sVCAM-1 Levels Research Articles

Enhanced susceptibility to alcohol-induced bacterial translocation, immune activation, and persistent inflammation in patients with alcoholic hepatitis: a prospective observational study

Abstract Up to 30% chronic heavy drinkers develop a spectrum of severe alcoholic liver diseases, including alcoholic hepatitis (AH). Alcohol-induced impairment of intestinal integrity and microbial translocation into the portal vein lead to immune activation and play a major role in the pathogenesis of AH. To better understand why AH develops only in some heavy drinkers and the effect of alcohol abstinence on immunity, we conducted a prospective study to compare plasma LPS levels and peripheral blood immunoprofiles between 68 AH patients and 65 heavy drinkers without liver disease (HDC) who were followed up to 12 months. Plasma samples were also obtained from 31 health donors (HD). Plasma LPS was detected in a higher portion of AH patients than HDC (80% vs 49%) at baseline, and in only 8% HD. However, LPS levels were not different between AH an HDC subjects with detectable LPS. Compared with HDC, AH patients had higher baseline plasma levels of sCD14, sCD163, sICAM-1, sVCAM-1, IL-22, TNF-α, IL-8, IP10, IL-4, IL-6, IL-9, IL-10, FGF2, IL-7, IL-15, and TGF-α, but lower levels of the anti-inflammatory chemokine MDC. AH patients also had more activated, yet dysfunctional monocytes, T cells, and B cells as they expressed higher levels of CD38 and CD69, but lower levels of HLA-DR, CD80, and CD86. In spite of having significant bacterial translocation, HDC showed little evidence of immune activation. With alcohol abstinence, levels of many dysregulated immune markers normalized in AH patients. However, TNF-α, IL-8, IL-10, sCD163, and sVCAM-1 levels remained higher. Conclusion AH patients were more susceptible to alcohol-induced bacterial translocation and immune activation. Alcohol abstinence reduced but did not fully reverse immunological abnormalities.

Abstract A26: Body fatness and adipose tissue subtypes are associated with circulating biomarkers of inflammation and angiogenesis in colorectal cancer patients: The ColoCare Study

Abstract Background: Adiposity has been linked to both risk and prognosis of colorectal cancer; however, the impact of different compartments of adipose tissue (visceral vs. subcutaneous) is unclear. In healthy individuals, adiposity is associated with elevated biomarkers of inflammation, which provides a mechanistic link between adiposity and cancer risk. For prognosis, the downstream effects of inflammation on angiogenesis may be central. We investigated associations between adiposity and biomarkers of inflammation, as well as angiogenesis, in colorectal cancer patients enrolled in the ColoCare Study, an international multicenter patient cohort. Methods: We utilized preoperatively obtained serum samples of patients with newly diagnosed colorectal cancer [n=164; (stage I-IV)] from the ColoCare Study in Heidelberg, Germany, with available diagnostic multi-detector-CT images for adipose tissue quantification. Abdominal adipose tissue was assessed by area-based quantification of visceral (VAT), and subcutaneous adipose tissue (SAT), as well as their ratio (VAT/SAT) on levels L3/L4 and L4/L5. Body mass index (BMI) was calculated (kg/m2) and demographic and clinical-surgical data were abstracted from medical records. Circulating CRP, SAA, VEGF-A, VEGF-D, sICAM-1 and sVCAM-1 levels were assessed on the Meso Scale Discoveries platform with V-plex assays at the Huntsman Cancer Institute (average intra-plate CVs 2.9%, inter-plate CVs 7.9%). Partial correlations and regression analyses, adjusting for age, sex and tumor stage were performed. Results: While overall obesity (BMI) was only significantly associated with sVCAM (r=0.23, p=0.006), visceral adiposity (VAT) was associated with both CRP and SAA (r=0.21, p=0.01 and r=0.17, p=0.04, respectively). There was no association between SAT and the measured biomarkers. Most predictive was the ratio of VAT/SAT on level L3/L4, which was associated with CRP (r=0.18, p=0.04), SAA (r=0.24, p=0.006), sICAM-1 (r=0.18, p=0.04), and particularly VEGF-A (r=0.28, p=0.0008). Similar associations were observed for the VAT/SAT ratio on level L4/5. Conclusions: We demonstrated a link between specifically visceral adiposity and biomarkers of inflammation in colorectal cancer patients. In addition, we showed that visceral adiposity also affects circulating VEGF-A levels. This protein has various effects, including the induction of angiogenesis, vasculogenesis and endothelial cell growth, as well as the promotion of cell migration, and the inhibition of apoptosis. Our findings support a mechanistic role of visceral adipose tissue in colorectal cancer risk and potentially prognosis. Citation Format: Cornelia M. Ulrich, Jürgen Böhm, Christy Warby, Tengda Lin, Mariam Salou, Biljana Gigic, Dominique Scherer, Johanna Nattenmueller, Jennifer Ose, Lin Zielske, Petra Schrotz-King, Torsten Kölsch, Erin Siegel, Christopher Li, Alexis Ulrich, Hanno Glimm, Jewel Samadder, Stephen Hursting, Hans-Ulrich Kauczor. Body fatness and adipose tissue subtypes are associated with circulating biomarkers of inflammation and angiogenesis in colorectal cancer patients: The ColoCare Study. [abstract]. In: Proceedings of the AACR Special Conference: Improving Cancer Risk Prediction for Prevention and Early Detection; Nov 16-19, 2016; Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2017;26(5 Suppl):Abstract nr A26.

