Estabelecimento do perfil metabolómico volátil da urina e saliva, como estratégia não-invasiva, para a deteção de potenciais biomarcadores de diferentes tipos de cancro

Com este trabalho pretendeu-se estabelecer o perfil metabolómico volátil de amostras de fluidos biológicos, nomeadamente saliva e urina, de pacientes com cancro da mama e do pulmão e de indivíduos saudáveis (grupo controlo), utilizando a Microextração em Fase Sólida em modo headspace (HS-SPME) seguida de Cromatografia Gasosa acoplada à Espectrometria de Massa (GC-MS). Efetuou-se a comparação entre os perfis voláteis dos grupos estudados com o objetivo de identificar metabolitos que possam ser considerados como potenciais biomarcadores dos tipos de cancro em estudo. De modo a otimizar a metodologia extrativa, HS-SPME, foram avaliados os diferentes parâmetros experimentais com influência no processo extrativo. Os melhores resultados foram obtidos com a fibra CAR/PDMS, usando um volume de 2 mL de saliva acidificada, 10% NaCl (m/v) e 45 minutos de extração a uma temperatura de 37±1°C. Para a urina foi utilizada a mesma fibra, 4 mL de urina acidificada, 20% NaCl (m/v) e 60 minutos de extração a 50±1°C. Nas amostras de saliva e urina, foram identificados 243 e 500 metabolitos voláteis respetivamente, sendo estes pertencentes a diferentes famílias químicas. Posteriormente, utilizou-se a análise discriminante por mínimos quadrados parciais (PLS-DA) que permitiu observar uma boa separação entre os grupos controlo e oncológicos. Nas amostras salivares o grupo de pacientes com cancro da mama foi maioritariamente caracterizado pelo metabolito ácido benzeno carboxílico e o grupo de pacientes com cancro do pulmão pelo ácido hexanóico. Na urina o grupo de pacientes com cancro da mama foi maioritariamente caracterizado pelo metabolito 1-[2-(Isobutiriloxi)-1-metiletil]-2,2-dimetilpropil 2-metilpropanoato e o grupo de pacientes com cancro do pulmão pelo o-cimeno. Além da metodologia PLS-DA foi realizada a validação cruzada de monte carlo (MCCV) tendo-se obtido uma elevada taxa de classificação, sensibilidade e especificidade o que demonstra a robustez dos dados obtidos

Caracterização dos estilos parentais na toxicodependência

Dissertação de mest., Psicologia, Faculdade de Ciências Humanas e Sociais, Universidade do Algarve, 2007Temas como estilos educativos parentais, família, toxicodependência e sintomas psicopatológicos têm sido alvo de enorme interesse por parte dos clínicos e investigadores; porém, muitos aspectos ainda não são suficientemente conhecidos. Neste estudo, comparam-se dois grupos, um deles constituído por sujeitos toxicodependentes e outro por sujeitos não toxicodependentes; no que se refere à interferência dos estilos parentais, sintomas psicopatológicos e a relação entre estes dois aspectos. Os resultados obtidos distinguem padrões diferentes no que se refere às vivências parentais entre os dois grupos, bem como, diferenças ao nível das diferentes dimensões da psicopatologia

Antimicrobial coating of textiles by laccase in situ polymerization of catechol and p-phenylenediamine

