In the present work composite nanoparticles with a magnetic core and a chitosan-based shell were produced as drug delivery systems for doxorubicin (DOX). The results show that composite nanoparticles with a hydrodynamic diameter within the nanometric range are able to encapsulate more DOX than polymeric nanoparticles alone corresponding also to a higher drug release. Moreover the synthesis method of the iron oxide nanoparticles influences the total amount of DOX released and a high content of iron oxide nanoparticles inhibits DOX release. The modelling of the experimental results revealed a release mechanism dominated by Fickian diffusion.

The congeneric freshwater fish Squalius carolitertii and S. torgalensis inhabit different Iberian regions with distinct climates; Atlantic in the North and Mediterranean in the South, respectively. While northern regions present mild temperatures, fish in southern regions often experience harsh temperatures and droughts. Previous work with two hsp70 genes suggested that S. torgalensis is better adapted to harsher thermal conditions than S. carolitertii as a result of the different environmental conditions. We present a transcriptomic characterisation of these species' thermal stress responses. Through differential gene expression analysis of the recently available transcriptomes of these two endemic fish species, comprising 12 RNA-seq libraries from three tissues (skeletal muscle, liver and fins) of fish exposed to control (18 °C) and test (30 °C) conditions, we intend to lay the foundations for further studies on the effects of temperature given predicted climate changes. Results showed that S. carolitertii had more upregulated genes, many of which are involved in transcription regulation, whereas S. torgalensis had more downregulated genes, particularly those responsible for cell division and growth. However, both species displayed increased gene expression of many hsps genes, suggesting that they are able to deal with protein damage caused by heat, though with a greater response in S. torgalensis. Together, our results suggest that S. torgalensis may have an energy saving strategy during short periods of high temperatures, re-allocating resources from growth to stress response mechanisms. In contrast, S. carolitertii regulates its metabolism by increasing the expression of genes involved in transcription and promoting the stress response, probably to maintain homoeostasis. Additionally, we indicate a set of potential target genes for further studies that may be particularly suited to monitoring the responses of Cyprinidae to changing temperatures, particularly for species living in similar conditions in the Mediterranean Peninsulas.

A common strategy for the production and purification of recombinant proteins is to fuse a tag to the protein terminal residues and employ a “tag-specific” ligand for fusion protein capture and purification. In this work, we explored the effect of two tryptophan-based tags, NWNWNW and WFWFWF, on the expression and purification of Green Fluorescence Protein (GFP) used as a model fusion protein. The titers obtained with the expression of these fusion proteins in soluble form were 0.11 mg ml−1 and 0.48 mg ml−1 for WFWFWF and NWNWNW, respectively. A combinatorial library comprising 64 ligands based on the Ugi reaction was prepared and screened for binding GFP-tagged and non-tagged proteins. Complementary ligands A2C2 and A3C1 were selected for the effective capture of NWNWNW and WFWFWF tagged proteins, respectively, in soluble forms. These affinity pairs displayed 106 M−1 affinity constants and Qmax values of 19.11 ± 2.60 ug g−1 and 79.39 ug g−1 for the systems WFWFWF AND NWNWNW, respectively. GFP fused to the WFWFWF affinity tag was also produced as inclusion bodies, and a refolding-on column strategy was explored using the ligand A4C8, selected from the combinatorial library of ligands but in presence of denaturant agents.

Improvements of energy efficiency and reduction of Electricity Consumption (EC) could be pushed by increased knowledge on consumption profiles. This paper contributes to a comprehensive understanding of the EC profiles in a Southwest European city through the combination of high-resolution data from smart meters (daily electricity consumption) with door-to-door 110-question surveys for a sample of 265 households in the city of Évora, in Portugal. This analysis allowed to define ten power consumption clusters using Ward's method hierarchical clustering, corresponding to four distinct types of annual consumption profiles: U shape (sharp and soft), W shape and Flat. U shape pattern is the most common one, covering 77% of the sampled households.
The results show that three major groups of determinants characterize the electricity consumption segmentation: physical characteristics of a dwelling, especially year of construction and floor area; HVAC equipment and fireplaces ownership and use; and occupants’ profiles (mainly number and monthly income).
The combination of the daily EC data with qualitative door-to-door survey-based data proved to be a powerful data nutshell to distinguish groups of power consumers, allowing to derive insights to support DSOs, ESCOs, and retailers to design measures and instruments targeted to effective energy reduction (e.g. peak shaving, energy efficiency).

