Infectious SIV resides in adipose tissue and induces metabolic defects in chronically infected rhesus macaques

Additional file 1. General method for isolation of stromal-vascular-fraction (AT-SVF) cells from adipose tissue of rhesus macaques, and subsequent analyses. (A) 30-60 mins collagenase digestion of solid adipose tissue samples from rhesus macaques is followed by washing and centrifugation, allowing for separation of mature adipocytes (floater fraction) from the stromal-vascular-fraction (AT-SVF) cells. AT-SVF cells were then analyzed by flow cytometry, nested PCR, and viral outgrowth assays, and floater fraction adipocytes analyzed for mRNA expression. (B) Sample flow cytometry gating schemes for examination of AT-SVF T cells, NKT cells, macrophages, and B cells

Hemodynamic Effects of Anthrax Toxins in the Rabbit Model and the Cardiac Pathology Induced by Lethal Toxin

Anthrax lethal toxin (LeTx) and edema toxin (EdTx) have been shown to alter hemodynamics in the rodent model, while LeTx primarily is reported to induce extensive tissue pathology. However, the rodent model has limitations when used for comparison to higher organisms such as humans. The rabbit model, on the other hand, has gained recognition as a useful model for studying anthrax infection and its pathophysiological effects. In this study, we assessed the hemodynamic effects of lethal toxin (LeTx) and edema toxin (EdTx) in the rabbit model using physiologically relevant amounts of the toxins. Moreover, we further examine the pathological effects of LeTx on cardiac tissue. We intravenously injected Dutch-belted rabbits with either low-dose and high-dose recombinant LeTx or a single dose of EdTx. The animals’ heart rate and mean arterial pressure were continuously monitored via telemetry until either 48 or 72 h post-challenge. Additional animals challenged with LeTx were used for cardiac troponin I (cTnI) quantitation, cardiac histopathology, and echocardiography. LeTx depressed heart rate at the lower dose and mean arterial pressure (MAP) at the higher dose. EdTx, on the other hand, temporarily intensified heart rate while lowering MAP. Both doses of LeTx caused cardiac pathology with the higher dose having a more profound effect. Lastly, left-ventricular dilation due to LeTx was not apparent at the given time-points. Our study demonstrates the hemodynamic effects of anthrax toxins, as well as the pathological effects of LeTx on the heart in the rabbit model, and it provides further evidence for the toxins’ direct impact on the heart

Phospholipase A2-activating protein is associated with a novel form of leukoencephalopathy

Leukoencephalopathies are a group of white matter disorders related to abnormal formation, maintenance, and turnover of myelin in the central nervous system. These disorders of the brain are categorized according to neuroradiological and pathophysiological criteria. Herein, we have identified a unique form of leukoencephalopathy in seven patients presenting at ages 2 to 4 months with progressive microcephaly, spastic quadriparesis, and global developmental delay. Clinical, metabolic, and imaging characterization of seven patients followed by homozygosity mapping and linkage analysis were performed. Next generation sequencing, bioinformatics, and segregation analyses followed, to determine a loss of function sequence variation in the phospholipase A2-activating protein encoding gene (PLAA). Expression and functional studies of the encoded protein were performed and included measurement of prostaglandin E2 and cytosolic phospholipase A2 activity in membrane fractions of fibroblasts derived from patients and healthy controls. Plaa-null mice were generated and prostaglandin E2 levels were measured in different tissues. The novel phenotype of our patients segregated with a homozygous loss-of-function sequence variant, causing the substitution of leucine at position 752 to phenylalanine, in PLAA, which causes disruption of the protein's ability to induce prostaglandin E2 and cytosolic phospholipase A2 synthesis in patients' fibroblasts. Plaa-null mice were perinatal lethal with reduced brain levels of prostaglandin E2 The non-functional phospholipase A2-activating protein and the associated neurological phenotype, reported herein for the first time, join other complex phospholipid defects that cause leukoencephalopathies in humans, emphasizing the importance of this axis in white matter development and maintenance

SPIRAL II Project (electron option) - Preliminary Design Study

This document presents a Preliminary Design Study (PDS) of the electron option of the SPIRAL II project

Performance of an endcap prototype of the Atlas accordion electromagnetic calorimeter

The design and construction of a lead-liquid argon endcap calorimeter prototype using an accordion geometry and conceived as a sector of the inner wheel of the endcap calorimeter of the future ATLAS experiment at the LHC is described. The performance obtained using electron beam data is presented. The main results are an energy resolution with a sampling term below $11\%/\sqrt{E(\rm GeV)}$ and a small local constant term, a good linearity of the response with the incident energy and a global constant term of 0.8\% over an extended area in the rapidity range of $2.2\eta 2.9$. These properties make the design suitable for the ATLAS electromagnetic endcap calorimeter

Construction and test of a fine-grained liquid argon preshower prototype

A separate liquid argon preshower detector consisting of two layers featuring a fine granularity of 2.5~10$^{\mathrm{-3}}$ was studied by the RD3 collaboration. A prototype covering approximately 0.8 in pseudo-rapidity and 9 degrees in azimuth was built and tested at CERN in July 94. CMOS and GaAs VLSI preamplifiers were designed and tested for this occasion. The combined response of this detector and an accordion electromagnetic calorimeter prototype to muons, electrons and photons is presented. For minimum ionizing tracks a signal-to-noise ratio of 4.5 per preshower layer was measured. Above 150~GeV the space resolution for electrons is better than 250~$\mu$m in both directions. The precision on the electromagnetic shower direction, determined together with the calorimeter, is better than 4 mrad above 50~GeV. It is concluded that the preshower detector would adequately fulfil its role for future operation at CERN Large Hadron Collider

Therapeutic targeting of membrane-associated GRP78 in leukemia and lymphoma : preclinical efficacy in vitro and formal toxicity study of BMTP-78 in rodents and primates

Translation of drug candidates into clinical settings requires demonstration of preclinical efficacy and formal toxicology analysis for filling an Investigational New Drug (IND) application with the US Food and Drug Administration (FDA). Here, we investigate the membrane-associated glucose response protein 78 (GRP78) as a therapeutic target in leukemia and lymphoma. We evaluated the efficacy of the GRP78-targeted proapoptotic drug bone metastasis targeting peptidomimetic 78 (BMTP-78), a member of the D (KLAKLAK)2-containing class of agents. BMTP-78 was validated in cells from patients with acute myeloid leukemia and in a panel of human leukemia and lymphoma cell lines, where it induced dose-dependent cytotoxicity in all samples tested. Based on the in vitro efficacy of BMTP-78, we performed formal good laboratory practice toxicology studies in both rodents (mice and rats) and nonhuman primates (cynomolgus and rhesus monkeys). These analyses represent required steps towards an IND application of BMTP-78 for theranostic first-in-human clinical trials.Peer reviewe

Performance of a large scale prototype of the ATLAS accordion electromagnetic calorimeter

A 2 m long prototype of a lead-liquid argon electromagnetic calorimeter with accordion-shaped electrodes, conceived as a sector of the barrel calorimeter of the future ATLAS experiment at the LHC, has been tested with electron and pion beams in the energy range 10 to 287 GeV. A sampling term of 10%/root E(GeV) was obtained for electrons in the rapidity range 0 < eta < 1, while the constant term measured over an area of about 1 m(2) is 0.69%. With a cell size of 2.7 cm the position resolution is. about 4 mm/root E(GeV)