Potential Diagnostic Utility Research Articles

Characteristic TARC/CCL17 expression in the salivary gland of IgG4-related disease: potential diagnostic utility and insights into pathogenesis

Immunoglobulin G4-related disease (IgG4-RD) is a chronic inflammatory condition of unknown etiology characterized by lymphocytic infiltration, fibrosis, and infiltration of IgG4-positive plasma cells. It affects various organs, including the pancreas and salivary glands. Immunological abnormalities are suspected to play a role in its pathogenesis, and there is an epidemiological link to allergic conditions and type 2 inflammation. This study focused on the expression of thymus and activation-regulated chemokine (TARC)/CCL17, which is involved in the migration of T helper 2 and/or regulatory T cells, in salivary gland tissues of patients with IgG4-RD. We analyzed 60 salivary gland biopsy samples obtained from patients at Sapporo Medical University Hospital between 2015 and 2020. Immunohistochemical analysis revealed TARC/CCL17 positivity in 87.2% of histologically confirmed IgG4-RD cases and negativity in 84.6% of histologically unconfirmed but clinically suspected IgG4-RD cases. There was a significant correlation between histologically confirmed IgG4-RD and TARC/CCL17 expression, suggesting its potential diagnostic utility and possible involvement in the pathogenesis of IgG4-RD.

Decreased serum PF4 levels correlate with cognitive decline and CSF biomarkers in Alzheimer's disease in a Chinese cohort.

Platelet factor 4 (PF4), a chemotactic factor secreted from the α-granules of platelets, has recently been proved to mitigate neuroinflammation and improve aging-related cognition decline, which may be involved in Alzheimer's disease (AD). This study aims to investigate the alterations of serum PF4 levels in AD, the correlation between serum PF4 and β-amyloid (Aβ) and tau levels in cerebrospinal fluid (CSF), and the potential diagnostic utility of PF4 in AD. A cross-sectional study was conducted involving 38 amyloid-positive AD patients and 50 cognitively normal controls. The levels of serum PF4 were detected using the Human CXCL4/PF4 enzyme-linked immunosorbent assay (ELISA) Kit. The levels of CSF Aβ42, Aβ40, p-tau181, and t-tau were measured on the fully-automated Lumipulse G1200 platform via commercially available kits. The levels of serum PF4 were significantly decreased in AD patients (5163.51 (3198.24-6301.15) vs. 5859.29 (4126.06-8006.70), Z=-2.30, P=0.021). The negative correlation between AD diagnosis (β=-1972.292, P=0.009) and PF4 levels retained after the adjustments of age, sex, APOE ε4 status, platelet count, platelet distribution width (PDW), and comorbidity of dyslipidemia in the multiple linear regression analysis. Further analysis showed that serum PF4 levels were positively correlated with CSF Aβ42 levels and Mini-Mental State Examination (MMSE) scores, and negatively correlated with CSF t-tau levels. Besides, the area under the curve (AUC) of serum PF4 for AD (AUC=0.6437, P=0.022) was comparable to that of CSF Aβ40 (AUC=0.6400) yet lower than those of CSF Aβ42, ptau181, and t-tau. The AUC slightly increased when combining serum PF4 with other CSF AD biomarkers separately. The serum levels of PF4 were decreased in AD patients and were significantly correlated with the cognitive function and CSF levels of Aβ42 and t-tau. PF4 may become a promising anti-aging and therapeutic target for AD, which is worthy of further study.

Clinical utility of plasma p-tau217 in identifying abnormal brain amyloid burden in an Asian cohort with high prevalence of concomitant cerebrovascular disease.

Using an Asian cohort with high prevalence of concomitant cerebrovascular disease (CeVD), we evaluated the performance of a plasma immunoassay for tau phosphorylated at threonine 217 (p-tau217) in detecting amyloid beta positivity (Aβ+) on positron emission tomography and cognitive decline, based on a three-range reference, which stratified patients into low-, intermediate-, and high-risk groups for Aβ+. Brain amyloid status (Aβ- [n=142] vs Aβ+ [n=73]) on amyloid PET scans was assessed along with the plasma ALZpath p-tau217 assay to derive three-range reference points for PET Aβ+ based on 90% sensitivity (lower threshold) and 90% specificity (upper threshold). Plasma p-tau217 (area under the curve [AUC]=0.923) outperformed routine clinical assessments (AUC=0.760-0.819; p ≤ 0.003) and other plasma biomarkers (AUC=0.817-0.834; p < 0.001). The high-risk group showed significantly higher rates of cognitive decline than the low-risk group. Risk stratification for PET Aβ+ based on a plasma p-tau217 assay demonstrated potential diagnostic and prognostic utility in an Asian cohort with concomitant CeVD. The utility of plasma p-tau217 to detect brain amyloid beta pathology (Aβ+) was studied in an Asian cohort with concomitant cerebrovascular disease Plasma tau phosphorylated at threonine 217 (p-tau217) showed superior utility in detecting Aβ+ compared to neuroimaging measures, clinical workup, or other blood biomarkers including p-tau181, glial fibrillary protein (GFAP), and Aβ Higher plasma p-tau217 correlated with faster cognitive decline Plasma p-tau217 shows promise as an Alzheimer's disease (AD) diagnostic and prognostic biomarker in diverse populations.

