Quercus suber L. bark (cork) is a sustainable material due to its ability to regenerate. The aim ... more Quercus suber L. bark (cork) is a sustainable material due to its ability to regenerate. The aim of this work was to explore cork powders, by-products of the cork industry, as sustainable sources of value-added compounds. Two types of cork powder were studied: coarse (P0) and fine (P1). A broad physicochemical characterization was carried out, regarding particle size, color, moisture content, hygroscopicity, pH, heavy metal content, NIR spectra, and volatile compounds. DPPH scavenging activity and total phenolic content were also evaluated for an ethanolic P1 extract. For both powders, Hg, As, Cd and Pb contents were below the acceptable limits and volatile compounds commonly used as fragrances were found in their composition. P1 had a smaller and more homogenous particle size, lighter brownish color, lower pH value, and lower moisture content than P0, and therefore appears to be more suitable for industrial application. P1 ethanolic extract also showed a high scavenging activity an...
The manganese(III) porphyrin MnTnHex-2-PyP5+ (MnTnHex) is a potent superoxide dismutase mimic and... more The manganese(III) porphyrin MnTnHex-2-PyP5+ (MnTnHex) is a potent superoxide dismutase mimic and modulator of redox-based transcriptional activity that has been studied in the context of different human disease models, including cancer. Nevertheless, for lung cancer, hardly any information is available. Thus, the present work aims to fill this gap and reports the effects of MnTnHex in non-small cell lung cancer (NSCLC) cells, more specifically, A549 and H1975 cells, in vitro. Both cell lines were initially characterized in terms of innate levels of catalase, glutathione peroxidase 1, and peroxiredoxins 1 and 2. To assess the effect of MnTnHex in NSCLC, alone or in combination with cisplatin, endpoints related to the cell viability, cell cycle distribution, cell motility, and characterization of the volatile carbonyl compounds (VCCs) generated in the extracellular medium (i.e., exometabolome) were addressed. The results show that MnTnHex as a single drug markedly reduced the viabili...
Sunitinib and pazopanib are tyrosine kinase inhibitors (TKIs) used as first-line therapy for meta... more Sunitinib and pazopanib are tyrosine kinase inhibitors (TKIs) used as first-line therapy for metastatic renal cell carcinoma (RCC). Although these TKIs are associated with similar survival outcomes, some differences have been reported in their safety profiles. In this work, traditional toxicological endpoints (cell viability and growth, oxidative stress, and nuclear morphology) and 1H NMR spectroscopy-based metabolomics analysis were used to provide new insights into the cytotoxicity and metabolic mechanisms underlying sunitinib and pazopanib treatments. Tumoral (Caki-1) and non-tumoral (HK-2) human renal cells were exposed to clinically relevant concentrations of sunitinib (2 µM) or pazopanib (50 µM). Sunitinib showed selectivity for cancer cells, inhibiting proliferation, and inducing apoptotic death of Caki-1 cells, whereas pazopanib had a similar cytotoxic effect in both tumoral and non-tumoral cells. 1H-NMR metabolomics unveiled a higher impact of sunitinib on the levels of int...
2º Encontro Nacional de Jovens Investigadores em Oncologia (ENJIO 2020), Sep 24, 2020
O carcinoma de células renais do subtipo células claras (ccCCR) representa o tipo mais comum (~70... more O carcinoma de células renais do subtipo células claras (ccCCR) representa o tipo mais comum (~70%) de cancro renal. Investigações recentes sugerem que o desenvolvimento deste tipo de cancro está relacionado com alterações metabólicas induzidas por mutações que ocorrem em genes que controlam o metabolismo celular (p.ex., von Hippel–Lindau e fumarato hidratase). Deste modo, o estudo de potenciais biomarcadores de ccCCR, através de uma abordagem metabolómica, torna-se bastante pertinente e atual. Em particular, a fração volátil do metaboloma urinário tem revelado resultados muito promissores na identificação de painéis de biomarcadores com elevada sensibilidade para a deteção de cancros urológicos. Devido aos avanços consideráveis na área dos sensores bioelectrónicos, espera-se que num futuro próximo este tipo de cancro possa ser diagnosticado de uma forma simples, rápida e não invasiva em contexto clínico. Objetivos: Este trabalho teve como objetivo principal a identificação de um painel de biomarcadores para deteção não invasiva de ccCCR através da análise do perfil volátil da urina de doentes com ccCCR (n=75) e de indivíduos controlo (sem cancro, n=75). Material e Métodos: Os compostos orgânicos voláteis (COVs), e mais especificamente os compostos carbonílicos voláteis (CCVs), presentes na urina foram analisados por microextração em fase sólida por headspace e cromatografia gasosa acoplada à espectrometria de massa (HS-SPME-GC-MS). Os dados obtidos foram pré-processados e analisados com recurso a ferramentas bioinformáticas. Resultados: Os modelos de classificação obtidos para as análises de COVs e CCVs mostraram uma boa separação entre as urinas de doentes com ccCCR e controlos, com áreas sob a curva (AUC) de 0,846 e 0,818, respetivamente. No total, 22 