Non-invasive and fast diagnostic tools based on volatolomics hold great promise in the control of infectious diseases. However, the tools to identify microbial volatile organic compounds (VOCs) discriminating between human pathogens are still missing. Artificial intelligence is increasingly recognised as an essential tool in health sciences. Machine learning algorithms based in support vector machines and features selection tools were here applied to find sets of microbial VOCs with pathogen-discrimination power. Studies reporting VOCs emitted by human microbial pathogens published between 1977 and 2016 were used as source data. A set of 18 VOCs is sufficient to predict the identity of 11 microbial pathogens with high accuracy (77%), and precision (62–100%). There is one set of VOCs associated with each of the 11 pathogens which can predict the presence of that pathogen in a sample with high accuracy and precision (86–90%). The implemented pathogen classification methodology supports future database updates to include new pathogen-VOC data, which will enrich the classifiers. The sets of VOCs identified potentiate the improvement of the selectivity of non-invasive infection diagnostics using artificial olfaction devices.

In this work 2H NMR spectroscopy and 1H NMR relaxometry and diffusometry were used to characterise water order and dynamics in cellulose acetate/silica asymmetric membranes. Two hydrated membranes were characterised allowing the identification of extra ordering of the water molecules and the presence in each membrane of up to two spectral components with different degrees of order and different T1 values. The mechanism behind this order increase was ascribed to the rapid exchange of the water molecules between the pore walls and its interior. T1 relaxometry dispersions allowed for the identification of the relevant mechanisms of pore-confined water motion, with rotations mediated by translational displacements (RMTD) as the dominant mechanism in the low frequency region. Using the RMTD low cut off frequency along with the in situ directly measured diffusion constant it was possible do determine characteristic lengths of correlated water motion in both membranes studied, which fall in ranges compatible with typical pore dimensions in similar membranes.

The present work addressed the challenges of identifying applicable Non-Destructive Testing (NDT) techniques suitable for inspection and materials characterization techniques for Wire and Arc Additive Manufacturing (WAAM) parts. With the view of transferring WAAM to the industry and qualifying the manufacturing process for applications such as structural components, the quality of the produced parts needs to be assured. Thus, the main objective of this paper is to review the main NDT techniques and assess the capability of detecting WAAM defects, for inspection either in a monitoring, in-process or post-process scenario. Radiography and ultrasonic testing were experimentally tested on reference specimens in order to compare the techniques capabilities. Metallographic, hardness and electrical conductivity analysis were also applied to the same specimens for material characterization. Experimental outcomes prove that typical WAAM defects can be detected by the referred techniques. The electrical conductivity measurement may complement or substitute some destructive methods used in AM processing.

The highly intronic nature of protein coding genes in mammals necessitates a co-transcriptional splicing mechanism as revealed by mNET-seq analysis. Immunoprecipitation of MNase-digested chromatin with antibodies against RNA polymerase II (Pol II) shows that active spliceosomes (both snRNA and proteins) are complexed to Pol II S5P CTD during elongation and co-transcriptional splicing. Notably, elongating Pol II-spliceosome complexes form strong interactions with nascent transcripts, resulting in footprints of approximately 60 nucleotides. Also, splicing intermediates formed by cleavage at the 5′ splice site are associated with nearly all spliced exons. These spliceosome-bound intermediates are frequently ligated to upstream exons, implying a sequential, constitutive, and U12-dependent splicing process. Finally, lack of detectable spliced products connected to the Pol II active site in human HeLa or murine lymphoid cells suggests that splicing does not occur immediately following 3′ splice site synthesis. Our results imply that most mammalian splicing requires exon definition for completion.

The purpose of this work was the development of a scalable and easy-to-use electronic noses (E-noses) system architecture for volatile organic compounds sensing, towards the final goal of using several E-noses acquiring large datasets at the same time. In order to accomplish this, each E-nose system is comprised by a delivery system, a detection system and a data acquisition and control system. In order to increase the scalability, the data is stored in a database common to all E-noses. Furthermore, the system was designed so it would only require five simple steps to setup a new E-nose if needed, since the only parameter that needs to be changed is the ID of the new E-nose. The user interacts with a node using an interface, allowing for the control and visualization of the experiment. At this stage, there are three different E-nose prototypes working with this architecture in a laboratory environment.