The Role of Positron Emission Tomography Imaging in Breast Implant Illness.

Explantation often alleviates symptoms in women with breast implant illness. However, persistent complaints in some cases may be linked to persistent silicone-induced inflammation from residual silicone particles. Positron emission tomography (PET) imaging could potentially detect this inflammation. This case series describes the PET findings in women with ongoing symptoms after explantation. A retrospective review was performed of cases from the silicone outpatient clinic at the Amsterdam University Medical Centers, the Netherlands. All women underwent PET imaging due to persistent systemic symptoms after explantation (n = 17) or replacement (n = 1). Before PET imaging, silicone deposits were demonstrated in all 18 cases using ultrasound or magnetic resonance imaging. PET imaging revealed varying fluorodeoxyglucose avidity in axillary, parasternal, mediastinal, cervical, or supraclavicular lymph nodes and extranodal sites in all patients, up to 11 years after explantation. The median implantation time was 17 years, the average number of implant sets was 2, and the median time from explantation to PET was 2 years. In cases where biopsy was performed, silicone lymphadenitis with characteristic foreign body reaction was confirmed. The PET findings suggest that silicone residues can provoke inflammation even years after explantation. However, not all women with silicone residues may exhibit fluorodeoxyglucose-positive PET scans, indicating variability in susceptibility to silicone-induced inflammation. PET imaging may be a useful diagnostic tool for detecting silicone-induced inflammation in patients with persistent complaints after explantation. However, given inherent limitations, further research is warranted to fully assess its potential diagnostic utility in breast implant illness.

YKL-40 levels in cerebrospinal fluid serve as a diagnostic biomarker for post-neurosurgical bacterial meningitis in patients with stroke.

Previous studies have reported that YKL-40 can serve as a biomarker of infectious diseases and different stroke types. However, evidence supporting its role in diagnosing post-neurosurgical bacterial meningitis (PNBM) remains lacking. A total of 110 patients with stroke who underwent neurosurgical treatment were recruited. Among these, 36 were diagnosed with PNBM based on the results of bacterial culture/Gram staining or cerebrospinal fluid (CSF) characteristics. CSF levels of YKL-40 and other biomarkers with potential diagnostic utility for PNBM were statistically analysed using univariate and logistic regression models. Receiver operating characteristic (ROC) analysis was performed to investigate diagnostic efficiency. According to univariate analysis, CSF levels of glucose, total protein, white blood cells, polymorphocytes, and YKL-40, as well as the CSF-to-blood glucose ratio, were significantly different between the PNBM and non-PNBM groups. Logistic regression analysis revealed that glucose (p=0.026), total protein (p=0.028), and YKL-40 (p=0.006) levels in the CSF may have independent diagnostic utility for PNBM. Among the three biomarkers, CSF-to-blood glucose (area under the receiver operating characteristic curve [AUC] 0.9208) and YKL-40 (AUC 0.9587) demonstrated strong diagnostic utility. CSF levels of YKL-40, glucose, and total protein played independent roles in the diagnosis of PNBM in patients with stroke.

Assessment of Trace elements (Zinc, Copper) in Iraqi Patients with Chronic Kidney Disease on Dialysis

Background: In the context of insufficient healthcare accessibility for a substantial segment of the global populace, chronic kidney disease (CKD) has emerged as a significant contributor to mortality, ranked subsequent to cardiovascular disease and neoplasia, demonstrating a notable increase over the preceding two decades. This rising burden of CKD is further exacerbated by a deficient understanding of its underlying pathogenesis. Therefore, it was important to study all the variables in the human body that contribute to monitoring the patient's condition or preventing the patient's condition from deteriorating. In this study, we highlighted some trace elements (zinc and copper), and monitored the change in these elements between patients and healthy people. We indicated the possibility of monitoring the results of tests for these elements to predict the condition of patients from the first stage to the final stages of chronic renal failure patients on dialysis. purpose: To assess the trace elements (zinc, copper) in Chronic kidney disease patients compared with healthy control. Methods: A total forty (40) patient with chronic kidney disease were studied among them twenty (20) female and twenty (20) male patients and forty (40) healthy as control of them were eighteen (18) females and twenty-two (22) males. Their ages (patients and controls) were ranged from (18 – 70). Blood samples were collected from patients and control to evaluate serum levels of serum trace elements (zinc, copper) and evaluate the routine markers (urea, creatinine, Alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase) and this was done by spectrophotometry. Results The Zinc levels in the patients (80.69±51.30 μg/dl) were lower than in the control group (97.42±38.61 μg/dl) (p=0.10773) and The Copper levels in the patients are significantly higher (116.33±71.20 μg/dl) than in the control group (74.77±46.16 μg/dl) (p=0.00304). while The Alanine Aminotransferase and Aspartate Aminotransferase levels in the patients are lower than in the control group, but the difference is not statistically significant for ALT (p=0.13235) and is significant for AST (p=0.02982). the Alkaline Phosphatase, Urea, and Serum Creatinine levels in the patients were all significantly higher than in the control group (p&lt;0.00001 for all). Conclusion: A notable elevation in alkaline phosphatase and copper levels is observed in chronic kidney disease patients, alongside a modest decrease in serum zinc in those with end-stage chronic kidney disease compared to healthy individuals, indicating these biomarkers' potential diagnostic utility in advanced stages of the disease. The correlation between AST/ALT and CKD progression highlights the impact of liver enzyme alterations during chronic kidney disease.

