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Operation conditions to recover and quantify active compounds from Perle Hallertau (PH) and Nuggets (N) hops, following a sequential combination of ultrasound and microwave assisted ethanolic extraction were studied. Ultrasound treatment (55 °C, 60% ethanol, 30 minutes) allowed extracts with phenolic contents of 160.43 mg GAE/g extract and ABTS radical scavenging capacity equivalent to 400.24 mg Trolox/g extract for PH. Optimal conditions for N were found at 55 °C, using 60% ethanol for 120 minutes (98.80 mg GAE/g extract) and 55 °C, 40% ethanol for 180 min (370.00 mg Trolox/g extract). The remaining solids were further processed under ethanolic microwave extraction (130–190 °C, 5 minutes) with the optimal results obtained at 190 °C for PH (77.11 and 98.67 mg GAE/g extract) and N (226.96 and 368.72 mg Trolox/g extract). Results obtained provided hops pellets as a rich source of active compounds that could be suitable for food and no food industry applications Ministerio de Ciencia, Innovación y Universidades | Ref. RYC 2018-024454-I

This study focusses on the valorization of the red seaweed Mastocarpus stellatus, collected in the Northern coast of Portugal, as a natural source of high value compounds due to its beneficial properties. An annual monitoring of the seaweed was performed by determining its lipids, ash, carbohydrates, phycobiliproteins, total phenolic compounds, antioxidant capacity and carrageenan from three different rocky shores located in the north of Portugal. The results showed a seasonal and spatial variability of the studied compounds between October 2018 and September 2019 depending on the climatic variables of temperature, precipitation, and solar radiation. The most productive season coincided with the warmest months, except for carbohydrates and phycobiliproteins, which were promoted in the colder season. The spatial variation also could be explained by the proximity to water channel discharges at the sampling sites. Complementary studies on the carrageenan fraction were conducted in one of the shores due to the high biopolymer content, to determine their carrageenan proportion between the summer and winter period and establish their rheological capabilities for the formulation of gelling matrices. The extracted biopolymers exhibited typical structural and viscoelastic characteristics of kappa/iota-hybrid carrageenans, suggesting notably differences depending on the harvest season, which is critically relevant to define future applications. Agencia Estatal de Investigación | Ref. RYC2018-024454-I Xunta de Galicia | Ref. ED431F 2020/01 Fundação para a Ciência e a Tecnologia | Ref. UIDP/04423/2020 Fundação para a Ciência e a Tecnologia | Ref. UIDB/04423/2020 Universidade de Vigo/CISUG

Polarized hydroxyapatite (p-HAp), a calcium phosphate catalyst obtained at high temperature under intense electric field, has been used for the synthesis of ammonium starting from N2 and liquid water at low pressure (

The enhanced catalytic activity of permanently polarized hydroxyapatite, which is achieved using a thermally stimulated polarization process, largely depends on both the experimental conditions used to prepare crystalline hydroxyapatite from its calcium and phosphate precursors and the polarization process parameters. A mineral similar to brushite, which is an apatitic phase that can evolve to hydroxyapatite, is found at the surface of highly crystalline hydroxyapatite. It appears after chemical precipitation and hydrothermal treatment performed at 150 °C for 24 h followed by a sinterization at 1000 °C and a polarization treatment by applying a voltage of 500 V at high temperature. Both the high crystallinity and the presence of brushite-like phase on the electrophotocatalyst affect the nitrogen and carbon fixation under mild reaction conditions (95 °C and 6 bar) and the synthesis of glycine and alanine from a simple gas mixture containing N2, CO2, CH4 and H2O. Thus, the Gly/Ala ratio can be customized by controlling the presence of brushite on the surface of the catalyst, enabling to develop new strategies to regulate the production of amino acids by nitrogen and carbon fixation.

Background Kiwifruit is a worldwide appreciated fruit with significant economic and nutritionally importance. Its industrial processing generates high amount of waste, which currently exhibits low market value being mostly employed as fertilizer. Nevertheless, culled kiwifruit, underused peels, seeds or pruning remains (i.e. leaves and twinges) would be an attractive source of bioactive compounds with prospective applications in the food and non-food sectors. Kiwifruit and the corresponding wastes display a notable number of healthy features with relevant benefits for consumer's health (e.g. strong antioxidant, anti-inflammatory, and protective against heart diseases, cancer, diabetes, or central nervous system diseases). Scope and approach Overall, this work explores a new approach about nutritional composition of kiwifruit and its different by-products fractions. This review also provides an outline on the bioactive compounds found in kiwifruit, products and by-products. In parallel, it offers an updated vision of the impact of cultivation and processing conditions on their bioactive characteristics. Moreover, the positive aspects of selecting eco-friendly extraction techniques as well as green and efficient treatments to provide high value added products from underutilized kiwifruit wastes is pointed out. Key findings and conclusions Kiwifruit and its by-products are natural resources which present several nutraceutical phytocompounds with potential commercial applications. The valorization of these waste-to-value food can be addressed through environmentally friendly extraction procedures which allow recovery of target bioactive components to enrich novel food and non-food products.

