Percorrer por autor "Luís, Carla"
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- Anti-Angiogenic Properties of Cafestol and Kahweol Palmitate Diterpene EstersPublication . Moeenfard, Marzieh; Cortez, Alice; Machado, Vera; Costa, Raquel; Luís, Carla; Coelho, Pedro; Soares, Raquel; Alves, Arminda; Borges, Nuno; Santos, AlejandroEpidemiological studies support the association of coffee-specific diterpenes, with various beneficial health effects. Although anti-antiangiogenic properties of free cafestol and kahweol have been recently described, available data regarding their esterified form, in particular palmitate esters as the main diterpene esters present in coffee, are still rare. Given that angiogenesis plays an important role in many pathological conditions, including cancer growth and metastasis, this study aimed to assess and compare the potential anti-angiogenic effects of cafestol palmitate (CP) and kahweol palmitate (KP) in an in vitro angiogenesis model. According to our findings, both compounds inhibited angiogenesis steps on human microvascular endothelial cells (HMVECs), although a more significant effect was observed for KP. Compared to control, HMVECs viability decreased in a dose-dependent manner upon incubation either with CP or KP. Concentrations of 75 and 100 μM of each compound were cytotoxic. Cell proliferation was also dramatically reduced by both diterpene esters at 50 μM, although KP had a stronger inhibitory effect. However, CP and KP did not induce apoptosis on HMVECs. Both compounds reduced cell migration, but this effect was only statistically significant after KP incubation. Inhibition of VEGFR2 expression and its downstream effector Akt, but not Erk, was also observed in CP- and KP-treated HMVECs. These findings were confirmed using ELISA assay for phosphorylated (active) VEGFR-2. Taken together, these data indicate that both CP and KP can be considered potent compounds against angiogenesis-dependent disorders. Our findings further indicate that KP exerts more potent anti-angiogenic effects than CP, in most of assays.
- Avoiding the interference of doxorubicin with MTT measurements on the MCF-7 breast cancer cell linePublication . Luís, Carla; Castaño-Guerrero, Yuselis; Soares, Raquel; Sales, Goreti; Fernandes, RúbenDoxorubicin (DOXO) is an adjuvant chemotherapy agent and is also commonly used in cell biology research. Cytotoxic assays in cell culture are frequently used in order to stablish drug concentrations that are useful for controlling cell proliferation. One common cytotoxic method used is 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT). Our present research aims to support future studies in engaging MTT assay using DOXO that exhibits a strong red coloration and fluorescence, and so it is assumed that DOXO may interfere with commonly used colorimetric assays such as MTT. The interference of DOXO in the MTT determination was evaluated in a Breast Cancer cell line Michigan Cancer Foundation-7 (MCF-7). The interference was evaluated by means of spectroscopic methods in particular spectrophometry and fluorescence spectroscopy of MTT and DOXO. We postulate that the medium and the MTT reagent itself can interfere on the metabolic activity method, so in order to achieve better results, DMEM was replaced by a neutral buffer like Phosphate-buffered saline (PBS). This protocol may be extremely useful in future studies involving DOXO.
- Bilateral breast cancer and the influence of body mass index in clinicopathological features and overall survivalPublication . Luís, Carla; Fernandes, Rute; Dias, João; Pereira, Deolinda; Firmino-Machado, João; Baylina, Pilar; Fernandes, Rúben; Soares, Raquel; BAYLINA MACHADO, PILARBreast cancer (BC) and obesity are two closely associated pathologies with increasing incidence and mortality rates. Bilateral Breast Cancer (BBC) displays a low incidence rate within BC and obesity represents a major risk factor. The aim of this study is to analyzed BBC clinicopathological features distribution and determine the potential influence of obesity in BBC in these same features and overall survival. Clinicopathological information was obtained from 42 cases of women with BBC diagnosed in IPO-Porto. To evaluate the frequency distribution of the clinicopathological data, a chi-square goodness of fit test was performed for BBC cases. A chi-square test of independence was applied for BMI stratification. Cox regression was performed for overall survival. Statistical significance was set at p-value < 0.05. Distribution of BBC clinicopathological features was found to be statistically significant in family history (p-value < 0.001), BBC type (p-value < 0.001), stage (p-value = 0.005), differentiation grade (p-value < 0.001), receptor expression (pvalue < 0.001) and histological type (p-value = 0.031). In comparison to the statistical expected results, we observed an increased cases of absence of family history and less cases of metachronous BBC. Histological types between tumours of BBC were mostly concordant. All cases presented concordant receptor expression. Analysis stratified by BMI revealed that obese women were diagnosed later, although without statistical significance. All obese women presented poor differentiation grade (n = 6). Overweight patients display a tendency to a better overall survival with lower tumour stages and lower differentiation grades. Our results reveal the same receptor expression between contralateral tumours. Also, most tumours share the same histological type. When stratified by BMI, we observed a tendency for overweight women to have improved overall survival.