PRAME Staining of Adnexal Lesions and Common Skin Cancer Types: Biomarker with Potential Diagnostic Utility.

PRAME (PReferentially expressed Antigen in MElanoma) is a tumor-associated antigen first identified in tumor-reactive T-cell clones derived from a patient with metastatic melanoma. Immunohistochemistry (IHC) for PRAME is useful for diagnostic purposes to support a suspected diagnosis of melanoma. Anecdotally, PRAME has been observed to stain sebaceous units in glands in background skin. We examined the expression of PRAME in adnexal lesions and common skin cancers to determine whether it is of potential diagnostic utility in supporting the differentiation between sebaceous and non-sebaceous lesions. IRB approval from Mount Sinai Medical Center (MSMC) was obtained. This is a single-center retrospective cohort analysis over a ten-year period (1 January 2012, and 31 December 2023). We used the pathological database of skin lesions, including sebaceous, sweat gland, and follicular lesions, in addition to basal cell carcinomas (BCCs) and squamous cell carcinomas (SCCs), from 81 patients who underwent shave/punch biopsies or surgical excisions. We evaluated the IHC staining percentage positivity and intensity for PRAME. Staining intensity was subcategorized into negative, weak, moderate, and strong, whereas expression percentage positivity was subcategorized into 0%, 1-25%, 26-50%, 51-75%, and 76-100%. Most sebaceous versus non-sebaceous lesions exhibited cytoplasmic staining of moderate to strong intensity in >75% of cells. PRAME has a sensitivity and specificity of 100.0% and 86.7%, respectively, to support distinguishing between sebaceous and non-sebaceous adnexal lesions (regardless of whether they are benign or malignant). BCCs and SCCs showed weak to moderate nuclear staining for PRAME in >75% of cells. None of the 13 lesions of hair follicle origin showed any staining. A total of 26 of the 32 lesions of sweat gland origin were negative while 6 (18.75%) showed positive staining. In conclusion, we confirm the potential utility of PRAME for supporting the distinction between sebaceous and non-sebaceous adnexal lesions on one hand, and on the other, distinguishing BCC and SCC that may show nuclear staining from sebaceous carcinoma that shows cytoplasmic staining.

Exploring the Role of Chemokine-Related Gene Deregulation and Immune Infiltration in Ischemic Stroke: Insights into CXCL16 and SEMA3E as Potential Biomarkers.

Ischemic stroke is a leading cause of mortality and disability globally. Understanding the role of chemokine-related differently expressed genes (CDGs) in ischemic stroke pathophysiology is essential for advancing diagnostic and therapeutic strategies. We conducted comprehensive analyses using the GSE16561 dataset: chemokine pathway enrichment via GSVA, differential expression of 12 CDGs, Pearson correlation, and functional enrichment analyses (GO and KEGG). Machine learning algorithms were employed to develop diagnostic models, evaluated using ROC curve analysis. A nomogram was constructed and validated with independent datasets (GSE58294). Gene set enrichment analysis (GSEA) and immuno-infiltration analysis were also performed. Chemokine pathway scores were significantly elevated in ischemic stroke, indicating their potential involvement. Logistic regression emerged as the most effective diagnostic model, with CXCL16 and SEMA3E as significant biomarkers. The nomogram exhibited high discriminatory ability (AUC = 0.964), well-calibrated predictions, and clinical utility across datasets. GSEA highlighted key biological pathways associated with CXCL16 and SEMA3E. Immuno-infiltration analysis revealed significant differences in immune cell infiltration between control and ischemic stroke groups, with distinct correlations between CXCL16 and SEMA3E expression and immune cell populations. This study highlights the deregulation of CDGs in ischemic stroke and their implications in critical biological processes. CXCL16 and SEMA3E are identified as key biomarkers with potential diagnostic utility. Insights from gene set enrichment and immuno-infiltration analyses provide mechanistic understanding, suggesting novel therapeutic targets and enhancing clinical decision-making in ischemic stroke management.

