Clinica Chimica Acta最新文献

{"title":"Enhancing the detection sensitivity of mNGS in Bronchoalveolar Lavage Fluid through cell counting: An empirical study","authors":"Zhe Liu ,&nbsp;Shangdong Yang ,&nbsp;Shumei Xie ,&nbsp;Depan Cao ,&nbsp;Wen Xi ,&nbsp;Yang Xiao ,&nbsp;Xin Xu ,&nbsp;Zhonglin Wang ,&nbsp;Lifeng Li ,&nbsp;Jian Hu ,&nbsp;Xiaoqin Wang","doi":"10.1016/j.cca.2025.120311","DOIUrl":"10.1016/j.cca.2025.120311","url":null,"abstract":"<div><h3>Objectives</h3><div>Lower respiratory tract infections pose significant clinical challenges due to their high morbidity and mortality rates. While metagenomic next-generation sequencing (mNGS) has emerged as a promising diagnostic tool, its sensitivity is often compromised by host DNA contamination that overwhelms microbial signals. Selective host DNA depletion through cell lysis effectively reduces host DNA; however, it has an impact on microorganisms with relatively thin cell walls, and samples with low host content may introduce more environment or reagent-derived microbial contamination, interfering the detection results. Methods for determining host DNA depletion based on sample type, sample characteristics or using spike-in controls to monitor sensitivity do not fully consider the potential limitations of host depletion technology on microbial detection, nor do they evaluate the possible significant impact on detection efficiency. This study aimed to develop a pre-analytical method for accurate host DNA content assessment.</div></div><div><h3>Methods</h3><div>We established a cell-counting-based method for precise cellular content measurement in clinical bronchoalveolar lavage fluid (BALF) samples. The protocol involved: (1) evaluating the linearity and robustness of cell-counting dyes in BALF samples with varying characteristics, (2) assessing the correlation between cell counts and extracted nucleic acid mass, (3) investigating cellular counting thresholds for host depletion in clinical BALF analysis, and (4) implementing the optimized cell-counting method in clinical mNGS testing to guide selective-lysis treatment.</div></div><div><h3>Results</h3><div>Acridine orange/propidium iodide (AO/PI) staining demonstrated superior performance compared to trypan blue and 4',6-diamidino-2-phenylindole (DAPI), particularly in turbid and bloody BALF samples. Implementing a host depletion threshold at 1 × 10⁶ cell counts significantly improves pathogen detection rates in high host background samples, while effectively preserving the detection sensitivity for pathogens in moderate and low host background samples.</div></div><div><h3>Conclusions</h3><div>Our findings demonstrate that cell counting serves as a reliable pre-analytical tool for determining optimal selective-lysis treatment in BALF mNGS testing, enhancing diagnostic accuracy while preserving pathogen integrity.</div></div>","PeriodicalId":10205,"journal":{"name":"Clinica Chimica Acta","volume":"574 ","pages":"Article 120311"},"PeriodicalIF":3.2,"publicationDate":"2025-04-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143903909","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Advances in Z-drug detection","authors":"Selen Al,&nbsp;Aykut Kul,&nbsp;Olcay Sagirli","doi":"10.1016/j.cca.2025.120329","DOIUrl":"10.1016/j.cca.2025.120329","url":null,"abstract":"<div><div>The use of Z-drugs (zolpidem, zaleplon, zopiclone, and eszopiclone) has increased in recent years, leading to a growing need for analytical methods to accurately and reliably detect these compounds in biological samples. This review comprehensively presents the latest developments in the analysis of Z-drugs in biological samples, examining methods such as LC-MS/MS, GC–MS/MS, and HPLC. These methods have been thoroughly evaluated in biological matrices (such as blood, plasma, serum, urine, and hair), with particular focus on characteristics like sample preparation technique, calibration range, LLOQ, analysis time, mobile phase, and column. Recent studies have shown that these methods are effective in routine toxicological analysis and forensic detection of drug abuse. This review aims to provide a valuable resource for researchers and practitioners involved in the detection and analysis of Z-drugs, contributing to a better understanding and management of both therapeutic and illicit use of these compounds.