Platelet Markers Research Articles

Endothelial dysfunction markers syndecan-1 and thrombomodulin are associated with higher albuminuria levels in type 2 diabetes with no history of clinical cardiovascular disease

IntroductionIndividuals with type 2 diabetes and increased albuminuria, a well-established marker of microvascular complications, are at a higher risk for cardiovascular disease (CVD) and premature mortality. Therefore, a better understanding of the underlying pathophysiology is needed to improve risk stratification and tailor prevention and intervention. MethodsWe conducted a cross-sectional study including 463 individuals with type 2 diabetes, various degrees of albuminuria and without CVD. We analysed the association between albuminuria and markers of endothelial function (thrombomodulin and syndecan-1), thrombin generation (thrombin-antithrombin complex, prothrombin fragment 1 + 2), fibrinogen, platelet function (activation using soluble plasma selectin and aggregation using Multiplate® Analyzer) using regression models. ResultsIn the study cohort 33 % were women, the mean ± SD age was 65 ± 9 years, and median [IQR] diabetes duration was 15 [9–20] years. In total, 344 (74 %) individuals had normal albuminuria, 87 (19 %) moderately- and 32 (7 %) severely increased albuminuria levels. Higher markers of endothelial function and fibrinogen were independently associated with higher albuminuria levels (p < 0.01). No association between albuminuria and markers of thrombin generation and platelet was demonstrated. ConclusionWe demonstrated an independent association between albuminuria and markers of endothelial function and fibrinogen in individuals with type 2 diabetes and no history of CVD.

Choice of blood collection methods influences extracellular vesicles counts and miRNA profiling.

Circulating RNAs have been investigated systematically for over 20years, both as constituents of circulating extracellular vesicles (EVs) or, more recently, non-EV RNA carriers, such as exomeres and supermeres. The high level of variability and low reproducibility rate of EV/extracellular RNA (exRNA) results generated even on the same biofluids promoted several efforts to limit pre-analytical variability by standardizing sample collection and sample preparation, along with instrument validation, setup and calibration. Anticoagulants (ACs) are often chosen based on the initial goal of the study and not necessarily for the later EV and/or exRNA analyses. We show the effects of blood collection on EV size, abundance, and antigenic composition, as well on the miRNAs. Our focus of this work was on the effect of ACs on the number and antigenic composition of circulating EVs and on a set of circulating miRNA species, which were shown to be relevant as disease markers in several cancers and Alzheimer's disease. Results show that while the number of plasma EVs, their relative size, and post-fluorescence labeling profile varied with each AC, their overall antigenic composition, with few exceptions, did not change significantly. However, the number of EVs expressing platelet and platelet-activation markers increased in serum samples. For overall miRNA expression levels, EDTA was a better AC, although this may have been associated with stimulation of cells in the blood collection tube. Citrate and serum rendered better results for a set of miRNAs that were described as circulating markers for Alzheimer's disease, colon, and papillary thyroid cancers.

Adherence to the Healthy Nordic Food Index is associated with reduced plasma levels of inflammatory markers in patients with heterozygous familial hypercholesterolemia

Background and aimsFamilial hypercholesterolemia (FH) is an inherited disease associated with hypercholesterolemia, and dietary treatment is part of the treatment. We aimed to assess the dietary pattern in relation to the Healthy Nordic Food Index (HNFI) in adults with and without heterozygous FH (HeFH), and to examine the associations between dietary quality and biomarkers related to cardiovascular disease in adults with HeFH. MethodsWe included 205 adults (≥18 years) with HeFH who received follow-up at the Lipid Clinic in Oslo and compared them to controls (n=228). Dietary intake was assessed using a food frequency questionnaire and dietary quality was assessed using the HNFI. Blood samples were analysed for levels of blood lipids, plasma fatty acids (FAs), and markers of inflammation and platelet activation. ResultsThe HeFH patients (median 60 years; 50.2% female; 25.9% in secondary prevention) had lower intake of total and saturated fat compared to controls (32.6 energy percent (E%) vs. 34.9E%, and 9.6 E% vs 12.0 E%, respectively; p<0.001 for both). In the HeFH patients, increasing dietary quality was associated with increased plasma levels of the n-3 polyunsaturated FAs (PUFAs) eicosapentaenoic acid and docosahexaenoic acid, and the n-6 PUFA linoleic acid, and lower plasma levels of the inflammatory cytokines Tumor Necrosis Factor and interleukin-6, and of the platelet-derived inflammatory cytokines Platelet Factor 4 and Neutrophil-Activating Peptide-2. ConclusionNorwegian patients with HeFH followed up at a Lipid Clinic eat healthier than controls. Adherence to a healthy dietary pattern is associated with higher plasma levels of n-3 and n-6 PUFA, and lower levels of inflammatory markers, including platelet markers. This may suggest that adherence to an overall healthy dietary pattern might be beneficial for HeFH patients independent of the cholesterol-lowering effect of the diet.

