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Sickle Cell Disease (SCD) is a severe genetic disorder causing vascular occlusion and pain by upregulating the adhesion molecule P-selectin on endothelial cells and platelets. It primarily affects infants and children, causing chronic pain, circulatory problems, organ damage, and complications. Thus, effective treatment and management are crucial to reduce SCD-related risks. Anti-P-selectin antibody Crizanlizumab (Crimab) has been used to treat SCD. In this study, the heavy and light chain (HC and LC) genes of anti-P-Selectin antibody Crimab were cloned into a plant expression binary vector. The HC gene was under control of the duplicated 35S promoter and nopaline synthase (NOS) terminator, whereas the LC gene was under control of the potato proteinase inhibitor II (PIN2) promoter and PIN2 terminator. Agrobacterium tumefaciens LBA4404 was used to transfer the genes into the tobacco (Nicotiana tabacum cv. Xanthi) plant. In plants the genomic PCR and western blot confirmed gene presence and expression of HC and LC Crimab proteins in the plant, respectively. Crimab was successfully purified from transgenic plant leaf using protein A affinity chromatography. In ELISA, plant-derived Crimab (CrimabP) had similar binding activity to P-selectin compared to mammalian-derived Crimab (CrimabM). In surface plasmon resonance, the KD (dissociation binding constant) and response unit values were lower and higher than CrimabP, respectively. Taken together, these results demonstrate that the transgenic plant can be applied to produce biofunctional therapeutic monoclonal antibody. [ABSTRACT FROM AUTHOR]

Background: Virus co-infection or reactivation may modify the host response during cerebral malaria. Cytomegalovirus (CMV) DNAemia has been associated with increased morbidity and mortality in adults with sepsis; however, the impact of CMV DNAemia on adverse outcomes in children with cerebral malaria is unknown. Methods: Clinical, physiological, and neurocognitive outcomes were compared in children aged 18 months to 12 years with cerebral malaria (N = 242) based on the presence or absence of CMV DNAemia 24 h after admission. The primary study outcome was subsequent in-hospital mortality. Secondary outcomes included the presence of acute kidney injury, neurocognitive impairment over a 2-year follow-up, and chronic kidney disease at the 1-year follow-up. Markers of platelet and endothelial cell activation and oxidative and nitrosative stress were measured to characterize the mechanisms by which CMV DNAemia might contribute to pathogenesis. Results: CMV DNAemia was present in 33 children with cerebral malaria (13.6%) 24 h after admission. CMV DNAemia was not significantly associated with mortality in this study. Children with CMV-DNAemia had a higher prevalence of acute kidney injury than those without CMV-DNAemia (59.4% vs. 38.6%, p = 0.03). There was no difference in the prevalence of chronic kidney disease or long-term neurocognitive impairment based on the presence of DNAemia. CMV DNAemia was associated with elevated plasma levels of P-selectin, angiopoietin-1, asymmetric dimethylarginine, and platelet counts. Conclusions: In children with cerebral malaria, CMV DNAemia is associated with acute kidney injury but not in-hospital mortality, chronic kidney disease, or long-term neurocognitive impairment. [ABSTRACT FROM AUTHOR]

Background/Objectives: The recommendations for antithrombotic therapy after transcatheter edge-to-edge mitral valve repair (TEER) are empirical, and the benefit of antiplatelet (APT) or anticoagulation therapy (ACT) remains undetermined. The study sought to investigate the degree and the timing of coagulation and platelet marker activation after TEER. Methods: This was a prospective study including 46 patients undergoing TEER. The markers of coagulation activation, namely prothrombin fragment 1 + 2 (F1 + 2) and thrombin-antithrombin III (TAT), and the markers of platelet activation, namely soluble P-Selectin and soluble CD-40 ligand (sCD40L), were measured at baseline, 24 h, 1 month, and 1 year after TEER. Results: At discharge, 20 (43%) patients received APT (single: 16, dual: 4), 24 (52%) received ACT, and 2 (4%) had both single APT and ACT. Levels of F1 + 2 and TAT significantly increased at 24 h post TEER (both p < 0.001), rapidly returning to baseline levels at 1 month. However, levels of F1 + 2 and TAT remained higher at 1 month in patients without ACT compared to patients with ACT (respectively, 303.1 vs. 148.1 pmol/L; p < 0.001 and 4.6 vs. 3.0 µg/L; p = 0.020), with a similar trend at 1 year. Levels of soluble P-selectin and sCD40L remained stable at all times after TEER (respectively, p = 0.071 and p = 0.056), regardless of the APT. Conclusions: TEER is associated with an acute activation of the coagulation system, with no increase in platelet activation markers. Hence, the use of dual APT is questionable in this population. Our results raise the hypothesis that the optimal antithrombotic therapy after TEER could be short-term ACT over APT. Further larger studies are warranted. [ABSTRACT FROM AUTHOR]

Background/Objectives: Leukocytes play a significant role in both acute kidney injury (AKI) and chronic kidney disease (CKD), contributing to pathogenesis and tissue damage. The process of leukocyte infiltration into the inflamed tissues is mediated by the interactions between the leukocytes and cell adhesion molecules (CAMs, i.e., E-selectin, P-selectin, and VCAM-1) present on the inner surface of the inflamed vasculature. Directly interfering with these interactions is a viable strategy to limit the extent of excessive inflammation; however, several small-molecule drug candidates failed during clinical translation. We hypothesized that a synthetic polymer presenting multiple copies of the high-affinity E-selecting binding peptide (P-Esbp) could block E-selectin-mediated functions and decrease leukocytes infiltration, thus reducing the extent of inflammatory kidney injury. Methods: P-Esbp was synthesized by conjugating E-selecting binding peptide (Esbp) to N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer with reactive ester groups via aminolysis. The effects of P-Esbp treatment on kidney injury were investigated in two different models: AKI model (renal ischemia—reperfusion injury—RIRI) and CKD model (adenine-induced kidney injury). Results: We found that the mRNA levels of E-selectin were up-regulated in the kidney following acute and chronic tissue injury. P-Esbp demonstrated an extended half-life time in the bloodstream, and the polymer accumulated significantly in the liver, lungs, and kidneys within 4 h post injection. Treatment with P-Esbp suppressed the up-regulation of E-selectin in mice with RIRI and attenuated the inflammatory process. In the adenine-induced CKD model, the use of the E-selectin blocking copolymer had little impact on the progression of kidney injury, owing to the compensating function of P-selectin and VCAM-1. Conclusion: Our findings provide valuable insights into the interconnection between CAMs and compensatory mechanisms in controlling leukocyte migration in AKI and CKD. The combination of multiple CAM blockers, given simultaneously, may provide protective effects for preventing excessive leukocyte infiltration and control renal injury. [ABSTRACT FROM AUTHOR]

Vascular endothelial cells are equipped with numerous specialized granules called Weibel-Palade bodies (WPBs). They contain a cocktail of proteins that can be rapidly secreted (3–5 min) into the vascular lumen after an appropriate stimulus such as thrombin. These proteins are ready without synthesis. Von Willebrand factor (VWF) and P-selectin are the main constituents of WPBs. Upon stimulation, release of ultralarge VWF multimers occurs and assembles into VWF strings on the apical side of endothelium. The VWF A1 domain becomes exposed in a shear-dependent manner recruiting and activating platelets. VWF is able to recruit leukocytes via direct leukocyte binding or via the activated platelets promoting NETosis. Ultralarge VWF strings are ultimately cleaved into smaller pieces by the protease ADAMTS-13 preventing excessive platelet adhesion. Under carefully performed flowing conditions and adequate dose of Shiga toxins, the toxin induces the release of ultralarge VWF multimers from cultured endothelial cells. This basic information allows insight into the pathogenesis of thrombotic thrombocytopenic purpura (TTP) and of STEC-HUS in the diarrhea phase. In TTP, ADAMTS-13 activity is deficient and systemic aggregation of platelets will occur after a second trigger. In STEC-HUS, stimulated release of WPB components in the diarrhea phase of the disease can be presumed to be the first hit in the damage of Gb3 positive endothelial cells. [ABSTRACT FROM AUTHOR]