Supplementary data to this article can be found online at https:// doi.org/10.1016/j.reactfunctpolym.2018.11.015.Textile fabrics made up of cotton, wool, and polyethylene terephthalate (PET) were coated with poly(catechol) and poly(p-phenylenediamine) through the in situ enzymatic polymerization of catechol and p-phenylenediamine, assisted by laccase under high-pressure homogenization. All coated fabrics showed antimicrobial activity against both gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) strains and revealed antioxidant character, measured in terms of (ABTSâ¢+)-scavenging activity. The coating of PET and cotton fabrics with both polymers did not affect the viability of foreskin fibroblasts. The methodology proposed for the in situ coating of textile materials assisted by laccase showed to be a promising approach to produce colored antimicrobial textiles with vast potential applications, namely, clothing and medical devices, among others.This study was supported by the Chinese Government Scholarship under China Scholarship Council [grant no. 201606790036], the Chinese Foundation Key Projects of Governmental Cooperation in International Scientific and Technological Innovation [grant no. 2016 YFE0115700], and 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), and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by European Regional Development Fund under the scope of Norte2020 – Programa Operacional Regional do Norte. The work was also supported by the FCT – Fundação para a Ciência e a Tecnologia [grant nº SFRH/BD/121673/2016, SFRH/BPD/98388/2013, and IF/00186/ 2015].info:eu-repo/semantics/publishedVersio

Microextraction by packed sorbent (MEPS) and solid-phase microextraction (SPME) as sample preparation procedures for the metabolomic profiling of urine

For a long time, sample preparation was unrecognized as a critical issue in the analytical methodology, thus limiting the performance that could be achieved. However, the improvement of microextraction techniques, particularly microextraction by packed sorbent (MEPS) and solid-phase microextraction (SPME), completely modified this scenario by introducing unprecedented control over this process. Urine is a biological fluid that is very interesting for metabolomics studies, allowing human health and disease characterization in a minimally invasive form. In this manuscript, we will critically review the most relevant and promising works in this field, highlighting how the metabolomic profiling of urine can be an extremely valuable tool for the early diagnosis of highly prevalent diseases, such as cardiovascular, oncologic and neurodegenerative ones.info:eu-repo/semantics/publishedVersio

NADES-based cork extractives as green ingredients for cosmetics and textiles

The demand for products based on natural ingredients is increasing among cosmetic and textile consumers. Cork extracts contain components of interest with special properties, including antioxidant, anti-inflammatory, and antibacterial activities, that might improve the effectiveness of cosmetic formulations currently on the market and may impart new characteristics to textiles. The main goal of this work was to investigate the effect of the incorporation of three cork extracts into two commercial cosmetic formulations (formulation A and B) and evaluate their role as textile dyeing agents. The extracts (E1, E2, and E3) were obtained from cork powder using natural deep eutectic solvents (NADES) (E1-NADES 1: lactic acid:glycerol, E2-NADES 2: lactic acid:glycine, and E3-NADES 3: lactic acid:sodium citrate) and applied in combination with the solvent. The impact of the extracts on the cosmetic formulations’ properties was evaluated in terms of pH, viscosity, antioxidant activity, transdermal permeation capacity, cytotoxicity, and organoleptic characteristics (odor, color, and appearance). The results demonstrated that the cork extracts improved the antioxidant performance of the formulations (90% reduction in DPPH (1,1-difenil-2-picril-hidrazil)). Moreover, low concentrations (5 mg/mL and 10 mg/mL) of extract did not present a cytotoxic effect on keratinocytes. Cotton fabrics were efficiently dyed with the NADES-based cork extracts which conferred to these substrates antioxidant (78% in DPPH reduction) and antibacterial abilities (inhibition halos: 12–15 mm). The application of cork extracts as ingredients in cosmetics or as dyeing/coloration agents for textile coloration is revealed to be a promising and green route to replace harmful ingredients normally used in industry.This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit. D.S.F. also thanks the Portuguese Foundation for Science and Technology (FCT) for funding (SFRH/BD/147190/2019). A.R. thanks to Portuguese Foundation for Science and Technology (FCT) for is contract under the CEEC-Individual—4th Edition with the reference 2021.02803.CEECIND.info:eu-repo/semantics/publishedVersio

Breath analysis as a potential and non-invasive frontier in disease diagnosis: an overview