Comprehensive Analysis of a Platelet- and Coagulation-Related Prognostic Gene Signature Identifies CYP19A1 as a Key Tumorigenic Driver of Colorectal Cancer.

The intricate interplay between the platelet-coagulation system and the progression of malignant tumors has profound therapeutic implications. However, a thorough examination of platelet and coagulation markers specific to colorectal cancer (CRC) is conspicuously absent in the current literature. Consequently, there is an urgent need for further exploration into the mechanistic underpinnings of these markers and their potential clinical applications. By integrating RNA-seq data and clinicopathological information from patients with CRC in the cancer genome atlas, we identified genes related to the platelet-coagulation system using weighted gene co-expression networks and univariate Cox analysis. We established a prognostic risk model based on platelet- and coagulation-related genes using Lasso Cox regression analysis and validated the model in two independent CRC cohorts. We explored potential biological functional disparities between high-risk and low-risk groups through comprehensive bioinformatics analysis. Our findings indicate that colorectal cancer patients classified as high-risk generally exhibit poorer prognoses. Moreover, the model's risk scores were associated with the differential composition of the immune tumor microenvironment, suggesting its applicability to infer immunotherapy responsiveness. Cellular functional experiments and animal experiments indicated that CYP19A1 expression in CRC influences malignant phenotype and platelet activation. In summary, we present a novel platelet- and coagulation-related risk model for prognostic assessment of patients with CRC and confirm the important role of CYP19A1 in promoting malignant progression of CRC.

Double-stranded DNA enhances platelet activation, thrombosis, and myocardial injury via cyclic GMP-AMP synthase

Abstract Aims Elevated dsDNA levels in STEMI patients are associated with increased infarct size and worse clinical outcomes. However, the direct effect of dsDNA on platelet activation remains unclear. This study aims to investigate the direct influence of dsDNA on platelet activation, thrombosis, and the underlying mechanisms. Methods and Results Analysis of clinical samples revealed elevated plasma dsDNA levels in STEMI patients, which positively correlated with platelet aggregation and markers of neutrophil extracellular traps (NETs) such as MPO-DNA and CitH3. Platelet assays demonstrated the activation of the cGAS-STING pathway in platelets from STEMI patients. DsDNA directly potentiated platelet activation and thrombus formation. Mechanistic studies using G150 (cGAS inhibitor), H151 (STING inhibitor), and MCC950 (NLRP3 inhibitor), as well as cGAS-/-, STING-/- and NLRP3-/- mice showed that dsDNA activated cGAS, a previously unreported DNA sensor in platelets, and induced activation of the STING/NLRP3/caspase-1/IL-1β axis. This cascade enhanced platelet activation and thrombus formation. Platelet cGAS depletion or Palbociclib, a cGAS-STING inhibitor, approved by the FDA for advanced breast cancer, ameliorated myocardial ischemia-reperfusion injury in ApoE-/- mice fed with a high-fat diet for 12 weeks. Conclusions These results suggested that dsDNA is a novel driver of platelet activation and thrombus formation in STEMI patients. Translational Perspective ST-elevated myocardial infarction (STEMI) patients exhibit high levels of plasma double-stranded DNA (dsDNA), which directly potentiates platelet activation through the platelet cGAS/STING/NLRP3/caspase-1/IL-1β signaling pathway. STEMI patients may benefit from cGAS inhibition in the prevention of platelet hyperactivity and thrombus formation.

Platelet transfusion enhances pro-aggregatory status shortly aftercoronary artery bypass grafting (CABG while modulating platelet pro-inflammatory state 1-week post-surgery.

