. Teyssier; B. Fruchard; J. Steimle; Baptista, A. C.; P. Strohmeier (Proceedings of the 2020 CHI Conference on Human Factors in Computing Systems, 2020)

PolySense: How to Make Electrically Functional Textiles - P. Strohmeier; C. Honnet; H Pernet-Wilson; M. Teyssier; B. Fruchard; Baptista, A. C.; J. Steimle (CHI Conference on Human Factors in Computing Systems, 2020)

Potential therapeutic approaches for a sleeping pathogen: tuberculosis a case for bioinorganic chemistry - Sousa, E. H. S.; Diógenes, I. C. N.; Lopes, L. G. F.; Moura, J. J. G. (J Biol Inorg Chem, 2020)

Preparation and Characterization of Zinc and Magnesium Doped Bioglasses - Sílvia R. Gavinho; Mariana Castro Soares; Borges, João Borges; Silva, Jorge Carvalho; Isabel Sá Nogueira; Manuel P. Graça (Nanoscience and Nanotechnology in Security and Protection against CBRN Threats, 2020)
Peri-implantitis is an infectious disease that affects about one of five patients who receive a dental implant within 5 years after the surgery. To minimize this reaction the development of new biomaterials with antibacterial action is needed that can be used as a coating material in orthodontic implants. In addition, these biomaterials can be doped with several ions, which add specific properties that may act at the cellular level, such as increasing the angiogenesis efficiency. In this work, 45S5 Bioglass® has been used as the base material because it presents higher bioactivity compared to other biomaterials. To add antibacterial function and increase positive effects on bone metabolism, zinc and magnesium ions were introduced in the glass network. The main objective was the synthesis of the 45S5 glass by melt-quenching and study the biological performance as function of the zinc and magnesium concentrations. The structural and biological properties of the prepared samples are discussed.

Rational design of affinity ligands for bioseparation - Matos, MJB; Pina, A. S.; Roque, A. C. A. (Journal of Chromatography A, 2020)

Rational Use of High-Strength Concrete in Flat Slab-Column Connections under Seismic Loading - Micael Inácio; Brisid Isufi; Massimo Lapi; António Pinho Ramos (ACI Structural Journal, 2020)
High Strength Concrete (HSC) slab–column connections with relatively low concrete strengths compared to today’s capabilities have been tested under seismic-type loading in the past. Herein, the hybrid use of HSC with compressive strength around 120 MPa and Normal Strength Concrete (NSC) is investigated through three reversed horizontal cyclic loading tests with different geometries of the HSC region and a reference NSC specimen. The results show that HSC applied in the vicinity of the column can significantly enhance the seismic performance of slab–column connections. The best result in terms of drift capacity and economic use of HSC was achieved in the case of full-depth HSC extended from the column’s face up to 2.5 times the effective depth. Drift ratios up to 3.0% were achieved. A comparison with previous tests showed that the hybrid use of HSC and NSC can achieve similar results to the provision of punching shear reinforcement.

Reliability and NDT Methods - Santos, T.G.; Oliveira, J.P.; Machado, M.A. (Advanced Structured Materials, 2020)
Composites are finding increased use in structural high demanding and high added value applications in advanced industries. A wide diversity exists in terms of matrix type, which can be either polymeric or metallic and type of reinforcements (ceramic, polymeric or metallic). Several technologies have been used to produce these composites; among them, additive manufacturing (AM) is currently being applied. In structural applications, the presence of defects due to fabrication is of major concern, since it affects the performance of a component with negative impact, which can affect, ultimately, human lives. Thus, the detection of defects is highly important, not only surface defects but also barely visible defects. This chapter describes the main types of defects expected in composites produced by AM. The fundamentals of different non-destructive testing (NDT) techniques are briefly discussed, as well as the state of the art of numerical simulation for several NDT techniques. A multiparametric and customized inspection system was developed based on the combination of innovative techniques in modelling and testing. Experimental validation with eddy currents, ultrasounds, X-ray and thermography is presented and analysed, as well as integration of distinctive techniques and 3D scanning characterization.
https://doi.org/10.1007/978-3-030-44522-5_8

Sauropode (2020)