- Characterization of carbon dots and its impact in a breast cancer cell linePublication . Magalhães, Sofia; Luís, Carla; Fraga, Hugo; Duarte, Abel J.Carbon dots (CD) have emerged as a novel promising alternative to metal-based nanoparticles. These fluorescence carbon materials have been the subject of research since 2004 and exhibit excellent chemical and physical properties such as hydrophilicity, good biocompatibility and reduced toxicity, which allows a good interaction with biological systems. Taking this in consideration it is not surprising that CD have been introduced for the study/therapeutics of breast cancer , especially, in drug delivery system, bioimaging, biosensor, microbial therapy, photodynamic and photothermal therapy. Breast cancer is the most prevalent cancer with a high morbidity rate in the female population worldwide. In this study, we considered the use of fructose derived CD for breast cancer therapy. CD were produced from D-fructose via microwave irradiation by the bottom-up method. Following purification, particles were characterized using TEM, FTIR, DLS and fluorescence microscopy. As expected, fructose derived CD consist of carbon, oxygen and hydrogen. As characteristic of CD, they are fluorescent with an absorption maximum at 360 nm that when irradiated at 360 nm have an emission maximum at 520 and a broad emission band (maximum 520 nm). In what concern cell-based assays, our results using breast cancer cell line BT474 and MCF-10A breast epithelial cells show CD are able to penetrate the cell membrane and display cytotoxicity towards tumor cell lines. In still preliminary results, it appears that they have reduced toxicity versus non tumor cell line, suggesting that they can be used as anti-tumoral agents for therapeutic purposes.
- Exploring the anti-cancer properties of pomegranate peel aqueous extractPublication . Luís, Carla; Sousa, André P.; Costa, Raquel; Maduro, Ana T.; Pais, Patrick J.; Sá, Sara; Gestoso, Álvaro; Fernandes, Flávia; Jerónimo, Eliana; Soares, Raquel; Fernandes, Rúben; Baylina, Pilar; Duarte, Maria F.The objective of this work is to evaluate the influence of pomegranate peel extract (PPE) in the behavior of breast cell lines (epithelial and tumor type) and related oxidative metabolism. Fruitbased functional foods have been the target of increasing scientific research for their physiological and pathophysiological properties. Pomegranate (Punica granatum) is a suitable example with both prophylactic and medicinal effects. MCF-7 cell line from tumor breast carcinoma, and MCF-10A cell line from normal epithelial mammary gland were used and subjected to different concentrations of PPE, ranging from 1 to 5 mM of gallic acid equivalents (GAE). Viability, proliferation, mobility, and cytotoxicity assays were performed along with the quantification of antioxidant enzymes, namely, catalase, superoxide dismutase (SOD) and reduced (GSH) and oxidized (GSSG) glutathione. We observed a decrease in viability and proliferation of MCF-7 cells, at higher concentrations of PPE, with no influence in epithelial cells. Interestingly, in a concentration-dependent manner, PPE triggered a significant decrease in migration on both cell lines, with a more pronounced effect in breast cancer cell line. Regarding antioxidant enzyme activity, on tumor cells higher concentrations of PPE decreased catalase activity and significantly increased SOD activity. Regarding GSH and GSSG, we observed different expression levels between MCF-7 and MCF-10A, with MCF-7 presenting lower levels compared to MCF-10A. GSH/GSSG ratio was notably higher in MCF-7 at 5 mM GAE. PPE exhibits anti-tumor effects without significantly affecting normal epithelial cells. Our work strengthens the potential antitumoral effect of PPE by reducing MCF-7 cell viability and proliferation through the imbalance of antioxidant enzymes.