During coronary artery bypass grafting (CABG), the surgical procedure, particularly the manipulation of the major arteries of the heart, induces a significant inflammatory state that may compromise platelet function to the extent that platelet transfusion is required. Given stored platelets as a major source of biological mediators, this study investigates the effects of platelet transfusion on the major pro-aggregatory, pro-inflammatory and immunomodulatory markers of platelets. Platelets from 20 patients, 10 who received platelet transfusion and 10 without, were subjected to flow cytometery where P-selectin and CD40 ligand (CD40L) expressions and PAC-1 binding (activation-specific anti GPIIb/GPIIIa antibody)analysed at five-time points of 24 h before surgery, immediately, 2 h, 24 h and 1 week after surgery. Analysis of intra-platelet transforming growth factor-beta-1 (TGF-β1) was also conducted using western blotting. Patients with platelet transfusion showed increased levels of P-selectin, CD40L and intra-platelet TGF-β1 2-h after surgery compared to those without transfusion (p < 0.05). PAC-1 binding was increased 24 h after surgery in transfused patients (p < 0.05). Given the significant post-transfusion elevation of platelet TGF-β1, P-sel/CD40L reduction in transfused patients a week after was of much interest. This study showed for the first time the significant effects of platelet transfusion on the pro-inflammatory, pro-aggeregatory and immunomodulatory state of platelets in CABG patients, which manifested with immediate, midterm and delayed consequences. While the increased pro-inflammatory conditions manifested as an immediate effect of platelet transfusion, the pro-aggregatory circumstances emerged 24h post-transfusion. A week after surgery, attenuations of pro-inflammatory markers of platelets in transfused patients were shown, which might be due to the immunomodulatory effects of TGF-β1.

Engineering Platelet Membrane Imitating Nanoparticles for Targeted Therapeutic Delivery.

Platelet Membrane Imitating Nanoparticles (PMINs) is a novel drug delivery system that imitates the structure and functionality of platelet membranes. PMINs imitate surface markers of platelets to target specific cells and transport therapeutic cargo. PMINs are engineered by incorporating the drug into the platelet membrane and encapsulating it in a nanoparticle scaffold. This allows PMINs to circulate in the bloodstream and bind to target cells with high specificity, reducing off-target effects and improving therapeutic efficacy. The engineering of PMINs entails several stages, including the separation and purification of platelet membranes, the integration of therapeutic cargo into the membrane, and the encapsulation of the membrane in a nanoparticle scaffold. In addition to being involved in a few pathological conditions including cancer, atherosclerosis, and rheumatoid arthritis, platelets are crucial to the body's physiological processes. This study includes the preparation and characterization of platelet membrane-like nanoparticles and focuses on their most recent advancements in targeted therapy for conditions, including cancer, immunological disorders, atherosclerosis, phototherapy, etc. PMINs are a potential drug delivery system that combines the advantages of platelet membranes with nanoparticles. The capacity to create PMMNs with particular therapeutic cargo and surface markers provides new possibilities for targeted medication administration and might completely change the way that medicine is practiced. Despite the need for more studies to optimize the engineering process and evaluate the effectiveness and safety of PMINs in clinical trials, this technology has a lot of potential.

Deciphering Abnormal Platelet Subpopulations in COVID-19, Sepsis and Systemic Lupus Erythematosus through Machine Learning and Single-Cell Transcriptomics.

This study focuses on understanding the transcriptional heterogeneity of activated platelets and its impact on diseases such as sepsis, COVID-19, and systemic lupus erythematosus (SLE). Recognizing the limited knowledge in this area, our research aims to dissect the complex transcriptional profiles of activated platelets to aid in developing targeted therapies for abnormal and pathogenic platelet subtypes. We analyzed single-cell transcriptional profiles from 47,977 platelets derived from 413 samples of patients with these diseases, utilizing Deep Neural Network (DNN) and eXtreme Gradient Boosting (XGB) to distinguish transcriptomic signatures predictive of fatal or survival outcomes. Our approach included source data annotations and platelet markers, along with SingleR and Seurat for comprehensive profiling. Additionally, we employed Uniform Manifold Approximation and Projection (UMAP) for effective dimensionality reduction and visualization, aiding in the identification of various platelet subtypes and their relation to disease severity and patient outcomes. Our results highlighted distinct platelet subpopulations that correlate with disease severity, revealing that changes in platelet transcription patterns can intensify endotheliopathy, increasing the risk of coagulation in fatal cases. Moreover, these changes may impact lymphocyte function, indicating a more extensive role for platelets in inflammatory and immune responses. This study identifies crucial biomarkers of platelet heterogeneity in serious health conditions, paving the way for innovative therapeutic approaches targeting platelet activation, which could improve patient outcomes in diseases characterized by altered platelet function.

