Serum and urine protein electrophoresis have long served as the mainstay for monitoring monoclonal protein (M-protein) levels in assessing treatment response for multiple myeloma (MM). Despite limitations in sensitivity and specificity, these methods remain the primary clinical detection techniques.

Melting curve analysis following qPCR allows for the efficient and cost-effective detection of multiple targets. By examining the shape and number of peaks in the melting curve, multiple targets can be simultaneously detected within a single tube. The integration of melting curve analysis with various probe technologies continues to advance the capabilities of molecular testing in pathogen detection. This paper reviews the principles and development of qPCR-melting curve analysis in molecular diagnostics and explores future trends in this analytical technique.

The accuracy of urine albumin results is important for healthcare decisions related to kidney disease. A commutable human urine secondary certified reference material (CRM) is needed for metrological traceability of results from clinical laboratory in vitro diagnostics (IVD) measurement procedures (MPs). The commutability of NIST Standard Reference Material® (SRM) 3666, Albumin and Creatinine in Frozen Human Urine, was assessed using the NIST reference measurement procedures (RMPs) and IVD-MPs for urine albumin and creatinine. Fifty single-donor clinical samples and NIST SRM 3666 (Level I to Level IV) were measured using IVD-MPs from Abbott Laboratories, Beckman Coulter, Mindray, QuidelOrtho, Roche Diagnostics, and Siemens Healthcare Diagnostics. Commutability was assessed using the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) approach based on differences in bias between the SRM and clinical samples. For urine creatinine (Enzymatic and Jaffé), all levels of SRM 3666 were commutable with clinical samples on all IVD-MPs, with the exception of 1 IVD-MP for urine creatinine-Jaffé with Level III. For urine albumin, Level I is commutable with clinical samples for use with 5 IVD-MPs; Level II is commutable for use with 6 IVD-MPs; Level III is commutable for use with 6 IVD-MPs; and Level IV is commutable for use with 3 of the 5 IVD-MPs that could measure Level IV. This study demonstrated that NIST SRM 3666 has commutability properties suitable for use on select IVD-MPs for urine albumin, as well as for all examined Enzymatic and Jaffé IVD-MPs for urine creatinine.

Ovarian cancer remains one of the most lethal gynecologic malignancies, largely because of late-stage diagnosis and the absence of reliable biomarkers for early detection and therapeutic stratification. Recent advances in high-throughput technologies have enabled multi-omics approaches that integrate genomics, transcriptomics, proteomics, metabolomics, and epigenomics to elucidate the comprehensive molecular landscape of ovarian cancer. This narrative review synthesizes current progress in applying multi-omics strategies to biomarker discovery, highlighting how integrative analyses uncover novel diagnostic, prognostic, and predictive candidates beyond the limitations of single-omics studies. We discuss methodological frameworks, computational pipelines and translational challenges in harmonizing heterogeneous datasets, as well as the potential of systems biology and machine learning to improve biomarker validation. Particular emphasis is placed on the identification of noncoding RNAs, protein signatures, and metabolic alterations as promising biomarker classes. Finally, we outline future directions for clinical implementation, including the development of multiparameter biomarker panels and precision medicine applications. By bridging molecular complexity with translational utility, multi-omics approaches hold transformative potential to advance biomarker identification and improve patient outcomes in ovarian cancer.

Distinguishing bacterial from viral respiratory infections is challenging in Emergency Department (ED), driving diagnostic uncertainty and antibiotic overuse. The MeMed BV test measures three host protein markers and combines them into a single numeric score. This study evaluates its analytical and clinical performance and assesses its potential to improve antibiotic stewardship.

Urinary tract infections (UTIs) are among the most prevalent bacterial infections worldwide, often presenting diagnostic challenges owing to nonspecific symptoms and limitations of conventional biomarkers. Recent advances in EV research have highlighted urinary exosome nanosized vesicles secreted by renal and urothelial cells as promising noninvasive biomarkers for UTI detection and monitoring. These exosomes encapsulate a rich cargo of proteins, lipids, and nucleic acids reflective of the physiological and pathological state of the urinary tract. This review synthesizes current evidence on the diagnostic potential of urinary exosomes in UTIs, emphasizing their role in pathogen recognition, immune modulation, and inflammation signaling. We discuss emerging methodologies for exosome isolation and characterization and evaluate their clinical applicability in differentiating bacterial strains, predicting disease severity, and guiding antimicrobial therapy. By integrating molecular insights with translational perspectives, urinary exosomes may redefine the diagnostic landscape of UTIs and pave the way for precision urological care.

