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Claudia Botelho finished her PhD in Science Engineering by Porto University in 2005. From 2001 to 2006 most of her research was carried out at: Queen Mary London University.; Cambridge University, UK and Nara Institute of Science and Technology, Japan. After finishing her PhD she started working on the modulation of calcium phosphate ceramics aiming to developed custom-made ceramic prostheses for reconstructive surgery. A project that resulted in a worldwide patent. She continues to work on the development of strategies for personalised therapies, as each patient has its unique features. Focusing on the last 5 year period (2012-2019 – note that due to 2 maternity leaves). She published 2 scientific books, is edit 1 scientific book, 5 book chapters. She published 38 scientific papers, being 21 between 2012 and 2020. She was invited to give several talks (8; 4 in 2012/2019). Her work was presented over 40 times in conferences (14 oral presentations in 2012/2019 and 6 poster presentations in this period). In 2013-2015; 2017-2020 she was invited to lecturer at Minho University and also at the Instituto Politécnico de Viana. Her work was distinguished with several international prizes, being the most important the European Award on Biomaterials and Tissue Engineering Doctoral Award.
The main goal of this study was to chemically characterize an aqueous S. nigra flower extract and validate it as a bioactive agent. The elderflower aqueous extraction was performed at different temperatures (50, 70 and 90 °C). The extract obtained at 90 °C exhibited the highest phenolic content and antiradical activity. Therefore, this extract was analyzed by GC-MS and HPLC-MS, which allowed the identification of 46 compounds, being quercetin and chlorogenic acid derivatives representative of 86% of the total of phenolic compounds identified in hydrophilic fraction of the aqueous extract. Naringenin (27.2%) was the major compound present in the lipophilic fraction. The antiproliferative effects of the S. nigra extract were evaluated using the colon cancer cell lines RKO, HCT-116, Caco-2 and the extract’s antigenotoxic potential was evaluated by the Comet assay in RKO cells. The RKO cells were the most susceptible to S. nigra flower extract (IC50 = 1250 µg mL−1). Moreover, the extract showed antimicrobial activity against Gram-positive bacteria, particularly Staphylococcus aureus and S. epidermidis. These results show that S. nigra-based extracts can be an important dietary source of bioactive phenolic compounds that contribute to health-span improving life quality, demonstrating their potential as nutraceutical, functional foods and/or cosmetic components for therapeutic purposes.
Pedro Ferreira-Santos; Helder Badim; Ângelo C. Salvador; Armando J. D. Silvestre; Sónia A. O. Santos; Sílvia M. Rocha; Ana M. Sousa; Maria Olívia Pereira; Cristina Pereira Wilson; Cristina M. R. Rocha; José António Teixeira; Cláudia M. Botelho. Chemical Characterization of Sambucus nigra L. Flowers Aqueous Extract and Its Biological Implications. Biomolecules 2021, 11, 1222 .
AMA StylePedro Ferreira-Santos, Helder Badim, Ângelo C. Salvador, Armando J. D. Silvestre, Sónia A. O. Santos, Sílvia M. Rocha, Ana M. Sousa, Maria Olívia Pereira, Cristina Pereira Wilson, Cristina M. R. Rocha, José António Teixeira, Cláudia M. Botelho. Chemical Characterization of Sambucus nigra L. Flowers Aqueous Extract and Its Biological Implications. Biomolecules. 2021; 11 (8):1222.
Chicago/Turabian StylePedro Ferreira-Santos; Helder Badim; Ângelo C. Salvador; Armando J. D. Silvestre; Sónia A. O. Santos; Sílvia M. Rocha; Ana M. Sousa; Maria Olívia Pereira; Cristina Pereira Wilson; Cristina M. R. Rocha; José António Teixeira; Cláudia M. Botelho. 2021. "Chemical Characterization of Sambucus nigra L. Flowers Aqueous Extract and Its Biological Implications." Biomolecules 11, no. 8: 1222.
FAO estimates that in 2030 the poultry meat production could reach 120 million tons, which is a challenge in terms of waste management. Feathers are mainly composed of keratin, an important biomaterial. Using feathers as a source of keratin will minimize the waste generated, while contributing to supply an important material for several industries, such as pharmaceutical and biomedical. The peptides were extracted from the feathers by microbial degradation. In this study, we evaluated the peptides effect on keratinocyte metabolic activity and migration. The influence of these peptides on non-activated and activated macrophages was also assessed. It was demonstrated that depending on the keratin peptide fraction in contact with keratinocytes, it is possible to modulate the migration rate of the keratinocytes. Peptide fraction with low molecular weight increases migration, while peptides with a high range of molecular sizes decreases it. Some peptide fractions induce the secretion of TNF-α in non-activated macrophages and not on activated macrophages, demonstrating that these peptides should only be placed in contact with cells, in the context of an ongoing inflammatory process. This work is a step forward on the understanding of keratin peptides influence on keratinocytes and immune cells system cells, macrophages.
Cláudia Botelho; Pedro Ferreira-Santos; Duarte Toubarro; Hugo Dinis; Hugo Osório; Augusto Costa-Barbosa; Paula Sampaio; Nelson Simões; José Teixeira. Chicken Feather Keratin Peptides for the Control of Keratinocyte Migration. Applied Sciences 2021, 11, 6779 .
AMA StyleCláudia Botelho, Pedro Ferreira-Santos, Duarte Toubarro, Hugo Dinis, Hugo Osório, Augusto Costa-Barbosa, Paula Sampaio, Nelson Simões, José Teixeira. Chicken Feather Keratin Peptides for the Control of Keratinocyte Migration. Applied Sciences. 2021; 11 (15):6779.
Chicago/Turabian StyleCláudia Botelho; Pedro Ferreira-Santos; Duarte Toubarro; Hugo Dinis; Hugo Osório; Augusto Costa-Barbosa; Paula Sampaio; Nelson Simões; José Teixeira. 2021. "Chicken Feather Keratin Peptides for the Control of Keratinocyte Migration." Applied Sciences 11, no. 15: 6779.