Correlation analysis of peripheral platelet markers and disease phenotypes in Alzheimer's disease.

Platelets serve as the primary peripheral reservoir of amyloid beta (Aβ). However, there is limited research on platelet markers in routine blood examinations, particularly with regard to the large platelet ratio (P-LCR) in Alzheimer's disease (AD). This study included 512 AD patients and 205 healthy controls (HCs). Platelet markers and apolipoprotein E (APOE) 4 status were assessed in all participants. The study revealed that P-LCR was significantly elevated in AD patients compared to HCs. In AD patients carrying APOE4, P-LCR significantly negatively correlated with Montreal Cognitive Assessment scores. There was an observed increasing trend in the rate of change in P-LCR with disease progression. Binary logistic regression analysis indicated that P-LCR may constitute a risk factor for AD, after adjusting for age, sex, APOE4, and body mass index. P-LCR is associated with disease severity in AD patients carrying APOE4. P-LCR may be a promising marker to reflect platelet activity in AD patients. P-LCR significantly negatively correlated with MoCA scores in AD patients with APOE4. The rate of change in P-LCR showed an increasing trend with disease progression. P-LCR may be a risk factor for AD.

Removal of endothelial surface-associated von villebrand factor suppresses accelerate datherosclerosis after myocardial infarction

BackgroundThromboinflammation involving platelet adhesion to endothelial surface-associated von Willebrand factor (VWF) has been implicated in the accelerated progression of non-culprit plaques after MI. The aim of this study was to use arterial endothelial molecular imaging to mechanistically evaluate endothelial-associated VWF as a therapeutic target for reducing remote plaque activation after myocardial infarction (MI).MethodsHyperlipidemic mice deficient for the low-density lipoprotein receptor and Apobec-1 underwent closed-chest MI and were treated chronically with either: (i) recombinant ADAMTS13 which is responsible for proteolytic removal of VWF from the endothelial surface, (ii) N-acetylcysteine (NAC) which removes VWF by disulfide bond reduction, (iii) function-blocking anti-factor XI (FXI) antibody, or (iv) no therapy. Non-ischemic controls were also studied. At day 3 and 21, ultrasound molecular imaging was performed with probes targeted to endothelial-associated VWF A1-domain, platelet GPIbα, P-selectin and vascular cell adhesion molecule-1 (VCAM-1) at lesion-prone sites of the aorta. Histology was performed at day 21.ResultsAortic signal for P-selectin, VCAM-1, VWF, and platelet-GPIbα were all increased several-fold (p < 0.01) in post-MI mice versus sham-treated animals at day 3 and 21. Treatment with NAC and ADAMTS13 significantly attenuated the post-MI increase for all four molecular targets by > 50% (p < 0.05 vs. non-treated at day 3 and 21). On aortic root histology, mice undergoing MI versus controls had 2–4 fold greater plaque size and macrophage content (p < 0.05), approximately 20-fold greater platelet adhesion (p < 0.05), and increased staining for markers of platelet transforming growth factor-β1 signaling. Accelerated plaque growth and inflammatory activation was almost entirely prevented by ADAMTS13 and NAC. Inhibition of FXI had no significant effect on molecular imaging signal or plaque morphology.ConclusionsPlaque inflammatory activation in remote arteries after MI is strongly influenced by VWF-mediated platelet adhesion to the endothelium. These findings support investigation into new secondary preventive therapies for reducing non-culprit artery events after MI.

Targeting purinergic signaling in platelets ameliorates the bacterial burden and reduces the cytokine storm in the peritoneal cavity during intra-abdominal sepsis