Long non-coding RNAs (lncRNAs) have emerged as useful diagnostic and prognostic markers as well as therapeutic targets in cancer. Dysregulated levels of lncRNAs have been described in various cancers including breast cancer (BC). Our study aims to screen novel lncRNAs involved in BC and investigate any possible role in helping diagnosis and determining prognosis of BC patients.

External Quality Assessment (EQA) is essential for ensuring inter-laboratory comparability, yet the interpretability of EQA data depends critically on the commutability of control materials (CM). This study assessed the commutability of CM used in the Lombardy Regional EQA program for selected clinical chemistry measurands.

The emergence of veterinary sedatives such as xylazine and medetomidine as adulterants in the illicit drug supply poses increasing challenges to clinical toxicology and public health. Medetomidine, a potent alpha-2 adrenergic receptor agonist not approved for human use, has recently been detected in overdose cases, particularly in fentanyl-positive individuals. To address this, we developed and validated a liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay capable of detecting xylazine and medetomidine in urine with minimal sample preparation, a 60-min hydrolysis incubation, and a 7-min chromatographic run time. Analytical validation demonstrated acceptable precision, accuracy, linearity, and minimal matrix effects. We then analyzed 100 retrospective urine samples from a polydrug-using population. Xylazine was detected in 20 % and 3-hydroxymedetomidine (medetomidine's primary metabolite in urine) in 12 % of samples, with both compounds most frequently co-occurring with fentanyl. A subset of medetomidine-positive cases also tested positive for xylazine, indicating potential co-adulteration. This study highlights the value of expanded toxicological testing in identifying emerging adulterants that may otherwise go undetected by routine screening.

Trisomy 21 (T21) is the genetic cause of Down syndrome (DS), and the presence of extra genetic material causes altered expression of genes located on chromosome 21 (Hsa21) and others, with effects as altered levels of metabolic reaction products. The one‑carbon pathway plays a central role in correct human neurodevelopment and was found to be altered in DS and neurological impairments of different entities. In this work, the expression of 42 genes involved in the one‑carbon cycle was analyzed in blood samples from 10 subjects with T21 and 10 euploid (N) subjects. Additionally, plasmatic concentration of methylcobalamin (MeCbl) was evaluated in 10 subjects with T21 and 7 N subjects. The results showed that 13 genes out of 42 were differentially expressed: 11 were over-expressed (ABCC3, ABCC4, ARMT1, CTH, FOLR2, GART, ICMT, PRMT2, SETD4, SLC19A1, and NSD2) and 2 were under-expressed (NSUN3 and TRMT112). Among these, 4 over-expressed genes are located on Hsa21 (GART, PRMT2, SETD4, and SLC19A1). Statistical analyses revealed significant correlations between gene expression data, highlighting interconnections among genes. Finally, MeCbl shows a slight statistically significant reduction in the T21 group. In conclusion, the presence of three copies of Hsa21 leads to the dysregulation of gene expression associated with the one‑carbon cycle. This dysregulation affects genes located on both Hsa21 and other chromosomes resulting in metabolic alterations. Additionally, new gene interconnections were discovered that have not been previously reported in the literature.

Cardiovascular disease (CVD) remains the leading cause of mortality globally, necessitating the development of rapid, accurate, and accessible diagnostic tools for early detection and effective management. Electrochemical aptamer-based biosensors have emerged as promising candidates in this domain because of their high sensitivity, specificity, and potential for miniaturization and real-time monitoring. This narrative review provides a comprehensive overview of the current state of electrochemical aptamer-based biosensors applied to cardiovascular disease diagnostics. This review highlights recent advancements in sensor design, such as nanomaterial integration and signal amplification strategies, which have significantly enhanced the performance of these biosensors. Additionally, we examine the application of these biosensors in detecting key cardiovascular biomarkers, such as troponins, C-reactive protein, and myoglobin, underscoring their clinical relevance. By synthesizing current knowledge and technological innovations, this review underscores the potential of electrochemical aptamer-based biosensors to revolutionize cardiovascular disease diagnostics and improve patient outcomes.

Preeclampsia (PE) is a major contributor to maternal morbidity and mortality, particularly in low- and middle-income countries. Women with a history of PE face a higher long-term risk of developing cardiovascular disease, but the mechanisms underlying this association remain poorly understood. Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a key regulator of plasma low-density lipoprotein cholesterol and has been linked to atherosclerosis and metabolic dysfunction. This study aimed to evaluate plasma PCSK9 levels in women with a history of PE, compared to those with history of normotensive pregnancies, and to explore the relationship between PCSK9, LDLc, and markers of cardiovascular disease.

