A novel method to titrate Herpes simplex virus-1 (HSV-1) using laser-based scanning of near-infrared fluorophores conjugated antibodies

Among several strategies used for Herpes simplex virus (HSV) detection in biological specimens, standard plaque assay (SPA) remains the most reliable method to evaluate virus infectivity and quantify viral replication. However, it is a manual procedure, thereby affected by operator subjectivity, and it may be particularly laborious for multiple sample analysis. Here we describe an innovative method to perform the titration of HSV type 1 (HSV-1) in different samples, using the “In-Cell WesternTM” Assay (ICW) from LI-COR, a quantitative immunofluorescence assay that exploits laser-based scanning of near infrared (NIR). In particular, we employed NIR-immunodetection of viral proteins to monitor foci of HSV-1 infection in cell monolayers, and exploited an automated detection of their fluorescence intensity to evaluate virus titre. This innovative method produced similar and superimposable values compared to SPA, but it is faster and can be performed in 96 well plate, thus allowing to easily and quickly analyze and quantify many samples in parallel. These features make our method particularly suitable for the screening and characterization of antiviral compounds, as we demonstrated by testing acyclovir (ACV), the main anti-HSV-1 drug. Moreover, we developed a new data analysis system that allowed to overcome potential bias due to unspecific florescence signals, thus improving data reproducibility. Overall, our method may represents a useful tool for both clinical and research purposes

Conception d'une base recueillant des données agronomiques

La culture de la canne à sucre est un véritable enjeu économique, social et environnemental sur l'île de La Réunion. La surface plantée en canne à sucre s'estime à 26% dans les " Hauts ", où les températures sont les plus basses, engendrant de nombreuses contraintes culturales. C'est pourquoi le Cirad réalise des essais sur des variétés nouvellement adaptées à ces contraintes de basses températures. Ces essais se répartissent sur quatre stations : Bassin Plat, La Mare, Ligne Paradis et les Colimaçons et génèrent un flot important de données agronomiques, actuellement enregistrées dans des feuilles de calculs Excel. Les données collectées contribuent par la suite à des études statistiques. Les chercheurs du Cirad des pôles Sud et Nord de l'île souhaitent mettre en commun leurs données dans une base, exploitable via le Web. Une structure de cette base a été proposée après un travail de réflexion préalable reposant sur la conception de modèles. (Résumé d'auteur

Herpes simplex virus-type1 (HSV-1) impairs DNA repair in cortical neurons

Several findings suggest that Herpes simplex virus-1 (HSV-1) infection plays a role in the neurodegenerative processes that characterize Alzheimer's disease (AD), but the underlying mechanisms have yet to be fully elucidated. Here we show that HSV-1 productive infection in cortical neurons causes the accumulation of DNA lesions that include both single (SSBs) and double strand breaks (DSBs), which are reported to be implicated in the neuronal loss observed in neurodegenerative diseases. We demonstrate that HSV-1 downregulates the expression level of Ku80, one of the main components of non-homologous end joining (NHEJ), a major pathway for the repair of DSBs. We also provide data suggesting that HSV-1 drives Ku80 for proteasomal degradation and impairs NHEJ activity, leading to DSB accumulation. Since HSV-1 usually causes life-long recurrent infections, it is possible to speculate that cumulating damages, including those occurring on DNA, may contribute to virus induced neurotoxicity and neurodegeneration, further suggesting HSV-1 as a risk factor for neurodegenerative conditions

Effects of vertical vibration on hopper flows of granular material

The discharge of granular material from a hopper subject to vertical sinusoidal oscillations was investigated using experiments and discrete element computer simulations. With the hopper exit closed, side-wall convection cells are observed, oriented such that particles move up along the inclined walls of the hopper and down at the center line. The convection cells are a result of the granular bed dilation during free fall and the subsequent interaction with the hopper walls. The mass discharge rate for a vibrating hopper scaled by the discharge rate without vibration reaches a maximum value at a dimensionless velocity amplitude just greater than 1. Further increases in the velocity decrease the discharge rate. The decrease occurs due to a decrease in the bulk density of the discharging material when vibration is applied