Sepsis is a complex clinical syndrome of the host response to an infection, involving a pathogenic cytokine storm that drives inflammatory injury in multiple organ systems. The abdomen is a major site of sepsis, contributing to high mortality. Platelets express the purinergic receptors P2Y1 and P2Y12; once activated, these receptors mediate the secretion of pro-inflammatory mediators that contribute to organ damage in sepsis. Systemic blockade of P2Y1 or P2Y12 decreased organ damage and inflammatory cytokine levels in a sex-related manner during sepsis. Here, we hypothesize that blocking P2Y1 or P2Y12 activity enhances pathogen clearance in the peritoneal cavity during intra-abdominal sepsis in a sex-specific manner. Male and female C57BL6/6J mice underwent sham surgery or cecal ligation and puncture (CLP) to induce sepsis. P2Y1 or P2Y12 antagonists, MRS2279 (10 mg/kg) and Ticagrelor (30 mg/kg) respectively, were administered intraperitoneally after surgery. The peritoneal cavity fluid (PCF) was collected 24 hours post-surgery by peritoneal lavage. To assess cell recruitment in the cavity, we measured leukocyte and platelet counts using Hemavet® Multispecies Hematology System. To assess inflammation, we measured PCF levels of the cytokines MIP-1α and Rantes as well as bacterial clearance. The ileum of the small intestinal tissue samples were collected 24 hours post-surgery and platelet infiltration was measured by platelet factor 4 (PF4) content in the tissue using an ELISA kit. In response to the P2Y1 or P2Y12 blocker, we noted a significant reduction in the bacteria content in the PCF of CLP mice. Blocking P2Y12 but not P2Y1 reduced leukocyte recruitment, while platelet counts were reduced only with P2Y1 blockade in septic males. The sepsis-induced increase in MIP-1α levels was not noted upon P2Y1 but not P2Y12 blockage in the PCF of septic males. However, Rantes levels in the PCF were reduced for either treatment in males compared to the untreated CLP. The platelet marker PF4 in the ileum was increased during sepsis but reduced after either treatment in sepsis males as compared to the untreated CLP. In female CLP mice, a significant reduction in bacterial burden in the PCF in response to the P2Y1 but not the P2Y12 blocker was shown. Blocking P2Y1 or P2Y12 reduced leukocyte recruitment and platelet counts in female CLP mice. As observed in males, PF4 was increased in the ileum during sepsis but reduced following exposure to either treatment in females. No change was noted for MIP-1α or Rantes levels in the females among all the septic groups. Our data indicate that blocking purinergic signaling and in particular P2Y12 in the peritoneal cavity in males prevents bacterial spreading and diminishes cytokine levels, indicating that it could be an effective treatment for intra-abdominal sepsis. However, in female mice blocking the receptor P2Y1, but not P2Y12 has shown to be optimal. Targeting purinergic signaling is a promising sex-specific therapeutic strategy for sepsis. This work was supported by the Research grant AI156627-01 (EL) from the National Institute of Health. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Immature platelets and platelet reactivity in patients with acute ST-segment Elevation Myocardial Infarction using whole blood flow cytometry with SYTO-13 staining

BackgroundReduced effect of antiplatelet therapy has been reported in patients with ST-segment elevation myocardial infarction (STEMI). Multiple factors may concur to explain this, including increased amount of highly reactive immature platelets. ObjectivesTo investigate the association between immature platelets and reactivity determined with multicolour flow cytometry using the SYTO-13 dye in STEMI patients. MethodsWe conducted an observational study of 59 patients with acute STEMI. Blood samples were obtained within 24 h after admission and after loading doses of dual antiplatelet therapy. For comparison, samples were obtained from 50 healthy individuals. Immature platelets and platelet reactivity were investigated using multicolour flow cytometry including the SYTO-13 dye that binds to platelet RNA and thus provides a method for subdividing platelets into immature and mature platelets. Additionally, we assessed platelet aggregation, serum-thromboxane B2 levels and standard immature platelet markers. ResultsImmature platelets were more reactive than mature platelets in both STEMI patients and healthy individuals (p-values < 0.05). STEMI patients had lower platelet aggregation and thromboxane B2 levels than healthy individuals. We found a positive association between automatically determined immature platelet markers and CD63 expression on activated platelets (Spearman's rho: 0.27 to 0.58, p-values < 0.05). ConclusionsOur study shows that immature platelets identified with a multicolour flow cytometric method using the SYTO-13 dye are more reactive than mature platelets in patients with acute STEMI and in healthy individuals. The presence of immature platelets may be important for the overall platelet reactivity, which may have implications for the effect of antiplatelet therapy.

