Browsing by Author "Pool, Edmund"
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Item The effects of carbon dots on immune system biomarkers, using the Murine Macrophage cell line RAW 264.7 and human whole blood cell cultures(MDPI, 2018) Lategan, Kim; Fowler, Jodi; Bayati, Mohamed; Fidalgo de Cortalezzi, Maria; Pool, EdmundAbstract: 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 RAW264.7 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.Item Quantitative assessment of heavy metal effects on sperm function using computer‐aided sperm analysis and cytotoxicity assays(Wiley, 2018) Hardneck, Farren; Israel, Gadieja; Pool, Edmund; Maree, LianaOne 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.