Background and aims: Coronary obstruction following plaque rupture is a critical pathophysiological change in the progression of stable angina (SAP) to acute coronary syndrome (ACS). The accumulation of platelets and various inflammatory cells on apoptotic endothelial cells is a key factor in arterial obstruction after plaque rupture. Through single-cell sequencing analysis (scRNA-seq) of plaques from SAP and ACS patients, we identified significant changes in the annexin V and P-selectin glycoprotein ligand 1 pathways. Staphylococcal superantigen-like 5 (SSL5) is an optimal antagonist P-selectin glycoprotein ligand 1 (PSGL1), while annexin V (AnxA5) can precisely detect dead cells in vivo. We constructed the SSL5-AnxA5 fusion protein and observed its role in preventing the interaction between apoptotic endothelial cells, platelets, and inflammatory cells. Methods: The scRNA-seq data were extracted from the Gene Expression Omnibus (GEO) database. Single-cell transcriptome analysis results and cell–cell communication were analyzed to identify the ACS and SAP cell clusters and elucidate the intercellular communication differences. Then, we constructed and verified a fusion protein comprising SSL5 and AnxA5 domains via polymerase chain reaction (PCR) and Western blot. The binding capacity of the fusion protein to P-selectin and apoptotic cells was evaluated by flow cytometry and AnxA5-FITC apoptosis detection kit, respectively. Furthermore, co-incubation and immunofluorescence allowed us to describe the mediation effect of it between inflammatory cells and endothelial cells or activated platelets. Results: Our analysis of the scRNA-seq data showed that SELPLG (PSGL1 gene) and ANNEXIN had higher information flowing in ACS compared to SAP. The SELPLG signaling pathway network demonstrated a higher number of interactions in ACS, while the ANNEXIN signaling pathway network revealed stronger signaling from macrophages toward monocytes in ACS compared to SAP. Competition binding experiments with P-selectin showed that SSL5-AnxA5 induced a decrease in the affinity of PSGL1. SSL5-AnxA5 effectively inhibited the combination of endothelial cells with inflammatory cells and the interaction of activated platelets with inflammatory cells. Additionally, this fusion protein exhibited remarkable capability in binding to apoptotic cells. Conclusions: The bifunctional protein SSL5-AnxA5 exhibits promising potential as a protective agent against local inflammation in arterial tissues, making it an excellent candidate for PSGL1-related therapeutic interventions. [ABSTRACT FROM AUTHOR]

Objective： To investigate the value of color Doppler ultrasound combined with serum chemokine C-C motif ligand 3(CCL3) and P-selectin levels for the diagnosis of postoperative lower extremity deep vein thrombosis(LEDVT) in patients with hip fractures. Methods： Eighty patients with hip fractures complicated with LEDVT admitted to Xi'an Gaoxin Hospital from September 2022 to September 2024 were regarded as the LEDVT group, 80 patients with hip fractures without LEDVT were regarded as the non-LEDVT group, and 80 healthy individuals who experienced physical examination were regarded as the control group. The levels of serum CCL3 and P-selectin in three groups were compared. Color Doppler flow imaging (CDFI) was applied to measure the inner diameters of the common femoral vein (CFV) and deep femoral vein (DFV), and the subcutaneous tissue thickening was calculated. Multivariate logistic regression was applied to analyze the influencing factors of LEDVT in patients with hip fractures. ROC curve was applied to analyze the diagnostic value of ultrasound combined with serum CCL3 and P-selectin levels for LEDVT in patients with hip fractures. Results： Compared to the control group, both serum CCL3 and P-selectin levels were significantly higher in the LEDVT group (P＜0.05). Compared to the non-LEDVT group, the levels of serum CCL3, P-selectin, CFV, DFV, and subcutaneous tissue thickening were prominently higher in the LEDVT group (P＜0.05). CFV, PFV, subcutaneous tissue thickening, CCL3, and P-selectin were influencing factors for LEDVT in patients with hip fractures (P＜0.05). The combined diagnostic AUC (0.956) was significantly higher than that of any single indicator, with a sensitivity of 81.25% and specificity of 96.25%. Conclusion：The levels of serum CCL3 and P-selectin are obviously upregulated in patients with hip fracture complicated with LEDVT, both of which are influencing factors for the occurrence of LEDVT in patients with hip fracture. Ultrasound combined with serum CCL3 and P-selectin levels has high diagnostic value for LEDVT in patients with hip fractures.

Yuhong Zhang, Jun Li Center for Obstetrics and Reproductive Medicine, The Affiliated Hospital of Yunnan University, Kunming City, Yunnan Province, 650032, People’s Republic of ChinaCorrespondence: Jun Li, Center for Obstetrics and Reproductive Medicine, The Affiliated Hospital of Yunnan University, No. 176, Qingnian Road, Kunming, Yunnan, 650032, People’s Republic of China, Email lijun1152061295@hotmail.comBackground: This study analyzed the expression and diagnostic value of hsa_circ_101209 in plasma of pregnant women with in deep vein thrombosis (DVT).Methods: By circRNA microarray detection and GO/KEGG analysis, hsa_circ_14797 targeting miRNA-mRNA network was predicted. Sixty women with DVT were selected as the DVT group, and 60 women without DVT as the non-DVT group. hsa_circ_14797 in plasma was detected, as well as D-dimer (D-D) concentration and P-selectin expression. Target genes that may be regulated by hsa_circ_14797 were predicted, and GO analysis and KEGG pathway enrichment analysis were performed.Results: hsa_circ_14797 was highly expressed in DVT. hsa_circ_14797 in plasma of DVT patients was positively correlated with D-D (r = 0.358, P = 0.001). The AUC of plasma hsa_circ_14797, D-D, and P-selectin for maternal DVT diagnosis were 0.787 (95% CI: 0.710– 0.864), 0.882 (95% CI: 0.821– 0.943), and 0.825 (95% CI: 0.754– 0.895), respectively. The AUC of hsa_circ_14797 combined with D-D was 0.886 (95% CI: 0.828– 0.944). The AUC of hsa_circ_14797 combined with P-selectin was 0.904 (95% CI: 0.853– 0.954). The AUC of P-selectin combined with D-D was 0.935 (95% CI: 0.893– 0.978). The AUC of hsa_circ_14797 combined with D-D and P-selectin was 0.953 (95% CI: 0.920– 0.986). The functions of hsa_circ_14797 included the biological processes of angiogenesis, vascular development, and vascular morphology. The enrichment pathways included PI3K-Akt pathway, TGF-β pathway, and cytokine-cytokine receptor interaction.Conclusion: hsa_circ_14797, D-D, and P-selectin in plasma of DVT pregnant patients are increased, and hsa_circ_14797 in plasma is positively correlated with D-D. hsa_circ_14797 combined with D-D and P-selectin can improve the accuracy of diagnosis of DVT and contribute to the early diagnosis of DVT.Keywords: Deep vein thrombosis, circRNA, D-dimer, P-selectin, Biomarkers

Our objective was to demonstrate the efficiency of therapy for antiphospholipid syndrome (APS) in pregnant women using plasmapheresis and intravenous immunoglobulins.92 women with diagnosed APS and 47 APS-free women with physiological pregnancy were under study. APS was diagnosed in accordance with Sydney Consensus Workshop (2006). Patients with APS were divided into groups depending on the method of treatment used: conventional therapy, plasmapheresis in addition to standard therapy, intravenous immunoglobulins (IVIG) in addition to standard therapy, and complex treatment, i.e., plasmapheresis and IVIG added to standard therapy. The levels of antiphospholipid antibodies, hemostasis parameters, P-selectin were measured in blood serum and plasma before and after treatment.Higher frequency of favorable pregnancy outcomes was shown in the group of patients with APS who were treated with combined therapy, i.e. in 96.3% of cases (term birth). Frequency of prematurity and fetal hypotrophy was significantly lower in the group of patients with APS treated with combined therapy. The most significant decrease in the level of antiphospholipid antibodies was observed in the group treated by plasmapheresis, IVIG and conventional therapy. Expression of P-selectin in the women with normal pregnancy without antiphospholipid antibodies, was significantly lower compared to pregnant women with APS.Usage of integrated approach using plasmapheresis, intravenous immunoglobulins and standard therapy is the most effective treatment for APS-related miscarriage. Implication of this strategy has reduced the incidence of obstetric complications and improved pregnancy outcomes due to increased frequency of term births, as well as lowest indices of pathology in the newborns. Analysis of P-selectin levels before and after treatment enables determination of the platelet activation levels, efficiency of therapeutic approaches, as well as medical drug correction of infavorable platelet hemostasis during therapy.