Effectiveness Of Platelet Markers In Estimating The Amount Of Intraoperative Bleeding In Vertebra Surgery

AIMS: One of the most important steps in perioperative bleeding management is the predetermination of the patient at risk. Even if there is no hemostatic abnormality in vertebral surgery, which is a major surgery, severe bleeding can be encountered and many perioperative blood transfusions might be required. To date, there are no studies in the literature examining platelet markers in terms of predictivity in intraoperative bleeding during vertebral surgeries. In this study; we investigated the effectiveness of preoperative mean platelet volume (MPV), platelet distribution width (PDW), plateletcrit (PCT), mean platelet volume lymphocyte count ratio (MPVLR), and platelet count to lymphocyte count ratio (PLR) values in predicting the amount of intraoperative bleeding in patients who will undergo vertebral surgery. &#x0D; METHODS: After the approval from the ethics committee, patients who were planned for vertebral surgery by the neurosurgeon were identified. Patient demographic data such as gender, age, ASA score and body mass index (BMI), MPV, PDW, PCT, MPVLR, and PLR values were recorded. Total surgery time, the number of vertebral transpedicular screwing, and laminectomy levels performed as surgical procedures were recorded. The amount of intraoperative bleeding was determined by calculating the amount of blood accumulated in the aspirator and the amount of blood collected by the sponge during surgery. &#x0D; RESULTS: Of the 63 patients included in the study, 60.3% (n=38) were female and 39.7% (n=25) were male. The mean age of the patients was 54.7 ± 11 years and the mean BMI was 29.6 ± 4.6. No correlation was found between the amount of bleeding and gender, age, MPV, PDW, PCT, MPVLR, or PLR values. A significantly high correlation was found between amount of bleeding and BMI, duration of surgery, and the number of transpedicular screwing segments. &#x0D; CONCLUSION: Platelet markers do not appear to have an effect on the amount of bleeding. However, as the BMI increases, the duration of surgery prolongs and the number of transpedicular screw segments increases for which the amount of bleeding increases.

Profile of matrix‐entrapped extracellular vesicles of microenvironmental and infiltrating cell origin in decellularized colorectal cancer and adjacent mucosa

AbstractCellular elements that infiltrate and surround tumours and pre‐metastatic tissues have a prominent role in tumour invasion and growth. The extracellular vesicles specifically entrapped and stored within the extracellular matrix (ECM‐EVs) may reflect the different populations of the tumour microenvironment and their change during tumour progression. However, their profile is at present unknown. To elucidate this aspect, we isolated and characterized EVs from decellularized surgical specimens of colorectal cancer and adjacent colon mucosa and analyzed their surface marker profile. ECM‐EVs in tumours and surrounding mucosa mainly expressed markers of lymphocytes, natural killer cells, antigen‐presenting cells, and platelets, as well as epithelial cells, representing a multicellular microenvironment. No difference in surface marker expression was observed between tumour and mucosa ECM‐EVs in stage II‐III tumours. At variance, in the colon mucosa adjacent to stage IV carcinomas, ECM‐EV profile showed a significantly increased level of immune, epithelial and platelet markers in comparison to the matrix of the corresponding tumour. The increase of EVs from immune cells and platelets was not observed in the mucosa adjacent to low‐stage tumours. In addition, CD25, a T‐lymphocyte marker, resulted specifically overexpressed by ECM‐EVs from stage IV carcinomas, possibly correlated with the pro‐tolerogenic environment found in the corresponding tumour tissue. These results outline the tissue microenvironmental profile of EVs in colorectal carcinoma‐derived ECM and unveil a profound change in the healthy mucosa adjacent to high‐stage tumours.

Evaluation of hepatic toxicity in autoimmune hemolytic anemia (AIHA) and Evans syndrome patients: a single-center Egyptian study

IntroductionBenign auto-immune illnesses include Evans syndrome (ES) and auto-immune hemolytic anemia (AIHA). Despite being benign in nature, the patients’ livers are burdened by the disease’s chronicity and the accompanying problems beyond the course of treatment. An additional burden stems from HCV infection, of which a significant proportion of Egyptians are positive. The purpose of this study was to identify the hepatotoxicity risks and the variables that influence the prognosis and survival of patients with AIHA/ES. There are 126 AIHA patients in this observational study, which is retrospective. From June 2009 to March 2021, patients visited the Haematology Unit of the Oncology Centre in Egypt. One hundred and sixteen patients have available data.ResultsThere was no significant difference between primary and secondary AIHA groups as regards baseline hemoglobin (Hb), bilirubin, LDH, or reticulocyte count. Thirty-four patients (29.31%) had HCV-positive tests and 1 patient (0.9%) had HBV. There was no difference between HCV-positive and negative cases as regards mean Hb concentration, mean platelet, or immune markers (P > 0.05). AIHA patients with HCV-positive showed a significantly higher relapse rate (56%) than HCV-negative patients (32%) (P = 0.034). HCV positivity and low platelet counts at diagnosis were poor predictors for overall survival (OS) (P 0.022 and 0.04, respectively). Median OS was significantly better in patients with no viral hepatitis infection (1101 days, 95% CI 592–2068) than in patients with positive HCV infection (521, 95% CI 326–1325) (P = 0.019).ConclusionsAzathioprine is the least hepatotoxic in AIHA patients under treatment. Viral hepatitis represents a superadded damage to the liver besides AIHA concerning clinical characteristics and outcomes.