Acute effects of post-absorptive and postprandial moderate exercise on markers of inflammation in hyperglycemic individuals.

Systemic inflammation is involved in the development of several diseases, including cardiovascular disease and type 2 diabetes. It is known that vigorous exercise affects systemic inflammation, but less is known about exercise at lower intensities. Hyperglycemia can also entail pro-inflammatory responses; however, postprandial hyperglycemia is blunted if the meal is followed by exercise. Hypotheses were: (1) moderate physical exercise acutely affects levels of C-reactive protein (CRP) and serum soluble vascular cell adhesion molecule 1 (sVCAM-1) in hyperglycemic individuals and (2) the effect depends on whether the activity is performed in a post-absorptive or postprandial state. Twelve participants diagnosed with hyperglycemia, but not using anti-diabetic medication, underwent three test days in a randomized cross-over study; 1 control day without exercise, 1 day with 60min of treadmill walking ending 30min before breakfast, and 1 day with an identical bout of activity 30min after the start of breakfast. Food intake was strictly standardized and venous blood for CRP, and sVCAM-1 analysis was sampled at standardized timepoints during the first 3.5h after breakfast and once 24h later. Merged data from the two exercise days showed that sVCAM-1 increased from baseline (4 ± 16ng/mL) compared to the control condition (-28 ± 47ng/mL, ES = 0.7, p = 0.024). There was no statistically significant difference in changes in sVCAM-1 levels between the two exercise test days. Exercise did not affect CRP values. Moderate exercise increases sVCAM-1 in hyperglycemic individuals, whereas it does not affect CRP.

Soluble VCAM-1/soluble ICAM-1 ratio is a promising biomarker for diagnosing endometriosis.

Do cell adhesion molecules play a role in endometriosis, and can they be used as a biomarker for diagnosing endometriosis? Altered expression of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) in the endometrium and peritoneum may play a key role in endometriosis and the soluble VCAM-1/soluble ICAM-1 ratio is a promising biomarker. Cell adhesion molecules are cell surface proteins that mediate cellular adherence, inflammatory and immune responses, and cancer-related biological processes. Altered expression of VCAM-1 and ICAM-1 in women with endometriosis has been investigated previously; however, gene expression levels in tissues and protein levels in the serum have not been investigated in the same patients. We performed a prospective, longitudinal study (the Endometriosis Marker Austria) in patients who underwent a laparoscopy for benign gynecological pathology in a university-based tertiary referral center for endometriosis. From a total of 138 women who were included in the study from July 2013 through September 2014, 97 had not received hormonal treatment for at least 3 months prior to recruitment and were included in the analysis; 49 (50.5%) of these women had endometriosis, and the 48 (49.5%) who did not have endometriosis served as a control group. During laparoscopy, tissue samples were obtained from ectopic and eutopic endometrium, and from normal pelvic peritoneum. In addition, serum samples were collected immediately before and 6-10 weeks after surgery. The mRNA levels of VCAM-1, ICAM-1 and epithelial cell adhesion molecule (EpCAM) were measured using quantitative real-time PCR, and serum protein levels of soluble VCAM-1 (sVCAM-1), ICAM-1 (sICAM-1) and EpCAM (sEpCAM) were measured using ELISA and correlated with endometriosis status. The mRNA levels of both VCAM-1 and ICAM-1 were higher in ectopic endometriotic lesions than in eutopic endometrium (P < 0.001). Moreover, the mRNA levels of both VCAM-1 and ICAM-1 were higher in normal peritoneum samples obtained from women with endometriosis compared to those from controls (P = 0.038 and P = 0.009). The mRNA levels of VCAM-1 were also higher in the eutopic endometrium samples obtained from women with endometriosis compared to controls (P = 0.018). With respect to serum protein levels, compared to controls, the women with endometriosis had lower serum levels of sICAM-1 (P = 0.042) and higher levels of sVCAM-1 (P < 0.001). Our analysis revealed that the serum levels of sVCAM-1 were not affected by lesion entity, menstrual cycle phase or disease severity. An receiver operating characteristics curve, calculated to determine whether preoperative serum sVCAM-1 concentration can be used to predict endometriosis, found an AUC of 0.868 with 80% specificity and 84% sensitivity at a cutoff value of 370 pg/ml. This predictive performance can be further improved by calculation of the sVCAM-1/sICAM-1 ratio, leading to an AUC of 0.929 with 86.7% specificity and 90.3% sensitivity at a cutoff ratio value of 1.55. Not applicable. The relatively small sample size in the expression analyses is a possible limitation of this study. Our findings could contribute to an improved understanding of the pathogenesis of endometriosis and the role of cell adhesion molecules. In addition, the results may lead to the development of new, non-invasive tools for diagnosing endometriosis. The ability to diagnose patients by measuring serum sVCAM-1 levels or the sVCAM-1/sICAM-1 ratio would have considerable clinical value. The Ingrid Flick Foundation (Grant no. FA751C0801), which played no role in the study design, data collection and analysis, decision to publish or preparation of the manuscript. The authors declare no competing interests.

