Mass-spectrometry-based assays nowadays play an essential role in biomedical research and clinical applications. There are different types of commercial mass spectrometers on the market today, and triple quadrupole (QqQ) is one of the time-honored systems. Here, we overview the main areas of QqQ applications in biomedicine and assess the current level, evolution, and trends in the use of QqQ in these areas. Relevant data were extracted from the Scopus database using the specified terms and Boolean operators defined for each field of the QqQ application. We also discuss the recent advances in QqQ and QqQ-based analytical platforms, which promote the clinical application of these systems, and explain the indicated substantial increase in triple quadrupole use in biomedicine. The number of biomedical studies utilizing QqQ increased 2–3 times this decade. Triple quadrupole is most intensively used in the field of endocrine research and testing. On the contrary, the relative rate of immunoassay utilization—a major competitor of chromatography–mass spectrometry—decreased in this area as well as its use within Therapeutic drug monitoring (TDM) and forensic toxicology. Nowadays, the applications of high-resolution accurate mass (HRAM) mass spectrometers in the investigated areas represent only a small fraction of the total amount of research using mass spectrometry; however, their application substantially increased during the last decade in the untargeted search for new biomarkers.
According to the World Health Organization, ischemic stroke is the second leading cause of death in the world. Frequently, it is caused by brachiocephalic artery (BCA) atherosclerosis. Timely detection of atherosclerosis and its unstable course can allow for a timely response to potentially dangerous changes and reduce the risk of vascular complications. Omics technologies allow us to identify new biomarkers that we can use in diagnosing diseases. This research included 90 blood plasma samples. The study group comprised 52 patients with severe atherosclerotic lesions BCA, and the control group comprised 38 patients with no BCA atherosclerosis. Targeted and panoramic lipidomic profiling of their blood plasma was carried out. There was a statistically significant difference (p < 0.05) in the values of the indices saturated fatty acids (FAs), unsaturated FAs, monounsaturated FAs, omega-3, and omega-6. Based on the results on the blood plasma lipidome, we formed models that have a fairly good ability to determine atherosclerotic lesions of the brachiocephalic arteries, as well as a model for identifying unstable atherosclerotic plaques. According only to the panoramic lipidome data, divided into groups according to stable and unstable atherosclerotic plaques, a significant difference was taken into account: p value < 0.05 and abs (fold change) > 2. Unfortunately, we did not observe significant differences according to the established plasma panoramic lipidome criteria between patients with stable and unstable plaques. Omics technologies allow us to obtain data about any changes in the body. According to our data, statistically significant differences in lipidomic profiling were obtained when comparing groups with or without BCA atherosclerosis.
According to the World Health Organisation ischemic stroke is the second leading cause of death in the world. More frequent it is caused by brachiocephalic arteries (BCA) atherosclerosis. Timely detection of the atherosclerosis and it`s unstable course can allow a timely response to potentially dangerous changes and reduce the risk of vascular complications. Omics technologies allow us to identify new biomarkers that we can use in diagnosing diseases. The research included 90 blood plasma samples. The first group - 52 patients with severe atherosclerotic lesions, second group - 38 patients with no BCA atherosclerosis. Targeted and panoramic lipidomic profiling of blood plasma was carried out. There is a statistically significant difference (р
Background: Atherosclerosis and its associated chronic inflammation of the arterial wall disrupt fatty acid metabolism, leading to changes in plasma fatty acid composition. These alterations can be used to improve disease diagnosis and risk stratification by the development and application of specific lipidomic indices. Objectives: The objectives of this study are to evaluate the performance of conventional fatty acid indices and enhance diagnostic efficiency in atherosclerosis by introducing novel index based on plasma PUFA n-6 and n-3 content (Omega-6/3 Balance Index, O6/3-BI), as well as the perspective SFA/MUFA ratio (stearic/oleic acid ratio, C18:0/C18:1n-9) and a logit function combining PUFA and SFA/MUFA biomarkers. Methods: Plasma fatty acids were quantified by LC-MS/MS in healthy controls (n = 50) and patients with carotid atherosclerosis (n = 52), stratified by atorvastatin, rosuvastatin, or no statin therapy. The conventional indices (the Omega-3 Status (EPA + DHA), AA/EPA, and the omega-6/omega-3 ratio), and pathway ratios (C18:0/C18:1n-9; and C20:4n-6/C22:4n-6), as well as the newly introduced PUFA index and combined PUFA-SFA/MUFA logit function, were calculated. Their diagnostic performance for distinguishing atherosclerosis was assessed by a receiver operating characteristic (ROC) analysis with the cross-validation and calculation of Cliff’s Δ effect size. Results: The conventional parameters demonstrated a poor to low discrimination ability of the atherosclerosis patients’ groups from healthy controls (area under the ROC curve, AUC 0.548–0.711). In statin-treated patients, these conventional markers lost significance. The newly introduced PUFA index and SFA/MUFA ratio demonstrated improved patients’ discrimination with AUC 0.734–0.780 for the former and strong predictive power with AUC 0.831–0.858 for the latter marker and maintained their diagnostic value under statin therapy. The most significant positive effect size was observed for the SFA/MUFA ratio with Cliff’s Δ = 0.67–0.71. The combined PUFA-SFA/MUFA logit function also demonstrated a strong predictive power with AUC = 0.880 (Cliff’s Δ = −0.76), outperforming any single index. Conclusions: The newly introduced lipidomic index based on the PUFA content, SFA/MUFA ratio, and a logit function combining PUFA-SFA/MUFA biomarkers demonstrated a substantially better discrimination of atherosclerosis-related fatty acid metabolic disturbances than conventional fatty acid biomarkers.
Synthesis of steroid drugs is possible through biotechnological methods, however, the low efficiency of transporting steroid substrates across microbial membranes is the challenge in using microorganisms for their production. STARD1 and STARD3 proteins (members of the START domain family) work in some mammalian cells in conjunction with the steroidogenic P450scc enzyme system (cytochrome P450scc, adrenodoxin reductase, and adrenodoxin), transporting cholesterol to the mitochondria, where cytochrome P450scc converts it into pregnenolone. In this study, we investigated the effect of STARD1 and STARD3 on P450scc efficiency in E. coli expression models. The combination of the STARD3(216–444) or STARD1(53–285) protein bearing an N-terminal periplasm-targeting sequence, with the P450scc system reconstituted in E. coli, resulted in an increase of pregnenolone synthesis by 3.2- and 1.9-fold, respectively, on a laboratory scale. STARD3(216–444) showed higher levels of expression and a greater effect on the incorporation of sterols into cells and whole-cell biotransformation of cholesterol, compared to STARD1(53–285). This study proposes a fundamentally new approach to optimizing the functioning of the P450scc system in microbial cells, which uses the cholesterol transfer protein to increase the uptake efficiency of a poorly soluble steroid substrate by bacteria. The demand for steroid medications is increasing, and the use of specific carrier proteins could be a useful tool to enhance the efficiency of whole-cell biosynthesis of various steroid compounds.