Assessing the seasonal influence of sewage and agricultural nutrient inputs in a subtropical river estuary

A combination of physical and chemical measurements and biological indicators identified nutrient impacts throughout an Australian subtropical river estuary. This was a balance of sewage inputs in the lower river and agricultural inputs in the mid-upper river, the combined influence being greater in the wet season due to greater agricultural surface runoff. Field sampling in the region was conducted at 6 sites within the river, over 5 surveys to encapsulate both wet and dry seasonal effects. Parameters assessed were tissue nitrogen (N) contents and delta(15)N signatures of mangroves and macroalgae, phytoplankton nutrient addition bioassays, and standard physical and chemical variables. Strong spatial (within river) and temporal (seasonal) variability was observed in all parameters. Poorest water quality was detected in the middle (agricultural) region of the river in the wet season, attributable to large diffuse inputs in this region. Water quality towards the river mouth remained constant irrespective of season due to strong oceanic flushing. Mangrove and macroalgal tissue delta(15)N and %N proved a successful combination for discerning sewage and agricultural inputs. Elevated delta(15)N and %N represented sewage inputs, whereas low delta(15)N and elevated %N was indicative of agricultural inputs. Phytoplankton bioassays found the system to be primarily responsive to nutrient additions in the warmer wet season, with negligible responses observed in the cooler dry season. These results indicate that the Tweed River is sensitive to the different anthropogenic activities in its catchment and that each activity has a unique influence on receiving water quality.

Ratiometric and nonratiometric Ca2+ indicators for the assessment of intracellular free Ca2+ in a breast cancer cell line using a fluorescence microplate reader

Transporters of Ca2+ are potential drug targets and Ca2+ is a useful signal in the assessment of G-protein-coupled receptor activation. Assays involving the assessment of intracellular Ca2+ using microplate readers most often use Ca2+ indicators which do not exhibit a spectra shift on Ca2+ binding (e.g. fluo-3). Indicators that do exhibit a spectral shift upon Ca2+ binding (e.g. fura-2) offer potential advantages for the calibration of intracellular Ca2+ levels. However, experimental limitations may limit the use of ratiometric dyes in microplate readers capable of screening. In this study, we compared the assessment of intracellular Ca2+ in adherent breast cancer cells using ratiometric and nonratiometric Ca2+ indicators. Our results demonstrate that both fluo-3 and fura-2 detect ATP dose-dependent increases in intracellular Ca2+ in the MCF-7 breast cancer cell line and that some of the limitations in the use of fura-2 appear to be overcome by the use of glass bottom microplates. The calibrated intracellular Ca2+ levels derived using fura-2 are consistent with those from microscopy and cuvette-based studies. Fura-2 may be useful in microplate studies, where cell lines with different properties are compared or where screening treatments lead to differences in the number of cells or dye loading. (C) 2003 Elsevier B.V. All rights reserved.