</div></div>","PeriodicalId":10205,"journal":{"name":"Clinica Chimica Acta","volume":"574 ","pages":"Article 120329"},"PeriodicalIF":3.2,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143878450","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Innovative electrochemical biosensors for tuberculosis detection","authors":"Mohammad Hassan Sadeghi ,&nbsp;Safa Radmehr ,&nbsp;Neda Mohagheghzadeh ,&nbsp;Javad Fathi ,&nbsp;Yalda Malekzadegan ,&nbsp;Hesam Zendehdel Moghadam","doi":"10.1016/j.cca.2025.120327","DOIUrl":"10.1016/j.cca.2025.120327","url":null,"abstract":"<div><div>Tuberculosis (TB) continues to pose a significant global health threat, highlighting the urgent need for the development of rapid, precise, and accessible diagnostic tools to effectively manage its transmission. Conventional diagnostic techniques, such as sputum microscopy and culture-based assays, face several drawbacks, including lengthy processing times, limited sensitivity, and the requirement for specialized laboratory facilities. In this landscape, electrochemical biosensors have emerged as promising alternatives, offering improved sensitivity, specificity, and rapid detection capabilities. This review presents a thorough overview of recent advancements in the development and application of innovative electrochemical biosensors for TB detection. It explores the integration of nanomaterials such as graphene, gold nanoparticles, and carbon nanotubes, focusing on their contributions to enhanced sensor performance in terms of signal amplification and biorecognition efficacy. By synthesizing current research and technological developments, this review emphasizes the considerable potential of electrochemical biosensors to transform TB diagnostics, ultimately assisting in better disease management and control strategies worldwide.</div></div>","PeriodicalId":10205,"journal":{"name":"Clinica Chimica Acta","volume":"574 ","pages":"Article 120327"},"PeriodicalIF":3.2,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143873424","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Relative change rate of anti-acetylcholine receptor antibodies concentration Correlates with the recurrence of myasthenia gravis in AChR-Abs-positive patients","authors":"Jingluan Tian ,&nbsp;Xiaoling Zhou ,&nbsp;Feng Zhu ,&nbsp;Xinyi Xiao ,&nbsp;Hairong Ma ,&nbsp;Xin Wang ,&nbsp;Yanzheng Gu ,&nbsp;Yueping Shen ,&nbsp;Xingshun Xu ,&nbsp;Qun Xue","doi":"10.1016/j.cca.2025.120330","DOIUrl":"10.1016/j.cca.2025.120330","url":null,"abstract":"<div><h3>Backgrounds</h3><div>Myasthenia gravis (MG) is an autoimmune disease predominantly caused by anti-acetylcholine receptor antibodies (AChR-Abs). In this study, the relationship between MG relapse and AChR-Abs concentration were investigated.</div></div><div><h3>Methods</h3><div>We retrospectively reviewed the clinical records of 41 patients with AChR-Abs-positive MG. Data collected included age, sex, MG status within 12 months (untreated status, UTMG; recurrence status, RCMG), MG subgroups, QMGs, AChR-Abs concentration and treatments. The relative change rate (RCR) of AChR-Abs concentration was calculated as: RCR(%)=(AChR-Abs–AChR-Abs_UTMG)/AChR-Abs_UTMG/month*100. ROC curve analysis and binary logistic regression analyses were used to evaluate the association between these factors and MG recurrence.</div></div><div><h3>Results</h3><div>Within 12 months, 12 patients experienced a recurrence (RCMG). A weak correlation was observed between AChR-Abs level in UTMG and QMGs (r = 0.336, <em>p</em> = 0.032). MG subgroups, QMGs, AChR-Ab levels and RCR differed significantly between RCMG and non-RCMG group. Single factor ROC curves indicated that AChR-Abs concentration, RCR and MG subgroups were related to MG recurrence (AUC = 0.724, 0.736, 0.736). Combining with RCR and MG subgroups, the ROC yielded an AUC of 0.899, demonstrating high sensitivity and specificity.</div></div><div><h3>Conclusion</h3><div>RCR of AChR-Abs was correlated with MG progression, and repeated AChR-Abs measurements can assist in monitoring disease status and guiding therapeutic decision-making.</div></div>","PeriodicalId":10205,"journal":{"name":"Clinica Chimica Acta","volume":"574 ","pages":"Article 120330"},"PeriodicalIF":3.2,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143899290","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Electrochemical biosensors for hepatocellular carcinoma","authors":"Lei Wang ,&nbsp;Jianjiang Pan ,&nbsp;Bita Badehnoosh","doi":"10.1016/j.cca.2025.120328","DOIUrl":"10.1016/j.cca.2025.120328","url":null,"abstract":"<div><div>The current review analyzes progress in electrochemical detection techniques for hepatocellular carcinoma biosignatures, highlighting their potential to enhance the timely detection and management of hepatocellular carcinoma. In this study, the authors explore the present state of hepatocellular carcinoma