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 Stability and Anti-Angiogenic Properties of Titanium Dioxide Nanoparticles (TiO2NPs) Using Caco-2 Cells.

Titanium dioxide nanoparticles (TiO2NPs) are found in a wide range of products such as sunscreen, paints, toothpaste and cosmetics due to their white pigment and high refractive index. These wide-ranging applications could result in direct or indirect exposure of these NPs to humans and the environment. Accordingly, conflicting levels of toxicity has been associated with these NPs. Therefore, the risk associated with these reports and for TiO2NPs produced using varying methodologies should be measured. This study aimed to investigate the effects of various media on TiO2NP properties (hydrodynamic size and zeta potential) and the effects of TiO2NP exposure on human colorectal adenocarcinoma (Caco-2) epithelial cell viability, inflammatory and cell stress biomarkers and angiogenesis proteome profiles. The NPs increased in size over time in the various media, while zeta potentials were stable. TiO2NPs also induced cell stress biomarkers, which could be attributed to the NPs not being cytotoxic. Consequently, TiO2NP exposure had no effects on the level of inflammatory biomarkers produced by Caco-2. TiO2NPs expressed some anti-angiogenic properties when exposed to the no-observed-adverse-effect level and requires further in-depth investigation.

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.

The effects of silver nanoparticles on RAW 264.7. Macrophages and human whole blood cell cultures.

Silver nanoparticles (AgNPs) are commonly found in consumer products due to their antimicrobial properties. This study evaluated the effects of AgNPs on the murine macrophage cell line RAW 264.7 and human whole blood cell cultures (WBCs). Effects of AgNPs on RAW cells were assessed in the presence or absence of lipopolysaccharide (LPS). Effects of AgNPs on WBCs were monitored under basal conditions and in the presence of either LPS or phytohaemmagglutinin (PHA). AgNPs were cytotoxic to WBCs at 250 µg/ml. Under basal conditions, RAW cells ≥ 62.5. µg/ml and WBCs > 25 µg/ml AgNPs induced biomarkers associated with inflammation. Under LPS stimulated conditions, 250 µg/ml AgNP inhibited biomarkers associated with inflammation for both cultures. Under basal conditions, and in the presence of 250 µg/ml AgNP, WBCs produced acquired immune system cytokines IL-10 and IFNγ. IL-10 synthesis by WBCs was partially inhibited by 250 µg/ml AgNP in the presence of PHA. Proteome profiles of RAW cell supernatants show that AgNPs modulate biomarkers associated with inflammation. WBCs proteome analysis shows modulation of biomarkers associated with anti-inflammatory effects.

Quantitative assessment of heavy metal effects on sperm function using computer-aided sperm analysis and cytotoxicity assays.

One known environmental risk factor impacting on human reproduction is heavy metal pollution. Although some metals (e.g., Cu, Se and Zn) have protective effects on the male reproductive system in low doses, heavy metals can accumulate to toxic levels and result in poor semen quality and decreased sperm function. We investigated the effect of CuSO4 and CdCl2 (10, 50, 100 and 250 µg/ml or 500 µg/ml) on human sperm motility and vitality by using computer-aided sperm analysis (CASA) and two cytotoxicity assays (WST-1 and XTT). Several sperm motility parameters were significantly reduced after 5 hr of exposure to the highest concentrations of CuSO4 (250 µg/ml) and CdCl2 (500 µg/ml). The WST-1 assay also revealed significantly lower absorbance values for 50, 100 and 250 µg/ml CuSO4 and for 500 µg/ml CdCl2 ; however, no significant effect was seen with XTT. The calculated average IC50 value was 50.31±  4.34 µg/ml for CuSO4 and 392.32  ±76.79 µg/ml for CdCl2 . The effects of these metals were confirmed with MgCl2 , a positive control. This study provides threshold concentrations for the harmful effect of CuSO4 and CdCl2 on human spermatozoa and recommends the use of WST-1 as vitality assay in future in vitro studies.

The Effects of Carbon Dots on Immune System Biomarkers, Using the Murine Macrophage Cell Line RAW 264.7 and Human Whole Blood Cell Cultures.

