Biomarker Insights最新文献

Background: Chronic non-specific low back pain (CNSLBP) is a debilitating condition with unclear underlying mechanisms. The presence of systemic biomarkers associated with inflammation in nonspecific low back pain (NSLBP) has been inconsistently reported primarily through invasive blood sampling.

Objective: This study evaluates the use of saliva as an alternative medium for assessing inflammatory biomarker levels in patients with CNSLBP.

Design: Prospective cross-sectional pilot study.

Methods: Twenty-five patients with CNSLBP and 25 age and sex matched asymptomatic participants were selected according to specific inclusion and exclusion criteria. The primary outcome was determination of the levels of inflammatory biomarkers in unstimulated saliva samples of CNSLBP patients relative to controls using Luminex™ 200 technology. Secondary outcomes were pain, disability and anxiety/stress levels of participants.

Results: In CNSLBP patients, 9 biomarkers interferon γ (IFNγ), interleukin-2 (IL-2), IL-4, IL-5, IL-10, IL-13, IL-12p40, IL-12p70, and tumor necrosis factor α (TNFα) were comparable to controls (P = .25-.94). However, 4 pro-inflammatory mediators were significantly elevated, exhibiting medium to large effect sizes: IL-1β (P = .028, Cohen's d = 1.62), IL-6 (P = .001, d = 1.0), IL-8 (P = .002, d = 0.86), and MCP-1 (P = .001, d = 0.77). Additionally, IL-1Ra levels were significantly higher, though with a small effect size (P = .03, d = 0.43). A significant correlation (P = .02) was observed between VAS pain scores and MCP-1 levels.

Conclusion: Saliva represents a viable medium for assessing key inflammatory biomarkers in patients with chronic non-specific low back pain (CNSLBP). Elevated levels of proinflammatory cytokines, IL-1, IL-6, IL-8, and the nociceptive chemokine MCP-1 were observed in comparison to asymptomatic controls, with MCP-1 showing a positive correlation with self-reported pain intensity. Future studies utilizing unstimulated saliva samples may further investigate changes in inflammatory biomarker levels to monitor treatment outcomes.

Background: Biological knowledge and patient care have been significantly improved by the emergence of big data analysis and -omics sciences, requiring high quality standards for biospecimen and data collection. Biobanks are complex and dynamic units designated to fulfill these needs.

Objectives: The Genoa Vascular Biobank (GTB-VD) is a collaborative network between the IRCCS Ospedale Policlinico San Martino (Centre of Biological Resources), and the University of Genoa (Vascular and Endovascular Surgery Unit; Anatomic Pathology Unit; Laboratory of Clinical and Experimental Vascular Biology). This work describes workflow, ethic and governance requirements, demographic and clinical characteristics of subjects enrolled in the GTB-VD, and the volume of open or endovascular surgical interventions.

Design: The GTB-VD recruits patients undergoing surgical repair for carotid artery stenosis (CS) and abdominal aortic aneurysm (AAA), enrolled on the basis of selection criteria and subdivided for pathology and type of intervention, upon informed consent.

Methods: Biospecimens comprise serum, plasma, whole blood, peripheral blood mononuclear cells, and urine (from AAA only), stored at -80°C; lesions from open surgeries are frozen and formalin fixed paraffin embedded. Samples are associated with donor's clinical data through pseudonymization to prevent patient identification. Data accuracy and sample quality are ensured by harmonized standard operative procedures.

Results: From 2018 to the end of 2023, 442 CS (distinguished into severe-asymptomatic or symptomatic, displaying a ratio of 5:1) and 214 AAA have been collected. CS is more frequently associated with diabetes and peripheral artery diseases, AAA with pulmonary history, and renal function impairment. Open surgery is more used for CS and endovascular for AAA.

Conclusion: The GTB-VD, as organized, represents an "unicum" in our Country; it supports studies to identify molecular targets and biomarkers associated with specific arteriopathy, for developing secondary prevention strategies and minimally invasive, in situ therapies. Collaborative studies and sample sharing are welcome.

Background: Demyelination and remyelination are major issues for scientists dealing with myelin disorders in both clinical and research fields. Despite that, rapid, reliable and convenient tools to monitor myelin changes still lack both in central and peripheral nervous system. Given that myelin is enriched in specific lipids and proteins, it is reasonable they could represent eligible candidates as structural damage biomarkers for this characteristic membrane. Among them, we focused on sphingomyelin (SM) due to the enrichment in myelin and because it is easily measurable in different biological matrices.

Objective: Depicting the roadmap to identify and validate SM dosage as a myelin biomarker useful for pre-clinical and clinical practice.

