Creating a Virtual Ethnographic Field School in an Off-line Community of Practice

This paper describes the creation of an asynchronous on-line ethnographic field school experience for lower division undergraduate students. Our Virtual Field School course offers a field school experience that accommodates the unique make-up of the University of Alaska Fairbanks (where fifty-five percent of undergraduates are “nontraditional” students). Typical ethnographic field schools demand that students can spend four to six weeks in an international fieldsite. Alaska’s geographic remoteness makes travel abroad prohibitively expensive for many students. Pedagogical and technological concerns are outlined, including the utilization of the SELIN distance delivery platform, coupled with Blackboard Learn. SELIN was created by anthropologists at the University of Neuchâtel (Switzerland) as a means of teaching upper division anthropology majors inductive reasoning and observation skills. SELIN courses are centered on authentic multimedia documentation of fieldsites, rather than texts or lectures. The pedagogical merits of the choice of the ethnographic site—the Alaska dog mushing community of practice—are discussed in light of novice anthropology students’ interests and abilities. The paper concludes by discussing the potential appeal of the virtual field school model to archaeology and the value of the unique type of multimedia materials created for the course for educational outreach

Selective SLAM/CD150 Receptor-Detargeting of Canine Distemper Virus.

The envelope attachment (H)-protein of canine distemper virus (CDV) mediates receptor engagement and fusion-triggering; two key functions in viral cell entry and spread. Signaling lymphocyte activation molecule (SLAM) and Nectin-4 (N4) act as morbilliviral entry receptors in immune and epithelial cells, respectively, which defines very similar pathogeneses. High incidence of brain disorders is however unique to CDV. The wild-type CDV-A75/17 strain (A75) preferentially infects glial cells and spreads from astrocyte-to-astrocyte without inducing massive fusion events, despite the fact that SLAM and N4 expressions remained below detection levels. To investigate whether an A75 H-microdomain required to interact with SLAM may additionally contribute to promote viral spread between astrocytes, we initially engineered a novel A75 H-protein variant (546-SYT/RNR-548) that lost SLAM-binding property and, consequently, lacked fusion-triggering activity specifically in SLAM-expressing cells. Collectively, this approach provides the molecular tool to decipher the role of the selected H-microdomain in supporting A75-spread in glial cells

“Neighborhood in Solidarity”: A community development methodology that emerged throughout an action research experience

Population aging and urbanization are often associated with a restriction of the living environment and an increasing tendency to remain at home. This community development report presents the ‘‘Neighborhood in Solidarity’’ methodology as a response to counter or at least slow this restriction and enhance the ability of elderly persons to be active within their neighborhoods. Co-constructed throughout the action research process, the Neighborhood in Solidarity methodology is based on a 13-year experience accumulated through 22 projects in 17 cities, with promising results in the Canton of Vaud in Switzerland. The genesis and the emergence of the methodology throughout action research workshops and interregional structures are described in the document. The description of Neighborhood in Solidarity is a snapshot of a living methodology, which continues to evolve on a daily basis. The process focuses on empowering the older people through a five-year methodology, which is intended to create an autonomous community that can resolve its own problems. The methodology comprises six steps described throughout the paper: (1) preliminary analysis, (2) diagnostic, (3) construction, (4) project design, (5) project implementation, and (6) empowerment. In 2013, an external assessment evaluated the Neighborhood in Solidarity methodology as effective at and appropriate for achieving its objectives. The promising results of this original methodology motivated this publication

Sick of Shamanizing

Dans la Fédération de Russie, les phénomènes de mobilité à motivation religieuse ont connu une croissance rapide dès les premières heures de la période postsoviétique. Au Kamtchatka (Extrême-Orient russe), le principal moteur de ces déplacements est un mouvement missionnaire d’inspiration pentecôtiste parti d’Ukraine à la fin des années 1980. Ces nouveaux circuits évangéliques ne sont pas dépourvus de formes indigènes de “chamanisation”. Certains attributs phares, comme jouer du tambour, ont voyagé depuis les confins du Nord-Kamtchatka jusque dans les églises urbaines du mouvement et sont devenus charismatiques. À l’inverse, la fabrication “d’idoles” zoomorphes véhicule l’illustration d’une maladie grave dont il faut guérir. Dans cet article, je me penche sur les modalités d’incorporation et de mise à distance du jeu de tambour et de la vénération d’idoles sur les nouvelles routes de la foi ouvertes au Kamtchatka postsoviétique. J’examine la façon dont chamaniser constitue à la fois un remède aux maux contemporains et une pathologie d’un autre temps. Je suggère que cette perspective alambiquée brouille la définition, déjà floue, du chamanisme au Kamtchatka et permet à une poignée de pentecôtistes de prendre la route.In the Russian Federation, religiously motivated mobility phenomena have grown rapidly in the post-Soviet aftermath. In Kamchatka (Russian Far East), the main driving force behind religious mobility is a neo-Pentecostal missionary movement that spread from Ukraine in the late 1980s. Native forms of “shamanizing” are not absent from this new Christian world. Some of their emblematic features, such as drumming, travelled all the way from Northern Kamchatka to the urban churches of the movement where they became charismatic. By contrast, the making of zoomorphic “idols” conveys the idea of a sickness that requires appropriate religious healing. In this paper, I examine how drumming and idol worshipping are respectively carried away and left behind on the new roads of faith opened in post-Soviet Kamchatka. I look at how shamanizing can be at the same time a remedy against post-Soviet harms, and a pathology from another time. I suggest that this twisted view blurs the (already blurry) definition of shamanism in Kamchatka and keeps a handful of Pentecostals on the move

