Heterologous prime-boost-boost immunisation of Chinese cynomolgus macaques using DNA and recombinant poxvirus vectors expressing HIV-1 virus-like particles

Background: There is renewed interest in the development of poxvirus vector-based HIV vaccines due to the protective effect observed with repeated recombinant canarypox priming with gp120 boosting in the recent Thai placebo-controlled trial. This study sought to investigate whether a heterologous prime-boost-boost vaccine regimen in Chinese cynomolgus macaques with a DNA vaccine and recombinant poxviral vectors expressing HIV virus-like particles bearing envelopes derived from the most prevalent clades circulating in sub-Saharan Africa, focused the antibody response to shared neutralising epitopes. Methods: Three Chinese cynomolgus macaques were immunised via intramuscular injections using a regimen composed of a prime with two DNA vaccines expressing clade A Env/clade B Gag followed by boosting with recombinant fowlpox virus expressing HIV-1 clade D Gag, Env and cholera toxin B subunit followed by the final boost with recombinant modified vaccinia virus Ankara expressing HIV-1 clade C Env, Gag and human complement protein C3d. We measured the macaque serum antibody responses by ELISA, enumerated T cell responses by IFN-gamma ELISpot and assessed seroneutralisation of HIV-1 using the TZM-bl beta-galactosidase assay with primary isolates of HIV-1. Results: This study shows that large and complex synthetic DNA sequences can be successfully cloned in a single step into two poxvirus vectors: MVA and FPV and the recombinant poxviruses could be grown to high titres. The vaccine candidates showed appropriate expression of recombinant proteins with the formation of authentic HIV virus-like particles seen on transmission electron microscopy. In addition the b12 epitope was shown to be held in common by the vaccine candidates using confocal immunofluorescent microscopy. The vaccine candidates were safely administered to Chinese cynomolgus macaques which elicited modest T cell responses at the end of the study but only one out of the three macaques elicited an HIV-specific antibody response. However, the antibodies did not neutralise primary isolates of HIV-1 or the V3-sensitive isolate SF162 using the TZM-bl b-galactosidase assay. Conclusions: MVA and FP9 are ideal replication-deficient viral vectors for HIV-1 vaccines due to their excellent safety profile for use in humans. This study shows this novel prime-boost-boost regimen was poorly immunogenic in Chinese cynomolgus macaques

Evaluation of HFC 245ca and HFC 236ea as foam blowing agents

Hydrochlorofluorocarbon (HCFC) 141b has been selected as the interim blowing agent for use in urethane insulations on NASA's Space Shuttle External Tank. Due to the expected limited commercial lifetime of this material, research efforts at the NASA Thermal Protection Systems Materials Research Laboratory at the Marshall Space Flight Center are now being devoted to the identification and development of alternatives with zero ozone depletion potential. Physical blowing agents identified to date have included hydrocarbons, fluorocarbons, hydrofluoroethers, and more predominantly, hydrofluorocarbons (HFCs). The majority of the HFC evaluations in industry have focused on the more readily available, low boiling candidates such as HFC 134a. Higher boiling HFC candidates that could be handled at ambient conditions and use current processing equipment would be more desirable. This paper will describe results from a research program of two such candidate HFC's performed as a cooperative effort between Martin Marietta Manned Space Systems, the U.S. Environmental Protection Agency, and Oak Ridge National Laboratories. The purpose of this effort was to perform a cursory evaluation of the developmental HFC's 245ca and 236ea as blowing agents in urethane based insulations. These two materials were selected from screening tests of 37 C2, C3, and C4 isomers based on physical properties, atmospheric lifetime, flammability, estimated toxicity, difficulty of synthesis, suitability for dual use as a refrigerant, and other factors. Solubility of the two materials in typical foam components was tested, pour foaming trials were performed, and preliminary data were gathered regarding foam insulation performance