Kingsley Akaba,1 Edakabasi Akaba,2 Olukayode Oshatuyi,2 Brian Ssenkumba3 1Department of Haematology, University of Calabar, Calabar, Cross River State, Nigeria; 2Department of Pathology, University of Calabar Teaching Hospital, Calabar, Cross River State, Nigeria; 3Department of Pathology, Kampala International University Western Campus, Ishaka, UgandaCorrespondence: Kingsley Akaba, Department of Haematology, University of Calabar, Calabar, Cross-River State, Nigeria, Email akaba_kingsley@yahoo.comBackground: Venous thromboembolism is the second leading cause of mortality among cancer patients. The Khorana Risk Assessment Score (KRAS) is widely acknowledged as the most validated tool in this context.Aim: To assess the thrombotic risk in cancer patients using the modified Khorana Risk Assessment Score, examine the association between modified KRAS and soluble P-selectin levels, and document the utilization of thromboprophylaxis among cancer patients at the University of Calabar Teaching Hospital.Methods: This was a cross-sectional hospital-based recruiting 100 cancer patients. Seven millilitres of blood were collected for complete blood count and P-selectin assay. Continuous variables were expressed as mean and standard deviation, while categorical variables were summarized using frequencies. Chi-square was employed to compare VTE risk status across genders, different cancer types, and guideline compliance. The significance level was set at 0.05.Results: Participants age ranged from 19 to 87 years, with a male-to-female ratio of 1:1.6. The most common female cancer was Breast at 40.32% and prostate cancer at 65.79% was the most common in males. Seventy nine percent and 21% of participants had intermediate and high-risk modified KRAS scores respectively. The median level of soluble P-selectin among cancer patients was 23.00 within the interquartile range. Significant associations were observed between cancer types and sex, VTE risk assessment and cancer types, and cancer types and risk score.Conclusion: The risk of VTE among cancer patients ranges from intermediate to high, going by the modified Khorana risk score irrespective of the P selectin level, with underutilization of thromboprophylaxis. There is little adherence to the Khorana score in our setting, hence the need for greater application and knowledge of this predictive score in clinical practice to improve outcomes and quality of life.Keywords: thrombosis, cancer, P-selectin

Abstract Background Activation of endothelial cells and platelets in atherothrombosis is characterized by upregulation of P-selectin. As a consequence, P-selectin represents a potential target for molecular imaging to identify thrombosis at an early stage. Fucoidan is a polysaccharide ligand extracted from brown algae with nanomolar affinity for P-selectin. This first-in-human study evaluated in healthy volunteers the safety, whole-body biodistribution, and dosimetry of 99mTc-fucoidan (Good Manufacturing Practices grade). We also investigated whether we could observe binding of 99mTc-fucoidan to human thrombi ex vivo and in vivo. In ten healthy volunteers, conjugate whole-body scans were performed up to 24 h following intravenous injection of 99mTc-fucoidan (370 MBq). Moreover, 99mTc-fucoidan uptake in ex vivo human thrombi (n = 11) was measured by gamma counting. Additionally, three patients with a newly diagnosed deep vein thrombosis (DVT) were subjected to 99mTc-fucoidan SPECT/CT imaging. Results 99mTc-fucoidan was well tolerated in all participants without any drug-related adverse events. The total-body absorbed dose in males was comparable to females (0.012 ± 0.004 vs. 0.011 ± 0.005 mSv/MBq; p = 0.97). Gamma counting experiments demonstrated binding of tracer to ex vivo human thrombi that was 16% lower after blocking with a natural P-selectin ligand, Sialyl Lewis X. 99mTc-fucoidan demonstrated specific uptake at the thrombus site in one out of three scanned patients with DVT. Conclusions 99mTc-Fucoidan has a favorable biodistribution and safety profile. 99mTc-fucoidan exhibited specific binding to human thrombi in both in vivo and ex vivo settings. Nonetheless, the in vivo results do not support further clinical investigation of 99mTc-fucoidan as an imaging modality for DVT. Other clinical implementations of a technetium− 99m-labeled P-selectin tracer should be considered. Trial registration: Clinicaltrials,NCT03422055.Registered 01/15/2018. URL: https://clinicaltrials.gov/study/NCT03422055 Landelijk Trial Register, NL7739. Registered 4/2/2019 . https://onderzoekmetmensen.nl/en/trial/26785

Background: Antithrombotic medications, including antiplatelet agents, are standard treatments for patients with hyperlipidemia who have a high risk of developing cardiovascular disease (CVD). The ongoing exploration of new antiplatelet agents with minimal bleeding effects is crucial, including the investigation of potential compounds derived from natural products. This study intended to evaluate the antiplatelet effects of a combined extract of sappan wood (Caesalpinia sappan L.) and red ginger (Zingiber officinale var. Rubrum) in high‐fat diet‐(HFD)‐induced rats. Methods: Eighteen male Wistar rats were grouped into six groups (n = 3): control, negative, positive, and three groups of various combinations of extracts. All groups, excluding the control group, were fed an HFD for 8 weeks. In the eighth week, the control and negative groups were given carboxyl methyl cellulose (CMC) 0.5%, the positive control group was administered aspirin, and the other three groups were administered the combination extract of sappan wood and red ginger at various doses for 2 weeks. Blood samples were collected to assess the levels of hyperlipidemia and platelet hyperactivity markers by enzyme‐linked immunosorbent assay (ELISA). The physiological effects of platelet hyperactivity were evaluated using the tail bleeding assay. Results: HFD‐induced hypertriglyceridemia and hypercholesterolemia synergistically enhanced platelet hyperactivity after 8 weeks of induction. Interestingly, administration of all doses of the combined extract for 2 weeks significantly decreased the platelet activation markers P‐selectin, RANTES, and PCSK9 in a dose‐dependent manner compared with the negative control. In addition, the combination of sappan wood and red ginger extract at dose 3 (sappan wood:red ginger: 200:800 mg/200 bw/day) significantly extended the bleeding time of rats (p < 0.05) compared to the negative control. Conclusion: Collectively, our results highlight the antiplatelet effect of a combination of sappan wood and red ginger extract in HFD‐fed rats. [ABSTRACT FROM AUTHOR]

Ovarian cancer (OC) presents daunting lethality rates worldwide, with frequent late-stage diagnosis and chemoresistance, highlighting the need for improved prognostic approaches. Venous thromboembolism (VTE), a major cancer mortality factor, is partially driven by endothelial dysfunction (ED). ED's pro-inflammatory state fosters tumour progression, suggesting a VTE-independent link between ED and cancer. Given this triad's interplay, ED markers may influence OC behaviour and patients' prognosis. Thus, the impact of ED-related genes and single-nucleotide polymorphisms (SNPs) on OC-related VTE and patient thrombogenesis-independent prognosis was investigated. NOS3 upregulation was linked to lower VTE incidence (χ2, p = 0.013), while SELP upregulation was associated with shorter overall survival (log-rank test, p = 0.048). Dismissing patients with VTE before OC diagnosis, SELP rs6136 T allele carriers presented lower progression-free survival (log-rank test, p = 0.038). Nevertheless, due to the SNP minor allele underrepresentation, further investigation is required. Taken together, ED markers seem to exhibit roles that depend on the clinical context, such as tumour-related thrombogenesis or cancer prognosis. Validation with larger cohorts and more in-depth functional studies are needed for data clarification and potential therapeutic strategies exploitation to tackle cancer progression and thrombosis in OC patients. [ABSTRACT FROM AUTHOR]

Systemic lupus erythematosus (SLE) is a heterogeneous autoimmune disease characterized by severe organ damage and lacking curative treatment. While various immune cell types, especially dysfunctional B and T cells and neutrophils, have been related with disease pathogenesis, limited research has focused on the role of monocytes in SLE. Increased DNA extracellular traps, apoptosis and necrosis have been related to lupus pathogenesis. Our goal is to analyze the contribution of P-selectin glycoprotein ligand 1 (PSGL-1) in SLE monocytes to disease pathogenesis by investigating the control exerted by PSGL-1 on monocyte apoptosis and DNA extrusion in extracellular traps (METs). Monocytes from active disease patients (aSLE) exhibited reduced levels of PSGL-1. Importantly, lower PSGL-1 levels in SLE monocytes associated with several clinical characteristics, including anti-dsDNA autoantibodies, lupus anticoagulant, clinical lung involvement, and anemia. Monocytes from SLE patients showed higher susceptibility to apoptosis than healthy donors (HD) monocytes and PSGL-1/P-selectin interaction decreased secondary necrosis in HD but not in aSLE monocytes. Regarding METs, aSLE monocytes exhibited higher susceptibility to generate METs than HD monocytes. The interaction of HD monocytes with P-selectin induced Syk activation and reduced the levels of DNA extruded in METs. However, in aSLE monocytes, PSGL-1/P-selectin interaction did not activate Syk or reduce the amount of extruded DNA. Our data suggest a dysfunctional PSGL-1/P-selectin axis in aSLE monocytes, unable to reduce secondary necrosis or the amount of DNA released into the extracellular medium in METs, potentially contributing to lupus pathogenesis. [ABSTRACT FROM AUTHOR]

