Use of 5-(Perylen-3-yl)Ethynyl-arabino-Uridine (aUY11) to inactivate Rift Valley fever virus and preserve neutralization epitopes

Rift Valley fever (RFV) is a mosquito-borne disease, endemic to Africa, and known to cause abortion in animals and hemorrhagic fever in humans. All enveloped viruses require the fusion of viral membranes with cellular membranes to enter into and replicate inside the host cell. These viruses have a universal objective: find the cell receptors that match the viral attachment proteins and use those to gain access to the cell membrane. The use of aUY11, a monosaccharide (sugar)-based compound, has previously shown to effectively use a biophysical approach in the inhibition of viral fusion of other enveloped viruses1. In this study, we attempt to use aUY11 to inhibit the fusion of Rift Valley fever virus (RVFV) in vitro as a primary step in the creation of an FDA-approved vaccination for humans and animals to protect against often-lethal RVFV infection. We show here that aUY11 inhibits viral replication in Vero 76 cells and therefore has potential as an RVFV vaccine

The cumulative build-up of Pasteuria penetrans spores in root-knot nematode infested soil and the effect of soil applied fungicides on its infectivity

La croissance cumulée de spores de #Pasteuria penetrans dans le sol (en pots ou en champ) après trois cycles culturaux a été observée après ré-incorporation de racines de concombre ou de tomate infestées par #Meloidogyne spp. eux-mêmes parasités par #P. penetrans. Lors d'une expérience en pots, la ré-inoculation de racines de tomates infestés par #Meloidogyne spp. parasitées par #P. penetrans augmente le nombre de spores à un niveau tel que l'invasion par le nématode en est empêchée. Lors d'une expérience en champ, les juvéniles de deuxième stade extraits du sol après trois cycles de culture de concombre ne présentent que huit spores fixées en moyenne. Ni la fixation ni le développement du parasite ne sont affectés si les spores de #P. penetrans sont exposées à ces quatre fongicides courants. (Résumé d'auteur

Comparative study of different scattering geometries for the proposed Indian X-ray polarization measurement experiment using Geant4

Polarization measurements in X-rays can provide unique opportunity to study the behavior of matter and radiation under extreme magnetic fields and extreme gravitational fields. Unfortunately, over past two decades, when X-ray astronomy witnessed multiple order of magnitude improvement in temporal, spatial and spectral sensitivities, there is no (or very little) progress in the field of polarization measurements of astrophysical X-rays. Recently, a proposal has been submitted to ISRO for a dedicated small satellite based experiment to carry out X-ray polarization measurement, which aims to provide the first X-ray polarization measurements since 1976. This experiment will be based on the well known principle of polarization measurement by Thomson scattering and employs the baseline design of a central low Z scatterer surrounded by X-ray detectors to measure the angular intensity distribution of the scattered X-rays. The sensitivity of such experiment is determined by the collecting area, scattering and detection efficiency, X-ray detector background, and the modulation factor. Therefore, it is necessary to carefully select the scattering geometry which can provide the highest modulation factor and thus highest sensitivity within the specified experimental constraints. The effective way to determine optimum scattering geometry is by studying various possible scattering geometries by means of Monte Carlo simulations. Here we present results of our detailed comparative study based on Geant4 simulations of five different scattering geometries which can be considered within the weight and size constraints of the proposed small satellite based X-ray polarization measurement experiment.Comment: 14 pages, 6 figures, accepted for publication in "Nuclear Inst. and Methods in Physics Research, A

Genome-scale metabolic analysis of Clostridium thermocellum for bioethanol production

<p>Abstract</p> <p>Background</p> <p>Microorganisms possess diverse metabolic capabilities that can potentially be leveraged for efficient production of biofuels. <it>Clostridium thermocellum </it>(ATCC 27405) is a thermophilic anaerobe that is both cellulolytic and ethanologenic, meaning that it can directly use the plant sugar, cellulose, and biochemically convert it to ethanol. A major challenge in using microorganisms for chemical production is the need to modify the organism to increase production efficiency. The process of properly engineering an organism is typically arduous.</p> <p>Results</p> <p>Here we present a genome-scale model of <it>C. thermocellum </it>metabolism, <it>i</it>SR432, for the purpose of establishing a computational tool to study the metabolic network of <it>C. thermocellum </it>and facilitate efforts to engineer <it>C. thermocellum </it>for biofuel production. The model consists of 577 reactions involving 525 intracellular metabolites, 432 genes, and a proteomic-based representation of a cellulosome. The process of constructing this metabolic model led to suggested annotation refinements for 27 genes and identification of areas of metabolism requiring further study. The accuracy of the <it>i</it>SR432 model was tested using experimental growth and by-product secretion data for growth on cellobiose and fructose. Analysis using this model captures the relationship between the reduction-oxidation state of the cell and ethanol secretion and allowed for prediction of gene deletions and environmental conditions that would increase ethanol production.</p> <p>Conclusions</p> <p>By incorporating genomic sequence data, network topology, and experimental measurements of enzyme activities and metabolite fluxes, we have generated a model that is reasonably accurate at predicting the cellular phenotype of <it>C. thermocellum </it>and establish a strong foundation for rational strain design. In addition, we are able to draw some important conclusions regarding the underlying metabolic mechanisms for observed behaviors of <it>C. thermocellum </it>and highlight remaining gaps in the existing genome annotations.</p