The synthesis of hydroxyapatite (HAp) with different shapes and sizes has attracted increasing attention because the applicability of this ceramic material depends on structure-properties relationships (i.e., the dimensions and morphology of HAp crystals determine properties such as the bioactivity and mechanical strength). Although different synthetic routes based on the addition of surfactants, organic modifiers, or dispersants have been proposed to control the growth of HAp crystals, many efforts are being devoted to simplify the whole process using simple parameters such as pH. However, the control of the morphology is still poor and shows low reproducibility. In this work, a new additive-free synthetic route, which is based on the hydrothermal method and the utilization of nonaqueous solvents, is proposed. The influence of the synthesis parameters such as pH, concentration of starting solutions, and the solvent on relevant features such as phase purity, crystallinity, crystallite size, and morphology has been examined using spectroscopic techniques, X-ray diffraction, and scanning electron microscopy. As a consequence, this work presents an easy and robust method based only on the use of organic solvent and the control of the pH that produces pure and crystalline HAp with a controlled shape and size. This method has been used to elucidate some of the key aspects of the crystal growth mechanism and to synthesize HAp crystals with different and well-defined shapes (e.g., belts, rods, flakes needle-like, or polymorph) and sizes, in a reproducible way.

This study deals with the valorisation of tea leaves from the agricultural pruning remains using clean technologies with low environmental impact. Leaves with null commercial interest were dismissed during the pruning season, however with a sequential combination of microwave hydrodiffusion and gravity (MHG), ultrasound-assisted (US) and pressurized hot water (AH) extractions was possible to recover bioactive compounds. An initial pre-treatment was performed using MHG at selected irradiation powers to facilitate the liberation of antioxidants through the cell membrane. The solid phases were processed by three ways: i) ultrasound bath system (80 °C, 80 KHz, from 15 to 60 min); ii) AH technology (non-isothermal treatment with maximum temperature from 140 to 200 °C); and iii) a sequence of the three aforementioned technologies. Both raw material and liquors were analysed to determine fundamental physicochemical, phytochemical or biological properties. Leaves debris showed positive results in bioactive compounds with a maximum value of total phenolic around 130 mg GAE/g extract in the sequential combination. Antioxidant capacity ranged from 0.3 to 0.9 g Trolox/g extract. The combination of green extraction technologies guarantees an efficient recovery of bioactive compounds, creating the possibility of new products that improve the wellbeing.

This work deals with the study of tea stalks from pruning debris using environmental friendly extraction technology to offer new healthy properties. In the manufacturing tea industry, tea trees require to be pruned every year and most of their remains are discarded as a waste with no economic value. Microwave aqueous extraction and pressurized hot water extraction process (autohydrolysis) were used to recover bioactive compounds from the tea branches. Operating at a fixed solid: liquid ratio (1:15), the effect of the maximum heating temperatures from 140 to 220 °C was studied. Liquid extracts were analysed for total phenolic, oligosaccharides, protein, mineral and heavy metals content, as well as for antioxidant capacity. The antitumoral possibilities were also determined for selected samples. The obtained results indicated that both processes could be used as an alternative to recover bioactive compounds from tea wastes, although microwave-assisted extraction allowed saving time when compared with autohydrolysis processing. The temperature exhibited a relevant effect on the total phenolic content and antioxidant capacity, decreasing with the microwave treatment and increasing with the autohydrolysis temperature. The obtained extracts could be adequate for incorporation in food and non-food fields., Graphical abstract Image 1, Highlights • Tea pruning remains were valorised using green extractions by microwave (MW) and autohydolysis (AH). • MW and AH were efficient technologies to recover bioactive compounds. • Values above 40 mg gallic acid equivalents/g extract and 0.10 g Trolox/g extract were identified. • Future applications in cosmetics, pharmacy or food industries should be explored.

The design of catalysts with controlled selectivity at will, also known as catalytic plasticity, is a very attractive approach for the recycling of carbon dioxide (CO2). In this work, we study how catalytically active hydroxyapatite (HAp) and brushite (Bru) interact synergistically, allowing the production of formic acid or acetic acid depending on the HAp/Bru ratio in the catalyst. Raman, wide angle X-ray scattering, X-ray photoelectron spectroscopy, scanning electron microscopy and electrochemical impedance spectroscopy studies, combined with an exhaustive revision of the crystalline structure of the catalyst at the atomic level, allowed to discern how the Bru phase can be generated and stabilized at high temperatures. Results clearly indicate that the presence of OH– groups to maintain the crystalline structural integrity in conjunction with Ca2+ ions less bonded to the lattice fixate carbon into C1, C2 and C3 molecules from CO2 and allow the evolution from formic to acetic acid and acetone. In this way, the plasticity of the HAp-Bru system is demonstrated, representing a promising green alternative to the conventional metal-based electrocatalysts used for CO2 fixation. Thus, the fact that no electric voltage is necessary for the CO2 reduction has a very favorable impact in the final energetic net balance of the carbon fixation reaction.

