Suchergebnisse

The effectiveness of most cancer targeted therapies is short-lived. Tumors often develop resistance that might be overcome with drug combinations. However, the number of possible combinations is vast, necessitating data-driven approaches to find optimal patient-specific treatments. Here we report AstraZeneca's large drug combination dataset, consisting of 11,576 experiments from 910 combinations across 85 molecularly characterized cancer cell lines, and results of a DREAM Challenge to evaluate computational strategies for predicting synergistic drug pairs and biomarkers. 160 teams participated to provide a comprehensive methodological development and benchmarking. Winning methods incorporate prior knowledge of drug-target interactions. Synergy is predicted with an accuracy matching biological replicates for >60\% of combinations. However, 20\% of drug combinations are poorly predicted by all methods. Genomic rationale for synergy predictions are identified, including ADAM17 inhibitor antagonism when combined with PIK3CB/D inhibition contrasting to synergy when combined with other PI3K-pathway inhibitors in PIK3CA mutant cells. ; We thank the Genomics of Drug Sensitivity in Cancer and COSMIC teams at the Wellcome Trust Sanger Institute for help with the preparation of the molecular data, Denes Turei for help with Omnipath, and Katjusa Koler for help with matching drug names across combination screens. We thank AstraZeneca for funding and provision of data to the DREAM Consortium to run the challenge, and funding from the European Union Horizon 2020 research (under grant agreement No 668858 PrECISE to J.S.R.), the Joint Research Center for Computational Biomedicine (which is partially funded by Bayer AG) to J.S.R., National Institute for Health Research (NIHR) Sheffield Biomedical Research Center, Premium Postdoctoral Fellowship Program of the Hungarian Academy of Sciences. M.G lab is supported by Wellcome Trust (102696 and 206194). ...

Engineered odorant-binding proteins (OBPs) display tunable binding affinities triggered by temperature alterations. We designed and produced two engineered proteins based on OBP-I sequence: truncated OBP (tOBP) and OBP::GQ20::SP-DS3. The binding affinity of 1-aminoanthracene (1-AMA) to these proteins revealed that tOBP presents higher affinity at 25°C (kd=0.45M) than at 37°C (kd=1.72M). OBP::GQ20::SP-DS3 showed an opposite behavior, revealing higher affinity at 37°C (kd=0.58M) than at 25°C (kd=1.17M). We set-up a system containing both proteins to evaluate their temperature-dependent binding. Our data proved the 1-AMA differential and reversible affinity towards OBPs, triggered by temperature changes. The variations of the binding pocket size with temperature, confirmed by molecular modelling studies, were determinant for the differential binding of the engineered OBPs. Herein we described for the first time a competitive temperature-dependent mechanism for this class of proteins. ; This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684). A.R. and F.G. thank FCT for funding their scholarships with the references SFRH/BPD/98388/2013 and SFRH/BD/114684/2016, respectively. T.G.C. thanks senior position funded by the European Union through the European Regional Development Fund (ERDF) under the Competitiveness Operational Program (COP-A1-A1.1.4-E nr.30/01.09.2016). C.S. thanks to the BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. Access to computing resources funded by the Project "Search-ON2: Revitalization of HPC infrastructure of UMinho" (NORTE-07-0162-FEDER-000086), co-funded by the North Portugal Regional Operational Programme (ON.2 - O Novo Norte), under the National Strategic Reference Framework (NSRF), through the European Regional Development Fund ...

Endolysins, which are peptidoglycan-degrading enzymes expressed during the terminal stage of the reproduction cycle of bacteriophages, have great potential to control Gram-pos- itive pathogens. This work describes the characterization of a novel endolysin (PlyPl23) encoded on the genome of Paenibacillus larvae phage phiIBB_Pl23 with high potential to control American foulbrood. This bacterial disease, caused by P. larvae, is widespread in North America and Europe and causes important economic losses in apiculture. The restric- tion to antibiotic residues in honey imposed by the EU legislation hinders its therapeutic use to combat American foulbrood and enforces the development of alternative antimicrobial methods. The new endolysin described herein has an N-acetylmuramoyl-L-alanine ami- dase catalytic domain and exhibits a broad-spectrum activity against common P. larvae genotypes. Moreover, the enzyme displays high antimicrobial activity in a range of pH that matches environmental conditions (pH between 5.0 and 7.0), showing its feasible applica- tion in the field. At pH 7.0, a concentration of 0.2 M of enzyme was enough to lyse 104 CFU.mL-1 of P. larvae in no more than 2 h. The presence of sucrose and of the substances present in the larvae gut content did not affect the enzyme activity. Interestingly, an increase of activity was observed when PlyPl23 was previously incubated in royal jelly. Furthermore, in vivo safety evaluation assays demonstrated that this enzyme is not toxic to the bee lar- vae. The present work describes for the first time an endolysin encoded in a P. larvae phage that presents high potential to integrate a commercial product to control the problematic American foulbrood. ; FCT—Foundation for Science and Technology strategic funding of UID/BIO/04469/2013 unit and the European Community fund FEDER, through Program COMPETE, under the scope of the Projects FCOMP-01-0124-FEDER-007025 (PTDC/AMB/68393/2006), PEst-OE/EQB/LA0023/2013, PEST-C/FIS/UI607/2013, RECI/BBB-EBI/0179/2012 ...