Gap Detection for Genome-Scale Constraint-Based Models

Constraint-based metabolic models are currently the most comprehensive system-wide models of cellular metabolism. Several challenges arise when building an in silico constraint-based model of an organism that need to be addressed before flux balance analysis (FBA) can be applied for simulations. An algorithm called FBA-Gap is presented here that aids the construction of a working model based on plausible modifications to a given list of reactions that are known to occur in the organism. When applied to a working model, the algorithm gives a hypothesis concerning a minimal medium for sustaining the cell in culture. The utility of the algorithm is demonstrated in creating a new model organism and is applied to four existing working models for generating hypotheses about culture media. In modifying a partial metabolic reconstruction so that biomass may be produced using FBA, the proposed method is more efficient than a previously proposed method in that fewer new reactions are added to complete the model. The proposed method is also more accurate than other approaches in that only biologically plausible reactions and exchange reactions are used

Rift Valley Fever Virus Infection in Golden Syrian Hamsters

Rift Valley fever virus (RVFV) is a formidable pathogen that causes severe disease and abortion in a variety of livestock species and a range of disease in humans that includes hemorrhagic fever, fulminant hepatitis, encephalitis and blindness. The natural transmission cycle involves mosquito vectors, but exposure can also occur through contact with infected fluids and tissues. The lack of approved antiviral therapies and vaccines for human use underlies the importance of small animal models for proof-of-concept efficacy studies. Several mouse and rat models of RVFV infection have been well characterized and provide useful systems for the study of certain aspects of pathogenesis, as well as antiviral drug and vaccine development. However, certain host-directed therapeutics may not act on mouse or rat pathways. Here, we describe the natural history of disease in golden Syrian hamsters challenged subcutaneously with the pathogenic ZH501 strain of RVFV. Peracute disease resulted in rapid lethality within 2 to 3 days of RVFV challenge. High titer viremia and substantial viral loads were observed in most tissues examined; however, histopathology and immunostaining for RVFV antigen were largely restricted to the liver. Acute hepatocellular necrosis associated with a strong presence of viral antigen in the hepatocytes indicates that fulminant hepatitis is the likely cause of mortality. Further studies to assess the susceptibility and disease progression following respiratory route exposure are warranted. The use of the hamsters to model RVFV infection is suitable for early stage antiviral drug and vaccine development studies

Treatment of Late Stage Disease in a Model of Arenaviral Hemorrhagic Fever: T-705 Efficacy and Reduced Toxicity Suggests an Alternative to Ribavirin

A growing number of arenaviruses are known to cause viral hemorrhagic fever (HF), a severe and life-threatening syndrome characterized by fever, malaise, and increased vascular permeability. Ribavirin, the only licensed antiviral indicated for the treatment of certain arenaviral HFs, has had mixed success and significant toxicity. Since severe arenaviral infections initially do not present with distinguishing symptoms and are difficult to clinically diagnose at early stages, it is of utmost importance to identify antiviral therapies effective at later stages of infection. We have previously reported that T-705, a substituted pyrazine derivative currently under development as an anti-influenza drug, is highly active in hamsters infected with Pichinde virus when the drug is administered orally early during the course of infection. Here we demonstrate that T-705 offers significant protection against this lethal arenaviral infection in hamsters when treatment is begun after the animals are ill and the day before the animals begin to succumb to disease. Importantly, this coincides with the time when peak viral loads are present in most organs and considerable tissue damage is evident. We also show that T-705 is as effective as, and less toxic than, ribavirin, as infected T-705-treated hamsters on average maintain their weight better and recover more rapidly than animals treated with ribavirin. Further, there was no added benefit to combination therapy with T-705 and ribavirin. Finally, pharmacokinetic data indicate that plasma T-705 levels following oral administration are markedly reduced during the latter stages of disease, and may contribute to the reduced efficacy seen when treatment is withheld until day 7 of infection. Our findings support further pre-clinical development of T-705 for the treatment of severe arenaviral infections

Use of Recombinant Adenovirus Vectored Consensus IFN-α to Avert Severe Arenavirus Infection

Several arenaviruses can cause viral hemorrhagic fever, a severe disease with case-fatality rates in hospitalized individuals ranging from 15-30%. Because of limited prophylaxis and treatment options, new medical countermeasures are needed for these viruses classified by the National Institutes of Allergy and Infectious Diseases (NIAID) as top priority biodefense Category A pathogens. Recombinant consensus interferon alpha (cIFN-α) is a licensed protein with broad clinical appeal. However, while cIFN-α has great therapeutic value, its utility for biodefense applications is hindered by its short in vivo half-life, mode and frequency of administration, and costly production. To address these limitations, we describe the use of DEF201, a replication-deficient adenovirus vector that drives the expression of cIFN-α, for pre- and post-exposure prophylaxis of acute arenaviral infection modeled in hamsters. Intranasal administration of DEF201 24 h prior to challenge with Pichindé virus (PICV) was highly effective at protecting animals from mortality and preventing viral replication and liver-associated disease. A significant protective effect was still observed with a single dosing of DEF201 given two weeks prior to PICV challenge. DEF201 was also efficacious when administered as a treatment 24 to 48 h post-virus exposure. The protective effect of DEF201 was largely attributed to the expression of cIFN-α, as dosing with a control empty vector adenovirus did not protect hamsters from lethal PICV challenge. Effective countermeasures that are highly stable, easily administered, and elicit long lasting protective immunity are much needed for arena and other viral infections. The DEF201 technology has the potential to address all of these issues and may serve as a broad-spectrum antiviral to enhance host defense against a number of viral pathogens