Background: Prostaglandins are naturally occurring local mediators that can participate in the modulation of the cardiovascular system through their interaction with Gs/Gi-coupled receptors in different tissues and cells, including platelets. Thrombin is one of the most important factors that regulates platelet reactivity and coagulation. Clinical trials have consistently shown that omega-3 fatty acid supplementation lowers the risk for cardiovascular mortality and morbidity. Since omega-3 fatty acids are the main precursors of PGE3 in vivo, it would be relevant to investigate the effects of PGE3 on Thrombin Receptor Activating Peptide (TRAP-6)-induced platelet reactivity to determine the receptors and possible mechanisms of action of these compounds. Methods: We have measured platelet aggregation, P-selectin expression, and vasodilator-stimulated phosphoprotein (VASP) phosphorylation to evaluate platelet reactivity induced by TRAP-6 to determine the effects of PGE3 on platelet function. Results: We assessed the ability of DG-041, a selective prostanoid EP3 receptor antagonist, and of ONO-AE3-208, a selective prostanoid EP4 receptor antagonist, to modify the effects of PGE3. PGE3 inhibited TRAP-6-induced platelet aggregation and activation. This inhibition was enhanced in the presence of a Gi-coupled EP3 receptor antagonist and abolished in the presence of a Gs-coupled EP4 receptor antagonist. The effects of PGE3 were directly related to changes in cAMP, assessed by VASP phosphorylation. Conclusions: The general effects of PGE3 on human platelet reactivity are the consequence of a balance between activatory and inhibitory effects at receptors that have contrary effects on adenylate cyclase. These results indicate a potential mechanism by which omega-3 fatty acids underlie cardioprotective effects. [ABSTRACT FROM AUTHOR]

Background: Activation of endothelial cells and platelets in atherothrombosis is characterized by upregulation of P-selectin. As a consequence, P-selectin represents a potential target for molecular imaging to identify thrombosis at an early stage. Fucoidan is a polysaccharide ligand extracted from brown algae with nanomolar affinity for P-selectin. This first-in-human study evaluated in healthy volunteers the safety, whole-body biodistribution, and dosimetry of 99mTc-fucoidan (Good Manufacturing Practices grade). We also investigated whether we could observe binding of 99mTc-fucoidan to human thrombi ex vivo and in vivo. In ten healthy volunteers, conjugate whole-body scans were performed up to 24 h following intravenous injection of 99mTc-fucoidan (370 MBq). Moreover, 99mTc-fucoidan uptake in ex vivo human thrombi (n = 11) was measured by gamma counting. Additionally, three patients with a newly diagnosed deep vein thrombosis (DVT) were subjected to 99mTc-fucoidan SPECT/CT imaging. Results: 99mTc-fucoidan was well tolerated in all participants without any drug-related adverse events. The total-body absorbed dose in males was comparable to females (0.012 ± 0.004 vs. 0.011 ± 0.005 mSv/MBq; p = 0.97). Gamma counting experiments demonstrated binding of tracer to ex vivo human thrombi that was 16% lower after blocking with a natural P-selectin ligand, Sialyl Lewis X. 99mTc-fucoidan demonstrated specific uptake at the thrombus site in one out of three scanned patients with DVT. Conclusions: 99mTc-Fucoidan has a favorable biodistribution and safety profile. 99mTc-fucoidan exhibited specific binding to human thrombi in both in vivo and ex vivo settings. Nonetheless, the in vivo results do not support further clinical investigation of 99mTc-fucoidan as an imaging modality for DVT. Other clinical implementations of a technetium− 99m-labeled P-selectin tracer should be considered. Trial registration: Clinicaltrials,NCT03422055.Registered 01/15/2018. URL: https://clinicaltrials.gov/study/NCT03422055Landelijk Trial Register, NL7739. Registered 4/2/2019. https://onderzoekmetmensen.nl/en/trial/26785 [ABSTRACT FROM AUTHOR]

Objectives: A number of reports on the role of selectin in the process of carcinogenesis, at the stage of proliferation and metastasis, have been available. The aim of the study was to analyze (s)P- and (s)L-selectin serum concentrations in women with EC and to compare these concentrations to clinical/pathological parameters and disease progression using surgical-pathological staging data. Material and methods: A total of 46 patients with EC and 50 healthy controls were included in the study. Serum concentrations of sL- and sP-selectins were measured in all participants. The oncologic protocol was implemented in all women from the study group. Results: Significantly higher serum concentrations were found in EC women as compared to controls. No statistically significant differences were found between the concentrations of the soluble forms of selectins and the following parameters: histologic type of EC, histologic tumor differentiation, depth of myometrial infiltration, cervical involvement, distant metastases, vascular space invasion, and disease advancement. Slightly higher (s)P-selectin concentrations were observed in serous carcinoma, in women with cervical involvement, in the sera of women with vascular space invasion and with advanced stages of the disease. Slightly higher mean (s)P-selectin concentrations correlated with lower differentiation of the tumor. Slightly higher mean (s)P-selectin concentration was detected in the sera of women with lymph node metastases and with the serosal and/or adnexal involvement. The results were statistically insignificant, but they almost reached statistical significance. Conclusions: L- and P-selectins play a role in the biology of EC. The absence of an unambiguous relationship between differences in (s)L- and (s)P-selectin levels and disease advancement suggests that they do not play a vital role in tumor progression in endometrial cancer. [ABSTRACT FROM AUTHOR]

Increased platelet activity is a risk factor of thrombotic events in cardiovascular patients. We studied the relationship between platelet function, platelet size, and the content of reticulated platelets (RP) in patients with coronary heart disease (CHD, n = 55) and acute coronary syndrome (ACS, n = 95) receiving acetylsalicylic acid + clopidogrel or ticagrelor, respectively. The control group consisted of patients with risk factors for CHD, but with no CHD/ACS and free of antiplatelet drugs (n = 66). Platelet function was evaluated by the exposure of activated glycoprotein (GP) IIb-IIIa and P-selectin. In the control group, platelets were activated by TRAP (Thrombin Receptor Activating Peptide) 10 µM, and ADP 20, 5, 2.5 µM, and in the CHD/ACS groups, by TRAP 10 µM, and ADP 20 5 µM (±epinephrine 20 µM). Platelet size was assessed by the mean volume, % large forms, and forward scattering. RP were stained by thiazole orange. In the control group, activated GP IIb-IIIa and P-selectin correlated with platelet size and RP content after platelet activation by all agonists. Despite the decrease in platelet activity by antiplatelet drugs, most correlations (primarily for activated GP IIb-IIIa) were preserved in the CHD/ACS patients. In conclusion, increased platelet size and RP content are associated with increased platelet activity and the reduced efficacy of antiplatelet therapy. [ABSTRACT FROM AUTHOR]