From the use of botanical plants in early human civilizations through synthetic chemistry and biotechnology, drug research has always passionate scientists creating exciting challenges to a large number of researchers from different fields, thus, promoting a collaborative effort between polymer scientists, pharmacologists, engineers, chemists and medical researchers. Worldwide, there is an increasing concern on health care that creates a major opportunity for development of new pharmaceutical formulations. Ageing populations worried about the quality of life in the older years are actively seeking for new, more effective and patient compliant drug delivery devices. This has been the driving force for the continuous growth of the research made on delivery devices, which has become a powerful technique in health care. It has been recognized for long that simple pills or injections may not be the suitable methods of administration of a certain active compound. These medications present several problems and/or limitations, like poor drug bioavailability and systemic toxicity, derived essentially from pharmacokinetic and other carrier limitations and low solubility of the drugs in water. Therefore and to overcome these drawbacks, clinicians recommend frequent drug dosing, at high concentrations, in order to overcome poor drug bioavailability but causing a potential risk of systemic toxicity. Polymer science has open new strategies for drug delivery systems. This Chapter overviews of possible strategies involving polymer modification and processing for controlled drug delivery and drug delivery in tissue engineering. ; European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement number REGPOT-CT2012-316331-POLARIS and from the project "Novel smart and biomimetic materials for innovative regenerative medicine approaches" RL1 -ABMR-NORTE-01-0124-FEDER-000016 cofinanced by North Portugal Regional Operational Programme (ON.2 –O Novo Norte), under the National Strategic Reference Framework (NSRF), ...

Background Half of human cancers harbour TP53 mutations that render p53 inactive as a tumor suppressor. As such, reactivation of mutant (mut)p53 through restoration of wild-type (wt)-like function represents one of the most promising therapeutic strategies in cancer treatment. Recently, we have reported the (S)-tryptophanol-derived oxazoloisoindolinone SLMP53-1 as a new reactivator of wt and mutp53 R280K with in vitro and in vivo p53-dependent antitumor activity. The present work aimed a mechanistic elucidation of mutp53 reactivation by SLMP53-1. Methods and results By cellular thermal shift assay (CETSA), it is shown that SLMP53-1 induces wt and mutp53 R280K thermal stabilization, which is indicative of intermolecular interactions with these proteins. Accordingly, in silico studies of wt and mutp53 R280K DNA-binding domain with SLMP53-1 unveiled that the compound binds at the interface of the p53 homodimer with the DNA minor groove. Additionally, using yeast and p53-null tumor cells ectopically expressing distinct highly prevalent mutp53, the ability of SLMP53-1 to reactivate multiple mutp53 is evidenced. Conclusions SLMP53-1 is a p53-activating agent with the ability to directly target wt and a set of hotspot mutp53. General Significance This work reinforces the encouraging application of SLMP53-1 in the personalized treatment of cancer patients harboring distinct p53 status. ; European Union (FEDER funds through Programa Operacional Factores de Competitividade – COMPETE) and National Funds (FCT/MEC, Fundação para a Ciência e Tecnologia and Ministério da Educação e Ciência) through the projects UID/QUI/50006/2019, COMPETE 2020 (POCI-01-0145-FEDER-006684/POCI-01-0145-FEDER-007440) and the BioTecNorte operation (NORTE-01-0145-FEDER-000004), (3599-PPCDT) PTDC/DTP-FTO/1981/2014 – POCI-01-0145-FEDER-016581 and UID/QUI/0081/2013; the Italian Association for Cancer Research, AIRC (IG#5506 to G.F.), Compagnia S. Paolo, Turin, Italy (Project 2017.0526 to G.F.) and Ministry of Health, (Project 5 × 1000, 2013 and 2015; ...