compostos revelaram alterações estatisticamente significativas entre os dois grupos, incluindo vários aldeídos, cetonas, hidrocarbonetos aromáticos e terpenos. Foi ainda encontrado um conjunto de seis potenciais biomarcadores de ccCCR que revelou uma AUC de 0.869, sensibilidade de 80%, especificidade de 82% e exatidão de 81%. Deste painel fazem parte o octanal, 3-metilbutanal, benzaldeído, 2-furaldeído, 4-heptanona e p-cresol. As desregulações observadas nos níveis destes compostos sugerem alterações no metabolismo energético e uma maior expressão das enzimas ligadas a processos de carcinogénese. Conclusões: Estes resultados confirmam o potencial da assinatura volátil da urina para a deteção não invasiva de ccCCR e revelam um conjunto de biomarcadores moleculares que podem ser utilizados no desenvolvimento de novos materiais que permitam o seu reconhecimento por sensores bioelectrónicos, comummente chamados de “E-noses”.info:eu-repo/semantics/publishedVersio
Renal Cell Carcinoma (RCC) constitutes approximately 90-95% of all kidney neoplasms and is the se... more Renal Cell Carcinoma (RCC) constitutes approximately 90-95% of all kidney neoplasms and is the second most lethal urological cancer. Current diagnostic techniques rely on imaging techniques and an invasive procedure (biopsy) is always required for histopathologic confirmation of malignancy. For these reasons, the identification of accurate biomarkers to develop faster, less invasive and more sophisticated diagnostic techniques is of utmost importance. Metabolomics has been widely applied in cancer biomarker discovery arising from the fact that cancer cells are metabolically reprogrammed to control the energy required by the rapid growth and development of the tumor, producing a specific “metabolic signature”. Aim: To evaluate the potential of volatile organic compounds (VOCs) and volatile carbonyl compounds (VCCs) to discriminate the exometabolome of RCC from non-tumoral cell lines, and two different histological subtypes (clear cell and papillary RCC) in both metastatic and non-metastatic stages. Methods: Headspace-solid phase microextraction/gas chromatography-mass spectrometry (HS-SPME/GC-MS)-based metabolomics was applied for the volatile profiling of culture medium of five different tumoral cell lines, namely three clear cell (769-P, 786-O and Caki-1) and two papillary RCC (Caki-2 and ACHN), and one non-tumoral cell line (HK-2). Results: Multivariate and univariate analysis unveiled a panel of metabolites responsible for the discrimination between each RCC cell line vs. non-tumoral cells, metastatic vs. non-metastatic and clear cell vs. papillary RCC cell lines, mostly belonging to alcohols, aldehydes, alkanes and ketones classes. Some metabolites were found similarly altered for all RCC cell lines compared to the non-tumoral, while others unveiled specificity for each RCC cell line. Discussion/Conclusion: The volatile exometabolome signature of RCC cells can provide candidate biomarkers for the development of a volatile sensor-based approach for non-invasive diagnosis of RCC in urine.info:eu-repo/semantics/publishedVersio
2º Encontro Nacional de Jovens Investigadores em Oncologia (ENJIO 2020), Sep 24, 2020
O Carcinoma de Células Renais (CCR) representa o terceiro cancro urológico mais frequente e letal... more O Carcinoma de Células Renais (CCR) representa o terceiro cancro urológico mais frequente e letal em Portugal. Este tipo de cancro inclui vários subtipos histológicos com diferentes potenciais metastáticos, sendo o carcinoma de células claras (ccCCR) e o papilar (pCCR) os mais comuns. É atualmente reconhecida a importância da descoberta de biomarcadores moleculares específicos para o diagnóstico e estadiamento do CCR de forma a superar as limitações dos métodos existentes (ecografia, tomografia computorizada e nefrectomia). O uso da metabolómica constitui uma das abordagens mais promissoras para a identificação de biomarcadores, uma vez que o metabolismo das células tumorais é muito diferente do das células normais. Desta forma, estudos recentes têm mostrado o potencial dos compostos orgânicos voláteis (COVs) e compostos carbonílicos voláteis (CCVs), não só na identificação de novos biomarcadores, mas também na compreensão de vias metabólicas envolvidas na carcinogénese. Objetivos: Este trabalho incluiu o estudo do exometaboloma volátil de linhas celulares dos dois subtipos histológicos mais comuns de CCR (ccCCR e pCCR) com diferentes potenciais metastáticos. Assim, o objetivo principal deste trabalho é identificar os COVs e CCVs, presentes no meio extracelular (exometaboloma), com potencial para serem utilizados como biomarcadores no processo de estadiamento de CCR e consequentemente ajudar na escolha da terapêutica mais dirigida. Material e Métodos: De modo a avaliar as diferenças na composição volátil do exometaboloma de três linhas celulares de ccCCR (não metastáticas: 769-P, 786-O; metastáticas: Caki-1) e duas de pCCR (não metastática: Caki-2; metastática: ACHN), os COVs e CCVs foram extraídos no headspace do meio de cultura por microextração em fase sólida (HS-SPME) e analisados por cromatografia gasosa acoplada à espectrometria de massa (GC-MS). Os dados foram posteriormente tratados através de métodos de análise estatística