Polyphenolic extracts from pine bark have reported different biological actions and promising beneficial effects on human health. However, its susceptibility to environmental stresses (temperature, storage, etc.) and physiological human conditions prequires the development of efficient protection mechanisms to allow effective delivering of functionality. The aim of this work was to encapsulate pine bark extract rich phenolic compounds by spray-drying using maltodextrin, and understand the influence of encapsulation on the antioxidant and antimicrobial activity and bioaccessibility of phenolic compounds during gastrointestinal digestion. The optimized process conditions allowed good encapsulation efficiency of antioxidant phenolic compounds. The microencapsulation was effective in protecting those compounds during gastrointestinal conditions, controlling their delivery and enhancing its health benefits, decreasing the production of reactive oxygen species implicated in the process of oxidative stress associated with some pathologies. Finally, this encapsulation system was able to protect these extracts against acidic matrices, making the system suitable for the nutritional enrichment of fermented foods or fruit-based beverages, providing them antimicrobial protection, because the encapsulated extract was effective against Listeria innocua. Overall, the designed system allowed protecting and appropriately delivering the active compounds, and may find potential application as a natural preservative and/or antioxidant in food formulations or as bioactive ingredient with controlled delivery in pharmaceuticals or nutraceuticals.
Pedro Ferreira-Santos; Raquel Ibarz; Jean-Michel Fernandes; Ana Pinheiro; Cláudia Botelho; Cristina Rocha; José Teixeira; Olga Martín-Belloso. Encapsulated Pine Bark Polyphenolic Extract during Gastrointestinal Digestion: Bioaccessibility, Bioactivity and Oxidative Stress Prevention. Foods 2021, 10, 328 .
AMA StylePedro Ferreira-Santos, Raquel Ibarz, Jean-Michel Fernandes, Ana Pinheiro, Cláudia Botelho, Cristina Rocha, José Teixeira, Olga Martín-Belloso. Encapsulated Pine Bark Polyphenolic Extract during Gastrointestinal Digestion: Bioaccessibility, Bioactivity and Oxidative Stress Prevention. Foods. 2021; 10 (2):328.
Chicago/Turabian StylePedro Ferreira-Santos; Raquel Ibarz; Jean-Michel Fernandes; Ana Pinheiro; Cláudia Botelho; Cristina Rocha; José Teixeira; Olga Martín-Belloso. 2021. "Encapsulated Pine Bark Polyphenolic Extract during Gastrointestinal Digestion: Bioaccessibility, Bioactivity and Oxidative Stress Prevention." Foods 10, no. 2: 328.
Fabric structures are prone to contamination with microorganisms, as their morphology and ability to retain moisture creates a proper environment for their growth. In this work, a novel, easily processed and cheap coating for a nylon fabric with antimicrobial characteristics was developed. After plasma treatment, made to render the fabric surface more reactive sites, the fabric was impregnated with chitosan and silver nanoparticles by simply dipping it into a mixture of different concentrations of both components. Silver nanoparticles were previously synthesized using the Lee–Meisel method, and their successful obtention was proven by UV–Vis, showing the presence of the surface plasmon resonance band at 410 nm. Nanoparticles with 25 nm average diameter observed by STEM were stable, mainly in the presence of chitosan, which acted as a surfactant for silver nanoparticles, avoiding their aggregation. The impregnated fabric possessed bactericidal activity higher for Gram-positive Staphylococcus aureus than for Gram-negative Pseudomonas aeruginosa bacteria for all combinations. The percentage of live S. aureus and P. aeruginosa CFU was reduced to less than 20% and 60%, respectively, when exposed to each of the coating combinations. The effect was more pronounced when both chitosan and silver were present in the coating, suggesting an effective synergy between these components. After a washing process, the antimicrobial effect was highly reduced, suggesting that the coating is unstable after washing, being almost completely removed from the fabric. Nevertheless, the new-coated fabric can be successfully used in single-use face masks. To our knowledge, the coating of nylon fabrics intended for face-mask material with both agents has never been reported.
Cláudia M. Botelho; Margarida M. Fernandes; Jefferson M. Souza; Nicolina Dias; Ana M. Sousa; José A. Teixeira; Raul Fangueiro; Andrea Zille. New Textile for Personal Protective Equipment—Plasma Chitosan/Silver Nanoparticles Nylon Fabric. Fibers 2021, 9, 3 .
AMA StyleCláudia M. Botelho, Margarida M. Fernandes, Jefferson M. Souza, Nicolina Dias, Ana M. Sousa, José A. Teixeira, Raul Fangueiro, Andrea Zille. New Textile for Personal Protective Equipment—Plasma Chitosan/Silver Nanoparticles Nylon Fabric. Fibers. 2021; 9 (1):3.
Chicago/Turabian StyleCláudia M. Botelho; Margarida M. Fernandes; Jefferson M. Souza; Nicolina Dias; Ana M. Sousa; José A. Teixeira; Raul Fangueiro; Andrea Zille. 2021. "New Textile for Personal Protective Equipment—Plasma Chitosan/Silver Nanoparticles Nylon Fabric." Fibers 9, no. 1: 3.
Non-viral gene therapy based on gene silencing with small interfering RNA (siRNA) has attracted great interest over recent years. Among various types of cationic complexation agents, amino acid-based surfactants have been recently explored for nucleic acid delivery due to their low toxicity and high biocompatibility. Monoolein (MO), in turn, has been used as helper lipid in liposomal systems due to its ability to form inverted nonbilayer structures that enhance fusogenicity, thus contributing to higher transfection efficiency. In this work, we focused on the development of nanovectors for siRNA delivery based on three gemini amino acid-based surfactants derived from serine — (12Ser)2N12, amine derivative; (12Ser)2COO12, ester derivative; and (12Ser)2CON12, amide derivative — individually combined with MO as helper lipid. The inclusion of MO in the cationic surfactant system influences the morphology and size of the mixed aggregates. Furthermore, the gemini surfactant:MO systems showed the ability to efficiently complex siRNA, forming stable lipoplexes, in some cases clearly depending on the MO content, without inducing significant levels of cytotoxicity. High levels of gene silencing were achieved in comparison with a commercially available standard indicating that these gemini:MO systems are promising candidates as lipofection vectors for RNA interference (RNAi)-based therapies.
