Ligand Mediated Sequestering of Integrins in Raft-Mimicking Lipid Mixtures: The Role of Bilayer Asymmetry and Cholesterol Content

poster abstractLipid microdomains play an important functional role in plasma membranes. However, the small size and transient nature of lipid/membrane heterogeneities in the plasma membrane make characterization of microdomains and microdomain-related membrane processes quite challenging. To address this issue, we recently introduced a powerful model membrane system that allows the investigation of membrane protein sequestering and oligomerization in raft-mimicking lipid mixtures using combined confocal fluorescence spectroscopy, photon counting histogram (PCH), and epifluorescence microscopy. Our experiments on bilayer-spanning domains showed that αvβ3 and α5β1 integrins predominantly exist as monomers and sequester preferentially to the liquid-disordered (ld) phase in the absence of ligands. Notably, addition of vitronectin (αvβ3) and fibronectin (α5β1) caused substantial translocations of integrins into the liquid-ordered (lo) phase without altering receptor oligomerization state. Here we expand our previous studies and report on the sequestering and oligomerization state of αvβ3 and α5β1 in asymmetric bilayer compositions containing coexisting lo and ld phases located exclusively in the top leaflet of the bilayer (bottom leaflet shows only ld phase). Remarkably, in such a membrane environment, both integrins show a higher affinity for the top leaflet-restricted lo domains in the absence of their respective ligands. A slight change in the integrin sequestration was observed after addition of their respective ligands. We also present experimental findings, which show that cholesterol content has a substantial influence on integrin sequestering and oligomerization in raft-mimicking lipid mixtures. The described experimental results highlight the potential importance of membrane asymmetry and lipid composition in the sequestering of membrane proteins in biological membranes

Augmentation of abscisic acid (ABA) levels by drought does not induce short-term stomatal sensitivity to CO2 in two divergent conifer species

The stomata of conifers display very little short-term response to changes in atmospheric CO2 concentration (Ca), whereas the stomatal responses of angiosperms to Ca increase in response to water stress. This behaviour of angiosperm stomata appears to be dependent on foliar levels of abscisic acid (ABAf). Here two alternative explanations for the stomatal insensitivity of conifers to Ca are tested: that conifers have either low ABAf or a higher or absent threshold for ABA-induced sensitivity. The responsiveness of stomatal conductance (gs) to a sequence of transitions in Ca (386, 100, and 600 μmol mol−1) was recorded over a range of ABAf in an angiosperm and two divergent conifer species. The different ABA levels were induced by a mild drought cycle. Although the angiosperm and conifer species showed similar proportional increases in ABAf following drought, conifer stomata remained insensitive to changes in Ca whereas angiosperm stomata showed enhanced sensitivity with increasing ABAf. The conifers, however, had much higher ABAf prior to drought than the angiosperm species, suggesting that non-sensitivity to Ca in these conifers was due to an absent or inactive response/signalling pathway rather than insufficient ABAf

The Real Symplectic Groups in Quantum Mechanics and Optics

text of abstract (We present a utilitarian review of the family of matrix groups $Sp(2n,\Re)$, in a form suited to various applications both in optics and quantum mechanics. We contrast these groups and their geometry with the much more familiar Euclidean and unitary geometries. Both the properties of finite group elements and of the Lie algebra are studied, and special attention is paid to the so-called unitary metaplectic representation of $Sp(2n,\Re)$. Global decomposition theorems, interesting subgroups and their generators are described. Turning to $n$-mode quantum systems, we define and study their variance matrices in general states, the implications of the Heisenberg uncertainty principles, and develop a U(n)-invariant squeezing criterion. The particular properties of Wigner distributions and Gaussian pure state wavefunctions under $Sp(2n,\Re)$ action are delineated.)Comment: Review article 43 pages, revtex, no figures, replaced because somefonts were giving problem in autometic ps generatio

Power Law versus Exponential State Transition Dynamics: Application to Sleep-Wake Architecture

