sE-selectin Research Articles

ADHESION MOLECULES IN INTESTINAL DESTRUCTIVE-INFLAMMATORY PROCESS IN THE CHILDREN WITH ULCERATIVE COLITIS

Aim : to study the content of serum soluble cell adhesion molecules in children with ulcerative colitis that mediate the initial and final stages of the migration of leukocytes to the focus of inflammation: sP-selectin (soluble platelet selectin) and Specam-1 (soluble platelet-endothelial cell adhesion molecule 1) as well some earlier unexplored factors associated with their level. Patients and methods : we examined 107 patients with ulcerative colitis aged from 6 up to 17 years. The diagnosis was set on the base of a comprehensive examination. The content of serum soluble adhesion molecules sP-selectin and sPECAM-1 as well cytokine status and neopterin were evaluated by ELISA. Respiratory metabolism was investigated by using chemiluminescent reactions. Results : it was shown that the content of sP-selectin and sPECAM-1 is significantly higher in patients than in the control group, which may influence on the migration of leukocytes into tissues for realization of their effector potential. It is confirmed by morphological analyses of the intestine biopsies, where it was observed the increasing of the number of leukocytes in vascular endothelium and epithelial layer. At the same time strengthening of the oxygen-dependent metabolism of neutrophils, the increase of the concentration of neopterin and tumor necrosis factor α were noted. Conclusions : the correlation of the studied adhesion molecules with a number of inflammatory markers (TNFα (tumor necrosis factor α), free radicals, neopterin) was revealed, which indicates the diagnostic value of serum levels of the membrane antigens. The increase of the concentration of adhesion molecules sP-selectin and sPECAM-1 may be one of the links of the pathogenesis of ulcerative colitis.

Endothelial Activation and Coagulation Homeostasis Play a Role in Coronary Allograft Vasculopathy after Heart Transplantation in Children

Purpose Inflammation and endothelial dysfunction are involved in coronary allograft vasculopathy (CAV) after heart transplant (HT) but the mechanisms and the potential impact on the native vasculature is unknown. Methods and Materials We used IVUS to evaluate CAV in children after transplant and studied native, systemic vascular structure and function with carotid intima-medial thickness (cIMT) and brachial artery flow mediated dilatation (FMD). In 54 children (28 male) age 9 to 18 years, we undertook IVUS, cIMT and FMD median 58 months (2.2 to 151) after HT. Findings were compared to 16 age, sex matched, healthy controls. Circulating markers of inflammation and endothelial activation were also measured. Results 10 patients had evidence of CAV with mean intima-media thickening > 0.5 mm analysed over 26 mm (SD 9) of the LAD coronary. This represents patients in the upper quartile. No relation to time post transplant and donor age was detected. HT patients had evidence of endothelial activation with an increase in soluble e-selectin, VCAM1 and thrombomodulin in the absence of raised inflammatory cytokines. CAV was related to low levels of thrombomodulin™, tissue factor (TF) and increased von Willebrand Factor (vWF) in peripheral blood suggesting a relationship to coagulation homeostasis. In contrast there were no abnormalities of systemic vascular structure and function. Conclusions After heart transplant there was evidence of significant CAV but not systemic vasculopathy. TF, TM and vWF were related to coronary IMT suggesting involvement in pathogenesis of CAV and novel treatment opportunities. Patient Control Median Max Min Median Max Min p value IL1B 0.6 0.001 4.2 0.001 0.001 0.9 IL8 3.5 1.6 9.0 4.3 2.0 9.1 0.06 sVCAM_1 538 244 758 442 318 676 0.03* sE selectin 15 7.6 27.7 11.4 7.5 19.6 0.005* Thrombomodulin 7.0 4.4 11.3 5.2 4.4 9.1 hsCRP 0.3 0.01 8.7 0.2 0.05 3.4 0.9 IL6 3.9 1.4 20.7 3.1 1.7 13.3 0.3 TNF alpha 5.0 2.7 10.9 5.3 3.6 8.0 0.3 Mean IMT ng/ml > 0.5 mm p value Thrombomodulin 6.0 7.2 0.046 TF 27.8 35.3 0.023 vWF 0.79 0.54 0.013

Abstract MP67: The Association Between Habitual Dietary Sodium and Soluble Cell Adhesion Molecules Among Middle-Aged Male Twins