Catarina Costa; Isabel S. Oliveira; João P. N. Silva; Sandra G. Silva; Cláudia Botelho; M. Luísa C. Do Vale; Maria Elisabete C. D. Real Oliveira; Andreia C. Gomes; Eduardo F. Marques. Effective cytocompatible nanovectors based on serine-derived gemini surfactants and monoolein for small interfering RNA delivery. Journal of Colloid and Interface Science 2020, 584, 34 -44.
AMA StyleCatarina Costa, Isabel S. Oliveira, João P. N. Silva, Sandra G. Silva, Cláudia Botelho, M. Luísa C. Do Vale, Maria Elisabete C. D. Real Oliveira, Andreia C. Gomes, Eduardo F. Marques. Effective cytocompatible nanovectors based on serine-derived gemini surfactants and monoolein for small interfering RNA delivery. Journal of Colloid and Interface Science. 2020; 584 ():34-44.
Chicago/Turabian StyleCatarina Costa; Isabel S. Oliveira; João P. N. Silva; Sandra G. Silva; Cláudia Botelho; M. Luísa C. Do Vale; Maria Elisabete C. D. Real Oliveira; Andreia C. Gomes; Eduardo F. Marques. 2020. "Effective cytocompatible nanovectors based on serine-derived gemini surfactants and monoolein for small interfering RNA delivery." Journal of Colloid and Interface Science 584, no. : 34-44.
Nanoparticles are one of the most commonly used systems for imaging or therapeutic drug delivery. Exosomes are nanovesicular carriers that transport cargo for intercellular communication. These nanovesicles are linked to the pathology of some major diseases, in some cases with a central role in their progression. The use of these carriers to transport therapeutic drugs is a recent and promising approach to treat diseases such as cancer and Alzheimer disease. The physiological production of these structures is limited impairing its collection and subsequent purification. These drawbacks inspired the search for mimetic alternatives. The collection of exosome-like nanoparticles from plants can be a good alternative, since they are easier to extract and do not have the drawbacks of those produced in animal cells. Both natural and synthetic exosome-like nanoparticles, produced from serial extrusion of cells or by bottom up synthesis, are currently some of the most promising, biocompatible, high efficiency systems for drug delivery.
Mário Fernandes; Ivo Lopes; José Teixeira; Cláudia Botelho; Andreia Gomes. Exosome-like Nanoparticles: A New Type of Nanocarrier. Current Medicinal Chemistry 2020, 27, 3888 -3905.
AMA StyleMário Fernandes, Ivo Lopes, José Teixeira, Cláudia Botelho, Andreia Gomes. Exosome-like Nanoparticles: A New Type of Nanocarrier. Current Medicinal Chemistry. 2020; 27 (23):3888-3905.
Chicago/Turabian StyleMário Fernandes; Ivo Lopes; José Teixeira; Cláudia Botelho; Andreia Gomes. 2020. "Exosome-like Nanoparticles: A New Type of Nanocarrier." Current Medicinal Chemistry 27, no. 23: 3888-3905.
Natural compounds from agro-food by-products have fostered interest in food industries. The aim of this study was to unravel potential uses for Pinus pinaster bark extracts (PBE). As functional features of this type of extracts are usually attributed to phenolic compounds, the extraction process was studied. Different PBEs were achieved, with high content in phenolic compounds, using different water/ethanol combinations as a solvent. These PBEs were chemically characterized, and their bioactivity and in vitro cell viability were evaluated. Extracts obtained with hydroethanolic solvents had higher content in phenolic and flavonoid compounds. All the PBEs presented high antioxidant, antibacterial and antihyperglycemic activities. Moreover, PBEs have low cytotoxicity and a selective activity against cancer cells as these were negatively affected. These features may allow the extracts to be used in food formulation and processing (as preservatives, antioxidants or bioactive ingredients), but they showed also potential for the pharmaceutical or nutraceutical sectors.
Pedro Miguel Ferreira Santos; Zlatina Genisheva; Cláudia Botelho; Joana Santos; Carla Ramos; José A. Teixeira; Cristina M.R. Rocha. Unravelling the Biological Potential of Pinus pinaster Bark Extracts. Antioxidants 2020, 9, 334 .
AMA StylePedro Miguel Ferreira Santos, Zlatina Genisheva, Cláudia Botelho, Joana Santos, Carla Ramos, José A. Teixeira, Cristina M.R. Rocha. Unravelling the Biological Potential of Pinus pinaster Bark Extracts. Antioxidants. 2020; 9 (4):334.
Chicago/Turabian StylePedro Miguel Ferreira Santos; Zlatina Genisheva; Cláudia Botelho; Joana Santos; Carla Ramos; José A. Teixeira; Cristina M.R. Rocha. 2020. "Unravelling the Biological Potential of Pinus pinaster Bark Extracts." Antioxidants 9, no. 4: 334.
Epidermal Growth Factor Receptor (EGFR) plays a key role in regulating cell survival, proliferation and migration, and its overexpression and activation has been correlated with cancer progression. Cancer therapies targeting EGFR have been applied in the clinic with some success. We show, by confocal microscopy analysis, that illumination of adenocarcinomic human alveolar basal epithelial cells (Human A549 – EGFR biosensor cell line) with 280 nm at irradiance levels up to 20 times weaker than the UVB solar output for short periods of time (15-45 min) prevents EGF-mediated activation of EGFR located on the cell membrane, preventing or reducing cellular disaggregation, formation of filopodia and cell migration. This effect of UV light illumination was confirmed further in a functional scratch assay, and shown to be more effective than that of a specific EGFR-signaling inhibitor. This new photonic approach may be applicable to the treatment of various types of cancer, alone or in combination with other therapies.