A new and never yet reported hetero arylidene-9(10H)-anthrone structure (4) was unexpectedly isolated on reaction of 1,2-dimethyl-3-ethylimidazolium iodide (2) and 9-anthracenecarboxaldehyde (3) under basic conditions. Its structure was unequivocally attributed by X-ray crystallography. No cytotoxicity in human healthy fibroblasts and in two different cancer cell lines was observed indicating its applicability in biological systems. Compound 4 interacts with CT-DNA by intercalation between the adjacent base pairs of DNA with a high binding affinity (Kb = 2.0(± 0.20) x 105 M-1) which is 10x higher than that described for doxorubicin (Kb = 3.2 (±0.23) × 104 M-1). Furthermore, compound 4 quenches the fluorescence emission of GelRed-CT-DNA system with a quenching constant (KSV) of 3.3(±0.3) x 103 M-1 calculated by the Stern-Volmer equation.

As consumption of fish and fish-based foods increases, non-destructive monitoring of fish freshness also becomes more prominent. Fish products are very perishable and prone to microbiological growth, not always easily detected by organoleptic evaluation. The analysis of the headspace of fish specimens through gas sensing is an interesting approach to monitor fish freshness. Here we report a gas sensing method for monitoring Tilapia fish spoilage based on the application of a single gas sensitive gel material coupled to an optical electronic nose. The optical signals of the sensor and the extent of bacterial growth were followed over time, and results indicated good correlation between the two determinations, which suggests the potential application of this simple and low cost system for Tilapia fish freshness monitoring.

The aim of this work was to determine the physico-chemical characteristics of the stained-glass windows into the 19th century of two mausoleums located in the city of Belém do Pará (Brazil), and to evaluate their state of conservation. The glass chemical composition was determined by WXRF and SEM/EDS. The samples’ morphology and the microorganisms’ identification were carried out by optical microscopy. The results indicated that the samples were soda-lime silicate glass, with approximately 70wt. % of SiO2, which contributed to the resistance of the stained glass to the weathering. The concentration of Na2O was normally twice the K2O, which contrasts with the composition of other panels produced during the same period, as reported in the literature. The biofilm is composed by cyanobacteria and rotifers. Overall, the panels analysed were in a good state of conservation, despite their exposure to tropical climate conditions for more than a century with no preventive measures whatsoever. Resumen El objetivo del presente trabajo fue determinar las características físico-químicas de las vidrieras del siglo XIX correspondientes a dos mausoleos ubicados en la ciudad de Belém do Pará (Brasil) para evaluar su estado de conservación. La composición química del vidrio fue determinada por WXRF y SEM/EDS. La morfología de las muestras y la identificación de los microorganismos fueron realizadas por microscopia óptica. Los resultados indicaron que las muestras eran vidrios de silicato sódico-cálcico, con aproximadamente el 70% en peso de SiO2, lo que aumentó la resistencia a la corrosión de los vidrios de estas vidrieras. La concentración de Na2O fue normalmente el doble que de K2O, lo que contrasta con la composición de otros paneles producidos durante el mismo período, de acuerdo con la literatura. El biofilm presentó cianobacterias y rotíferas. En general, los paneles analizados presentaban un buen estado de conservación, a pesar de su exposición a las condiciones climáticas tropicales durante más de un siglo, sin las medidas de conservación preventivas.

G. metallireducens bacterium has highly versatile respiratory pathways that provide the microorganism an enormous potential for many biotechnological applications. However, little is known about the structural and functional properties of its electron transfer components. In this work, the periplasmic cytochrome PpcA from G. metallireducens was studied in detail for the first time using complementary biophysical techniques, including UV–visible, CD and NMR spectroscopy. The results obtained showed that PpcA contains three low-spin c-type heme groups with His-His axial coordination, a feature also observed for its homologue in G. sulfurreducens. However, despite the high sequence homology between the two cytochromes, important structural and functional differences were observed. The comparative analysis of the backbone, side chain and heme substituents NMR signals revealed differences in the relative orientation of the hemes I and III. In addition, redox titrations followed by visible spectroscopy showed that the redox potential values for PpcA from G. metallireducens (−78 and −93 mV at pH 7 and 8, respectively) are considerably less negative. Overall, this study provides biochemical and biophysical data of a key cytochrome from G. metallireducens, paving the way to understand the extracellular electron transfer mechanisms in these bacteria.

