Factors affecting replication and cross-species transmission of feline immunodeficiency virus

In order to successfully invade a new species, lentiviruses must overcome restriction factors, dominant blocks to replication, and be able to make use of available host factors such as entry receptors for replication. Human immunodeficiency virus (HIV-1) infection is blocked at a post-entry stage by rhesus macaque TRIM5α, an effect that is enhanced by host factor cyclophilin A (CypA), but largely evades restriction by the human TRIM5α variant. HIV-1 replication is also inhibited by simian APOBEC3G but is able to evade restriction by human APOBEC3G by inducing its degradation through expression of accessory protein Vif. Viral entry and tissue tropism are determined by an interaction of the viral Env glycoprotein with a cell surface receptor and a seven transmembrane domain co-receptor. The feline immunodeficiency virus (FIV) infects diverse felid species including the African lion, where infection has likely been endemic since at least the late Pleistocene, and the domestic cat, a more recent host. For domestic cat strains of FIV (FIV-Fca), entry is mediated by host proteins CD134 and CXCR4, but the identity of receptors in non-domestic strains of FIV is unknown. This thesis demonstrates that two strains of FIV isolated from lions (FIV-Ple subtypes B and E) differ in their receptor tropism and that subtype E shares entry receptors with FIV-Fca. The findings suggest that alternative receptor usage is a strategy employed by FIV in this species and has implications for the disputed pathology and tissue tropism of infection in African lions. Next, we tested the hypothesis that species which have harboured lentiviral infection for a long time are better able to prevent viral replication than recent hosts. Whilst we found that TRIM5α is non-functional in all felid species tested, evidence of potent APOBEC3 activity was found and, in lion cells, potently restricts production of infectious FIV. Moreover, lion primary T-cells prevent replication of diverse FIV strains and restrict primate lentiviruses at post-entry stages, suggesting that co-evolution with lentiviruses has driven the selection of broad-ranging restriction factors. Structural and biophysical analyses suggest that whilst FIV’s interaction with CypA appears to be conserved in affinity with HIV-1, the interaction does not appear to be crucial for replication and, in the absence of restriction by TRIM5α, the role of the capsid-CypA interaction is discussed. Overall the study explains the permissivity of domestic cat cells to retroviral infection and identifies FIV infection of lions as an example of host adaptation driven by current lentiviral infection

A digital video system for observing and recording occultations

Stellar occultations by asteroids and outer solar system bodies can offer ground based observers with modest telescopes and camera equipment the opportunity to probe the shape, size, atmosphere and attendant moons or rings of these distant objects. The essential requirements of the camera and recording equipment are: good quantum efficiency and low noise, minimal dead time between images, good horological faithfulness of the image time stamps, robustness of the recording to unexpected failure, and low cost. We describe the Astronomical Digital Video occultation observing and recording System (ADVS) which attempts to fulfil these requirements and compare the system with other reported camera and recorder systems. Five systems have been built, deployed and tested over the past three years, and we report on three representative occultation observations: one being a 9 +/-1.5 second occultation of the trans-Neptunian object 28978 Ixion (mv=15.2) at 3 seconds per frame, one being a 1.51 +/-0.017 second occultation of Deimos, the 12~km diameter satellite of Mars, at 30 frames per second, and one being a 11.04 +/-0.4 second occultation, recorded at 7.5 frames per second, of the main belt asteroid, 361 Havnia, representing a low magnitude drop (Dmv = 0.4) occultation.Comment: 9 pages, 5 figures, 3 tables, accepted to Publications of the Astronomical Society of Australia (PASA