Seeing the Unseen: The Role of Liquid Crystals in Gas‐Sensing Technologies - Esteves, C; Ramou, E.; Porteira, A.R.P.; Barbosa, AJM; Roque, A. C. A. (Advanced Optical Materials, 2020)
Fast, real-time detection of gases and volatile organic compounds (VOCs) is an emerging research field relevant to most aspects of modern society, from households to health facilities, industrial units, and military environments. Sensor features such as high sensitivity, selectivity, fast response, and low energy consumption are essential. Liquid crystal (LC)-based sensors fulfill these requirements due to their chemical diversity, inherent self-assembly potential, and reversible molecular order, resulting in tunable stimuliresponsive soft materials. Sensing platforms utilizing thermotropic uniaxial systems—nematic and smectic—that exploit not only interfacial phenomena, but also changes in the LC bulk, are demonstrated. Special focus is given to the different interaction mechanisms and tuned selectivity toward gas and VOC analytes. Furthermore, the different experimental methods used to transduce the presence of chemical analytes into macroscopic signals are discussed and detailed examples are provided. Future perspectives and trends in the field, in particular the opportunities for LC-based advanced materials in artificial olfaction, are also discussed.

Self-Cleaned Photonic-Enhanced Solar Cells with Nanostructured Parylene-C - Pedro Centeno; Alexandre, Miguel; Chapa, Manuel; J. V. Pinto; Deuermeier, Jonas; Mateus, Tiago; Elvira Fortunato; Rodrigo Martins; Hugo Águas; Mendes, Manuel J. (Advanced Materials Interfaces, 2020)

Spontaneous Aminolytic Cyclization and Self-Assembly ofDipeptide Methyl Esters in Water - Pappas, C. G.; Wijerathne, N.; Sahoo, J. K.; Jain, A.; Kroiss, D.; Sasselli, I. R.; Pina, A. S.; Lampel, A.; R. V. Ulijn (ChemSystemsChem, 2020)

Sustainable plant polyesters as substrates for optical gas sensors - Rodrigues, R; S. I. C. J. Palma; Correia, V.J.; Padrao, I.; Pais, J.; Banza, M.; Alves, C.; Deuermeier, J.; Martins, C.; Costa, HMA; Ramou, E.; Silva Pereira, C.; et al. (Materials Today Bio, 2020)
The fast and non-invasive detection of odors and volatile organic compounds (VOCs) by gas sensors and electronic noses is a growing field of interest, mostly due to a large scope of potential applications. Additional drivers for the expansion of the field include the development of alternative and sustainable sensing materials. The discovery that isolated cross-linked polymeric structures of suberin spontaneously self-assemble as a film inspired us to develop new sensing composite materials consisting of suberin and a liquid crystal (LC). Due to their stimuliresponsive and optically active nature, liquid crystals are interesting probes in gas sensing. Herein, we report the isolation and the chemical characterization of two suberin types (from cork and from potato peels) resorting to analyses of gas chromatography–mass spectrometry (GC-MS), solution nuclear magnetic resonance (NMR), and Xray photoelectron spectroscopy (XPS). The collected data highlighted their compositional and structural differences. Cork suberin showed a higher proportion of longer aliphatic constituents and is more esterified than potato suberin. Accordingly, when casted it formed films with larger surface irregularities and a higher C/O ratio. When either type of suberin was combined with the liquid crystal 5CB, the ensuing hybrid materials showed distinctive morphological and sensing properties towards a set of 12 VOCs (comprising heptane, hexane, chloroform, toluene, dichlormethane, diethylether, ethyl acetate, acetonitrile, acetone, ethanol, methanol, and acetic acid). The optical responses generated by the materials are reversible and reproducible, showing stability for 3 weeks. The individual VOC-sensing responses of the two hybrid materials are discussed taking as basis the chemistry of each suberin type. A support vector machines (SVM) algorithm based on the features of the optical responses was implemented to assess the VOC identification ability of the materials, revealing that the two distinct suberin-based sensors complement each other, since they selectively identify distinct VOCs or VOC groups. It is expected that such new environmentally-friendly gas sensing materials derived from natural diversity can be combined in arrays to enlarge selectivity and sensing capacity.