multivariada e univariada. Resultados: Os resultados obtidos demonstraram que o exometaboloma volátil é mais robusto a discriminar células com diferentes potenciais metastáticos do que a discriminar entre os dois subtipos histológicos. No entanto, foram encontradas diferenças significativas nos níveis de quinze compostos entre ccCCR e pCCR, tais como o 2,4-dimetil-1-hepteno, 3-careno e 4-metilbenzaldeído. Na comparação entre as linhas celulares metastáticas e não metastáticas de ccCCR, foram detetadas alterações nos níveis de diversos alcanos, alcenos e derivados de benzeno, enquanto que nas linhas celulares pCCR, as principais alterações encontradas foram em compostos pertencentes à classe dos aldeídos e cetonas, como o acetaldeído e a 2-pentadecanona. Conclusões: Os resultados obtidos revelaram o potencial do exometaboloma volátil na identificação de biomarcadores promissores para o diagnóstico e estadiamento de CCR.info:eu-repo/semantics/publishedVersio
Wine flavor undergoes major changes during bottle aging and can be influenced by the type of clos... more Wine flavor undergoes major changes during bottle aging and can be influenced by the type of closure. The interaction between wine, the type of closure and the external environment has the potential to significantly influence the overall quality of bottled wines, especially when the storage period is relatively long (more than five years). Therefore, the choice of closure (cork, synthetic or screw cap) deserves special attention in order to establish the ideal sealing conditions for optimizing wine flavor attributes. The contribution of different closures to the quality of bottled wine is through mass transfer phenomena, including permeation, sorption (scalping) or desorption of chemicals between closure materials and wines. Thus, this article aims to review the impact of different closures on the flavor composition of wines during post-bottling conditions. The implications of closures on wine sensory properties are also discussed.
Large amounts of Prunus avium L. by-products result from sweet cherry production and processing. ... more Large amounts of Prunus avium L. by-products result from sweet cherry production and processing. This work aimed to evaluate the mineral content and volatile profiling of the cherry stems, leaves, and flowers of the Saco cultivar collected from the Fundão region (Portugal). A total of 18 minerals were determined by ICP-MS, namely 8 essential and 10 non-essential elements. Phosphorus (P) was the most abundant mineral, while lithium (Li) was detected in trace amounts. Three different preparations were used in this work to determine volatiles: hydroethanolic extracts, crude extracts, and aqueous infusions. A total of 117 volatile compounds were identified using HS-SPME/GC-MS, distributed among different chemical classes: 31 aldehydes, 14 alcohols, 16 ketones, 30 esters, 4 acids, 4 monoterpenes, 3 norisoprenoids, 4 hydrocarbons, 7 heterocyclics, 1 lactone, 1 phenol, and 2 phenylpropenes. Benzaldehyde, 4-methyl-benzaldehyde, hexanal, lilac aldehyde, and 6-methyl-5-hepten-2-one were the m...
Prostate cancer (PCa), bladder cancer (BCa), and renal cell carcinoma (RCC) are the most common u... more Prostate cancer (PCa), bladder cancer (BCa), and renal cell carcinoma (RCC) are the most common urological cancers, and their incidence has been rising over time. Surgery is the standard treatment for these cancers, but this procedure is only effective when the disease is localized. For metastatic disease, PCa is typically treated with androgen deprivation therapy, while BCa is treated with chemotherapy, and RCC is managed primarily with targeted therapies. However, response rates to these therapeutic options remain unsatisfactory due to the development of resistance and treatment-related toxicity. Thus, the discovery of biomarkers with prognostic and predictive value is needed to stratify patients into different risk groups, minimizing overtreatment and the risk of drug resistance development. Pharmacometabolomics, a branch of metabolomics, is an attractive tool to predict drug response in an individual based on its own metabolic signature, which can be collected before, during, an...
This study aimed at investigating the impact of different technical cork stoppers on the quality ... more This study aimed at investigating the impact of different technical cork stoppers on the quality preservation and shelf life of sparkling wines. The volatile compositions of two Italian sparkling wines sealed with a sparkling cork with two natural cork discs (2D) and a microagglomerated (MA) cork were determined during bottle aging (12 to 42 months) after disgorging, by headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC/MS). The volatile profile of the sparkling wine #1 sealed with 2D stoppers showed the presence of camphor from 12 to 42 months, along with significant alterations in the levels of several alcohols, ketones, and ethyl esters at 24 and 42 months. The impact of closure type was less pronounced for sparkling wine #2 which also showed the presence of camphor from 12 to 42 months in 2D samples, and significantly higher levels of esters at 24 and 42 months for 2D compared with MA. These results unveiled that the type of closu...