Chronic hepatitis B (CHB) currently lacks a rapid and objective inflammatory marker. However, matrix metalloproteinase-9 (MMP-9) is a biomarker for necroinflammation and fibrosis, as it is mechanistically linked to these processes. This review summarizes the data supporting the association between circulating MMP-9 and hepatic necroinflammation and assesses the use of electrochemical biosensors for quantification at the point of care (POC). MMP-9 is found in several molecular forms, including pro-MMP-9 (∼92 kDa) and active (∼82-83 kDa), with different clinical implications. Active MMP-9 represents the ongoing matrix remodelling whereas total MMP-9 represents the summed expression (r = 0.81 zymography vs enzyme-linked immunosorbent assay (ELISA)). However, a direct comparison of these forms in terms of histology and outcomes has not been performed. Treatment dynamics provide a good clinical background: serum MMP-9 drops 40-60 % in 6 months of antiviral therapy with normalization of MMP-9 4-8 weeks before alanine aminotransferase (ALT) making MMP-9 an early treatment response marker (≥50 % reduction predicts good outcome whereas ≤30 % predicts failure). Conformity to clinical needs requires Total Allowable Error (TEa) analysis with clinical decision limits for significant inflammation (≥G2) being ∼150-180, advanced fibrosis (F2) ∼200-220, and cirrhosis (F4) ∼300 ng/mL. Multi-platform standardization includes commuted secondary reference materials for CHB with stable pro/active ratios and external quality assessment programs that confirm multi-laboratory interoperability. As such, it is suggested that a four-step framework of validation that combines clinical thresholds, commutable reference materials, and EQA-harmonized reporting be employed to ensure reliable MMP-9 biosensing for monitoring hepatitis B worldwide.

Maple syrup urine disease (MSUD), also called leucinosis, is a rare inborn error of metabolism characterized by the deficiency of an enzyme complex: branched-chain alpha-keto acid dehydrogenase (BCKAD). This complex is located in the mitochondria and is responsible for metabolizing the three essential branched-chain amino acids (BCAA): isoleucine, leucine, and valine. As the patient with MSUD cannot directly break down these "building blocks" of protein, a harmful build-up of substances in the plasma, urine and tissues will follow. In this review, we overview available data of inflammation in MSUD patients, cells and animals and discuss possible pathophysiological links. Available studies demonstrate that exposure to high levels of BCAAs leads to altered expression of inflammatory markers in cells, animals, and patients with MSUD. Proinflammatory cytokines such as IL-1β, IL-6, TNF-α, and IFN-γ are commonly elevated in specific brain regions, while anti-inflammatory responses (e.g., IL-10) may be suppressed. Additionally, increased expression of cell adhesion molecules (sICAM-1, sVCAM-1) and receptor for advanced glycation end-products (RAGE) has been observed. These findings suggest that inflammation may play an important role in MSUD pathophysiology. Further studies are needed to clarify the relationship between inflammatory imbalance, neurodegeneration, and clinical outcomes in MSUD.

Tree nut and peanut allergy is increasing in prevalence across Europe, yet the molecular sensitization patterns underlying these allergies remain incompletely defined in Southern Europe, particularly in Italy.

Physical exercise has emerged as a powerful modulator of molecular pathways involved in cancer development, progression, and therapeutic response. Recent evidence highlights the pivotal role of circulating and exosomal non-coding RNAs-including microRNAs (miRNAs) and long non-coding RNAs (lncRNAs)-as mediators linking exercise to systemic and tumor-specific adaptations. These molecules regulate key processes such as cell proliferation, apoptosis, angiogenesis, inflammation, and immune modulation, thereby influencing the tumor microenvironment and overall cancer trajectory. Exercise-induced alterations in circulating and exosomal miRNAs and lncRNAs have been shown to reflect molecular reprogramming at both systemic and cellular levels, suggesting their potential as minimally invasive biomarkers for cancer diagnosis, prognosis, prevention, and treatment monitoring. Furthermore, exosomal non-coding RNAs serve as intercellular communicators, transferring exercise-mediated molecular signals to target tissues, which may contribute to improved therapeutic efficacy and reduced tumor progression. This review synthesizes current evidence on how different exercise modalities modulate circulating and exosomal miRNAs and lncRNAs in cancer, emphasizing their diagnostic and prognostic potential and the underlying molecular mechanisms. Understanding these interactions provides novel insights into how physical activity can be harnessed as an adjuvant strategy in precision oncology, paving the way for predictive, preventive, and personalized approaches to cancer management.