Design & Analysis of a Bicycle Towing Device

A local businessman had the idea to build a device that allows a bicycle to tow another bicycle. He built a prototype to demonstrate his idea but he wanted help in developing a product that could fit more bikes, did not require a rear rack to operate, and be manufacturable at his facility. He also expressed interest in an economic evaluation of production and marketing of the device. An analysis of customer needs led to adding reduction of weight and ease of attachment to the design requirements. This report is about improving the original design and presents the process of developing a working prototype that provides significant improvement to the original design. It discusses research regarding patents of similar products, bike frame strength, reliability and failure testing, and manufacturing methods. The design of each prototype is modeled in Solidworks and an analysis is performed with its built-in simulation software. The project resulted in a working prototype that allows a bicycle to tow another bicycle. The device requires no tools for installment. It uses wing head bolts to attach at the rear wheel hub, and a p-clamp to attach to the seatpost of the towing bike. The towed bike’s front wheel sits on a bar with hooked ends and the top of the wheel is tied to the device. It doesn’t require a luggage rack on the towing bike. We also included a description of the manufacturing methods and processing required to manufacture this device. The cost of the final design consists of material and labor costs. A facility layout is discussed with regard to production capacity

Truncation of TRIM5 in the <i>Feliformia</i> explains the absence of retroviral restriction in cells of the domestic cat

TRIM5[alpha] mediates a potent retroviral restriction phenotype in diverse mammalian species. Here, we identify a TRIM5 transcript in cat cells with a truncated B30.2 capsid binding domain and ablated restrictive function which, remarkably, is conserved across the &lt;i&gt;Feliformia&lt;/i&gt;. Cat TRIM5 displayed no restriction activity, but ectopic expression conferred a dominant negative effect against human TRIM5[alpha]. Our findings explain the absence of retroviral restriction in cat cells and suggest that disruption of the TRIM5 locus has arisen independently at least twice in the &lt;i&gt;Carnivora&lt;/i&gt;, with implications concerning the evolution of the host and pathogen in this taxon

Identification of cis-acting determinants mediating the unconventional secretion of tau.

The deposition of tau aggregates throughout the brain is a pathological characteristic within a group of neurodegenerative diseases collectively termed tauopathies, which includes Alzheimer's disease. While recent findings suggest the involvement of unconventional secretory pathways driving tau into the extracellular space and mediating the propagation of the disease-associated pathology, many of the mechanistic details governing this process remain elusive. In the current study, we provide an in-depth characterization of the unconventional secretory pathway of tau and identify novel molecular determinants that are required for this process. Here, using Drosophila models of tauopathy, we correlate the hyperphosphorylation and aggregation state of tau with the disease-related neurotoxicity. These newly established systems recapitulate all the previously identified hallmarks of tau secretion, including the contribution of tau hyperphosphorylation as well as the requirement for PI(4,5)P2 triggering the direct translocation of tau. Using a series of cellular assays, we demonstrate that both the sulfated proteoglycans on the cell surface and the correct orientation of the protein at the inner plasma membrane leaflet are critical determinants of this process. Finally, we identify two cysteine residues within the microtubule binding repeat domain as novel cis-elements that are important for both unconventional secretion and trans-cellular propagation of tau

Restriction of the felid lentiviruses by a synthetic feline TRIM5-CypA fusion

Gene therapy approaches to the treatment of HIV infection have targeted both viral gene expression and the cellular factors that are essential for virus replication. However, significant concerns have been raised regarding the potential toxic effects of such therapies, the emergence of resistant viral variants and unforeseen biological consequences such as enhanced susceptibility to unrelated pathogens. Novel restriction factors formed by the fusion of the tripartite motif protein (TRIM5) and cyclophilin A (CypA), or "TRIMCyps", offer an effective antiviral defence strategy with a very low potential for toxicity. In order to investigate the potential therapeutic utility of TRIMCyps in gene therapy for AIDS, a synthetic fusion protein between feline TRIM5 and feline CypA was generated and transduced into cells susceptible to infection with feline immunodeficiency virus (FIV). The synthetic feline TRIMCyp was highly efficient at preventing infection with both HIV and FIV and the cells resisted productive infection with FIV from either the domestic cat or the puma. Feline TRIMCyp and FIV infection of the cat offers a unique opportunity to evaluate TRIMCyp-based approaches to genetic therapy for HIV infection and the treatment of AIDS