The main purpose of this work was to describe and validate a new methodology for the determinatio... more The main purpose of this work was to describe and validate a new methodology for the determination of a brandy reactivity index (BRI) towards flavan-3-ol compounds in wines (catechins and condensed tannins) and to correlate this index with the amount and type of aldehydes present in those brandies. This new method consists of two main reactions: a) the ability of aldehydes to react with C8 and/or C6 positions in the phloroglucinol ring of a catechin present in excess (A ring); b) further reaction of the remaining free catechin with p-dimethylaminocinamaldehyde (DMACA) yielding to the formation of a blue compound that can be quantified by Visible spectroscopy at 640 nm. The impact of different experimental conditions such as reagent concentrations and reaction time on BRI were also evaluated. The method was validated through the determination of repeatability (intra-day variability) and reproducibility. The repeatability was considered acceptable with a CV of 11.87 %. The analysis of...
Adrenaline, which participates in the neuroendocrine response that occurs during stress and perim... more Adrenaline, which participates in the neuroendocrine response that occurs during stress and perimenopause, may be tumorigenic. This exploratory study aimed at investigating whether non-tumorigenic and tumorigenic human breast epithelial cell lines are able to synthesize adrenaline. The study was carried out in non-tumorigenic (MCF-10A) and tumorigenic (MCF-7) human breast cell lines. Expression of enzymes involved in adrenaline synthesis was characterized by RT-qPCR, immunocytochemistry and western blot. Catecholamines and analogue compounds were quantified by HPLC-ECD. Functional assessment of the impact of drugs on cells’ tumorigenic potential was assessed by determination of cell viability and clonogenic ability. Both MCF-10A and MCF-7 cells produce catecholamines, but the capacity to produce adrenaline is lower in MCF-10A cells. β-adrenoceptor activation increases the capacity of MCF-10A cells to produce adrenaline and favor both cell viability and colony formation. It is conclu...
Prostate cancer (PCa) is the second most diagnosed cancer in men worldwide. For its screening, se... more Prostate cancer (PCa) is the second most diagnosed cancer in men worldwide. For its screening, serum prostate specific antigen (PSA) test has been largely performed over the past decade, despite its lack of accuracy and inability to distinguish indolent from aggressive disease. Metabolomics has been widely applied in cancer biomarker discovery due to the well-known metabolic reprogramming characteristic of cancer cells. Most of the metabolomic studies have reported alterations in urine of PCa patients due its noninvasive collection, but the analysis of prostate tissue metabolome is an ideal approach to disclose specific modifications in PCa development. This review aims to summarize and discuss the most recent findings from tissue and urine metabolomic studies applied to PCa biomarker discovery. Eighteen metabolites were found consistently altered in PCa tissue among different studies, including alanine, arginine, uracil, glutamate, fumarate, and citrate. Urine metabolomic studies a...
An untargeted approach, focused in the profile of volatile organic compounds (VOCs), was applied ... more An untargeted approach, focused in the profile of volatile organic compounds (VOCs), was applied to differentiate natural cork stoppers with different levels of porosity, coded as Group 1 (low porosity), Group 2 (intermediate porosity) and Group 3 (high porosity). Statistically significant alterations were found in the levels of several VOCs between cork stoppers of low and intermediate porosity when compared with those of high porosity (Group 1 vs. 3 and Group 2 vs. 3). In addition, the levels of 2-pentylfuran, cyclene, camphene, camphor, limonene and eucalyptol enabled the discrimination of cork stoppers with low porosity (Group 1) into two subgroups, while furfural and 5-methyl-2-furfural enabled the discrimination of subgroups within the intermediate and high porosity stoppers (Group 2 and 3). A headspace solid-phase microextraction coupled to gas chromatography tandem mass spectrometry (HS-SPME-GC-MS/MS) method was developed to quantify the subgroup discriminant compounds, which may provide a proof-of-concept for the development of an efficient method to be applied in cork industry.
Clear cell renal cell carcinoma (ccRCC) is the most common type of kidney cancer usually associat... more Clear cell renal cell carcinoma (ccRCC) is the most common type of kidney cancer usually associated with asymptomatic development and risk of systemic progression. Hence, reliable molecular biomarkers of ccRCC are needed to provide early and minimally invasive detection. In this study, urinary volatilome profiling of patients diagnosed with ccRCC (n = 75), and cancer-free controls (n = 75), was performed to investigate the presence of a volatile signature characteristic of ccRCC. Volatile organic compounds (VOCs) in general, and more specifically volatile carbonyl compounds (VCCs), present in urine were extracted by headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS). Supervised multivariate models showed a good discriminatory power of ccRCC patients from controls in urine. Overall, 22 volatile metabolites were found significantly altered between the two groups, including aldehydes, ketones, aromatic hydrocarbons, and terpenoids. A candidate six-biomarker panel, comprising octanal, 3-methylbutanal, benzaldehyde, 2-furaldehyde, 4-heptanone, and p-cresol, depicted the best performance for ccRCC detection with 83% sensitivity, 79% specificity, and 81% accuracy. Moreover, the ccRCC urinary volatilome signature suggested dysregulation of energy metabolism and overexpression of enzymes associated with carcinogenesis. These findings provide the molecular basis for the fine-tuning of gas-sensing materials for application in the development of a bioelectronic sensor.