Highly porous monolithic alumina columns find a wide variety of applications, including in chromatography, due to increased surface area and good accessibility to the ligands and reduced diffusional hindrances. Several modelling approaches have been applied to describe experimentally observed flow behaviour in such materials, which morphology plays a key role in determining their hydrodynamic and mass transfer properties. In this work, a direct computational fluid dynamics (CFD) modelling approach is proposed to simulate flow behaviour in monolithic porous columns. The morphological structure of a fabricated alumina monolith was first reconstructed using 3D X-ray tomography data and, subsequently, OpenFOAM, an open-source CFD tool, was used to simulate the essential parameters for monoliths’ performance characterisation and optimisation, i.e. velocity and pressure fields, fluid streamlines, shear stress and residence time distribution (RTD). Moreover, the tortuosity of the monolith was estimated by a novel method, using the computed streamlines, and its value (∼1.1) was found to be in the same range of the results obtained by known experimental, analytical and numerical equations. Besides, it was observed (for the case of the monolith studied) that fluid transport was dominated by flow heterogeneities and advection, while the shear stress at pore mouths was significantly higher than in other regions. The proposed modelling approach, with expected high potential for designing target materials, was successfully validated by an experimentally obtained residence time distribution (RTD).

The Cathedral of León has one of the most important ensembles of medieval stained glass windows in Europe; however, most of them have been altered by atmospheric weathering. The main objective of this study was the characterization of a set of glass samples from the Cathedral of León, the comparison with glasses from previous interventions in the cathedral and the study of the relation between the alteration pathologies, the chemical composition of the glasses and the environment in which they were placed. The samples were characterized by means of binocular microscopy, scanning electron microscopy and energy dispersive X-ray spectrometry, visible spectrophotometry, X-ray fluorescence spectrometry and X-ray diffraction. The main alteration of glasses exposed until the 19th century was the formation of pits by dealkalinization, while the glasses exposed until the present formed CaSO4 deposits as a consequence of the synergic effect of rainwater and gaseous pollutants. Glasses altered by the browning of manganese were also characterized. Resumen La Catedral de León posee una de las colecciones de vidrieras medievales más importantes de Europa; sin embargo, muchos de los vidrios se han alterado por degradación atmosférica. El objetivo principal de este estudio fue la caracterización de un conjunto de vidrios procedentes de la Catedral de León, su comparación con vidrios procedentes de intervenciones previas en la catedral y el estudio de la relación entre las patologías de alteración, la composición química del vidrio y el medio en el que estuvieron expuestas. Las muestras se caracterizaron mediante lupa binocular, microscopía electrónica de barrido y microanálisis de dispersión de energía de rayos X, espectrofotometría visible, espectrometría de fluorescencia de rayos X y difracción de rayos X. La alteración principal de los vidrios expuestos hasta el s. xix consistió en la formación de picaduras por desalcalinización, mientras que en los vidrios expuestos hasta la actualidad se formaron depósitos de CaSO4 como consecuencia del efecto sinérgico de la lluvia y los gases contaminantes. También se han caracterizado vidrios alterados por el enmarronamiento del manganeso.

Blends of rice waste streams were submitted to co-gasification assays. The resulting chars (G1C and G2C) were characterized and used in Cr(III) removal assays from a synthetic solution. A Commercial Activated Carbon (CAC) was used for comparison purposes. The chars were non-porous materials mainly composed by ashes (68.3–92.6% w/w). The influences of adsorbent loading (solid/liquid ratio – S/L) and initial pH in Cr(III) removal were tested. G2C at a S/L of 5 mg L−1 and an initial pH of 4.50 presented an uptake capacity significantly higher than CAC (7.29 and 2.59 mg g−1, respectively). G2C was used in Cr(III) removal assays from an industrial wastewater with Cr(III) concentrations of 50, 100 and 200 mg L−1. Cr(III) removal by precipitation (uptake capacity ranging from 11.1 to 14.9 mg g−1) was more effective in G2C, while adsorption (uptake capacity of 16.1 mg g−1) was the main removal mechanism in CAC.

This paper studied the alteration of three soda-rich-silicate glass types (Cristallo, façon-de-Venise and soda-lime silicate). Replica samples were exposed to different environments simulating museum-like conditions – room temperature and different relative humidities (RH). Results were analysed by SIMS, μ-FTIR, Optical Microscopy and Optical 3D profilometry. Cristallo appears as the most vulnerable, whilst the soda-lime glass appears as the most resistant to the environmental deterioration. The thickness of the altered layer is proportional to the time of exposure and to the RH of the surrounding atmosphere. From the results obtained, the glass composition and the water available and adsorbed to the surface strongly influence the kinetics of the surface alteration.