Introduction: Habitual dietary sodium intake is associated with coronary flow reserve, a measure of microvascular function. We investigated the role of habitual dietary sodium in the expression of soluble cell adhesion molecules; these molecules are markers of endothelial dysfunction that are thought to contribute to the development of atherosclerosis. Hypothesis: Habitual dietary sodium is directly associated with soluble vascular cell adhesion molecule-1 (sVCAM-1) and soluble intercellular adhesion molecule-1 (sICAM-1) concentration. Methods: We recruited 475 predominantly healthy middle-aged male twins, including 214 monozygotic and dizygotic twin pairs and 47 unpaired twins, from the Vietnam Era Twin Registry. The twins had no previous history of coronary heart disease. Food intake was assessed using the Willett food-frequency questionnaire. We estimated habitual daily sodium consumption over the previous year from reported food intake. We measured sVCAM-1 and sICAM-1 concentration in blood using commercially available ELISA kits. We also obtained information on demographic characteristics and traditional CVD risk factors, including blood pressure. Mixed-effect regression analysis was used to examine individual-level effects and robust regression analysis was used to examine within-pair differences. Results: The twins’ mean age was 53.4 years (SD = 3.1), and 96% (455 out of 475) were white. An increase in dietary sodium of 1,000 mg/d was associated with a 12.2% higher sVCAM-1 concentration (95% CI: 3.1, 21.2; P = 0.009) after adjustment for total energy intake, various nutritional factors, demographic characteristics, and traditional CVD risk factors. For sICAM-1, the adjusted individual-level effect was only 3.1% and did not achieve statistical significance ( P = 0.39). Across quintiles of sodium consumption, sVCAM-1 concentration was directly associated with dietary sodium ( P for trend = 0.002) with the top quintile (sodium >1,505 mg/d) having a 22.9% higher sVCAM-1 concentration than the bottom quintile (sodium <742 mg/d) after controlling for potential confounders. This association persisted within twin pairs ( P = 0.03), yet differed by zygosity ( P interaction = 0.01). Among dizygotic pairs, a 1,000 mg/d within-pair difference in dietary sodium was associated with a 17.3% higher sVCAM-1 concentration in the twin with higher dietary sodium than in the co-twin with lower dietary sodium ( P = 0.005) after adjustment for potential confounders. Among monozygotic pairs, however, the within-pair difference was only 5.0% per 1,000 mg/d of sodium intake and did not reach statistical significance ( P = 0.25). Conclusions: Habitual dietary sodium is directly associated with sVCAM-1 concentration, a marker of abnormal endothelial function, independent of traditional CVD risk factors. However, shared genetic factors may mediate this association.

ATP-Binding Cassette Transporter B4 Anchors the Cell Adhesion Molecule DdCAD-1 to Cell Membrane in Dictyostelium discoideum

In Dictyostelium, soluble cell adhesion molecule, DdCAD-1, regulates cell-cell interaction through an unknown anchoring protein on the plasma membrane. Far western blot analysis using different probes revealed that the potential DdCAD-1 interacting protein was between 64 and 98kDa. To isolate and identify the anchoring protein, GST-DdCAD-1 and anchoring protein were cross-linked in vivo by chemical cross-linker and stable protein complex was isolated by co-immunoprecipitation assays. The protein cross-linked to DdCAD-1 was extracted from the gel slice and trypsinized. The peptides were subjected to analysis by mass spectrometry, which showed that the putative anchoring protein belongs to ATP-binding cassette transporter family.

Hypolipidaemic and anti-inflammatory effects of fixed dose combination of atorvastatin plus ezetimibe in Indian patients with dyslipidaemia

We aimed to assess the efficacy of fixed dose combination of atorvastatin plus ezetimibe in Indian patients with dyslipidaemia. A double-blind study was conducted to assess the effect of fixed dose combination of ezetimibe 10 mg plus atorvastatin 10 mg on lipid profile, oxidised low-density lipoprotein (ox-LDL), high-sensitivity C-reactive protein (hsCRP) and soluble intercellular cell adhesion molecule (sICAM) in dyslipidaemic patients with or at high risk of coronary artery disease, and compare it with atorvastatin 10 mg monotherapy. 30 patients were randomised to receive ezetimibe plus atorvastatin or atorvastatin once daily for four weeks. Of the 30 patients, 10 men and 5 women (mean age 54.3 ± 1.6 years) received ezetimibe plus atorvastatin, while 13 men and 2 women (mean age 53.7 ± 2.8 years) received only atorvastatin. The combination treatment significantly reduced total cholesterol (percentage treatment difference -14.4 ± 6.5, 95% confidence interval [CI] -1.0 to -27.7; p = 0.041) and LDL cholesterol (LDL-C; percentage treatment difference -19.9 ± 6.1, 95% CI -7.4 to -32.4; p = 0.003) compared to atorvastatin monotherapy. 13 patients on combination treament achieved the National Cholesterol Education Program target for LDL-C as compared to 9 patients on atorvastatin monotherapy (p = 0.032). Significant reductions in very low-density lipoprotein cholesterol, triglyceride, ox-LDL and sICAM were observed with combination treatment compared to atorvastatin monotherapy. However, no significant change was seen in high-density lipoprotein cholesterol or hsCRP levels between the two groups. Combination treatment with atorvastatin and ezetimibe had relatively better lipid-lowering and anti-inflammatory efficacy than atorvastatin monotherapy.