Design: This study adheres to STROBE guidelines for observational cross-sectional studies on human patients and to ARRIVE guidelines for animal models.

Method: Following the recommendations of the Society for CSF Analysis and Clinical Neurochemistry, we describe the stepwise process to validate SM as a myelin biomarker, starting from the optimization of the fluorescence-based assay and analytical validation in experimental models until clinical and pathological validation in biological fluids of neurological patients.

Results: SM dosage monitors myelination, demyelination, remyelination and even small myelin changes associated to myelin pathology and pharmacological treatments in experimental models. SM is detectable in human biological fluids and informative of myelin damage in the CSF of neurological patients. SM dosage identifies myelin breakdown in the CSF of patients affected by Guillain-Barrè Syndrome (GBS) and Chronic Inflammatory Demyelinating Polyradiculoneuropathy (CIDP), identifying disease activity, axonal from demyelinating variants, and avoiding misdiagnosis.

Conclusion: SM dosage displayed extremely promising real-word performances being able to identify, monitor and stage myelin pathology. Given that it is simple, inexpensive and easily adaptable to routine use in any hospital setting, it might rapidly progress to the implementation and impact on clinical outcomes.

Background: Cancers cause changes in the levels of inflammatory cytokines by inhibiting or promoting their production thus affecting the immune system. The measurement of serum levels of cytokines may be useful in assessing these immunological changes and invariably assessing cancer status.

Objectives: To investigate the effect of imatinib mesylate (glivec^®) on the serum levels of interleukins (IL-6 and IL-10), and C-reactive protein (CRP) in patients with chronic phase chronic myeloid leukaemia (CP-CML).

Design: This prospective cohort study included 26 imatinib naïve CP-CML patients with no other co-morbidities and 26 age and sex-matched healthy controls.

Method: Serum levels of interleukins (IL6 and 10) and CRP were determined using the ELISA method at recruitment for both patients and controls and repeated for the CML patients at 3 months into imatinib therapy.

Results: The mean serum levels of IL-6 and CRP were significantly higher in CP-CML than in the controls at recruitment 439.83 ± 167.52 versus 39.62 ± 10.11 pg/ml, (t = 8.720 P ⩽ .0001), (8.45 ± 2.88 vs 2.86 ± 1.08 mg/l; t = 6.729 P ⩽ .0001) respectively. In contrast, the mean of the IL-10 in the controls (36.63 ± 12.43) was noticed to be significantly higher than the patients (22.88 ± 4.76 vs 36.63 ± 12.43 pg/ml; t = -3.851 P = .003). Interestingly, there was a significant drop in the serum levels of IL-6 (439.83 ± 167.52 vs 46.85 ± 14.48 pg/ml, (t = 8.055 P ⩽ .0001) and CRP (8.45 ± 2.88 mg/l vs 4.24 ± 1.57; t = 4.305 P = .0001) in the CML subjects 3 months into imatinib therapy. Only IL-10 had a non-significant drop in the CML subjects after 3 months of imatinib therapy. Method validation of these biomarkers was done using the Receiver operating characteristic (ROC) curve which revealed an area under the curve (AUC) of 1.000 for both IL-6 and CRP and 0.152 for IL-10.

Conclusion: The study has concluded that treatment naïve CML is associated with a significant elevation of pro-inflammatory cytokines (IL-6 and CRP) and treatment with imatinib led to a significant decline in the serum levels of these markers suggesting that IL-6 and CRP could be useful as adjunct in the monitoring of CML treatment.

Background: Bacterial infections are often an overlooked factor in female infertility. Staphylococcus aureus has been identified as the predominant vaginal pathogen in infertile women, with a prevalence of 57.33%. Previous studies showed that S. aureus induces infertility in mice by sperm impairment, suggesting its asymptomatic vaginal colonization creates a hostile environment for sperm. While sperm immobilization factor (SIF) from culture supernatant has been identified, its production within host's environment remained unexplored.

Objective: To unveil S. aureus-derived signature protein(s) in vaginal lavage fluid (VLf).

Design: Mass spectrometry combined with experimental studies.

Methods: VLf was obtained from female mice administered either with sperm immobilizing S. aureus (test group) or PBS alone (control group) and analyzed using nano-LC-MS/MS, gel filtration chromatography, SDS-PAGE, functional assays, and in silico studies.