Current biomedical research has highlighted the potential use of medicinal plant derivatives as advanced suitable biomaterials due to their biological properties. Acemannan (ACE) is the main polysaccharide of Aloe vera (AV) leaves with many reported biological properties, including immunomodulatory activity, antibacterial action, wound healing induction, and cytocompatibility. These properties suggest that ACE is an appealing material for the development of 3D structures for biomedical purposes. In this study, ACE was modified by a methacrylation reaction using methacrylic anhydride. Further 3D sponges were developed by processing modified ACE by photocrosslinking using UV irradiation and the freeze-drying technique. Both FTIR and 1H NMR spectroscopy confirmed the effectiveness of the functionalization. SEM and m-CT revealed that the cross-sections of the methacrylated acemannan (MACE) sponges have open macropores and a narrow pore size distribution within them. We hypothesized that the intrinsic features of ACE could enhance tumor growth, mimicking the biological tumor microenvironment. Therefore, gastric cancer cells (AGS cell line) were cultured in contact with these matrices for up to 14 days to evaluate their cytocompatibility as a processed biomaterial. Results obtained by live-dead assay and MTS showed that cells are viable and metabolically active. Furthermore, confocal laser scanning microscopy analysis showed a homogeneous distribution of cells within the sponge. Therefore, the photocrosslinking on acemmann is proposed as an alternative approach that could significantly improve the processability of acemannan, allowing the production of high added-value structures enabling exploitation of its potential biomedical value. ; Portuguese Foundation for Science and Technology – FCT (SFRH/BPD/112140/2015, SFRH/BPD/85790/2012, respectively). This work was also financially supported by the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement no. REGPOT-CT2012-316331-POLARIS. The ...

Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by synovial joint inflammation and cartilage and bone tissue destruction. Although there exist some treatment strategies for RA, they are not completely safe and effective. Therefore, it is important to develop and test new drugs for RA that specifically target inflamed/swollen joints and simultaneously attenuate other possible damages to healthy tissues. Nanotechnology can be a good alternative to consider when envisioning precise medication for treating RA. Through the use of nanoparticles, it is possible to increase bioavailability and bioactivity of therapeutics and enable selective targeting to damaged joints. Herein, recent studies using nanoparticles for the treatment of RA, namely with liposomes, polymeric nanoparticles, dendrimers, and metallic nanoparticles, have been reviewed. These therapeutic strategies have shown great promise in improving the treatment over that by traditional drugs. The results of these studies confirm that feasibility of the use of nanoparticles is mainly due to their biocompatibility, low toxicity, controlled release, and selective drug delivery to inflamed tissues in animal RA models. Therefore, it is possible to claim that nanotechnology will, in the near future, play a crucial role in advanced treatments and patient-specific therapies for human diseases such as RA. ; Financial support under the ARTICULATE project (No. QREN-13/SI/2011-23189). This study was also funded by the Portuguese Foundation for Science and Technology (FCT) project OsteoCart (No. PTDC/CTM-BPC/115977/2009), as well as the European Union's FP7 Programme under grant agreement no REGPOT-CT2012-316331-POLARIS. The FCT distinction attributed to J. M. O. under the Investigator FCT program (No. IF/00423/2012) is also greatly acknowledged. C. G. also wished to acknowledge FCT for supporting her research (No. SFRH/BPD/94277/2013) ...

Functionalized electrospun fibers are of great interest for biomedical applications such as in the design of drug delivery systems. Nevertheless, in some cases the molecules of interest have poor solubility in water or have high melting temperatures. These drawbacks can be overcome using deep eutectic solvents. In this work, poly(vinyl alcohol) (PVA), a common biodegradable biopolymer, was used to produce new functionalized fibers with the eutectic mixture choline chloride:citric acid in a molar ratio of (1:1) ChCl:CA (1:1), which was used as a model system. Fibers were produced from an aqueous solution with 7.8% (w/v) and 9.8% (w/v) of 95% hydrolyzed PVA and a 2% (v/v) of ChCl:CA (1:1). Smooth, uniform fibers with an average diameter of 0.4 μm were obtained with a content of 19.8 wt % of ChCl:CA (1:1) encapsulated ; FCT/MEC through the project ENIGMA - PTDC/EQU-EPR/121491/2010, and the project PEst-C/ EQB/LA0006/2013. European Union's Seventh Framework Programme (FP7/2007-2013) under grant agreement n°REGPOT-CT2012-316331-POLARIS, and from the Project "Novel smart andbiomimetic materials for innovative regenerative medicine approaches (Ref.: RL1 - ABMR - NORTE-01-0124-FEDER-000016) "cofinanced by the North Portugal Regional Operational Programme (ON.2− O Novo Norte), under the National Strategic Reference Framework (NSRF), through the European Regional Development Fund (ERDF). This work was also partly funded by FEDER funds through the COMPETE 2020 Programme and National Funds through FCT - Portuguese Foundation for Science and Technology under the project ...