The Effects of Arterial Blood Pressure Reduction on Endocan and Soluble Endothelial Cell Adhesion Molecules (CAMs) and CAMs Ligands Expression in Hypertensive Patients on Ca-Channel Blocker Therapy

Background/Aims: To determine the effect of arterial blood pressure (BP) reduction on endocan and soluble cell adhesion molecules' (sCAM) plasma concentration and expression of their ligands on circulatory leukocyte subpopulations. Methods: 24 hypertensive subjects of both sexes (age: 53±8 yrs) were treated with Ca-channel blocker, amlodipin (5-10 mg/day for 8 weeks; to reach BP≤139/89mmHg). The serum sCAMs and endocan concentrations were determined by ELISA kits. Level of ICAM/VCAM ligands on leukocytes was assessed by flow cytometry. Paired t-test, or t-test were used as appropriate, with Pearson's correlation calculated; p<0.05 was considered significant (SigmaPlot v.11). Results: sICAM-1 and sVCAM-1 were decreased (p≤0.001 and p=0.002, respectively), while E-selectin concentration was increased after amlodipin treatment (P=0.014). CD11a/LFA-1 (ICAM-1 and endocan ligand) was significantly increased in all three cell types with BP decrease. CD15 and CD49d/VLA-4 (VCAM-1 ligand) did not change after the treatment. There was significant positive correlation of systolic and diastolic BP with ICAM-1 and VCAM-1, and significant negative correlation of systolic BP with CD11a/LFA-1. Endocan significantly positively correlated with ICAM-1. Conclusions: The increased expression of ICAM/VACM ligands, together with decrease of sCAMs and endocan suggests the de-activation of endothelium with reduction in BP, decreasing the adherence of circulatory leukocytes to endothelium; subsequently decreasing the risk for development of atherosclerosis.

Increased expression of cell adhesion molecule receptors on monocyte subsets in ischaemic heart failure

The objective of this study was to evaluate the expression of cell adhesion molecule (CAM) receptors (integrins) on monocyte subsets in heart failure (HF) and examine their prognostic implication.Increased levels of soluble CAMs have been observed in patients with HF, but the precise mechanism of monocyte adhesion to the vascular endothelium remains unknown. Patients with acute HF (AHF, n=51) were compared to those with stable HF (SHF, n=42) and stable coronary artery disease (CAD, n=44) without HF. Expression of integrins-receptors to intercellular adhesion molecule-1 (ICAM-1R) and vascular CAM-1 (VCAM-1R) on monocyte subsets was assessed by flow cytometry. Monocyte subsets were defined as CD14++CD16-CCR2+ ('classical', Mon1), CD14++CD16+CCR2+ ('intermediate', Mon2), and CD14+CD16++CCR2- ('non-classical', Mon3). Compared to patients with SHF, those with AHF had significantly higher expression of ICAM-1R on Mon2 (p=0.01). Compared to those with stable CAD, patients with SHF had a significantly higher expression of ICAM-1R on Mon2 (p=0.025).Compared to SHF, patients with AHF had a similar expression of VCAM-1R on both Mon1 and Mon3 but significantly higher expression on Mon2 (p=0.019). There were no significant differences between SHF and CAD in monocyte expression of VCAM-1R. In multivariate Cox regression analysis, VCAM-1R expression on Mon2 was associated with adverse clinical outcome (death or rehospitalisation) in AHF [HR 1.07 (1.01-1.14), p=0.029]. In conclusion, HF is associated with increased monocyte expression of integrins-receptors to both ICAM-1 and VCAM-1, being particularly linked to Mon2 subset. Expression of VCAM-1R on Mon2 may have prognostic value in patients with AHF.

Improved Arteriole‐to‐Venule Ratio of Retinal Vessels Resulting From Bariatric Surgery

Obesity causes increased morbidity and mortality from metabolic and cardiovascular disease (CVD). We investigated the effect of bariatric surgery on endothelial dysfunction (ED) in retinal vessels as a marker of metabolic and cardiovascular risk in patients with obesity WHO III.Thirty consecutive patients (19/11, w/m) were evaluated by anthropometry, lipid profile, and oral glucose tolerance test before and after bariatric surgery (Mannheim Obesity Study (MOS); NCT 00770276). Risk stratification was performed by the presence of metabolic syndrome (MetS) according to ATP-III (adult treatment panel-III). Subclinical atherosclerosis was assessed by measurement of intima-media thickness (IMT). Flicker light response of retinal vessels was used as measures of ED. We measured their arteriole-to-venule ratio (AVR) for evaluation of vascular pathology. After a median of 9 months following bariatric surgery, mean weight loss was 39.4 kg (37.3%). Remission of impaired glucose metabolism was achieved in 53.3% of affected patients. Dyslipidemia improved significantly (triglycerides -61.3 mg/dl, P < 0.0001, total cholesterol -28.2 mg/dl, P = 0.002, and low-density lipoprotein cholesterol were reduced -24.5 mg/dl, P = 0.008). This resulted in a significant reduction of patients classified for MetS (27 vs. 9, P < 0.0001). Adiponectin increased by 2.08 µg/l (P = 0.032) and high sensitivity C-reactive protein (hs-CRP) and soluble intercellular cell adhesion molecule (sICAM) decreased (-7.3 mg/l, P < 0.0001 and -146.4 ng/ml, P = 0.0006). AVR improved significantly (+0.04, P < 0.0001), but neither Flicker light response nor IMT changed significantly. Retinal AVR is ameliorated after bariatric intervention. As an increased AVR results from either or both widening retinal arteriolar caliber and narrowing retinal venular caliber, an improvement in small vessel profile is evident 9 months after bariatric surgery.