Integrins were originally identified as receptors for extracellular matrix (ECM) and cell-surface molecules (e.g., VCAM-1 and ICAM-1). Later, we discovered that many soluble growth factors/cytokines bind to integrins and play a critical role in growth factor/cytokine signaling (growth factor-integrin crosstalk). We performed a virtual screening of protein data bank (PDB) using docking simulations with the integrin headpiece as a target. We showed that several growth factors (e.g., FGF1 and IGF1) induce a integrin-growth factor-cognate receptor ternary complex on the surface. Growth factor/cytokine mutants defective in integrin binding were defective in signaling functions and act as antagonists of growth factor signaling. Unexpectedly, several growth factor/cytokines activated integrins by binding to the allosteric site (site 2) in the integrin headpiece, which is distinct from the classical ligand (RGD)-binding site (site 1). Since 25-hydroxycholesterol, a major inflammatory mediator, binds to site 2, activates integrins, and induces inflammatory signaling (e.g., IL-6 and TNFα secretion), it has been proposed that site 2 is involved in inflammatory signaling. We showed that several inflammatory factors (CX3CL1, CXCL12, CCL5, sPLA2-IIA, and P-selectin) bind to site 2 and activate integrins. We propose that site 2 is involved in the pro-inflammatory action of these proteins and a potential therapeutic target. It has been well-established that platelet integrin αIIbβ3 is activated by signals from the inside of platelets induced by platelet agonists (inside-out signaling). In addition to the canonical inside-out signaling, we showed that αIIbβ3 can be allosterically activated by inflammatory cytokines/chemokines that are stored in platelet granules (e.g., CCL5, CXCL12) in the absence of inside-out signaling (e.g., soluble integrins in cell-free conditions). Thus, the allosteric activation may be involved in αIIbβ3 activation, platelet aggregation, and thrombosis. Inhibitory chemokine PF4 (CXCL4) binds to site 2 but did not activate integrins, Unexpectedly, we found that PF4/anti-PF4 complex was able to activate integrins, indicating that the anti-PF4 antibody changed the phenotype of PF4 from inhibitory to inflammatory. Since autoantibodies to PF4 are detected in vaccine-induced thrombocytopenic thrombosis (VIPP) and autoimmune diseases (e.g., SLE, and rheumatoid arthritis), we propose that this phenomenon is related to the pathogenesis of these diseases. P-selectin is known to bind exclusively to glycans (e.g., sLex) and involved in cell-cell interaction by binding to PSGL-1 (CD62P glycoprotein ligand-1). Unexpectedly, through docking simulation, we discovered that the P-selectin C-type lectin domain functions as an integrin ligand. It is interesting that no one has studied whether P-selectin binds to integrins in the last few decades. The integrin-binding site and glycan-binding site were close but distinct. Also, P-selectin lectin domain bound to site 2 and allosterically activated integrins.

Traumatic brain injury (TBI) is characterized by heterogeneity in terms of injury severity, mechanism, outcome, and pathophysiology. A single biomarker alone is unlikely to capture the heterogeneity of even one injury subtype, necessitating the use of panels of biomarkers. Herein, we focus on traumatic cerebrovascular injury and investigate associations of a panel of 16 vascular injury-related biomarkers with indices of TBI severity and outcomes using data from 159 participants in the Transforming Research and Clinical Knowledge in TBI (TRACK-TBI) Pilot Study. Associations of individual biomarkers and clusters of biomarkers identified using non-linear principal components analysis with TBI severity and outcomes were assessed using logistic regression models and Spearmans correlations. As individual biomarkers, higher levels of thrombomodulin, angiopoietin (Ang)-2, von Willebrand factor, and P-selectin were associated with more severe injury; higher levels of Ang-1, Tie2, vascular endothelial growth factor (VEGF)-C, and basic fibroblast growth factor (bFGF) were associated with less severe injury (all p  0.05). In conclusion, in this trauma-center based population of acute TBI patients, biomarkers of microvascular injury were associated with TBI severity.

BACKGROUND: COVID-19 has been associated with endothelial injury, resultant microvascular inflammation and thrombosis. Activated endothelial cells release and express P-selectin and von Willebrand factor, both of which are elevated in severe COVID-19 and may be implicated in the disease pathophysiology. We hypothesized that crizanlizumab, a humanized monoclonal antibody to P-selectin, would reduce morbidity and death in patients hospitalized for COVID-19. METHODS: An international, adaptive, randomized controlled platform trial, funded by the National Heart, Lung, and Blood Institute, randomly assigned 422 patients hospitalized with COVID-19 with moderate or severe illness to receive either a single infusion of the P-selectin inhibitor crizanlizumab (at a dose of 5 mg/kg) plus standard of care or standard of care alone in an open-label 1:1 ratio. The primary outcome was organ support-free days, evaluated on an ordinal scale consisting of the number of days alive free of organ support through the first 21 days after trial entry. RESULTS: The study was stopped for futility by the data safety monitoring committee. Among 421 randomized patients with known 21-day outcomes, 163 patients (77%) randomized to the crizanlizumab plus standard-of-care arm did not require any respiratory or cardiovascular organ support compared with 169 (80%) in the standard-of-care-alone arm. The adjusted odds ratio for the effect of crizanlizumab on organ support-free days was 0.70 (95% CI, 0.43-1.16), where an odds ratio >1 indicates treatment benefit, yielding a posterior probability of futility (odds ratio 1] = 0.879). CONCLUSIONS: Crizanlizumab, a P-selectin inhibitor, did not result in improvement in organ support-free days in patients hospitalized with COVID-19. REGISTRATION: URL: https://www. CLINICALTRIALS: gov; Unique identifier: NCT04505774.

Recognition of integrins by CD62P has not been reported and this motivated a docking simulation using integrin αvβ3 as a target. We predicted that the C-type lectin domain of CD62P functions as a potential integrin ligand and observed that it specifically bound to soluble β3 and β1 integrins. Known inhibitors of the interaction between CD62P-PSGL-1 did not suppress the binding, whereas the disintegrin domain of ADAM-15, a known integrin ligand, suppressed recognition by the lectin domain. Furthermore, an R16E/K17E mutation in the predicted integrin-binding interface located outside of the glycan-binding site within the lectin domain, strongly inhibited CD62P binding to integrins. In contrast, the E88D mutation that strongly disrupts glycan binding only slightly affected CD62P-integrin recognition, indicating that the glycan and integrin-binding sites are distinct. Notably, the lectin domain allosterically activated integrins by binding to the allosteric site 2. We conclude that CD62P-integrin binding may function to promote a diverse set of cell-cell adhesive interactions given that β3 and β1 integrins are more widely expressed than PSGL-1 that is limited to leukocytes.

Abstract Objectives The purpose of this study was to demonstrate the discriminating predictive indicators in peripheral blood and left atrium blood for predicting the risk of left atrial spontaneous echo contrast (LASEC) in atrial fibrillation patients underwent catheter ablation. Methods A total of 108 consecutive AF patients treated with radiofrequency ablation between July 2022 and July 2023 were enrolled and divided into two groups based on preprocedural transesophageal echocardiography: the non LASEC group (n = 71) and the LASEC group (n = 37). Circulating platelet and endothelial- derived MPs (PMPs and EMPs) in peripheral blood and left atrial blood were detected. Plasma soluble P-selectin (sP-selectin) and von Willebrand factor (vWF) were observed. Diagnostic efficiency was measured using receiver operating characteristic (ROC) curve. Results Peripheral sP-selectin, vWF and EMPs expressions elevated in all subjects when compared to those in left atrium blood. Levels of sP-selectin and vWF were significantly higher in peripheral blood of LASEC group than those of non LASEC group (p = 0.0018,p = 0.0271). Significant accumulations of peripheral PMPs and EMPs were documented in LASEC group by comparison with non LASEC group (p = 0.0395,p = 0.018). The area under curve(AUC) of combined PMPs and sP-selectin in predicting LASEC was 0.769 (95%CI: 0.678–0.845, sensitivity: 86.49%, specificity: 59.15%), significantly larger than PMPs or sP-selectin alone. Conclusions Expressions of PMPs, sP-selectin, EMPs and vWF Increased in NVAF patients with LASEC and that might be potential biomarkers for LASEC prediction.

