Endothelial Vascular Adhesion Molecule-1 Research Articles

Angiopoietin-2 is associated with sickle cell complications, including stroke risk, and decreases with hydroxyurea therapy

AbstractHydroxyurea reduces morbidity and mortality in children with sickle cell anemia (SCA). The endothelium is central to SCA-related complications including stroke. However, hydroxyurea’s impact on the endothelium is not well described. To address this gap, we measured plasma levels of endothelial activation markers (angiopoietin-2, P-selectin, soluble endothelial selectin [sE-selectin], soluble intercellular cellular adhesion molecule 1, and soluble vascular endothelial cellular adhesion molecule) by enzyme-linked immunosorbent assay after initiation of hydroxyurea therapy. Samples were collected from Ugandan children with SCA enrolled in a clinical trial evaluating hydroxyurea vs placebo (NOHARM trial). Samples were collected at enrollment; and then after 2, 4, and 12 months of follow-up. Longitudinal changes in biomarker levels were evaluated using linear mixed effects models. Transcranial Doppler (TCD) velocities were measured at 10 to 12 months follow-up to assess cerebral blood flow and primary stroke risk. Mediation analysis was used to explore causal pathways of hydroxyurea-mediated effects on TCD velocities. In total, 798 plasma samples were tested from 205 children (mean enrollment age, 2.2 years). At enrollment, higher levels of angiopoietin-2 were associated with a previous medical history of dactylitis, vaso-occlusive crises, acute chest syndrome, and transfusion (P < .05 for all). Hydroxyurea therapy at a fixed dose of 20 mg/kg per day decreased plasma angiopoietin-2, P-selectin, and sE-selectin levels over the study period (P < .05 for all). Angiopoietin-2 and sE-selectin were associated with higher TCD velocities. Mediation analysis suggests that hydroxyurea decreases TCD velocities through an increase in fetal and total hemoglobin. Increased fetal and total hemoglobin, and decreased white blood cell count may decrease TCD velocity, in part, through an angiopoiten-2–mediated pathway. This trial was registered at www.ClinicalTrials.gov as #NCT01976416.

Reduction of the risk of failure to achieve blood pressure control in patients with arterial hypertension and coronary heart disease: the role of quercetin

Aim. Determine the effect of quercetin on predictors of not reaching target blood pressure (BP) levels in patients with arterial hypertension (AH) and coronary heart disease (CHD). Materials and methods. 120 patients with stable coronary heart disease and hypertension of the II stage 2–3 degrees (66 women and 54 men) were examined. The patients were divided into 2 groups: Group I – 58 patients who, in addition to basic therapy (ramipril/amlodipine), received quercetin (Corvitin®), Group II – 62 patients who received only basic antihypertensive therapy. The parameters of daily blood pressure monitoring, blood lipid spectrum, C-reactive protein, the concentration of adhesion molecules (s-VCAM, s-ICAM-1), levels of endothelin-1 (ET-1), IL-1, IL-6, TNF-α were determined. Statistical processing of materials was carried out using the following programs: Minitab 21.0, StatPlus Pro 7.6.5, Wizard Pro, Jamovi, MedCalc. Results. We have established that the risk of not reaching the target level of daily SBP in the group of basic treatment of hypertension (group II) significantly increases with exceeding the level of LDL &gt;4.0 mmol/l (by 3.6 times), an increase in the level of IL-6 &gt;7 ng/ml (by 10 times), an increase in the level of TNF-α &gt;9 pg/ml (by 7 times), an increase in the level of VCAM &gt;1100 ng/ml (by 2.4 times). In the group of people who additionally took quercetin (group I), with an increase in the level of ET-1 &gt;1.1 pg/ml, the risk of failure to reach the target SBP increases by 2.6 times, in the II group – by 3.5 times toward to daily SBP and 8.6 times – toward to daily DBP. We found out that an increase in the concentration of IL-6 &gt; 7 ng/mg significantly increases the risk of arrhythmias by 6 times in group II. Conclusions. When quercetin is used, it is possible to achieve the target values of SBP (daily, daytime, nighttime) and DBP (daily, daytime, nighttime) in a significantly higher proportion of patients compared with those who are prescribed only standard antihypertensive therapy. Quercetin has an endothelial protective effect, as evidenced by a significant decrease in the levels of vascular endothelial adhesion molecules (s-VCAM), type I intercellular adhesion molecules (s-ICAM-1), ET-1, IL-1, IL-6, TNF-α. The additional use of quercetin against the background of standard antihypertensive therapy significantly reduces the risk of failure to achieve the target levels of SBP, DBP and the risk of heart rhythm disturbances even against the background of increased levels of proinflammatory cytokines and markers of endothelial dysfunction.

Probiotic-Fermented Camel Milk Attenuates Neurodegenerative Symptoms via SOX5/miR-218 Axis Orchestration in Mouse Models.