Association of adipokines and adhesion molecules with indicators of obesity in women undergoing mammography screening

BackgroundThe soluble cell adhesion molecules and adipokines are elevated in patients with obesity, hypertension, type 2 diabetes mellitus, breast cancer and atherosclerosis.ObjectiveTo investigate the relationship between anthropometric profile, dietary intake, lipid profile and fasting glycemia with serum levels of adipokines (adiponectin and PAI-1) and adhesion molecules (ICAM-1 and VCAM-1) in women without breast cancer undergoing routine mammographic screening.DesignTransversal study.SubjectsOne hundred and forty-five women over 40-years old participated in this study.ResultsIn 39.3% of cases the BMI was above 30 kg/m2; 46.9% had hypertension, 14.5% had type 2 Diabetes Mellitus, 31.7% had dyslipidemia and 88.3% presented a waist-to-hip ratio ≥ 0.8. A linear correlation was found between serum levels of PAI-1 and triglycerides, between serum levels of PAI-1 and WHR and between serum levels of VCAM-1 and BMI.ConclusionWe found a high prevalence of obesity and metabolic syndrome. PAI-1 and VCAM-1 levels were correlated with clinical indicators of obesity and overweight.

MMPs and Soluble ICAM-5 Increase Neuronal Excitability within In Vitro Networks of Hippocampal Neurons

Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases that are released from neurons in an activity dependent manner. Published studies suggest their activity is important to varied forms of learning and memory. At least one MMP can stimulate an increase in the size of dendritic spines, structures which represent the post synaptic component for a large number of glutamatergic synapses. This change may be associated with increased synaptic glutamate receptor incorporation, and an increased amplitude and/or frequency of α-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate (AMPA) mini excitatory post-synaptic currents (EPSCs). An associated increase in the probability of action potential occurrence would be expected. While the mechanism(s) by which MMPs may influence synaptic structure and function are not completely understood, MMP dependent shedding of specific cell adhesion molecules (CAMs) could play an important role. CAMs are ideally positioned to be cleaved by synaptically released MMPs, and shed N terminal domains could potentially interact with previously unengaged integrins to stimulate dendritic actin polymerization with spine expansion. In the present study, we have used multielectrode arrays (MEAs) to investigate MMP and soluble CAM dependent changes in neuronal activity recorded from hippocampal cultures. We have focused on intercellular adhesion molecule-5 (ICAM-5) in particular, as this CAM is expressed on glutamatergic dendrites and shed in an MMP dependent manner. We show that chemical long-term potentiation (cLTP) evoked changes in recorded activity, and the dynamics of action potential bursts in particular, are altered by MMP inhibition. A blocking antibody to β1 integrins has a similar effect. We also show that the ectodomain of ICAM-5 can stimulate β1 integrin dependent increases in spike counts and burst number. These results support a growing body of literature suggesting that MMPs have important effects on neuronal excitability. They also support the possibility that MMP dependent shedding of specific synaptic CAMs can contribute to these effects.

Effect of minocycline postconditioning and ischemic postconditioning on myocardial ischemia-reperfusion injury in atherosclerosis rabbits