Soluble Adhesion Molecules During Ex Vivo Lung Perfusion Are Associated With Posttransplant Primary Graft Dysfunction.

Ex vivo lung perfusion (EVLP) enables assessment of marginal donor lungs for transplantation. We aimed to discover biomarkers in EVLP perfusate that could predict development of primary graft dysfunction (PGD). From September 2008 to August 2013, 100 clinical EVLPs were performed. Eleven patients developed PGD grade 3 within 72 h after transplant. The non-PGD group consisted of 34 patients without PGD grade 3. Nonbilateral lung transplants or transplant after extracorporeal life support were excluded from analyses. Soluble intercellular adhesion molecule 1 (sICAM-1), soluble VCAM-1 (sVCAM-1), and soluble E selectin (sE-selectin) levels, as markers of endothelial activation, were measured in the perfusate of EVLP by enzyme-linked immunosorbent assay and were correlated with clinical outcome. Levels of sICAM-1 at 1 h and sVCAM-1 at 1 and 4 h were significantly higher in the PGD group compared with the non-PGD group. The sE selectin levels were not statistically different between the study groups. Higher levels of sVCAM-1 at 1 and 4 h were statistically significantly associated with PGD either alone or after adjustment for other PGD risk factors. These adhesion molecules may help identify donor lungs at higher risk of PGD during EVLP.

Endothelial function impairment in STEMI patients with out-of-hospital cardiac arrest under therapeutic hypothermia treatment

BackgroundTherapeutic hypothermia (HT) in out-of-hospital cardiac arrest STEMI patients aims to improve their neurological prognosis, but it has been associated with slow coronary flow and cardiac thrombotic events. We sought to serially assess endothelial function during the first 48h after admission in out-of-hospital cardiac arrest STEMI patients, under therapeutic hypothermia (HT). MethodsFrom January 2015 to August 2015, eighteen consecutive out-of-hospital cardiac arrest STEMI patients eligible for primary PCI received HT at admission and were included in the study (HT group). During the same time period, eight consecutive patients with large anterior STEMI who received primary PCI but not HT were included as control group. Serial endothelial function by measuring flow-mediated dilatation (FMD) in the brachial artery, biomarkers of endothelial function and oxidative stress were assessed during the first 48h after admission in both groups. ResultsHT group showed worse FMD as compared to the control group (p<0.001). Glutathione peroxidase-3 (GPx-3) values were higher in control as compared to HT group (p=0.019), without any interaction between time of observation and HT (p=0.864). A significant interaction between time and HT was found in the levels of sVCAM-1, which reached an earlier peak in control than in HT group (p=0.019). ET-1 values generally increase overtime (p=0.005), but without any main effect of HT (p=0.175). ConclusionsHT is associated with endothelial dysfunction in out-of-hospital cardiac arrest STEMI patients during the first 48h after admission. This vascular dysfunction may be related to increased oxidative stress due to deficiency of GPx-3 in HT patients.

GENETIC PREDICTORS OF INTENSIVE LIPID-LOWERING THERAPY EFFICACY AND ITS ANTI-INFLAMMATORY EFFECTS IN VERY HIGH CARDIOVASCULAR RISK PATIENTS

Aim . To study the anti-inflammatory effects of intensive lipid-lowering therapy and genetic predictors of its effectiveness in patients with very high cardiovascular risk. Material and methods . 58 patients with a history of cardiovascular disease and low-density lipoprotein cholesterol (LDL-C)>1.8 mmol/l or non-highdensity lipoprotein cholesterol (non-HDL-C)>2.6 mmol/l (mean age 61.9±8.6 (M±SD) years, 62.1% of men) were included into the study. Polymorphism Phe189Ser of the CYP3A4 (gene encoding cytochrome P4503A4, CYP3A4*17, rs4987161) and APOA1 (gene encoding apolipoprotein A): -75G/A in the promoter region (rs670) and +83C/T in the 5'-untranslated region (rs5069) was determined in all patients. Inflammatory status (high-sensitivity C-reactive protein (hsCRP), monocyte chemoattractant protein (MCP-1) and soluble vascular adhesion molecule (sVCAM-1)) was assessed initially and after 4 weeks of treatment with atorvastatin 80 mg/day. The results were considered statistically significant at p<0.05. Results . 16 (27.6%) patients achieved LDL-C<1.8 mmol/l, 21 (37.6%) patients – non-HDL-C<2.6 mmol/l. The carriership of the allele -75G of the APOA1 gene was associated with failure to achieve the target level of the non-HDL-C (Fisher's exact test p<0.05 (bilateral), p<0.01 (one-sided)). The levels of hsCRP, MCP-1 and sVCAM-1 significantly decreased by 46.8, 19.2 and 13.2%, respectively (p=0.0001). In the ROC analysis the level of MCP-1<471 pg/ml predicted the achievement of the target level of non-HDL-C with a sensitivity of 53.3% and a specificity of 90%. Conclusion . Intensive lipid-lowering therapy in patients with a very high cardiovascular risk is accompanied by a pronounced anti-inflammatory effect. The baseline level of MCP-1<471 pg/ml is associated with the achievement of the target values of non-HDL-C. The carriership of the allele (-75)G of the APOA1 gene is associated with the lack of achievement of the target level of non-HDL-C during intensive lipid-lowering therapy and can be considered as a predictor of resistance to statins.