Quercus suber L. bark (cork) is a sustainable material due to its ability to regenerate. The aim ... more Quercus suber L. bark (cork) is a sustainable material due to its ability to regenerate. The aim of this work was to explore cork powders, by-products of the cork industry, as sustainable sources of value-added compounds. Two types of cork powder were studied: coarse (P0) and fine (P1). A broad physicochemical characterization was carried out, regarding particle size, color, moisture content, hygroscopicity, pH, heavy metal content, NIR spectra, and volatile compounds. DPPH scavenging activity and total phenolic content were also evaluated for an ethanolic P1 extract. For both powders, Hg, As, Cd and Pb contents were below the acceptable limits and volatile compounds commonly used as fragrances were found in their composition. P1 had a smaller and more homogenous particle size, lighter brownish color, lower pH value, and lower moisture content than P0, and therefore appears to be more suitable for industrial application. P1 ethanolic extract also showed a high scavenging activity an...
The manganese(III) porphyrin MnTnHex-2-PyP5+ (MnTnHex) is a potent superoxide dismutase mimic and... more The manganese(III) porphyrin MnTnHex-2-PyP5+ (MnTnHex) is a potent superoxide dismutase mimic and modulator of redox-based transcriptional activity that has been studied in the context of different human disease models, including cancer. Nevertheless, for lung cancer, hardly any information is available. Thus, the present work aims to fill this gap and reports the effects of MnTnHex in non-small cell lung cancer (NSCLC) cells, more specifically, A549 and H1975 cells, in vitro. Both cell lines were initially characterized in terms of innate levels of catalase, glutathione peroxidase 1, and peroxiredoxins 1 and 2. To assess the effect of MnTnHex in NSCLC, alone or in combination with cisplatin, endpoints related to the cell viability, cell cycle distribution, cell motility, and characterization of the volatile carbonyl compounds (VCCs) generated in the extracellular medium (i.e., exometabolome) were addressed. The results show that MnTnHex as a single drug markedly reduced the viabili...
Sunitinib and pazopanib are tyrosine kinase inhibitors (TKIs) used as first-line therapy for meta... more Sunitinib and pazopanib are tyrosine kinase inhibitors (TKIs) used as first-line therapy for metastatic renal cell carcinoma (RCC). Although these TKIs are associated with similar survival outcomes, some differences have been reported in their safety profiles. In this work, traditional toxicological endpoints (cell viability and growth, oxidative stress, and nuclear morphology) and 1H NMR spectroscopy-based metabolomics analysis were used to provide new insights into the cytotoxicity and metabolic mechanisms underlying sunitinib and pazopanib treatments. Tumoral (Caki-1) and non-tumoral (HK-2) human renal cells were exposed to clinically relevant concentrations of sunitinib (2 µM) or pazopanib (50 µM). Sunitinib showed selectivity for cancer cells, inhibiting proliferation, and inducing apoptotic death of Caki-1 cells, whereas pazopanib had a similar cytotoxic effect in both tumoral and non-tumoral cells. 1H-NMR metabolomics unveiled a higher impact of sunitinib on the levels of int...
2º Encontro Nacional de Jovens Investigadores em Oncologia (ENJIO 2020), Sep 24, 2020
O carcinoma de células renais do subtipo células claras (ccCCR) representa o tipo mais comum (~70... more O carcinoma de células renais do subtipo células claras (ccCCR) representa o tipo mais comum (~70%) de cancro renal. Investigações recentes sugerem que o desenvolvimento deste tipo de cancro está relacionado com alterações metabólicas induzidas por mutações que ocorrem em genes que controlam o metabolismo celular (p.ex., von Hippel–Lindau e fumarato hidratase). Deste modo, o estudo de potenciais biomarcadores de ccCCR, através de uma abordagem metabolómica, torna-se bastante pertinente e atual. Em particular, a fração volátil do metaboloma urinário tem revelado resultados muito promissores na identificação de painéis de biomarcadores com elevada sensibilidade para a deteção de cancros urológicos. Devido aos avanços consideráveis na área dos sensores bioelectrónicos, espera-se que num futuro próximo este tipo de cancro possa ser diagnosticado de uma forma simples, rápida e não invasiva em contexto clínico. Objetivos: Este trabalho teve como objetivo principal a identificação de um painel de biomarcadores para deteção não invasiva de ccCCR através da análise do perfil volátil da urina de doentes com ccCCR (n=75) e de indivíduos controlo (sem cancro, n=75). Material e Métodos: Os compostos orgânicos voláteis (COVs), e mais especificamente os compostos carbonílicos voláteis (CCVs), presentes na urina foram analisados por microextração em fase sólida por headspace e cromatografia gasosa acoplada à espectrometria de massa (HS-SPME-GC-MS). Os dados obtidos foram pré-processados e analisados com recurso a ferramentas bioinformáticas. Resultados: Os modelos de classificação obtidos para as análises de COVs e CCVs mostraram uma boa separação entre as urinas de doentes com ccCCR e controlos, com áreas sob a curva (AUC) de 0,846 e 0,818, respetivamente. No total, 22 