Phosphocholine – an agonist of metabotropic but not of ionotropic functions of alpha9-containing nicotinic acetylcholine receptors

We demonstrated previously that phosphocholine and phosphocholine-modified macromolecules efficiently inhibit ATP-dependent release of interleukin-1beta from human and murine monocytes by a mechanism involving nicotinic acetylcholine receptors (nAChR). Interleukin-1beta is a potent pro-inflammatory cytokine of innate immunity that plays pivotal roles in host defence. Control of interleukin-1beta release is vital as excessively high systemic levels cause life threatening inflammatory diseases. In spite of its structural similarity to acetylcholine, there are no other reports on interactions of phosphocholine with nAChR. In this study, we demonstrate that phosphocholine inhibits ion-channel function of ATP receptor P2X7 in monocytic cells via nAChR containing alpha9 and alpha10 subunits. In stark contrast to choline, phosphocholine does not evoke ion current responses in Xenopus laevis oocytes, which heterologously express functional homomeric nAChR composed of alpha9 subunits or heteromeric receptors containing alpha9 and alpha10 subunits. Preincubation of these oocytes with phosphocholine, however, attenuated choline-induced ion current changes, suggesting that phosphocholine may act as a silent agonist. We conclude that phophocholine activates immuno-modulatory nAChR expressed by monocytes but does not stimulate canonical ionotropic receptor functions

Project ELLA: English Language and Literacy Acquisition

An address at the Annual Meeting of the American Educational Research Association, San Francisco, CA, April 9, 200

BLUF Domain Function Does Not Require a Metastable Radical Intermediate State

BLUF (blue light using flavin) domain proteins are an important family of blue light-sensing proteins which control a wide variety of functions in cells. The primary light-activated step in the BLUF domain is not yet established. A number of experimental and theoretical studies points to a role for photoinduced electron transfer (PET) between a highly conserved tyrosine and the flavin chromophore to form a radical intermediate state. Here we investigate the role of PET in three different BLUF proteins, using ultrafast broadband transient infrared spectroscopy. We characterize and identify infrared active marker modes for excited and ground state species and use them to record photochemical dynamics in the proteins. We also generate mutants which unambiguously show PET and, through isotope labeling of the protein and the chromophore, are able to assign modes characteristic of both flavin and protein radical states. We find that these radical intermediates are not observed in two of the three BLUF domains studied, casting doubt on the importance of the formation of a population of radical intermediates in the BLUF photocycle. Further, unnatural amino acid mutagenesis is used to replace the conserved tyrosine with fluorotyrosines, thus modifying the driving force for the proposed electron transfer reaction; the rate changes observed are also not consistent with a PET mechanism. Thus, while intermediates of PET reactions can be observed in BLUF proteins they are not correlated with photoactivity, suggesting that radical intermediates are not central to their operation. Alternative nonradical pathways including a keto–enol tautomerization induced by electronic excitation of the flavin ring are considered

Responses to LBNP in men with varying profiles of strength and aerobic capacity: Implications for flight crews

Hemodynamic and hormonal responses to lower-body negative pressure (LBNP) were examined in 24 healthy men to test the hypothesis that responsiveness of reflex control of blood pressure during orthostatic stress is associated with strength and/or aerobic capacity. Subjects underwent treadmill tests to determine peak oxygen uptake (peak VO2) and isokinetic dynamo meter tests to determine leg strength. Based on predetermined criteria, the subjects were classified into one of four fitness profiles of six subjects each matched for age, height, and weight: (1) low strength/low aerobic fitness; (2) low strength/high aerobic fitness; (3) high strength/low aerobic fitness; and (4) high strength/high aerobic fitness. Following 90 min of 6 degree head-down tilt (HDT), each subject underwent graded LBNP through -50 mmHg or presyncope, with maximal duration 15 min. All groups exhibited typical hemodynamic, hormonal, and fluid shift responses during LBNP, with no intergroup differences except for catecholamines. Seven subjects, distributed among the four fitness profiles, became presyncopal. Subjects who showed greatest reduction in mean arterial pressure (MAP) during LBNP had greater elevations in vasopressin and lesser increases in heart rate and peripheral resistance. Peak VO2 nor leg strength were correlated with fall in MAP or with syncopal episodes. We conclude that neither aerobic nor strength fitness characteristics are good predictors of responses to LBNP stress

The development and evaluation of an online application to assist in the extraction of data from graphs for use in systematic reviews

These are the data we generated in our evaluation of the graphical user interface. Please see our publication on Wellcome Open Research for information about the evaluations.These are the data we generated in our evaluation of the graphical user interface. Please see our publication on Wellcome Open Research for information about the evaluations