This study examined the protective effect of ischemic postconditioning (IPoC) and minocycline postconditioning (MT) on myocardial ischemia-reperfusion (I/R) injury in atherosclerosis (AS) animals and the possible mechanism. Forty male healthy rabbits were injected with bovine serum albumin following feeding on a high fat diet for 6 weeks to establish AS model. AS rabbits were randomly divided into 3 groups: (1) I/R group, the rabbits were subjected to myocardial ischemia for 35 min and then reperfusion for 12 h; (2) IPoC group, the myocardial ischemia lasted for 35 min, and then reperfusion for 20 s and ischemia for 20 s [a total of 3 cycles (R20s/I20s×3)], and then reperfusion was sustained for 12 h; (3) MT group, minocycline was intravenously injected 10 min before reperfusion. The blood lipids, malondialdehyde (MDA), superoxide dismutase (SOD), soluble cell adhesion molecule (sICAM), myeloperoxidase (MPO), and cardiac troponin T (cTnT) were biochemically determined. The myocardial infarction size (IS) and apoptosis index (AI) were measured by pathological examination. The expression of bcl-2 and caspase-3 was detected in the myocardial tissue by using reverse transcription-polymerase chain reaction (RT-PCR). The results showed that the AS models were successfully established. The myocardial IS, the plasma levels of MDA, sICAM, MPO and cTnT, and the enzymatic activity of MPO were significantly decreased, and the plasma SOD activity was significantly increased in IPoC group and MT group as compared with I/R group (P<0.05 for all). The myocardial AI and the caspase-3 mRNA expression were lower and the bcl-2 mRNA expression was higher in IPoC and MT groups than those in I/R group (all P<0.05). It is concluded that the IPoC and MT can effectively reduce the I/R injury in the AS rabbits, and the mechanisms involved anti-oxidation, anti-inflammation, up-regulation of bcl-2 expression and down-regulation of caspase-3 expression. Minocycline can be used as an effective pharmacologic postconditioning drug to protect myocardia from I/R injury.

Serum Levels of Soluble Adhesion Molecules as Prognostic Factors for Acute Liver Failure

Background/Aims: In patients with septic shock, the degree of liver dysfunction is correlated with serum levels of soluble intercellular adhesion molecule (sICAM)-1. We aimed to assess the usefulness of serum levels of soluble adhesion molecules as prognostic factors for acute liver failure (ALF). Methods: Serum levels of soluble platelet endothelial cell adhesion molecule (sPECAM)-1, sICAM-3, soluble endothelial (sE) selectin, sICAM-1, soluble platelet selectin, and soluble vascular cell adhesion molecule-1 on admission were measured in 37 ALF patients and 34 healthy controls. Results: Twenty-two ALF patients (59%) reached to fatal outcomes. Serum levels of sPECAM-1, sICAM-3, sE-selectin and sICAM-1 were higher in ALF patients than healthy controls. In 37 ALF patients, by the multivariate logistic regression analysis, ratio of direct to total bilirubin (per 0.1 increase; OR 0.11, 95% CI 0.01–0.99), serum sPECAM-1 level (per 100 ng/ml increase; OR 4.37, 95% CI 1.23–15.5) and serum sICAM-1 level (per 100 ng/ml increase; OR 0.49, 95% CI 0.27–0.89) were associated with fatal outcomes. Using receiver operating characteristics curve, each area under the curve of serum sPECAM-1 and sICMA-1 levels as prognostic factors was 0.71 and 0.74, respectively. Conclusion: Serum sPECAM-1 and sICAM-1 levels may be useful for predicting the prognosis of ALF.

Changes of serum IL-18, VEGF, and sVCAM-1 in patients with primary hepatic carcinoma and their clinical significance

Objective To investigate serum levels of interleukin-18 ( IL-18 ), vascular endothelial growth factor ( VEGF ),and soluble vascular cell adhesion molecule-1 ( sVCAM-1 ) in patients with primary hepatic carcinoma ( PHC ) and their clinical significance.Methods Serum levels of IL-18,VEGF,and sVCAM-1 were detected in 15 patients with PHC by enzyme-linked immunosorbent assay ( ELISA ) before and after treatment.The findings were compared with those from the healthy controls.Results The baseline levels of IL-18,VEGF,and sVCAM-1 were significantly higher in the patients with PHC than in the healthy controls ( P＜ 0.05 ).Levels of IL-18,VEGF,and sVCAM-1 were decreased two weeks after treatment as compared with the baselines ( P＜ 0.05 ),but were still higher than those from the healthy controls ( P ＜ 0.05 ).Conclusions An elevation in serum levels of IL-18,VEGF,and sVCAM-1 in patients with primary hepatic carcinoma may be associated with the genesis,development,and invasion of the malignancy. Key words: [Key words] Hepatic carcinoma; Interleukin-18; Vascular endothelial growth factor; Soluble vascular cell adhesion molecule- 1

The Effect of Soy Protein Beverages on Serum Cell Adhesion Molecule Concentrations in Prehypertensive/Stage 1 Hypertensive Individuals