Association of Plasma Alzheimer’s Disease Biomarkers with White Matter Hyperintensity Volume in Diverse Populations: A HABS‐HD Study

AbstractBackgroundOlder African American (AA) and Hispanic American (HA) adults have a higher risk for Alzheimer’s disease (AD) than older non‐Hispanic white (NHW) adults. Cerebrovascular disease reflected by white matter hyperintensity (WMH) lesions on magnetic resonance imaging (MRI) may influence ADRD risk in these groups. Amyloid and tau PET data from studies of mostly NHW participants show a relationship between AD pathology and WMH lesions. Using a diverse cohort, this study investigates relationships between plasma AD biomarkers and WMH volume.MethodWe analyzed plasma AD biomarkers, WMH volume, and self‐reported participant race (NHW, HA, AA) obtained during baseline visits for the Health and Aging Brain Study: Health Disparities. Plasma AD biomarkers included amyloid‐beta40 (Aβ40), amyloid‐beta42 (Aβ42), total tau (T‐tau), phosphorylated tau181 (P‐tau181), and neurofilament light (NfL) from single‐molecule enzyme‐linked immunosorbent assays. From self‐reported race and baseline global Clinical Dementia Rating scores (normal=0, impaired≥0.5), we created race/cognitive groups (white/normal, white/impaired, AA/normal, AA/impaired, HA/normal, HA/impaired). Using ANOVA and chi‐square, we compared baseline characteristics, plasma AD biomarkers, and WMH volume in the groups. We tested associations between plasma AD biomarkers and WMH volume with linear regression adjusted for age, education, and sex.ResultData from 3,035 participants was analyzed (mean age: 65.2; female: 62.3%; impaired: 20.1%). AA persons were younger (NHW: 68.6, AA: 62.9, HA: 62.3. p&lt;.0001) and had the highest WMH volume (NHW: 4.16, AA: 18.4, HA: 3.18, p&lt;.0001). AA and HA participants had the lowest Aβ42 (NHW:10.9, AA:10.2, HA:10.1, p&lt;.0001), while HA persons had the lowest Aβ40 (NHW: 223, AA: 220, HA: 207, p=0.0003). NHW persons had the highest P‐tau181 (NHW: 2.38, AA: 1.95, HA: 1.92, p&lt;.0001) and NfL (NHW:19.1, AA: 12.3, HA: 17.1, p&lt;.0001). Regression showed higher WMH volume was associated with higher Aβ40 (β=.02, p=.02), P‐tau181 (β=.29, p=.01), and T‐tau (β=1.55, p=.0002) in the HA/impaired group.ConclusionOur results show associations between plasma AD biomarkers and WMH volume in cognitively impaired HA adults who also have low WMH burden compared to other groups. Further studies to discern potential mechanisms and the potential diagnostic prognostic utility of plasma AD biomarkers on AD risk are needed.

Truncated Albumins as Novel Surrogate Biomarkers in Diabetes Therapy: Epiphenomena and Potential Clinical Applications.

Aims/Hypothesis: Among various albumin posttranslational modifications (PTMs), N- and C-terminal truncations (HSA-DA and HSA-L) have also shown biomarker potential in disease states. We examined albumin truncation longitudinal trends and correlations during diabetes therapy toward possible future clinical applications. Methods: In a preliminary longitudinal therapy investigation, mass spectrometry was employed to track PTMs of human serum albumin (HSA), including glycation (GA), cysteinylation (CA or HNA1; reversible), di/trioxidation (OA or HNA2; irreversible), and truncation (TA). These modifications were correlated with ongoing therapy in four distinct subject groups: type 1 diabetes (T1DM), type 2 diabetes (T2DM), prediabetes-obesity (PDOB), and healthy controls (NORM), observed over a follow-up period extending up to 280 days. Results: Diabetes was associated with significant reduction ("deficiency") of measured albumin truncations (For HSA-DA: T2DM = 0.32 ± 0.3%, p = 2E-08; T1DM = 1.02 ± 0.4%, p = 3E-05; PDOB = 1.61 ± 0.2%, p = 0.004; compared to NORM = 2.08 ± 0.2%). Albumin truncation reduction was more striking in T2DM (HSA-DA: T2DM vs. T1DM: p = 0.004). Improvements in glycemic control and decrease of albumin glycation during diabetes therapy were associated with concomitant increase of albumin truncations toward the "healthy" normal ranges, and vice versa ("mirror image" trends). Accordingly, albumin truncation correlated inversely with albumin glycation (HSA-DA vs. GA: R = -0.53, p = 1E-09). Conclusions: The "epiphenomenon" of albumin truncation (reflecting the severity of mean hyperglycemia and also insulin resistance) can possibly provide novel, sensitive, and complementary biomarkers (e.g., via simpler HSA-DA peptide fragment immunoassays) to monitor efficacy of diabetes therapy and also progression from "healthy" to prediabetes and type 2 diabetes, highlighting potential diagnostic and prognostic utility in clinical diabetes care.