Vascular Cell Adhesion Molecule 1, Intercellular Adhesion Molecule 1, and Cluster of Differentiation 146 Levels in Patients with Type 2 Diabetes with Complications.

BackgroundType 2 diabetes mellitus (T2DM) is a multisystemic, chronic disease accompanied by microvascular complications involving various complicated mechanisms. Intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and cluster of differentiation-146 (CD146) are mainly expressed by endothelial cells, and facilitate the adhesion and transmigration of immune cells, leading to inflammation. In the present study, we evaluated the levels of soluble adhesion molecules in patients with microvascular complications of T2DM.MethodsSerum and whole blood samples were collected from 58 T2DM patients with microvascular complications and 20 age-matched healthy subjects. Levels of soluble ICAM-1 (sICAM-1) and soluble VCAM-1 (sVCAM-1) were assessed using enzyme-linked immunosorbent assay, while flow cytometry was used to determine CD146 levels.ResultsSerum sICAM-1 levels were lower in T2DM patients with microvascular complications than in healthy controls (P<0.05). No significant differences were found in sVCAM-1 and CD146 levels between the study and the control group. Although patients were subdivided into groups according to the type of microvascular complications that they experienced, cell adhesion molecule levels were not correlated with the complication type.ConclusionIn the study group, most of the patients were on insulin therapy (76%), and 95% of them were receiving angiotensin-converting enzyme (ACE)-inhibitor agents. Insulin and ACE-inhibitors have been shown to decrease soluble adhesion molecule levels via various mechanisms, so we suggest that the decreased or unchanged levels of soluble forms of cellular adhesion molecules in our study group may have resulted from insulin and ACE-inhibitor therapy, as well as tissue-localized inflammation in patients with T2DM.

Достижение основных целей терапии при применении тройной фиксированной комбинации амлодипина, индапамида и периндоприла (препарат Ко-Дальнева®) у пациентов с артериальной гипертензией

Introduction. Combined antihypertensive therapy is indicated in 94% of patients with hypertension. Fixed combinations are preferable to use as they have a high profile of effectiveness, safety, more convenient to use and significantly increase the adherence. Objective. To evaluate the effectiveness of triple fixed combination of amlodipine/indapamide/perindopril for the control of blood pressure in patients who did not achieve the target blood pressure level in the previous combined therapy. Materials and methods. The study included 44 patients with hypertension of high and very high risk of cardiovascular complications (mean age - 58.59±10.57 years, 17 men, 27 women). Blood pressure levels, quality of life, anxiety and depression levels, adherence, biochemical indices, levels of inflammation markers (CRP, IL-6, IL-10) and endothelial dysfunction (sVCAM-1, VEGF) were assessed in all patients at baseline and after 6 months of amlodipine/indapamide/perindopril therapy. Results. Against the backdrop of the treatment, all patients reached the target level of blood pressure (BP), pulse BP decreased by 30.3 mm Hg. (p<0.001), all quality of life indicators improved: significant changes (p<0.05) in physical functioning (from 58.1±27.7 to 71.3±20.5%), role-physical functioning (from 54.7±41.2 to 85.5±31.5%), role-emotional (from 53.3±46.0 to 77.3±40.1%), vitality (from 53,4±16.9 to 63.2±14.5%). The anxiety levels (from 5.9±2.8 to 3.5±2.6) and depression (from 5.9±2.5 to 3.7±2.1) significantly decreased. Adherence increased to a high level (from 3.4±1.8 to 6.5±1.0 points, p<0.01). There were no reliable dynamics of biochemical parameters and markers of inflammation. The level of sVCAM-1 decreased (from 1063.5±442.4 to 898.67±433.5 ng/ml, p<0.001). There were no cases of orthostatic hypotension. Conclusion. Therapy with a triple fixed combination of perindopril/indapamide/amlodipine (Ko-Dalneva®) achieves the target blood pressure level with a significant reduction in pulse BP, increases adherence and quality of life, reduces anxiety and depression with good tolerability and metabolic neutrality, and improves the vascular endothelial state, significantly lowering sVCAM-1.

SVCAM-1, sICAM-1, TNF-α and IL-6 levels in bipolar disorder type I: Acute, longitudinal and therapeutic implications

Objectives: To explore the serum levels of soluble vascular cell adhesion molecule-1 (sVCAM-1), soluble intercellular cell adhesion molecule-1 (sICAM-1), tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in patients with bipolar disorder (BD), with regard to acute episode characteristics, course of the disorder and treatment.Methods: The study group consisted of 83 patients diagnosed with BD type I. The control group consisted of 73 healthy individuals, matched with the study group according to age, gender and body mass index. The serum levels of sVCAM-1, sICAM-1, TNF-α and IL-6 were measured by ELISA.Results: Compared with healthy controls, significantly elevated levels of IL-6 and sICAM-1 and significantly lower levels of TNF-α and sVCAM-1 were identified in acute and remission phases of BD. The acute serum levels of sVCAM-1 were associated with the type and severity of acute mood symptoms as well as with course of illness characteristics. TNF-α was associated with duration of untreated disorder and type of treatment.Conclusions: BD is related to both acute and long-term alterations of immune mediators, including adhesion molecules. The potential immunomodulatory role of pharmacotherapeutic treatment is also to be considered in BD.