Objective: Prehypertensive and hypertensive individuals are at increased risk of atherosclerotic cardiovascular disease (CVD), in part because hypertension contributes to endothelial dysfunction and increased cell adhesion molecule expression. Soy protein and isoflavones may favorably alter CVD risk factors, and hence the aim of this study was to determine whether intake of cow's milk compared with soy beverage prepared from whole soy bean (WSB) or soy protein isolate (SPI) would lower soluble cell adhesion molecule concentrations as a means of decreasing CVD risk.Methods: We enrolled healthy prehypertensive/stage 1 hypertensive men (n = 60; 18–63 years) and premenopausal women (n = 8; 20–48 years). Participants were randomized to 1 of 3 groups for 8 weeks: cow's milk (600 mL/d), SPI beverage (840 mL/d; 30.1 mg total isoflavones/d), or WSB beverage (840 mL/d; 91.4 mg total isoflavones/d). We measured soluble vascular cell adhesion molecule-1 (VCAM-1), intercellular cell adhesion molecule-1 (ICAM-1), and endothelial-leukocyte adhesion molecule-1 (E-selectin) concentrations at baseline and week 8.Results: Soluble CAM concentrations were not altered by treatment and did not differ between prehypertensive and hypertensive participants. However, analysis of variance indicated a treatment × gender interaction (gender effect) for ICAM-1 (p = 0.0037) but not for E-selectin (p = 0.067) or VCAM-1 (p = 0.16). Men had higher concentrations of ICAM-1 and E-selectin, respectively, at baseline (p = 0.0071, p = 0.049) and week 8 (p = 0.0054, p = 0.038) than women did.Conclusion: Neither intake of cow's milk nor soy beverage for 8 weeks altered soluble CAM concentrations in prehypertensive/stage 1 hypertensive individuals, suggesting that neither type of beverage diminished atherosclerotic CVD risk in mildly hypertensive individuals by way of improving circulating CAM concentrations.

Impact of intestinal ischemia/reperfusion and lymph drainage on distant organs in rats

To investigate the impact of intestinal ischemia/reperfusion (I/R) injury and lymph drainage on distant organs in rats. Thirty-two Sprague-Dawley male rats, weighing 280-320 g, were randomly divided into blank, sham, I/R, and ischemia/reperfusion and drainage (I/R + D) groups (n = 8). All rats were subjected to 60 min ischemia by clamping the superior mesenteric artery, followed by 120 min reperfusion. The rats in the I/R + D group received intestinal lymph drainage for 180 min. In the sham group, the abdominal cavity was opened for 180 min, but the rats received no treatment. The blank group served as a normal and untreated control. A chromogenic limulus assay kit was used for quantitative detection of serum endotoxin. The serum concentrations of tumor necrosis factor-α (TNF-α), interleukin (IL)-6, IL-1β, soluble cell adhesion molecules (sICAM-1), and high mobility group protein box 1 (HMGB1) were determined with an enzyme-linked immunosorbent assay kit. Histological evaluations of the intestine, liver, kidney, and lung were performed by hematoxylin and eosin staining and immunohistochemistry. HMGB1 protein expression was assayed by western blot analysis. The serum levels of endotoxin and HMGB1 in the I/R and I/R + D groups were significantly higher than those in the sham group (endotoxin, I/R and I/R + D vs sham: 0.033 ± 0.004 EU/mL, 0.024 ± 0.003 EU/mL vs 0.017 ± 0.009 EU/mL, respectively, P < 0.05; HMGB1, I/R and I/R + D vs sham: 5.473 ± 0.963 EU/mL, 4.906 ± 0.552 EU/mL vs 0.476 ± 0.406 EU/mL, respectively, P < 0.05). In addition, endotoxin and HMGB1 were significantly lower in the I/R + D group compared to the I/R group (P < 0.05). The serum inflammatory factors IL-6, IL-1β, and sICAM-1 in the I/R and I/R + D groups were significantly higher than those in the sham group (IL-6, I/R and I/R + D vs sham: 41.773 ± 9.753 pg/mL, 19.204 ± 4.136 pg/mL vs 11.566 ± 2.973 pg/mL, respectively, P < 0.05; IL-1β, I/R and I/R + D vs sham: 144.646 ± 29.378 pg/mL, 65.829 ± 10.888 pg/mL vs 38.178 ± 7.157 pg/mL, respectively, P < 0.05; sICAM-1, I/R and I/R + D vs sham: 97.360 ± 12.714 ng/mL, 48.401 ± 6.547 ng/mL vs 33.073 ± 5.957 ng/mL, respectively; P < 0.05). The serum TNF-α in the I/R group were significantly higher than in the sham group (45.863 ± 11.553 pg/mL vs 18.863 ± 6.679 pg/mL, respectively, P < 0.05). These factors were significantly lower in the I/R + D group compared to the I/R group (P < 0.05). The HMGB1 immunohistochemical staining results showed no staining or apparent injury in the blank group, and slight staining at the top of the microvillus was detected in the sham group. In the I/R group, both the top of villi and the basement membrane were stained for HMGB1 in most areas, and injury in the I/R + D group was less than that in the I/R group. HMGB1 expression in the liver, kidney, and lung of rats in the I/R + D group was significantly lower than the rats in the I/R group (P < 0.05). Lymph drainage could block the "gut-lymph" pathway, improve intestinal barrier function, and attenuate distant organ injury incurred by intestinal I/R.