Assessment of Platelet Indices as Biomarker for Neonatal Sepsis

Introduction: There is a need to increase the diagnostic capability of the traditional septic screen by adding additional markers. Platelet indices are one such marker that is readily available in routine hemogram reports. Materials and Methods: In this cross-sectional study, all newborn babies were recruited who had signs or symptoms of sepsis or had high-risk factors. Those who had clinical sepsis with positive cultures and/or a positive sepsis screen were classified under group “cases” ( n = 174), whereas all neonates suspected to have sepsis but who had a negative blood culture and a negative sepsis screen were classified under group “control” ( n = 126). Blood culture, sepsis screen, and platelet indices [platelet count, mean platelet volume (MPV), and platelet distribution width (PDW)] were performed on all babies. Babies with congenital or acquired causes of platelet abnormalities (including medication use by mothers with antiplatelet activity) were excluded from this study. Results: In this study, a total of 300 newborn babies were recruited for final analyses. Overall culture positivity was 38%. The mean platelet count was lower in the cases group ( P &lt; 0.001). MPV and PDW were higher in septic babies with statistically significant differences ( P &lt; 0.0001 and P = 0.012, respectively). Thrombocytopenia was the most sensitive marker (85.60%). However, it had low specificity (28.2%). On combining all three platelet markers, specificity increased to 48.4% (with a sensitivity of 64.5%) in detecting babies with neonatal sepsis. The sensitivity of the septic screen alone was 58% (with a specificity of 32.62%). When combining the sepsis screen and platelet indices, the specificity for the diagnosis of neonatal sepsis increased to 62.6%. On receiver operator curve analyses, thrombocytopenia had the highest area under the curve (0.692), followed by MPV (0.644). Conclusion: Platelet indices may be used as sensitive markers in combination with traditional sepsis screening to make an early diagnosis of neonatal sepsis.

Platelet Proteome Reveals Novel Targets for Hypercoagulation in Pseudoexfoliation Syndrome.

Pseudoexfoliation syndrome (PEX) is characterized by the accumulation of abnormal extracellular matrix material in ocular and non-ocular tissues, including blood vessel walls. Clot-forming dysfunction might be responsible for venous thrombosis in PEX. We investigated global coagulation, the proteome, and functions of platelets in PEX patients and aimed to determine prognostic biomarkers for thrombosis risk in PEX. Peripheral blood was collected from PEX and retinal vein occlusion (RVO) patients, and age-sex matched controls. Viscoelastic hemostasis was evaluated by rotational thromboelastometry (ROTEM). Platelet markers (CD41, CD42, CD61, and CD62p) and endothelial markers (P-selectin, E-selectin, and von Willebrand factor) were investigated by flow cytometry and ELISA, respectively. The platelet proteome was analyzed by 2D fluorescence difference gel electrophoresis followed by mass spectrometry. Clot formation time (CFT) is significantly reduced in PEX patients compared to the controls (p < 0.05). P-selectin levels were higher in PEX patients than in controls (p < 0.05); E-selectin and von Willebrand factor remained unchanged. The monitorization of CFT by ROTEM, and soluble P-selectin, may help assess thrombotic risk in PEX patients. Proteomic analysis revealed differential expression of Profilin-1 in platelets. Profilin-1 regulates the stability of actin-cytoskeleton and may contribute to impaired platelet hemostatic functions. Increased P-selectin levels together with impaired coagulation dynamics might be responsible for the thrombotic events in PEX disease.

Tetraspanin profiles of serum extracellular vesicles reflect functional limitations and pain perception in knee osteoarthritis.