Translation from SCD Mouse to Man: Chronic Administration of Tucaresol Reduces RBC Sickling and Improves Hematological Parameters, but Does Not Impact Inflammatory Markers in SCD Model Mice

Sickle cell disease is a hematological disorder with significant unmet need that affects approximately ten million people worldwide. Currently, treatment for SCD is limited to a single approved drug, Hydroxyurea, and palliative treatments such as hydration and pain management. The limited treatment options have led to considerable interest in developing new therapies for SCD. One strategy for a disease-modifying treatment for SCD is stabilization of the oxygenated form of hemoglobin (OxyHb) by a small molecule allosteric modulator (Zaugg et. Al, JBC, 1977). One such compound, Tucaresol was advanced into clinical trials on the strength of the pre-clinical data, however a SCD mouse model was unavailable when the trial was initiated. The renewed interest in stabilizing OxyHb with a small molecule presents an opportunity to use Tucaresol as a mechanism for understanding the translation between effects seen in an animal model and clinical endpoints in patients. In this study, we treated Townes SCD mice with Tucaresol for 15 days, to evaluate its effect on hematology, oxygen affinity, sickling and markers of inflammation in vivo. These results are compared to the clinical data reported in the original trials.Tucaresol is a substituted benzaldehyde derived from a class of molecules known to preferentially bind to hemoglobin and stabilize the oxygenated state by formation of a Schiff base linkage at a site on the a-chains (Rolan et. Al, Br. J. Clin. Pharm., 1995). Earlier molecules acting by this mechanism had been shown to reduce Hb S polymerization and RBC sickling in vitro, and it was theorized that long-term treatment with such molecules would impact hemolysis and occurrence of vaso-occlusive crisis (VOC) (Beddell et. Al, Br. J. Pharmac. 1984). Preclinical studies demonstrated Tucaresol modification of Hb S delays the polymerization process and inhibits RBC sickling. In clinical trials, Tucaresol positively impacted hematological markers including hemoglobin and reticulocyte counts, and reduced the percentage of irreversibly sickled cells. However, trials of Tucaresol were halted due to a significant immune response and specific inflammatory markers were not measured during the trial.To study these effects in Townes SCD model, mice were dosed with Tucaresol at 30 mg/kg twice a day, for fifteen days. The dose was selected to be consistent with clinical dosing regimens. Blood was collected prior to study start from individual animals to obtain baseline levels for hematological parameters and soluble cell adhesion molecules in plasma. Blood was sampled at day 3 and day 10 of dosing to understand drug exposure and accumulation. At this dose, Tucaresol demonstrated a steady state achieved by day 10 and the blood concentration ranged from 0.5-1.6 mM on day 15. Following two week dosing with Tucaresol, we observed a 50% increase in hemoglobin, and 24% increase in mean corpuscular hemoglobin concentration (MCHC), relative to vehicle treated animals. Reticulocytes, a clinical indicator of hemolytic anemia, are highly elevated in SCD patients and mice, also showed a 47% decrease in Tucaresol treated relative to vehicle treated animals. Oxygen affinity measurements showed a decrease in P50 and P20 values for treated animals, indicating increased oxygen affinity, consistent with the proposed mode of action of Tucaresol- stabilization of the oxygenated state. The percentage of sickled erythrocytes was also decreased by 38% in Tucaresol treated SCD mice when compared to vehicle controls. The decrease in sickling is consistent with the clinical observation of a 52% (mean) decrease in irreversibly sickled cells. In contrast to the improvement seen with hematological parameters, inflammatory markers remained either unchanged or slightly elevated after 15 days of treatment with Tucaresol. Plasma levels of sICAM-1 and sVCAM-1 were unchanged while sE-Selectin was elevated. In summary, in this short course of treatment of SCD mice, Tucaresol had an impact on the hematological markers that was consistent with effects observed in the clinic, but did not modulate the inflammation response.All experiments were within guidelines and were reviewed and approved by Pfizer institutional animal care and use committee. DisclosuresKnee:Pfizer Inc: Employment. Jasuja:Pfizer: Employment. Barakat:Pfizer: Employment. Kelly:Pfizer: Employment. Singh:Pfizer Inc: Employment. Janz:Pfizer: Employment.