Cláudia Manuela Botelho; Odete Gonçalves; Rogerio Marques; Viruthachalam Thiagarajan; Henrik Vorum; Andreia Castro Gomes; Maria Teresa Neves-Petersen. Photonic modulation of epidermal growth factor receptor halts receptor activation and cancer cell migration. Journal of Biophotonics 2018, 11, e201700323 .
AMA StyleCláudia Manuela Botelho, Odete Gonçalves, Rogerio Marques, Viruthachalam Thiagarajan, Henrik Vorum, Andreia Castro Gomes, Maria Teresa Neves-Petersen. Photonic modulation of epidermal growth factor receptor halts receptor activation and cancer cell migration. Journal of Biophotonics. 2018; 11 (9):e201700323.
Chicago/Turabian StyleCláudia Manuela Botelho; Odete Gonçalves; Rogerio Marques; Viruthachalam Thiagarajan; Henrik Vorum; Andreia Castro Gomes; Maria Teresa Neves-Petersen. 2018. "Photonic modulation of epidermal growth factor receptor halts receptor activation and cancer cell migration." Journal of Biophotonics 11, no. 9: e201700323.
Diospyros kaki (Dk) leaves have been used in China, Korea and Japan as a delicate, pleasant beverage. Additionally, it is used as a folk remedy for hypertension, ischemic stroke, angina and internal hemorrhage. However, in EU and particularly in Portugal Dk leaves are considered an agriculture waste. The aim of this study was to characterize methanolic Dk leaf extract concerning its in vitro antioxidant proprieties and evaluate eventual cytotoxic effects, using HepG2 cells (hepatocytes). HPLC-DAD profiles of Dk leaves showed that extracts are mainly composed by phenolic compounds (flavonoids and phenolic acids derivatives), known antioxidants. To assess antioxidant potential, we used several in vitro tests including DPPH scavenging, iron chelation, inhibition of nitric oxide and superoxide scavenging. Results showed that Dk have a high capacity to scavenge superoxide radicals (EC50 8.94 µg/ml), even better than quercetin (EC50 18.73 µg/ml), and DPPH radicals (EC50 15.27 µg/ml). For iron chelating activity and inhibition of nitric oxide production Dk extract had lower potential with and EC50 of 1376 and 1027 µg/ml, respectively. So, we can conclude that Dk leaf extract is a good radical scavenger and might protect cells from oxidative stress. Since Dk leaves are used in beverages and liver is always a key target organ for metabolization and detoxification, we tested possible cytotoxicity on HepG2 cells. Results did not show significant cytotoxic effects of Dk extracts on HepG2 cells (using MTS evaluation), up to 1 mg/ml, for 3 and 24h incubation times. Hence, we can infer that Dk leaf extracts have good antioxidant potential and no significant toxicity (at least on hepatocytes), however more tests are required with other cellular models. This work was supported by project HEALTHKAKI, N° 010672, Programa Operacional Regional do Norte, no âmbito do Portugal 2020.
V Castro; A Gomes; C Botelho; A Dias. Evaluation of antioxidant potential and cytotoxic effects of alcoholic extracts of Diospyros kaki leaves. 65th International Congress and Annual Meeting of the Society for Medicinal Plant and Natural Product Research (GA 2017) 2017, 4, Mo-PO-138 .
AMA StyleV Castro, A Gomes, C Botelho, A Dias. Evaluation of antioxidant potential and cytotoxic effects of alcoholic extracts of Diospyros kaki leaves. 65th International Congress and Annual Meeting of the Society for Medicinal Plant and Natural Product Research (GA 2017). 2017; 4 (S 01):Mo-PO-138.
Chicago/Turabian StyleV Castro; A Gomes; C Botelho; A Dias. 2017. "Evaluation of antioxidant potential and cytotoxic effects of alcoholic extracts of Diospyros kaki leaves." 65th International Congress and Annual Meeting of the Society for Medicinal Plant and Natural Product Research (GA 2017) 4, no. S 01: Mo-PO-138.
Overexpression of the Epidermal Growth Factor Receptor (EGFR) by cancer cells is associated with a poor prognosis for the patient. For several decades, therapies targeting EGFR have been designed, including the use of monoclonal antibodies and small molecule tyrosine kinase inhibitors. The use of these molecules had good clinical results, although its efficiency (and specificity) is still far from being optimal. In this paper, we present a new approach for a possible new cancer therapy targeting EGFR and using low intensity 280nm light. The influence of 280nm UVB illumination on cancer cells stimulated with 2nM of EGF was followed by time-lapse confocal microscopy. The 280nm illumination of the cancer cells blocks EGFR activation, inhibiting EGFR internalization and cell migration thus inhibiting the transition to the metastatic phenotype. Exposure time is a very important factor. The higher the illumination time the more significant differences were observed: 280nm light delayed or completely halted EGFR activation in the cell membrane, mainly at the cell junction level, and delayed or halted EGFR endocytic internalization, filopodia formation and cell migration. © (2017) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Cláudia M. Botelho; Rogério Marques; Thiagarajan Viruthachalam; Odete Gonçalves; Henrik Vorum; Andreia C. Gomes; Maria Teresa Neves-Petersen. Photonic modulation of EGFR: 280nm low level light arrests cancer cell activation and migration. Mechanisms of Photobiomodulation Therapy XII 2017, 10048, 1 .
AMA StyleCláudia M. Botelho, Rogério Marques, Thiagarajan Viruthachalam, Odete Gonçalves, Henrik Vorum, Andreia C. Gomes, Maria Teresa Neves-Petersen. Photonic modulation of EGFR: 280nm low level light arrests cancer cell activation and migration. Mechanisms of Photobiomodulation Therapy XII. 2017; 10048 ():1.
Chicago/Turabian StyleCláudia M. Botelho; Rogério Marques; Thiagarajan Viruthachalam; Odete Gonçalves; Henrik Vorum; Andreia C. Gomes; Maria Teresa Neves-Petersen. 2017. "Photonic modulation of EGFR: 280nm low level light arrests cancer cell activation and migration." Mechanisms of Photobiomodulation Therapy XII 10048, no. : 1.