compostos revelaram alterações estatisticamente significativas entre os dois grupos, incluindo vários aldeídos, cetonas, hidrocarbonetos aromáticos e terpenos. Foi ainda encontrado um conjunto de seis potenciais biomarcadores de ccCCR que revelou uma AUC de 0.869, sensibilidade de 80%, especificidade de 82% e exatidão de 81%. Deste painel fazem parte o octanal, 3-metilbutanal, benzaldeído, 2-furaldeído, 4-heptanona e p-cresol. As desregulações observadas nos níveis destes compostos sugerem alterações no metabolismo energético e uma maior expressão das enzimas ligadas a processos de carcinogénese. Conclusões: Estes resultados confirmam o potencial da assinatura volátil da urina para a deteção não invasiva de ccCCR e revelam um conjunto de biomarcadores moleculares que podem ser utilizados no desenvolvimento de novos materiais que permitam o seu reconhecimento por sensores bioelectrónicos, comummente chamados de “E-noses”.info:eu-repo/semantics/publishedVersio
Renal Cell Carcinoma (RCC) constitutes approximately 90-95% of all kidney neoplasms and is the se... more Renal Cell Carcinoma (RCC) constitutes approximately 90-95% of all kidney neoplasms and is the second most lethal urological cancer. Current diagnostic techniques rely on imaging techniques and an invasive procedure (biopsy) is always required for histopathologic confirmation of malignancy. For these reasons, the identification of accurate biomarkers to develop faster, less invasive and more sophisticated diagnostic techniques is of utmost importance. Metabolomics has been widely applied in cancer biomarker discovery arising from the fact that cancer cells are metabolically reprogrammed to control the energy required by the rapid growth and development of the tumor, producing a specific “metabolic signature”. Aim: To evaluate the potential of volatile organic compounds (VOCs) and volatile carbonyl compounds (VCCs) to discriminate the exometabolome of RCC from non-tumoral cell lines, and two different histological subtypes (clear cell and papillary RCC) in both metastatic and non-metastatic stages. Methods: Headspace-solid phase microextraction/gas chromatography-mass spectrometry (HS-SPME/GC-MS)-based metabolomics was applied for the volatile profiling of culture medium of five different tumoral cell lines, namely three clear cell (769-P, 786-O and Caki-1) and two papillary RCC (Caki-2 and ACHN), and one non-tumoral cell line (HK-2). Results: Multivariate and univariate analysis unveiled a panel of metabolites responsible for the discrimination between each RCC cell line vs. non-tumoral cells, metastatic vs. non-metastatic and clear cell vs. papillary RCC cell lines, mostly belonging to alcohols, aldehydes, alkanes and ketones classes. Some metabolites were found similarly altered for all RCC cell lines compared to the non-tumoral, while others unveiled specificity for each RCC cell line. Discussion/Conclusion: The volatile exometabolome signature of RCC cells can provide candidate biomarkers for the development of a volatile sensor-based approach for non-invasive diagnosis of RCC in urine.info:eu-repo/semantics/publishedVersio
2º Encontro Nacional de Jovens Investigadores em Oncologia (ENJIO 2020), Sep 24, 2020
O Carcinoma de Células Renais (CCR) representa o terceiro cancro urológico mais frequente e letal... more O Carcinoma de Células Renais (CCR) representa o terceiro cancro urológico mais frequente e letal em Portugal. Este tipo de cancro inclui vários subtipos histológicos com diferentes potenciais metastáticos, sendo o carcinoma de células claras (ccCCR) e o papilar (pCCR) os mais comuns. É atualmente reconhecida a importância da descoberta de biomarcadores moleculares específicos para o diagnóstico e estadiamento do CCR de forma a superar as limitações dos métodos existentes (ecografia, tomografia computorizada e nefrectomia). O uso da metabolómica constitui uma das abordagens mais promissoras para a identificação de biomarcadores, uma vez que o metabolismo das células tumorais é muito diferente do das células normais. Desta forma, estudos recentes têm mostrado o potencial dos compostos orgânicos voláteis (COVs) e compostos carbonílicos voláteis (CCVs), não só na identificação de novos biomarcadores, mas também na compreensão de vias metabólicas envolvidas na carcinogénese. Objetivos: Este trabalho incluiu o estudo do exometaboloma volátil de linhas celulares dos dois subtipos histológicos mais comuns de CCR (ccCCR e pCCR) com diferentes potenciais metastáticos. Assim, o objetivo principal deste trabalho é identificar os COVs e CCVs, presentes no meio extracelular (exometaboloma), com potencial para serem utilizados como biomarcadores no processo de estadiamento de CCR e consequentemente ajudar na escolha da terapêutica mais dirigida. Material e Métodos: De modo a avaliar as diferenças na composição volátil do exometaboloma de três linhas celulares de ccCCR (não metastáticas: 769-P, 786-O; metastáticas: Caki-1) e duas de pCCR (não metastática: Caki-2; metastática: ACHN), os COVs e CCVs foram extraídos no headspace do meio de cultura por microextração em fase sólida (HS-SPME) e analisados por cromatografia gasosa acoplada à espectrometria de massa (GC-MS). Os dados foram posteriormente tratados através de métodos de análise estatística