Emerging evidence suggests that extracellular vesicles (EVs) can play roles in inflammatory processes and joint degradation in primary osteoarthritis (OA), a common age-associated joint disease. EV subpopulations express tetraspanins and platelet markers that may reflect OA pathogenesis. The present study investigated the associations between these EV surface markers and articular cartilage degradation, subjectively and objectively assessed pain, and functional limitations in primary knee OA (KOA). Serum EVs were determined by high-sensitivity flow cytometry (large CD61+ EVs) and single particle interferometric reflectance imaging sensor (small CD41+, CD63+, CD81+, and CD9+ EVs) from end-stage KOA patients and controls (n = 8 per group). Knee pain and physical functions were assessed with several health- and pain-related questionnaires, established measurements of physical medicine, and neuromuscular examination. The obtained data were analyzed using supervised and unsupervised univariate and multivariate models. With the combined dataset of cartilage thickness, knee function, pain, sensation, and EV molecular signatures, we identified highly correlated groups of variables and found several EV markers that were statistically significant predictors of pain, physical limitations, and other aspects of well-being for KOA patients, for instance CD41+/CD63+/CD9+ small EVs associated with the range of motion of the knee, physical performance, and pain sensitivity. Particular serum EV subpopulations showed clear associations with KOA pain and functional limitations, suggesting that their implications in OA pathophysiology warrant further study.

Predictive Role of Platelet and Inflammation Markers for Severe COVID-19 by Propensity Score Matching.

This study aims to ascertain the predictive value of platelet and inflammation markers in severe cases of COVID-19. A retrospective real-world cohort study was conducted using propensity score matching (PSM). Patients were classified into severe and non-severe COVID-19 groups based on the severity of the disease, and the correlation between severe COVID-19 and laboratory parameters at admission was analyzed. The study included 397 adult patients, comprising 212 (53%) males and 185 (47%) females. Among these, 309 were non-severe and 88 were severe cases. The severe group had a higher median age than the non-severe group (60 vs. 42 years, p < 0.001). Independent risk factors for severe COVID-19 included age, diabetes comorbidity, fever, respiratory symptoms, platelet count, high-sensitivity C-reactive protein (hsCRP), interleukin-6 (IL-6), and the ratio of arterial oxygen partial pressure (PaO2) to the fraction of inspired oxygen (FiO2) (P/F ratio). After one-to-one PSM, adjusted for age, diabetes comorbidities, fever, and respiratory symptoms, significant differences in laboratory parameters at admission were observed. Compared to the non-severe group (n = 71), in the severe group (n = 71), elevated levels of hsCRP (median: 27.1 mg/L vs. 14.6 mg/L, p = 0.005) and IL-6 (median: 16.2 pg/mL vs. 15.3 pg/mL, p = 0.005) were observed, while platelet count (164 ± 36 × 109 vs. 180 ± 50 × 109, p = 0.02) and P/F ratio (median: 351 vs. 397, p = 0.001) were reduced. Elevated levels of hsCRP and IL-6, along with reduced platelet count and P/F ratio at admission, were significantly associated with severe COVID-19 and may serve as predictive indicators.

Effectiveness of D-dimer in predicting distant metastasis in colorectal cancer.

Patients with cancer often present with a hypercoagulable state, which is closely associated with tumor progression. The purpose of this study was to assess the diagnostic efficacy of D-dimer in predicting distant metastasis in colorectal cancer (CRC). This study included 529 patients diagnosed with CRC at our hospital between January 2020 and December 2022. Plasma coagulation indicators and tumor markers were collected prior to treatment and their diagnostic efficacy for predicting CRC metastasis was assessed by receiver operating characteristic (ROC) curves. Independent risk factors for evaluating tumor metastasis were obtained by multivariate logistic regression analysis. The level of D-dimer in the metastatic group was significantly higher than that in the non-metastatic group (P<0.001). The results of the multiple logistic regression analysis indicated that lower level of prealbumin and platelet, and higher level of glucose, CEA and D-dimer were independent risk factors for distant metastasis in patients with CRC (P<0.05, respectively). The combination of prealbumin, glucose, D-dimer, platelet and tumor markers (PRE2) was found to be significantly more effective in predicting metastasis of CRC when compared to the combination of tumor marker alone (PRE1, P<0.001). Plasma D-dimer may be a novel tumor marker for screening metastases of CRC.