Biofilms plays an important role in medical device-related infections. This study aimed to determine the factors that influence adherence and biofilm production, as well as the relationship between strong biofilm production and genetic determinants in clinical isolates of MRSA. Fifteen strains carrying different chromosomal cassettes, recovered from patients hospitalized were selected: five SCCmecII, five SCCmecIII and five SCCmecIV. The SCCmec type, agr group and the presence of the virulence genes (bbp, clfA, icaA, icaD, fnbB, bap, sasC and IS256) were assessed by PCR. Pulsed field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) techniques also were performed. The initial adhesion and biofilm formation were examined by quantitative assays. The surface tension and hydrophobicity of the strains were measured by contact angle technique to evaluate the association between these parameters and adhesion ability. SCCmecIII and IV strains were less hydrophilic, with a high value for the electron acceptor parameter and higher adhesion in comparison with SCCmecII strains. Only SCCmecIII strains could be characterized as strong biofilm producers. The PFGE showed five major pulsotypes (A-E) however, biofilm production was related to the dissemination of one specific PFGE clone (C) belonging to MLST ST239 (BECC, Brazilian epidemic clonal complex). The genes agrI, fnbB and IS256 in SCCmecIII strains, were considered as genetic determinants associated with strong biofilm-formation by an ica-independent biofilm pathway. This study contributes to the understanding of biofilm production as an aggravating factor potentially involved in the persistence and severity of infections caused by multidrug-resistant MRSA belonging to this genotype.
Deivid William Fonseca Batistão; Paola Amaral De Campos; Nayara Caroline Camilo; Sabrina Royer; Bruna Fuga Araújo; Karinne Spirandelli Carvalho Naves; Margarida Martins; Maria Olivia Pereira; Mariana Henriques; Paulo Pinto Gontijo-Filho; Cláudia Botelho; Rosário Oliveira; Rosineide Marques Ribas. Biofilm formation of Brazilian meticillin-resistant Staphylococcus aureus strains: prevalence of biofilm determinants and clonal profiles. Journal of Medical Microbiology 2016, 65, 286 -297.
AMA StyleDeivid William Fonseca Batistão, Paola Amaral De Campos, Nayara Caroline Camilo, Sabrina Royer, Bruna Fuga Araújo, Karinne Spirandelli Carvalho Naves, Margarida Martins, Maria Olivia Pereira, Mariana Henriques, Paulo Pinto Gontijo-Filho, Cláudia Botelho, Rosário Oliveira, Rosineide Marques Ribas. Biofilm formation of Brazilian meticillin-resistant Staphylococcus aureus strains: prevalence of biofilm determinants and clonal profiles. Journal of Medical Microbiology. 2016; 65 (4):286-297.
Chicago/Turabian StyleDeivid William Fonseca Batistão; Paola Amaral De Campos; Nayara Caroline Camilo; Sabrina Royer; Bruna Fuga Araújo; Karinne Spirandelli Carvalho Naves; Margarida Martins; Maria Olivia Pereira; Mariana Henriques; Paulo Pinto Gontijo-Filho; Cláudia Botelho; Rosário Oliveira; Rosineide Marques Ribas. 2016. "Biofilm formation of Brazilian meticillin-resistant Staphylococcus aureus strains: prevalence of biofilm determinants and clonal profiles." Journal of Medical Microbiology 65, no. 4: 286-297.
The synthesis of biodiesel from sunflower oil and methanol based on transesterification using the immobilized lipase from Thermomyces lanuginosus (Lipozyme TL-IM) has been investigated under silent conditions and under an ultrasound field. Ultrasound assisted process led to reduced processing time and requirement of lower enzyme dosage. We found for the first time that oil to methanol ratio of 1:3 was favored for the ultrasound assisted enzymatic process which is lower than that observed for the case of conventional stirring based approach (ratio of 1.4). Our results indicate that intensification provided by ultrasound suppresses the need of the excess of the methanol reactant during the enzymatic biodiesel production. Ultrasound assisted enzymatic biodiesel production is therefore a faster and a cleaner processes.
Preeti B. Subhedar; Claudia Botelho; Artur Ribeiro; Rita Castro; Maria Alcina Pereira; Parag R. Gogate; Artur Cavaco-Paulo. Ultrasound intensification suppresses the need of methanol excess during the biodiesel production with Lipozyme TL-IM. Ultrasonics Sonochemistry 2015, 27, 530 -535.
AMA StylePreeti B. Subhedar, Claudia Botelho, Artur Ribeiro, Rita Castro, Maria Alcina Pereira, Parag R. Gogate, Artur Cavaco-Paulo. Ultrasound intensification suppresses the need of methanol excess during the biodiesel production with Lipozyme TL-IM. Ultrasonics Sonochemistry. 2015; 27 ():530-535.
Chicago/Turabian StylePreeti B. Subhedar; Claudia Botelho; Artur Ribeiro; Rita Castro; Maria Alcina Pereira; Parag R. Gogate; Artur Cavaco-Paulo. 2015. "Ultrasound intensification suppresses the need of methanol excess during the biodiesel production with Lipozyme TL-IM." Ultrasonics Sonochemistry 27, no. : 530-535.