uncorrected proof ; This paper presents a novel approach towards the production of hybrid alginate-lignin aerogels. The key idea of the approach is to employ pressurized carbon dioxide for gelation. Exposure of alginate and lignin solution containing calcium carbonate to CO2 at 4.5 MPa resulted in a hydrogel formation. Various lignin and CaCO3 concentrations were studied. Stable hydrogels can be formed up to 2:1 (w/w) alginate-to-lignin ratio. Upon substitution of water with ethanol, gels were dried in supercritical CO2 to produce aerogels. Aerogels were characterized in terms of bulk density, specific surface area and pore volume. To introduce macroporosity, the CO2 induced gelation was supplemented with rapid depressurization (foaming process). Porosity, mean pore size and interconnectivity in the macroporous range were assessed by micro-CT. Youngâ s modulus of alginate-lignin aerogels was measured in both dry and wet state. Cell studies revealed that alginate-lignin aerogels are non-cytotoxic and demonstrate good cell adhesion properties making them attractive candidates for a wide range of applications including tissue engineering and regenerative medicine. ; Fundação da Ciência e Tecnologia (FCT) through the project ENIGMA – PTDC/EQU-EPR/121491/2010, and from the European Union's Seventh Framework Programme (FP7/2007-2013) under grant agreement no. REGPOT-CT2012-316331-POLARIS and from Project "Novel smart and biomimetic materials for innovative regenerative medicine approaches (Ref.: RL1 – ABMR – NORTE-01-0124-FEDER-000016)" cofinanced by North Portugal Regional Operational Programme (ON.2-O Novo Norte), under the National Strategic Reference Framework (NSRF), through the European Regional Development Fund (ERDF) and FEDER. Fundação da Ciência e Tecnologia (FCT) through the grant BIM/PTDC/EQU-EPR/121491/2010/ENIGMA, bilateral cooperation project FCT-DAAD 57036335, and from DFG (projects SM 82/8-1 and SM ...

The osteochondral tissue represents a complex structure composed of four interconnected structures, namely hyaline cartilage, a thin layer of calcified cartilage, subchondral bone, and cancellous bone. Due to the several difficulties associated with its repair and regeneration, researchers have developed several studies aiming to restore the native tissue, some of which had led to tissue-engineered commercial products. In this sense, this chapter discusses the good manufacturing practices, regulatory medical conditions and challenges on clinical translations that should be fulfilled regarding the safety and efficacy of the new commercialized products. Furthermore, we review the current osteochondral products that are currently being marketed and applied in the clinical setting, emphasizing the advantages and difficulties of each one. ; FROnTHERA (NORTE-01-0145- FEDER-000023), supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). The authors would also like to acknowledge H2020-MSCA-RISE program, as this work is part of developments carried out in BAMOS project, funded by the European Union's Horizon 2020 research and innovation program under grant agreement N° 734156. The financial support from the Portuguese Foundation for Science and Technology under the program Investigador FCT 2012 and 2015 (IF/00423/2012 and IF/01285/2015) ...

Juglans regia L. (walnut) leaves are considered a source of healthcare compounds, and have been widely used in traditional medicine. Furthermore, they have been reported as a source of bioactive molecules such as the antioxidant phenolic compounds. In the present study, organic acids, mono and oligosaccharides, and tocopherols were analyzed in leaves and decoction (widely used for topical applications). Furthermore, the phenolic composition and bioactivity of walnut leaves was determined in its decoction and methanol extract. Malic acid, sucrose, alpha-tocopherol and 3-O-caffeoylquinic acids and quercetin O-pentoside were the most abundant organic acid, disaccharide, tocopherol isomer and phenolic compounds, respectively. Methanol extract presented higher antioxidant and antitumor potential than decoction; both samples did not show toxicity for non-tumor liver primary cells. This study provides a more complete characterization of phenolic compounds of walnut leaves, and as far as we know, this is the first time that the presence of procyanidins and taxifolin derivatives, as also tocopherols were reported in J. regia leaves. ; The authors are grateful to Fundacao para a Ciencia e a Tecnologia (FCT, Portugal) for financial support to CIMO (strategic project PEst-OE/AGR/UI0690/2011). L. Barros and R.C. Calhelha also thank to FCT, POPH-QREN and FSE for their grants (SFRH/BPD/4609/2008 and SFRH/BPD/68344/2010, respectively). The GIP-USAL is financially supported by the Spanish Government through the Consolider-Ingenio 2010 Programme (FUN-C-FOOD, CSD2007-00063). M. Duenas thanks the Programa Ramon y Cajal for a ...