The extracellular electron transfer metabolism of Geobacter sulfurreducens is sustained by several multiheme c-type cytochromes. One of these is the dodecaheme cytochrome GSU1996 that belongs to a new sub-class of c-type cytochromes. GSU1996 is composed by four similar triheme domains (A-D). The C-terminal half of the molecule encompasses the domains C and D, which are connected by a small linker and the N-terminal half of the protein contains two domains (A and B) that form one structural unit. It was proposed that this protein works as an electrically conductive device in Geobacter sulfurreducens, transferring electrons within the periplasm or to outer-membrane cytochromes. In this work, a novel strategy was applied to characterize in detail the thermodynamic and kinetic properties of the hexaheme fragment CD of GSU1996. This characterization revealed the electron transfer process of GSU1996 for the first time, showing that a heme at the edge of the C-terminal of the protein is thermodynamic and kinetically competent to receive electrons from physiological redox partners. This information contributes towards understanding how this new sub-class of cytochromes functions as nanowires, and also increases the current knowledge of the extracellular electron transfer mechanisms in Geobacter sulfurreducens.

This work reports the optimization of V2O5 Seebeck coefficient to obtain high sensitivity and transparent temperature sensors. It is observed that the film thickness plays a major role on the thermoelectric properties, together with the annealing step, obtaining a Seebeck coefficient of −690 μV K−1, for 75 nm thick V2O5 films deposited on glass, after an annealing step of 1 h at 773 K, in air. The V2O5 films are also deposited and optimized on polyimide substrates, but lower annealing temperature is required, 573 K for 3 h, to maintain the flexibility of the substrate and simultaneously high Seebeck coefficient, −591 μV K−1. These films are used in a simple design sensor and tested on the surface of a microfluidic channel (500 μm) made of polydimethylsiloxane, while having hot water flowing through it. The response time is below 1 s and the recovery time around 5 s.

The thermoelectric (TE) properties of vanadium pentoxide (V2O5) alloyed with graphite (G) were studied as a function of its incorporation percentage. Variable weight percentages of graphite powder (0–50%) were added to V2O5 powder and their mixtures were evaporated by a thermal evaporation technique to form thin films with a thickness in the range of 30–80 nm. In the infrared wavelength region, the transmittance of the obtained films increased as the G percentage was increased, while in the visible range, it decreased with G up to 10%. The TE properties were improved when G was in the range of 10–30%, while it decreased for the other percentages: Seebeck coefficient (S) changed from 0.6 mV/K to 0.9 mV/K and was zero with a G of 50%; the electrical conductivity varied slightly from 5 (Ωm)−1 to 0.7 (Ωm)−1 while the mobility improved from 0.07 cm2/V s to 1.5 cm2/V s and the respective carrier concentration was reduced, from 1 × 1018 cm−3 to 4 × 1016 cm−3. These films were applied as temperature sensors evaluating the thermovoltage as a function of thermal gradient between two electrodes, in which one was maintained at room temperature.

Two water soluble heterometallic potassium–dioxidovanadium polymers, [KVO2(L1)]n (1) and [KVO2(L2)(H2O)]n (2) [H2L1= (2,3-dihydroxybenzylidene)-2-hydroxybenzohydrazide and H2L2=(2,3-dihydroxybenzylidene)benzohydrazide], have been synthesized and characterized by IR, NMR, elemental analysis and single crystal X-ray diffraction. The antiproliferative potentials of 1 and 2 were examined towards human colorectal carcinoma (HCT116), and lung (A549) and breast (MCF7) adenocarcinoma cell lines. 1 exhibits a high cytotoxic activity against colorectal carcinoma cells (HCT116), with IC50 lower than those for cisplatin.

The pharmaceutical drug naproxen was loaded in three different silica hosts with pore diameters of 2.4 (MCM), 3.2 (MCM), and 5.9 nm (SBA), respectively: napMCM\_2.4 nm, napMCM\_3.2 nm, and napSBA\_5.9 nm. To access the guest physical state in the prepared composites, differential scanning calorimetry (DSC), X-ray diffraction (XRD), and attenuated total reflectance Fourier transform infrared spectroscopy were used. The different techniques provided complementary information on a molecular population that was revealed to be distributed among different environments, namely the pore core, the inner pore wall, and the outer surface. It was found that naproxen is semicrystalline in the higher pore size matrix being able to crystallize inside pores; after melting it undergoes full amorphization. In the case of the lower pore size matrix, naproxen crystallizes outside pores due to an excess of filling while most of the remaining fraction is incorporated inside the pores as amorphous. Crystallinity in these two composites was observed by the emergence of the Bragg peaks in the XRD analysis, whereas for napMCM\_3.2 nm only the amorphous halo was detected. The latter only exhibits the step due to the glass transition by DSC remaining stable as amorphous at least for 12 months. The glass transition in the three composites is abnormally broad, shifting to higher temperatures as the pore size decreases, coherent with the slowing down of molecular mobility as probed by dielectric relaxation spectroscopy. For napSBA\_5.9 nm the dielectric response was deconvoluted in two processes: a hindered surface (S-) process due to molecules interacting with the inner pore wall and a faster α-relaxation associated with the dynamic glass transition due to molecules relaxing in the pore core, which seems a manifestation of true confinement effects. The drug incorporation inside a nanoporous matrix, mainly in 3.2 nm pores, was revealed to be a suitable strategy to stabilize the highly crystallizable drug naproxen in the amorphous/supercooled state and to control its release from the silica matrix, allowing full delivery after 90 min in basic media.

