Amino-terminal cysteine residues of RGS16 are required for palmitoylation and modulation of G(i)- and G(q)-mediated signaling

RGS proteins (Regulators of G protein Signaling) are a recently discovered family of proteins that accelerate the GTPase activity of heterotrimeric G protein α subunits of the i, q, and 12 classes. The proteins share a homologous core domain but have divergent amino-terminal sequences that are the site of palmitoylation for RGS-GAIP and RGS4. We investigated the function of palmitoylation for RGS16, which shares conserved amino-terminal cysteines with RGS4 and RGS5. Mutation of cysteine residues at residues 2 and 12 blocked the incorporation of [3H]palmitate into RGS16 in metabolic labeling studies of transfected cells or into purified RGS proteins in a cell-free palmitoylation assay. The purified RGS16 proteins with the cysteine mutations were still able to act as GTPase-activating protein for Giα. Inhibition or a decrease in palmitoylation did not significantly change the amount of protein that was membrane-associated. However, palmitoylation-defective RGS16 mutants demonstrated impaired ability to inhibit both Gi- and Gq-linked signaling pathways when expressed in HEK293T cells. These findings suggest that the amino-terminal region of RGS16 may affect the affinity of these proteins for Gα subunits in vivo or that palmitoylation localizes the RGS protein in close proximity to Gα subunits on cellular membranes

A Simple Passive Scalar Advection-Diffusion Model

This paper presents a simple, one-dimensional model of a randomly advected passive scalar. The model exhibits anomalous inertial range scaling for the structure functions constructed from scalar differences. The model provides a simple computational test for recent ideas regarding closure and scaling for randomly advected passive scalars. Results suggest that high order structure function scaling depends on the largest velocity eddy size, and hence scaling exponents may be geometry-dependent and non-universal.Comment: 30 pages, 11 figure

Passive Scalar: Scaling Exponents and Realizability

An isotropic passive scalar field $T$ advected by a rapidly-varying velocity field is studied. The tail of the probability distribution $P(\theta,r)$ for the difference $\theta$ in $T$ across an inertial-range distance $r$ is found to be Gaussian. Scaling exponents of moments of $\theta$ increase as $\sqrt{n}$ or faster at large order $n$, if a mean dissipation conditioned on $\theta$ is a nondecreasing function of $|\theta|$. The $P(\theta,r)$ computed numerically under the so-called linear ansatz is found to be realizable. Some classes of gentle modifications of the linear ansatz are not realizable.Comment: Substantially revised to conform with published version. Revtex (4 pages) with 2 postscript figures. Send email to [email protected]

Cas9+ conditionally-immortalized macrophages as a tool for bacterial pathogenesis and beyond.

Macrophages play critical roles in immunity, development, tissue repair, and cancer, but studies of their function have been hampered by poorly-differentiated tumor cell lines and genetically-intractable primary cells. Here we report a facile system for genome editing in non-transformed macrophages by differentiating ER-Hoxb8 myeloid progenitors from Cas9-expressing transgenic mice. These conditionally immortalized macrophages (CIMs) retain characteristics of primary macrophages derived from the bone marrow yet allow for easy genetic manipulation and a virtually unlimited supply of cells. We demonstrate the utility of this system for dissection of host genetics during intracellular bacterial infection using two important human pathogens: Listeria monocytogenes and Mycobacterium tuberculosis

Self-organising management of Grid environments

This paper presents basic concepts, architectural principles and algorithms for efficient resource and security management in cluster computing environments and the Grid. The work presented in this paper is funded by BTExacT and the EPSRC project SO-GRM (GR/S21939)

Wave Propagation in Gravitational Systems: Completeness of Quasinormal Modes

The dynamics of relativistic stars and black holes are often studied in terms of the quasinormal modes (QNM's) of the Klein-Gordon (KG) equation with different effective potentials $V(x)$. In this paper we present a systematic study of the relation between the structure of the QNM's of the KG equation and the form of $V(x)$. In particular, we determine the requirements on $V(x)$ in order for the QNM's to form complete sets, and discuss in what sense they form complete sets. Among other implications, this study opens up the possibility of using QNM expansions to analyse the behavior of waves in relativistic systems, even for systems whose QNM's do {\it not} form a complete set. For such systems, we show that a complete set of QNM's can often be obtained by introducing an infinitesimal change in the effective potential

Using diffusion tensor imaging to identify corticospinal tract projection patterns in children with unilateral spastic cerebral palsy.

AIM: To determine whether diffusion tensor imaging (DTI) can be an independent assessment for identifying the corticospinal tract (CST) projecting from the more-affected motor cortex in children with unilateral spastic cerebral palsy (CP). METHOD: Twenty children with unilateral spastic CP participated in this study (16 males, four females; mean age 9y 2mo [standard deviation (SD) 3y 2mo], Manual Ability Classification System [MACS] level I-III). We used DTI tractography to reconstruct the CST projecting from the more-affected motor cortex. We mapped the motor representation of the more-affected hand by stimulating the more- and the less-affected motor cortex measured with single-pulse transcranial magnetic stimulation (TMS). We then verified the presence or absence of the contralateral CST by comparing the TMS map and DTI tractography. Fisher's exact test was used to determine the association between findings of TMS and DTI. RESULTS: DTI tractography successfully identified the CST controlling the more-affected hand (sensitivity=82%, specificity=78%). INTERPRETATION: Contralateral CST projecting from the lesioned motor cortex assessed by DTI is consistent with findings of TMS mapping. Since CST connectivity may be predictive of response to certain upper extremity treatments, DTI-identified CST connectivity may potentially be valuable for determining such connectivity where TMS is unavailable or inadvisable for children with seizures.K08 NS073796 - NINDS NIH HHS; TL1 RR024158 - NCRR NIH HHS; K01 NS062116 - NINDS NIH HHS; UL1 RR024156 - NCRR NIH HHS; KL2 RR024157 - NCRR NIH HHS; R01 HD076436 - NICHD NIH HHSPublished versio

High-Order Contamination in the Tail of Gravitational Collapse

It is well known that the late-time behaviour of gravitational collapse is {\it dominated} by an inverse power-law decaying tail. We calculate {\it higher-order corrections} to this power-law behaviour in a spherically symmetric gravitational collapse. The dominant ``contamination'' is shown to die off at late times as $M^2t^{-4}\ln(t/M)$. This decay rate is much {\it slower} than has been considered so far. It implies, for instance, that an `exact' (numerical) determination of the power index to within $\sim 1$ requires extremely long integration times of order $10^4 M$. We show that the leading order fingerprint of the black-hole electric {\it charge} is of order $Q^2t^{-4}$.Comment: 12 pages, 2 figure

Mode-coupling in rotating gravitational collapse: Gravitational and electromagnetic perturbations

We consider the late-time evolution of {\it gravitational} and electromagnetic perturbations in realistic {\it rotating} Kerr spacetimes. We give a detailed analysis of the mode-coupling phenomena in rotating gravitational collapse. A consequence of this phenomena is that the late-time tail is dominated by modes which, in general, may have an angular distribution different from the original one. In addition, we show that different types of fields have {\it different} decaying rates. This result turns over the traditional belief (which has been widely accepted during the last three decades) that the late-time tail of gravitational collapse is universal.Comment: 16 page