While the delivery of small interfering RNAs (siRNAs) is an attractive strategy to treat several clinical conditions, siRNA-nanocarriers’ stability after intravenous administration is still a major obstacle for the development of RNA-interference based therapies. But, although the need for stability is well recognized, the notion that strong stabilization can decrease nanocarriers’ efficiency is sometimes neglected. In this work we evaluated two stealth functionalization strategies to stabilize the previously validated dioctadecyldimethylammonium bromide (DODAB):monoolein (MO) siRNA-lipoplexes. The nanocarriers were pre- and post-pegylated, forming vectors with different stabilities in biological fluids. The stealth nanocarriers’ behavior was tested under biological mimetic conditions, as the production of stable siRNA-lipoplexes is determinant to achieve efficient intravenous siRNA delivery to cancer cells. Upon incubation in human serum for 2 h, by fluorescence Single Particle Tracking microscopy, PEG-coated lipoplexes were found to have better colloidal stability as they could maintain a relatively stable size. In addition, using fluorescence fluctuation spectroscopy, post-pegylation also proved to avoid siRNA dissociation from the nanocarriers in human serum. Concomitantly it was found that PEG-coated lipoplexes improved cellular uptake and transfection efficiency in H1299 cells, and had the ability to silence BCR-ABL, affecting the survival of K562 cells. Based on an efficient cellular internalization, good silencing effect, good siRNA retention and good colloidal stability in human serum, DODAB:MO (2:1) siRNA-lipoplexes coated with PEG-Cer are considered promising nanocarriers for further in vivo validation. Statement of Significance This work describes two stealth functionalization strategies for the stabilization of the previously validated dioctadecyldimethylammonium bromide (DODAB):monoolein (MO) siRNA-lipoplexes. These nanocarriers are capable of efficiently incorporating and delivering siRNA molecules to cells in order to silence genes whose expression is implicated in a pathological condition. The main objective was to functionalize these nanocarriers with a coating conferring protection to siRNA in blood without compromising its efficient delivery to cancer cells, validating the potential of DODAB:MO (2:1) siRNA-lipoplexes as therapeutic vectors. We show that the stealth strategy is determinant to achieve a stable and efficient nanocarrier, and that DODAB:MO mixtures have a very promising potential for systemic siRNA delivery to leukemic cells.
Ana Oliveira; Koen Raemdonck; Thomas Martens; Koen Rombouts; Rosana Simón-Vázquez; Cláudia Botelho; Ivo Lopes; Marlene Lúcio; África González-Fernández; M. Elisabete C.D. Real Oliveira; Andreia C. Gomes; Kevin Braeckmans. Stealth monoolein-based nanocarriers for delivery of siRNA to cancer cells. Acta Biomaterialia 2015, 25, 216 -229.
AMA StyleAna Oliveira, Koen Raemdonck, Thomas Martens, Koen Rombouts, Rosana Simón-Vázquez, Cláudia Botelho, Ivo Lopes, Marlene Lúcio, África González-Fernández, M. Elisabete C.D. Real Oliveira, Andreia C. Gomes, Kevin Braeckmans. Stealth monoolein-based nanocarriers for delivery of siRNA to cancer cells. Acta Biomaterialia. 2015; 25 ():216-229.
Chicago/Turabian StyleAna Oliveira; Koen Raemdonck; Thomas Martens; Koen Rombouts; Rosana Simón-Vázquez; Cláudia Botelho; Ivo Lopes; Marlene Lúcio; África González-Fernández; M. Elisabete C.D. Real Oliveira; Andreia C. Gomes; Kevin Braeckmans. 2015. "Stealth monoolein-based nanocarriers for delivery of siRNA to cancer cells." Acta Biomaterialia 25, no. : 216-229.
Candida bracarensis is an uncommon Candida species found during an epidemiological study of candidiasis performed in Braga, Portugal. Initially, it was identified as C. glabrata, but recently detailed analyses pointed out their differences. So, little information is still available about C. bracarensis virulence factors and antifungal susceptibilities. Therefore, the main goal of this work is to evaluate the ability of C. bracarensis to form biofilms, to produce hydrolytic enzymes (proteases, phospholipases and hemolysins), as well as its susceptibility to amphotericin B and fluconazole. It was shown, for the first time, that all C. bracarensis strains were able to form biofilms and display proteinase and hemolytic activities. Moreover, although planktonic cells presented antifungal susceptibility, amphotericin B and fluconazole were unable to inhibit biofilm formation and eradicate pre-formed biofilms. Due to the propensity of C. bracarensis to display antifungal resistance and virulence attributes, the control of these emerging pathogens is recommended.
André Moreira; Sónia Silva; Cláudia Botelho; Paula Sampaio; Célia Pais; Mariana Henriques. Candida bracarensis: Evaluation of Virulence Factors and its Tolerance to Amphotericin B and Fluconazole. Mycopathologia 2015, 180, 305 -315.
AMA StyleAndré Moreira, Sónia Silva, Cláudia Botelho, Paula Sampaio, Célia Pais, Mariana Henriques. Candida bracarensis: Evaluation of Virulence Factors and its Tolerance to Amphotericin B and Fluconazole. Mycopathologia. 2015; 180 (5-6):305-315.
Chicago/Turabian StyleAndré Moreira; Sónia Silva; Cláudia Botelho; Paula Sampaio; Célia Pais; Mariana Henriques. 2015. "Candida bracarensis: Evaluation of Virulence Factors and its Tolerance to Amphotericin B and Fluconazole." Mycopathologia 180, no. 5-6: 305-315.
Poly(hydroxybenzene)-trimethoprim conjugates were prepared using methylparaben as substrate of the oxida- tive enzyme tyrosinase. MALDI-TOF MS analysis showed that the enzymatic oxidation of methylparaben alone leads to the poly(hydroxybenzene) formation. In the presence of tri- methoprim, the methylparaben tyrosinase oxidation leads poly(hydroxybenzene)-trimethoprim conjugates. All of these compounds were incorporated into lubricant hydroxyethyl cellulose/glycerol mixtures. Poly(hydroxybenzene)-trimetho- prim conjugates were the most effective phenolic structures against the bacterial growth reducing by 96 and 97 % of Escherichia coli and Staphylococcus epidermidis suspen- sions, respectively (after 24 h). A novel enzymatic strategy to produce antimicrobial poly(hydroxybenzene)-antibiotic conjugates is proposed here for a wide range of applications on the biomedical field.The authors Idalina Gonçalves and Cláudia Botelho would like to acknowledge the NOVO project (FP7-HEALTH- 2011.2.3.1- 5) for funding. Loïc Hilliou acknowledges the financial support by FCT – Foundation for Science and Technology, Portugal (501100001871), through Grant PEst-C/CTM/LA0025/2013 - Strategic Project - LA 25 - 2013–2014, and by Programa Operacional Regional do Norte (ON.2) through the project BMatepro – Optimizing Materials and Processes^, with reference NORTE-07-0124-FEDER-000037 FEDER COMPETE
Idalina Gonçalves; Claudia Botelho; Ana Teixeira; Ana S. Abreu; Loic Hilliou; Carla Silva; Artur Cavaco-Paulo. Antimicrobial lubricant formulations containing poly(hydroxybenzene)-trimethoprim conjugates synthesized by tyrosinase. Applied Microbiology and Biotechnology 2015, 99, 4225 -4235.