Multiple sclerosis is an autoimmune-mediated myelin damage disorder in the central nervous system that is widespread among neurological patients. It has been demonstrated that several genetic and epigenetic factors control autoimmune encephalomyelitis (EAE), a murine model of MS, through CD4+ T-cell population quantity. Alterations in the gut microbiota influence neuroprotectiveness via unexplored mechanisms. In this study, the ameliorative effect of Bacillus amyloliquefaciens fermented in camel milk (BEY) on an autoimmune-mediated neurodegenerative model using myelin oligodendrocyte glycoprotein/complete fraud adjuvant/pertussis toxin (MCP)-immunized C57BL6j mice is investigated. Anti-inflammatory activity was confirmed in the in vitro cell model, and inflammatory cytokines interleukins IL17 (from EAE 311 to BEY 227 pg/mL), IL6 (from EAE 103 to BEY 65 pg/mL), IFNγ (from EAE 423 to BEY 243 pg/mL) and TGFβ (from EAE 74 to BEY 133 pg/mL) were significantly reduced in BEY-treated mice. The epigenetic factor miR-218-5P was identified and confirmed its mRNA target SOX-5 using in silico tools and expression techniques, suggesting SOX5/miR-218-5p could serve as an exclusive diagnostic marker for MS. Furthermore, BEY improved the short-chain fatty acids, in particular butyrate (from 0.57 to 0.85 µM) and caproic (from 0.64 to 1.33 µM) acids, in the MCP mouse group. BEY treatment significantly regulated the expression of inflammatory transcripts in EAE mice and upregulated neuroprotective markers such as neurexin (from 0.65- to 1.22-fold) (p < 0.05), vascular endothelial adhesion molecules (from 0.41- to 0.76-fold) and myelin-binding protein (from 0.46- to 0.89-fold) (p < 0.03). These findings suggest that BEY could be a promising clinical approach for the curative treatment of neurodegenerative diseases and could promote the use of probiotic food as medicine.

The Effects of Pregnancy on the Pulmonary Immune Response in a Mouse Model of LPS-Induced Acute Lung Injury.

This study evaluated the effect of pregnancy on the pulmonary innate immune response in a mouse model of acute lung injury (ALI) using nebulized lipopolysaccharide (LPS). Pregnant (day 14) C57BL/6NCRL mice and nonpregnant controls received nebulized LPS for 15 minutes. Twenty-four hours later, mice were euthanized for tissue harvest. Analysis included blood and bronchoalveolar lavage fluid (BALF) differential cell counts, whole-lung inflammatory cytokine transcription levels by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), and whole-lung vascular cell adhesion molecule 1 (VCAM-1), intercellular adhesion molecule 1 (ICAM-1), and BALF albumin by western blot. Mature bone marrow neutrophils from uninjured pregnant and nonpregnant mice were examined for chemotactic response using a Boyden chamber and for cytokine response to LPS by RT-qPCR. In LPS-induced ALI, pregnant mice had higher BALF total cell (p < 0.001) and neutrophil counts (p < 0.001) as well as higher peripheral blood neutrophils (p < 0.01) than nonpregnant mice, but a similar increase (as compared with unexposed mice) in airspace albumin levels. Whole-lung expression of interleukin 6, tumor necrosis factor-α (TNF-α), and keratinocyte chemoattractant (CXCL1) was also similar. In vitro, marrow-derived neutrophils from pregnant and nonpregnant mice had similar chemotaxis to CXCL1 and N-formylmethionine-leucyl-phenylalanine, but neutrophils from pregnant mice expressed lower levels of TNF (p < 0.001) and CXCL1 (p < 0.01) after LPS stimulation. In uninjured mice, VCAM-1 was higher in lungs from pregnant versus nonpregnant mice (p < 0.05). In this model, pregnancy is associated with an augmented lung neutrophil response to ALI without increased capillary leak or whole-lung cytokine levels relative to the nonpregnant state. This may stem from increased peripheral blood neutrophil response and intrinsically increased expression of pulmonary vascular endothelial adhesion molecules. Differences in lung innate cell homeostasis may affect the response to inflammatory stimuli and explain severe lung disease in respiratory infection during pregnancy. · Inhalation of LPS in midgestation versus virgin mice is associated with increased neutrophilia.. · This occurs without a comparative increase in cytokine expression.. · This may be explained by pregnancy-enhanced pre-exposure expression of VCAM-1 and ICAM-1..

Avenanthramide-C Activates Nrf2/ARE Pathway and Inhibiting Ferroptosis Pathway to Improve Cognitive Dysfunction in Aging Rats.

Postoperative neurocognitive impairment (POCD) is a common complication after surgery and anesthesia, especially in elderly patients. Avenanthramide-C (AVC) test is a vascular endothelial cell adhesion molecule inhibitor with strong anti-inflammatory and antioxidant effects. The aim of this study was to investigate the effect and mechanism of AVC on POCD in aged rats to clarify the effect of AVC on POCD in aged rats. The aging rat model was established by continuous 200mg/kg propofol anesthesia. Repeated propofol anesthesia could severely impair spatial learning ability, memory and cognitive function, and could promote hippocampal apoptosis, oxidative stress injury, neuroinflammation and ferroptosis in aging rats. In addition, AVC not only improved cognitive dysfunction, but also significantly inhibited apoptosis, neuroinflammatory response, ferroptosis and oxidative stress level in the hippocampus of aging rats induced by repeated anesthesia. Further mechanistic studies manifested that the above protective effects of AVC on aging rats induced by repeated propofol anesthesia may be achieved by activating Nrf2/ARE pathway activity. AVC pretreatment has a preventive effect on cognitive dysfunction induced by repeated propofol anesthesia in aging rats, and the preventive effect of AVC may be realized by activating the Nrf2/ARE signaling pathway activity. Our results demonstrate that AVC preconditioning reduces postoperative neuronal loss and neuroinflammation, activates the Nrf2/ARE pathway, reduces oxidative stress injury, and improves POCD in aged rats.