Systemic Heme and Iron Overload Results in Depletion of Serum Hemopexin, Haptoglobin and Transferrin and Correlates with Markers of Endothelial Activation and Lipid Oxidation in Beta Thalassemia Major and Intermedia

Beta thalassemia is an inherited hemoglobinopathy due to reduced synthesis of Beta globin chains and, consequently, of hemoglobin A (a2b2). The clinical manifestations are mainly the result of chronic anemia and iron overload. The latter is due to increased iron absorption, induced by accelerated but ineffective erythropoiesis, and recurrent red blood cell transfusions. Alfa-chains and iron excess promote oxidative damage of red blood cell membrane, resulting in macrophage sequestration and extravascular hemolysis, and to a lower extent, in intravascular hemolysis, with consequent release of hemoglobin (Hb), heme and iron.Increasing evidence suggests that free heme exerts vasculotoxic, pro-inflammatory and procoagulant effects due to its ability to trigger endothelial and immune cells activation. In addition, a role for heme and iron has been postulated in the pathogenesis of other vascular diseases, including atherosclerosis. In mouse models of Beta thalassemia and sickle cell disease, circulating heme levels are elevated and correlate with the exhaustion of systemic scavengers for hemoglobin and heme, haptoglobin and hemopexin, respectively, as well as with severe endothelial dysfunction and inflammation. Hemopexin-based therapies significantly improve endothelial damage, vascular oxidative stress and inflammation in these mice (Vinchi et al., Circulation 2013, Blood 2016; Vercellotti GM. et al., Mol Med 2016).Whereas more data are reported on sickle patients in this regard, few data are available in patients with Beta thalassemia. In the present study, we examined serum samples from a cohort of 60 patients with Beta thalassemia major (age 11.5 ± 6.8, 44% males-56% females, Hb 7.69 ± 1.22 mg/dl, transfused every 3-4 weeks) and 7 patients with Beta thalassemia intermedia (age 14 ± 12 , 70% males-30% females, Hb 8.4 ± 0.74 mg/dl, transfused every 4-5 weeks). 10% of the patients received inconsistent iron chelation therapy. Serum from 10 healthy subjects (age 22.7±15.3, 50% males-50% females, Hb 13.12±1.15 mg/dl) served as control.Both groups of patients show high systemic heme and iron levels, which associate with a severe drop in serum haptoglobin, hemopexin and transferrin. Consistently, transferrin saturation (12.4±2 vs 79.6±24 %) and serum ferritin (55.14 ±0.23 vs 4919.2 ±2657.4 ng/ml) are elevated. Interestingly, these patients present with high systemic levels of the soluble adhesion molecules sVCAM-1 and sICAM-1, markers of enhanced endothelial activation. In addition, they show increased levels of serum malondialdehyde, a well-known marker of lipid peroxidation and oxidative stress, and high levels of circulating oxidized low density lipoproteins (oxLDL). All parameters significantly correlate with increased systemic heme and iron indices as well as decreased haptoglobin, hemopexin and transferrin levels.In conclusion, Beta thalassemia patients show a strong correlation between systemic heme and iron overload, depletion of the respective scavengers, and markers of oxidative stress and endothelial dysfunction, thus confirming studies in animal models. These results emphasize the involvement of serum hemoglobin, heme and iron in the pathophysiology of Beta thalassemia, including vascular dysfunction, and the key protective role of their carriers. These findings are relevant for disorders hallmarked by vasculopathy, such as sickle cell disease and Beta thalassemia, as well as cardiovascular diseases, such as atherosclerosis. Our data support the potential therapeutic benefit of the administration of hemoglobin/heme scavengers along with efficient iron chelation therapy to counteract heme- and iron-driven toxicity.(The last three authors equally contributed to the work)****P<0.0001 [Display omitted] DisclosuresVercellotti:CSL-Behring: Research Funding; Imara: Research Funding. Belcher:Cydan/Imara: Research Funding; CSL-Behring: Research Funding.

Impact of CYP2C19 Variants on Clinical Efficacy of Clopidogrel and 1-Year Clinical Outcomes in Coronary Heart Patients Undergoing Percutaneous Coronary Intervention

The impact of pharmacogenetic variants of cytochrome P450 2C19 (CYP2C19) on clopidogrel-mediated effects on platelet inhibition, inflammatory response and endothelial function, as well as risk of major adverse cardiovascular events (MACE), in coronary heart patients undergoing percutaneous coronary intervention (PCI) was investigated. To this end, we assessed the residual platelet aggregation rate (RPA), maximal aggregation rate (MAR) and plasma levels of sCD40L, sP-selectin, MMP-9, sVCAM-1 and sE-selectin after 24 h of PCI in 559 patients treated with clopidogrel and followed up for 1 year for evidence of MACE. CYP2C19 *2 and *3 variants were identified using a clopidogrel-sensitive gene detection kit. Our results showed higher RPA and MAR as well as increased sE-selectin, sCD40L, sP-selectin, MMP-9, and sVCAM-1 levels in CYP2C19 intermediate metabolizer (IM, CYP2C19*1/*2, or *1/*3), poor metabolizer (PM, CYP2C19*2/*2, *2/*3, or *3/*3) and combined IM+PM groups, relative to those in extensive metabolizers (EM, CYP2C19*1/*1). In total, 519 patients completed 1 year of follow-up, among which 69 (13.3%) experienced MACE. The risk of MACE in CYP2C19 IM+PM patients was 2.664 times higher than that in CYP2C19 EM patients (OR = 2.664 (1.397–5.193), P = 0.004). The data suggest that CYP2C19*2 and *3 variants modulate the drug efficacy of clopidogrel in coronary heart patients undergoing PCI and further enhance the risk of MACE. Accordingly, CYP2C19 pharmacogenetic profiling may be beneficial for coronary heart patients undergoing PCI to predict the efficacy of treatment with clopidogrel. We propose that IM and PM patients should benefit from treatment with higher clopidogrel doses to improve efficacy and reduce the incidence of MACE.