Aristotelia chilensis is a plant rich in phenolics and other bioactive compounds. Their leaves are discarded as waste in the maqui berry industry. A new application of these wastes is intended by the recovery of bioactive compounds using pressurized hot water extraction with conventional or microwave heating. Both technologies have been selected for their green character regarding the type of solvent and the high efficiency in shorter operation times. Extractions were performed in the temperature range 140–200 °C with a solid/liquid ratio of 1:15 (w:w). The extracts’ total phenolic content, antioxidant capacity, and saccharides content obtained with both heating methods were measured. Additionally, the thermo-rheological properties of the gelling matrix enriched with these extracts were analyzed. Optimum conditions for lyophilized extracts were found with conventional heating, at 140 °C and 20 min extraction, 250.0 mg GAE/g dry extract and 1321.5 mg Trolox/g dry extract. Close to optimum performance was achieved with microwave heating in a fraction of the time (5 min) at 160 °C (extraction), yielding extracts with 231.9 mg GAE/g dry extract of total phenolics and antiradical capacity equivalent to 1176.3 mg Trolox/g dry extract. Slightly higher antioxidant values were identified for spray-dried extracts (between 5% for phenolic content and 2.5% for antioxidant capacity). The extracts obtained with both heating methods at 200 °C contained more than 20% oligosaccharides, primarily glucose. All the formulated gelling matrices enriched with the obtained extracts displayed intermediate gel strength properties. The tested technologies efficiently recovered highly active antioxidant extracts, rich in polyphenolics, and valuable for formulating gelling matrices with potential applicability in foods and other products.

Background Hops are mostly processed for the extraction of compounds conferring aroma and bitterness to beer. A more controllable addition of these properties can be achieved when solvent hop extracts are produced and added in the desired brewing stage. Scope and approach This review presents a survey on the research and patents on emerging extraction technologies and the importance of the solvent and operational conditions on the composition of hop extracts, destined for the brewing industry, but also for the production of novel ingredients with potential for nutraceuticals. Key findings and conclusions For the preparation of extracts, the efficiency of conventional solvents has been progressively improved by the assistance with intensified technologies and particularly by the use of greener solvents, a trend where this industry has been pioneer. The design of multistage multiproduct processes for the integral utilization of hops following a biorefinery approach is also addressed.

The insertion of an implant in the body of a patient raises the risk of a posterior infection and formation of a biofilm, which can have critical consequences on the patient's health and be associated with a high sanitary cost. While antibacterial agents can be used to prevent the infection, such a strategy is time-limited and causes bacteria resistance. As an alternative to biochemical approaches, we propose here to use light-induced local hyperthermia with plasmonic nanoparticles. This strategy is implemented on surgical meshes, extensively used in the context of hernia repairing, one of the most common general surgeries. Surgical meshes were homogeneously coated with gold nanorods designed to efficiently convert near-infrared light into heat. The modified mesh was exposed to a biofilm of Staphylococcus aureus ( S. aureus) bacteria before being treated with a train of light pulses. We systematically study how the illumination parameters, namely fluence, peak intensity and pulse length, influence the elimination of attached bacteria. Additionally, fluorescence confocal microscopy provides us some insight on the mechanism involved in the degradation of the biofilm. This proof-of-principle study opens a new set of opportunities for the development of novel disinfection approaches combining light and nanotechnology.

Owing to their unique combination of chemical and physical properties, inorganic nanoparticles show a great deal of potential as suitable agents for early diagnostics and less invasive therapies. Yet, their translation to the clinic has been hindered, in part, by the lack of non-invasive methods to quantify their concentration in vivo while also assessing their effect on the tissue physiology. In this work, we demonstrate that diffuse optical techniques, employing near-infrared light, have the potential to address this need in the case of gold nanoparticles which support localized surface plasmons. An orthoxenograft mouse model of clear cell renal cell carcinoma was non-invasively assessed by diffuse reflectance and correlation spectroscopies before and over several days following a single intravenous tail vein injection of polyethylene glycol-coated gold nanorods (AuNRs-PEG). Our platform enables to resolve the kinetics of the AuNR-PEG uptake by the tumor in quantitative agreement with ex vivo inductively coupled plasma mass spectroscopy. Furthermore, it allows for the simultaneous monitoring of local tissue hemodynamics, enabling us to conclude that AuNRs-PEG do not significantly alter the animal physiology. We note that the penetration depth of this current probe was a few millimeters but can readily be extended to centimeters, hence gaining clinical relevance. This study and the methodology presented here complement the nanomedicine toolbox by providing a flexible platform, extendable to other absorbing agents that can potentially be translated to human trials.

Liver fibrosis is a major health problem with multiple associated complications, which, to date, has no effective treatment. Hepatic stellate cells are the main responsible cells for fibrosis formation; upon their activation, excess accumulation of extracellular matrix and collagen deposits occurs. The mitogen platelet-derived growth factor (PDGF) and its receptor β (PDGFRβ) play a major role in hepatic stellate cells activation and are, therefore, promising targets for antifibrotic therapies. Gold nanorods hold great potential for diseased liver treatments, since their passive hepatic accumulation enhances active targeting strategies, hence increasing therapeutic efficiency. In addition, gold nanorods have photothermal properties that, combined with specific cell delivery, can be exploited to induce localized near-infrared light-mediated thermal ablation. Here, we demonstrate that gold nanorods coated with anti-PDGFRβ specifically target activated hepatic stellate cells in vivo. Additionally, gold nanorods-PDGFRβ-mediated photothermal therapy decreases fibrosis, hepatic inflammation, and hepatocyte injury in the experimental model of CCl4-induced liver fibrosis in mice.