Silymarin reduces retinal microvascular damage in streptozotocin-induced diabetic rats

Diabetic retinopathy is a severe microvascular problem in diabetes mellitus. Silymarin is a flavonoid compound, and according to previous studies, it is a bioactive compound with potent antioxidant and anti-inflammatory properties. This investigation aims to peruse the impact of silymarin against diabetic retinopathy in streptozotocin (STZ)-provoked rats. Thirty-two adult male Wistar rats were randomly allocated into the control group, STZ group, STZ + silymarin (50 mg/kg), and STZ + silymarin (100 mg/kg). STZ rats received silymarin every day until 2 months after diabetes induction. The serum and retinal tissues were collected 2 months after silymarin treatment to determine biochemical and molecular analyses. Silymarin markedly lowered the serum glucose concentration in diabetic rats. Silymarin reduced the increased levels of advanced glycosylated end products (AGEs), the receptors for AGEs (RAGE), and reactive oxygen species (ROS) in diabetic rats. Silymarin also attenuated the phosphorylation of p38 MAP kinase and nuclear factor (NF)-κB p65 and diminished diabetes-induced overexpression of inflammatory cytokines, vascular endothelial growth factor (VEGF), adhesion molecules, and extracellular matrix proteins in STZ rats. Our data suggested that silymarin has protective effects against diabetic retinopathy, which might be related to the inhibition of the AGEs/RAGE axis and its antioxidant and anti-inflammatory activities.

Study of type 1 vascular adhesion molecules in patients with acute myocardial infarction with ST segment elevation at different body weights

Objective: to study the level of adhesion molecules in patients with STEMI with different BMI at the hospital stage and 12 months after the index event.Materials and methods: the study included 126 people with STEMI who had undergone PCI and 27 people in the control group. The analysis of the level of svcam‑1 in peripheral blood at the beginning of the disease and after 12 months was carried out, BMI and waist volume were measured. An assessment of the nature and frequency of complications after STEMI was performed.Results: the levels of biomarkers of the vascular endothelial adhesion molecule type 1 increase during the acute period of stemi, statistically signifi tly decrease, but remain increased by 3.5 times 12 months after the index event compared with the initial values. There was no association of VCAM‑1 with visceral obesity in the groups of our patients. Vascular endothelial adhesion molecules of type 1 increase in patients with a fatal outcome, as well as with an increase in the severity of OSN and CHF. Thus, VCAM 1 can be a predictor of an unfavorable outcome of AMI.Conclusions: the article presents the study of a marker of systemic inflammation VCAM‑11 in patients with STEMI with various types of obesity or BMI at the stage of hospitalization and outpatient follow‑up during the year. The determination of the VCAM‑1 level can be used to assess the intensity of the inflammatory process and the risk of adverse outcomes.

Abstract 13231: Semaphorin3F Reduces Atherosclerosis by Decreasing Endothelial Activation and Barrier Permeability and Smooth Muscle Cell Migration

Coronary artery disease (CAD) remains a leading cause of death worldwide. To identify genes causally linked to CAD, we previously conducted large-scale genetic analyses, and identified an association between genes in the semaphorin pathway and CAD. Expression profiling in human atherosclerosis-relevant cell types and atherosclerotic plaques identified semaphorin3F (SEMA3F) as a possible candidate causally linked to atherosclerosis. Administration of SEMA3F reduced aortic lesion area in western-type diet-fed Ldlr -/- mice (Control: 12.1±1.4; SEMA3F: 8.1±0.8, % lesion area/total aortic sinus area, p=0.02). In contrast, neointimal formation was increased in Sema3f -/- mice with partial carotid artery ligation ( WT : 22.8±2.8; Sema3f -/- : 39.2±4.0; % intimal area/total vessel area, p=0.004). Additionally, SEMA3F administration increased re-endothelialization and reduced restenosis in Ldlr -/- mice that underwent carotid endothelial denudation (Control: 53.3±3.5; SEMA3F: 21.4±3.1; % intimal plaque area/vessel area, p=0.0006). Monocyte adhesion to aortic vascular endothelial cells (VECs) from Sema3f -/- mice was elevated, driven by increased NF-κB activation and increased expression of its target adhesion molecules, VCAM1 and ICAM1, in VECs. Monocyte transmigration through VECs from Sema3f -/- mice was also increased, likely due to increased membrane permeability, assessed by in-vivo intravital two-photon microscopy. The increased membrane permeability was modulated in part by decreased mTOR phosphorylation that in turn suppressed vascular endothelial cadherin protein expression. SEMA3F increased thick actomyosin filaments and elicited larger focal adhesions in VECs, suggesting decreased migration. In line with this, SEMA3F inhibited vascular smooth muscle cell (VSMC) migration and maintained the contractile phenotype of VSMCs, which was mediated by the Rho pathway. SEMA3F is causally atheroprotective in models of atherosclerosis and restenosis, mediated, in part, through the modulation of vascular endothelial cell permeability and adhesion molecule expression, and vascular smooth muscle cell migration and phenotype switching.

