Effect of Orally-administration Niclosamide on serum level of VCAM-1 and E-selectin in Collagen induced arthritis: Experimental study

Soluble adhesion molecules (mainly VCAM-1 and E-selectin) have a vital role in the pathogenesis of the rheumatoid arthritis (RA) and consider as angiogenic mediators for this disease. The main goal for this research is to evaluate the efficacy of orally administer niclosamide (NCS) in prevention the angiogenic mediators (VCAM-1 and E-selectin) using collagen induced arthritis model in rats (CIA). Fifty male Spraque-Dawley rats underwent collagen induced arthritis (CIA) model. When arthritis was fully developed, the rats were either treated orally with low-dose (50mg/kg) NCS or high-dose (100mg/kg) NCS or treated intrapertonially (IP) with 30mg/kg NCS or leave without treatment for 4 weeks. Body weight measurement and arthritis index were monitor before and after treatment in all groups. At the end of the treatment period serum level of vascular cell adhesion molecule-1 (VCAM-1), E-selectin and TNFα were measured together with collection of articular synovial tissue to evaluate the pathological changes. The experiment showed that NCS significantly reduce the arthritis index, foot pad thickness and ankle swelling (p value < 0.05) when given orally in a high dose and IP to the experimental animal. Comparing to the CIA model group, significant reduction in the serum level of VCAM1 and E-selectin has been observed in those rats treated with high dose of oral NCS or IP injection of NCS (p value < 0.05). Niclosamide can effectively decrease, in dose dependent manner, the clinical scores, joint swelling, VCAM1, E-selectin and pathological changes in arthritic rats induced by collagen type II.

P042 Diagnostic and prognostic value of soluble adhesion molecules in patients with ulcerative colitis

Background Experimental studies have shown that cell adhesion molecules contribute to the constant induction of pro-inflammatory cytokines (interleukin (6, 8, 10), necrosis factor of tumor-α), providing the chronicity of immune-mediated inflammation at ulcerative colitis (UC). Determination of the importance of molecules of integrin adhesion (sVCAM-1) and mucosal adressin (sMAdCAM-1) for evaluating the effectiveness of treatment of patients with UC. Methods 119 patients with UC were examined: 17 (14.3%) with proctitis, 44 (37%) with left-sided localization, 58 (48.7%) with the total form. The comparison group consisted of 20 healthy volunteers. Determination of serum sVCAM and sMAdCAM was carried out by quantitative enzyme-linked immunosorbent assay (ELISA) based on a system for multiplex analysis using flow fluorimetry — Luminex MAGPIX (USA), ICAM-1 kit (Cusabio, USA). The indicators were assessed before the start of the basic therapy course, in accordance with national recommendations, and 12 weeks after its completion. Results In patients with left-sided and total UC, a 3.2 and 4.7-fold increase in sVCAM expression and a 2.8, 3.6-fold increase in sMAdCAM expression was recorded, averaging: 386.6 + 21.2 ng/ml, 623.4 + 11.1 ng/ml; 116.5 + 13.6 ng/ml, 193.8 + 15.2 ng/ml, respectively (p &lt;0.001). In the group with rectal lesions, there was a moderate increase in the expression of adhesion molecules: 157.1 + 11.3 ng/ml, 84.5 + 18.8 ng/ml, respectively (p&lt;0.07). Against the background of a 12-week treatment course, clinical and endoscopic remission was recorded in 84 (70.6%) patients. In the groups of patients with left-sided and total UC in the drug remission phase, there was a decrease in the expression of sVCAM (194.6 + 9.2 ng/ml, 236.7 + 14.1 ng/ml) and sMAdCAM (72.4 + 8.1 ng/ml, 98.2 + 9.7 ng/ml), respectively (p &lt;0.02). In cases with UC activity, increased levels of adhesion molecules persisted. Conclusion The soluble adhesion molecules of sVCAM and sMAdCAM are modern markers of inflammation that can be used for assessing the effectiveness of course therapy of UC.

The Constitutive Extracellular Protein Release by Acute Myeloid Leukemia Cells—A Proteomic Study of Patient Heterogeneity and Its Modulation by Mesenchymal Stromal Cells

Extracellular protein release is important both for the formation of extracellular matrix and for communication between cells. We investigated the extracellular protein release by in vitro cultured normal mesenchymal stem cells (MSCs) and by primary human acute myeloid leukemia (AML) cells derived from 40 consecutive patients. We observed quantifiable levels of 3082 proteins in our study; for the MSCs, we detected 1446 proteins, whereas the number of released proteins for the AML cells showed wide variation between patients (average number 1699, range 557–2380). The proteins were derived from various cellular compartments (e.g., cell membrane, nucleus, and cytoplasms), several organelles (e.g., cytoskeleton, endoplasmatic reticulum, Golgi apparatus, and mitochondria) and had various functions (e.g., extracellular matrix and exosomal proteins, cytokines, soluble adhesion molecules, protein synthesis, post-transcriptional modulation, RNA binding, and ribonuclear proteins). Thus, AML patients were very heterogeneous both regarding the number of proteins and the nature of their extracellularly released proteins. The protein release profiles of MSCs and primary AML cells show a considerable overlap, but a minority of the proteins are released only or mainly by the MSC, including several extracellular matrix molecules. Taken together, our observations suggest that the protein profile of the extracellular bone marrow microenvironment differs between AML patients, these differences are mainly caused by the protein release by the leukemic cells but this leukemia-associated heterogeneity of the overall extracellular protein profile is modulated by the constitutive protein release by normal MSCs.