Role of glutathionylation in infection and inflammation

Glutathionylation, that is, the formation of mixed disulfides between protein cysteines and glutathione (GSH) cysteines, is a reversible post-translational modification catalyzed by dierent cellular oxidoreductases, by which the redox state of the cell modulates protein function. So far, most studies on the identification of glutathionylated proteins have focused on cellular proteins, including proteins involved in host response to infection, but there is a growing number of reports showing that microbial proteins also undergo glutathionylation, with modification of their characteristics and functions. In the present review, we highlight the signaling role of GSH through glutathionylation, particularly focusing on microbial (viral and bacterial) glutathionylated proteins (GSSPs) and host GSSPs involved in the immune/inflammatory response to infection; moreover, we discuss the biological role of the process in microbial infections and related host responses

The variance of identity-by-descent sharing in the Wright-Fisher model

Widespread sharing of long, identical-by-descent (IBD) genetic segments is a hallmark of populations that have experienced recent genetic drift. Detection of these IBD segments has recently become feasible, enabling a wide range of applications from phasing and imputation to demographic inference. Here, we study the distribution of IBD sharing in the Wright-Fisher model. Specifically, using coalescent theory, we calculate the variance of the total sharing between random pairs of individuals. We then investigate the cohort-averaged sharing: the average total sharing between one individual and the rest of the cohort. We find that for large cohorts, the cohort-averaged sharing is distributed approximately normally. Surprisingly, the variance of this distribution does not vanish even for large cohorts, implying the existence of "hyper-sharing" individuals. The presence of such individuals has consequences for the design of sequencing studies, since, if they are selected for whole-genome sequencing, a larger fraction of the cohort can be subsequently imputed. We calculate the expected gain in power of imputation by IBD, and subsequently, in power to detect an association, when individuals are either randomly selected or specifically chosen to be the hyper-sharing individuals. Using our framework, we also compute the variance of an estimator of the population size that is based on the mean IBD sharing and the variance in the sharing between inbred siblings. Finally, we study IBD sharing in an admixture pulse model, and show that in the Ashkenazi Jewish population the admixture fraction is correlated with the cohort-averaged sharing.Comment: Includes Supplementary Materia

Geomagnetic activity forcing of the Northern Annular Mode via the stratosphere

We consider various aspects of the link between solar-modulated geomagnetic activity and the Northern Annular Mode (NAM). Our results indicate that the geomagnetic forcing of atmospheric circulation in the Northern Hemisphere is temporally and seasonally restricted, modulated by the Quasi-Biennial Oscillation (QBO), and reliant on stratosphere-troposphere coupling. When the data are restricted to January values after 1965, for years in which the January QBO is eastwards, the correlation coefficient between the geomagnetic AA index and the NAM is 0.85. These results can account for many of the enigmatic features of Northern Hemisphere circulation.<br><br> <b>Key words.</b> Meterology and atmospheric dynamics (general circulation, climatology

EFFICIENT PROPAGATION OF ARCHETYPE JC POLYOMAVIRUS IN COS-7 CELLS: EVALUATION OF REARRANGEMENTS WITHIN NCCR STRUCTURAL ORGANIZATION DURING TRANSFECTION.

John Cunningham virus (JCPyV) is an ubiqui-tous human pathogen that causes disease in immunocom-promised patients. The JCPyV genome is composed of an early region and a late region, which are physically sepa-rated by the non-coding control region (NCCR). The DNA sequence of the NCCR distinguishes two forms of JCPyV, the designated archetype and the prototype, which resulted from a rearrangement of the archetype sequence. To date, the cell culture systems for propagating JCPyV archetype have been very limited in their availability and robust-ness. Prior to this study, it was demonstrated that JCPyV archetype DNA replicates in COS-7 simian kidney cells expressing SV40 TAg and COS-7 cells expressing HIV-1 Tat. Based on these observations, the present study was conducted to reproduce an in vitro model in COS-7 cells transfected with the JCPyV archetype strain in order to study JCPyV DNA replication and analyze NCCR rear-rangements during the viral life cycle. The efficiency of JCPyV replication was evaluated by quantitative PCR (Q-PCR) and by hemagglutination (HA) assay after trans-fection. In parallel, sequence analysis of JCPyV NCCR was performed. JCPyV efficiently replicated in kidney-derived COS-7 cells, as demonstrated by a progressive increase in viral load and virion particle production after transfection. The archetypal structure of NCCR was maintained during the viral cycle, but two characteristic point mutations were detected 28 days after transfection. This model is a useful tool for analyzing NCCR rearrangements during in vitroreplication in cells that are sites of viral persistence, such as tubular epithelial cells of the kidne