Fluorescent proteins have been widely used in biology because of their compatibility and varied applications in living specimens. Fluorescent proteins are often undesirably sensitive to intracellular conditions such as pH and ion concentration, generating considerable issues at times. However, harnessing these intrinsic sensitivities can help develop functional probes. In this study, we found that the fluorescence of yellow fluorescent protein (YFP) depends on the protein concentration in the solution and that this dependence can be enhanced by adding a glycine residue in to the YFP; we applied this finding to construct an intracellular protein-crowding sensor. A Förster resonance energy transfer (FRET) pair, involving a cyan fluorescent protein (CFP) insensitive to protein concentration and a glycine-inserted YFP, works as a genetically encoded probe to evaluate intracellular crowding. By measuring the fluorescence of the present FRET probe, we were able to detect dynamic changes in protein crowding in living cells.

Abstract. Observations from the new Japanese geostationary satellite Himawari-8 permit quasi-real-time estimation of global shortwave radiation at an unprecedented temporal resolution. However, accurate comparisons with groundtruthing observations are essential to assess their uncertainty. In this study, we evaluated the Himawari-8 global radiation product AMATERASS using observations recorded at four SKYNET stations in Japan and, for certain analyses, from the surface network of the Japanese Meteorological Agency in 2016. We found that the spatiotemporal variability of the satellite estimates was smaller than that of the ground observations; variability decreased with increases in the time step and spatial domain. Cloud variability was the main source of uncertainty in the satellite radiation estimates, followed by direct effects caused by aerosols and bright albedo. Under all-sky conditions, good agreement was found between satellite and ground-based data, with a mean bias in the range of 20-30 W m −2 (i.e., AMATERASS overestimated ground observations) and a root mean square error (RMSE) of approximately 70-80 W m −2 . However, results depended on the time step used in the validation exercise, on the spatial domain, and on the different climatological regions. In particular, the validation performed at 2.5 min showed largest deviations and RMSE values ranging from about 110 W m −2 for the mainland to a maximum of 150 W m −2 in the subtropical region. We also detected a limited overestimation in the number of clear-sky episodes, particularly at the pixel level. Overall, satellite-based estimates were higher under overcast conditions, whereas frequent episodes of cloud-induced enhanced surface radiation (i.e., measured radiation was greater than expected clear-sky radiation) tended to reduce this difference. Finally, the total mean bias was approximately 10-15 W m −2 under clear-sky conditions, mainly because of overall instantaneous direct aerosol forcing efficiency in the range of 120-150 W m −2 per unit of aerosol optical depth (AOD). A seasonal anticorrelation between AOD and global radiation differences was evident at all stations and was also observed within the diurnal cycle.

Abstract.Observations from the new Japanese geostationary satellite Himawari-8 permit quasi-real-time estimation of global shortwave radiation at an unprecedented temporal resolution. However, accurate comparisons with ground truthing observations are essential to assess their uncertainty. In this study, we evaluated the Himawari-8 global radiation product AMATERASS using observations recorded at four SKYNET stations in Japan and, for certain analyses, from the surface 15 network of the Japanese Meteorological Agency in 2016. We found that the spatiotemporal variability of the satellite estimates was smaller than that of the ground observations; variability decreased with increases in the time step and spatial domain. Cloud variability was the main source of uncertainty in the satellite radiation estimates, followed by direct effects caused by aerosols and bright albedo. Under all-sky conditions, good agreement was found between satellite and groundbased data, with a mean bias in the range of 20-30 W/m 2 (i.e., AMATERASS overestimated ground observations) and a root 20 mean square error of approximately 80 W/m 2 . However, results depended on the time step used in the validation exercise and on the spatial domain. We also detected a limited overestimation in the number of clear-sky episodes, particularly at the pixel level. Overall, satellite-based estimates were higher under overcast conditions, whereas frequent episodes of cloudinduced enhanced surface radiation (i.e., measured radiation was greater than expected clear-sky radiation) tended to reduce this difference. Finally, the total mean bias was reduced to approximately 10-15 W/m 2 under clear-sky conditions, mainly 25 because of overall instantaneous direct aerosol forcing efficiency in the range of 120-150 W/m 2 per unit of aerosol optical depth (AOD). A seasonal anti-correlation between AOD and global radiation differences was evident at all stations and was also observed within the diurnal cycle.

The glucocorticoid receptor (GR) is a transcription factor, which interacts with DNA and other cofactors to regulate gene transcription. Binding to other partners in the cell nucleus alters the diffusion properties of GR. Raster image correlation spectroscopy (RICS) was applied to quantitatively characterize the diffusion properties of EGFP labeled human GR (EGFP-hGR) and its mutants in the cell nucleus. RICS is an image correlation technique that evaluates the spatial distribution of the diffusion coefficient as a diffusion map. Interestingly, we observed that the averaged diffusion coefficient of EGFP-hGR strongly and negatively correlated with its transcriptional activities in comparison to that of EGFP-hGR wild type and mutants with various transcriptional activities. This result suggests that the decreasing of the diffusion coefficient of hGR was reflected in the high-affinity binding to DNA. Moreover, the hyper-phosphorylation of hGR can enhance the transcriptional activity by reduction of the interaction between the hGR and the nuclear corepressors.