Molecular interactions and signal transfer between sensory rhodopsin-I and its cognate transducer

SRI is unique among known photoreceptors in that it produces opposite signals depending on the color of light stimuli. Absorption of orange light (587 nm) triggers an attractant response by the cell, whereas absorption of orange light followed by near-UV light (373 run) triggers a repellent response. Using behavioral mutants that exhibit aberrant color-sensing ability, we tested a two-conformation equilibrium model, using FRET and EPR spectroscopy. The essence of the model applied to SRI-HtrI is that the complex exists in a metastable two-conformer equilibrium which is shifted in one direction by orange light absorption (producing an attractant signal) and in the opposite direction by a second UV-violet photon (producing a repellent signal). First, by FRET we found that the E-F cytoplasmic loop of SRI moves toward the RAMP domain of the HtrI transducer during the formation of the orange-light activated signaling state of the complex. This is the first localization of a change in the physical relationship between the receptor and transducer subunits of the complex and provides a structural property of the two proposed conformers that we can monitor. Second, EPR spectra of a spin label probe at this cytoplasmic position showed shifts in the dark in the mutants toward shorter or longer EF loop-RAMP distances, explaining their behavior in terms of their mutations causing pre-stimulus shifts into one or the other conformer. ^ Next, we applied a novel electrophysiological method for monitoring the directionality of proton movement during photoactivation of SRI, to investigate the process of proton transfer in the photoactive site from the chromophore to proton acceptors on both the wildtype and aberrant color-response mutants. We observed an unexpected and critical difference in the two signaling conformations of the SRI-HtrI complex. The finding is that the vectoriality (i.e. movement away or toward the cytoplasm) of the light-induced proton transfer from the chromophore to the protein is opposite in formation of the two conformations. Retinylidene proton transfer is a common critical process in rhodopsins and these results are the first to show differences in vectoriality in a rhodopsin receptor, and to demonstrate functional importance of the direction of proton transfer. ^

Emergency department usage patterns among residents of Hardin, Jefferson, and Orange Counties in southeast Texas

Emergency departments (EDs) have been called the net below the safety net due to their long history of providing care to the uninsured and others lacking access to the healthcare system. In past years, those with Medicaid and, more recently, those with Medicare, are also utilizing the ED as a medical home for routine primary care. There are many reasons for this but the costs to the community have become increasingly burdensome. ^ To evaluate how often the ED is being utilized for primary care, we applied a standardized tool, the New York University Algorithm, to over 43,000 ED visits when no hospitalization was required made by Hardin, Jefferson, and Orange County residents over a 12 month period. We compared our results to Harris County, where studies using the same framework have been performed, and found that sizeable segments of the population in both areas are utilizing the ED for non-emergent primary care that could be treated in a more cost-effective community setting. ^ We also analyzed our dataset for visit-specific characteristics. We found evidence of two possible health care disparities: (1) Blacks had a higher rate of primary care-related ED visits in relation to their percentage of the population when compared to other racial/ethnic groups; and (2) when form of payment is considered, the uninsured were more likely to have a primary care-related ED visit than any other group. These findings suggest a lack of community-based primary care services for the medically needy in Southeast Texas. ^ We believe that studies such as this are warranted elsewhere in Texas as well. We plan to present our findings to local policy makers, who should find this information helpful in identifying gaps in the safety net and assist them in better allocating scarce community resources. ^