Study of markers of systemic inflammation (matrix metalloproteinase-9, vascular endothelial adhesion molecules of type 1) in patients with ST -segment elevation myocardial infarction at the hospital and outpatient stages

Aim. To study the level of MMP-9 and VCAM-1 in patients with AMI with ST-segment elevation at the hospital stage and one year after the index event, depending on the development of complications and changes in BMI and waist size (WS).&#x0D; Material and methods. The study included 126 people with STEMI after PCI, as well as 27 people in the control group. The level of MMP-9 and VCAM-1 in peripheral blood was analyzed. In addition to the standard methods of examination and diagnosis, BMI and WS were measured to identify groups with visceral obesity. The assessment of the frequency and nature of complications after STEMI was performed.&#x0D; Results. The levels of biomarkers of vascular endothelial adhesion molecule type 1 and matrix metalloproteinase type 9 are increased in the acute period of STEMI, statistically significantly reduced, but remain elevated 12 months after the index event, with VCAM-1 by 3.5 times, and MMP by almost 2 times compared to the initial values. The level of MMP-9 is significantly higher in excess body mass index and waist size, and VCAM-1 has no association with visceral obesity. Vascular endothelial adhesion molecules of type 1 and matrix metalloproteinases of type 9 are increased in patients with a fatal outcome, as well as with an increase in the severity of OSN and CHF. The level of matrix metalloproteinase has a strong relationship with fatal outcome and repeated MI, and the vascular endothelial adhesion molecule type 1 has a stronger relationship with the severity of CHF.&#x0D; Conclusion. The article studied markers of systemic inflammation (matrix metalloproteinase type 9 and vascular endothelial adhesion molecules type 1) in patients with ST-segment elevation myocardial infarction, depending on the presence of obesity at the hospital and outpatient stages. Simultaneous determination of MMP-9 and VCAM-1 levels can be used to assess the intensity of the inflammatory process and the risk of adverse outcomes.

Clinical Study on the Efficacy of Silver Ion Dressing Combined with Prontosan Gel Dressing in the Treatment of Diabetic Foot Ulcers and the Effect on Serum Inflammatory Factors.

Diabetic foot ulcers (DFUs) have a high disability rate and have a great impact on patients and society, and the search for effective and economical treatment options is a major clinical concern. In this study, 112 patients with DFU admitted to two hospitals from October 2018 to November 2020 were randomly divided into 56 cases each in the single group treated with Prontosan gel dressing and the joint group treated on silver ion dressing combined with Prontosan gel dressing. Both groups of patients were evaluated for efficacy after 30 days of treatment. The number of days for debridement, granulation tissue growth time, epithelial tissue formation time, and wound healing time were observed and recorded in both groups. The trauma area, visual analogue score (VAS), and levels of inflammatory factors such as vascular endothelial adhesion molecule-1 (VCAM-1), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and C-reactive protein (CRP) were recorded before and after treatment in both groups. The occurrence of adverse reactions such as edema, fever, infection, and rash during treatment was recorded in both groups for safety assessment. Comparison of the abovementioned data showed that the clinical efficacy of the joint group was significantly higher than that of the single group. The number of days to clear wounds, granulation tissue growth time, epithelial tissue formation time, and wound healing time were significantly lower in the joint group than in the single group. The trauma area, VAS score, VCAM-1, IL-6, TNF-α, and CRP levels decreased in both groups after treatment compared with the pretreatment levels, with the joint group being lower than the single group. The results also showed that the difference in the overall incidence of adverse reactions between the two groups was not statistically significant, and the incidence was low and transient. In addition to the usual treatment regimen of blood glucose control and improvement of microcirculation for patients with DFU, combined treatment with silver ionomer dressings and Prontosan gel dressings can promote ulcer healing and improve foot wound regression. It has a stronger antibacterial effect and can more effectively reduce the inflammatory response of the ulcerated surface with fewer adverse effects, making it an effective and safe method for the treatment of DFU, and has implications for promotion.

Therapeutic effect of ursolic acid on fetal development in pregnant rats with gestational diabetes mellitus via AGEs-RAGE signaling pathway.