Serum levels of sICAM-1, sVCAM-1, sE-selectin, sP-selection in healthy Bulgarian people

Endothelial dysfunction is increasingly recognized as an important early feature of vascular disease. As the damage to endothelium is a key underlying factor in the development and progression of atherosclerotic processes, markers of endothelial abnormalities have been sought. Increased expression of cell adhesion molecules (CAMs) on the vascular endothelium has been postulated to play a significant role in atherogenesis. Both in vitro and in vivo studies have suggested that different risk factors of atherosclerosis may increase expression of CAMs. The elevated level of soluble forms of CAMs in circulation is associated with a higher risk to future cardiovascular events in subjects predisposed to atherosclerosis To determine the reference range for serum concentration of soluble cell adhesion molecules - slCAM-1, sVCAM-1, sE-selectin, sP-selectin. We studied 110 healthy people of Bulgarian nationality aged 18-65. The selection criteria for the reference group were made in accordance with the requirements of the International Federation of Clinical Chemistry (IFCC). Serum concentrations of CAMs were analysed by means of ELISA assay. The results are presented as central 95% interval and 0.90 confidence interval of the reference range. Reference ranges were determined for sICAM-1 (128.9 - 347.48 ng/ml), sVCAM-1 (170.42 - 478.36 ng/ml), sE-selectin (9.15 - 65.19 ng/ml) and sP-selectin (101.86 - 209.7 ng/ml). As we found no sex-related differences in the CAMs concentrations (p > 0.05) there needed to be no separate reference intervals for men and women. The single-factor dispersion analysis we used in analysing the effect of age found no age-related dependence (p > 0.05, F = 1.038) for the serum CAM concentrations in the 18-65 age range, which means that it is not necessary to establish reference intervals for smaller age ranges in this age group. The reference ranges for slCAM-1, sVCAM-1, sE-selectin, sP-selectin computed in accordance with the results distribution can be used as baseline criteria in clinical laboratory studies.

Delayed asthmatic response: a new phenotype of bronchial response to allergen challenge and soluble adhesion molecules in the serum

Patients with bronchial asthma develop various types of asthmatic response to bronchial challenge with allergen, such as immediate asthmatic response, late asthmatic response, or delayed asthmatic response (DYAR), due to different immunologic mechanisms. To investigate the appearance and possible changes in the concentrations of soluble cell adhesion molecules during the DYAR, to explore the involvement of particular cell types in the mechanism(s) leading to DYAR, and to contribute to a fuller understanding of this clinical phenomenon. The DYAR recorded in 28 patients (P < .001), appearing within 26 to 32 hours, reaching maximum within 32 to 48 hours, and resolving within 56 hours after the allergen challenge, was repeated 2 to 6 weeks later. The repeated DYAR (P < .001) was supplemented with blood cell counts and measurement of serum concentrations of soluble adhesion molecules by an enzyme-linked immunoassay. The prechallenge concentrations of soluble intercellular adhesion molecule 1 (sICAM-1), soluble vascular cell adhesion molecule 1 (sVCAM-1), soluble platelet endothelial cell adhesion molecule (sPECAM-1), soluble E-selectin, soluble L-selectin, soluble P-selectin, and soluble E-cadherin did not differ significantly from healthy controls. The DYAR was associated with the following changes in the serum: an increase of sICAM-1 at 6 and 12 hours and a decrease at 24 hours; an increase of sVCAM-1 at 24 and 36 hours; an increase of sPECAM-1 at 36 and 48 hours and a decrease at 56 and 72 hours; an increase of soluble E-selectin at 56 hours; an increase of soluble L-selectin at 56 and 72 hours; a decrease of soluble E-cadherin at 48 and 56 hours; and increased counts of blood leukocytes at 36, 48, and 56 hours, neutrophils at 24, 36, 48, and 56 hours, lymphocytes at 24, 36, and 48 hours, and monocytes at 6, 12, and 24 hours. The Th1/Th2 ratio in blood increased at 24, 36, 48, and 56 hours. The intracellular concentration of interferon γ, but not of interleukin 4, increased at 24, 36, 48, and 56 hours. These results provide evidence of the involvement of neutrophils, Th1 lymphocytes, monocytes, platelets, and endothelial cells, upon participation of various adhesion molecules, in mechanisms(s) underlying the clinical DYAR.

