The sodium iodide symporter (NIS) mediates iodide (I−) transport in the thyroid gland and other tissues and is of increasing importance as a therapeutic target and nuclear imaging reporter. NIS activity in vitro is currently measured with radiotracers and electrophysiological techniques. We report on the development of a novel live cell imaging assay of NIS activity using the I−-sensitive and genetically encodable yellow fluorescent protein (YFP) variant YFP-H148Q/I152L. In FRTL-5 thyrocytes stably expressing YFP-H148Q/I152L, I− induced a rapid and reversible decrease in cellular fluorescence characterized by 1) high affinity for extracellular I− (35 μM), 2) inhibition by the NIS inhibitor perchlorate, 3) extracellular Na+ dependence, and 4) TSH dependence, suggesting that fluorescence changes are due to I− influx via NIS. Individual cells within a population of FRTL-5 cells exhibited a 3.5-fold variation in the rate of NIS-mediated I− influx, illustrating the utility of YFP-H148Q/I152L to detect cell-to-cell difference in NIS activity. I− also caused a perchlorate-sensitive decrease in YFP-H148Q/I152L fluorescence in COS-7 cells expressing NIS but not in cells lacking NIS. These results demonstrate that YFP-H148Q/I152L is a sensitive biosensor of NIS-mediated I− uptake in thyroid cells and in nonthyroidal cells following gene transfer and suggest that fluorescence detection of cellular I− may be a useful tool by which to study the pathophysiology and pharmacology of NIS.
Giant reed is a non-food, tall, rhizomatous, spontaneous perennial grass that is widely diffused in warm-temperate environments under different pedo-climatic conditions. In such environments, it is considered one of the most promising energy crops in terms of economic and environmental sustainability, as it can also be cultivated on marginal lands. Owing to its complex and recalcitrant structure due to the lignin content, the use of giant reed as a feedstock for biogas production is limited. Thus, pre-treatment is necessary to improve the methane yield. The objective of this review was to critically present the possible pre-treatment methods to allow the giant reed to be transformed in biogas. Among the studied pre-treatments (i.e., hydrothermal, chemical, and biological), alkaline pre-treatments demonstrated better effectiveness in improving the methane yield. A further opportunity is represented by hybrid pre-treatments (i.e., chemical and enzymatic) to make giant reed biomass suitable for bio-hydrogen production. So far, the studies have been carried out at a laboratory scale; a future challenge to research is to scale up the pre-treatment process to a pilot scale.
1 January 2015 in Environmental Engineering and Management Journal