64 research outputs found

    Gp41-targeted antibodies restore infectivity of a fusion-deficient HIV-1 envelope glycoprotein

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    The HIV-1 envelope glycoprotein (Env) mediates viral entry via conformational changes associated with binding the cell surface receptor (CD4) and coreceptor (CCR5/CXCR4), resulting in subsequent fusion of the viral and cellular membranes. While the gp120 Env surface subunit has been extensively studied for its role in viral entry and evasion of the host immune response, the gp41 transmembrane glycoprotein and its role in natural infection are less well characterized. Here, we identified a primary HIV-1 Env variant that consistently supports \u3e300% increased viral infectivity in the presence of autologous or heterologous HIV-positive plasma. However, in the absence of HIV-positive plasma, viruses with this Env exhibited reduced infectivity that was not due to decreased CD4 binding. Using Env chimeras and sequence analysis, we mapped this phenotype to a change Q563R, in the gp41 heptad repeat 1 (HR1) region. We demonstrate that Q563R reduces viral infection by disrupting formation of the gp41 six-helix bundle required for virus-cell membrane fusion. Intriguingly, antibodies that bind cluster I epitopes on gp41 overcome this inhibitory effect, restoring infectivity to wild-type levels. We further demonstrate that the Q563R change increases HIV-1 sensitivity to broadly neutralizing antibodies (bNAbs) targeting the gp41 membrane-proximal external region (MPER). In summary, we identify an HIV-1 Env variant with impaired infectivity whose Env functionality is restored through the binding of host antibodies. These data contribute to our understanding of gp41 residues involved in membrane fusion and identify a mechanism by which host factors can alleviate a viral defect

    Impact of Sleep and Circadian Disruption on Energy Balance and Diabetes: A Summary of Workshop Discussions

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    A workshop was held at the National Institute for Diabetes and Digestive and Kidney Diseases with a focus on the impact of sleep and circadian disruption on energy balance and diabetes. The workshop identified a number of key principles for research in this area and a number of specific opportunities. Studies in this area would be facilitated by active collaboration between investigators in sleep/circadian research and investigators in metabolism/diabetes. There is a need to translate the elegant findings from basic research into improving the metabolic health of the American public. There is also a need for investigators studying the impact of sleep/circadian disruption in humans to move beyond measurements of insulin and glucose and conduct more in-depth phenotyping. There is also a need for the assessments of sleep and circadian rhythms as well as assessments for sleep-disordered breathing to be incorporated into all ongoing cohort studies related to diabetes risk. Studies in humans need to complement the elegant short-term laboratory-based human studies of simulated short sleep and shift work etc. with studies in subjects in the general population with these disorders. It is conceivable that chronic adaptations occur, and if so, the mechanisms by which they occur needs to be identified and understood. Particular areas of opportunity that are ready for translation are studies to address whether CPAP treatment of patients with pre-diabetes and obstructive sleep apnea (OSA) prevents or delays the onset of diabetes and whether temporal restricted feeding has the same impact on obesity rates in humans as it does in mice

    Flight Operations of Two Rapidly Assembled CubeSats with Commercial Infrared Cameras: The Rogue-Alpha,Beta Program

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    The Aerospace Corporation’s Rogue-alpha, betaprogram, co-funded by the Space and Missile Systems Center’s Development Corps, is a rapid prototyping effort that built and launched two 3-Unit CubeSats equipped with modified commercial IR camera payloads, laser communications and precision pointing capabilities in 18-months. Launched on 2 November 2019, the two spacecraft were released from the ISS Cygnus NG-12 robotic resupply spacecraft on 31 January 2020 into a circular 460-km, 52° inclined orbit. The two Rogue spacecraft are serving as testbeds for studying wide-field-of-view fast-framing imaging, on-orbit stellar calibration techniques for small IR payloads, and associated spacecraft flight operations. Precision pointing is enabled by three star sensors. High data rate sensor observations are enabled by the ultra-compact 200 Mbps lasercom system, which downlinks gigabytes of stored data during a single laser contact, using The Aerospace Corporation’s prototype ground stations located in El Segundo, California. The Rogue-alpha, beta IR sensor is a 1.4 micron band, 640x512 pixel, 28° field of view, InGaAs SWIR camera. It is accompanied by a panchromatic, 10-megapixel, 37° field of view visible context camera. Modes of sensor operation have included: 1) horizon-pointed imaging in all directions relative to the spacecraft orbit (fore, aft, port, and starboard) which is designed to maximize the imaged field of view, 2) point-and-stare imaging, 3) nadir-pointed, and 4) stereo fore-aft pointing using both spacecraft. All of these modes of operation are usually conducted in multi-frame collections at 1-20hz for dozens to thousands of frames. Highlights from the Rogue-alpha, beta sensor Earth remote sensing observation experiments will be presented. These have included impressive video imagery of hurricanes, typhoons, thunderstorms, and high clouds in the intra-tropical convergence zone. Infrared and visible point sources studied include gas flares, wildfires, active volcanos, nighttime lights, and other phenomena, including the first infrared CubeSat observations of space launch upper stages in flight. Stereo cloud imaging observations were also conducted with an aim of better understanding Earth backgrounds from low Earth orbit. Highlights from the CubeSat flight operations experiments include: 1) spacecraft-to-spacecraft boresight alignment of Rogue’s lasercom systems, and 2) metric and radiometric calibration of Rogue’s flight cameras using bright infrared stars. The results from the Rogue-alpha, beta460-km orbit show the exciting possibilities for wide-field-of-view missions from low earth orbit