Objectives: The purpose of this study was to demonstrate the discriminating predictive indicators in peripheral blood and left atrium blood for predicting the risk of left atrial spontaneous echo contrast (LASEC) in atrial fibrillation patients underwent catheter ablation. Methods: A total of 108 consecutive AF patients treated with radiofrequency ablation between July 2022 and July 2023 were enrolled and divided into two groups based on preprocedural transesophageal echocardiography: the non LASEC group (n = 71) and the LASEC group (n = 37). Circulating platelet and endothelial- derived MPs (PMPs and EMPs) in peripheral blood and left atrial blood were detected. Plasma soluble P-selectin (sP-selectin) and von Willebrand factor (vWF) were observed. Diagnostic efficiency was measured using receiver operating characteristic (ROC) curve. Results: Peripheral sP-selectin, vWF and EMPs expressions elevated in all subjects when compared to those in left atrium blood. Levels of sP-selectin and vWF were significantly higher in peripheral blood of LASEC group than those of non LASEC group (p = 0.0018,p = 0.0271). Significant accumulations of peripheral PMPs and EMPs were documented in LASEC group by comparison with non LASEC group (p = 0.0395,p = 0.018). The area under curve(AUC) of combined PMPs and sP-selectin in predicting LASEC was 0.769 (95%CI: 0.678–0.845, sensitivity: 86.49%, specificity: 59.15%), significantly larger than PMPs or sP-selectin alone. Conclusions: Expressions of PMPs, sP-selectin, EMPs and vWF Increased in NVAF patients with LASEC and that might be potential biomarkers for LASEC prediction. [ABSTRACT FROM AUTHOR]

Introduction Thirdhand smoke (THS) is associated with many public health and disease concerns, such as respiratory illness, cancer, lipidemia, and cardiovascular disease (CVD). We have previously shown that a moderate to long-term exposure to THS increases the risk of thrombosis. However, whether short-term exposure to THS would produce any effects remains to be discovered. Therefore, this study investigated the impact of 1-month THS exposure on platelet function, in vivo and in vitro, and on cytokine response, in a sex-dependent manner. Aims and Methods Secondhand smoke or clean air (CA) exposed upholstery materials for 1 week were kept in cages housed with 5–6 mice, and the procedure was repeated for 4 weeks. These THS-exposed mice were evaluated for thrombogenesis and platelet function assays. In addition, cytokines expression was evaluated from pooled serum. Results Compared to the CA group, THS exposure significantly shortened the tail bleeding time and carotid artery thrombus formation. Moreover, the female mice appeared more sensitive to THS exposure than males. Furthermore, platelet aggregation, dense granule secretion, and P-selectin activation markers were significantly elevated due to THS exposure. In addition, high-throughput screening showed at least 30 cytokines differentially modulated by THS in females relative to 26 in male mice. Conclusions Collectively, these results demonstrate that 1 month of THS exposure represents a high health risk, in part, by triggering a prothrombotic phenotype that appears to be more significant in females, who are at a much higher risk for occlusive CVD. Additionally, changes in cytokine levels mediate some of the THS-induced occlusive effects. Implications This study revealed that THS exposure for 1 month is detrimental to the cardiovascular health of both sexes; however, females could be more aggressively affected than males. In addition, interleukins and chemokines could be critical factors for initiating prothrombotic activity due to THS exposure. [ABSTRACT FROM AUTHOR]

BACKGROUND: Although plateletpheresis donation is commonly accepted as a safe procedure, its influence on platelet function, coagulation system and fibrinolysis is not completely elucidated. OBJECTIVES: In this study, we tried to assess the effects of plateletpheresis on donor's hemostasis system by measuring platelet activation, development of platelet-leukocyte aggregates, and coagulation activation. STUDY DESIGN: Prospective observational study. METHODS: We used flow cytometry to determine the levels of platelet-monocyte complexes (PMC) and platelet-neutrophil complexes (PNC). sP-selectin and prothrombin fragment (PF) 1 + 2 values were determined by ELISA. RESULTS: The PMC levels increased significantly seven days after apheresis in comparison with just after apheresis and 24 h after apheresis (p < 0.05). The PNC levels increased significantly seven days after apheresis compared to immediately after apheresis (p < 0.05). sP-selectin values decreased significantly immediately after apheresis (p < 0.05). While sP-selectin values increased seven days after apheresis in comparison with immediately after apheresis and 24 h after apheresis, but there were not statistically significant differences for sP-selectin levels (p > 0.05). PF1 + 2 levels decreased significantly immediately after apheresis compared to pre-apheresis (p < 0.05) and increased 24 h after apheresis and seven days after apheresis, but these differences were not statistically significant. CONCLUSION: We concluded that plateletpheresis affects platelet activation but does not cause any change in coagulation activation. [ABSTRACT FROM AUTHOR]

This study investigated the roles of P‐selectin and Clara cell secretory protein 16 (CC16) levels in the pathogenesis of severe adenovirus (ADV) pneumonia in children and evaluated their ability to predict disease. Fifty‐one children (age, 1–5 years) with ADV pneumonia who were admitted to Xiamen Children's Hospital were included in this study and divided into the mild group (24 patients) and severe group (27 patients). A control group comprising healthy children of the same age who underwent routine physical examinations during the same period (30 patients) was also included. The univariate analysis demonstrated that the levels of the white blood cell count and C‐reactive protein, procalcitonin, d‐dimer, and P‐selectin were increased in a severe group compared with a mild group, while CC16 levels were significantly decreased (p < 0.05). The logistic regression analysis revealed that P‐selectin and CC16 levels were independent risk factors for severe ADV pneumonia in children. The areas under the ROC curves suggested that P‐selectin and CC16 exhibited high predictive value for severe ADV pneumonia. P‐selectin values more than 898.58 pg/mL and CC16 values less than 11.355 ng/mL predicted severe ADV pneumonia. P‐selectin and CC16 levels are correlated with the severity of ADV pneumonia in children. [ABSTRACT FROM AUTHOR]

Cardiovascular diseases generate high costs for public health systems, making it necessary to develop/discover anticoagulant or antiplatelet agents. In this context, the present study investigated the antithrombotic action of the essential oil of Levisticum officinale W.D.J. Koch in an in vitro experimental model. The oil chemical composition was characterized using gas chromatography coupled to mass spectrometry (GC–MS). The trypan blue exclusion assay in platelets (in vitro) was carried out. Systemic toxicity was studied by an in vivo model of Galleria mellonella larvae. Platelet aggregation assay was performed by turbidimetry in an aggregometer device, using the adenosine receptor agonist, ADP, and epinephrine (EPI). Coagulation was determined by prothrombin activity time (PT) and partially activated thromboplastin time (aPTT) in the coagulation system. Platelet activation was assessed by P-selectin expression and reactive oxygen species (ROS) content by flow cytometry and fluorescence, respectively. The oil is composed of monoterpenes, including α-terpinyl acetate, β-phellandrene, β-myrcene, α-terpineol, and (Z)-β-ocimene. The oil did not show toxicity in vitro and in vivo. The oil significantly reduced platelet aggregation against ADP and EPI. EPI-induced aggregation was most potently inhibited. Treatment of platelets with oil showed a decrease in P-selectin expression and ROS production when compared to vehicle. On the other hand, the oil did not interfere in PT and aPTT. The results demonstrated that the essential oil of Levisticum officinale has antiplatelet action but without anticoagulant effects. [ABSTRACT FROM AUTHOR]

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 24 h 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. [ABSTRACT FROM AUTHOR]

Studies conducted on animal models have identified several therapeutic targets for myelofibrosis, the most severe of the myeloproliferative neoplasms. Unfortunately, many of the drugs which were effective in pre-clinical settings had modest efficacy when tested in the clinic. This discrepancy suggests that treatment for this disease requires combination therapies. To rationalize possible combinations, the efficacy in the Gata1low model of drugs currently used for these patients (the JAK1/2 inhibitor Ruxolitinib) was compared with that of drugs targeting other abnormalities, such as p27kip1 (Aplidin), TGF-β (SB431542, inhibiting ALK5 downstream to transforming growth factor beta (TGF-β) signaling and TGF-β trap AVID200), P-selectin (RB40.34), and CXCL1 (Reparixin, inhibiting the CXCL1 receptors CXCR1/2). The comparison was carried out by expressing the endpoints, which had either already been published or had been retrospectively obtained for this study, as the fold change of the values in the corresponding vehicles. In this model, only Ruxolitinib was found to decrease spleen size, only Aplidin and SB431542/AVID200 increased platelet counts, and with the exception of AVID200, all the inhibitors reduced fibrosis and microvessel density. The greatest effects were exerted by Reparixin, which also reduced TGF-β content. None of the drugs reduced osteopetrosis. These results suggest that future therapies for myelofibrosis should consider combining JAK1/2 inhibitors with drugs targeting hematopoietic stem cells (p27Kip1) or the pro-inflammatory milieu (TGF-β or CXCL1). [ABSTRACT FROM AUTHOR]

In recent years, substantial advancements have been made in understanding the pathophysiology of ischemic stroke. Despite these developments, therapeutic options for cerebral ischemia remain limited due to stringent time windows and various contraindications. Consequently, there has been a concentrated effort to elucidate the underlying mechanisms of cerebral ischemic injury. Emerging research indicates that neutrophil extracellular traps (NETs) exacerbate inflammation and damage in ischemic brain tissue, contributing to neuronal cell death. The inhibition of NETs has shown potential in preventing thrombosis and the infiltration of immune cells. Central to the formation of NETs are P-selectin and its ligand, P-selectin glycoprotein ligand-1 (PSGL-1), which represent promising therapeutic targets. This review explores the detrimental impact of P-selectin, PSGL-1, and NETs on cerebral ischemia. Additionally, it delineates the processes by which P-selectin and PSGL-1 stimulate NETs production and provides evidence that blocking these molecules reduces NETs formation. This novel insight highlights a potential therapeutic avenue that warrants further investigation by researchers in the field. [ABSTRACT FROM AUTHOR]