biosignatures, encompassing conventional proteins such as alpha-fetoprotein and promising biosignatures like non-coding RNAs and circulatory tumor DNA (ctDNA). This text analyzes the principles of electrochemical biosensing and explores sophisticated sensor designs employing surface modification techniques, innovative recognition elements, and nanomaterials. Particular focus is directed towards aptamer-based sensors, microfluidic technologies, and label-free methodologies. Herein, recent advancements in enhancing sensitivity and specificity are discussed, with some platforms reaching a threshold at the femtogram scale. The discussion also encompasses the progress achieved in point-of-care applications and the obstacles faced in transitioning experimental paradigms to medical applications. The prospective influence of these methodologies on medical results is under evaluation, emphasizing early detection and tailored treatment approaches. Future research should focus on creating advanced, integrated detection systems and conducting comprehensive clinical validation studies to assess the real-world effectiveness of electrochemical biosensors.</div></div>","PeriodicalId":10205,"journal":{"name":"Clinica Chimica Acta","volume":"574 ","pages":"Article 120328"},"PeriodicalIF":3.2,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143896094","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A machine learning approach for assessing acute infection by erythrocyte sedimentation rate (ESR) kinetics","authors":"Andrea Padoan ,&nbsp;Ilaria Talli ,&nbsp;Michela Pelloso ,&nbsp;Luisa Galla ,&nbsp;Francesca Tosato ,&nbsp;Daniela Diamanti ,&nbsp;Chiara Cosma ,&nbsp;Elisa Pangrazzi ,&nbsp;Alessandra Brogi ,&nbsp;Martina Zaninotto ,&nbsp;Mario Plebani","doi":"10.1016/j.cca.2025.120308","DOIUrl":"10.1016/j.cca.2025.120308","url":null,"abstract":"<div><h3>Background</h3><div>The erythrocyte sedimentation rate (ESR) is a traditional marker of inflammation, valued for its simplicity and low cost but limited by unsatisfactory specificity and sensitivity. This study evaluated the equivalence of ESR measurements obtained from three automated analyzers compared to the Westergren method. Furthermore, various machine learning (ML) techniques were employed to assess the usefulness of early sedimentation kinetics in inflammatory disease classification.</div></div><div><h3>Methods</h3><div>A total of 346 blood samples from control, rheumatological, oncological, and sepsis/acute inflammatory status groups were analyzed. ESR was measured using TEST 1 (Alifax Spa, Padua, Italy), VESMATIC 5 (Diesse Diagnostica Senese Spa, Siena, Italy), CUBE 30 TOUCH (Diesse Diagnostica Senese Spa, Siena, Italy) analyzers, and the Westergren method. Early sedimentation rate kinetics (within 20 min) obtained with the CUBE 30 TOUCH were assessed. ML models [Gradient Boosting Machine (GBM), Support Vector Machine (SVM), Naïve Bayes (NB), Neural Networks (NN) and logistic regression (LR)] in discriminating groups were trained and validated using ESR, sedimentation slopes, and clinical data. A second validation cohort of control and sepsis samples was used to validate LR models.</div></div><div><h3>Results</h3><div>Automated methods showed good agreement with Westergren’s results. Multivariate analyses identified significant associations between ESR values (measured by CUBE 30 TOUCH) and age (p = 0.025), gender (p &lt; 0.001), and, overall, with samples’ group (p &lt; 0.001). Sedimentation rate slopes differed significantly across groups, particularly between 12 and 20 min, with sepsis cases showing distinct patterns. ML models achieved moderate accuracy, with GBM performing best (AUC 0.800). LR for sepsis classification in the validation cohort achieved an AUC of 0.884, with high sensitivity (96.9 %) and specificity (74.2 %). In the second validation cohort, LR outperformed prior results, reaching an AUC of 0.991 (95 % CI: 0.973–1.000), with 95.2 % sensitivity and 100 %.</div></div><div><h3>Conclusions</h3><div>Current automated technologies for ESR measurement well agree with the reference method and provide robust results for evaluating systemic infections. The novelty of this study lies in connecting ESR sedimentation kinetics to disease states, particularly for identifying sepsis/acute inflammatory status. Future studies with larger datasets are needed to validate these approaches and guide clinical application.