Currently, a small number of diseases, particularly cardiovascular (CVDs), oncologic (ODs), neurodegenerative (NDDs), chronic respiratory diseases, as well as diabetes, form a severe burden to most of the countries worldwide. Hence, there is an urgent need for development of efficient diagnostic tools, particularly those enabling reliable detection of diseases, at their early stages, preferably using non-invasive approaches. Breath analysis is a non-invasive approach relying only on the characterisation of volatile composition of the exhaled breath (EB) that in turn reflects the volatile composition of the bloodstream and airways and therefore the status and condition of the whole organism metabolism. Advanced sampling procedures (solid-phase and needle traps microextraction) coupled with modern analytical technologies (proton transfer reaction mass spectrometry, selected ion flow tube mass spectrometry, ion mobility spectrometry, e-noses, etc.) allow the characterisation of EB composition to an unprecedented level. However, a key challenge in EB analysis is the proper statistical analysis and interpretation of the large and heterogeneous datasets obtained from EB research. There is no standard statistical framework/protocol yet available in literature that can be used for EB data analysis towards discovery of biomarkers for use in a typical clinical setup. Nevertheless, EB analysis has immense potential towards development of biomarkers for the early disease diagnosis of diseases.info:eu-repo/semantics/publishedVersio

Screening of salivary volatiles for putative breast cancer discrimination: an exploratory study involving geographically distant populations

Saliva is possibly the easiest biofluid to analyse and, despite its simple composition, contains relevant metabolic information. In this work, we explored the potential of the volatile composition of saliva samples as biosignatures for breast cancer (BC) non-invasive diagnosis. To achieve this, 106 saliva samples of BC patients and controls in two distinct geographic regions in Portugal and India were extracted and analysed using optimised headspace solid-phase microextraction gas chromatography mass spectrometry (HS-SPME/GC-MS, 2 mL acidified saliva containing 10% NaCl, stirred (800 rpm) for 45 min at 38 °C and using the CAR/PDMS SPME fibre) followed by multivariate statistical analysis (MVSA). Over 120 volatiles from distinct chemical classes, with significant variations among the groups, were identified. MVSA retrieved a limited number of volatiles, viz. 3-methyl-pentanoic acid, 4-methyl-pentanoic acid, phenol and p-tert-butyl-phenol (Portuguese samples) and acetic, propanoic, benzoic acids, 1,2-decanediol, 2-decanone, and decanal (Indian samples), statistically relevant for the discrimination of BC patients in the populations analysed. This work defines an experimental layout, HS-SPME/GC-MS followed by MVSA, suitable to characterise volatile fingerprints for saliva as putative biosignatures for BC non-invasive diagnosis. Here, it was applied to BC samples from geographically distant populations and good disease separation was obtained. Further studies using larger cohorts are therefore very pertinent to challenge and strengthen this proof-of-concept study.info:eu-repo/semantics/publishedVersio

Cryptic susceptibility to penicillin/β-lactamase inhibitor combinations in emerging multidrug-resistant, hospital-adapted Staphylococcus epidermidis lineages.

Acknowledgements: This work was supported by the Wellcome Trust under Grant 220540/Z/20/A (E.M.H.). We thank Frederikke Rosenborg Petersen and Emilie Due Jensen for technical assistance during the animal experiments.Global spread of multidrug-resistant, hospital-adapted Staphylococcus epidermidis lineages underscores the need for new therapeutic strategies. Here we show that many S. epidermidis isolates belonging to these lineages display cryptic susceptibility to penicillin/β-lactamase inhibitor combinations under in vitro conditions, despite carrying the methicillin resistance gene mecA. Using a mouse thigh model of S. epidermidis infection, we demonstrate that single-dose treatment with amoxicillin/clavulanic acid significantly reduces methicillin-resistant S. epidermidis loads without leading to detectable resistance development. On the other hand, we also show that methicillin-resistant S. epidermidis is capable of developing increased resistance to amoxicillin/clavulanic acid during long-term in vitro exposure to these drugs. These findings suggest that penicillin/β-lactamase inhibitor combinations could be a promising therapeutic candidate for treatment of a high proportion of methicillin-resistant S. epidermidis infections, although the in vivo risk of resistance development needs to be further addressed before they can be incorporated into clinical trials