AMA StyleIdalina Gonçalves, Claudia Botelho, Ana Teixeira, Ana S. Abreu, Loic Hilliou, Carla Silva, Artur Cavaco-Paulo. Antimicrobial lubricant formulations containing poly(hydroxybenzene)-trimethoprim conjugates synthesized by tyrosinase. Applied Microbiology and Biotechnology. 2015; 99 (10):4225-4235.
Chicago/Turabian StyleIdalina Gonçalves; Claudia Botelho; Ana Teixeira; Ana S. Abreu; Loic Hilliou; Carla Silva; Artur Cavaco-Paulo. 2015. "Antimicrobial lubricant formulations containing poly(hydroxybenzene)-trimethoprim conjugates synthesized by tyrosinase." Applied Microbiology and Biotechnology 99, no. 10: 4225-4235.
Ana V. Ferreira; Vadim Volkov; Ana S. Abreu; Nuno Azoia; Cláudia M. Botelho; Artur Cavaco-Paulo. Orange IV stabilizes silk fibroin microemulsions. Engineering in Life Sciences 2015, 15, 400 -409.
AMA StyleAna V. Ferreira, Vadim Volkov, Ana S. Abreu, Nuno Azoia, Cláudia M. Botelho, Artur Cavaco-Paulo. Orange IV stabilizes silk fibroin microemulsions. Engineering in Life Sciences. 2015; 15 (4):400-409.
Chicago/Turabian StyleAna V. Ferreira; Vadim Volkov; Ana S. Abreu; Nuno Azoia; Cláudia M. Botelho; Artur Cavaco-Paulo. 2015. "Orange IV stabilizes silk fibroin microemulsions." Engineering in Life Sciences 15, no. 4: 400-409.
Objectives: This research work intends to clarify the role of artificial saliva, in particularly the role of mucin, a salivary protein, on the surface properties and adhesion ability of Candida spp. oral clinical isolates to abiotic surfaces. Methods: Four oral clinical isolates of Candida spp. were used: two Candida albicans strains (AC; AM) and two Candida parapsilosis strains (AD; AM2). The strains were isolated from patients using oral prosthesis. The microorganisms were cultured in the absence or presence of mucin and artificial saliva, and their adhesion to an abiotic surface (coated with mucin and artificial saliva) was evaluated. Results: The presence of mucin per se onto the abiotic surface decreased the adhesion of all strains, although the combination of mucin with artificial saliva had reduced this effect. No direct correlation between adhesion and the surface free energies of adhesion of the microorganisms was found. Significance: Candida spp. were human commensal microorganisms that became pathogenic when the host immune defenses were compromised. Medical devices were colonized by Candida spp. particularly, oral prostheses, which might lead to the degradation of the prostheses and systemic infections. The salivary secretions that constantly cover the oral cavity influenced Candida spp. adhesion process. Therefore, it was important to understand the interactions between Candida spp., salivary proteins and the characteristic of oral prosthesis when developing materials for oral prostheses.
Catarina L. Seabra; Claudia Botelho; Ana C. N. Oliveira; Mariana Henriques. Influence of Saliva and Mucin on the Adhesion of Candida Oral Clinical Isolates. Journal of Encapsulation and Adsorption Sciences 2015, 05, 217 -227.
AMA StyleCatarina L. Seabra, Claudia Botelho, Ana C. N. Oliveira, Mariana Henriques. Influence of Saliva and Mucin on the Adhesion of Candida Oral Clinical Isolates. Journal of Encapsulation and Adsorption Sciences. 2015; 05 (04):217-227.
Chicago/Turabian StyleCatarina L. Seabra; Claudia Botelho; Ana C. N. Oliveira; Mariana Henriques. 2015. "Influence of Saliva and Mucin on the Adhesion of Candida Oral Clinical Isolates." Journal of Encapsulation and Adsorption Sciences 05, no. 04: 217-227.
We have previously demonstrated that the C. albicans pre-vacuolar protein sorting gene VPS4 is required for extracellular secretion of the secreted aspartyl proteases Sap2p and Saps4-6p. Furthermore, the vps4Δ null mutant has been shown to be markedly hypovirulent in a murine tail vein model of disseminated candidiasis. In these experiments, we sought to further define the role of the pre-vacuolar secretion pathway mediated by the pre-vacuolar sorting gene VPS4 in the pathogenesis of epithelial and mucosal infection using a broad range of virulence models. The C. albicans vps4Δ mutant demonstrates reduced tolerance of cell wall stresses compared to its isogenic, complemented control strain. In an in vitro oral epithelial model (OEM) of tissue invasion, the vps4Δ mutant caused reduced tissue damage compared to controls. Further, the vps4Δ mutant was defective in macrophage killing in vitro, and was attenuated in virulence in an in vivo Caenorhabditis elegans model representative of intestinal epithelial infection. In contrast, the vps4Δ mutant caused a similar degree of tissue damage in an in vitro uroepithelial model of Candida infection compared with controls. Furthermore, in an in vivo murine model of vaginal candidiasis there was no reduction in fungal colony burden and no differences in vaginal histopathology compared to wild-type and complemented controls. These results suggest that VPS4 contributes to several key aspects of oral epithelial but not uroepithelial infection, and in contrast to systemic infection, plays no major role in the pathogenesis of Candida vaginitis. By using a wide range of virulence models, we demonstrate that C. albicans VPS4 contributes to virulence according to the specific tissue that is infected. Thus, in order to gain a full understanding of C. albicans virulence in relation to a particular gene or pathway of interest, a selected range of infection models may need to be utilized.