To investigate the effect of ursolic acid on the fetal development of gestational diabetes mellitus (GDM) caused by streptozotocin (STZ) and explore the potential mechanism for it. For the current experimental research, SD rats (pregnant animal) were used. STZ has been used to cause the diabetes mellitus in pregnant rats. Rats with evolved GDM were randomly divided and ursolic acid was given to pregnant rats in the experimental period up to 19days in a dose-dependent manner. Blood samples and fetal rats of all group rats were collected at 19days (pregnant rats), fetal rats and placental rats were weighted and the blood glucose, plasma insulin, C-peptide, and lipid parameters of pregnant women were estimated prior to delivery. Advanced serum glycation end-products (AGEs) were estimated at regular intervals in the heart and brain of pregnant rats. Monocyte Chemoattractant Protein-1 (MCP-1), NADPH oxidase 2 (Nox2), Role of advanced glycation end product (RAGE), Vascular endothelial growth factor (VEGF), p65, and vascular cell adhesion molecule 1 (VCAM-1) mRNA expression were estimated in the placenta. STZ-induced GDM pregnant rats showed significantly decreased placental weight and weight of fetal rats and dose-dependent ursolic acid treatment (p<.001) improved placental weight and weight of fetal rats at dose-dependent levels. After the ursolic acid treatment, serum blood glucose and lipid level were improved especially fasting blood glucose (FBG), high density lipoprotein (HDL), hepatic glycogen, fasting insulin (FINS), and serum insulin level and reached near to the normal control group rats. The antioxidant level of pancreas and liver were significantly (p<.001) reduced by the dose-dependent treatment of ursolic acid. Treatment with Ursolic acid moderately but not significantly decreases the risk of fetal development defects relative to the GDM group. The potential effect on fetal development in Pregnant Rats with Gestational Diabetes Mellitus via AGEs-RAGE signaling pathway was shown by Ursolic acid. PRACTICAL APPLICATIONS: As we know that the gestational diabetes mellitus increases worldwide day by day. In the current experimental study, we try to examine the gestational diabetic effect of ursolic acid. The finding of the current study showed the gestational diabetic protective effect in the female rats via AGEs-RAGE signaling pathway. The result showed the antioxidant, anti-inflammatory, and biochemical parameters. On the basis of the result, we can say that the ursolic acid can be the protective drug for treatment of gestational diabetes mellitus.

Genetic Association of Clinical Complications for Genes Differentially Expressed in Reticulocytes of Sickle Cell Anemia

Introduction Sickle cell anemia (SCA) is characterized by hemolysis, a tissue hypoxia-driven increase in erythropoietin production, and a marked reticulocytosis reflecting stress erythropoiesis. Reticulocytes in SCA constitute 4-15% or more of peripheral red blood cells. Sickle reticulocytes express adhesion molecules that are normally cleared by exosome shedding, including α4β1 integrin, which is thought to contribute to vaso-occlusive crises by binding to vascular endothelial cell vascular cell adhesion molecule 1. Our recent study of the reticulocyte transcriptome in SCA versus healthy African Americans, using RNA-Seq, revealed differential genes that were implicated in a broad range of biological processes including terminal erythroid differentiation, the unfolded protein response, and altered membrane systems. In this study, we assessed the genetic association of these differential genes with disease complications relevant to sickle reticulocytes. Methods We analyzed vaso-occlusive complications including episodic severe pain and chronic avascular necrosis. We also analyzed hemolysis and anemia measured by a hemolysis index and hemoglobin concentration, respectively. The patient cohort combined 525 adolescent and adult patients from Walk PHaSST and 377 pediatric patients from PUSH. Genomic DNA isolated from peripheral blood mononuclear cells of Walk-PHaSST and PUSH patients were genotyped using Illumina Human 610-Quad SNP array and the genotypes were phased and imputed to 1000 genomes phase 3 data. Single nucleotide polymorphisms (SNPs) with minor allele frequency (MAF) &amp;gt;0.01 and imputation r2&amp;gt;0.7 were tested for gene-wise genetic association using SKAT. For severe pain and avascular necrosis, the covariates included age, gender, β globin genotype severity (HHbSS and HbSβ0-thalassemia versus HbSC and HbSβ+-thalassemia), and population stratification. For hemolysis and hemoglobin concentration, the covariates included age, gender, β globin genotype severity, alpha thalassemia, hydroxyurea treatment, recent blood transfusion, and population stratification. Results A total of 111 genes were analyzed. These genes were classified into 13 functional categories based on published research (transcription, post-transcription, translation, histone modification, cell cycle arrest, mitophagy, metabolism, iron homeostasis, antioxidative system, unfolded protein response, membrane skeleton and structural proteins, membrane adhesion molecules and blood group antigens, and membrane transporters). For each functional class, the number of significant gene-wise associations the functional class contains was compared to the number of significant associations from the other classes combined. Defining significance of gene-wise association at nominal P=0.05, we found that the "Unfolded protein response" contained more significant gene-wise associations with severe pain than the other functional classes (one-sided Fisher's exact test P=0.046), driven by genes FUCA1 (alpha-L- fucosidase) and CLN8 (associated with endoplasmic reticulum function). "Membrane skeleton and structural proteins" showed a trend of enrichment for hemolysis (P=0.057), driven by genes MYO5B and CD2AP, and for avascular necrosis (P=0.075), driven by genes MYLK, CAVIN2 and PALLD. Defining the significance of gene-wise association at a less stringent nominal P=0.1, "membrane adhesion molecules and blood group antigens" showed a trend of enrichment in association with severe pain (P=0.097), driven by genes GYPA, ITGA4 and ITGB1, the latter two encoding α4β1 integrin subunits which are abnormally present on the reticulocyte membrane and have been implicated in sickle vaso-occlusion. Regional scans revealed the SNPs with the most significant associations within these genes. Discussion Our results reveal enrichment of genetic association with vaso-occlusive and hemolytic complications in gene functional categories that showed altered expression in sickle reticulocytes. Future study is needed to validate these associations. Disclosures Gordeuk: CSL Behring: Consultancy, Research Funding; Global Blood Therapeutics: Consultancy, Research Funding; Imara: Research Funding; Ironwood: Research Funding; Novartis: Consultancy.