- Fructose-Derived Carbon Dots as Selective Antitumor Agents in Breast Cancer Therapy: Synthesis, Characterization, and Biological EvaluationPublication . Magalhães, Sofia; Luís, Carla; Duarte, Abel J.; Adams, James DavidThis study explored a novel method using fructose-derived carbon dots (FCDs) for antitumor therapy in breast cancer (BC), marking a pioneering use of fructose as a carbon source for nanoparticle synthesis. BC, known for its complexity and heterogeneity, was chosen as a model due to its increasing mortality and incidence rates. The FCD synthesis involved the decomposition of fructose through microwave irradiation, followed by purification and characterization using techniques such as transmission electron microscopy, dynamic light scattering, fluorescence spectrophotometry, and Fourier-transform infrared spectroscopy. The FCDs, ranging in size from 2 to 6 nm, presented a hydrodynamic diameter below 2 nm, a spherical morphology, and a crystalline structure. As expected, FCDs were composed by carbon, oxygen, and hydrogen, and exhibited fluorescence with absorption and emission spectra at 405 nm and around 520 nm, respectively. Cell-based assays on breast epithelial and tumor cell lines demonstrated a dose-dependent response, with a decreased viability rate more pronounced in breast tumor cells. In conclusion, FCDs showed significant potential as selective antitumor agents for breast cancer therapy. The comprehensive characterization and cell-based assay evaluations provided valuable insights into the applications of these nanoparticles in breast cancer treatment, highlighting their selective toxicity and impact on tumor cells.
- GLP-1R and IL-6 expression in the gastrointestinal tract of a murine model of metabolic syndromePublication . Costa, José; Oliveira, Sofia; Pereira, Ana Cláudia; Soares, Raquel; Baylina, Pilar; Guerreiro, Susana G.; Fernandes, Rúben; Luís, CarlaGlucose homeostasis is a critical cornerstone in both health and disease. It is described to be regulated by the balance of insulin and glucagon secretion but, this bi-hormonal perspective is long overdue, since glucose homeostasis is now known to be a multi-hormonal process. Metabolic syndrome (MetS) is a cluster of metabolic features which includes impaired glucose metabolism and obesity. Obesity promotes a low chronic inflammation state due to release of bioactive molecules like cytokines, ultimately contributing to cardiovascular disease, diabetes and cancer. The gastrointestinal tract (GIT), namely stomach and intestine, has a vital role not only in the food uptake and absorption, but also in the production of incretin hormones, such as GLP-1. We aim to evaluate the GLP-1 receptor (GLP-1R) in the GIT of a MetS animal model, and to assess whether it correlates with inflammatory levels. The expression of GLP-1R and Interleukin-6 (IL-6) was evaluated in the stomach and intestine of mice subjected to Normal Diet (ND) and High Fat Diet (HFD) by immunohistochemistry. We observed that HFD fed animals presented lower levels of GLP-1R in stomach and intestine when compared with animals fed with ND. Concomitantly, these mice expressed increased levels of IL-6. GLP-1R expression is inversely correlated with the expression of the proinflammatory cytokine IL-6.
- Harvesting the power of green synthesis: gold nanoparticles tailored for prostate cancer therapyPublication . Oliveira, Marco; Sousa, André; Sá, Sara; Soares, Sílvia; Pereira, Ana Cláudia; Rocha, Ana Catarina; Pais, Patrick; Ferreira, Diogo; Almeida, Cátia; Luís, Carla; Lima, Cláudio; Almeida, Fábio; Gestoso, Álvaro; Duarte, Miguel-Correa; Barata, Pedro; Martins-Mendes, Daniela; Baylina, Pilar; Pereira, Carla F.; Fernandes, RúbenBiosynthetic gold nanoparticles (bAuNPs) present a promising avenue for enhancing biocompatibility and offering an economically and environmentally responsible alternative to traditional production methods, achieved through a reduction in the use of hazardous chemicals. While the potential of bAuNPs as anticancer agents has been explored, there is a limited body of research focusing on the crucial physicochemical conditions influencing bAuNP production. In this study, we aim to identify the optimal growth phase of Pseudomonas aeruginosa cultures that maximizes the redox potential and coordinates the formation of bAuNPs with increased efficiency. The investigation employs 2,6-dichlorophenolindophenol (DCIP) as a redox indicator. Simultaneously, we explore the impact of temperature, pH, and incubation duration on the biosynthesis of bAuNPs, with a specific emphasis on their potential application as antitumor agents. Characterization of the resulting bAuNPs is conducted using ATR-FT-IR, TEM, and UV-Vis spectroscopy. To gain insights into the anticancer potential of bAuNPs, an experimental model is employed, utilizing both non-neoplastic (HPEpiC) and neoplastic (PC3) epithelial cell lines. Notably, P. aeruginosa cultures at 9 h/OD600 = 1, combined with biosynthesis at pH 9.0 for 24 h at 58 ◦C, produce bAuNPs that exhibit smaller, more spherical, and less aggregated characteristics. Crucially, these nanoparticles demonstrate negligible effects on HPEpiC cells while significantly impacting PC3 cells, resulting in reduced viability, migration, and lower IL-6 levels. This research lays the groundwork for the development of more specialized, economical, and ecologically friendly treatment modalities.