Plasma Biomarkers of Risk of Tuberculosis Recurrence in HIV Co-Infected Patients From South Africa

There is an urgent need to identify immunological markers of tuberculosis (TB) risk in HIV co-infected individuals. Previously we have shown that TB recurrence in HIV co-infected individuals on ART was associated with markers of systemic inflammation (IL-6, IL1β and IL-1Rα). Here we examined the effect of additional acute inflammation and microbial translocation marker expression on risk of TB recurrence. Stored plasma samples were drawn from the TB Recurrence upon Treatment with HAART (TRuTH) study, in which individuals with previously treated pulmonary TB were screened for recurrence quarterly for up to 4 years. Recurrent TB cases (n = 37) were matched to controls (n = 102) by original trial study arm assignment and ART start date. Additional subsets of HIV infected (n = 41) and HIV uninfected (n = 37) individuals from Improving Recurrence Success (IMPRESS) study were sampled at active TB and post successful treatment completion. Plasma concentrations of soluble adhesion molecules (sMAdCAM, sICAM and sVCAM), lipopolysaccharide binding protein (LBP) and transforming growth factor-beta (TGF-β1, TGF-β2, TGF-β3) were measured by multiplex immunoassays and ELISA. Cytokine data was square root transformed in order to reduce variability. Multivariable analysis adjusted for a number of potential confounders measured at sample time-point: age, BMI, CD4 count, viral load (VL) and measured at baseline: presence or absence of lung cavities, previous history of TB, and WHO disease stage (4 vs 3). The following analytes were associated with increased risk of TB recurrence in the multivariable model: sICAM (aOR 1.06, 95% CI: 1.02-1.12, p = 0.009), LBP (aOR 8.78, 95% CI: 1.23-62.66, p = 0.030) and TGF-β3 (aOR 1.44, 95% CI 1.01-2.05, p = 0.044). Additionally, we observed a positive correlation between LBP and sICAM (r= 0.347, p&lt;0.0001), and LBP and IL-6, identified to be one of the strongest predictors of TB risk in our previous study (r=0.623, p=0.03). These data show that increased risk of TB recurrence in HIV infected individuals on ART is likely associated with HIV mediated translocation of microbial products and the resulting chronic immune activation.

Atheroprotective Properties of Costus spicatus (Jacq.) Sw. in Female Rats

Background: Costus spicatus (Jacq.) Sw. is a medicinal species frequently prescribed for the treatment of cardiovascular diseases. This study aims to evaluate the effects of this species against the development of atherosclerosis. Methods: First, an anatomical study of the C. spicatus leaves was performed. Then, the extract (ESCS) was obtained and submitted to phytochemical analysis. Female rats were treated with a single dose of ESCS (2000 mg/kg) to assess acute toxicity. Other groups of female rats received an atherogenic diet for 60 days. After 30 days, the animals were treated orally with ESCS (30 and 300 mg/kg), rosuvastatin (5 mg/kg), or vehicle once daily for 30 days. Serum lipids oxidized low-density lipoprotein, soluble adhesion molecules, interleukins 1β and 6, and markers of renal and liver function were measured. Renal function, blood pressure, electrocardiography, and vascular reactivity were also evaluated. Arteries, heart, liver, and kidney were also collected to evaluate the tissue redox state and histopathological analysis. Results: Prolonged treatment with ESCS induces significant hypolipidemic and antioxidant effects, that prevent endothelial dysfunction and modulated the local inflammatory process, reducing the evolution of the atherosclerotic disease. Conclusions: This study provides a scientific basis for the popular use of C. spicatus for the treatment of atherosclerosis.

Elevated Pro-Inflammatory Cell-Free MicroRNA Levels in Cerebrospinal Fluid of Premature Infants after Intraventricular Hemorrhage

Intraventricular hemorrhage (IVH) represents a high risk of neonatal mortality and later neurodevelopmental impairment in prematurity. IVH is accompanied with inflammation, hemolysis, and extracellular hemoglobin (Hb) oxidation. However, microRNA (miRNA) expression in cerebrospinal fluid (CSF) of preterm infants with IVH has been unknown. Therefore, in the present study, candidate pro-inflammatory cell-free miRNAs were analyzed in CSF samples from 47 preterm infants with grade III or IV IVH vs. clinical controls (n = 14). miRNAs were quantified by RT-qPCR, normalized to “spike-in” cel-miR-39. Oxidized Hb and total heme levels were determined by spectrophotometry as well as IL-8, VCAM-1, ICAM-1, and E-selectin concentrations by ELISA. To reveal the origin of the investigated miRNAs, controlled hemolysis experiments were performed in vitro; in addition, human choroid plexus epithelial cell (HCPEpiC) cultures were treated with metHb, ferrylHb, heme, or TNF-α to replicate IVH-triggered cellular conditions. Levels of miR-223, miR-155, miR-181b, and miR-126 as well as Hb metabolites along with IL-8 were elevated in CSF after the onset of IVH vs. controls. Significant correlations were observed among the miRNAs, oxidized Hb forms, and the soluble adhesion molecules. During the post-IVH follow-up, attenuated expression of miRNAs and protein biomarkers in CSF was observed upon elimination of Hb metabolites. These miRNAs remained unaffected by a series of artificially induced hemolysis, which excluded red blood cells as their origin, while stimulation of HCPEpiCs with oxidized Hb fractions and heme resulted in increased extracellular miRNA levels in the cell culture supernatant. Overall, the hemorrhage-induced CSF miRNAs reflected inflammatory conditions as potential biomarkers in preterm IVH.