Daunorubicin associated to lipid core nanoparticles reduces atherosclerotic lesions, inflammation and cardiotoxicity in atherosclerosis rabbit model

Abstract Introduction Antiproliferative agents used in cancer chemotherapy have low toxicity and increased pharmacological action when carried into lipid core nanoparticles (LDE). LDE has similar structure to LDL and binds to LDL receptors through apo E it acquires in contact with plasma. Previously, we showed that treatment of rabbits with atherosclerosis with drugs such as taxanes and etoposide associate to LDE had pronounced reduction of the atherosclerotic lesions. In this study, we hypothesized whether daunorubicin (DNR) that belongs to another class of anti-cancer drugs, with a distinct mechanism of action and that has strong cardiotoxicity could also have effects against atherosclerosis. Methods Sixteen New Zealand male rabbits were fed with a 1% cholesterol diet for 8 weeks to induce atherosclerosis. After 4 weeks, the animals were treated weekly with LDE-DNR (6mg/kg, iv, n=9) or with LDE only (n=7). In addition, 3 animals without any intervention were used as a control group. Chow consumption, lipid and hematological profiles, body weight, and echocardiography were evaluated at baseline, pre-treatment and post-treatment. Morphometry and protein expression were performed to analyze the aortas. Results There was no difference in food intake and body weight for control, LDE and LDE-DNR groups. Total cholesterol, HDL-C, non-HDL-C and triglycerides increased in LDE and LDE-DNR groups when comparing baseline and post-treatment. Red blood cells decreased in LDE group at post-treatment in comparison to baseline, while LDE-DNR presented no such difference. Regarding aortic lesions of LDE-DNR group was 50% lower than LDE group. The protein expression of the inflammatory markers CD68, TNF-α and IL-6 in LDE-DNR was lower than LDE. Pro-apoptotic factors caspase 3, caspase 9 and BAX were also lower in LDE-DNR compared to LDE. Protein expression of vascular endothelial growth factor (VEGF) and vascular cell adhesion molecule (VCAM) was lower in the LDE-DNR group when compared to the LDE group. To test the cardiotoxicity of LDE-DNR, echocardiography was performed and observed that under LDE-DNR both systolic and diastolic function were preserved with no difference in cardiac mass in LDE-DNR when compared to control and LDE groups. However, LDE showed a significant increase in relative heart weight compared to control while LDE-DNR had no such difference. Treatment with LDE-DNR did not induce observable hematological toxicity. Conclusion The treatment with LDE-DNR reduced aortic lesions, diminished inflammatory and pro-apoptotic factors, diminished VCAM expression in the aorta and promoted cardiac preservation in a rabbit atherosclerosis model. In conclusion, LDE-DNR can be eligible for future developments in the quest for novel effective and safe treatments for atherosclerosis. Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): FAPESP (Fundaçao de Amparo a Pesquisa do Estado de Sao Paulo)

Sox17-mediated expression of adherent molecules is required for the maintenance of undifferentiated hematopoietic cluster formation in midgestation mouse embryos

Hematopoietic stem cell-containing intra-aortic hematopoietic cell clusters (IAHCs) emerge in the dorsal aorta of the aorta-gonad-mesonephros (AGM) region during midgestation mouse embryos. We previously showed that transduction of Sox17 in CD45lowc-Kithigh cells, which are one component of IAHCs, maintained the cluster formation and the undifferentiated state, but the mechanism of the cluster formation by Sox17 has not been clarified. By microarray gene expression analysis, we found that genes for vascular endothelial-cadherin (VE-cad) and endothelial cell-selective adhesion molecule (ESAM) were expressed at high levels in Sox17-transduced c-Kit+ cells. Here we show the functional role of these adhesion molecules in the formation of IAHCs and the maintenance of the undifferentiated state by in vitro experiments. We detected VE-cad and ESAM expression in endothelial cells of dorsal aorta and IAHCs in E10.5 embryos by whole mount immunohistochemistry. Cells with the middle expression level of VE-cad and the low expression level of ESAM had the highest colony-forming ability. Tamoxifen-dependent nuclear translocation of Sox17-ERT fusion protein induced the formation of cell clusters and the expression of Cdh5 (VE-cad) and ESAM genes. We showed the induction of the Cdh5 (VE-cad) and ESAM expression and the direct interaction of Sox17 with their promoter by luciferase assay and chromatin immunoprecipitation assay, respectively. Moreover, shRNA-mediated knockdown of either Cdh5 (VE-cad) or ESAM gene in Sox17-transduced cells decreased the multilineage-colony forming potential. These findings suggest that VE-cad and ESAM play an important role in the high hematopoietic activity of IAHCs and cluster formation.

