Green Synthesis of Gold Nanoparticles Using Liquiritin and Other Phenolics from Glycyrrhiza glabra and Their Anti-Inflammatory Activity.

Phenolic compounds are the main phytochemical constituents of many higher plants. They play an important role in synthesizing metal nanoparticles using green technology due to their ability to reduce metal salts and stabilize them through physical interaction/conjugation to the metal surface. Six pure phenolic compounds were isolated from licorice (Glycyrrhiza glabra) and employed in synthesizing gold nanoparticles (AuNPs). The isolated compounds were identified as liquiritin (1), isoliquiritin (2), neoisoliquiritin (3), isoliquiritin apioside (4), liquiritin apioside (5), and glabridin (6). The synthesized AuNPs were characterized using UV, zeta sizer, HRTEM, and IR and tested for their stability in different biological media. The phenolic isolates and their corresponding synthesized NP conjugates were tested for their potential in vitro cytotoxicity. The anti-inflammatory effects were investigated in both normal and inflammation-induced settings, where inflammatory biomarkers were stimulated using lipopolysaccharides (LPSs) in the RAW 264.7 macrophage cell line. LPS, functioning as a mitogen, promotes cell growth by reducing apoptosis, potentially contributing to observed outcomes. Results indicated that all six pure phenolic isolates inhibited cell proliferation. The AuNP conjugates of all the phenolic isolates, except liquiritin apioside (5), inhibited cell viability. LPS initiates inflammatory markers by binding to cell receptors and setting off a cascade of events leading to inflammation. All the pure phenolic isolates, except isoliquiritin, neoisoliquiritin, and isoliquiritin apioside inhibited the inflammatory activity of RAW cells in vitro.

The Effects of Endocrine Disrupting Chemicals on Biomarkers of Inflammation Produced by Lipopolysaccharide Stimulated RAW264.7 Macrophages.

Endocrine disrupting chemicals (EDCs) are common pollutants in the environment and can induce disruption of the endocrine and immune systems. The present study evaluated the effects of selected common environmental EDCs on secretion of inflammatory biomarkers by RAW264.7 cells. The EDCs investigated were Estradiol (E2), 5α-dihydrotestosterone (DHT), and Bisphenol A (BPA). To evaluate if the effects caused by EDCs were modulated by steroid hormone receptors, antagonists of estrogen and androgen receptors were used. The steroid receptor antagonists used were Tamoxifen, an estrogen receptor antagonist, and Flutamide, an androgen receptor antagonist. Secretion of biomarkers of inflammation, namely nitric oxide (NO) and interleukin 6 (IL-6), were monitored. The NO was determined using Griess reaction and IL-6 was measured by enzyme linked immunosorbent assay (ELISA). Although 5 µg/mL E2, DHT, and BPA were not toxic to RAW264.7 cell cultures, the same treatments significantly (p < 0.001) reduced both NO and IL-6 secretion by lipopolysaccharide (LPS)-stimulated RAW264.7 cell cultures. The suppression of NO and IL-6 secretion indicate inhibition of inflammation by DHT, E2, and BPA. The inhibitory effects of DHT, E2 and BPA are partially mediated via their cellular receptors, because the effects were reversed by their respective receptor antagonists. Flutamide reversed the effects of DHT, while Tamoxifen reversed the effects of E2 and BPA. In conclusion, E2, BPA, and DHT inhibit the synthesis of inflammation biomarkers by LPS-stimulated RAW264.7 cells. The inhibitory effects of EDCs can be partially reversed by the addition of an estrogen receptor antagonist for E2 and BPA, and an androgenic receptor antagonist for DHT. The inhibition of inflammatory response in stimulated RAW264.7 cells may be a useful bioassay model for monitoring estrogenic and androgenic pollutants.

Evaluation of cytotoxicity and inflammatory activity of wastewater collected from a textile factory before and after treatment by coagulation-flocculation methods.

Effective treatment of textile effluent prior to discharge is necessary in order to avert the associated adverse health impacts on human and aquatic life. In the present investigation, coagulation/flocculation processes were evaluated for the effectiveness of the individual treatment. Effectiveness of the treatment was evaluated based on the physicochemical characteristics. The quality of the pre-treated and post-flocculation treated effluent was further evaluated by determination of cytotoxicity and inflammatory activity using RAW264.7 cell cultures. Cytotoxicity was determined using WST-1 assay. Nitric oxide (NO) and interleukin 6 (IL-6) were used as biomarkers of inflammation. NO was determined in cell culture supernatant using the Griess reaction assay. The IL-6 secretion was determined using double antibody sandwich enzyme linked immunoassay (DAS ELISA). Cytotoxicity results show that raw effluent reduced the cell viability significantly (P < 0.001) compared to the negative control. All effluent samples treated by coagulation/flocculation processes at 1 in 100 dilutions had no cytotoxic effects on RAW264.7 cells. The results on inflammatory activities show that the raw effluent and effluent treated with 1.6 g/L of Fe-Mn oxide induced significantly (P < 0.001) higher NO production than the negative control. The inflammatory results further show that the raw effluent induced significantly (P < 0.001) higher production of IL-6 than the negative control. Among the coagulants/flocculants evaluated Al2(SO4)3.14H2O at a dosage of 1.6 g/L was the most effective to remove both toxic and inflammatory pollutants. In conclusion, the inflammatory responses in RAW264.7 cells can be used as sensitive biomarkers for monitoring the effectiveness of coagulation/flocculation processes used for textile effluent treatment.

