Non-targeted LC-MS based metabolomics analysis of the urinary steroidal profile

The urinary steroidal fraction has been extensively explored as non-invasive alternative to monitor pathological conditions as well as to unveil the illicit intake of pseudo-endogenous anabolic steroids in sport. However, the majority of previous approaches involved the a priori selection of potentially relevant target analytes. Here we describe the non-targeted analysis of the urinary steroidal profiles. The workflow includes minimal sample pretreatment and normalization according to the specific gravity of urine, a 20 min reverse phase ultra-performance liquid chromatographic separation hyphenated to electrospray time-of-flight mass spectrometry. As initial validation, we analyzed a set of quality control urines spiked with glucurono- and sulfo-conjugated steroids at physiological ranges. We then applied the method for the analysis of samples collected after single transdermal administration of testosterone in hypogonadal men. The method allowed profiling of approximately three thousand metabolic features, including steroids of clinical and forensic relevance. It successfully identified metabolic pathways mostly responsible for groups clustering even in the context of high inter-individual variability and allowed the detection of currently unknown metabolic features correlating with testosterone administration. These outcomes set the stage for future studies aimed at implementing currently monitored urinary steroidal markers both in clinical and forensic analysis

The use of cytochrome P450 inhibitors in sport. A new generation of doping masking agents?

The activity of the CYP450 enzymes responsible for the phase I metabolism of most of the compounds included in the World Anti-Doping Agency (WADA) list of prohibited substances and methods could be strongly modified by the combined administration of other drugs such as, for example, the antidepressant, the antifungal and the H2 receptor antagonist agents. These compounds act as inhibitors of the CYP450 isoforms and it has been demonstrated that their co-administration with a drug that is also a CYP450 substrate may lead to a substantial alteration of the latter drug bioavailability, metabolism and excretion kinetics. In sports some classes of non-banned drugs, and primarily among them antidepressants, antifungals and the H2 receptor antagonists are extensively used, according to the information available on the doping control forms. Athletes may intentionally combine the CYP450 inhibitors with doping agents to modify in urine the time window of detection of the selected marker(s) of drug abuse, especially in those cases where the parent drugs are extensively metabolized

Editorial: OMICS-based approaches in sports research volume II.

OMICS approaches, including genomics, epigenomics, transcriptomics, proteomics, and metabolomics, continue to provide invaluable tools for better understanding of the molecular mechanisms underlying various physiological and pathological functions in health and disease. The rapid advancement of these tools and emergence of new ones is progressively filling the gap in our understanding of the complex networks that determine the structure, function, and dynamics of organisms. These advancements have greatly empowered discoveries of novel diagnostic and prognostic biomarkers as well as therapeutic targets, and provided a better guidance to precision and personalized medicine

Thu0349 autologous fat grafting in the treatment of patients with systemic sclerosis: current experience and future prospects

none13nomixedSpinella, Amelia; Pignatti, Marco; Citriniti, Giorgia; Lumetti, Federica; Cocchiara, Emanuele; Palermo, Adalgisa; Sighinolfi, Gianluca; Pacchioni, Lucrezia; Zaccaria, Giovanna; Lusetti, Irene Laura; Santis, Giorgio De; Salvarani, Carlo; Giuggioli, DiliaSpinella, Amelia; Pignatti, Marco; Citriniti, Giorgia; Lumetti, Federica; Cocchiara, Emanuele; Palermo, Adalgisa; Sighinolfi, Gianluca; Pacchioni, Lucrezia; Zaccaria, Giovanna; Lusetti, Irene Laura; Santis, Giorgio De; Salvarani, Carlo; Giuggioli, Dili

Palbociclib and fulvestrant act in synergy to modulate central carbon metabolism in breast cancer cells