Carbon dots (CDs) are engineered nanoparticles that are used in a number of bioapplications such as bioimaging, drug delivery and theranostics. The effects of CDs on the immune system have not been evaluated. The effects of CDs on the immune system were assessed by using RAW 264.7 cells and whole blood cell cultures. RAW cells were exposed to CD concentrations under basal conditions. Whole blood cell cultures were exposed to CD concentrations under basal conditions or in the presence of the mitogens, lipopolysaccharide (LPS) or phytohaemmagglutinin (PHA). After exposure, a number of parameters were assessed, such as cell viability, biomarkers of inflammation, cytokine biomarkers of the acquired immune system and a proteome profile analysis. CDs were cytotoxic to RAW and whole blood cell cultures at 62.5, 250 and 500 µg/mL, respectively. Biomarkers associated with inflammation were induced by CD concentrations ≥250 and 500 µg/mL under basal conditions for both RAW and whole blood cell cultures, respectively. The humoral immune cytokine interleukin (IL)-10 was increased at 500 µg/mL CD under both basal and PHA activated whole blood cell culture conditions. Proteome analysis supported the inflammatory data as upregulated proteins identified are associated with inflammation. The upregulated proteins provide potential biomarkers of risk that can be assessed upon CD exposure.

Effects of Graphene Oxide Nanoparticles on the Immune System Biomarkers Produced by RAW 264.7 and Human Whole Blood Cell Cultures.

Graphene oxide nanoparticles (GONPs) have attracted a lot of attention due to their many applications. These applications include batteries, super capacitors, drug delivery and biosensing. However, few studies have investigated the effects of these nanoparticles on the immune system. In this study, the in vitro effects of GONPs on the immune system was evaluated by exposing murine macrophages, RAW 264.7 cells and human whole blood cell cultures (to GONPs. The effects of GONPs on RAW cells were monitored under basal conditions. The whole blood cell cultures were exposed to GONPs in the presence or absence of the mitogens lipopolysaccharide (LPS) and phytohaemmagglutinin (PHA). A number of parameters were monitored for both RAW and whole blood cell cultures, these included cytotoxicity, inflammatory biomarkers, cytokines of the acquired immune system and a proteome profile analysis. The GONPs were cytotoxic to both RAW and whole blood cell cultures at 500 µg/mL. In the absence of LPS, GONPs elicited an inflammatory response from the murine macrophage, RAW and whole blood cell cultures at 15.6 and 5 µg/mL respectively. This activation was further corroborated by proteome profile analysis of both experimental cultures. GONPs inhibited LPS induced interleukin 6 (IL-6) synthesis and PHA induced interferon gamma (IFNγ) synthesis by whole blood cell cultures in a dose dependent manner. In the absence of mitogens, GONPs stimulated IL-10 synthesis by whole blood cell cultures. The current study shows that GONPs modulate immune system biomarkers and that these may pose a health risk to individuals exposed to this type of nanoparticle.

The effect of sugar cane molasses on the immune and male reproductive systems using in vitro and in vivo methods.

OBJECTIVES: Sugar cane molasses is a commonly used ingredient in several food products. Contrasting reports suggest that molasses may have potential adverse or beneficial effects on human health. However, little evidence exists that examines the effects of molasses on the different physiological systems. This study investigated the effects of sugar cane molasses on various physiological systems using in vivo and in vitro methods. MATERIALS AND METHODS: Molasses was administered orally to BALB/c, male mice and animals were randomly assigned into either a treatment or control group. General physiological changes, body weight and molasses intake of animals were monitored. At the end of the exposure period, collected blood samples were evaluated for potential toxicity using plasma biomarkers and liver enzyme activity. Immunised treated and untreated mice were evaluated for antibody titre to determine the effect of molasses on the immune response. To investigate the impact of molasses on testicular steroidogenesis, testes from both treated and control groups were harvested, cultured and assayed for testosterone synthesis. RESULTS: Findings suggest that fluid intake by molasses-treated animals was significantly increased and these animals showed symptoms of loose faeces. Molasses had no significant effect on body weight, serum biomarkers or liver enzyme activity (P>0.05). Immunoglobulin-gamma anti-antigen levels were significantly suppressed in molasses-treated groups (P=0.004). Animals subjected to molasses exposure also exhibited elevated levels of testosterone synthesis (P=0.001). CONCLUSION: Findings suggests that molasses adversely affects the humoral immune response. The results also promote the use of molasses as a supplement to increase testosterone levels.

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.

Effect of temperature on oxidative stress parameters and enzyme activity in tissues of Cape River crab (Potamanautes perlatus) following exposure to silver nanoparticles (AgNP).