Native starch from potatoes and hybrid carrageenans from the red alga Mastocarpus stellatus have been used as gluten-free gelling matrices to obtain functional hydrogels. The enrichment of gelling matrices with antioxidant compounds from natural sources is an increasing market trend. In this context, this work is aimed at the formulation and thermo-rheological characterization of functional hydrogels using potato starch from agro-industrial waste and kappa–iota hybrid carrageenans extracted from the above seaweed, enriched with antioxidant compounds from different agro-industrial products, such as waste coming from the pruning of green tea and two varieties of hops used in the brewing industry. Environmentally friendly technologies such as microwave hydrodiffusion and gravity, microwave-assisted extraction, ultrasounds and autohydrolysis were used for the recovery of antioxidant compounds. The results point out that functional hydrogels based on potato starch and hybrid carrageenans with a wide range of viscoelastic features can be achieved, with the particularity of being suitable for people with celiac disease. The incorporation of selected antioxidant extracts from vegetable by-products involved the drop (about tenfold) of the viscous and elastic properties of the formulated gels. The sequential combination of the above treatments could even further expand the thermo-rheological properties of formulated hydrogels, with potential application in functional foodstuffs and novel gluten-free goods. Xunta de Galicia | Ref. ED431F 2020/01 Ministerio de Ciencia, Innovación y Universidades (España) | Ref. RTI2018-096376-B-I00 Ministerio de Ciencia, Innovación y Universidades (España) | Ref. RYC2018-024454-I

Hydroxyapatite (HAp) is a naturally occurring mineral form of calcium apatite of biomedical importance due to its similarity with human hard tissues in morphology and composition. Upon polarization at high temperature, applying 3 kV/cm at 1000¿°C, the resulting polarized HAp (p-HAp) exhibits enhanced catalytic behavior due charge accumulation at the interface. More specifically, p-HAp was found to catalyse the conversion of mixtures of CO2(g) and CH4(g) into low carbon organic molecules and into amino acids when N2(g) was added to the mixture. In this work, we report how p-HAp facilitates the conversion of CO2(g) mainly in ethanol by means of forming C-C coupled bonds on its activated surface. After evaluation of a wide range of experimental conditions, we evidence the production of formic acid, methanol and formaldehyde (C1 products); ethanol and acetic acid (C2 products); and acetone (C3 product) from CO2(g) using moderate reaction conditions. Moreover, optimization of the reaction parameters led to a significant increase towards ethanol.

Conversion of CO2 into valuable chemicals is not only a very challenging topic but also a socially demanding issue. In this work, permanently polarized hydroxyapatite obtained using a thermal stimulated polarization process is proposed as a highly selective catalyst for green production of ethanol starting from CO2 and CH4.

Fluorescence microscopy is the method of choice in biology for its molecular specificity and super-resolution capabilities. However, it is limited to a narrow z range around one observation plane. Here, we report an imaging approach that recovers the full electric field of fluorescent light with single-molecule sensitivity. We expand the principle of digital holography to fast fluorescent detection by eliminating the need for phase cycling and enable three-dimensional (3D) tracking of individual nanoparticles with an in-plane resolution of 15 nm and a z-range of 8 mm. As a proof-of-concept biological application, we image the 3D motion of extracellular vesicles (EVs) inside live cells. At short time scales (, Science Advances, 6 (45), ISSN:2375-2548

The electrophotocatalytic synthesis of Glycine and Alanine from a simple gas mixture containing N2, CO2, CH4 and H2O under mild reaction conditions (95 °C and 6 bar) was recently developed using a catalyst formed by permanently polarized hydroxyapatite, which is achieved using a thermally stimulated polarization process, coated with two layers of aminotris(methylenephosphonic acid) (ATMP) separated by an intermediate layer of zirconyl chloride (ZC). This work reports the optimization of the ATMP- and ZC-coating content by examining the influence of their concentration of each component in each layer on the structural and electrochemical properties of the catalyst. After exhaustive analyses, such properties have been related with the efficiency of the catalysts prepared using different ATMP- and ZC-concentrations to yield Gly and Ala amino acids by fixing nitrogen from N2 and carbon from CO2 and CH4. Results show that, although the concentrations of ATMP and ZC in the first and the intermediate layers are important, the third layer plays a predominant role as is responsible of the apparition of supramolecular structures on the surface and the capacitive behavior of the coating.

Design of hydroxyapatite (HAp) with customized electrical properties is of special interest for developing technological and biomedical applications with new improved functionalities. Polarized HAp, which is obtained by applying an external electric voltage at high temperature, has been successfully shown to be an alternative to doped-HAp that is limited by the biocompatibility of the dopants used. However, many aspects about such new material remain scarcely studied, as for example the relationship between the polarization conditions and the resultant electrical enhancement, hinder a solid progress in its application. In this work, polarized HAp has been extensively characterized using electrochemical impedance spectroscopy by means of proposing a unified electrical equivalent circuit model with physical sense. This allows to explain the properties of such material by separating the bulk and the interface contributions. Moreover, the limits of the polarization mechanism have been explored, enabling a precise control on the electrical resistivity of polarized HAp above or below the intrinsic resistivity of nonpolarized HAp. Overall, necessary insights on the polarization treatment have been reported, opening an appealing avenue for generating new biomedical and technological applications based on dopant-free polarized HAp.

