O efeito dos extractos de cereja em células humanas do cancro da próstata: Do cultivo à clínica?

A proliferação descontrolada das células da próstata é uma condição observada normalmente na população masculina à medida que esta envelhece, e está associada com o desenvolvimento do cancro da próstata. Assim, estratégias que visem diminuir a proliferação dos tecidos prostáticos durante o seu envelhecimento deverão diminuir a prevalêcia desta condição médica. As cerejas são ricas em compostos fenólicos como as antocianinas, com efeitos antioxidantes e anti-proliferativos bem estudados em células cancerígenas como as MCF-7 (cancro da mama) e HCT116 (cancro colo-rectal). O estudo elaborado para esta dissertação pretendeu avaliar o efeito de extractos da cereja Saco, uma variedade local da região do Fundão, na viabilidade, apoptose, e metabolismo glicolítico das células não-neoplásicas do epitélio da próstata PNT1A, nas células neoplásicas androgeno-dependentes LNCap, e nas células neoplásicas androgeno-independentes PC3. As cerejas foram descaroçadas, liofilizadas, e a sua extracção efectuada recorrendo a ultrassons e utilizando metanol acidificado com 0,1% HCl como solvente. O extracto obtido foi então dissolvido em meio de cultura celular a várias concentrações (0, 2, 20, 200 e 2000 µg/mL) e utilizado para tratar as células não-neoplásicas e neoplásicas durante 48, 72 e 96 horas. A viabilidade celular nas diferentes condições experimentais foi determinada por ensaios MTT. Globalmente, a viabilidade celular mais baixa foi observada às 72 horas e com 20 µg/mL de extracto de cereja, e estas condições foram utilizadas para avaliar os efeitos dos extractos de cereja na proliferação,apoptose e metabolismo glicolítico das células PNT1A, LNCaP e PC3. A análise à expressão génica e a actividade dos reguladores-alvo destes processos biológicos foram efectuadas através da metodologia de Western Blots e kits de análises bioquimicas, respectivamente. O consumo de glucose e a produção de lactato foram ambos medidos por análise espectofotométrica. Os resultados obtidos demonstraram que a administração dos extractos metanólicos de cereja aumentou a apoptose das células LNCaP, um efeito não-confirmado nas células não-neoplásicas PNT1A. Curiosamente, as células PC3 mostraram uma diminuição da actividade apoptótica na presença dos extractos de cereja, apesar da proliferação diminuída. Quanto ao metabolismo glicolítico, foi observado que as células PNT1A tratadas com extractos de cereja apresentaram uma diminuição parcial deste, enquanto que as células LNCaP demonstraram uma supressão da metabolização da glucose em resposta aos extractos de cereja, com consumo de glucose e produção de lactato ambos diminuídos, acompanhados por alterações da expressão/actividade de transportadores e enzimas glicolíticas. Analogamente aos resultados da apoptose, as células PC3 apresentaram um comportamento distinto das LNCaP relativamente ao metabolismo glicolítico; os extractos de cereja induziram um aumento da glicólise e da produção de lactato. Os presentes resultados indicam que os extractos metanólicos de cereja podem ter efeitos benéficos nas células do cancro da próstata, tanto neoplásicas como não-neoplásicas, diminuindo a sua proliferação e controlando as vias apoptóticas e glicolíticas, produzindo efeitos mais visíveis no caso das células sensíveis à ação dos androgénios, LNCaP.The uncontrolled proliferation of prostate cells is a condition commonly observed in the ageing male population, which is associated with the development of prostate cancer. Thus, strategies to counteract the proliferative feature of aging prostatic tissues should prove beneficial by diminishing the widespread prevalence of this condition. Sweet cherries are rich in phenolic compounds such as anthocyanins, with widely studied anti-proliferative and antioxidant effects in different types of cancer cells such as MCF-7 (human breast) and HCT116 (human colorectal). The present study aimed to evaluate the effect of crude extracts of Saco sweet cherry, a local cultivar from the Fundão region (Portugal), on the viability, apoptosis and glycolytic metabolism of non-neoplastic PNT1A prostate epithelial cells, and androgen-sensitive LNCaP cells and androgen-insensitive PC3 prostate cancer cells. Sweet cherries were seeded, freeze-dried, and underwent ultrasound-assisted extraction with methanol acidified with HCl 0.1%. The extract was then dissolved in cell culture medium at several concentrations (0, 2, 20, 200 and 2000 µg/mL) and used to treat non-neoplastic and neoplastic prostate cells for 48, 72 and 96 hours. Cell viability at different experimental conditions was determined by MTT assay. Overall, the lowest viability was observed at 72 hours of culture with 20 µg/mL of cherry extract, and these conditions were used to evaluate the effect of cherry extracts on the cell proliferation, apoptosis and glycolytic metabolism of PNT1A, LNCaP and PC3 cells. Gene expression analysis and activity of target regulators of the aforementioned biological processes was assessed by means of Western blot and biochemical assays, respectively. Glucose consumption and lactate production were measured spectrophotometrically. The obtained results demonstrated that the administration of methanolic cherry extracts increased the apoptotic rate of LNCaP cells, an effect not confirmed in the non-neoplastic PNT1A cells. Curiously, PC3 cells had an overall down-regulation of apoptotic activity in the presence of cherry extracts, in spite of its decreased proliferation. Concerning metabolism, it was found that treated PNT1A cells had a partial down-regulation of glycolytic metabolism, while LNCaP cells markedly displayed a suppression of this process with decreased glucose consumption and lactate production, which was accompanied by altered expression/activity of glycolytic transporters and enzymes. Analogously with the results of apoptosis, PC3 cells had a distinct behavior relatively to LNCaP in what concerns glycolytic metabolism; cherry extracts induced an up-regulation of glycolysis and lactate production. The present findings indicate that cherry methanolic extracts may have beneficial effects on prostate cells, both neoplastic and non-neoplastic, diminishing proliferation and controlling the apoptotic and glycolytic pathways, producing more visible effects in the case of the androgen-sensitive LNCaP cells