</div></div>","PeriodicalId":10205,"journal":{"name":"Clinica Chimica Acta","volume":"574 ","pages":"Article 120308"},"PeriodicalIF":3.2,"publicationDate":"2025-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143865231","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Macrocomplexes: An establishing critical limits of post-PEG serum enzymes and proteins in adults and children","authors":"Jitka Čásenská,&nbsp;Janka Franeková,&nbsp;Antonín Jabor","doi":"10.1016/j.cca.2025.120326","DOIUrl":"10.1016/j.cca.2025.120326","url":null,"abstract":"<div><h3>Background</h3><div>Exclusion of serum macroform presence is essential to avoid potential misdiagnosis. The aim of this study was to determine critical limits (CL) for enzymes (total and pancreatic amylase, AMY and pAMY; aminotransferases, AST and ALT; alkaline phosphatase, ALP; creatine kinase, CK; lipase, LIP; gamma-glutamyl transferase, GGT; lactate dehydrogenase, LDH) after polyethylene glycol (PEG) precipitation in children and adults.</div></div><div><h3>Methods</h3><div>A total of 126 sera of patients suspected for macroenzyme presence (adults or children) was matched by serum enzyme activity of controls and all these sera were precipitated by PEG. PEG-precipitable activity ( %PPA) and CLs of %PPA were calculated for adults, children, and controls.</div></div><div><h3>Results</h3><div>CLs of %PPA were substantially lower than common 60 % for ALP and CK in all subjects. Significant differences between adults and children were found for ALT and ALP (lower in children), and LIP and CK (higher in children). No meaningful correlation was found between initial enzyme activity and %PPA.</div></div><div><h3>Conclusions</h3><div>Age specific CLs should be used for ALP, ALT, CK, and LIP. Generally accepted value of %PPA of 60 % may be used with caution for AMY, pAMY, AST, GGT, and LDH. A careful selection of CLs is needed for timely detection of macrocomplexes.</div></div>","PeriodicalId":10205,"journal":{"name":"Clinica Chimica Acta","volume":"574 ","pages":"Article 120326"},"PeriodicalIF":3.2,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143870416","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Determination of human urinary cortisol, cortisone and relative phase II metabolites by dilute-and-shoot LC-MS/MS analysis: an application to adrenal disease","authors":"Laura Leoni ,&nbsp;Mirko Parasiliti Caprino ,&nbsp;Giulia Montesano ,&nbsp;Chiara Lopez ,&nbsp;Martina Bollati ,&nbsp;Fabio Settanni ,&nbsp;Ezio Ghigo ,&nbsp;Giulio Mengozzi ,&nbsp;Federico Ponzetto","doi":"10.1016/j.cca.2025.120324","DOIUrl":"10.1016/j.cca.2025.120324","url":null,"abstract":"<div><h3>Background</h3><div>Cortisol and its primary metabolite, cortisone, play key roles in managing stress responses and maintaining balance via the hypothalamic–pituitary–adrenal axis. Traditional immunoassays, often hindered by cross-reactivity, can yield inaccurate results. This study introduces a novel dilute-and-shoot using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) for the rapid and accurate analysis of urinary free cortisol, cortisone, and their intact phase II metabolites.</div></div><div><h3>Methods</h3><div>A new LC-MS/MS method was developed to quantify five analytes, including intact glucuro- and sulfo-conjugated cortisol and cortisone metabolites. The method underwent validation assessments in accordance with ISO 17025 standards for quantitative methods. Its performance was compared with a commercial LC-MS/MS assay used in clinical laboratories, analyzing free cortisol and cortisone in 79 24-hour urine samples. Additionally, the novel method was applied to authentic urine samples from 168 adrenal disease patients.</div></div><div><h3>Results</h3><div>The validation protocol assessed method’s selectivity, matrix effects, quantitative performance and carry-over. The comparison showed excellent correlation with the commercial LC-MS/MS method for urinary free cortisol and cortisone measurement. Pathological sample analysis indicated that monitoring intact phase II metabolites, particularly sulfoconjugates, could aid in differentiating adrenal diseases.</div></div><div><h3>Conclusion</h3><div>The developed and validated dilute-and-shoot LC-MS/MS method demonstrated satisfactory performance, offering a faster, cost-effective solution for adrenal hormone analysis in clinical settings. This research also provided insights into adrenal function by examining the urinary excretion of target analytes in a pathological cohort, enhancing diagnostic precision for related disorders.