The goal of this paper is the valorisation of the Chondrus crispus (Rhodophyta) wastes generated during the industrial extraction of hybrid carrageenan using microwave technology of a temperature range from 140 to 200 °C, using water as a solvent. Mineral and phenolic content, antioxidant capacity and thermorheological features of the recovered hybrid carrageenans were determined. Results indicated the suitability for the recovery of high valuable compounds from these algae residues, after the optimization of the extraction conditions. At microwave processing conditions (160 °C, 5 min), it is still possible to recover around 10% of semi-refined hybrid carrageenan from the industrial raw material as well as to achieve the maximum value of total phenolic compounds of 12.8 mg GAE/g extract and antioxidant capacity of 51.70 mg Trolox/g extract. Extracted biopolymers exhibited thermomechanical features, in terms of viscoelastic moduli or gelling and melting temperatures, comparable to their commercial counterparts.

Biosensing based on optical micro- and nanoresonators integrated in a microfluidic environment is a promising approach to lab-on-a-chip platforms capable of detecting low concentrations of analytes...

Noble metal nanoparticles (NPs) supporting localized surface plasmon resonances are widely used in the context of biotechnology as optical and absorption contrast agents with great potential applicability to both diagnostics and less invasive therapies. In this framework, it is crucial to have access to simple and reliable microscopy techniques to monitor the NPs that have internalized into cells. While dark field (DF) microscopy takes advantage of the enhanced NP scattering at their plasmon resonance, its use in cells is limited by the large scattering background from the internal cell compartments. Here, we report on a novel two-color dark field microscopy that addresses these limitations by significantly reducing the cell scattering contribution. We first present the technique and demonstrate its enhanced contrast, specificity and reliability for NP detection compared to a standard optical dark field. We then demonstrate its potential suitability in two different settings, namely wide-field parallel screening of circulating cells in microfluidic chips and high-resolution tracking of internalized NPs in cells. These proof of principle experiments show a promising capability of this approach with possible extension to other kinds of targeted systems like bacteria and vesicles.

Nanomedicine has emerged as a promising strategy to address some of the limitations of traditional biomedical sensing, imaging and therapy modalities. Its applicability and efficacy are, in part, hindered by the difficulty in both controllably delivering nanoparticles to specific regions and accurately monitoring them in tissue. Gold nanoparticles are among the most extensively used inorganic nanoparticles which benefit from high biocompatibility, flexible functionalization, strong and tunable resonant absorption, and production scalability. Moreover, their capability to enhance optical fields at their plasmon resonance enables local boosting of non-linear optical processes, which are otherwise very inefficient. In particular, two-photon induced luminescence (TPL) in gold offers high signal specificity for monitoring gold nanoparticles in a biological environment. In this article, we demonstrate that TPL microscopy provides a robust sub-micron-resolution technique able to quantify accumulated gold nanorods (GNRs) both in cells and in tissues. First, the temporal accumulation of GNRs with two different surface chemistries was measured in 786-O cells during the first 24 hours of incubation, and at different nanoparticle concentrations. Subsequently, GNR accumulation in mice, 6 h and 24 hours after tail vein injection, was quantified by TPL microscopy in biopsied tissue from kidney, spleen, liver and clear cell renal cell carcinoma (ccRCC) tumors, in good agreement with inductively coupled mass spectroscopy. Our data suggest that TPL microscopy stands as a powerful tool to understand and quantify the delivery mechanisms of gold nanoparticles, highly relevant to the development of future theranostic medicines.

The need for point-of-care devices able to detect diseases early and monitor their status, out of a lab environment, has stimulated the development of compact biosensing configurations. Whereas localized surface plasmon resonance (LSPR) sensing integrated into a state-of-the-art microfluidic chip stands as a promising approach to meet this demand, its implementation into an operating sensing platform capable of quantitatively detecting a set of molecular biomarkers in an unknown biological sample is only in its infancy. Here, we present an on-chip LSPR sensor capable of performing automatic, quantitative, and multiplexed screening of biomarkers. We demonstrate its versatility by programming it to detect and quantify in human serum four relevant human serum protein markers associated with breast cancer.

Owing to their unique chemical and physical properties, colloidal gold nanoparticles have prompted a wide variety of biocompatible nano-agents for cancer imaging, diagnosis and treatment. In this context, biofunctionalized gold nanorods (AuNRs) are promising candidates for light-induced hyperthermia, to cause local and selective damage in malignant tissue. Yet, the efficacy of AuNR-based hyperthermia is highly dependent on several experimental parameters; in particular, the AuNR morphology strongly affects both physical and biological involved processes. In the present work, we systematically study the influence of different structural parameters like the AuNR aspect ratio, length and molecular weight on in vitro cytotoxicity, cellular uptake and heat generation efficiency. Our results enable us to identify the optimum AuNR morphology to be used for in vivo hyperthermia treatment.

