Mutations in PI(3,5)P2 Signaling and Neurodegeneration in Mouse and Human.

Phosphoinositides are signaling lipids that are localized to the cytoplasmic surface of cellular membranes. Phosphoinositides regulate vesicle trafficking through unique tagging of membrane compartments and recruitment of effector proteins. PI(3,5)P2 is a minor phosphoinositide in yeast and mammalian cells. PI3P and PI(3,5)P2 function in the endosome/lysosome pathway. Early endosome membranes contain PI3P, while late endsomes/lysosomes contain PI(3,5)P2. I now report characterization of the pale tremor mouse, a spontaneous mutant characterized by neurodegeneration, diluted pigmentation, and lethality by six weeks of age. Genetic analysis identified a retroviral insertion that generated a null mutation in the gene Fig4, which encodes the 5-phosphatase that dephosphorylates PI(3,5)P2 to PI3P. The level of PI(3,5)P2 is reduced in cultured fibroblasts and there is accumulation of enlarged cytoplasmic vesicles containing the late endosome/lysosome marker LAMP2. The spontaneous mouse mutant ingls is also characterized by neurodegeneration, diluted pigmentation and juvenile lethality. I identified the causal mutation as the amino acid substitution L156R in Vac14, an activator of the kinase Fab1 that phosphorylates PI3P to generate PI(3,5)P2. ingls fibroblasts also exhibit reduced PI(3,5)P2 and accumulation of enlarged vesicles. The L156R mutation disrupts the interaction of Vac14 and Fab1, localizing the interaction site and demonstrating the importance of Fab1 activation in vivo. pale tremor mice exhibit peripheral neuropathy and early loss of DRG neurons. I screened 95 patients with Charcot-Marie-Tooth disease (CMT) by exon amplification and sequencing, and identified 4 patients carrying compound heterozygous pathogenic mutations in FIG4. Each patient carried one null allele and the missense mutation, I41T, which exhibits partial function in a yeast assay. To model this disorder, designated CMT4J, a transgenic mouse model carrying the I41T mutation was produced. CMT4J are healthy beyond 7 months of age, demonstrating that the I41T allele retains significant function in vivo. This thesis demonstrates the significant role of PI(3,5)P2 signaling in mammalian neurons, and expands the range of neurological disease related to phosphoinositide metabolism.PhDHuman GeneticsUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttp://deepblue.lib.umich.edu/bitstream/2027.42/61594/1/cychow_1.pd

Killing Vector Fields in Three Dimensions: A Method to Solve Massive Gravity Field Equations

Killing vector fields in three dimensions play important role in the construction of the related spacetime geometry. In this work we show that when a three dimensional geometry admits a Killing vector field then the Ricci tensor of the geometry is determined in terms of the Killing vector field and its scalars. In this way we can generate all products and covariant derivatives at any order of the ricci tensor. Using this property we give ways of solving the field equations of Topologically Massive Gravity (TMG) and New Massive Gravity (NMG) introduced recently. In particular when the scalars of the Killing vector field (timelike, spacelike and null cases) are constants then all three dimensional symmetric tensors of the geometry, the ricci and einstein tensors, their covariant derivatives at all orders, their products of all orders are completely determined by the Killing vector field and the metric. Hence the corresponding three dimensional metrics are strong candidates of solving all higher derivative gravitational field equations in three dimensions.Comment: 25 pages, some changes made and some references added, to be published in Classical and Quantum Gravit

Can a renal nurse assess fluid status using ultrasound on the inferior vena cava? A cross-sectional interrater study: Ultrasound on the inferior vena cava

Introduction: Ultrasound of the inferior vena cava (IVC-US) has been used to estimate intravascular volume status and fluid removal during a hemodialysis session. Usually, renal nurses rely on other, imprecise methods to determine ultrafiltration. To date, no study has examined whether renal nurses can reliably perform ultrasound for volume assessment and for potential prevention of intradialytic hypotension. This pilot study aimed to determine if a renal nurse could master the skill of performing and correctly interpreting Point of Care Ultrasound on patients receiving hemodialysis. Methods: After receiving theoretical training and performing 100 training scans, a renal nurse performed 60 ultrasound scans on 10 patients. These were categorized by the nurse into hypovolemic, euvolemic, or hypervolemic through measurement of the maximal diameter and degree of collapse of the IVC. Scans were subsequently assessed for adequacy and quality by two sonologists, who were blinded to each other\u27s and the nurse\u27s results. Findings: The interrater reliability of 60 scans was good, with intraclass correlation 0.79 (95% confidence interval (CI) =0.63–0.87) and with a good interrater agreement for the following estimation of intravascular volume (Cohen\u27s weighted Kappa κw = 0.62), when comparing the nurse to an expert sonographer. Discussion: A renal nurse can reliably perform ultrasound of the IVC in hemodialysis patients, obtaining high quality scans for volume assessment of hemodialysis patients. This novel approach could be more routinely applied by other renal nurses to obtain objective measures of patient volume status in the dialysis setting