Diagnostic Value of Maximal Systolic Acceleration Measurements in the Follow-up of Complex Endovascular Aortic Repair: Illustration of a Concept.

After complex endovascular aortic repair (cEVAR), long-term surveillance is advocated to monitor for potential (stent-related) complications. Although various imaging modalities are used, computed tomography angiography remains the standard in current clinical practice worldwide. However, radiopaque markers can cause metal artifacts and scattering, hampering assessment of patency of side branches. The maximal systolic acceleration (ACCmax) is a relatively new duplex ultrasound (DUS) parameter measured distal to a stenosis, avoiding in-stent assessment and scattering. The aim of this article is to illustrate the potential diagnostic utility of the ACCmax after cEVAR and visceral artery stenting in general. The ACCmax is measured at the maximal slope of the upstroke during the systolic phase in arterial flow. By manually connecting 2 points, a tangent line is created in which the maximal acceleration is automatically calculated and expressed in m/s2. A higher value reflects better arterial perfusion proximal to its measurement point. One measurement of a visceral artery takes about 5 minutes. ACCmax measurements can be a useful addition during DUS follow-up after cEVAR or visceral artery stenting, possibly decreasing radiation exposure, nephrotoxicity, and health care costs. Long-term surveillance after complex endovascular aortic repair (cEVAR) is necessary to monitor for potential (stent-related) complications. Although CTA is the most common modality for imaging, metal artifacts and scattering can hamper the assessment of stent patency. The maximal systolic acceleration (ACCmax) is a duplex ultrasound based parameter that reflects arterial perfusion proximal to its measurement point. Due to its noninvasive nature, it can be a useful addition during follow-up after cEVAR or visceral artery stenting, possibly decreasing radiation exposure, nephrotoxicity and health care costs as well.

Comprehensive Analysis and Experimental Validation of HEPACAM2 as a Potential Prognosis Biomarker and Immunotherapy Target in Colorectal Cancer.

The role of HEPACAM family member 2 (HEPACAM2) is unclear in colorectal cancer (CRC). The objective of this study was to perform an extensive examination of HEPACAM2 and validate it experimentally in CRC. This study investigated the significance of HEPACAM2 in CRC and its potential diagnostic utility utilizing data from the Cancer Genome Atlas (TCGA) database. Additionally, the study examined potential regulatory networks involving HEPACAM2, including its associations with immune infiltration, immune checkpoint genes, tumor mutational burden (TMB), microsatellite instability (MSI), mRNA expression-based stemness index (mRNAsi), and drug sensitivity in CRC. The expression of HEPACAM2 was further validated using the GSE89076 dataset, and quantitative reverse transcription PCR (qRT-PCR) was employed to confirm HEPACAM2 expression levels in six pairs of CRC tissue samples. HEPACAM2 exhibited abnormal expression patterns in various types of cancer, including CRC. A decrease in HEPACAM2 expression levels in CRC was found to be significantly correlated with the T stage (p < 0.001). Reduced HEPACAM2 expression in CRC patients was also linked to poorer overall survival (OS) (p = 0.007). The expression levels of HEPACAM2 in CRC patients were identified as an independent prognostic factor (p = 0.016). Furthermore, HEPACAM2 was associated with TCF-dependent signaling in response to WNT, G2/M checkpoints, and other pathways. The expression of HEPACAM2 in CRC was found to be associated with immune infiltration, immune checkpoint genes, TMB / MSI, and mRNAsi. Additionally, the expression of HEPACAM2 in CRC was significantly and inversely correlated with the drug sensitivities to gw772405x and 6-phenyl-6h-indeno[1,2-c]isoquinoline-5,11-dione. qRT-PCR confirmed that the expression level of HEPACAM2 was found to be lowly expressed in CRC tissues. These findings suggest that HEPACAM2 may serve as a potential prognostic biomarker and immunotherapeutic target for CRC patients.

Exploring the molecular landscape of cutaneous mixed tumors characterized by TRPS1::PLAG1 gene fusion.