ObjectivosQuantificar a relevu00e2ncia clu00ednica dos sintomas que apresentam os pacientes com Dependu00eancia a Opiu00e1ceos (CIE 10 F11.22) e a sua variau00e7u00e3o com respeito aos afectados tambu00e9m por outro Trastorno Mental diferente.Material y mu00e9todosEstudam-se 185 pacientes incluu00eddos no Programa de Objetivos Intermu00e9dios de Bitarte com diagnu00f3stico de dependu00eancia a opiu00e1ceos (CIE 10 F11.22).Divide-se a amostra em dois grupos: A.- Um u00e9 constituu00eddo por pacientes com diagnu00f3stico u00fanico de dependu00eancia a opiu00e1ceos (CIE 10 F11.22) B.- Outro com pacientes que tu00eam associado outro diagnu00f3stico adicional (CIE 10; F20, F30, F40 o F60).Administram-se os testes: BDI II, Hamilton, Rosemberg, BIS 11 y SF 36.Mediante anu00e1lises estatu00edsticas (SPSS) comparamos o grupo que apenas apresenta patologia relacionada com substu00e2ncias, com os que u00e0 parte de esta, apresentam outra patologia psiquiu00e1trica associada.ConclusoesPodemos afirmar que, na amostra analisada no nosso Serviu00e7o os pacientes que apresentam comorbidade com os transtornos relacionados com substu00e2ncias tu00eam de forma estatisticamente significativa:Estado de u00e2nimo mais baixo, maior grau mu00e9dio de ansiedade, pior autoestima, su00e3o significativamente mais impulsivos, y percebem uma pior qualidade de vida que os que apresentam exclusivamente dependu00eancia a opiu00e1ceos

Like other hemeproteins, ferric hemoglobin (HbIII) not only can act as a catalyzer in the oxidation of substrates by H 2 O 2 but also can act alone as a H 2 O 2 reducer. In this paper, the kinetic mechanism of the reaction between HbIII and H 2 O 2 is studied, and a mathematical model is developed, which permits the determination of the kinetic constants from the absorbance measurement; the results indicate that each HbIII molecule is able to oxidize up to 5 H 2 O 2 molecules without external regeneration. Second, it is demonstrated that the optical properties of HbIII can also be used as an indicator of this oxidation reaction, and analytical methods can be developed using this approach. A method for H 2 O 2 peroxide determination is presented, which permits the analyte to be determined in a concentration range that depends on the HbIII concentration used and the measurement time, with an RSD better than 5%. In addition, methods for peracetic acid (PPA), the simultaneous determination of PAA + H 2 O 2 and the use of HbIII/H 2 O 2 as an indicator for oxidase enzymatic reactions (glucose/glucose oxidase) are outlined.

Label-free biosensing based on metallic nano-particles supporting localized surface plasmon resonances (LSPR) has recently received growing interest (Anker, J. N., et al. Nat. Mater. 2008, 7, 442-453). Besides its competitive sensitivity (Yonzon, C. R., et al. J. Am. Chem. Soc. 2004, 126, 12669-12676; Svendendahl, M., et al. Nano Lett. 2009, 9, 4428-4433) when compared to the surface plasmon resonance (SPR) approach based on extended metal films, LSPR biosensing features a high-end miniaturization potential and a significant reduction of the interrogation device bulkiness, positioning itself as a promising candidate for point-of-care diagnostic and field applications. Here, we present the first, paralleled LSPR lab-on-a-chip realization that goes well beyond the state-of-the-art, by uniting the latest advances in plasmonics, nanofabrication, microfluidics, and surface chemistry. Our system offers parallel, real-time inspection of 32 sensing sites distributed across 8 independent microfluidic channels with very high reproducibility/repeatability. This enables us to test various sensing strategies for the detection of biomolecules. In particular we demonstrate the fast detection of relevant cancer biomarkers (human alpha-feto-protein and prostate specific antigen) down to concentrations of 500 pg/mL in a complex matrix consisting of 50% human serum.

Optical biosensing based on gold nanoparticles supporting localized surface plasmoncs (LSPR) potentially offers great opportunities for compact, sensitive and low cost diagnostic devices. While last two decades have witnessed a diversity of nanoplasmonic systems with outstanding sensitivity, the implementation of LSPR sensing into a real analytical device is only at its infancy. In this context, we present here our latest advances in the optical, label free detection of biomolecules based on gold nanoantennas integrated into a state-of-the-art microfluidic platform. We first demonstrate the capability of our platform to detect low concentrations (

Peracetic acid (PAA) is selectively determined in the presence of hydrogen peroxide (H(2)O(2)) by using the self-indicating UV-Vis molecular absorption properties of catalase. The PAA reacts with the protein giving an intermediate (Cat-I) which is reduced back by the amino acid core surrounding the heme group. Since the original form of the enzyme and the Cat-I have different UV-Vis absorption properties, the absorbance changes can be used for PAA determination. The H(2)O(2)/catalase reaction is extremely fast so that neither Cat-I compound nor kinetic interferences are observed. The method permits PAA determination in the 5 × 10(-7) to 1.5 × 10(-5) M range, the reproducibility being between 1% and 10%. Using this method, PAA has been successfully determined in water samples treated with commercial PAA/H(2)O(2) biocides. A theoretical study has also been carried out for obtaining a mathematical model able to analytically describe the process.