Results: Nano-LC-MS/MS yielded 5 distinct bacterial proteins in test group and no bacterial protein in control. Elution profile of test VLf revealed a single peak and indicated 1 protein band (~36 kDa) using SDS-PAGE that aligned with GMP reductase. VLf-protein showed impairment of sperm motility and viability in concentration-dependent manner and disrupted sperm morphology. Binding studies using FITC-labeled VLf-protein depicted presence of green fluorescence over entire surface of mouse spermatozoa. These results were akin to SIF, already isolated and characterized in our laboratory, from culture supernatant of S. aureus, causing sperm impairment and hence, designated as vaginal lavage fluid-derived sperm immobilization factor (VLf-SIF). Through in silico analysis, superimposition of VLf-SIF and SIF, already known to show sequence homology to cysteine-tRNA ligase, revealed close structural alignment. Molecular docking analysis depicted energetically favorable binding between VLf-SIF and spermatozoa surface protein (Heat shock-related 70 kDa protein 2).

Conclusion: This study provides novel evidence of sperm-impairing S. aureus signature proteins as key mediators of bacterial-induced infertility, paving way for diagnostic, and therapeutic advancements.

With an aging population, the demand for sensitive and specific biomarkers to assess bone turnover has surged. Bone turnover involves 2 key processes: bone formation, during which Type I procollagen is cleaved into Type I collagen and subsequently mineralized into bone, and bone resorption, during which Type I collagen is demineralized and degraded into peptides by cathepsin K. To identify biomarkers that accurately reflect these processes, extensive efforts have been made to characterize the peptides generated during both formation and resorption. Over the years, numerous biomarkers have been discovered for various disorders. However, despite their clinical utility, many of these markers lack specificity. This is due to factors such as the degradation of trimers into monomers, the coexistence of multiple peptide species arising from the unpredictable cleavage of Type I collagen/procollagen by cathepsin K and metalloproteinases, and the lack of assay standardization. Standardization is further hindered by the incomplete characterization of many of these peptides. For accurate assay development, a gold-standard technique like LC-MS/MS is essential, requiring full peptide characterization during method development. This review aims to present recent advances in the characterization of Type I collagen-derived peptides, providing a foundation for improved biomarker standardization and application in clinical practice.

Background: Demographic, health history, and lifestyle factors have been associated with prognosis of colorectal cancer (CRC), but mechanisms underlying these associations remain poorly understood. A compelling mechanism involves changes in expression of tumor markers that influence treatment outcomes, such as secreted protein acidic and rich in cysteine (SPARC), lower levels of which have previously been associated with poorer CRC prognosis.

Objective: We explored the association of factors that have been previously associated with CRC prognosis with expression of SPARC in tumor tissues.

Design: We conducted a prospective evaluation of 50 participants of a longitudinal cohort study that went on to develop CRC.

Methods: Tumor and normal tissue cores were taken from formalin-fixed paraffin-embedded (FFPE) blocks of incident CRC cases and were used to create tissue microarrays (TMAs). Slides created from the TMAs were stained with SPARC antibodies and analyzed to calculate H-scores for both epithelial and non-epithelial components of tumor and normal tissues. H-scores were ln-transformed and analyzed in association with demographic, lifestyle, and health history factors assessed before cancer diagnosis using linear regression models.

Results: In CRC tumor epithelium, smoking was associated with a 0.53-fold lower level of SPARC expression (P = .054). Higher income was associated with a 1.33-fold greater level of SPARC expression in tumor non-epithelial tissue (P = .041). Higher cancer stage was associated with a 0.74-fold lower level of non-epithelial tumor SPARC expression (P = .040). In the epithelial component of normal colorectal tissues, higher fruit consumption was associated with a 2.74-fold greater SPARC H-score (P = .002).

Conclusions: The associations we observed for smoking, income, and cancer stage with SPARC in tumor tissue are consistent with previously established associations of these factors with CRC prognosis. Larger studies with prognostic data are needed, but our results suggest that differences in SPARC expression may contribute to previously observed impacts of various factors on CRC prognosis.

Background: Noninvasive and cost-effective markers are needed to replace esophagogastroduodenoscopy in the screening for severe esophagogastric varices (EGVs) and portal hypertensive gastropathy (PHG).

Objective: This study evaluated the performances of several commonly used fibrosis markers in assessing EGVs and PHG in cirrhosis patients.

Design: Retrospective cohort study.

Methods: A series of 323 patients with cirrhosis were consecutively enrolled and endoscopically followed up until variceal eradication was achieved. The Fibrosis-4 (FIB-4) score, albumin-bilirubin (ALBI) index, aspartate aminotransferase (AST)-to-alanine aminotransferase (ALT) ratio (AAR), AST-to-platelet ratio index (APRI), gamma-glutamyl transpeptidase-to-platelet ratio (GPR), and Lok score were calculated for each patient upon first admission. The performances of these markers in assessing EGVs and PHG were determined.