Hallie S Rane; Sarah Hardison; Claudia Botelho; Stella M Bernardo; Floyd Wormley; Samuel A Lee. Candida albicansVPS4contributes differentially to epithelial and mucosal pathogenesis. Virulence 2014, 5, 810 -818.
AMA StyleHallie S Rane, Sarah Hardison, Claudia Botelho, Stella M Bernardo, Floyd Wormley, Samuel A Lee. Candida albicansVPS4contributes differentially to epithelial and mucosal pathogenesis. Virulence. 2014; 5 (8):810-818.
Chicago/Turabian StyleHallie S Rane; Sarah Hardison; Claudia Botelho; Stella M Bernardo; Floyd Wormley; Samuel A Lee. 2014. "Candida albicansVPS4contributes differentially to epithelial and mucosal pathogenesis." Virulence 5, no. 8: 810-818.
This study describes a novel liposomal formulation for siRNA delivery, based on the mixture of the neutral lipid mono olein (MO) and cationic lipids of the dioctadecyldimethylammonium (DODA) family. The cationic lipids dioctadecyldimethylammonium bromide (DODAB) and chloride (DODAC) were compared in order to identify which one will most efficiently induce gene silencing. MO has a fluidizing effect on DODAC and DODAB liposomes, although it was more homogeneously distributed in DODAC bilayers. All MO-based liposomal formulations were able to efficiently encapsulate siRNA. Stable lipoplexes of small size (100-160 nm) with a positive surface charge (>+45 mV) were formed. A more uniform MO incorporation in DODAC:MO may explain an increase of the fusogenic potential of these liposomes. The siRNA-lipoplexes were readily internalized by human nonsmall cell lung carcinoma (H1299) cells, in an energy dependent process. DODAB:MO nanocarriers showed a higher internalization efficiency in comparison to DODAC:MO lipoplexes, and were also more efficient in promoting gene silencing. MO had a similar gene silencing ability as the commonly used helper lipid 1,2-dioleyl-3-phosphatidylethanolamine (DOPE), but with much lower cytotoxicity. Taking in consideration all the results presented, DODAB:MO liposomes are the most promising tested formulation for systemic siRNA delivery
Ana Oliveira; Thomas Frans Martens; Koen Raemdonck; Renata Danielle Adati; Eloi Feitosa; Cláudia Botelho; Andreia Gomes; Kevin Braeckmans; Maria Elisabete Cunha Dias Real Oliveira. Dioctadecyldimethylammonium:Monoolein Nanocarriers for Efficient in Vitro Gene Silencing. ACS Applied Materials & Interfaces 2014, 6, 6977 -6989.
AMA StyleAna Oliveira, Thomas Frans Martens, Koen Raemdonck, Renata Danielle Adati, Eloi Feitosa, Cláudia Botelho, Andreia Gomes, Kevin Braeckmans, Maria Elisabete Cunha Dias Real Oliveira. Dioctadecyldimethylammonium:Monoolein Nanocarriers for Efficient in Vitro Gene Silencing. ACS Applied Materials & Interfaces. 2014; 6 (9):6977-6989.
Chicago/Turabian StyleAna Oliveira; Thomas Frans Martens; Koen Raemdonck; Renata Danielle Adati; Eloi Feitosa; Cláudia Botelho; Andreia Gomes; Kevin Braeckmans; Maria Elisabete Cunha Dias Real Oliveira. 2014. "Dioctadecyldimethylammonium:Monoolein Nanocarriers for Efficient in Vitro Gene Silencing." ACS Applied Materials & Interfaces 6, no. 9: 6977-6989.
Biofilms constitute a physical barrier, protecting the encased bacteria from detergents and sanitizers. The objective of this work was to analyze the effectiveness of sodium hypochlorite (NaOCl) against strains of Staphylococcus aureus isolated from raw milk of cows with subclinical mastitis and Staphylococcus aureus isolated from the milking environment (blowers and milk conducting tubes). The results revealed that, in the presence of NaOCl (150ppm), the number of adhered cells of the twelve S. aureus strains was significantly reduced. When the same strains were evaluated in biofilm condition, different results were obtained. It was found that, after a contact period of five minutes with NaOCl (150ppm), four strains (two strains from milk , one from the blowers and one from a conductive rubber) were still able to grow. Although with the increasing contact time between the bacteria and the NaOCl (150ppm), no growth was detected for any of the strains. Concerning the efficiency of NaOCl on total biofilm biomass formation by each S. aureus strain, a decrease was observed when these strains were in contact with 150 ppm NaOCl for a total period of 10 minutes. This study highlights the importance of a correct sanitation protocol of all the milk processing units which can indeed significantly reduce the presence of microorganisms, leading to a decrease of cow´s mastitis and milk contamination.
Poliana De Castro Melo; Cláudia Sousa; Cláudia Botelho; Rosário Oliveira; Antonio Nader-Filho. NaOCl effect on biofilm produced by Staphylococcus aureus isolated from the milking environment and mastitis infected cows. Pesquisa Veterinária Brasileira 2014, 34, 109 -113.
AMA StylePoliana De Castro Melo, Cláudia Sousa, Cláudia Botelho, Rosário Oliveira, Antonio Nader-Filho. NaOCl effect on biofilm produced by Staphylococcus aureus isolated from the milking environment and mastitis infected cows. Pesquisa Veterinária Brasileira. 2014; 34 (2):109-113.
Chicago/Turabian StylePoliana De Castro Melo; Cláudia Sousa; Cláudia Botelho; Rosário Oliveira; Antonio Nader-Filho. 2014. "NaOCl effect on biofilm produced by Staphylococcus aureus isolated from the milking environment and mastitis infected cows." Pesquisa Veterinária Brasileira 34, no. 2: 109-113.