Involvement of Caveolin-1-mediated transcytosis in the intratumoral accumulation of liposomes

For achieving efficient cancer treatment, it is important to elucidate the mechanism responsible for the accumulation of nanoparticles in tumor tissue. Recent studies suggest that nanoparticles are not delivered merely through gaps between tumor endothelial cells. We previously reported that the maturation of the vascular structure by the vascular endothelial cell growth factor receptor 2 (VEGFR2) using a previously developed siRNA delivery technology (RGD-MEND) significantly enhanced the accumulation of nanoparticles in types of cancers that area vessel-rich (renal cell carcinoma). This result was completely inconsistent with the generally accepted theory of the enhanced permeability and retention (EPR) effect. We hypothesized that a caveolin-1 (Cav1)-mediated transcellular route would be involved with the penetration of nanoparticles into tumor vasculature. To reveal the exact mechanism responsible for this enhancement, we observed the delivery of long-circulating liposomes (LPs) after Cav1 was co-suppressed by RGD-MEND with VEGFR2. The enhanced delivery of LPs by siRNA against VEGFR2 (siVEGFR2) was accompanied by the elevated expression of the Cav1 protein. In addition, Cav1 knockdown by siRNA against Cav1 (siCav1) canceled the enhanced delivery of LPs by siVEGFR2. The injection of siCav1 had no effect on the formation of alpha smooth muscle actin or vascular endothelial cell adhesion molecules. These results suggest that a Cav1-induced transcellular route and not a paracellular route, at least partially, contributes to the accumulation of nanoparticles in tumors.

Endothelial damage and circadian blood pressure profile in rheumatoid arthritis

To study the influence of the state of endothelium on the daily profile of arterial pressure (AP) in patients with rheumatoid arthritis (RA). In 70 RA pts carried out C-reactive protein (CRP), vascular endothelial adhesion molecule type 1 (sVCAM-1), antigen von Willebrand Factor (AG WF), interleukin-8 (Il-8), rheumatoid factor (RF), IgG, endotheline-1 (ET-1), number of desquamated endotheliocytes cells (DE), VS, activity of renin by immunoenzyme analysis. The dysfunction of endothelium was evaluated by calculation of DE. The functional methods included the daily monitoring of arterial pressure (AP). Arterial hypertension (AH) occurred in 40 (57.1%) pts. RA pts are revealed the signs of endothelial dysfunction, about which significant differences among the indices of activation of endothelium in comparison with control group testify. ET-1, sVCAM-1, vWF AG, Il-8, CRP content was higher in RA pts. Reliably above there was a number of DE. Reliable differences according to these indices depending of RA activity were discovered. With conducting of correlation analysis it is revealed, that markers of the activation of endothelium: sVCAM-1, vWF AG positively correlated with increasing RF IgG and indices of the immune inflammation: CRP, and DE number. In patients suffering from RA, showed signs of endothelial dysfunction. The positive correlation between endothelial damage and daily profile of AP show the relationship of these processes. Positive correlations between the damage of endothelium and disturbance of AP daily profile testify about the interrelation of these processes.

Fibronectin Costimulation of CD40L Expression Integrates Innate and Adaptive Inflammatory Signals

Abstract CD40 ligand (CD40L) is a well-known regulator of the adaptive immune response that induces and modulates the activation and differentiation of monocytes, dendritic cells, and lymphocytes. Less appreciated is its critical role in promoting the inflammatory response in CD40-expressing cells such as vascular endothelium. Expression of CD40L on TCR activated human CD4+ T cells is biphasic and upregulated by monocyte cell contact early and IL-2R signaling late. Here we demonstrate the existence of additional, distinct pathways that differentially regulate the expression of CD40L. We describe a novel link between vascular endothelial cell adhesion molecule 1 (VCAM-1), extracellular matrix (ECM) proteins, and CD40L expression on human CD4+T cells that constitutes an innate form of T cell costimulation modulated by the type and quantity of ECM protein present, with thrombospondin-1 inhibiting fibronectin-mediated costimulation of CD40L expression. Identification of this previously unrecognized β1 integrin pathway, which promotes CD40L expression on CD4+ T cells independently of monocytes and IL-2, provides new insights into our understanding of the potential role of CD40L in the establishment and progression of inflammatory disease.

Markers of endothelial dysfunction: pathogenetic role and diagnostic significance.