Synthesis of Cerium Dioxide Nanoparticles in Aqueous Solution at Controlled pH Values - Shlapa, Yu.; Solopan, S.; Timashkov, I.; Belous, A. (1st International Research and Practice Conference “Nanoobjects & Nanostructuring” N&N-2020, 2020)

Ta2O5/SiO2 Multicomponent Dielectrics for Amorphous Oxide TFTs - https://www.mdpi.com/2673-3978/2/1/1/htm (Electronic Materials, 2020)

Tensile behaviour characterization of a high performance fiber reinforced concrete - João Peixoto (FEUP - Faculdade de Engenharia da Universidade do Porto, 2020)

Teste (2020)

Toward the Mechanistic Understanding of Enzymatic CO2 Reduction - Ana Rita Oliveira; Mota, Cristiano; Mourato, Cláudia; Domingos, Renato M.; Santos, Marino F. A.; Gesto, Diana; Guigliarelli, Bruno; Santos-Silva, Teresa; Romão, Maria João; Pereira, Inês A. Cardoso (ACS Catalysis, 2020)

Toxicity Evaluation of Quantum Dots (ZnS and CdS) Singly and Combined in Zebrafish (Danio rerio) - Beatriz Matos; Marta Martins; Antonio Cid Samamed; David Sousa; Ferreira, Isabel; Mário S. Diniz (International journal of environmental research and public health, 2020)

Unravelling the Secrets of Magic Lantern Slide Painting - Ângela Santos; Otero, Vanessa; Beatriz Rodrigues; Vilarigues, Márcia (2020)

Versatile and Tunable Poly(Ethylene Glycol)‐Based Hydrogels Crosslinked through the Ugi Reaction - Padrão, I.; Fernandes, C.S.M.; Esteves, C; Fernandes, T.; Pina, A. S.; Roque, A. C. A. (ChemPlusChem, 2020)

Wave-optical front structures on silicon and perovskite thin-film solar cells - Mendes, Manuel J.; Sanchez-Sobrado, Olalla; Haque, Sirazul; Mateus, Tiago; Hugo Águas; Elvira Fortunato; Rodrigo Martins (Solar Cells and Light Management: Materials, Strategies and Sustainability, 2020)

Acidic porous carbons involved in the green and selective synthesis of benzodiazepines - M. Godino-Ojer; Matos, I.; M. Bernardo; Carvalho, R.; Olívia Salomé G.P. Soares; C. Durán-Valle; I. M. Fonseca; E. Pérez Mayoral (Catalysis Today, 2020)
Eco-sustainable and recyclable porous carbons are reported as metal-free catalysts for the synthesis of benzodiazepines for the first time. The porous carbons were able to efficiently catalyse the synthesis of benzodiazepine 1 from o-phenylendiamine 2 and acetone 3 under mild conditions. Both acidic functions and the porosity of the catalysts were determinant features. High conversion values were obtained when using HNO3 oxidized carbons. The highest selectivity to benzodiazepine 1 was obtained in the presence of the most microporous catalyst N-N, which is indicative of the great influence of porous properties. Stronger acid sites and high microporosity of the carbon treated with H2SO4 yield benzodiazepine 1 with total selectivity.
SI: CarboCat–VIII
Keywords: activated carbons; benzodiazepines; Heterocyclic chemistry; Heterogeneous catalytic systems