Biomarkers of oxidative stress have been widely used in environmental assessments to evaluate the effects of exposure of aquatic organisms to contaminants from various anthropogenic sources. Silver nanoparticles (AgNP), the most produced NP worldwide and used in several consumer products, are known to produce oxidative stress in aquatic organisms. Similarly, temperature is also known to affect reactive oxygen species (ROS) by influencing the inputs of contaminants into the environment, as well as altering behavior, fate, and transport. Aquatic ecosystems are affected by both anthropogenic releases of contaminants and increased temperature. To test this hypothesis, the influence of AgNP and temperature in the response to multiple biomarkers of oxidative stress was studied in the gills and hepatopancreas of the Cape River crab Potamonautes perlatus. Responses were assessed through activities of antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), and the nonenzymatic antioxidant glutathione S-transferase (GST). The response of the oxidative stress biomarkers analyzed was always higher in hepatopancreas than in gills. Elevated temperatures (28°C) induced oxidative stress by increasing SOD, CAT, and GST activities, particularly at 100 µg/ml AgNP. These data indicate that AgNP-mediated toxicity to P. perlatus is modulated by elevated temperatures, but this relationship is not linear. Co-effects of AgNP and temperature are reported for the first time in P. perlatus.

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.

The effectiveness of sewage treatment processes to remove faecal pathogens and antibiotic residues.

Pathogens and antibiotics enter the aquatic environment via sewage effluents and may pose a health risk to wild life and humans. The aim of this study was to determine the levels of faecal bacteria, and selected antibiotic residues in raw wastewater and treated sewage effluents from three different sewage treatment plants in the Western Cape, South Africa. Sewage treatment plant 1 and 2 use older technologies, while sewage treatment plant 3 has been upgraded and membrane technologies were incorporated in the treatment processes. Coliforms and Escherichia coli (E. coli) were used as bioindicators for faecal bacteria. A chromogenic test was used to screen for coliforms and E. coli. Fluoroquinolones and sulfamethoxazole are commonly used antibiotics and were selected to monitor the efficiency of sewage treatment processes for antibiotic removal. Enzyme Linked Immunosorbent Assays (ELISAs) were used to quantitate antibiotic residues in raw and treated sewage. Raw intake water at all treatment plants contained total coliforms and E. coli. High removal of E. coli by treatment processes was evident for treatment plant 2 and 3 only. Fluoroquinolones and sulfamethoxazole were detected in raw wastewater from all sewage treatment plants. Treatment processes at plant 1 did not reduce the fluoroquinolone concentration in treated sewage effluents. Treatment processes at plant 2 and 3 reduced the fluoroquinolone concentration by 21% and 31%, respectively. Treatment processes at plant 1 did not reduce the sulfamethoxazole concentration in treated sewage effluents. Treatment processes at plant 2 and 3 reduced sulfamethoxazole by 34% and 56%, respectively. This study showed that bacteria and antibiotic residues are still discharged into the environment. Further research needs to be undertaken to improve sewage treatment technologies, thereby producing a better quality treated sewage effluent.

The implementation of a battery of in vivo and in vitro bioassays to assess river water for estrogenic endocrine disrupting chemicals.

Previous research has shown that accurate evaluation of environmental water samples for estrogenic activity requires a panel of in vitro and in vivo bioassays, which are based on different molecular and cellular action mechanisms. In the current study, a test battery containing four assays was used to analyze water from the Eerste River, South Africa for estrogenicity. Three sites were used for analysis, namely Jonkershoek (control site situated in the mountains at the origin of the Eerste River), sewage effluent from Stellenbosch sewage treatment works and Spier site (sampling site on the Eerste River downstream from Stellenbosch). Estrogenicity was determined using an estrone enzyme linked immuno sorbent assay (ELISA), estrogen induced proliferation of human breast cancer adenocarcinoma cells (MCF-7) also known as the E-SCREEN, estrogen induced suppression of estrogen receptor alpha protein expression (ER-α) in MCF-7 cells (ERα assay) and by monitoring estrogen induced vitellogenin (VTG) synthesis in juvenile Oreochromis mossambicus (VTG assay). Low concentrations of estrone (ranging between 1.4 and 2.2 ng/l) near the detection limit of the assay were detected in samples collected from Jonkershoek. Water from this site shows no estrogenicity in the E-SCREEN, ERα assay or VTG synthesis bioassay. The estrone concentrations in the sewage effluent extracts, as well as Spier site extracts, ranged between 14.7 and 19.4 ng/l. The assays using ERα induction by the MCF-7 cell line, MCF-7 proliferation and in vivo VTG synthesis by juvenile tilapia showed that these samples are estrogenic. The results obtained for the assays in the battery are comparable.