All stationary axi-symmetric local solutions of topologically massive gravity

We classify all stationary axi-symmetric solutions of topologically massive gravity into Einstein, Schr\"odinger, warped and generic solutions. We construct explicitly all local solutions in the first three sectors and present an algorithm for the numerical construction of all local solutions in the generic sector. The only input for this algorithm is the value of one constant of motion if the solution has an analytic centre, and three constants of motion otherwise. We present several examples, including soliton solutions that asymptote to warped AdS.Comment: 42 pages, 9 figures. v2: Changed potentially confusing labelling of one sector, added references. v3: Minor changes, matches published versio

A Drosophila screen identifies NKCC1 as a modifier of NGLY1 deficiency

N-Glycanase 1 (NGLY1) is a cytoplasmic deglycosylating enzyme. Loss-of-function mutations in the NGLY1 gene cause NGLY1 deficiency, which is characterized by developmental delay, seizures, and a lack of sweat and tears. To model the phenotypic variability observed among patients, we crossed a Drosophila model of NGLY1 deficiency onto a panel of genetically diverse strains. The resulting progeny showed a phenotypic spectrum from 0 to 100% lethality. Association analysis on the lethality phenotype, as well as an evolutionary rate covariation analysis, generated lists of modifying genes, providing insight into NGLY1 function and disease. The top association hit was Ncc69 (human NKCC1/2), a conserved ion transporter. Analyses in NGLY1-/- mouse cells demonstrated that NKCC1 has an altered average molecular weight and reduced function. The misregulation of this ion transporter may explain the observed defects in secretory epithelium function in NGLY1 deficiency patients

Holographic Dual of Linear Dilaton Black Hole in Einstein-Maxwell-Dilaton-Axion Gravity

Motivated by the recently proposed Kerr/CFT correspondence, we investigate the holographic dual of the extremal and non-extremal rotating linear dilaton black hole in Einstein-Maxwell-Dilaton-Axion Gravity. For the case of extremal black hole, by imposing the appropriate boundary condition at spatial infinity of the near horizon extremal geometry, the Virasoro algebra of conserved charges associated with the asymptotic symmetry group is obtained. It is shown that the microscopic entropy of the dual conformal field given by Cardy formula exactly agrees with Bekenstein-Hawking entropy of extremal black hole. Then, by rewriting the wave equation of massless scalar field with sufficient low energy as the SL(2, R)$_L$$\times$SL(2, R)$_R$ Casimir operator, we find the hidden conformal symmetry of the non-extremal linear dilaton black hole, which implies that the non-extremal rotating linear dilaton black hole is holographically dual to a two dimensional conformal field theory with the non-zero left and right temperatures. Furthermore, it is shown that the entropy of non-extremal black hole can be reproduced by using Cardy formula.Comment: 15 pages, no figure, published versio

Pathogenic Mechanism of the FIG4 Mutation Responsible for Charcot-Marie-Tooth Disease CMT4J