The histological similarities between pleomorphic adenomas (PAs) and cutaneous mixed tumors (CMTs) found in certain facial regions can create a diagnostic challenge. Molecular findings reveal common genetic profiles, particularly PLAG1 rearrangements in both PA and CMT. Although molecular distinctions have received limited attention, our observations indicate multiple cases of CMTs carrying the TRPS1::PLAG1 fusion. This clinical experience has driven our investigation into the potential diagnostic utility of TRPS1::PLAG1 fusions for determining tumor origin. Two cohorts consisting of 46 cases of CMT and 45 cases of PA of the salivary glands were obtained from French institutions and reviewed by specialists in each subspecialty. RNA sequencing analysis was conducted to identify the molecular features of cases harboring PLAG1. Clinical, pathological, and molecular data were collected. In this study, cases of CMT exhibited recurrent gene fusions, primarily TRPS1::PLAG1 (74%). These tumors shared characteristic histological features, including tubuloductal differentiation in 55% of cases and squamous metaplasia in varying proportions. In contrast, cases of PA had gene fusions involving PLAG1 with various gene partners, with only one case in which TRPS1::PLAG1 was identified. This disparity was also observed at the transcriptomic level between TRPS1::PLAG1 CMTs and other tumors. However, TRPS1 immunostaining did not correlate with TRPS1::PLAG1 fusion. In conclusion, we report that recurrent TRPS1::PLAG1 fusion CMTs exhibit similar characteristic histological features, including tubuloductal differentiation that is associated with squamous metaplasia in around half of cases. Detection of this fusion could be valuable in correctly identifying the origin of these tumors. © 2024 The Pathological Society of Great Britain and Ireland.

New strategies to classify canine pleural effusions and the diagnostic value of acute phase proteins, amylase, and adenosine deaminase in pleural exudates.

In dogs, simplified Light's criteria can discriminate transudates from exudates. Other tests used in human medicine are pleural effusion cholesterol (CHOLPE) and butyrylcholinesterase [BChEPE], the pleural effusion/serum ratio of these analytes (CHOLratio and BChEratio), and the serum albumin minus pleural effusion albumin gradient (SEAG). We aimed to assess the diagnostic accuracies of different biomarkers in dogs with pleural effusion in differentiating exudates from transudates. Secondarily, we evaluated the potential diagnostic utility of pleural effusion acute phase proteins, amylase, and adenosine deaminase in discriminating causes of exudative effusions. Cross-sectional study including 68 client-owned dogs with pleural effusion. There were 48 exudates (10 septic, 16 neoplastic, 9 hemorrhagic, and 13 classified as other exudates) and 20 transudates. All the variables analyzed, except SEAG, were significantly different between exudates and transudates. Using the cut-off values adopted in human literature, accuracies for CHOLPE, CHOLratio, BChEPE, and BChEratio were between 82.35% and 85.29%; all values were significantly lower compared with the previously published simplified Light's criteria accuracy (i.e., 98%, p < .001 for all comparisons). We found the accuracy of the simplified Light's criteria to be similar to what has been previously reported (95.59%, p = .238). Paraoxonase-1 (PON-1PE) activity and the pleural effusion/serum paraoxonase-1 ratio (PON-1ratio) were significantly lower in exudative neoplastic effusions than in exudative hemorrhagic (p = .004 and p = .001) and septic (p = .004 and p < .001) effusions. Simplified Light's criteria were the best method for discriminating transudates from exudates, and a low PON-1PE activity and PON-1ratio in exudative effusions may suggest an underlying neoplasia.

Exploiting the role of CSF NfL, CHIT1, and miR-181b as potential diagnostic and prognostic biomarkers for ALS

Amyotrophic lateral sclerosis (ALS) is a rare neurodegenerative disorder characterized by relentless and progressive loss of motor neurons. A molecular diagnosis, supported by the identification of specific biomarkers, might promote the definition of multiple biological subtypes of ALS, improving patient stratification and providing prognostic information. Here, we investigated the levels of neurofilament light chain (NfL), chitotriosidase (CHIT1) and microRNA-181b (miR-181b) in the cerebrospinal fluid (CSF) of ALS subjects (N = 210) as well as neurologically healthy and neurological disease controls (N = 218, including N = 74 with other neurodegenerative diseases) from a large European multicentric cohort, evaluating their specific or combined utility as diagnostic and prognostic biomarkers. NfL, CHIT1 and miR-181b all showed significantly higher levels in ALS subjects compared to controls, with NfL showing the most effective diagnostic performance. Importantly, all three biomarkers were increased compared to neurodegenerative disease controls and, specifically, to patients with Alzheimer’s disease (AD; N = 44), with NfL and CHIT1 being also higher in ALS than in alpha-synucleinopathies (N = 22). Notably, ALS patients displayed increased CHIT1 levels despite having, compared to controls, a higher prevalence of a polymorphism lowering CHIT1 expression. While no relationship was found between CSF miR-181b and clinical measures in ALS (disease duration, functional disability, and disease progression rate), CSF NfL was the best independent predictor of disease progression and survival. This study deepens our knowledge of ALS biomarkers, highlighting the relative specificity of CHIT1 for ALS among neurodegenerative diseases and appraising the potential diagnostic utility of CSF miR-181b.