BACKGROUND: Despite the common experience that interrupted sleep has a negative impact on waking function, the features of human sleep-wake architecture that best distinguish sleep continuity versus fragmentation remain elusive. In this regard, there is growing interest in characterizing sleep architecture using models of the temporal dynamics of sleep-wake stage transitions. In humans and other mammals, the state transitions defining sleep and wake bout durations have been described with exponential and power law models, respectively. However, sleep-wake stage distributions are often complex, and distinguishing between exponential and power law processes is not always straightforward. Although mono-exponential distributions are distinct from power law distributions, multi-exponential distributions may in fact resemble power laws by appearing linear on a log-log plot. METHODOLOGY/PRINCIPAL FINDINGS: To characterize the parameters that may allow these distributions to mimic one another, we systematically fitted multi-exponential-generated distributions with a power law model, and power law-generated distributions with multi-exponential models. We used the Kolmogorov-Smirnov method to investigate goodness of fit for the "incorrect" model over a range of parameters. The "zone of mimicry" of parameters that increased the risk of mistakenly accepting power law fitting resembled empiric time constants obtained in human sleep and wake bout distributions. CONCLUSIONS/SIGNIFICANCE: Recognizing this uncertainty in model distinction impacts interpretation of transition dynamics (self-organizing versus probabilistic), and the generation of predictive models for clinical classification of normal and pathological sleep architecture

Thinking the unthinkable: Imagining an ‘un-American,’ Girl-friendly, Women- and Trans-Inclusive Alternative for Baseball

The purpose of this article is twofold: to capture the injustice inherent in the gendered bifurcation of baseball and softball via the prism of critical feminist sport studies; and to begin to imagine a girl-friendly/women-and trans-inclusive future for baseball that is less fertile for cooptation into post-911 United States security state discourses. In this article I link the "unthinkability" of the occupational segregation of baseball in North America to the dominance of the ideology of the two sex system and European disasporic morality. To illustrate the extent of this occupational segregation via the gendered bifurcation of baseball and softball, I draw on feminist sport studies to examine the exemplars or "texts" of three Canadian brother/sister baseball softball duos: Jason Bay and Lauren Bay Regula; Brett and Danielle Lawrie; and Mathew and Katie Reyes

Proteolytic cleavage and loss of function of biologic agents that neutralize tumor necrosis factor in the mucosa of patients with inflammatory bowel disease

BACKGROUND & AIMS: Many patients with inflammatory bowel disease (IBD) fail to respond to anti–tumor necrosis factor (TNF) agents such as infliximab and adalimumab, and etanercept is not effective for treatment of Crohn’s disease. Activated matrix metalloproteinase 3 (MMP3) and MMP12, which are increased in inflamed mucosa of patients with IBD, have a wide range of substrates, including IgG1. TNFneutralizing agents act in inflamed tissues; we investigated the effects of MMP3, MMP12, and mucosal proteins from IBD patients on these drugs. METHODS: Biopsy specimens from inflamed colon of 8 patients with Crohn’s disease and 8 patients with ulcerative colitis, and from normal colon of 8 healthy individuals (controls), were analyzed histologically, or homogenized and proteins were extracted. We also analyzed sera from 29 patients with active Crohn’s disease and 33 patients with active ulcerative colitis who were candidates to receive infliximab treatment. Infliximab, adalimumab, and etanercept were incubated with mucosal homogenates from patients with IBD or activated recombinant human MMP3 or MMP12 and analyzed on immunoblots or in luciferase reporter assays designed to measure TNF activity. IgG cleaved by MMP3 or MMP12 and antihinge autoantibodies against neo-epitopes on cleaved IgG were measured in sera from IBD patients who subsequently responded (clinical remission and complete mucosal healing) or did not respond to infliximab. RESULTS: MMP3 and MMP12 cleaved infliximab, adalimumab, and etanercept, releasing a 32-kilodalton Fc monomer. After MMP degradation, infliximab and adalimumab functioned as F(ab’)2 fragments, whereas cleaved etanercept lost its ability to neutralize TNF. Proteins from the mucosa of patients with IBD reduced the integrity and function of infliximab, adalimumab, and etanercept. TNF-neutralizing function was restored after incubation of the drugs with MMP inhibitors. Serum levels of endogenous IgG cleaved by MMP3 and MMP12, and antihinge autoantibodies against neo-epitopes of cleaved IgG, were higher in patients who did not respond to treatment vs responders. CONCLUSIONS: Proteolytic degradation may contribute to the nonresponsiveness of patients with IBD to anti-TNF agents