Cytokine profiles in pediatric multiple sclerosis

The immunopathogenesis of pediatric multiple sclerosis (MS) is not well understood. We studied the cytokine profile in pre-treatment serum specimens of 19 pediatric MS patients, 25 adult MS patients, and 22 age- and gender-matched pediatric healthy controls. In addition to IL-2, IL-12p40, IL-12p70, IL-18, IL-23, IL-6, TNF-α, TGF-β-1, IFN-γ, IL-17A, IL-21, IL-10, IL-4, IL-5, IL-13, and GM-CSF, we measured osteopontin and soluble VCAM-I. In children with MS, significantly lower levels of IL-6 were present compared to age- and gender-matched healthy control children (p < 0.05). Moreover, significantly higher levels of osteopontin (p < 0.02) and sVCAM-1 (p < 0.02) and lower levels of IL-6 (p < 0.01) were present, with trends toward lower levels of IL-12p70 (p = 0.074) and IL-17a (p = 0.05) compared to adults with MS. These findings indicate important differences in cytokine signatures in children with MS and suggest an unexpected possible lower inflammatory cytokine profile in children with MS.

Circulating levels of MCP-1, VEGF-A, sICAM-1, sVCAM-1, sE-selectin and sVE-cadherin: Relationship with components of metabolic syndrome in young population

IntroductionInflammation and endothelial dysfunction are considered the primary manifestations of the cardiovascular disease. Studies have established a relationship among components of metabolic syndrome (MetS) with inflammatory markers and the loss of permeability, vasoconstriction and vasodilatation endothelial. ObjectiveTo determine the relationship among the concentrations of soluble endothelial dysfunction molecules and inflammation cytokines and components of the metabolic syndrome in young population. Material and methodsA study was performed in 240 young adult students ages 18–28 years. To define the presence of clinical and metabolic alterations and MetS the modified ATP-III criteria was considered. In all subjects were determined sociodemographic characteristics, anthropometric measures and the metabolic profile. Circulating levels of MCP-1, VEGF-A, sICAM-1, sVCAM-1, sE-selectin and sVE-cadherin were determined by ELISA immunoassay (Bioscience). Statistical analysis was performed using STATA statistical software v. 9.2. ResultsFrom all the participants, 44.6% had obesity, 59.9% had abdominal obesity, 49.6% low HDL-c and 16.7% high levels triglycerids. The 16.25% of the population showed 3 or more components of the MetS. Elevated MCP-1, sICAM-1 and sE-selectin levels were linked to the presence of obesity. In a model adjusted by age–gender, high soluble levels of MCP-1 and VEGF-A were linked with abdominal obesity (OR=1.83; 1.02–3.28 and OR=2.03; 1.15–3.56, respectively), as well as to the presence of the 2 components of MetS. sVCAM-1 levels were associated with impaired glucose (OR=4.74; 1.32–17.0); sE-selectin with low HDL-c (OR=1.99; 1.05–3.75), although sICAM-1 and sVE-cadherin were associated with impaired systolic blood pressure (OR=4.04; 1.24–13.1 and OR=6.28; 1.90–20.7, respectively). ConclusionLevels of circulating MCP-1 and VEGF-A were associated with adiposity, levels of sVCAM-1 with the presence of impaired glucose, sE-selectin with low HDL-c, while the levels of sICAM-1 and sVE-cadherin were associated with impaired systolic blood pressure in young adults independently of other traditional risk factors.

Soluble vascular cell adhesion molecular-1 is a potential biological indicator of hemophilic arthropathy.

Hemophilic arthropathy is the most common chronic complication in patients with hemophilia. The pathogenesis of hemophilic arthropathy involves the inflammatory processes associated with rheumatoid arthritis (RA). Determining the severity and/or progression of joint damage is crucial when evaluating the effect of treatment modalities. Identifying reliable biomarkers in the peripheral blood of patients with hemophilic arthropathy may be beneficial in clinical practice. Circulating soluble vascular cell adhesion molecule-1 (sVCAM-1), E-selectin, and P-selectin levels are elevated in patients with RA. Our study investigated whether these soluble adhesion molecules can be used as biological indicators in the course of joint damage in patients with hemophilia A.Patients with hemophilia A (mild, moderate, and severe) were enrolled. The plasma levels of sVCAM-1, E-selectin, and P-selectin in patients with hemophilia A and control were measured using specific enzyme-linked immunosorbent assay kits. Joint damages were evaluated using Pettersson scores.No statistically significant differences were observed in E-selectin and P-selectin levels between patients and controls. The sVCAM-1 level was significantly higher in patients with hemophilia A than in controls. The differences remained significant in patients with severe hemophilia A but not in patients with mild or moderate hemophilia A. The degree of hemophilic arthropathy was evaluated using Pettersson scores, and a score higher than 5 indicated marked arthropathy. Patients with more than 1 joint with marked arthropathy showed significantly higher sVCAM-1 levels.sVCAM-1 levels in patients with hemophilia A are associated with the severity of hemophilic arthropathy.

