Search

Prostate cancer (PCa) is a global health problem that affects millions of men every year. In the past decade, metabolomics and related subareas, such as lipidomics, have demonstrated an enormous potential to identify novel mechanisms underlying PCa development and progression, providing a good basis for the development of new and more effective therapies and diagnostics. In this study, a multiplatform metabolomics and lipidomics approach, combining untargeted mass spectrometry (MS) and nuclear magnetic resonance (NMR)-based techniques, was applied to PCa tissues to investigate dysregulations associated with PCa development, in a cohort of 40 patients submitted to radical prostatectomy for PCa. Results revealed significant alterations in the levels of 26 metabolites and 21 phospholipid species in PCa tissue compared with adjacent nonmalignant tissue, suggesting dysregulation in 13 metabolic pathways associated with PCa development. The most affected metabolic pathways were amino acid metabolism, nicotinate and nicotinamide metabolism, purine metabolism, and glycerophospholipid metabolism. A clear interconnection between metabolites and phospholipid species participating in these pathways was observed through correlation analysis. Overall, these dysregulations may reflect the reprogramming of metabolic responses to produce high levels of cellular building blocks required for rapid PCa cell proliferation.

Background: QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) is a methodology previously developed to extract pesticides from vegetables and fruits and has been fully applied for different analytical approaches. Objective: In the present study, a rapid and less laborious modified QuEChERS extraction method for the quantification of 13 opioids [codeine, morphine, heroin, 6-acetylmorphine (6-AM), desomorphine, ethylmorphine, methadone, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), 2-ethyl-5-methyl-3,3- diphenyl-1-pyrroline (EMDP), papaverine, tramadol, O-desmetyltramadol (M1) and, tapentadol], cocaine and cocaethylene in whole blood was developed and validated by Gas Chromatography-Mass Spectrometry. Method: The modification of QuEChERS method consisted in the pretreatment of the whole blood samples using ultrasonication, the use of ethyl acetate as extraction solvent and a previous step of sample alcalinization. The use of dispersive separation steps such as Dispersive Solid-Phase Extraction (dSPE) or sorbents such as Primary Secondary Amine (PSA) was suppressed to minimize the errors and, to improve the velocity of the analysis. Results: The method proved to be selective and the regression analysis for the analytes was linear in the range of 31.2-2000 ng/mL with correlation coefficients > 0.98. The coefficients of variation did not exceed 15%. The lowest limit of detection and quantification for all the analytes were below the therapeutic range of the drugs. The recoveries of the analytes ranged from 52.4 to 95.0%. Conclusion: The developed method can provide a rapid, effective and “greener” process for the analysis of a wide range of opioids drugs in whole blood samples and can be applied to clinical and forensic antemortem and postmortem cases.

Prostate cancer (PCa) is the most frequently diagnosed cancer and the second leading cause of cancer death among men in Western countries. Current screening techniques are based on the measurement of serum prostate specific antigen (PSA) levels and digital rectal examination. A decisive diagnosis of PCa is based on prostate biopsies; however, this approach can lead to false-positive and false-negative results. Therefore, it is important to discover new biomarkers for the diagnosis of PCa, preferably noninvasive ones. Metabolomics is an approach that allows the analysis of the entire metabolic profile of a biological system. As neoplastic cells have a unique metabolic phenotype related to cancer development and progression, the identification of dysfunctional metabolic pathways using metabolomics can be used to discover cancer biomarkers and therapeutic targets. In this study, we review several metabolomics studies performed in prostatic fluid, blood plasma/serum, urine, tissues and immortalized cultured cell lines with the objective of discovering alterations in the metabolic phenotype of PCa and thus discovering new biomarkers for the diagnosis of PCa. Encouraging results using metabolomics have been reported for PCa, with sarcosine being one of the most promising biomarkers identified to date. However, the use of sarcosine as a PCa biomarker in the clinic remains a controversial issue within the scientific community. Beyond sarcosine, other metabolites are considered to be biomarkers for PCa, but they still need clinical validation. Despite the lack of metabolomics biomarkers reaching clinical practice, metabolomics proved to be a powerful tool in the discovery of new biomarkers for PCa detection.