Abstract This work reports for the first time a new series of promising porous catalytic carbon materials, functionalized with Lewis and Brønsted acid sites useful in the green synthesis of 2,3-dihydro-1H-1,5-benzodiazepine – nitrogen heterocyclic compounds. Benzodiazepines and derivatives are fine chemicals exhibiting interesting therapeutic properties. Carbon materials have been barely investigated in the synthesis of this type of compounds. Two commercial carbon materials were selected exhibiting different textural properties: i) Norit RX3 (N) as microporous sample and ii) mesoporous xerogel (X), both used as supports of ZrO2 (Zr) and ZrO2/SO42− (SZr). The supported SZr led to higher conversion values and selectivities to the target benzodiazepine. Both chemical and textural properties influenced significantly the catalytic performance. Particularly relevant are the results concerning the non-sulfated samples, NZr and XZr, that were able to catalyze the reaction leading to the target benzodiazepine with high selectivity (up to 80 %; 2 h). These results indicated an important role of the carbon own surface functional groups, avoiding the use of H2SO4. Even very low amounts of SZr supported on carbon reveal high activity and selectivity. Therefore, the carbon materials herein reported can be considered an efficient and sustainable alternative bifunctional catalysts for the benzodiazepine synthesis.

This work presents the results of the exposure of soda-lime, potash-lime and mixed-alkali silicate glasses during ten and twenty months in different Portuguese monuments with historical stained-glass windows to characterize the influence of local environmental conditions. The glass samples were exposed in the Monastery of Batalha (Batalha), the Monastery of Jerónimos (Lisbon), and the Cathedral of Évora (Évora). A set of analytical techniques to assess the physicochemical effects were used, including optical microscopy and Fourier transform infrared spectroscopy. All the samples presented crystalline deposits on their surface; however, their quantity and nature depended on the atmospheric conditions during the days before the collection. Potash-lime silicate glass was the most altered glass in comparison with soda-lime and mixed-alkali silicate glasses. The samples from the Cathedral of Évora showed a high content of dust and salts on their surface but without severe chemical pathologies; however, those samples exposed in the Monastery of Jerónimos and the Monastery of Batalha presented alteration layers due to a high humidity environment.

The bacterium Geobacter sulfurreducens can transfer electrons to quinone moieties of humic substances or to anthraquinone-2,6-disulfonate (AQDS), a model for the humic acids. The reduced form of AQDS (AH2QDS) can also be used as energy source by G. sulfurreducens. Such bidirectional utilization of humic substances confers competitive advantages to these bacteria in Fe(III) enriched environments. Previous studies have shown that the triheme cytochrome PpcA from G. sulfurreducens has a bifunctional behavior toward the humic substance analogue. It can reduce AQDS but the protein can also be reduced by AH2QDS. Using stopped-flow kinetic measurements we were able to demonstrate that other periplasmic members of the PpcA-family in G. sulfurreducens (PpcB, PpcD and PpcE) also showed the same behavior. The extent of the electron transfer is thermodynamically controlled favoring the reduction of the cytochromes. NMR spectra recorded for 13C,15N-enriched samples in the presence increasing amounts of AQDS showed perturbations in the chemical shift signals of the cytochromes. The chemical shift perturbations on cytochromes backbone NH and 1H heme methyl signals were used to map their interaction regions with AQDS, showing that each protein forms a low-affinity binding complex through well-defined positive surface regions in the vicinity of heme IV (PpcB, PpcD and PpcE) and I (PpcE). Docking calculations performed using NMR chemical shift perturbations allowed modeling the interactions between AQDS and each cytochrome at a molecular level. Overall, the results obtained provided important structural-functional relationships to rationalize the microbial respiration of humic substances in G. sulfurreducens.