The assessment of inflammatory activity and toxicity of treated sewage using RAW264.7 cells.

Toxicity and inflammatory activity of wastewater samples were evaluated using RAW264.7 cells as a bioassay model. The RAW264.7 cell cultures were exposed to sterile filtered wastewater samples collected from a sewage treatment plant. Cell viability was evaluated using WST-1 and XTT assays. Inflammatory effects of samples were assessed by determination of nitric oxide (NO) and interleukin 6 (IL-6). The NO was estimated using the Griess reaction and IL-6 was measured by enzyme-linked immunoassay. All samples had no toxicity effects to RAW264.7 cells, however they significantly (P < 0.001) induced NO and IL-6 production. The highest NO (12.5 ± 0.38 µM) and IL-6 (25383.84 ± 2327 pg/mL) production was induced by postbiofiltration sample. Final effluent induced the lowest inflammatory response, which indicates effective sewage treatment. In conclusion, wastewater samples can induce inflammatory activities in RAW264.7 cells. The RAW264.7 cells, therefore, can be used as a model for monitoring the quality of treated sewage.

Ecotoxicity of silver nanomaterials in the aquatic environment: a review of literature and gaps in nano-toxicological research.

There has been extensive growth in nanoscale technology in the last few decades to such a degree that nanomaterials (NMs) have become a constituent in a wide range of commercial and domestic products. With NMs already in use in several consumer products, concerns have emerged regarding their potential adverse environmental impacts. Although research has been undertaken in order to minimise the gaps in our understanding of NMs in the environment, little is known about their bioavailability and toxicity in the aquatic environment. Nano-toxicology is defined as the study of the toxicity of nanomaterials. Nano-toxicology studies remain poorly and unevenly distributed. To date most of the research undertaken has been restricted to a narrow range of test species such as daphnids. Crabs are bio-indicators that can be used for toxicological research on NMs since they occupy a significant position in the aquatic food chain. In addition, they are often used in conventional ecotoxicological studies due to their high sensitivity to environmental stressors and are abundantly available. Because they are benthic organisms they are prone to contaminant uptake and bioaccumulation. To our knowledge the crab has never been used in nano-toxicological studies. In this context, an extensive review on published scientific literature on the ecotoxicity of silver NPs (AgNPs) on aquatic organisms was conducted. Some of the most common biomarkers used in ecotoxicological studies are described. Emphasis is placed on the use of biomarker responses in crabs as monitoring tools, as well as on its limitations. Additionally, the gaps in nano-toxicological research and recommendations for future research initiatives are addressed.

In vitro interleukin-6 release in whole blood cultures in samples taken at rest from triathletes and professional rugby players.

The aim of this study was to investigate the endotoxin-induced interleukin-6 (IL-6) release in whole blood cultures from samples taken at rest, 24 h post-exercise, from a control group of recreationally trained individuals (C), a group of highly trained triathletes (TA) and a group of highly trained professional rugby players (RP). Fifteen RP [mean (SD): age 26 (3) years, height 1.90 (0.2) m, body mass 104.5 (12.2 kg)], 13 male TA [age 33 (5) years, height 1.78 (0.1) m, body mass 76.3 (12.6) kg] and eight recreationally active male volunteers [age 28 (6) years, height 1.80 (0.1) m, body mass 72.3 (7.3) kg] participated in the study. Plasma IL-6 concentration and in vitro IL-6 synthesis by whole blood cultures were measured in samples taken at rest. Plasma IL-6 concentration was significantly higher ( P<0.01) for the RP and TA groups than for C, as were the in vitro basal and endotoxin activated concentrations. However, after endotoxin stimulation, newly induced IL-6 concentration was significantly lower ( P<0.01) in the RP and TA than in the C group. Therefore, professional rugby players have a similar IL-6 release of whole blood cultures in vitro to that of triathletes. Specifically, mononuclear cells appear to be chronically activated to spontaneously release IL-6, but have a decreased capacity to respond to a further stimulus. Amongst possible explanations for this, the most likely is counter-regulation due to already elevated IL-6 release.