The incorporation of lysine acetylsalicylate (LAS) in the commercial formulation of paraquat (PQ), Gramoxone®, has been shown to significantly increase the survival of intoxicated mammals while maintaining the herbicidal effect. The aim of the present study was to clarify the mechanisms involved in the protective effect of LAS in a rodent model through monitoring PQ levels and the histological and biochemical biomarkers of toxicity in male Wistar rats following intoxication with PQ formulations in the absence and presence of LAS. Gramoxone® and the formulation with LAS were administered by gavage at the doses of 125 mg kg−1of PQ and 125 mg kg−1of PQ + 316 mg kg−1of LAS, respectively. The obtained results showed that LAS improves the safety of PQ formulation by increasing its elimination and preventing lung and kidney injury. LAS prevented the biochemical and histological alterations in lung induced by PQ measured at the end of 24 h and 48 h. This was evidenced by a significant reduction in lipid peroxidation, the maintenance of reduced glutathione levels and decreased levels of oxidized glutathione, as well as the normalization of the urinary biomarkers, creatinine and N-acetyl-β-glucosaminidase. LAS treatment also caused a significant reduction in PQ-induced activation of nuclear factor kappa B (NF-κB) in the lung. The results allow us to conclude that lysine acetylsalicylate improves the safety of PQ formulation in rats by increasing its elimination and preventing lung and kidney injury.

Remarkable interindividual differences in 3,4-methylenedioxymethamphetamine (‘Ecstasy’)-mediated toxicity have been reported in humans. Therefore, we tested whether CYP2D6 or its variant alleles as well as CYP3A4 influence the susceptibility to 3,4-methylenedioxymethamphetamine.

The consumption of amphetamine is illicit and controlled due to both the elicited behavioural deviations and the toxicity effects reported in abusers. Thus, amphetamine levels in biological samples must be monitored in several clinical and forensic circumstances. In spite of the interspecies differences in the preferred route of biotransformation, benzylmethylketone, benzoic acid and 4-hydroxyamphetamine are the principal metabolites of amphetamine. However, the clinical and forensic studies are focused in the parent compound and in 4-hydroxyamphetamine since benzylmethylketone is a minor metabolite in human and benzoic acid is also an endogenous compound. In the present study amphetamine and its metabolite, 4-hydroxyamphetamine, are quantified in urine by HPLC after derivatization with 4-dimethylaminoazobenzene-4'-sulfonyl chloride (dabsyl chloride). This derivatization procedure transforms amphetamine and its hydroxylated metabolite in compounds with similar lipofilicity, enabling their quantitative and simultaneous extraction with an organic solvent. The precision of the HPLC technique was 7.3 and 10.0% for amphetamine and 4-hydroxyamphetamine derivatives, respectively. For the overall procedure, including enzymatic hydrolysis, derivatization and extraction of the derivatives, the obtained values were 9.3 and 6.2%. Recoveries obtained from spiked urines for amphetamine and 4-hydroxyamphetamine were better than 97% and 94% (mean value), respectively. The detection limits of the method was 10 ng for both compounds. The principal advantages of the present proposed method are the stability of the dabsyl derivatives at room temperature and the detection carried out in the visible region, reducing the interferences detected. Copyright © 2001 John Wiley & Sons, Ltd.

A comparative study to extract and purify total morphine in urine is described using several procedure treatments of biological samples. Sep-Pak C18 cartridges, Extrelut columns and liquid-liquid conventional extraction with sequencial purification through Extrelut column were assayed in order to establish the best procedure to quantitatively extract morphine from urine free of endogenous interferents.Analysis of the drug was made by HPLC-UV with a C18 column and methanol: acetate buffer (pH6.9) (70+30) as eluent.Recovery of morphine from spiked urine was 88.0% and 79.8% for concentrations of 1μg/ml and 5μg/ml, respectively. The detection limit of the method was 0.2μg/ml and coeficient of variation was 1.8% (n+4).

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.