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Background: Oroxylum indicum (L.) Kurz is an edible plant that is widely consumed in several Asian countries. In our previous study, it was demonstrated that Oroxylum indicum (L.) Kurz seed (OIS) extract attenuated oxidative stress and proinflammatory cytokines in activated microglia. However, the molecular mechanism remains unknown. Objectives: To confirm the antiinflammatory and antioxidative effects of OIS extract on and molecular mechanisms of OIS in lipopolysaccharide (LPS)-induced microglial activation. Materials and Methods: Nitrite and Chloro methyl derivative of 2', 7'-dichlorodihydrofluoresceindiacetate (CM-H2DCFDA) assays were performed to examine the effect of OIS extract on LPS-induced nitric oxide (NO) and reactive oxygen species (ROS) production, respectively. Immunofluorescence and ELISA were performed to detect the levels of nuclear factor-κB (NF-κB) in the nucleus. Western blot analysis was performed to detect the phosphorylated (p-) form of Akt and extracellular-signal-regulated kinase 1/2 (ERK1/2). Results: OIS extract exhibited a significant and dose-dependent suppressive effect on LPS-induced NO and ROS production. Mechanistically, OIS extract attenuated the p-Akt and p-ERK1/2 signaling pathways, thereby inhibiting the LPS-induced NF-κB nuclear translocation. Conclusion: OIS extract exerts anti-oxidative effects by suppressing ROS production, and anti-inflammatory effects by inhibiting NO in activated BV2 microglial cells. Mechanistically, OIS extract could inhibit the Akt/ERK1/2-mediated NF-κB pathway. OIS extract may act as a promising functional food for the alleviation of neuroinflammation in neurodegenerative diseases. [ABSTRACT FROM AUTHOR]

Purpose: To investigate the anti-inflammatory effects and antioxidant activity of Oroxylum indicum (L.) Kurz fruit extract in lipopolysaccharide (LPS)-stimulated BV2 microglia. Methods: BV2 cells were treated with LPS for 24 h in the presence or absence of O. indicum fruit extract. Then, nitric oxide (NO), reactive oxygen species (ROS) and interleukin 6 (IL-16) levels were measured using Griess reagent assay, CM-H2DCFDA and enzyme-linked immunosorbent (ELISA) assays, respectively. The in vitro antioxidant property of the extract was also investigated by 1,1- diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) assays. Results: Levels of IL-6, NO, and ROS in LPS-treated BV2 cells were significantly higher than those in control (p < 0.01). However, exposure of LPS-treated BV2 cells to O. indicum extract led to a marked decrease in the levels of these parameters, when compared to the untreated cells (p < 0.01). Results from DPPH and ABTS assays showed that the O. indicum extract exhibited good antioxidant properties, with total flavonoid and total phenolic contents of 115.58 ± 1.09 and 131.04 ± 2.37 mg/g of dried extract, respectively. Conclusion: The results demonstrate that O. indicum fruit exerts anti-oxidant and anti-inflammatory effects in LPS-stimulated BV2 cells. Thus O. indicum fruit might be beneficial in the development of novel anti-oxidative and anti-neuroinflammatory herbal medicines. However, the mechanisms by which O. indium fruits reduces NO and IL-6 needs to be further investigated. [ABSTRACT FROM AUTHOR]