</div></div>","PeriodicalId":10205,"journal":{"name":"Clinica Chimica Acta","volume":"574 ","pages":"Article 120324"},"PeriodicalIF":3.2,"publicationDate":"2025-04-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143870415","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Assessment of the performance of a dual-sample oral cholate challenge test: The HepQuant DuO test","authors":"Michael P. McRae ,&nbsp;Touraj Shokati ,&nbsp;Uwe Christians ,&nbsp;Steve M. Helmke ,&nbsp;Gregory T. Everson","doi":"10.1016/j.cca.2025.120325","DOIUrl":"10.1016/j.cca.2025.120325","url":null,"abstract":"<div><h3>Background</h3><div>Liver health is currently evaluated using nonspecific blood tests, fibrosis surrogates, and invasive procedures, none of which directly measure liver function and physiology. The validated HepQuant SHUNT test quantifies liver function and physiology and is linked to clinical outcomes. Herein we present the reliability of a simplified version, the dual-sample oral cholate challenge (HepQuant DuO) test.</div></div><div><h3>Methods</h3><div>Samples from the SHUNT-V study representing various liver disease etiologies were used in experiments to assess test reliability. We compared the HepQuant DuO test analyte measurements relative to the previously validated HepQuant SHUNT test. Precision and imprecision (%CV) of analyte measurements and HepQuant DuO test parameters were evaluated over four days, with two runs per day and three replicates per run, using two-way nested hierarchical ANOVA. Analytical specificity was evaluated in samples spiked with endogenous ¹²C-cholic acid.</div></div><div><h3>Results</h3><div>All samples met the prespecified method comparison acceptance criteria with a mean bias of 2.6 %, demonstrating equivalency between the analytical methods of HepQuant DuO and SHUNT. Total imprecision across all HepQuant DuO test parameters, for both 13C- and d4-cholate, ranged from 0.6 % to 2.1 %. Samples spiked with high concentrations of ¹²C-cholic acid demonstrated lack of interference with a percent change relative to controls of 3.2 % or less.</div></div><div><h3>Conclusions</h3><div>The studies described herein established the reliability of the HepQuant DuO test in terms of precision, analytical specificity, and agreement with the validated HepQuant SHUNT test. Reportable ranges and reference intervals are also described.</div></div>","PeriodicalId":10205,"journal":{"name":"Clinica Chimica Acta","volume":"574 ","pages":"Article 120325"},"PeriodicalIF":3.2,"publicationDate":"2025-04-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143865230","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabolic origin and significance of 3-methylglutaryl CoA","authors":"Elizabeth A. Jennings,&nbsp;Zane H. Abi-Rached,&nbsp;Robert O. Ryan","doi":"10.1016/j.cca.2025.120320","DOIUrl":"10.1016/j.cca.2025.120320","url":null,"abstract":"<div><div>3-Methylglutaryl (3MG) CoA is not part of any biochemical pathway, yet its byproducts, 3MG carnitine and 3MG acid, are disease biomarkers. Both compounds are excreted in HMG CoA lyase deficiency, while 3MG aciduria occurs in inborn errors of metabolism (IEM) associated with compromised mitochondrial energy metabolism. In one such disorder (i.e., TMEM70 deficiency), 3MG carnitine is also present. Moreover, in a number of chronic and acute maladies, elevated levels of 3MG carnitine are present. The precursor of 3MG CoA is<!--> <em>trans</em>-3-methylglutaconyl (3MGC) CoA. When<!--> <em>trans</em>-3MGC CoA levels rise, a portion of this metabolite pool is reduced to 3MG CoA, potentially via a side reaction involving glutaryl CoA dehydrogenase (GCDH), which normally catalyzes the oxidative decarboxylation of glutaryl CoA to crotonyl CoA and CO₂. This reaction occurs via a two-step process wherein glutaryl CoA is initially oxidized to glutaconyl CoA, coupled to reduction of the enzyme’s FAD prosthetic group. Enzyme-bound glutaconyl CoA is then decarboxylated to the reaction product, crotonyl CoA. Before GCDH can accept another glutaryl CoA the flavin prosthetic group must be oxidized to FAD by donating electrons to electron transferring flavoprotein (ETF). However, genetic- or disease-induced defects in electron transport chain function can impede this reaction. We propose that<!--> <em>trans</em>-3MGC CoA is a substrate for reduced GCDH and, when glutaryl CoA and<!--> <em>trans</em>-3MGC CoA are present, GCDH is able to bypass ETF and cycle between oxidized and reduced states, producing crotonyl CoA and CO₂ <!-->from glutaryl CoA, and 3MG CoA from<!--> <em>trans</em>-3MGC CoA.</div></div>","PeriodicalId":10205,"journal":{"name":"Clinica Chimica Acta","volume":"574 ","pages":"Article 120320"},"PeriodicalIF":3.2,"publicationDate":"2025-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143848051","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}