Activation of co-pyrolysis chars from rice wastes to improve the removal of Cr3+ from simulated and real industrial wastewaters - Diogo Dias; Bernardo, Maria; Matos, Inês; Fonseca, Isabel; Filomena Pinto; Nuno Lapa (Journal of Cleaner Production, 2020)
Chromium is one of the most important raw materials for the European Union. Adsorption has become an important process for the recovery of metals from wastewaters, which has led to a demand for low-cost and eco-friendly adsorbents. The objective of this work was to use new and renewable carbon-based adsorbents from rice wastes in the removal/recovery of Cr(III) from synthetic and real wastewaters. Rice wastes were submitted to co-pyrolysis and the resulting char was optimized through physical and/or chemical activations/treatments. A commercial activated carbon was used for comparison purposes. All adsorbents were characterized (including an ecotoxicity test for the char precursor) and submitted to Cr(III) removal assays from a synthetic solution, in which two solid/liquid ratios (S/L) were tested (5 and 10 g/L). The CO2 activated carbon at a S/L = 5 g/L was the biomass-derived adsorbent that performed better, obtaining a maximum Cr(III) uptake capacity of 9.23 mg/g comparable to the one obtained by the commercial adsorbent at the same S/L (9.80 mg/g). The good results on this biomass-derived carbon were due to the effective volatile matter removal during the activation (from 22.7 to 4.25% w/w), which increased both surface area (from <5.0 to 325 m2/g) and ash content (from 30.0 to 40.4% w/w), allowing an increase in Cr(III) removal due to ion exchange mechanism and porosity development. The best adsorbent, under optimized conditions, was also applied to a chromium rich industrial wastewater. The results obtained in this real case application demonstrated a competition effect due to the presence of other ions.
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Keywords: activated carbons; Activation; Adsorption; Cr(III); Pyrolysis; Rice wastes

Assessment of potato peel and agro-forestry biochars supplementation on in vitro ruminal fermentation - Rodrigues, Ana R.F.; Maia, Margarida R.G.; Cabrita, Ana R.J.; Oliveira, Hugo M.; Bernardo, Maria; Nuno Lapa; Fonseca, Isabel; Trindade, Henrique; Pereira, José L.; Fonseca, António J.M. (PeerJ, 2020)
Background The awareness of environmental and socio-economic impacts caused by greenhouse gas emissions from the livestock sector leverages the adoption of strategies to counteract it. Feed supplements can play an important role in the reduction of the main greenhouse gas produced by ruminants—methane (CH4). In this context, this study aims to assess the effect of two biochar sources and inclusion levels on rumen fermentation parameters in vitro. Methods Two sources of biochar (agro-forestry residues, AFB, and potato peel, PPB) were added at two levels (5 and 10%, dry matter (DM) basis) to two basal substrates (haylage and corn silage) and incubated 24-h with rumen inocula to assess the effects on CH4 production and main rumen fermentation parameters in vitro. Results AFB and PPB were obtained at different carbonization conditions resulting in different apparent surface areas, ash content, pH at the point of zero charge (pHpzc), and elemental analysis. Relative to control (0% biochar), biochar supplementation kept unaffected total gas production and yield (mL and mL/g DM, p = 0.140 and p = 0.240, respectively) and fermentation pH (p = 0.666), increased CH4production and yield (mL and mL/g DM, respectively, p = 0.001) and ammonia-N (NH3-N, p = 0.040), and decreased total volatile fatty acids (VFA) production (p < 0.001) and H2 generated and consumed (p ≤ 0.001). Biochar sources and inclusion levels had no negative effect on most of the fermentation parameters and efficiency. Acetic:propionic acid ratio (p = 0.048) and H2 consumed (p = 0.019) were lower with AFB inclusion when compared to PPB. Biochar inclusion at 10% reduced H2 consumed (p < 0.001) and tended to reduce total gas production (p = 0.055). Total VFA production (p = 0.019), acetic acid proportion (p = 0.011) and H2 generated (p = 0.048) were the lowest with AFB supplemented at 10%, no differences being observed among the other treatments. The basal substrate affected most fermentation parameters independently of biochar source and level used. Discussion Biochar supplementation increased NH3-N content, iso-butyric, iso-valeric and valeric acid proportions, and decreased VFA production suggesting a reduced energy supply for microbial growth, higher proteolysis and deamination of substrate N, and a decrease of NH3-N incorporation into microbial protein. No interaction was found between substrate and biochar source or level on any of the parameters measured. Although AFB and PPB had different textural and compositional characteristics, their effects on the rumen fermentation parameters were similar, the only observed effects being due to AFB included at 10%. Biochar supplementation promoted CH4 production regardless of the source and inclusion level, suggesting that there may be other effects beyond biomass and temperature of production of biochar, highlighting the need to consider other characteristics to better identify the mechanism by which biochar may influence CH4 production.
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Keywords: Agro-forestry biochar; Enteric methane; Potato peel biochar; Rumen fermentation; Supplementation level