multivariada e univariada. Resultados: Os resultados obtidos demonstraram que o exometaboloma volátil é mais robusto a discriminar células com diferentes potenciais metastáticos do que a discriminar entre os dois subtipos histológicos. No entanto, foram encontradas diferenças significativas nos níveis de quinze compostos entre ccCCR e pCCR, tais como o 2,4-dimetil-1-hepteno, 3-careno e 4-metilbenzaldeído. Na comparação entre as linhas celulares metastáticas e não metastáticas de ccCCR, foram detetadas alterações nos níveis de diversos alcanos, alcenos e derivados de benzeno, enquanto que nas linhas celulares pCCR, as principais alterações encontradas foram em compostos pertencentes à classe dos aldeídos e cetonas, como o acetaldeído e a 2-pentadecanona. Conclusões: Os resultados obtidos revelaram o potencial do exometaboloma volátil na identificação de biomarcadores promissores para o diagnóstico e estadiamento de CCR.info:eu-repo/semantics/publishedVersio
Wine flavor undergoes major changes during bottle aging and can be influenced by the type of clos... more Wine flavor undergoes major changes during bottle aging and can be influenced by the type of closure. The interaction between wine, the type of closure and the external environment has the potential to significantly influence the overall quality of bottled wines, especially when the storage period is relatively long (more than five years). Therefore, the choice of closure (cork, synthetic or screw cap) deserves special attention in order to establish the ideal sealing conditions for optimizing wine flavor attributes. The contribution of different closures to the quality of bottled wine is through mass transfer phenomena, including permeation, sorption (scalping) or desorption of chemicals between closure materials and wines. Thus, this article aims to review the impact of different closures on the flavor composition of wines during post-bottling conditions. The implications of closures on wine sensory properties are also discussed.
Large amounts of Prunus avium L. by-products result from sweet cherry production and processing. ... more Large amounts of Prunus avium L. by-products result from sweet cherry production and processing. This work aimed to evaluate the mineral content and volatile profiling of the cherry stems, leaves, and flowers of the Saco cultivar collected from the Fundão region (Portugal). A total of 18 minerals were determined by ICP-MS, namely 8 essential and 10 non-essential elements. Phosphorus (P) was the most abundant mineral, while lithium (Li) was detected in trace amounts. Three different preparations were used in this work to determine volatiles: hydroethanolic extracts, crude extracts, and aqueous infusions. A total of 117 volatile compounds were identified using HS-SPME/GC-MS, distributed among different chemical classes: 31 aldehydes, 14 alcohols, 16 ketones, 30 esters, 4 acids, 4 monoterpenes, 3 norisoprenoids, 4 hydrocarbons, 7 heterocyclics, 1 lactone, 1 phenol, and 2 phenylpropenes. Benzaldehyde, 4-methyl-benzaldehyde, hexanal, lilac aldehyde, and 6-methyl-5-hepten-2-one were the m...
Prostate cancer (PCa), bladder cancer (BCa), and renal cell carcinoma (RCC) are the most common u... more Prostate cancer (PCa), bladder cancer (BCa), and renal cell carcinoma (RCC) are the most common urological cancers, and their incidence has been rising over time. Surgery is the standard treatment for these cancers, but this procedure is only effective when the disease is localized. For metastatic disease, PCa is typically treated with androgen deprivation therapy, while BCa is treated with chemotherapy, and RCC is managed primarily with targeted therapies. However, response rates to these therapeutic options remain unsatisfactory due to the development of resistance and treatment-related toxicity. Thus, the discovery of biomarkers with prognostic and predictive value is needed to stratify patients into different risk groups, minimizing overtreatment and the risk of drug resistance development. Pharmacometabolomics, a branch of metabolomics, is an attractive tool to predict drug response in an individual based on its own metabolic signature, which can be collected before, during, an...
This study aimed at investigating the impact of different technical cork stoppers on the quality ... more This study aimed at investigating the impact of different technical cork stoppers on the quality preservation and shelf life of sparkling wines. The volatile compositions of two Italian sparkling wines sealed with a sparkling cork with two natural cork discs (2D) and a microagglomerated (MA) cork were determined during bottle aging (12 to 42 months) after disgorging, by headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC/MS). The volatile profile of the sparkling wine #1 sealed with 2D stoppers showed the presence of camphor from 12 to 42 months, along with significant alterations in the levels of several alcohols, ketones, and ethyl esters at 24 and 42 months. The impact of closure type was less pronounced for sparkling wine #2 which also showed the presence of camphor from 12 to 42 months in 2D samples, and significantly higher levels of esters at 24 and 42 months for 2D compared with MA. These results unveiled that the type of closu...