The aims of this study were to determine whether combination chemotherapeutics exhibit a synergistic effect on breast cancer cell metabolism. Palbociclib, is a selective inhibitor of cyclin-dependent kinases 4 and 6, and when patients are treated in combination with fulvestrant, an estrogen receptor antagonist, they have improved progression-free survival. The mechanisms for this survival advantage are not known. Therefore, we analyzed metabolic and transcriptomic changes in MCF-7 cells following single and combination chemotherapy to determine whether selective metabolic pathways are targeted during these different modes of treatment. Individually, the drugs caused metabolic disruption to the same metabolic pathways, however fulvestrant additionally attenuated the pentose phosphate pathway and the production of important coenzymes. A comprehensive effect was observed when the drugs were applied together, confirming the combinatory therapy’s synergism in the cell model. This study also highlights the power of merging high-dimensional datasets to unravel mechanisms involved in cancer metabolism and therapy

Effect of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker initiation on organ support-free days in patients hospitalized with COVID-19

IMPORTANCE Overactivation of the renin-angiotensin system (RAS) may contribute to poor clinical outcomes in patients with COVID-19. Objective To determine whether angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) initiation improves outcomes in patients hospitalized for COVID-19. DESIGN, SETTING, AND PARTICIPANTS In an ongoing, adaptive platform randomized clinical trial, 721 critically ill and 58 non–critically ill hospitalized adults were randomized to receive an RAS inhibitor or control between March 16, 2021, and February 25, 2022, at 69 sites in 7 countries (final follow-up on June 1, 2022). INTERVENTIONS Patients were randomized to receive open-label initiation of an ACE inhibitor (n = 257), ARB (n = 248), ARB in combination with DMX-200 (a chemokine receptor-2 inhibitor; n = 10), or no RAS inhibitor (control; n = 264) for up to 10 days. MAIN OUTCOMES AND MEASURES The primary outcome was organ support–free days, a composite of hospital survival and days alive without cardiovascular or respiratory organ support through 21 days. The primary analysis was a bayesian cumulative logistic model. Odds ratios (ORs) greater than 1 represent improved outcomes. RESULTS On February 25, 2022, enrollment was discontinued due to safety concerns. Among 679 critically ill patients with available primary outcome data, the median age was 56 years and 239 participants (35.2%) were women. Median (IQR) organ support–free days among critically ill patients was 10 (–1 to 16) in the ACE inhibitor group (n = 231), 8 (–1 to 17) in the ARB group (n = 217), and 12 (0 to 17) in the control group (n = 231) (median adjusted odds ratios of 0.77 [95% bayesian credible interval, 0.58-1.06] for improvement for ACE inhibitor and 0.76 [95% credible interval, 0.56-1.05] for ARB compared with control). The posterior probabilities that ACE inhibitors and ARBs worsened organ support–free days compared with control were 94.9% and 95.4%, respectively. Hospital survival occurred in 166 of 231 critically ill participants (71.9%) in the ACE inhibitor group, 152 of 217 (70.0%) in the ARB group, and 182 of 231 (78.8%) in the control group (posterior probabilities that ACE inhibitor and ARB worsened hospital survival compared with control were 95.3% and 98.1%, respectively). CONCLUSIONS AND RELEVANCE In this trial, among critically ill adults with COVID-19, initiation of an ACE inhibitor or ARB did not improve, and likely worsened, clinical outcomes. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0273570

Editorial: OMICS-Based Approaches in Sports Research.

Genomics, transcriptomics and metabolomics are increasingly generating key insights for the development of precision approaches in heath and disease. Omics technologies have unlocked the high-throughput discovery of diagnostic biomarker and the systems-level evaluation of the efficacy and toxicity of novel therapies. In the subfield of sport medicine and sport science, the application of -omics methods is relatively recent; yet, the number of peer-reviewed original research articles has seen a steady rise over the last years. Athletic performance can be affected by a combination of endogenous and exogenous factors, including genomic and metabolomic profiles, as well as diet, medicament/drug intake, training regimen, and exercise. In this respect, the application of omics approaches encompassed, for instance, the multi-parametric assessment of training efficacy, injury predisposition, and the identification of robust biomarkers for the indirect detection of potentially “invisible” forms of doping. In addition, multi-omics integration approaches in sport medicine and doping sciences hold a clear promise for better understanding of the molecular/system levels of athlete’s pathophysiology. What we believe should be indispensable to maximize the potential benefit that such approaches promise, is the promotion of cross-disciplinary and collaborative science, beyond the individual laboratories and research institutions