The WW domain derived from human Yes-associated protein (hYAP65_WW) recognizes proline-rich peptides. The structural and chemical robustness of WW domains makes them appealing candidates to target and capture these peptides in affinity purification processes. In this work{,} the chemical synthesis of the hYAP65_WW domain containing a terminal cysteine for oriented coupling onto the chromatographic matrix was successfully achieved by a fragment solution condensation reaction and by incorporation of pseudoproline dipeptide units. Both strategies yielded a hYAP65_WW protein with the characteristic WW domain folding. The purified hYAP65_WW domain was immobilized in a chromatographic matrix and tested for binding to a proline-rich peptide. The adsorbent bound 92 ng of peptide per mg of support and the elution was particularly efficient when employing a low pH or an increase in salt concentration. This work sets the ground for the application of WW domains as affinity reagents towards the capture and elution of peptides and proteins rich in proline sequences.

Biodiesel production generates low particle size rapeseed waste (recovered from warehouse air filtration systems) that was herein explored as promising biomass precursor of chemically activated carbons. The influence of several experimental parameters on the porosity development was investigated. No benefit was observed when solution impregnation was made nor a significant dependence of the biomass : K2CO3 ratio was observed and{,} as expected{,} high porosity development was obtained only for treatments at 700 [degree]C. Microporous materials with apparent surface area around 1000 m2 g-1 were obtained comparing favorably with literature data regarding activated carbons from rapeseed processing by-products. A selected lab-made sample and two commercial carbons were tested as adsorbents of caffeine from aqueous solution. Although commercial materials present a quicker adsorption rate{,} regarding adsorption capacity the lab-made sample reaches the same value attained by a benchmark material. The regeneration tests made over the rapeseed derived carbon through heat treatments at 600 [degree]C for 1 hour under N2 flow proved that at least two exhaustion-regeneration cycles can be made since the material retains a caffeine adsorption capacity similar to that of the fresh carbon. Therefore{,} a waste management problem of biodiesel industry - rapeseed residue - can be transformed in a valuable material with promising properties for environmental remediation processes.

The conversion of cellulose into products with higher added value often includes a depolymerization step to obtain glucose, its fundamental unity. The depolymerization reaction is carried out via hydrolysis of the β-1,4-glycosidic bond. The search for a solid acid catalyst capable of breaking these bonds is gaining increasing prominence in the literature. In this regard, sulfonated carbons have shown promising results. This work evaluated the use of a residue from the extraction of palm oil as raw material for the production of sulfonated carbons. The raw material was carbonized and sulfonated. The obtained solid acids were tested in the hydrolysis of cellobiose, a dimer of glucose often used as a model compound for cellulose. The hydrolysis reaction is the first step in converting renewable carbon sources into chemical products and biofuels. Some aspects were investigated, as the effect of carbonization temperature on the concentration of sulfonic groups, the results showing that the content thereof reached a maximum value at 300°C. Regarding the hydrolysis of cellobiose, it has been identified that there is a relationship between the concentration of sulfonic acid groups and the activity of these catalysts. However, there is a drop in the turnover number as the amount of sulfonic acid sites increases. This was related to a preferred position sulfonation mechanism. Furthermore, sulfonated carbons showed higher activity than the commercial acid resins, indicating that this material may be a good option for the generation of solid acid catalysts.

