The Comparation of Activation Nuclear Factor Kappa Beta (NFKB) at Rattus Novergicus Strain Wistar Induced by Various Duration High Fat Diet (HFD)

NFκB is a transcription factor regulating many function of the vessel wall. In the normal condition , NFκB is revealed diffuse cytoplasmic expressionsuggesting that the system is inactive. The presence of activation NFκB provide a potential pathway for the rapid transcriptional of a variety of genes encoding cytokines, growth factors, adhesion molecules and procoagulatory factors. It is likely to play an important role in chronic inflamatory disease involved atherosclerosis. There are many stimuli with the potential to active NFκB, including hyperlipidemia. We used 24 mice which was divided in 6 groups. The HFD given by et libitum procedure during 2, 4, and 6 months. The parameters in this study were the amount of NFKB activation ,H2O2 as ROS and VCAM-1 as a product of NFKB activation. H2O2 colorimetryc assay performed directly using Anti Rat H2O2 ELISA Kit. The NFKB and VCAM-1 detection obtained from aorta mice, measured by ELISA kit and imunohistochemistry. There was a significant difference activation of H2O2, NFKB and VCAM-1 level at induce HFD after 2, 4 and 6 months. It suggest that HFD induce ROS formation and increase the activation of NFKB as one of atherosclerosis marker that caused by hyperlipidemia as classical atheroschlerosis risk factor.

Development of an Immunoassay Platform for Diagnosis of Acute Kidney Injury

Acute kidney injury (AKI) is a new worldwide public health problem. A diagnosis of this disease using creatinine is still a problem in clinical practice. Therefore, a measurement of biomarkers responsible for AKI has received much attention in the past couple years. Cytokine interleukin-18 (IL-18) was reported as one of the early biomarkers for AKI. The most commonly used method to detect this biomarker is an immunoassay. This study used a planar platform to perform an immunoassay using fluorescence for detection. In this study, anti-IL-18 antibody was immobilized onto a microscope slide using a covalent binding method. Make-up samples were diluted at the concentration between 10 to 1000 pg/ml to create a calibration curve. The precision of the system was determined using a coefficient of variability (CV), which was found to be less than 10%. The performance of this immunoassay system was compared with the measurement from ELISA.

In vivo Antidiabetic and Antioxidant Potential of Pseudovaria macrophylla Extract

This study has investigated the antidiabetic and antioxidant potential of Pseudovaria macrophylla bark extract on streptozotocin–nicotinamide induced type 2 diabetic rats. LCMSQTOF and NMR experiments were done to determine the chemical composition in the methanolic bark extract. For in vivo experiments, the STZ (60 mg/kg/b.w, 15 min after 120 mg/kg/1 nicotinamide, i.p.) induced diabetic rats were treated with methanolic extract of Pseuduvaria macrophylla (200 and 400 mg/kg·bw) and glibenclamide (2.5 mg/kg) as positive control respectively. Biochemical parameters were assayed in the blood samples of all groups of rats. The pro-inflammatory cytokines, antioxidant status and plasma transforming growth factor βeta-1 (TGF-β1) were evaluated. The histological study of the pancreas was examined and its expression level of insulin was observed by immunohistochemistry. In addition, the expression of glucose transporters (GLUT 1, 2 and 4) were assessed in pancreas tissue by western blot analysis. The outcomes of the study displayed that the bark methanol extract of Pseuduvaria macrophylla has potentially normalized the elevated blood glucose levels and improved serum insulin and C-peptide levels with significant increase in the antioxidant enzyme, reduced glutathione (GSH) and decrease in the level of lipid peroxidation (LPO). Additionally, the extract has markedly decreased the levels of serum pro-inflammatory cytokines and transforming growth factor beta-1 (TGF-β1). Histopathology analysis demonstrated that Pseuduvaria macrophylla has the potential to protect the pancreas of diabetic rats against peroxidation damage by downregulating oxidative stress and elevated hyperglycaemia. Furthermore, the expression of insulin protein, GLUT-1, GLUT-2 and GLUT-4 in pancreatic cells was enhanced. The findings of this study support the anti-diabetic claims of Pseudovaria macrophylla bark.

Changes of in vitro Cytokine Production induced by δ-Lactams

The aim of this work was to study the in vitro effects of δ-lactam 1 and its 4-chlorophenyl derivative 2, on the proliferative responses of human lymphocytes and Th1 and Th2 cytokine secretion. The possible protective role of vitamin E on intracellular stress oxidative induced by these compounds was also investigated. Peripheral blood lymphocytes were isolated using differential centrifugation on a density gradient of Histopaque. They were cultured with mitogen concanavalin A, vitamin E (10 μM) and with different concentrations of the compounds 1 and 2 (0.1 to 10 μM). Proliferation (MTT assay), IL-2, INFγ and IL-4 (Elisa kits), intracellular superoxide anion were determined. 1 and 2 were immunostimulant and increased cytokine secretion with a shift away from Th1 response to Th2. These properties were however accompanied by an increase in intracellular oxidative stress. The presence of vitamin E exhibited protective effects by reducing δ- lactam-induced superoxide anion generation in lymphocytes.

Highly Sensitive Label Free Biosensor for Tumor Necrosis Factor

We present a label-free biosensor based on electrochemical impedance spectroscopy for the detection of proinflammatory cytokine Tumor Necrosis Factor (TNF-α). Secretion of TNF-α has been correlated to the onset of various diseases including rheumatoid arthritis, Crohn-s disease etc. Gold electrodes were patterned on a silicon substrate and self assembled monolayer of dithiobis-succinimidyl propionate was used to develop the biosensor which achieved a detection limit of ~57fM. A linear relationship was also observed between increasing TNF-α concentrations and chargetransfer resistance within a dynamic range of 1pg/ml – 1ng/ml.