Abstract The family 81 glycoside hydrolase (GH81) from Clostridium thermocellum is a β-1,3-glucanase belonging to cellulosomal complex. The gene encoding \{GH81\} from Clostridium thermocellum (CtLam81A) was cloned and expressed displaying a molecular mass of  82 kDa. CtLam81A showed maximum activity against laminarin (100 U/mg), followed by curdlan (65 U/mg), at pH 7.0 and 75 °C. CtLam81A displayed Km, 2.1 ± 0.12 mg/ml and Vmax, 109 ± 1.8 U/mg, against laminarin under optimized conditions. CtLam81A activity was significantly enhanced by Ca2+ or Mg2+ ions. Melting curve analysis of CtLam81A showed an increase in melting temperature from 91 °C to 96 °C by Ca2+ or Mg2+ ions and decreased to 82 °C by EDTA, indicating that Ca2+ and Mg2+ ions may be involved in catalysis and in maintaining structural integrity. \{TLC\} and MALDI-TOF analysis of β-1,3-glucan hydrolysed products released initially, showed β-1,3-glucan-oligosaccharides degree of polymerization (DP) from \{DP2\} to DP7, confirming an endo-mode of action. The catalytically inactive mutant CtLam81A-E515A generated by site-directed mutagenesis was co-crystallized and tetragonal crystals diffracting up to 1.4 Å resolution were obtained. CtLam81A-E515A contained 15 α-helices and 38 β-strands forming a four-domain structure viz. a β-sandwich domain I at N-terminal, an α/β-domain II, an (α/α)6 barrel domain III, and a small 5-stranded β-sandwich domain IV.

Salinity gradient energy is currently attracting growing attention among the scientific community as a renewable energy source. In particular, Reverse Electrodialysis (RED) is emerging as one of the most promising membrane-based technologies for renewable energy generation by mixing two solutions of different salinity. This work presents a critical review of the most significant achievements in RED, focusing on membrane development, stack design, fluid dynamics, process optimization, fouling and potential applications. Although RED technology is mainly investigated for energy generation from river water/seawater, the opportunities for the use of concentrated brine are considered as well, driven by benefits in terms of higher power density and mitigation of adverse environmental effects related to brine disposal. Interesting extensions of the applicability of RED for sustainable production of water and hydrogen when complemented by reverse osmosis, membrane distillation, bio-electrochemical systems and water electrolysis technologies are also discussed, along with the possibility to use it as an energy storage device. The main hurdles to market implementation, predominantly related to unavailability of high performance, stable and low-cost membrane materials, are outlined. A techno-economic analysis based on the available literature data is also performed and critical research directions to facilitate commercialization of RED are identified.

The plant cell-wall is constituted by structurally diverse polysaccharides. The biodegradation of these is a crucial process for life sustainability. Cellulolytic microorganisms are highly efficient in this process by assembling modular architectures of carbohydrate-active enzymes with appended non-catalytic carbohydrate-binding modules (CBMs). Carbohydrate microarrays offer high-throughput and sensitive tools for uncovering carbohydrate-binding specificities of CBMs{,} which is pivotal to understand the function of these modules in polysaccharide biodegradation mechanisms. Features of this technology will be here briefly reviewed with highlights of microarray approaches to study plant-carbohydrates and CBM-carbohydrate interactions{,} along with an overview of plant polysaccharides and microorganisms strategies for their recognition.

Here we describe the establishment of a new canine mammary tumour (CMT) cell line, FR37-CMT that does not show dependence on female hormonal signaling to induce tumour xenografts in NOD-SCID mice. FR37-CMT cell line has a stellate or fusiform shape, displays the ability to reorganize the collagen matrix, expresses vimentin, CD44 and shows the loss of E-cadherin which is considered a fundamental event in epithelial to mesenchymal transition (EMT). The up-regulation of ZEB1, the detection of phosphorylated ERK1/2 and the downregulation of DICER1 and miR-200c are also in accordance with the mesenchymal characteristics of FR37-CMT cell line. FR37-CMT shows a higher resistance to cisplatin (IC50>50 µM) and to doxorubicin (IC50>5.3 µM) compared with other CMT cell lines. These results support the use of FR37-CMT as a new CMT model that may assist the understanding of the molecular mechanisms underlying EMT, CMT drug resistance, fostering the development of novel therapies targeting CMT.

Photothermal Therapy (PTT) impact in cancer therapy has been increasing due to the enhanced photothermal capabilities of a new generation of nanoscale photothermal agents. Among these nanoscale agents, gold nanoshells and nanorods have demonstrated optimal properties for translation of near infra-red radiation into heat at the site of interest. However, smaller spherical gold nanoparticles (AuNPs) are easier to produce, less toxic and show improved photoconversion capability that may profit from the irradiation in the visible via standard surgical green lasers. Here we show the efficient light-to-heat conversion of spherical 14 nm AuNPs irradiated in the visible region (at the surface plasmons resonance peak) and its application to selectively obliterate cancer cells. Using breast cancer as model, we show a synergistic interaction between heat (photoconversion at 530 nm) and cytotoxic action by doxorubicin with clear advantages to those of the individual therapy approaches.