Relevance of erythrocyte deformability to the concentration of soluble cell adhesion molecules and glomerular filtration rate in patients with untreated essential hypertension

Relationship between erythrocyte deformability and: a) soluble cell adhesion molecules concentration, b) glomerular filtration rate (eGFR) has been investigated in three study groups: a group of 20 patients with diagnosed arterial hypertension, a group of 20 individuals with exclusively hypercholesterolemia and a group of 22 healthy persons. The individuals with hypertension or hypercholesterolemia were free of any other cardiovascular disease risk factor and were not on any therapy prior to entering the study. Clinical and laboratory data included systolic and diastolic blood pressure (obtained by ABPM), lipids profile, eGFR, red blood cell (RBC) deformability (assessed by shear stress laser diffractometry) and levels of circulating soluble vascular adhesion molecules-1 (sVCAM-1) as well as soluble intracellular adhesion molecules-1 (sICAM-1). In the group of hypertensives, RBC deformability and concentration of circulating soluble adhesion molecules showed statistically significant negative correlations: RBC deformability decreases with increasing level of: a) sVCAM-1, R = -0.61, p < 0.002, b) sIVCAM-1, R = -0.53, p < 0.009. In parallel, statistically significant increase of eGFR was observed with rising erythrocyte deformability, R = 0.60, p < 0.005. In the groups of healthy individuals and patients with hypercholesterolemia there was no sign of any correlations between the considered parameters. The observed correlations suggest that in patients diagnosed exclusively with hypertension, firstly, erythrocyte deformability may serve as a marker of endothelial dysfunction and, secondly, red blood cells may be mediators of adverse changes in kidneys.

Detection and clinical significance of serum soluble E-selectin(sE-selectin)and serum soluble P-selectin(sp-selectin)in patients with esophageal squamous cell carcinoma

Objective To study the dinical value of serum soluble E-selectin(sE-selectin)and serum soluble P-selectin(sp-selectin)in patients with esophageal squamous cell carcinoma. Methods The sP-selectin and sP-selectin of the serum was measured by ELISA in 27 patients with viral encephalitis and 35 normal persons. Results In patients with organ metastasissE-selectin and sP-selectin levels were significantly higher than normal(P＜0.01),In patients with organ metastasis,sE-selectin and sP-selectin levels increased more pronounced.No visceral metastasis in patients with esophageal cancer,postoperative serum sE-selectin and sP-selectin levels significantly decreased compared with the preoperative(P＜0.01),there are organ metastasis in patients with esophageal cancer sE-selectin and sP-selectin levels decreased not obvious. Conclusion sE-selectin and sP-selectin could be used as a new tumor detection of esophageal cancer targets,and has close relatimship with the incidence of esophageal cancer,development and metastasis. Key words: Esophageal neoplasms; E-selectin; P-selectin

Relationship of Soluble Adhesion Receptors to Red Cell Apoptosis In SCD.

AbstractAbstract 1654The interaction of sickle red cells (SS RBCs) with endothelium can cause endothelial injury, leading to increased expression of soluble endothelial cell adhesion molecules and plasma markers of endothelial activation and dysfunction. Among such plasma markers, sVCAM-1 levels are increased in SCD and rise further during vaso-occlusive crisis (Duits et al. 1993), and several groups have shown that sVCAM-1 levels are positively related to LDH levels, a marker of hemolysis. Perfusion of endothelial cells with SS RBCs has been shown to increase the expression of the cellular adhesion molecules ICAM-1 and VCAM-1 by endothelial cells (Shiu et al. 2000). Furthermore, sVCAM-1, sICAM-1, and sE-selectin have been independently associated with risk of mortality (Kato et al. 2005). We were interested in determining whether identifiable characteristics of SS RBCs themselves might be associated with the degree to which soluble adhesion molecules are expressed in SCD patient plasma. Therefore, we measured sVCAM-1, sICAM-1, sCD40 ligand (sCD40L) and sP-selectin in SCD patient plasma by ELISA (n=46) and looked at the relationship of these biomarkers of disease activity to SS RBC characteristics. We specifically examined phosphatidylserine exposure (as measured by annexin V binding analyzed by flow cytometry) and the degree of SS RBC adhesion to laminin and endothelial cells, measured in vitro in a graduated height flow chamber. We found that the degree of red cell damage measured by annexin V binding correlated with the level of soluble adhesion receptors sVCAM-1 (p=0.036) and sICAM-1 (p=0.026). However, annexin V binding did not correlate with sCD40L or sP-selectin expression in plasma, and neither annexin V binding nor levels of soluble adhesion molecules correlated with SS RBC adhesion to laminin or endothelial cells in vitro. In addition, the concentrations of sVCAM-1 and sICAM-1 in plasma were strongly correlated with each other (p<0.0001), suggesting that both of these adhesion molecules may be released from activated endothelial cells in SCD patients as a result of the same mechanism involving contact with phosphatidylserine-exposing SS RBCs. These data support the hypothesis that phosphatidylserine exposure on SS RBCs is a critical factor in SS RBC interaction with the endothelium (Setty et al., 2002), including the release of soluble cellular adhesion markers that indicate endothelial activation and/or injury. Disclosures:Telen:GlycoMimetics, Inc.: Consultancy.