Introduction: The role and function of P-selectin levels in various inflammatory and immune-mediated diseases have been established. Whether they have an association with inflammatory skin diseases such as vitiligo and psoriasis needs to be established. Objective: The objective of this study was to assess P-selectin levels in psoriasis and vitiligo and to compare them with matched controls without skin disease. Materials and Methods: The study included a total of 90 subjects with age- and sex-matched – 30 each in psoriasis, vitiligo and 30 controls without skin disease. Psoriasis and vitiligo severity was assessed using the Psoriasis Area and Severity Index and the Vitiligo Area Scoring Index scores. P-selectin levels were assessed and compared among the groups. P-selectin levels were also compared with the severity of psoriasis and vitiligo. Chi-square and analysis of variance tests were used to compare the data. Results: The mean age of subjects was 36.28 ± 11.80 years. Majority of the subjects were males (65.6%). The three groups were matched for age, sex, and other demographics. The mean P-selectin levels were 610.43 ± 134.19, 292.52 ± 60.99, and 158.97 ± 34.76 ng/ml, respectively, in the psoriasis, vitiligo, and control groups, respectively (P < 0.001). No significant association of P-selectin levels was observed with psoriasis severity; however, with increasing vitiligo severity, there was a significant increase in P-selectin levels (P < 0.001). Conclusion: Patients with skin diseases have raised P-selectin levels. Within skin diseases, inflammatory diseases such as psoriasis have higher P-selectin levels as compared to autoimmune diseases such as vitiligo. A significant association of P-selectin levels was observed with vitiligo severity but not with psoriasis severity. [ABSTRACT FROM AUTHOR]

Introduction: Buerger's Disease (BD), or thromboangiitis obliterans (TAO), is a vasculoocclusive disorder of unknown etiology. Cigarette smoking is considered to be the main risk factor for the development of TAO. This study aimed to assess the activation of platelets in patients with BD compared to a control group. Materials and methods: Thirty patients (24 men and six women, median age 51.4 years) with TAO were included in the study. In the control group there were 20 healthy adults (16 men and four women, median age 44.0 years). Platelet activation was measured by light aggregometry and flow cytometry methods with and without platelet activators, such as thrombin receptor agonist peptide (TRAP-6) and adenosine diphosphate (ADP). Results: The velocity and intensity of aggregation increased in patients with TAO (p <0.05). MFI (mean fluorescence intensity) of P-selectin, CD63 (GP53-component of lysosomal membrane), and PAC-1 (platelet activation complex-1) on platelets increased in patients (p <0.05). Significantly higher aggregation velocities were noted in smoking patients in response to collagen (Coll) and to arachidonic acid (AA) compared to non-smoking ones. Aggregation velocity in response to AA was higher in heavy smokers compared to all smokers. A significantly higher intensity of aggregation was found in moderate smokers in response to epinephrine (EPI) and AA compared to non-smoking ones. A positive correlation between cholesterol concentrations and its fractions (LGL, TG), and platelet aggregation and the MFI of platelets, was found in patients with an increased cholesterol level. The aggregation velocity with EPI and AA was negatively correlated with HDL level. [ABSTRACT FROM AUTHOR]

Background: Studies regarding hypercoagulation in Non-Arteritic Anterior Ischemic Optic Neuropathy (NAION) patients have produced conflicting results. With a presumption that the early coagulation phase may affect the occurrence of NAION, this study aims to investigate the early coagulation markers, E-selectin and P-selectin, to determine whether these biomolecular changes play a significant role in NAION, thus potentially leading to a better clinical approach. Methods: A cross-sectional study involving two groups of NAION subjects, a hypercoagulation group and a non-hypercoagulation group, was conducted in the Neuro-Ophthalmology Division, Department of Ophthalmology, FKUI-RSCM Kirana from October 2020 to April 2022. All patients were evaluated for E-selectin and P-selectin levels measured using flow cytometry. Results: A total of 42 subjects comprising 14 hypercoagulation and 28 non-hypercoagulation subjects were included. In all subjects, E-selectin was strongly correlated with P-selectin (r = 0.862, p < 0.001). There was no significant difference in E-selectin and P-selectin values between the groups (p = 0.317 for E-selectin, and p = 0.575 for P-selectin). Prothrombin time and international normalized ratio (INR) were inversely correlated with both E-selectin and P-selectin in the hypercoagulation group (p = 0.032, p = 0.030 for E-selectin and p = 0.044, p = 0.036 for P-selectin). There was no significant correlation between E-selectin and P-selectin for NAION-associated metabolic risk factors. However, higher E-selectin and P-selectin values were found in the presence of risk factors except for P-selectin in the hypertension group. Conclusion: This interesting finding opens up the potential for considering the involvement of E-selectin and P-selectin in the diagnostic strategy for NAION. It prompts consideration of whether assessing E-selectin and P-selectin levels should be recommended for all NAION patients. Furthermore, considering the role of E-selectin and P-selectin in the early coagulation process, future studies are also needed to further evaluate whether anticoagulants could play a role in the choice of treatment for NAION despite a clinically hypercoagulable state.

The combination of inflammation and thrombosis is a hallmark of many cardiovascular diseases. Under such conditions, platelets are recruited to an area of inflammation by forming platelet-leukocyte aggregates via interaction of PSGL-1 on leukocytes and P-selectin on activated platelets, which can bind to the endothelium. While particulate drug carriers have been utilized to passively redirect leukocytes from areas of inflammation, the downstream impact of these carriers on platelet accumulation in thromboinflammatory conditions has yet to be studied. Here, we explore the ability of polymeric particles to divert platelets away from inflamed blood vessels both in vitro and in vivo. We find that untargeted and targeted micron-sized polymeric particles can successfully reduce platelet adhesion to an inflamed endothelial monolayer in vitro in blood flow systems and in vivo in a lipopolysaccharide-induced, systemic inflammation murine model. Our data represent initial work in developing cargo-free, anti-platelet therapeutics specifically for conditions of thromboinflammation.

Objective To analyze the changes in important indicators such as P-selectin, vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF), and pH value in platelet-rich plasma (PRP) after short-term cryopreservation at -80 ℃, and to provide a theoretical basis for the low-temperature storage of PRP. Methods Each of PRP samples prepared from the blood from 30 healthy donors was divided into 5 aliquots, with 0.5 mL retained per aliquot. These samples were de-signated as group A (fresh sample) and groups B to E (cryopreservation for 5, 10, 15, and 20 d, respectively). The pH values of the five groups were measured. The concentrations of P-selectin, PDGF, and VEGF in each group were determined by the ELISA method. The effect of groups A and E on the vascular formation capacity of human umbilical vein endothelial cells (HUVEC) was determined by the angiogenesis assay. Results No significant differences were observed in the concentrations of P-selectin, PDGF, and VEGF as well as the pH values between group A and groups B to E (P >0.05). Compared with the fresh sample (group A), the number of vessels formed from HUVECs in the sample frozen for 20 d (group E) was basically the same, with no significant differences (P>0.05). Conclusion The cryopreservation of PRP at -80 ℃ has no significant effects on its pH value and concentrations of P-selectin, VEGF, and PDGF, thus extending the clinical application time of PRP.

