Slate Literary Magazine, 2012-2013

Trinity College\u27s Literary Magazine - student works.https://digitalrepository.trincoll.edu/slate/1000/thumbnail.jp

Preferential Phosphorylation of R-domain Serine 768 Dampens Activation of CFTR Channels by PKA

CFTR (cystic fibrosis transmembrane conductance regulator), the protein whose dysfunction causes cystic fibrosis, is a chloride ion channel whose gating is controlled by interactions of MgATP with CFTR's two cytoplasmic nucleotide binding domains, but only after several serines in CFTR's regulatory (R) domain have been phosphorylated by cAMP-dependent protein kinase (PKA). Whereas eight R-domain serines have previously been shown to be phosphorylated in purified CFTR, it is not known how individual phosphoserines regulate channel gating, although two of them, at positions 737 and 768, have been suggested to be inhibitory. Here we show, using mass spectrometric analysis, that Ser 768 is the first site phosphorylated in purified R-domain protein, and that it and five other R-domain sites are already phosphorylated in resting Xenopus oocytes expressing wild-type (WT) human epithelial CFTR. The WT channels have lower activity than S768A channels (with Ser 768 mutated to Ala) in resting oocytes, confirming the inhibitory influence of phosphoserine 768. In excised patches exposed to a range of PKA concentrations, the open probability (Po) of mutant S768A channels exceeded that of WT CFTR channels at all [PKA], and the half-maximally activating [PKA] for WT channels was twice that for S768A channels. As the open burst duration of S768A CFTR channels was almost double that of WT channels, at both low (55 nM) and high (550 nM) [PKA], we conclude that the principal mechanism by which phosphoserine 768 inhibits WT CFTR is by hastening the termination of open channel bursts. The right-shifted Po-[PKA] curve of WT channels might explain their slower activation, compared with S768A channels, at low [PKA]. The finding that phosphorylation kinetics of WT or S768A R-domain peptides were similar provides no support for an alternative explanation, that early phosphorylation of Ser 768 in WT CFTR might also impair subsequent phosphorylation of stimulatory R-domain serines. The observed reduced sensitivity to activation by [PKA] imparted by Ser 768 might serve to ensure activation of WT CFTR by strong stimuli while dampening responses to weak signals

An investigation of the stability and diffusivity of flexible lipid vesicles for transdermal insulin delivery

Gemstone Team No More NeedlesFlexible lipid vesicles have the potential of complementing or even replacing traditional needle injection methods for insulin delivery. Vesicles are made flexible by the incorporation of a chemical surfactant which may also hinder their stability. We studied the changes in the size and apparent flexibility of vesicles with varying surfactant concentrations over time and the effects these changes have on vesicle diffusion. We found that increased surfactant concentrations lead to greater size fluctuations. In addition, we witnessed a significant decrease in the flexibility of vesicles over six weeks, while the diffusivity of surfactant infused liposomes increased over a single week. Our data suggests that while surfactants are necessary in vesicles for transdermal drug delivery, their long-term stability is uncertain. Using our diffusion data, we developed a model to estimate the insulin delivering capacity of a hypothetical insulin patch which has the potential to stabilize vesicles for extended periods of time

Diabetes and cardiovascular disease: A statement for healthcare professionals from the American Heart Association

This statement examines the cardiovascular complications of diabetes mellitus and considers opportunities for their prevention. These complications include coronary heart disease (CHD), stroke, peripheral arterial disease, nephropathy, retinopathy, and possibly neuropathy and cardiomyopathy. Because of the aging of the population and an increasing prevalence of obesity and sedentary life habits in the United States, the prevalence of diabetes is increasing. Thus, diabetes must take its place alongside the other major risk factors as important causes of cardiovascular disease (CVD). In fact, from the point of view of cardiovascular medicine, it may be appropriate to say, “diabetes is a cardiovascular disease.

Rapid ocular responses are modulated by bottom-up-driven auditory salience

Despite the prevalent use of alerting sounds in alarms and human–machine interface systems and the long-hypothesized role of the auditory system as the brain's “early warning system,” we have only a rudimentary understanding of what determines auditory salience—the automatic attraction of attention by sound—and which brain mechanisms underlie this process. A major roadblock has been the lack of a robust, objective means of quantifying sound-driven attentional capture. Here we demonstrate that: (1) a reliable salience scale can be obtained from crowd-sourcing (N = 911), (2) acoustic roughness appears to be a driving feature behind this scaling, consistent with previous reports implicating roughness in the perceptual distinctiveness of sounds, and (3) crowd-sourced auditory salience correlates with objective autonomic measures. Specifically, we show that a salience ranking obtained from online raters correlated robustly with the superior colliculus-mediated ocular freezing response, microsaccadic inhibition (MSI), measured in naive, passively listening human participants (of either sex). More salient sounds evoked earlier and larger MSI, consistent with a faster orienting response. These results are consistent with the hypothesis that MSI reflects a general reorienting response that is evoked by potentially behaviorally important events regardless of their modality. SIGNIFICANCE STATEMENT Microsaccades are small, rapid, fixational eye movements that are measurable with sensitive eye-tracking equipment. We reveal a novel, robust link between microsaccade dynamics and the subjective salience of brief sounds (salience rankings obtained from a large number of participants in an online experiment): Within 300 ms of sound onset, the eyes of naive, passively listening participants demonstrate different microsaccade patterns as a function of the sound's crowd-sourced salience. These results position the superior colliculus (hypothesized to underlie microsaccade generation) as an important brain area to investigate in the context of a putative multimodal salience hub. They also demonstrate an objective means for quantifying auditory salience

Characterization of Karyopherin Cargoes Reveals Unique Mechanisms of Kap121p-Mediated Nuclear Import

In yeast there are at least 14 members of the β-karyopherin protein family that govern the movement of a diverse set of cargoes between the nucleus and cytoplasm. Knowledge of the cargoes carried by each karyopherin and insight into the mechanisms of transport are fundamental to understanding constitutive and regulated transport and elucidating how they impact normal cellular functions. Here, we have focused on the identification of nuclear import cargoes for the essential yeast β-karyopherin, Kap121p. Using an overlay blot assay and coimmunopurification studies, we have identified 30 putative Kap121p cargoes. Among these were Nop1p and Sof1p, two essential trans-acting protein factors required at the early stages of ribosome biogenesis. Characterization of the Kap121p-Nop1p and Kap121p-Sof1p interactions demonstrated that, in addition to lysine-rich nuclear localization signals (NLSs), Kap121p recognizes a unique class of signals distinguished by the abundance of arginine and glycine residues and consequently termed rg-NLSs. Kap104p is also known to recognize rg-NLSs, and here we show that it compensates for the loss of Kap121p function. Sof1p is also transported by Kap121p; however, its import can be mediated by a piggyback mechanism with Nop1p bridging the interaction between Sof1p and Kap121p. Together, our data elucidate additional levels of complexity in these nuclear transport pathways