Hyaluronan in lung, in plasma as pathogenic and prediction factor of acute respiratory distress syndrome: A systematic review

This investigation aims to study contemporary literature pertaining to the involvement of hyaluronate in the pathogenesis of diverse medical conditions, encompassing coronavirus-induced pulmonary injury, while also exploring its potential utility as a prognostic indicator for assessing the severity of COVID-19. This study conducted a comprehensive examination of hyaluronic acid’s multifaceted role in physiological processes and disease, with a specific focus on its implications in COVID-induced lung damage. The research provided an in-depth analysis of the intricate mechanisms and fundamental patterns governing these biological phenomena, elucidating essential interactions and pathways. Of particular significance in this investigation was the potential diagnostic utility of hyaluronic acid in assessing the severity of acute respiratory distress syndrome (ARDS), including COVID-19. Through a rigorous examination of hyaluronic acid concentration levels, researchers sought to assess its potential as an early prognostic indicator, thereby providing valuable insights for clinical diagnostics. Furthermore, the study explored the therapeutic prospects related to hyaluronic acid, emphasizing its involvement in various pathological processes. It suggested that targeting hyaluronic acid could represent a promising avenue for drug development, potentially leading to the creation of innovative pharmaceutical agents

Abstract B023: Unravelling fibroinflammatory and immune signatures for pancreatic cancer early detection

Abstract Pancreatic cancer is associated with late-stage diagnosis and poor patient prognosis. The disease is characterised by chronic dysregulated immunity and inflammation. Moreover, chronic pancreatitis is linked with an increased risk of pancreatic cancer. Tissue remodelling during disease progression involves a complex network of cells and a crosstalk between cancer cells, stromal components and immune cells, but it remains poorly explored. Thus, to improve the outcome of patients, there is a need for better understanding of biological changes associated with the initiation and progression of pancreatic cancer at the molecular and spatial levels. Using samples obtained from our Accelerated Diagnosis of neuroEndocrine and Pancreatic TumourS (ADEPTS) biobank, we have evaluated blood samples for the discovery of non- invasive biomarkers to discriminate patients with pancreatic cancer from symptomatic patients without cancer. In this pilot study, the transcriptome of peripheral blood mononuclear cells (PBMCs), isolated using a ficoll gradient was explored to identify PDAC-associated mRNA signatures. Moreover, the Olink® Explore Inflammation and Oncology panels were performed in a cohort of 40 patients with early stage (I-II) PDAC, 40 chronic pancreatitis samples and 40 control patients with benign symptomatic upper GI diseases. To validate and enrich these results, a panel of inflammatory and immune markers including immunoglobulins, cytokines and chemokines was also run using multiplex techniques. Symptoms observed in our cohort were also analysed together with clinical (e.g diabetic status, biochemistry profile) and demographic data (age, sex, ethnicity, post code etc). We have identified mRNA transcriptome signatures and differentially expressed proteins associated with inflammatory and immunological processes in PDAC compared to benign symptomatic samples. Our results offer potential diagnostic utility for early-stage disease. Additionally, our unique cohort of patient samples comprising of those with confirmed PDAC and those presenting to clinic with benign disease and similar non-specific symptoms, allows for the development of non-invasive diagnostic tests with better translational applicability. Future work will involve the validation of these findings in a larger patient cohort and the use of NanoString GeoMx® Digital Spatial Profiler to perform whole transcriptome and proteome profiling in human chronic pancreatic and pancreatic cancer tissue samples. Citation Format: Kyra Fraser, Maria Rosado, Stephen P Pereira, Pilar Acedo. Unravelling fibroinflammatory and immune signatures for pancreatic cancer early detection [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr B023.

Synthesis and screening of a library of Lewisx deoxyfluoro-analogues reveals differential recognition by glycan-binding partners

Glycan-mediated interactions play a crucial role in biology and medicine, influencing signalling, immune responses, and disease pathogenesis. However, the use of glycans in biosensing and diagnostics is limited by cross-reactivity, as certain glycan motifs can be recognised by multiple biologically distinct protein receptors. To address this specificity challenge, we report the enzymatic synthesis of a 150-member library of site-specifically fluorinated Lewisx analogues (‘glycofluoroforms’) using naturally occurring enzymes and fluorinated monosaccharides. Subsequent incorporation of a subset of these glycans into nanoparticles or a microarray revealed a striking spectrum of distinct binding intensities across different proteins that recognise Lewisx. Notably, we show that for two proteins with unique binding sites for Lewisx, glycofluoroforms exhibited enhanced binding to one protein, whilst reduced binding to the other, with selectivity governed by fluorination patterns. We finally showcase the potential diagnostic utility of this approach in glycofluoroform-mediated bacterial toxin detection by lateral flow.