Clofibric acid is the metabolite and active principle of blood lipid regulators, it represents the class of acidic pharmaceuticals, and is one of the most persistent drug residues detected in the aquatic environment worldwide. This interdisciplinary work evaluates the effect of solution pH and water hardness in clofibric acid adsorption onto commercial activated carbons. Kinetic and equilibrium assays revealed that the highest clofibric acid removal efficiencies (>70%) were attained at pH 3, and that at pH 8 water hardness degree plays a fundamental role in the adsorption process. In hard water at pH 8 the removal efficiency values increased by 22 or 46% points depending on the carbon sample. Adsorbents’ textural properties also affect the adsorption process since for the microporous sample (CP) the increase of water hardness has a great influence in kinetic and equilibrium data, while for the micro+mesoporous carbon (VP) the variation of the water hardness promoted less significant changes. At pH 3 the increase of water hardness leads to changes in the adsorption mechanism of clofibric acid onto CP carbon signaled by a transition from an S-type to an L-type curve. At pH 8 the change from deionized water to hard water doubles the maximum adsorption capacity of sample CP (101.7mgg−1 vs 211.9mgg−1, respectively). The adsorption enhancement, with water hardness under alkaline conditions, was reasoned in terms of calcium complexation with clofibrate anion exposed by molecular modeling and conductivity studies. Ca2+ complexation by other acidic organic compounds may also occur, and should be considered, since it can play a fundamental role in improved design of water treatment processes employing activated carbons.

Identification of novel molecules that can selectively inhibit the growth of tumor cells, avoid causing side effects to patients and/or intrinsic or acquired resistance, usually associated with common chemotherapeutic agents, is of utmost importance. Organometallic compounds have gained importance in oncologic chemotherapy, such as organotin(IV) complexes. In this study, we assessed the anti-tumor activity of the cyclic trinuclear organotin(IV) complex with an aromatic oximehydroxamic acid group [nBu(2)Sn(L)](3)(H2L = N,2-dihydroxy-5-[N-hydroxyethanimidoyl]benzamide) - MG85 - and provided further characterization of its biological targets. We have previously shown the high anti-proliferative activity of this complex against human colorectal and hepatocellular carcinoma cell lines and lower cytotoxicity in neonatal non-tumor fibroblasts. MG85 induces tumor cell apoptosis and down-regulation of proteins related to tubulin dynamics (TCTP and COF1). Further characterization included the: (i) evaluation of interference in the cell cycle progression, including the expression of critical genes; (ii) affinity to DNA and the corresponding mode of binding; (iii) genotoxic potential in cells with deficient DNA repair pathways; and (iv) in vivo tumor reduction efficiency using mouse colorectal carcinoma xenografts.

The kinetic parameters of the adsorption process at ``liquid–solid'' interface have been evaluated through the sets of time-based experiments of the Cr(III) adsorption under varying temperature, initial metal concentration, and carbon loading for two sets of the commercially available activated carbons and their post-oxidized forms with different texture and surface functionality.

Aldehyde oxidase (AOX1) is an enzyme with broad substrate specificity, catalyzing the oxidation of a wide range of endogenous and exogenous aldehydes as well as N-heterocyclic aromatic compounds. In humans, the enzyme’s role in phase I drug metabolism has been established and its importance is now emerging. However, the true physiologic function of AOX1 in mammals is still unknown. Further, numerous single-nucleotide polymorphisms (SNPs) have been identified in human AOX1. SNPs are a major source of interindividual variability in the human population, and SNP-based amino acid exchanges in AOX1 reportedly modulate the catalytic function of the enzyme in either a positive or negative fashion. For the reliable analysis of the effect of amino acid exchanges in human proteins, the existence of reproducible expression systems for the production of active protein in ample amounts for kinetic, spectroscopic, and crystallographic studies is required. In our study we report an optimized expression system for hAOX1 in Escherichia coli using a codon-optimized construct. The codon-optimization resulted in an up to 15-fold increase of protein production and a simplified purification procedure. The optimized expression system was used to study three SNPs that result in amino acid changes C44W, G1269R, and S1271L. In addition, the crystal structure of the S1271L SNP was solved. We demonstrate that the recombinant enzyme can be used for future studies to exploit the role of AOX in drug metabolism, and for the identification and synthesis of new drugs targeting AOX when combined with crystallographic and modeling studies.

In this work, peptides designed to selectively interact with cellular receptors involved in the regulation of angiogenesis were anchored to oligo-ethylene glycol-capped gold nanoparticles (AuNPs) and used to evaluate the modulation of vascular development using an ex ovo chick chorioallantoic membrane assay. These nanoparticles alter the balance between naturally secreted pro- and antiangiogenic factors, under various biological conditions, without causing toxicity. Exposure of chorioallantoic membranes to AuNP-peptide activators of angiogenesis accelerated the formation of new arterioles when compared to scrambled peptide-coated nanoparticles. On the other hand, antiangiogenic AuNP-peptide conjugates were able to selectively inhibit angiogenesis in vivo. We demonstrated that AuNP vectorization is crucial for enhancing the effect of active peptides. Our data showed for the first time the effective control of activation or inhibition of blood vessel formation in chick embryo via AuNP-based formulations suitable for the selective modulation of angiogenesis, which is of paramount importance in applications where promotion of vascular growth is desirable (eg, wound healing) or ought to be contravened, as in cancer development.