Single-walled carbon nanotubes (SWCNTs) have been identified as a transporter for anti-cancer drugs, as they are capable of penetrating mammalian cell membranes and allow for a high drug loading due to their nanoscale dimensions and high aspect ratio. In addition, they can assist the targeting of therapeutic agents to the desired site of action by conjugation to antibodies or ligands of cancer cell surface receptors, which increases the effectiveness of the treatment and reduces side effects. In this work, we present a method for the triple functionalisation of oxidised SWCNTs with the anti-cancer drug doxorubicin, a monoclonal antibody, and a fluorescent marker at non-competing binding sites. The proposed methodology allows for the targeted delivery of the anti-cancer drug to cancer cells and the visualisation of the cellular uptake of SWCNTs by confocal microscopy. We show that the complex is efficiently taken up by cancer cells with subsequent intracellular release of doxorubicin, which then translocates to the nucleus while the nanotubes remain in the cytoplasm.

A new approach for glucose determination in blood based on the spectroscopic properties of blood hemoglobin (Hb) is presented. The biosensor consists of a glucose oxidase (GOx) entrapped polyacrylamide (PAA) film placed in a flow cell. Blood is simply diluted with bidistilled water (150:1, v:v) and injected into the carrier solution. When reaching the PAA film, the blood glucose reacts with the GOx and the resulting H(2)O(2) reacts with the blood Hb. This produces an absorbance change in this compound. The GOx-PAA film can be used at least 100 times. Lateral reactions of H(2)O(2) with other blood constituents are easily blocked (by azide addition). The linear response range can be fitted between 20 and 1200 mg dL(-1) glucose (R.S.D. 4%, 77 mg dL(-1)). In addition to the use of untreated blood, two important analytical aspects of the method are: (1) the analyte concentration can be obtained by an absolute calibration method; and (2) the signal is not dependent on the oxygen concentration. A mathematical model relating the Hb absorbance variation during the reaction with the glucose concentration has been developed to provide theoretical support and to predict its application to other compounds after changing the GOx by another enzyme. The method has been applied to direct glucose determination in 10 blood samples, and a correlation coefficient higher than 0.98 was obtained after comparing the results with those determined by an automatic analyzer. As well as sharing some of the advantages of disposable amperometric biosensors, the most significant feature of this approach is its reversibility.

Optical reagentless biosensors are one of the most promising alternatives for producing selective, sensitive and autonomous sensors for real life applications. These devices are based on the efficient use of the spectroscopic properties of bioreagents, mainly proteins, as transducers; avoiding in this way the use of chemical colorant/fluorophores which usually limit sensors performance. In this paper a brief state of the art of the bioreagents being used in biosensors as well as recent alternatives are discussed. The advantages of flavoenzymes and hemeproteins as the basis for reagentless biosensors are particularly stressed.

This paper demonstrates that glucose determination in blood can be done directly (without sample pretreatment) using a reagentless reversible biosensor based on the intrinsic spectroscopic properties of peroxidase (HRP). The biosensor, prepared by HRP and glucose oxidase entrapment in a polyacrylamide gel matrix, works in continuous mode, presents a linear response range from 1.5 × 10 −6 up to 5.5 × 10 −5 M and can be used for at least 750 measurements; in the best conditions (0.1 M pH 6 phosphate buffer, HRP and GOx amounts in the polymersation mixture for the sensor film preparation 0.0165 and 0.0010 g, respectively) the minimum samples rate is 30 h −1 . For glucose determination, blood is simply diluted in water (until haemolysis is completed) and fed into the sensor without a cleaning step between samples; the blood absorption is corrected in a simple way by working at a proper reference wavelength. The biosensor signals have been mathematically modeled in order to facilitate the design of sensors based on the same idea for other biochemical compounds.

During the reversible reaction between peroxidase (HRP) and H(2)O(2), several peroxidase intermediate species, showing different molecular absorption spectra, are formed which can be used for H(2)O(2) determination; when H(2)O(2) is generated in a previous enzymatic reaction, the substrate involved in this reaction can also be determined. On this basis, a new family of fully reversible reagentless optical biosensors containing HRP is presented; glucose determination is used as a model. The biosensor (which can be used for at least 6 months and/or more than 750 measurements) is prepared by HRP and glucose oxidase entrapment in a polyacrylamide gel matrix. A mathematical model (in which optical, kinetic and transport aspects are considered) relating the measured absorbance with the substrate concentration is also presented together with a simple methodology for characterization of this kind of biosensor. Regarding the optical model, the Kubelka-Mulk theory of reflectance does not give good results and the biosensors are better described by the Rayleigh theory of polymer solutions. Under working conditions, linear response ranges from 1.5x10(-6) to 3.0x10(-4)M glucose and CV was about 4%. This biosensor has been applied for glucose determination in fruit juices and synthetic serum samples without sample pretreatment.