The musculoskeletal tissues have a prime role in the biomechanical support and metabolic activities of the human body. As musculoskeletal tissues are highly prone to injuries, conditions afflicting these tissues have a great impact on the quality of life of patients worldwide. Tissue engineering approaches hold the promise to develop bioengineered substitutes aiming at the regeneration of failing and injured tissue and organs. To effectively address the tissue-specific structural and biochemical features of musculoskeletal tissues, different biomaterials and techniques have been employed envisioning biomimetic solutions. Herein, the unique composition, structure, and function of the musculoskeletal tissues, namely bone, cartilage, and tendon, as well as state-of-the-art technologies to develop bioinspired strategies for tissue regeneration will be overviewed. Finally, this chapter will also discuss the unmet challenges and future perspectives in the field. ; FCT Project MagTT PTDC/CTM-CTM/29930/2017 (POCI-01- 0145-FEDER-29930) for A.I.G postdoc grant, the FCT Project PTDC/NAN-MAT/30595/2017 (POCI-01-0145-FEDER-30595) for P.S.B. postdoc grant, and for the assistant researcher contract (RL1) of M.T.R from the project "Accelerating tissue engineering and personalized medicine discoveries by the integration of key enabling nanotechnologies, marine-derived biomaterials and stem cells" supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). Authors acknowledge the financial support from the European Union Framework Programme for Research and Innovation HORIZON 2020, under the TEAMING Grant agreement No. 739572—The Discoveries CTR and the European Research Council 2017-CoG MagTendon (No. ...

Folic Acid (FA)-tagged protein nanoemulsions were found to be preferentially internalized on B-cell lymphoma cell line (A20 cell line), which, for the first time, are reported to express folate receptor (FR)-alpha. Carbon monoxide releasing molecule-2 (CORM-2) was incorporated in the oil phase of the initial formulation. FA-functionalized nanoemulsions loaded with CORM-2 exhibited a considerable antitumor effect and an increased survival of BALB/c mice bearing subcutaneous A20 lymphoma tumors. The developed nanoemulsions also demonstrated to be well tolerated by these immunocompetent mice. Thus, the results obtained in this study demonstrate that FA-tagged protein nanoemulsions can be successfully used in cancer therapy, with the important ability to delivery drugs intracellularly. ; SFRH/BD/81479/2011 and SFRH/BD/81269/2011 scholarships from Fundação para a Ciência e a Tecnologia (FCT). This work has received funding from the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement NMP4-LA-2009-228827 NANOFOL. This work was supported by FEDER through POFC-COMPETE and by Portuguese funds from FCT through the project ...

Gardnerella vaginalis is the most common species found in bacterial vaginosis (BV). However, it is also present in a significant proportion of healthy women and G. vaginalis vaginal colonization does not always lead to BV. In an effort to better understand the differences between G. vaginalis isolated from women with a positive (BV) versus a negative (non-BV) diagnosis of BV, we compared the virulence potential of 7 BV and 7 non-BV G. vaginalis isolates and assessed the virulence factors related to biofilm formation, namely: initial adhesion and cytotoxic effect, biofilm accumulation, susceptibility to antibiotics, and transcript levels of the known vaginolysin, and sialidase genes. Furthermore, we also determined the ability of G. vaginalis to displace lactobacilli previously adhered to HeLa cells. Our results showed that non-BV strains were less virulent than BV strains, as suggested by the lower cytotoxicity and initial adhesion to Hela cells. Significant differences in expression of known virulence genes were also detected, further suggesting a higher virulence potential of the BV associated G. vaginalis. Importantly, we demonstrated that BV associated G. vaginalis were able to displace pre-coated vaginal protective lactobacilli and we hypothesize this to be a trigger for BV development. ; European Union funds (FEDER/COMPETE) and by national funds (FCT) under the project with reference FCOMP-01-0124-FEDER-008991 (PTDC/BIA-MIC/098228/2008). FCT Strategic Project of UID/BIO/04469/2013 unit the project NORTE-07-0124-FEDER-000027, co-funded by the Programa Operacional Regional do Norte(ON.2 – O Novo Norte), QREN, FEDER, and the project RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462). FCT individual fellowship ...