The effect of Tulbaghia violacea extracts on testosterone secretion by testicular cell cultures.

AIM OF THE STUDY: This study aimed to determine the effect of Tulbaghia violacea Harv. on the male reproductive system in vitro by using testicular cell cultures. Tulbaghia violacea is a plant species indigenous to southern Africa and is used locally as a herbal remedy/medicine to treat several ailments. MATERIALS AND METHODS: A 50% ethanol extract of Tulbaghia violacea was prepared. Three-month old male Balb/C mice were sacrificed and testicular cell cultures were prepared. Cells were then treated with varying concentrations of the Tulbaghia violacea ethanol extract (with/without Luteinizing hormone (LH)-treatment) and incubated for 4 h. Hormone production and cell viability were evaluated. RESULTS: Treatment of cells with Tulbaghia violacea (312.5-5000 µg ml(-1)) significantly increased (P<0.05) LH-induced testosterone production as compared to vehicle-treated control (DMSO) whereas cells without LH-treatment showed no significant change in testosterone concentrations. No significant effect on cell viability was observed at all concentrations tested. CONCLUSIONS: The data presented shows that Tulbaghia violacea has androgenic properties. Further studies are warranted to determine and clarify the exact mechanisms involved.

The effects of Saccharum officinarium (sugar cane) molasses on cytokine secretion by human blood cultures.

This study investigated the effects of sugar cane molasses on the immune system, using cytokines as biomarkers. Whole blood cultures, stimulated in vitro with endotoxin or PHA, were incubated with various concentrations of molasses. No cell death occurred in whole blood cultures incubated with molasses samples. The addition of molasses (800 microg/mL) to unstimulated whole blood cultures resulted in increased levels of the biomarker of inflammation, Interleukin-6 (P < 0.001) and also the biomarker of humoral immunity, Interleukin-10 (P < 0.001). Molasses addition (800 microg/mL) to unstimulated whole blood cultures has no effect on the cell mediated immunity biomarker, Interferon gamma secretion. Molasses has no effect on Interleukin-6, Interleukin-10 and Interferon gamma secretion in stimulated whole blood cultures. Immunostimulation by molasses requires further investigation as it may have potential health impacts.

The in vitro effects of Rooibos and Black tea on immune pathways.

The in vitro effects of Aspalathus linearis (Rooibos tea) and Camellia sinensis (Black tea) on biomarkers of specific immune pathways were determined using whole blood culture assays. Stimulated and unstimulated whole blood cultures were incubated with tea extracts. Enzyme linked immunosorbent assays were used to screen spent culture medium for Interleukin-6, Interleukin-10 and Interferon gamma as biomarkers for inflammation, humoral immunity, and cell mediated immunity, respectively. Rooibos and Black tea addition to unstimulated whole blood cultures induced higher Interleukin-6, Interleukin-10, and Interferon gamma secretion. Addition of Rooibos tea to stimulated whole blood cultures induced higher Interleukin-6, lower Interleukin-10, and had no effect on Interferon gamma secretion. Black tea addition to stimulated whole blood cultures inhibited Interleukin-6, Interleukin-10, and Interferon gamma production. The data indicates that Rooibos and Black tea modulates immune function in vitro.

Rapid detection of selected steroid hormones from sewage effluents using an ELISA in the Kuils River water catchment area, South Africa.

Steroid hormones are naturally synthesized by both humans and animals and are released into the environment. Significant levels of steroid hormones have been detected in sewage effluent around the world. The potential problem is that these hormones may interfere with the normal function of the endocrine systems, thus affecting reproduction and development in wildlife. Due to the major shortage of water in Western Cape, South Africa there is a great need to recycle water by either direct or indirect methods. The treated sewage effluent-natural surface water mixture found in the Kuils and Eerste Rivers is used directly for irrigation of agricultural areas. Sewage effluents were collected from four sites (Jonkershoek, Belville, Zandvliet, and Macassar) and subjected to C(18) solid phase extraction. Commercially available rapid ELISA kits were validated for the quantification of estrogens in these sewage effluent samples. Analysis of estrone, estradiol, and estriol levels showed a significant difference between the control site (Jonkershoek) and sewage effluent from the three sewage treatment works. Steroid hormone concentrations detected in these sewage effluents were similar to reports from Britain, Italy, Germany, Canada, and Netherlands.

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.