Mammalian aldehyde oxidases (AOXs; EC1.2.3.1) are a group of conserved proteins belonging to the family of molybdo-flavoenzymes along with the structurally related xanthine dehydrogenase enzyme. AOXs are characterized by broad substrate specificity, oxidizing not only aromatic and aliphatic aldehydes into the corresponding carboxylic acids, but also hydroxylating a series of heteroaromatic rings. The number of AOX isoenzymes expressed in different vertebrate species is variable. The two extremes are represented by humans, which express a single enzyme (AOX1) in many organs and mice or rats which are characterized by tissue-specific expression of four isoforms (AOX1, AOX2, AOX3, and AOX4). In vertebrates each AOX isoenzyme is the product of a distinct gene consisting of 35 highly conserved exons. The extant species-specific complement of AOX isoenzymes is the result of a complex evolutionary process consisting of a first phase characterized by a series of asynchronous gene duplications and a second phase where the pseudogenization and gene deletion events prevail. In the last few years remarkable advances in the elucidation of the structural characteristics and the catalytic mechanisms of mammalian AOXs have been made thanks to the successful crystallization of human AOX1 and mouse AOX3. Much less is known about the physiological function and physiological substrates of human AOX1 and other mammalian AOX isoenzymes, although the importance of these proteins in xenobiotic metabolism is fairly well established and their relevance in drug development is increasing. This review article provides an overview and a discussion of the current knowledge on mammalian AOX.

Reactions between 4'-phenyl-terpyridine (L) and several Cu(II) salts (p-toluenesulfonate, benzoate and o-, m-or p-hydroxybenzoate) led to the formation of [Cu(p-SO3C6H4CH3)L(H2O)(2)](p-SO3C6H4CH3) (1), [Cu(OCOPh)(2)L] (2), [Cu(o-OCOC6H4OH)(2)L] (3), [Cu(m-OCOC6H4OH)(2)L]center dot MeOH (4 center dot MeOH) and [Cu(pOCOC(6)H(4)OH)(2)L]center dot 2H(2)O (5 center dot 2H2O), which were characterized by elemental and TG-DTA analyses, ESI-MS, IR spectroscopy and single crystal X-ray diffraction, as well as by conductivimetry. In all structures the Cu atoms present N3O3 octahedral coordination geometries, which, in 2-5, are highly distorted as a result of the chelating-bidentate mode of one of the carboxylate ligands. Intermolecular pi...pi stacking interactions could also be found in 2-5 (in the 3.569-3.651 angstrom range and involving solely the pyridyl rings). Mediumstrong hydrogen bond interactions lead to infinite 1D chains (in 1 and 4) and to an infinite 2D network (in 5). Compounds 1 and 4 show high in vitro cytotoxicity towards HCT116 colorectal carcinoma and HepG2 hepatocellular carcinoma cell lines. The antiproliferative potential of compound 1 is due to an increase of the apoptotic process that was confirmed by Hoechst staining, flow cytometry and RT-qPCR. All compounds able to non-covalently intercalate the DNA helix and induce in vitro pDNA double-strand breaks in the absence of H2O2. Concerning compound 1, the hydroxyl radical and singlet oxygen do not appear to be involved in the pDNA cleavage process and the fact that this cleavage also occurs in the absence of molecular oxygen points to a hydrolytic mechanism of cleavage.

The adsorption of triclosan as model system was studied to qualify activated carbon sorbents recycled from gas masks (civilian gas mask GP5). The triclosan equilibrium concentration was measured spectrophotometrically, the morphology of the activated carbon characterized by scanning electron microscopy, and the amount of the adsorbed triclosan on the activated carbon quantified by a mass balance method. Experimental isotherms were fitted by Langmuir, Freundlich and Sips adsorption models. It was obtained that the contact time is a crucial sorption parameter that provides information on the optimum adsorption efficiency. It was shown that the maximum efficiency of GP5 (88%) is obtained after 10days of adsorption at a maximal concentration of triclosan and carbon loading 1mg/l. No significant adsorption efficiency differences were measured after 5 and 10days of adsorption. The non-linear Sips isotherm, a combined Freundlich–Langmuir model, provides suitable fitting results. The observed remarkable adsorption capacity of activated carbon (GP5) towards triclosan adsorption (∼85mg/g) makes it a viable solution for wastewater treatment.