ReproducibiliTea Cup at QUB

Materials from ReproducibiliTea sessions in Cup at QUB. Templates and presentations are available for others to use and edit

Identification of milk from different animal and plant sources by desorption electrospray ionisation high-resolution mass spectrometry (DESI-MS)

This study used desorption electrospray ionisation mass spectrometry (DESI-MS) to analyse and detect and classify biomarkers in five different animal and plant sources of milk for the first time. A range of differences in terms of features was observed in the spectra of cow milk, goat milk, camel milk, soya milk, and oat milk. Chemometric modelling was then used to classify the mass spectra data, enabling unique or significant markers for each milk source to be identified. The classification of different milk sources was achieved with a cross-validation percentage rate of 100% through linear discriminate analysis (LDA) with high sensitivity to adulteration (0.1–5% v/v). The DESI-MS results from the milk samples analysed show the methodology to have high classification accuracy, and in the absence of complex sample clean-up which is often associated with authenticity testing, to be a rapid and efficient approach for milk fraud control

The General Growth Tendency: A tool to improve publication trend reporting by removing record inflation bias and enabling quantitative trend analysis

The trend of the number of publications on a research field is often used to quantify research interest and effort, but this measure is biased by general publication record inflation. This study introduces a novel metric as an unbiased and quantitative tool for trend analysis and bibliometrics. The metric was used to reanalyze reported publication trends and perform in-depth trend analyses on patent groups and a broad range of field in the life-sciences. The analyses confirmed that inflation bias frequently results in the incorrect identification of field-specific increased growth. It was shown that the metric enables a more detailed, quantitative and robust trend analysis of peer reviewed publications and patents. Some examples of the metric’s uses are quantifying inflation-corrected growth in research regarding microplastics (51% ± 10%) between 2012 and 2018 and detecting inflation-corrected growth increase for transcriptomics and metabolomics compared to genomics and proteomics (Tukey post hoc p&lt;0.0001). The developed trend-analysis tool removes inflation bias from bibliometric trend analyses. The metric improves evidence-driven decision-making regarding research effort investment and funding allocation.</p

Data fusion and multivariate analysis for food authenticity analysis

Abstract A mid-level data fusion coupled with multivariate analysis approach is applied to dual-platform mass spectrometry data sets using Rapid Evaporative Ionization Mass Spectrometry and Inductively Coupled Plasma Mass Spectrometry to determine the correct classification of salmon origin and production methods. Salmon (n = 522) from five different regions and two production methods are used in the study. The method achieves a cross-validation classification accuracy of 100% and all test samples (n = 17) have their origins correctly determined, which is not possible with single-platform methods. Eighteen robust lipid markers and nine elemental markers are found, which provide robust evidence of the provenance of the salmon. Thus, we demonstrate that our mid-level data fusion - multivariate analysis strategy greatly improves the ability to correctly identify the geographical origin and production method of salmon, and this innovative approach can be applied to many other food authenticity applications