Advanced treatment for arthritic diseases based on the capture and inactivation of interleukin-6 by biofunctionalized polymeric nanoparticles

Arthritic diseases, such as osteoarthritis and rheumatoid arthritis, are associated with synovium inflammation (synovitis). Several pro-inflammatory cytokines, especially tumor necrosis factor-α (TNFα) and interleukins (IL), are important mediators of inflammation and articular cartilage destruction, supporting a potential possibility of anticytokine therapy in these diseases. IL-6 is one of the key regulators of the inflammatory response. Thus, human monoclonal antibodies against IL-6 may prevent its action, and consequently reduce inflammation after intra-articular (IA) injection. Indeed, several clinical trials have already demonstrated positive outcomes over disease progression. Although these treatments are very attractive, they are associated with limited efficacy because of the rapid clearance of antibodies by the synovium. A solution to overcome this problem is using nanoparticles (NPs) as a substrate to protect and extend the action of the antibodies. Natural-derived polymers, like chitosan (Ch) and hyaluronic acid (HA), are biocompatible and biodegradable polysaccharides, being HA a natural component of the extracellular matrix of articular cartilage. Therefore, biodegradable polymeric NPs represent a good candidate for IA administration. In the present work we propose natural biodegradable polymeric NPs biofunctionalized with immobilized antibodies that selectively capture and inactivate the pro-inflammatory cytokine IL-6, reducing synovium inflammation. Ch-HA NPs were successfully prepared by polyelectrolyte complexation and further stabilized through carbodiimide chemistry (ethyl(dimethylaminopropyl) carbodiimide (EDC)/Nhydroxysuccinimide (NHS)). The particle size and zeta potential of the NPs were optimized. Stable NPs with 121.8 ± 2.4 of particle diameter, 0.11 ± 0.01 of polydispersity index and +25.12 ± 1.86 mV of zeta potential were produced with 0.25 mg/mL of initial polymers concentrations, at pH 5 and with 50/200 mM of EDC/NHS concentration. The anti-IL-6 antibody was immobilized at the surface of Ch-HA NPs. After determining the maximum antibody immobilization ability (7 µg/mL), the capacity to capture the recombinant IL-6 was evaluated. The efficacy was around 94-97%. Biological assays demonstrated not only the cytocompatibility of the produced NPs with human articular chondrocytes (hACs) (Fig 1) and human macrophages, but also the benefits of the capture and inactivation of IL-6 after stimulation with monocyte-derived macrophage conditioned medium. In conclusion, it is foreseeable that these NPs will overcome the limitations of the abovementioned treatments, since such NPs will increase the therapeutic efficacy due to their subcellular size, non-toxicity and high stability, being a promising approach for the local and sustained treatment of arthritic diseases.info:eu-repo/semantics/publishedVersio

Evolutionary algorithms for static and dynamic optimization of fed-batch fermentation processes

In this work, Evolutionary Algorithms (EAs) are used to control a recombinant bacterial fed-batch fermentation process, that aims at producing a bio-pharmaceutical product. In a first stage, a novel EA is used to optimize the process, prior to its start, by simultaneously adjusting the feeding trajectory, the duration of the fermentation and the initial conditions of the process. In a second stage, dynamic optimization is proposed, where the EA is running simultaneously with the fermentation process, receiving information regarding from the process, updating its internal model, reaching new solutions that will be used for online control

Multi-level dynamic modeling in biological systems : application of hybrid Petri nets to network simulation

The recent progress in the high-throughput experimental technologies allows the reconstruction of many biological networks and to evaluate changes in proteins, genes and metabolites levels in different conditions. On the other hand, computational models, when complemented with regulatory information, can be used to predict the phenotype of an organism under different genetic and environmental conditions. These computational methods can be used for example to identify molecular targets capable of inactivating a bacterium and to understand its virulence factors. This work proposes a hybrid metabolic-regulatory Petri net approach that is based on the combination of approximate enzyme-kinetic rate laws and Petri nets. A prototypic network model is used as a test-case to illustrate the application of these concepts in Systems Biology.This work was partially supported by post-doctoral grant by Fundacao para a Ciencia e a Tecnologia (FCT) (SFRH/BPD/80784/2011), project PneumoSyS - A Systems Biology approach to the role of pneumococcal carbon metabolism in colonization and invasive disease (FCT contract: PTDC/SAU-MII/100964/2008) and by FCT (INESC-ID multiannual funding) through the PIDDAC program funds

Synthesis of heterocyclic compounds by radical electrochemical approach in environmentally friendly media

Radical cyclisation is rapidly becoming an important method for the formation of cyclic systems. Hence, some electrochemical results obtained in the study of electroreductive intramolecular cyclisation of ethyl 2-bromo-3-(3,4-methylenedioxophenyl)-3-(propargyloxy)propanoate (1a), [1-bromo-2-methoxy-2-(prop-2’-ynyloxy) ethyl] benzene (1b) and 1-[2-bromo-2-phenyl-1-(prop-2’-ynyloxy)ethyl]-4-methoxybenzene (1c) promoted by (1,4,8,11-tetramethyl-1,4,8,11-tetraazacyclotetra-decane)nickel(I), [Ni(tmc)]+, electrogenerated at glassy carbon cathodes in ethanol, ethanol / water and microemulsions made with cationic and anionic surfactants are presented. The results obtained indicate that the reaction proceeds via cleavage of the carbon–bromine bond to form a radical intermediate that undergoes cyclization on the unsaturated C-C bond to afford the substituted tetrahydrofurans.Fundação para a Ciência e a Tecnologia (FCT