Endothelial dysfunction (ED) is considered one of the pathogenetic mechanisms of a whole range of diseases. Detection of specific biochemical markers in the blood is an effective way to ED diagnostics that characterize the vascular endothelium state. This review highlights the pathogenetic role of the factors synthesized by endotheliocytes whose level changes in biological fluids reflect violations of the endothelium basic physiological properties: vasomotor function, thromboresistance, angiogenesis regulation, barrier and adhesion functions. In particular, the participation of nitric oxide metabolites, asymmetric dimethylarginine, endothelin-1, metabolic products of arachidonic acid, von Willebrand factor, thrombomodulin, vascular endothelial growth factor, vasohibine-1 and adhesion molecules in the onset and development of ED are reviewed. The diagnostic significances of factors damaging endothelium, such as C-reactive protein, homocysteine and 8-hydroxy-2'-deoxyguanosine, are discussed. In addition, the literature data of recent years about the prospects of clinical implication the detection of the above-mentioned factors which indicates structural and functional endothelial cells damage are given. Particular attention is paid to the ED markers detection prognostic significance and the possibility of their practical use for the ED diagnosis. The search of literature for the current review was conducted in RSIC, CyberLeninka, Scopus, Web of Science, MedLine and PubMed databases from 2012 to 2018 using the following keywords: endothelial dysfunction, nitric oxide, asymmetric dimethylarginine, endothelin-1, prostacyclin, thromboxane A2, epoxyeicosatrienoic acids, von Willebrand factor, thrombomodulin, vascular endothelial growth factor, vasohibin-1, adhesive molecules, C-reactive protein, homocysteine, and 8-hydroxy-2-deoxyguanosine.

MARKERS OF ENDOTHELIAL DYSFUNCTION: PATHOGENETIC ROLE AND DIAGNOSTIC SIGNIFICANCE

Endothelial dysfunction (ED) is considered one of the pathogenetic mechanisms of a whole range of diseases. Detection of specific biochemical markers in the blood is an effective way to ED diagnostics that characterize the vascular endothelium state. This review highlights the pathogenetic role of the factors synthesized by endotheliocytes whose level changes in biological fluids reflect violations of the endothelium basic physiological properties: vasomotor function, thromboresistance, angiogenesis regulation, barrier and adhesion functions. In particular, the participation of nitric oxide metabolites, asymmetric dimethylarginine, endothelin-1, metabolic products of arachidonic acid, von Willebrand factor, thrombomodulin, vascular endothelial growth factor, vasohibine-1 and adhesion molecules in the onset and development of ED are reviewed. The diagnostic significances of factors damaging endothelium, such as C-reactive protein, homocysteine and 8-hydroxy-2’-deoxyguanosine, are discussed. In addition, the literature data of recent years about the prospects of clinical implication the detection of the above-mentioned factors which indicates structural and functional endothelial cells damage are given. Particular attention is paid to the ED markers detection prognostic significance and the possibility of their practical use for the ED diagnosis. The search of literature for the current review was conducted in RSIC, CyberLeninka, Scopus, Web of Science, MedLine and PubMed databases from 2012 to 2018 using the following keywords: endothelial dysfunction, nitric oxide, asymmetric dimethylarginine, endothelin-1, prostacyclin, thromboxane A2, epoxyeicosatrienoic acids, von Willebrand factor, thrombomodulin, vascular endothelial growth factor, vasohibin-1, adhesive molecules, C-reactive protein, homocysteine, and 8-hydroxy-2-deoxyguanosine.

Cerebral perfusion, arterial hypertension and endothelial dysfunction in rheumatoid arthritis

Seventy-eight patients with RA were divided into two groups: with- and without AH.The functional methods included ultrasonic duplex angioscanning and dopplerografy of the extracranial and intracranial arteries of the head and neck and daily 24 hour monitoring of arterial blood pressure (BP). C-reactive protein (CRP), vascular endothelial adhesion molecule type 1 (sVCAM-1), von Willebrand Factor antigen (vWF AG), interleukin-8 (Il-8), rheumatoid factor (RF) IgG, endothelin-1 (ET-1) were determined by immunoenzyme analysis and velocity of sedimentation (VS). The dysfunction of endothelium was evaluated by calculation of the number of desquamated endotheliocytes cells (DE). AH occurred in 46 (59%) patients. Cerebral hypoperfusion was observed in patients with RA in whom the reduction in the high-speed indices of blood flow were correlated with BP increase. There were negative correlations between the linear speed of blood flow on the common carotid arteries and average day and night systolic BP, average day and night diastolic BP, indices of time systolic BP and diastolic BP and avariability of BP. The same results were established for the intracranial arteries: inverse correlations between the linear speed on the anterior cerebral arteries and average day systolic BP. The signs of endothelial dysfunction represented by significant differences among the indices of activation of endothelium in RA patients in comparison with the control group were shown. Content of ET-1, sVCAM-1, vWF AG, Il-8, CRP was higher in RA. The number of DE was significantly higher as well. These indices showed significant differences depending on RA activity. Correlation analysis revealed that the markers of endothelium activation sVCAM-1, vWF AG were positively correlated with the indices of immune inflammation. An increase in the activity of inflammatory process in RA leads to endothelial dysfunction, which contributes to the increase in the peripheral vascular resistance of cerebral arteries, reduction in the high-speed indices of blood flow, growth of BP variability and the increase in load by pressure.