BackgroundFeminizing hormonal therapy (FHT) and HIV potentially alter cardiovascular disease (CVD) risk in transgender women (TW).MethodsTW were enrolled in Los Angeles, California and Houston, Texas and frequency-matched to Multicenter AIDS Cohort Study cisgender men (CM) on age, race, substance use, and abacavir use. Biomarkers of CVD risk and inflammation were assessed via ELISA. Wilcoxon rank sum and Fisher's exact tests compared TW and CM. Multivariable linear regression assessed factors associated with biomarker concentrations.ResultsTW (HIV+ n  = 75, HIV- n  = 47) and CM (HIV+ n  = 40, HIV- n  = 40) had mean age 43-45 years; TW/CM were 90%/91% non-Hispanic Black, Hispanic, or Multiracial, 26%/53% obese, and 34%/24% current smokers; 67% of TW were on FHT. Among people with HIV (PWH), TW had higher median extracellular newly-identified receptor for advanced glycation end-products (EN-RAGE), lipoprotein-associated phospholipase A2 (LpPLA2), oxidized low-density lipoprotein (oxLDL), soluble tumor necrosis factor receptor type (sTNFR) I/II, interleukin (IL)-8 and plasminogen activator inhibitor (PAI)-1, but lower soluble CD14, von Willebrand factor (vWF) and endothelin (ET)-1 levels than CM. Findings were similar for participants without HIV (all P

Donor-specific HLA Abs contribute to Ab-mediated rejection (AMR) by binding to HLA molecules on endothelial cells (ECs) and triggering intracellular signaling, leading to EC activation and leukocyte recruitment. The molecular mechanisms involving donor-specific HLA Ab-mediated EC activation and leukocyte recruitment remain incompletely understood. In this study, we determined whether TLRs act as coreceptors for HLA class I (HLA I) in ECs. We found that human aortic ECs express TLR3, TLR4, TLR6, and TLR10, but only TLR4 was detected on the EC surface. Consequently, we performed coimmunoprecipitation experiments to examine complex formation between HLA I and TLR4. Stimulation of human ECs with HLA Ab increased the amount of complex formation between HLA I and TLR4. Reciprocal coimmunoprecipitation with a TLR4 Ab confirmed that the crosslinking of HLA I increased complex formation between TLR4 and HLA I. Knockdown of TLR4 or MyD88 with small interfering RNAs inhibited HLA I Ab-stimulated P-selectin expression, von Willebrand factor release, and monocyte recruitment on ECs. Our results show that TLR4 is a novel coreceptor for HLA I to stimulate monocyte recruitment on activated ECs. Taken together with our previous published results, we propose that HLA I molecules form two separate signaling complexes at the EC surface, that is, with TLR4 to upregulate P-selectin surface expression and capture of monocytes to human ECs and integrin β4 to induce mTOR-dependent firm monocyte adhesion via ICAM-1 clustering on ECs, two processes implicated in Ab-mediated rejection.

Activated platelets (PLT) could adhere to tumor cells to promote tumor progression and metastasis. Therefore, inhibiting platelet activation and blocking the interaction between tumor cells and platelets are an effective approach to inhibit distal tumor metastasis. In this study, chondroitin sulfate (CS)–glycyrrhetinic acid (GA) dual ligand-modified liposomes (CS–GA–LIP) were prepared for codelivery of aspirin (ASP) and curcumin (CUR). In order to simulate the tumor microenvironment, we established the "HuH-7 + PLT" in vitro coculture cell model and the "H22 + NIH/3T3" coinjected mice model. Physicochemical analysis showed that the particle size of CUR&ASP/CS–GA–LIP was 133.27 nm with high stability. In vitro studies found that CUR&ASP/CS–GA–LIP could target both hepatocellular carcinoma cells and platelets, enhance drug retention in tumor cells, and effectively inhibit tumor proliferation and migration. In addition, in vivo antitumor results demonstrated that CUR&ASP/CS–GA–LIP could aggregate in tumor tissues, inhibit platelet activation and neoangiogenesis, and down-regulate the expression of matrix metalloproteinase 2 (MMP-2), exhibiting strong antiproliferative and antipulmonary metastatic abilities. Consequently, the combination therapy based on CS–GA–LIP might be a valuable antihepatocellular carcinoma strategy. [ABSTRACT FROM AUTHOR]

This review examines the endothelial glycocalyx's role in inflammation and explores its involvement in coagulation. The glycocalyx, composed of proteins and glycosaminoglycans, interacts with von Willebrand Factor and could play a crucial role in anchoring it to the endothelium. In inflammatory conditions, glycocalyx degradation may leave P-selectin as the only attachment point for von Willebrand Factor, potentially leading to uncontrolled release of ultralong von Willebrand Factor in the bulk flow in a shear stress-dependent manner. Identifying specific glycocalyx glycosaminoglycan interactions with von Willebrand Factor and P-selectin can offer insights into unexplored coagulation mechanisms. [ABSTRACT FROM AUTHOR]

Objectives Detecting occult cancer in patients with unprovoked venous thromboembolism (VTE) remains a significant challenge. Our objective was to investigate the potential predictive role of coagulation-related biomarkers in the diagnosis of occult malignancies. Methods We conducted a nested case-control study with a 1-year prospective cohort of 214 patients with unprovoked VTE, with a focus on identifying occult cancer. At the time of VTE diagnosis, we measured various biomarkers, including soluble P-selectin (sP-selectin), dimerized plasmin fragment D (D-dimer), platelets, leukocytes, hemoglobin, total extracellular vesicles (EVs), EVs expressing tissue factor on their surface (TF+EVs), and EVs expressing P-selectin on their surface (Psel+EVs) in all participants. Results We observed statistically significant increased levels of sP-selectin (P =.015) in patients with occult cancer. Despite an increase in Psel+EVs, TF+EVs, D-dimer, and platelets within this group, however, no significant differences were found. When sP-selectin exceeded 62 ng/mL and D-dimer surpassed 10,000 µg/L, the diagnosis of occult cancer demonstrated a specificity of up to 91% (95% CI, 79.9%-96.7%). Conclusions The combination of sP-selectin and D-dimer can be a valuable biomarker in detecting occult cancer in patients with unprovoked VTE. Further research is necessary to ascertain whether easily measurable biomarkers such as sP-selectin and D-dimer can effectively distinguish between patients who have VTE with and without hidden malignancies. [ABSTRACT FROM AUTHOR]

Background: Venous Thrombo Embolism (VTE) is the third most common cause of vascular mortality worldwide and comprises Deep-Vein Thrombosis (DVT) and Pulmonary Embolism (PE). It is also the second most common cause of death among cancer patients. VTE contributes immensely to the morbidity and mortality of cancer patients, with a life threaten PE being 3 times more common in cancer patients compared to non-cancer patients. Cancer patients are said to have a 5 to 7 fold increased risk of developing VTE. P-selectin can serve as a promising marker which could help to increase the precision of predicting the risk of VTE among cancer patients when combine with established validated tools. Aim: This study seeks to unravel the increasing trend of cancer associated VTE and the intricate and multifaceted role of P-selectin in cancer associated VTE. Method: A vigorous literature search was performed via the internet search engines linked to academic databases including PubMed, Google Scholar, Ebsco, Hinari, Scopus, etc. Conclusion: This review underscores the dynamic of cancer associated VTE and context-dependent role of P-selectin in cancer associated VTE respectively. As we poke into the relationship between cancer, VTE and P-selectin, as this will create an insight for therapeutic intervention. [ABSTRACT FROM AUTHOR]

Purpose: To investigate the effect of combining aspirin with clopidogrel on high-sensitivity C-reactive protein (hs-CRP) and P-selectin in patients with acute coronary syndrome (ACS). Methods: The medical records of 198 ACS patients treated at Zhaotong Traditional Chinese Medicine Hospital, Zhaotong, China between January 2022 and January 2023 were analyzed retrospectively. Control group (88 patients) received aspirin (300 mg once, and subsequently a maintenance dose of 100 mg daily for 6 months), while the study group (118 patients) received the same dose of aspirin and clopidogrel (150 mg once, and a maintenance dose of 75 mg daily for 6 months). Levels of P-selectin, interleukin-6 (IL-6), hs-CRP, IL-8, tumor necrosis factor-α (TNF-α), efficacy, and incidence of adverse reactions in both groups were determined. Results: There was no significant difference in P-selectin and hs-CRP levels between the two groups before treatment (p > 0.05). However, P-selectin and hs-CRP levels in the study group decreased significantly compared to control group (p < 0.0001). Similarly, there was no significant difference in IL-8, IL-6 and TNF-α levels between the two groups before treatment (p > 0.05). However, IL-8, IL-6, and TNF-α reduced significantly after treatment (p < 0.0001), with more significantly lower levels observed in the study group (p < 0.001). There was no significant difference in the incidence of adverse reactions between the two groups. Conclusion: Aspirin combined with clopidogrel significantly lowers P- selectin, and hs-CRP, IL-8, IL-6, and TNF-α levels without worsening adverse reactions. A more comprehensive analysis of the combined use of aspirin with clopidogrel including all the associated mechanisms in the treatment of ACS is necessary. [ABSTRACT FROM AUTHOR]