Results: In the screening for clinically relevant esophageal varices (CREVs), none of the markers showed a significant ability to differentiate CREVs from non-CREVs (P > .05). The AAR (area under the curve (AUC): 0.581, sensitivity: 52.0%, specificity: 66.1%, P = .033) and the GPR (AUC = 0.596, sensitivity: 64.0%, specificity: 50.0%, P = .033) fairly differentiated clinically relevant gastric varices (CRGVs) from non-CRGVs patients. Moreover, no correlation was noted between PHG and CREVs (r = .016, P = .778) or between PHG and CRGVs (r = -.024, P = .666). Furthermore, no difference in the severity of PHG before and after variceal eradication was detected (P = .224).

Conclusion: The studied markers revealed poor to no ability to assess EGVs or PHG. Hence, they cannot be used to substitute EGD in the screening for EGVs. Furthermore, endoscopic eradication of EGVs did not affect the severity of PHG.

Background: Procalcitonin (PCT) is recognized as an inflammatory biomarker, often elevated in COVID-19 pneumonia alongside other biomarkers. Understanding its association with severe outcomes and comparing its predictive ability with other biomarkers is crucial for clinical management.

Objectives: This retrospective multicenter observational study aimed to investigate the association between PCT levels and adverse outcomes in hospitalized COVID-19 patients. Additionally, it sought to compare the predictive performance of various biomarkers.

Design: The study analyzed data from the Society of Critical Care Medicine (SCCM) Viral Infection and Respiratory Illness Universal Study (VIRUS) registry, comprising COVID-19 patients hospitalized across multiple Mayo Clinic sites between March 2020 and June 2022.

Methods: A total of 7851 adult COVID-19 patients were included. Patients were categorized into 6 groups based on the worst WHO ordinal scale. Multivariate models were constructed using peak biomarker levels within 72 hours of admission, adjusted for confounders.

Results: Elevated PCT levels were independently associated with increased odds of adverse outcomes, including ICU admission (adjusted odds ratio [aOR] 1.32, 95%CI 1.27-1.38), IMV requirement (aOR 1.35, 95%CI: 1.28-1.42), and in-hospital mortality (aOR 1.30, 95%CI: 1.22-1.37). A 3.48-fold increase in IMV requirement and 3.55 times increase in in-hospital mortality were noted with peak PCT ⩾ 0.25 ng/ml. Similar associations were observed with other biomarkers like NLR (AUC 0.730), CRP, IL-6, LDH (AUC 0.800), and D-dimer (AUC 0.719). Models incorporating NLR, LDH, D-dimer, and PCT demonstrated the highest predictive accuracy, with a combined model exhibiting an area under the curve (AUC) of 0.826 (95%CI 0.803-0.849).

Conclusions: Higher PCT levels were significantly linked to worse outcomes in COVID-19 patients, emphasizing its potential as a prognostic marker. Biomarker-based predictive models, particularly those including PCT, showed promising utility for risk assessment and clinical decision-making. Further prospective studies are warranted to validate these findings on a larger scale.

Background: Prostate cancer (PCa) patients with biochemical recurrence (BCR) following radical prostatectomy or radiation therapy often present with very low prostate-specific antigen (PSA) levels (⩽0.5 ng/mL). Accurate detection of recurrence at such low levels is crucial for guiding treatment decisions.

Objectives: To assess the diagnostic efficacy of [¹⁸F]PSMA-1007 PET/MR (PSMA-PETMR) in detecting BCR of PCa in patients with very low PSA levels.

Design: A prospective study conducted between May 2021 and January 2023, with data subsequently analyzed retrospectively after a 2-year follow-up.

Methods: The cohort comprised 157 PCa patients with BCR, of whom 52 had PSA levels ⩽ 0.5 ng/mL and underwent PSMA-PETMR imaging. The imaging protocol incorporated multiparametric MRI (mpMRI) and PET acquisitions, with lesion classification following PSMA-RADS version 1.0. Detection rates of recurrent lesions, including local recurrence, lymph node metastasis, and skeletal metastasis, were evaluated.

Results: PSMA-PETMR exhibited a 63.5% detection rate for recurrent PCa at low PSA levels, surpassing traditional diagnostic methods. Thirty-four local recurrences, 12 metastatic lymph nodes, and 4 skeletal metastases were identified. Follow-up imaging enhanced the detection rate to 73.1% by reclassifying initially equivocal findings. PSMA-PETMR influenced clinical decision-making in 17% of patients by facilitating personalized treatment strategies.

Conclusion: PSMA-PETMR significantly improves the detection of recurrent PCa in patients with very low PSA levels, offering precise lesion localization and supporting personalized treatment approaches. Further studies are needed to optimize its clinical use and validate its long-term benefits.