CMT4J is a severe form of Charcot-Marie-Tooth neuropathy caused by mutation of the phosphoinositide phosphatase FIG4/SAC3. Affected individuals are compound heterozygotes carrying the missense allele FIG4-I41T in combination with a null allele. Analysis using the yeast two-hybrid system demonstrated that the I41T mutation impairs interaction of FIG4 with the scaffold protein VAC14. The critical role of this interaction was confirmed by the demonstration of loss of FIG4 protein in VAC14 null mice. We developed a mouse model of CMT4J by expressing a Fig4-I41T cDNA transgene on the Fig4 null background. Expression of the mutant transcript at a level 5× higher than endogenous Fig4 completely rescued lethality, whereas 2× expression gave only partial rescue, providing a model of the human disease. The level of FIG4-I41T protein in transgenic tissues is only 2% of that predicted by the transcript level, as a consequence of the protein instability caused by impaired interaction of the mutant protein with VAC14. Analysis of patient fibroblasts demonstrated a comparably low level of mutant I41T protein. The abundance of FIG4-I41T protein in cultured cells is increased by treatment with the proteasome inhibitor MG-132. The data demonstrate that FIG4-I41T is a hypomorphic allele encoding a protein that is unstable in vivo. Expression of FIG4-I41T protein at 10% of normal level is sufficient for long-term survival, suggesting that patients with CMT4J could be treated by increased production or stabilization of the mutant protein. The transgenic model will be useful for testing in vivo interventions to increase the abundance of the mutant protein

VAC14 nucleates a protein complex essential for the acute interconversion of PI3P and PI(3,5)P2 in yeast and mouse

The signalling lipid PI(3,5)P2 is generated on endosomes and regulates retrograde traffic to the trans-Golgi network. Physiological signals regulate rapid, transient changes in PI(3,5)P2 levels. Mutations that lower PI(3,5)P2 cause neurodegeneration in human patients and mice. The function of Vac14 in the regulation of PI(3,5)P2 was uncharacterized previously. Here, we predict that yeast and mammalian Vac14 are composed entirely of HEAT repeats and demonstrate that Vac14 exerts an effect as a scaffold for the PI(3,5)P2 regulatory complex by direct contact with the known regulators of PI(3,5)P2: Fig4, Fab1, Vac7 and Atg18. We also report that the mouse mutant ingls (infantile gliosis) results from a missense mutation in Vac14 that prevents the association of Vac14 with Fab1, generating a partial complex. Analysis of ingls and two additional mutants provides insight into the organization of the PI(3,5)P2 regulatory complex and indicates that Vac14 mediates three distinct mechanisms for the rapid interconversion of PI3P and PI(3,5)P2. Moreover, these studies show that the association of Fab1 with the complex is essential for viability in the mouse

A Unified Approach to Variational Derivatives of Modified Gravitational Actions

Our main aim in this paper is to promote the coframe variational method as a unified approach to derive field equations for any given gravitational action containing the algebraic functions of the scalars constructed from the Riemann curvature tensor and its contractions. We are able to derive a master equation which expresses the variational derivatives of the generalized gravitational actions in terms of the variational derivatives of its constituent curvature scalars. Using the Lagrange multiplier method relative to an orthonormal coframe, we investigate the variational procedures for modified gravitational Lagrangian densities in spacetime dimensions $n\geqslant 3$. We study well-known gravitational actions such as those involving the Gauss-Bonnet and Ricci-squared, Kretchmann scalar, Weyl-squared terms and their algebraic generalizations similar to generic $f(R)$ theories and the algebraic generalization of sixth order gravitational Lagrangians. We put forth a new model involving the gravitational Chern-Simons term and also give three dimensional New massive gravity equations in a new form in terms of the Cotton 2-form

Measurement of the cross-section and charge asymmetry of WW bosons produced in proton-proton collisions at s=8\sqrt{s}=8 TeV with the ATLAS detector

This paper presents measurements of the $W^+ \rightarrow \mu^+\nu$ and $W^- \rightarrow \mu^-\nu$ cross-sections and the associated charge asymmetry as a function of the absolute pseudorapidity of the decay muon. The data were collected in proton--proton collisions at a centre-of-mass energy of 8 TeV with the ATLAS experiment at the LHC and correspond to a total integrated luminosity of 20.2~\mbox{fb^{-1}}. The precision of the cross-section measurements varies between 0.8% to 1.5% as a function of the pseudorapidity, excluding the 1.9% uncertainty on the integrated luminosity. The charge asymmetry is measured with an uncertainty between 0.002 and 0.003. The results are compared with predictions based on next-to-next-to-leading-order calculations with various parton distribution functions and have the sensitivity to discriminate between them.Comment: 38 pages in total, author list starting page 22, 5 figures, 4 tables, submitted to EPJC. All figures including auxiliary figures are available at https://atlas.web.cern.ch/Atlas/GROUPS/PHYSICS/PAPERS/STDM-2017-13