Exosome-mediated transfer of lncRNA RP3-340B19.3 promotes the progression of breast cancer by sponging miR-4510/MORC4 axis

BackgroundThis study aims to explore the molecular mechanism of lncRNA RP3-340B19.3 on breast cancer cell proliferation and metastasis and clinical significance of lncRNA RP3-340B19.3 for breast cancer.MethodsThe subcellular localization of lncRNA RP3-340B19.3 was identified using RNA fluorescence in situ hybridization (FISH). The expression of lncRNA RP3-340B19.3 in breast cancer cells, breast cancer tissues, as well as the serum and serum exosomes of breast cancer patients, was measured through quantitative RT-PCR. In the in vitro setting, we conducted experiments to observe the effects of RP3-340B19.3 on both cell migration and proliferation. This was achieved through the utilization of transwell migration assays as well as clone formation assays. Meanwhile, transwell migration assays and clone formation assays were used to observe the effects of MDA-MB-231-exosomes enriched in RP3-340B19.3 on breast cancer microenvironment cells MCF7 and BMMSCs. Additionally, western blotting techniques were used to assess the expression levels of proteins associated with essential cellular processes such as proliferation, apoptosis, and metastasis. In vivo, the impact of RP3-340B19.3 knockdown on tumour weight and volume was observed within a nude mice model. We aimed to delve into the intricate molecular mechanisms involving RP3-340B19.3 by using bioinformatics analysis, dual luciferase reporter gene experiments and western blotting. Moreover, the potential correlations between RP3-340B19.3 expression and various clinical pathological characteristics were analyzed.ResultsOur investigation revealed that RP3-340B19.3 was expressed in both the cytoplasm and nucleus, with a noteworthy increase in breast cancer cells. Notably, we found that RP3-340B19.3 exerted a promoting influence on the proliferation and migration of breast cancer cells, both in vitro and in vivo. MDA-MB-231-exosomes enriched in RP3-340B19.3 promoted the proliferation and migration of MCF7 and BMMSCs in vitro. Mechanistically, RP3-340B19.3 demonstrated the capability to modulate the expression of MORC4 by forming a complex with miR-4510. This interaction subsequently triggered the activation of the NF-κB and Wnt-β-catenin signaling pathways. Furthermore, our study highlighted the potential diagnostic utility of RP3-340B19.3. We discovered its presence in the serum and exosomes of breast cancer patients, showing promising efficacy as a diagnostic marker. Notably, the diagnostic potential of RP3-340B19.3 was particularly significant in relation to distinguishing between different pathological types of breast cancer and correlating with tumour diameter.ConclusionOur findings establish that RP3-340B19.3 plays a pivotal role in driving the proliferation and metastasis of breast cancer. Additionally, exosomes enriched in RP3-340B19.3 could influence MCF7 and BMMSCs in tumour microenvironment, promoting the progression of breast cancer. This discovery positions RP3-340B19.3 as a prospective novel candidate for a tumour marker, offering substantial potential in the realms of breast cancer diagnosis and treatment strategies.

The Potential Diagnostic Utility of SMAD4 and ACCS in the Context of Inflammation in Type 2 Diabetes Mellitus Patients.

The search for new parameters for the prediction of type 2 diabetes mellitus (T2DM) or its harmful consequences remains an important field of study. Depending on the low-grade inflammatory nature of diabetes, we investigated three proteins in T2DM patients: 1-aminocyclopropane-1-carboxylate synthase (ACCS), granulocyte-colony-stimulating factor (G-CSF), and Sma Mothers Against Decapentaplegic homolog-4 (SMAD4). In brief, sixty T2DM and thirty healthy controls had their serum levels of ACCS, G-CSF, SMAD4, and insulin tested using the ELISA method. The insulin resistance (IR) parameter (HOMA2IR), beta-cell function percentage (HOMA2%B), and insulin sensitivity (HOMA2%S) were all determined by the Homeostasis Model Assessment-2 (HOMA2) calculator. The predictability of these protein levels was investigated by neural network (NN) analysis and was associated with measures of IR. Based on the results, ACCS, G-CSF, and SMAD4 increased significantly in the T2DM group compared with the controls. Their levels depend on IR status and inflammation. The multivariate GLM indicated the independence of the levels of these proteins on the covariates or drugs taken. The receiver operating characteristic area under the curve (AUC) for the prediction of T2DM using NN analysis is 0.902, with a sensitivity of 71.4% and a specificity of 93.8%. The network predicts T2DM well with predicted pseudoprobabilities over 0.5. The model's predictive capability (normalized importance) revealed that ACCS is the best model (100%) for the prediction of T2DM, followed by G-CSF (75.5%) and SMAD4 (69.6%). It can be concluded that ACCS, G-CSF, and SMAD4 are important proteins in T2DM prediction, and their increase is associated with the presence of inflammation.