Leptin promotes endothelial dysfunction in chronic kidney disease through AKT/GSK3β and β-catenin signals

Endothelial dysfunction (ED) is a well-recognized instigator of cardiovascular diseases and develops in chronic kidney disease (CKD) with high rate. Recent studies have implicated that leptin is associated with endothelial dysfunction. We investigated the relationship between leptin and markers of ED in CKD patients and how leptin contributed to endothelial damage. 140 CKD patients and 140 healthy subjects were studied. Serum leptin levels were significantly higher in CKD than in controls and displayed significantly positive association with the increase levels of sICAM-1 and sVCAM-1 but negative correlation with flow-mediated dilatation (FMD) reduction in patients. Our in vitro study demonstrated that leptin induced overexpression of ICAM-1 and VCAM-1, led to f-actin reorganization and vinculin assembly, increased endothelial monolayer permeability for FITC-dextran, and accelerated endothelial cell migration; these changes were markedly reversed when the cells were transfected with AKT or β-catenin shRNA vectors. Notably, high leptin resulted in hyper-phosphorylation of AKT and GSK3β, along with nuclear accumulation of β-catenin. In conclusion, serum leptin was elevated in CKD patients and it might contribute to endothelial dysfunction by disarrangement of f-actin cytoskeleton via a mechanism involving the AKT/GSK3β and β-catenin pathway.

Serum sPECAM-1 and sVCAM-1 levels are associated with conversion to multiple sclerosis in patients with optic neuritis

Platelet-Endothelial-Cell-Adhesion-Molecule-1 (PECAM-1) and Human-Vascular-CAM-1 (VCAM-1) are adhesion molecules involved in leukocyte-endothelial interaction. In our study serum levels of sPECAM-1 and sVCAM-1 were measured (ELISA) in twenty-nine patients during their first monosymptomatic optic neuritis (ON) episode. Anti-aquaporin-4-antibodies (AQP4-IgG) were detected with the cell-based assay. Patients were followed for seven years, during which 16/24 AQP4-IgG (−) patients developed MS and 2/5 AQP4-IgG (+) patients developed NMO. Patients who developed MS had significantly lower sPECAM-1 and sVCAM-1 than those who did not. Serum sPECAM-1 and sVCAM-1 may turn out to be useful biomarkers correlated with the risk of progression to MS after first ON incident.

P.3.f.021 - Acute paranoid schizophrenia patients present higher serum S100B protein levels at admission than at discharge

Schizophrenia (SZ) is a neuroprogressive disorder presenting with biochemical, functional, and structural changes, which differ from early to late stages of the illness. We explored the differences in serum levels of soluble intercellular cell adhesion molecule-1 (sICAM-1) and soluble vascular cell adhesion molecule-1 (sVCAM-1) between early and late stages of SZ, in regard to clinical characteristics and treatment application.Serum levels of sICAM-1 and sVCAM-1 were measured in 80 patients with SZ (40 early stage; 40 late stage), and compared with 80 healthy controls, matched by age, gender, body mass index, and smoking habits with each SZ group. Serum levels of sICAM-1 and sVCAM-1 were measured using ELISA. The severity of psychopathology was assessed using the Clinical Global Impression Scale and five-factor Positive and Negative Symptoms in Schizophrenia Scale.After adjustment for confounders, we noticed normal levels of sICAM-1 in the early stage, and elevated levels of sICAM-1 in the late stage of SZ. sVCAM-1 levels were decreased in both stages of SZ. Higher sICAM-1 levels have been related to more pronounced cognitive deficit and excitement symptoms in the early stage of SZ and to favorable characteristics of treatment application in both stages.SZ is associated with changes in the levels of adhesion molecules that vary from early to late stages of the illness. This implies that the concept of biochemical staging is applicable in SZ, at least for markers of cellular adhesion.

Soluble vascular cell adhesion molecule-1 and magnesium sulfate with nifedipine treatment in Indonesian women with severe pre-eclampsia.

Endothelial cell activation in pre-eclampsia is associated with elevated soluble vascular cell adhesion molecule-1 (sVCAM-1) levels. The objective of the study was to determine whether sVCAM-1 levels in Indonesian women with pre-eclampsia were similar to other ethnic studies and to determine the effects of magnesium sulfate with nifedipine on blood pressure. A total of 61 pregnant women were admitted, who had normal pregnancy (n = 25) and severe pre-eclampsia (n = 36). Blood sampling was performed at admission to the study, 1h after placental separation, and 24h postpartum. sVCAM-1 and blood pressure levels were determined. The mean ages in normal pregnancy (n = 25) and in severe pre-eclampsia (n = 36) are 30.0 ± 3.4 years and 27.1 ± 6.1 years, respectively. Significantly elevated sVCAM-1 was seen in pre-eclampsia. No significant variation in sVCAM-1 levels during the study periods was seen in both groups of cohorts. Magnesium sulfate infusion and nifedipine significantly lowered the blood pressure level. Elevated sVCAM-1 levels were also seen in Indonesian women with severe pre-eclampsia. The placenta may not be the only source of elevated sVCAM-1 and that endothelial dysfunction persists beyond the postpartum period. Magnesium sulfate together with nifedipine significantly lowered blood pressure. The determination of elevated sVCAM-1 in pregnancy as a risk marker for endothelial dysfunction is therefore suggested.