    Digital archiving and retrieval tool

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    Our poster will present DART, the Digital Archiving and Retrieval Tool, a system intended to support engineers in archiving digital data developed over the course of the design process. DART is a direct product of recent experiences in adding collaboration, archiving, and version control systems to project components of university-level engineering curricula. The primary insights include the necessity of minimal disruption to the workflow and the utility of a basic, data-grabbing, "cyber-forensics" approach to digital archiving. The system incorporates those principles, providing a tool by which useful data and context may be stored while imposing minimal learning curve or workflow change. An explicit tradeoff is made between the ability to easily manage and analyze archived data versus adoption of the tool and successful data archiving

    Sustained expansion of NKT cells and antigen-specific T cells after injection of α-galactosyl-ceramide loaded mature dendritic cells in cancer patients

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    Natural killer T (NKT) cells are distinct glycolipid reactive innate lymphocytes that are implicated in the resistance to pathogens and tumors. Earlier attempts to mobilize NKT cells, specifically, in vivo in humans met with limited success. Here, we evaluated intravenous injection of monocyte-derived mature DCs that were loaded with a synthetic NKT cell ligand, α-galactosyl-ceramide (α-GalCer; KRN-7000) in five patients who had advanced cancer. Injection of α-GalCer–pulsed, but not unpulsed, dendritic cells (DCs) led to >100-fold expansion of several subsets of NKT cells in all patients; these could be detected for up to 6 mo after vaccination. NKT activation was associated with an increase in serum levels of interleukin-12 p40 and IFN-γ inducible protein-10. In addition, there was an increase in memory CD8(+) T cells specific for cytomegalovirus in vivo in response to α-GalCer–loaded DCs, but not unpulsed DCs. These data demonstrate the feasibility of sustained expansion of NKT cells in vivo in humans, including patients who have advanced cancer, and suggest that NKT activation might help to boost adaptive T cell immunity in vivo

    Systematic Identification of Genes that Regulate Neuronal Wiring in the Drosophila Visual System

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    Forward genetic screens in model organisms are an attractive means to identify those genes involved in any complex biological process, including neural circuit assembly. Although mutagenesis screens are readily performed to saturation, gene identification rarely is, being limited by the considerable effort generally required for positional cloning. Here, we apply a systematic positional cloning strategy to identify many of the genes required for neuronal wiring in the Drosophila visual system. From a large-scale forward genetic screen selecting for visual system wiring defects with a normal retinal pattern, we recovered 122 mutations in 42 genetic loci. For 6 of these loci, the underlying genetic lesions were previously identified using traditional methods. Using SNP-based mapping approaches, we have now identified 30 additional genes. Neuronal phenotypes have not previously been reported for 20 of these genes, and no mutant phenotype has been previously described for 5 genes. The genes encode a variety of proteins implicated in cellular processes such as gene regulation, cytoskeletal dynamics, axonal transport, and cell signalling. We conducted a comprehensive phenotypic analysis of 35 genes, scoring wiring defects according to 33 criteria. This work demonstrates the feasibility of combining large-scale gene identification with large-scale mutagenesis in Drosophila, and provides a comprehensive overview of the molecular mechanisms that regulate visual system wiring
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