- The implementation of HFMEA in a primary care emergency service of the portuguese national health systemPublication . Macedo, José Mário; Sá, Sara; Gestoso, Álvaro; Ramalho, Susana; Luís, Carla; Martins-Mendes, Daniela; Pereira, Ana Cláudia; Fernandes, Rúben; Baylina, Pilar; BAYLINA MACHADO, PILAROne of the main priorities for healthcare institutions is to provide better and more advanced services that contribute to prolonging the lives of those who seek their care, as well as improving their quality of life as long as it lasts. Both their patients and the professionals who work in those institutions need to feel protected against adverse effects that may affect them physically, mentally, or socially. As such, both at national and international levels, healthcare governing bodies have made decisions and implemented measures aimed at reducing the risk of adverse effects for the patients who use their services. The main objective of this work was to develop and implement a risk management system based on HFMEA in a basic emergency service (primary care), to promote patient safety and the safety of healthcare professionals. A prospective, explanatory, and descriptive case study was applied to a basic emergency service, using ISO 31000 – risk management in organisations, and the healthcare failure mode and effects analyses (HFMEA) support tool. There were identified 57 possible failures associated with 88 different effects, with risk magnitudes classified as severe (1 cases), very severe (18 cases), and intolerable (38 cases). Failures whose effects impact patients were mentioned 2,6 times more than those primarily affecting professionals. The study allowed the identification of different failures in some of its work processes, for which corrective measures were determined to be carried out by the team and the governing entities of the service, to be implemented as soon as possible, ensuring greater safety in its services.
- Low skeletal muscle function, but not mass, is associated with the presence of type 2 DiabetesPublication . Rigor, Joana; Barbosa, João Portugal; Luís, Carla; Fernandes, Rúben; Barata, Pedro; Martins-Mendes, DanielaThe pathophysiology of type 2 Diabetes mellitus (T2DM) is intimately connected to the skeletal muscle (SkM). SkM affects insulin resistance and is, in turn, affected by the metainflammation, microvascular disease and ectopic fat deposition of T2DM. SkM mass can be inferred by the waist-to-calf ratio (WCR) and its function by the Short Physical Performance Battery (SPPB). The aim of this study was to determine the association between SkM mass and function with T2DM in patients with Metabolic Syndrome (MetS). Patients with MetS, aged 18 to 75 years-old, attending an outpatient clinic from April 15th to September 30th 2019, were consecutively included. Exclusion criteria comprised type 1 Diabetes, secondary hypertension, active neoplasia, autoimmune disease, HIV or hepatitis virus B or C infection and end-stage renal disease and/or liver disease. History and anthropometric data were collected, including weight, height, waist circumference (WC) and WCR; the SPPB was applied. A total of 81 patients were included, of which 58.0% had T2DM; most patients were female (55.6%) and the median age was 65 (interquartile range 16.5) years. Patients with T2DM were older (64.1 vs. 56.5 years, p=0.001) and more likely to have concurrent hypertension (96% vs. 65%, p<0.001) and dyslipidemia (96% vs. 56%, p<0.001). In univariate analysis, WC [odds ratio (OR) 1.1, 95% confidence interval (CI) 1.0-1.1), WCR (OR 146.2, 95% CI 9.9-2159.0) and SPPB (0.6, 95% CI 0.4-0.8) were associated with T2DM. In multivariate analysis, only SPPB maintained its association (OR 0.65, 95% CI 0.44-0.97). Poorer muscle function, as determined by the SPPB, was associated with the presence of T2DM, even when considering body composition, per WCR. Longitudinal and mechanistic studies are warranted to best characterize this relationship.
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