Introduction: A precise and reliable screening assay for glucose 6‐phosphate dehydrogenase (G6PD) deficiency would greatly help avoiding unwanted outcomes due to bilirubin neurotoxicity in neonatal jaundice and antimalarial‐induced haemolytic anaemia in malaria patients. Currently, available assays are laborious and require sophisticated laboratory expertise. This study aimed to evaluate the performance of a recently introduced automated screening assay for G6PD deficiency by comparing with a routine spectrophotometric assay. Methods: An automated UV‐based enzymatic (Mindray, PRC) and spectrophotometric assays were performed simultaneously in parallel to determine G6PD activity in 251 blood samples from the subjects. Results: The median G6PD activity value from spectrophotometric assay was significantly lower than that of from the automated assay. The mean difference was −2.0 U/g haemoglobin (−7.3 to 3.2; P < 0.0001). The mean activity values of both assays were strongly correlated with Pearson's correlation coefficient of r = 0.8. Cohen's kappa statistics between assays was 0.77 (0.70‐0.83). The sensitivity, specificity, positive and negative predictive values of the automated assay were 85.7%, 99.2%, 85.7%, 99.2%, respectively. The sensitivity and positive predictive values of the automated assay for identifying intermediate G6PD activity levels were 40.0% and 25.0%, respectively. Genotyping was performed to confirm G6PD deficient and intermediate samples. The turnaround time for 40 samples was 60 minutes for the automated assay and 300 minutes for spectrophotometric assay. Conclusion: The automated assay for the detection of G6PD deficiency is comparable to a routine spectrophotometric assay and help reducing sample handling time. However, the assay shows limitation in identifying individuals with G6PD intermediate. [ABSTRACT FROM AUTHOR]

Purpose: To determine the antioxidant and anti-inflammatory effects of Aegle marmelos fruit and Moringa oleifera (M. oleifera) Lam leaf extracts on LPS-stimulated BV2 microglial cells. Methods: BV2 cells were incubated with LPS for 24 h in the presence or absence of A. marmelos fruit extract or M. oleifera leaf extract. Subsequently, the levels of reactive oxygen species (ROS), nitric oxide (NO), tumor necrosis factor α (TNF α), and interleukins (IL) 6 were assessed. Thereafter, 2,2- diphenyl-1-picrylhydraz (DPPH), 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS), and ferric reducing antioxidant power (FRAP) antioxidant assays were performed. Results: The levels of ROS, NO, TNF α, and IL 6 in LPS-treated BV2 cells were significantly higher when compared to those in control group (p < 0.05). However, exposure of LPS-treated BV2 cells to either A. marmelos or M. oleifera leaf extract led to significant decrease in the levels of ROS and inflammatory mediators, when compared with LPS-treated group (p < 0.05). Decrease in the levels of ROS and inflammatory mediators in M. oleifera leaf extract-treated cells was also significantly larger when compared with A. marmelos-treated group (p < 0.05). The DPPH, ABTS, and FRAP assays revealed that M. oleifera leaf extracts had greater antioxidant capacity than A. marmelos extracts. Conclusion: Aegle marmelos fruit and M. oleifera leaf extracts exert antioxidant and anti-inflammatory effects on LPS activated BV2 cells in an in vitro model. Therefore, A. marmelos fruit and M. oleifera leaf extracts may be agents in the development of novel anti oxidative and anti-neuroinflammatory herbal medicines, or as food supplements. [ABSTRACT FROM AUTHOR]

Abnormal iron accumulation has been consistently reported in specific brain regions of many neurodegenerative diseases. At cellular level, iron is unusually observed in microglia, immune effector cell of the brain. Most evidence has provided that upon activation, microglia produces neurotoxins and different kinds of inflammatory mediators. Therefore, it is believed that activated microglia is actively involved in neurodegenerative process. Using a rat microglial cell line (HAPI), the present study was designed to address the role of iron for immune function of microglia, in particular, the production of Nitric Oxide (NO) in the presence or absence of estrogen, a potential neuroprotective agent. The present results demonstrated that exposure of microglia to iron significantly decreased lipopolysaccaride-induced NO production, as determined by nitrite accumulation in the cell culture medium, and such effect of iron was potentiated by increasing concentration of estrogen. Transcript analysis revealed that estrogen, but not iron, decreased the expression of inducible Nitric Oxide Synthase (iNOS). These results demonstrate that estrogen enhances the inhibitory effect of iron on microglial NO production by decreasing mRNA expression of iNOS and also suggest that iron sequestration by microglia under neuropathological conditions could be a protective mechanism against NO-induced neurotoxicity.