The intensive exploitation of planet resources, driven by the rapid population growth and overconsumption, is threatening environmental and human sustainability, and a change of behaviour is urgently needed. The United Nations have defined 17 sustainable development goals that reflect this necessity and which intend to alert humanity about the importance of intervening in the diverse domains of human activity, including healthcare. Sensors and biosensors show advantageous attributes and great potential towards more sustainable healthcare practices, although some requirements must be assured, including the sustainability of all elements composing the device. In the case of affinity biosensors, the sustainability of the bioligand has a significant impact in the sustainability of the whole device. Lectins are used as bioligands in affinity biosensors for clinical applications that encompass the detection of particular glycostructures. This review aims to carefully analyse lectins' characteristics in order to deduce their sustainability profile for the development of sustainable devices for clinical sensing. A comparison with antibodies, as another type of natural bioligands frequently used for the same analytical applications is also presented.

Endometriosis diagnosis currently relies on invasive laparoscopy, creating a need for non-invasive alternatives. This study evaluates microRNAs (miRNAs) as potential diagnostic biomarkers for endometriosis through systematic evidence synthesis and performance analysis.

Triple-negative breast cancer is an aggressive subtype lacking hormone receptors and HER2, which limits therapeutic options and complicates early diagnosis. This pilot study aimed to evaluate the potential of electronic noses as a non-invasive tool for detecting TNBC by analyzing volatile organic compounds emitted by cell cultures. A total of 32 samples (22 TNBC MDA-MB-231 and 10 non-tumor controls) were analyzed using a MOX sensor-based electronic nose. Chemometric analyses including PCA, PLS-DA, and Random Forest were applied, with leave-one-out cross-validation and permutation testing to assess model robustness. PCA showed that the first two components explained 92.2 % of the variance and clearly separated TNBC from healthy samples. PLS-DA achieved 95 % classification accuracy (R = 0.81, Q = 0.76), and Random Forest validation confirmed high predictive performance. These findings highlight the ability of electronic noses to discriminate between tumor and non-tumor volatile profiles, supporting their use as a complementary tool for the early diagnosis of aggressive breast cancers.

Trichomoniasis, caused by Trichomonas vaginalis (T. vaginalis), is a common sexually transmitted disease (STD) that primarily affects the genital and urinary tracts. Trichomoniasis has a worldwide distribution and is also widely endemic in China. In recent years, due to the continuous increase in the proportion of recurrent or persistent vaginal trichomoniasis, the related clinical diagnosis and treatments are facing new challenges. Currently, clinical diagnosis relies on clinical features and laboratory tests. Microscopic examination of vaginal secretions is a commonly used diagnostic method in which active T. vaginalis can be observed under a microscope in vaginal secretions. Although microscopic examination has a high specificity, the sensitivity is only 50 %-60 %. Therefore, the development of more sensitive and accurate clinical diagnostic methods are urgently needed for trichomoniasis. In this study, a clinical diagnosis of T. vaginalis infection was conducted at the Maternal and Child Health Hospital of Hubei province, Xiangyang No.1 People's Hospital, Hunan University of Chinese Medicine, and Jingzhou Central Hospital. Different methods-fluorescent PCR (F-PCR), microscopic examination, sequencing and culture-were employed to diagnose T. vaginalis. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), negative likelihood ratio (NLR), and accuracy of F-PCR are 100 % (98-100 %), 98 % (96 %-99 %), 97 % (93 %-98 %), 100 % (99 %-100 %), 0.00, and 99 % (97 %-100 %), respectively. Compared with traditional methods such as microscopic examination, F-PCR offers superior sensitivity and accuracy. Additionally, F-PCR has a lower probability of false negatives compared with sequencing and culture-based approaches. Compared with methods such as microscopic examination, sequencing, and culture, it significantly reduces the probability of false negatives; and is more sensitive and accurate, making it a powerful diagnostic method for clinical diagnoses.

Chronic kidney disease (CKD) is a growing global health burden characterized by progressive renal dysfunction, high comorbidity rates, and substantial healthcare costs. Digital health technologies, including telemedicine, wearable biosensors, mobile health applications, and AI-driven decision support systems, offer transformative potential for improving CKD management across the care continuum. This review synthesizes current evidence on digital health integration in CKD, highlighting its role in early detection, remote monitoring, patient self-management, and personalized therapeutic interventions. Mechanistically, digital platforms enable real-time tracking of renal biomarkers, blood pressure, and medication adherence while facilitating predictive analytics for risk stratification and disease progression. Integration with electronic health records and interoperability standards further enhances multidisciplinary coordination and data-driven care. Despite promising outcomes in pilot studies and clinical trials, challenges remain in terms of scalability, data privacy, digital literacy, and regulatory compliance. Future directions include the development of interoperable, patient-centric digital ecosystems and validation through multicenter trials. Digital health integration represents a paradigm shift toward proactive, precision nephrology, with the potential to mitigate CKD burden and improve patient outcomes.