The main purpose of this work was to describe and validate a new methodology for the determinatio... more The main purpose of this work was to describe and validate a new methodology for the determination of a brandy reactivity index (BRI) towards flavan-3-ol compounds in wines (catechins and condensed tannins) and to correlate this index with the amount and type of aldehydes present in those brandies. This new method consists of two main reactions: a) the ability of aldehydes to react with C8 and/or C6 positions in the phloroglucinol ring of a catechin present in excess (A ring); b) further reaction of the remaining free catechin with p-dimethylaminocinamaldehyde (DMACA) yielding to the formation of a blue compound that can be quantified by Visible spectroscopy at 640 nm. The impact of different experimental conditions such as reagent concentrations and reaction time on BRI were also evaluated. The method was validated through the determination of repeatability (intra-day variability) and reproducibility. The repeatability was considered acceptable with a CV of 11.87 %. The analysis of...
Adrenaline, which participates in the neuroendocrine response that occurs during stress and perim... more Adrenaline, which participates in the neuroendocrine response that occurs during stress and perimenopause, may be tumorigenic. This exploratory study aimed at investigating whether non-tumorigenic and tumorigenic human breast epithelial cell lines are able to synthesize adrenaline. The study was carried out in non-tumorigenic (MCF-10A) and tumorigenic (MCF-7) human breast cell lines. Expression of enzymes involved in adrenaline synthesis was characterized by RT-qPCR, immunocytochemistry and western blot. Catecholamines and analogue compounds were quantified by HPLC-ECD. Functional assessment of the impact of drugs on cells’ tumorigenic potential was assessed by determination of cell viability and clonogenic ability. Both MCF-10A and MCF-7 cells produce catecholamines, but the capacity to produce adrenaline is lower in MCF-10A cells. β-adrenoceptor activation increases the capacity of MCF-10A cells to produce adrenaline and favor both cell viability and colony formation. It is conclu...
Prostate cancer (PCa) is the second most diagnosed cancer in men worldwide. For its screening, se... more Prostate cancer (PCa) is the second most diagnosed cancer in men worldwide. For its screening, serum prostate specific antigen (PSA) test has been largely performed over the past decade, despite its lack of accuracy and inability to distinguish indolent from aggressive disease. Metabolomics has been widely applied in cancer biomarker discovery due to the well-known metabolic reprogramming characteristic of cancer cells. Most of the metabolomic studies have reported alterations in urine of PCa patients due its noninvasive collection, but the analysis of prostate tissue metabolome is an ideal approach to disclose specific modifications in PCa development. This review aims to summarize and discuss the most recent findings from tissue and urine metabolomic studies applied to PCa biomarker discovery. Eighteen metabolites were found consistently altered in PCa tissue among different studies, including alanine, arginine, uracil, glutamate, fumarate, and citrate. Urine metabolomic studies a...
An untargeted approach, focused in the profile of volatile organic compounds (VOCs), was applied ... more An untargeted approach, focused in the profile of volatile organic compounds (VOCs), was applied to differentiate natural cork stoppers with different levels of porosity, coded as Group 1 (low porosity), Group 2 (intermediate porosity) and Group 3 (high porosity). Statistically significant alterations were found in the levels of several VOCs between cork stoppers of low and intermediate porosity when compared with those of high porosity (Group 1 vs. 3 and Group 2 vs. 3). In addition, the levels of 2-pentylfuran, cyclene, camphene, camphor, limonene and eucalyptol enabled the discrimination of cork stoppers with low porosity (Group 1) into two subgroups, while furfural and 5-methyl-2-furfural enabled the discrimination of subgroups within the intermediate and high porosity stoppers (Group 2 and 3). A headspace solid-phase microextraction coupled to gas chromatography tandem mass spectrometry (HS-SPME-GC-MS/MS) method was developed to quantify the subgroup discriminant compounds, which may provide a proof-of-concept for the development of an efficient method to be applied in cork industry.
Clear cell renal cell carcinoma (ccRCC) is the most common type of kidney cancer usually associat... more Clear cell renal cell carcinoma (ccRCC) is the most common type of kidney cancer usually associated with asymptomatic development and risk of systemic progression. Hence, reliable molecular biomarkers of ccRCC are needed to provide early and minimally invasive detection. In this study, urinary volatilome profiling of patients diagnosed with ccRCC (n = 75), and cancer-free controls (n = 75), was performed to investigate the presence of a volatile signature characteristic of ccRCC. Volatile organic compounds (VOCs) in general, and more specifically volatile carbonyl compounds (VCCs), present in urine were extracted by headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS). Supervised multivariate models showed a good discriminatory power of ccRCC patients from controls in urine. Overall, 22 volatile metabolites were found significantly altered between the two groups, including aldehydes, ketones, aromatic hydrocarbons, and terpenoids. A candidate six-biomarker panel, comprising octanal, 3-methylbutanal, benzaldehyde, 2-furaldehyde, 4-heptanone, and p-cresol, depicted the best performance for ccRCC detection with 83% sensitivity, 79% specificity, and 81% accuracy. Moreover, the ccRCC urinary volatilome signature suggested dysregulation of energy metabolism and overexpression of enzymes associated with carcinogenesis. These findings provide the molecular basis for the fine-tuning of gas-sensing materials for application in the development of a bioelectronic sensor.
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