Light hadron spectrum---MILC results with the Kogut-Susskind and Wilson actions

We present the current status of our ongoing calculations of the light hadron spectrum with both Kogut-Susskind (KS) and Wilson quarks in the valence or quenched approximation. We discuss KS quarks first and find that the chiral extrapolation is potentially the biggest source of systematic error. For the Wilson case, we focus on finite volume and source size effects at 6/g^2=5.7. We find no evidence to support the claim that there is a finite volume effect between N_s=16 and 24 of approximately 5%.Comment: 11 pages, 11 figures, LaTeX, uses espcrs2, epsf, Invited talk presented by S. Gottlieb at Lattice QCD on Parallel Computers, University of Tsukuba, March, 1997, to appear in the proceeding

Opportunities for lattice QCD in quark and lepton flavor physics

This document is one of a series of whitepapers from the USQCD collaboration. Here, we discuss opportunities for lattice QCD in quark and lepton flavor physics. New data generated at Belle II, LHCb, BES III, NA62, KOTO, and Fermilab E989, combined with precise calculations of the relevant hadronic physics, may reveal what lies beyond the Standard Model. We outline a path toward improvements of the precision of existing lattice-QCD calculations and discuss groundbreaking new methods that allow lattice QCD to access new observables.Comment: USQCD whitepape

3 dimensional modelling of early human brain development using optical projection tomography

BACKGROUND: As development proceeds the human embryo attains an ever more complex three dimensional (3D) structure. Analyzing the gene expression patterns that underlie these changes and interpreting their significance depends on identifying the anatomical structures to which they map and following these patterns in developing 3D structures over time. The difficulty of this task greatly increases as more gene expression patterns are added, particularly in organs with complex 3D structures such as the brain. Optical Projection Tomography (OPT) is a new technology which has been developed for rapidly generating digital 3D models of intact specimens. We have assessed the resolution of unstained neuronal structures within a Carnegie Stage (CS)17 OPT model and tested its use as a framework onto which anatomical structures can be defined and gene expression data mapped. RESULTS: Resolution of the OPT models was assessed by comparison of digital sections with physical sections stained, either with haematoxylin and eosin (H&E) or by immunocytochemistry for GAP43 or PAX6, to identify specific anatomical features. Despite the 3D models being of unstained tissue, peripheral nervous system structures from the trigeminal ganglion (~300 μm by ~150 μm) to the rootlets of cranial nerve XII (~20 μm in diameter) were clearly identifiable, as were structures in the developing neural tube such as the zona limitans intrathalamica (core is ~30 μm thick). Fourteen anatomical domains have been identified and visualised within the CS17 model. Two 3D gene expression domains, known to be defined by Pax6 expression in the mouse, were clearly visible when PAX6 data from 2D sections were mapped to the CS17 model. The feasibility of applying the OPT technology to all stages from CS12 to CS23, which encompasses the major period of organogenesis for the human developing central nervous system, was successfully demonstrated. CONCLUSION: In the CS17 model considerable detail is visible within the developing nervous system at a minimum resolution of ~20 μm and 3D anatomical and gene expression domains can be defined and visualised successfully. The OPT models and accompanying technologies for manipulating them provide a powerful approach to visualising and analysing gene expression and morphology during early human brain development

The QCD spectrum with three quark flavors

We present results from a lattice hadron spectrum calculation using three flavors of dynamical quarks - two light and one strange, and quenched simulations for comparison. These simulations were done using a one-loop Symanzik improved gauge action and an improved Kogut-Susskind quark action. The lattice spacings, and hence also the physical volumes, were tuned to be the same in all the runs to better expose differences due to flavor number. Lattice spacings were tuned using the static quark potential, so as a byproduct we obtain updated results for the effect of sea quarks on the static quark potential. We find indications that the full QCD meson spectrum is in better agreement with experiment than the quenched spectrum. For the 0++ (a0) meson we see a coupling to two pseudoscalar mesons, or a meson decay on the lattice.Comment: 38 pages, 20 figures, uses epsf. 5/29/01 revision responds to referee's Comments, changes pion fits and tables, and corrects Fig. 10 and some minor error