Objectives Despite their various advantages, such as good esthetic properties, absence of mercury and adhesive bonding to teeth, modern dental composites still have some drawbacks, e.g., a relatively high rate of secondary caries on teeth filled with composite materials. Recent research suggests that microstructured biomaterials surfaces may reduce microbial adhesion to materials due to unfavorable physical material–microbe interactions. The objectives of this study were, therefore, to test the hypotheses that (i) different surface microstructures can be created on composites by a novel straightforward approach potentially suitable for clinical application and (ii) that these surface structures have a statistically significant effect on microbial adhesion properties. Methods Six different dental composites were initially tested for their suitability for microstructuring by polydimethylsiloxane (PDMS) templates. Each composite was light-cured between a glass slide and a microstructured PDMS template. The nano-hybrid composite Grandio Flow was the only tested composite with satisfying structurability, and was therefore used for the bacterial adhesion tests. Composites samples were structured with four different microstructures (flat, cubes, linear trapezoid structures, flat pyramids) and incubated for 4 h in centrifuged saliva. The bacterial adherence was then characterized by colony forming units (CFUs) and scanning electron microscopy (SEM). Results All four microstructures were successfully transferred from the PDMS templates to the composite Grandio Flow. The CFU-test as well as the quantitative analysis of the SEM images showed the lowest bacterial adhesion on the flat composite samples. The highest bacterial adhesion was observed on the composite samples with linear trapezoid structures, followed by flat pyramids and cubes. The microstructure of dental composite surfaces statistically significantly influenced the adhesion of oral bacteria. Significance Modifying the composite surface structure may be a clinically suitable approach to control the microbial adhesion and thus, to reduce the risk of secondary caries at dental composite restorations. Smaller composite surface structures may be useful for accomplishing this.

One of the difficulties in using optical biosensors based on enzymatic reactions is the immobilisation of the enzyme involved in the determination. A detailed study has been carried out of the various polymeric supports which could provide a potential alternative for the immobilisation of chemically modified enzymes. Before immobilisation, the enzyme is attached by means of a covalent bond to a fluorescent probe, which is optically active in the visible region. The main advantage of this covalent bond is that it is possible to follow the enzymatic reaction by fluorescence without the need for immobilising any further reagent (reagentless biosensors). The results indicate that the most stable and reproducible polymeric supports are derived from polyacrylamide obtained by UV photopolymerisation. The experiments have been carried out using chemically modified glucose oxidase as the model enzyme. The films thus obtained have a lifetime of at least two months, an RSD of 9.2%, and a linear range of 300 to 2000 mg L−1 of glucose. They are completely reversible by regeneration in an appropriate buffer solution.

In this paper an in depth study is presented of the use of the horseradish peroxidase (HRP) enzyme as a self-indicating biorecognition reagent in UV-vis molecular absorption spectrometry. The HRP/H2O2 reaction mechanism in the absence of an external substrate has been clarified, and the interaction between HRP and glucose oxidase (GOx) has been studied. It has been demonstrated that GOx can act as a substrate of HRP; in both cases the kinetic constants have been obtained and mathematical models have been developed. Second, the HRP/H2O2 reaction is used to follow a H2O2-producing enzymatic reaction, the glucose reaction with GOx being used as a model. As an application of this, two methodologies have been proposed for glucose determination: with or without previous incubation of glucose with GOx. In both cases mathematical models relating HRP absorbance changes to glucose concentration have been developed and tested; both methods have been optimized, analytically characterized, and tested for glucose determination in samples. The methodology described could be applied to other heme-proteins and to other H2O2-producing enzymatic reactions. The models permit the reaction constants to be calculated. From the analytical chemistry point of view the models allow the prediction of the method sensitivity for other analytes involved in this type of reaction if the kinetic constants are known and can be used in the design of optical sensors.

The use of non-viral vectors as delivery systems in gene therapy has been extensively studied recently owing to their advantages over viral vectors. Here, we propose a new gene delivery system based on the use of RNA-wrapped single-walled carbon nanotubes (SWCNTs) complexed with the cationic protein, protamine and the drug chloroquine. Protamine was selected as a cationic protein acting as bridge between negatively charged RNA-wrapped SWCNTs and plasmid DNA. Protamine also contains a nuclear localization signal which enhances the expression of the transfected gene. The drug chloroquine, a lysosomotropic compound which has been reported to increase the transfection efficiency, was attached to RNA-wrapped SWNTs by ionic interactions. The simultaneous delivery of the drug chloroquine with plasmid DNA clearly showed an enhanced gene delivery and expression. The levels of gene expression were quantified using the luciferase reporter gene as model. Optimal conditions for transfection and gene expression were obtained and cytoxicity of the carbon nanotube complexes measured. The optimal complexes were shown to efficiently deliver plasmid DNA for efficient gene expression and may thereby be useful as gene delivery systems for gene therapy.

The surface chemistry of gold nanoparticles (AuNPs) plays a critical role in the self-assembly of thiolated molecules and in retaining the biological function of the conjugated biomolecules. According to the well-established gold–thiol interaction the undefined ionic species on citrate-reduced gold nanoparticle surface can be replaced with a self-assembled monolayer of certain thiolate derivatives and other biomolecules. Understanding the effect of such derivatives in the functionalization of several types of biomolecules, such as PEGs, peptides or nucleic acids, has become a significant challenge. Here, an approach to attach specific biomolecules to the AuNPs (~14 nm) surface is presented together with a study of their effect in the functionalization with other specific derivatives. The effect of biofunctional spacers such as thiolated poly(ethylene glycol) (PEG) chains and a positive peptide, TAT, in dsRNA loading on AuNPs is reported. Based on the obtained data, we hypothesize that loading of oligonucleotides onto the AuNP surface may be controlled by ionic and weak interactions positioning the entry of the oligo through the PEG layer. We demonstrate that there is a synergistic effect of the TAT peptide and PEG chains with specific functional groups on the enhancement of dsRNA loading onto AuNPs.