Testing natural biomaterials in animal models

Animal models have been extensively developed in the last decades in biomedical field. Their use has shown particular relevance in fields such as cell biology, genetics, anatomy and development, biochemistry, infection and immunity, cancer research, drugs and vaccine development, tissue engineering and regenerative medicine. The induced host tissue trauma and the inflammatory process resulting from the implantation of a medical device are of utmost importance for a successful outcome. Features of a chronic inflammation are usually attributed to the host response towards the implant, while an early acute inflammatory response is mainly endorsed by the implantation procedure. Animal testing comprises a midway step between in vitro studies and human clinical trials, which precede real clinical application. As every animal model has its advantages and disadvantages, a comprehensive analysis of each available species needs to be conducted when planning an animal study.(undefined)info:eu-repo/semantics/publishedVersio

Impact of MOF-5 on pyrrolidinium-based poly(ionic liquid)/ionic liquid membranes for biogas upgrading

PostprintBearing in mind that Metal Organic Frameworks (MOFs) have remarkable CO2 adsorption selectivity and Mixed Matrix Membranes (MMMs) have been identified as potential solution for advancing the current state of the art of membrane separation technology, this work investigates the effect of combining a MOF, with high adsorption properties towards CO2 when compared to CH4 (MOF-5), with a blend of poly(ionic liquid)/ionic liquid (PIL/IL) for biogas upgrading. The blend system consisted of a pyrrolidinium-based PIL, poly([Pyr11][Tf2N]), and a free imidazolium-based IL, [C2mim][BETI]. The MOF-5 was incorporated at different loadings (10, 20, 30 wt%), and MMMs were prepared by solvent evaporation technique and characterized by diverse techniques (FTIR, SEM, TGA, puncture tests, water contact angle and single gas transport). The results showed that the free IL is miscible with the PIL, while MOF-5 particles were uniformly dispersed into the PIL/IL matrix. The formed PIL/IL/MOF-5 membranes revealed suitable thermal stability (Tonset up to 656 K) for biogas upgrading processes, but a loss of mechanical stability was found after the incorporation of MOF-5, and thus more rigid and fragile membranes were obtained. Besides, increasing MOF-5 content in the MMMs resulted in improved CO2 permeability. At 30 wt% of MOF-5 loading the CO2 permeability increased 133% when compared to that of the pristine PIL/IL membrane, while the ideal selectivity CO2/CH4 decreases. It was possible to demonstrate the relevance of studying different components within the polymeric matrix in order to assess not only thermal, mechanical and chemical properties, but also gas transport response.This work was supported by FCT (Fundação para Ciência e a Tecnologia) through the project PTDC/CTM-POL/2676/2014 and the Associate Laboratory for Green Chemistry - LAQV (UID/QUI/50006/2019). Ana R. Nabais and Luísa A. Neves are grateful to FCT/MCTES for their PhD grant (SFRH/BD/136963/2018) and FCT Investigador Contract (IF/00505/2014), respectively. This project has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement no. 745734

Reactions radicalaires dans des milieux protiques et aprotiques : comparaisons entre les cyclisations reductives electrochimiques catalysees par des complexes du nickel

The electrochemical reductive cyclisation of unsaturated organic halides in the presence of Ni(II) complexes as catalysts was examined in aprotic solvents such as DMF and in protic solvents such as ethanol, butanol or ethanolewater mixtures. The presence of the alcohol media enhanced the rate of recycling of the catalytic species.La cyclisation réductive électrochimique d’halogénures insaturés en présence de complexes de Ni(II) comme catalyseurs a été examinée dans des solvants comme le DMF et dans des solvants protiques comme l’éthanol, le butanol ou des mélanges éthanol-eau. La présence du milieu alcoolique augmente la vitesse du recyclage des espèces catalytiques.Fundação para a Ciência e a Tecnologia (FCT

Development of suported ionic liquids for the purification of antileukemic drugs

L-asparaginase (LA) is an antileukemic biopharmaceutical of current high-cost. LA is produced via fermentation and its purification usually comprises precipitation, liquid- liquid extraction and chromatography techniques [1]. This work aims to develop sustainable technologies to purify LA. Functionalized nanomaterials, namely supported ionic liquids (SILs), are used as cost-effective purification techniques for the target enzyme. Initially, the synthesis and modification of SILs was performed. Different SILs were obtained and used for the purification of LA. Commercial LA was used for the first purification tests, in order to understand the behavior of the enzyme in contact with the nanomaterial. Experimental conditions, such as pH, and material/LA ratio, contact time were optimized. LA activity was quantified by Nessler reaction [2]. The first results reveal a total adsorption of LA by the SILs with a recovered activity reaching 90% depending on the SILs functionalization/ treatment. The modified SILs are shown to be very promising nanomaterials for the purification of LA. The LA was easily attached to SILs by adsorption under mild conditions. SILs supports can be a real alternative for a single step immobilization/purification of LA.publishe

Adesão à Terapêutica Alimentar Prescrita em Diabéticos Tipo1

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