Canine distemper virus persistence in demyelinating encephalitis by swift intracellular cell-to-cell spread in astrocytes is controlled by the viral attachment protein

The mechanism of viral persistence, the driving force behind the chronic progression of inflammatory demyelination in canine distemper virus (CDV) infection, is associated with non-cytolytic viral cell-to-cell spread. Here, we studied the molecular mechanisms of viral spread of a recombinant fluorescent protein-expressing virulent CDV in primary canine astrocyte cultures. Time-lapse video microscopy documented that CDV spread was very efficient using cell processes contacting remote target cells. Strikingly, CDV transmission to remote cells could occur in less than 6h, suggesting that a complete viral cycle with production of extracellular free particles was not essential in enabling CDV to spread in glial cells. Titration experiments and electron microscopy confirmed a very low CDV particle production despite higher titers of membrane-associated viruses. Interestingly, confocal laser microscopy and lentivirus transduction indicated expression and functionality of the viral fusion machinery, consisting of the viral fusion (F) and attachment (H) glycoproteins, at the cell surface. Importantly, using a single-cycle infectious recombinant H-knockout, H-complemented virus, we demonstrated that H, and thus potentially the viral fusion complex, was necessary to enable CDV spread. Furthermore, since we could not detect CD150/SLAM expression in brain cells, the presence of a yet non-identified glial receptor for CDV was suggested. Altogether, our findings indicate that persistence in CDV infection results from intracellular cell-to-cell transmission requiring the CDV-H protein. Viral transfer, happening selectively at the tip of astrocytic processes, may help the virus to cover long distances in the astroglial network, "outrunning” the host's immune response in demyelinating plaques, thus continuously eliciting new lesion

Efficient Recovery of Attenuated Canine Distemper Virus from cDNA.

To provide insights into the biology of the attenuated canine distemper virus (CDV) Onderstepoort (OP) strain (large plaque forming variant), design next-generation multivalent vaccines, or further investigate its promising potential as an oncolytic vector, we employed contemporary modifications to establish an efficient OP-CDV-based reverse genetics platform. Successful viral rescue was obtained however only upon recovery of a completely conserved charged residue (V13E) residing at the N-terminal region of the large protein (L). Although L-V13 and L-V13E did not display drastic differences in cellular localization and physical interaction with P, efficient polymerase complex (P+L) activity was recorded only with L-V13E. Interestingly, grafting mNeonGreen to the viral N protein via a P2A ribosomal skipping sequence (OPneon) and its derivative V-protein-knockout variant (OPneon-Vko) exhibited delayed replication kinetics in cultured cells. Collectively, we established an efficient OP-CDV-based reverse genetics system that enables the design of various strategies potentially contributing to veterinary medicine and research

Oligomerization and Cell Egress Controlled by Two Microdomains of Canine Distemper Virus Matrix Protein.

The multimeric matrix (M) protein of clinically relevant paramyxoviruses orchestrates assembly and budding activity of viral particles at the plasma membrane (PM). We identified within the canine distemper virus (CDV) M protein two microdomains, potentially assuming α-helix structures, which are essential for membrane budding activity. Remarkably, while two rationally designed microdomain M mutants (E89R, microdomain 1 and L239D, microdomain 2) preserved proper folding, dimerization, interaction with the nucleocapsid protein, localization at and deformation of the PM, the virus-like particle formation, as well as production of infectious virions (as monitored using a membrane budding-complementation system), were, in sharp contrast, strongly impaired. Of major importance, raster image correlation spectroscopy (RICS) revealed that both microdomains contributed to finely tune M protein mobility specifically at the PM. Collectively, our data highlighted the cornerstone membrane budding-priming activity of two spatially discrete M microdomains, potentially by coordinating the assembly of productive higher oligomers at the PM.IMPORTANCE Despite the availability of efficient vaccines, morbilliviruses (e.g., canine distemper virus [CDV] and measles virus [MeV]) still cause major health impairments. Although antivirals may support vaccination campaigns, approved inhibitors are to date still lacking. Targeting late stages of the viral life cycle (i.e., the cell exit system) represents a viable option to potentially counteract morbilliviral infections. The matrix (M) protein of morbillivirus is a major contributor to membrane budding activity and is assumed to assemble into dimers that further associate to form higher oligomers. Here, we rationally engineered M protein variants with modifications in two microdomains that potentially locate at dimer-dimer interfaces. Our results spotlight the cornerstone impact of both microdomains in membrane budding activity and further suggest a role of finely tuned high-order oligomer formation in regulating late stages of cell exit. Collectively, our findings highlight two microdomains in the morbilliviral M protein as novel attractive targets for drug design

Le double jeu de la chance: imitation et substitution dans les rituels chamaniques contemporains de deux populations rurales du Nord-Kamtchatka (Fédération de Russie, Extrême-Orient sibérien) : les chasseurs maritimes de Lesnaia et les éleveurs de rennes d'Atchaïvaiam

Le " Double jeu de la chance " est une ethnographie croisée des pratiques et représentations de la chasse au gros gibier et de l'élevage de rennes dans le Nord-Kamtchatka contemporain en vue de leur comparaison sous l'angle rituel. La première partie de la thèse vise à présenter les aspects de l'organisation sociale susceptibles d'éclairer les rituels. Elle porte sur les fondements de la mobilité post-soviétique de deux populations rurales contemporaines : les chasseurs maritimes de Lesnaia et les éleveurs de rennes d'Atchaïvaiam. La seconde partie de la thèse vise à mettre au jour les mécanismes de la mise en circulation de la " chance ", notion omniprésente en Sibérie et analysée comme " principe vital ou force de vie " (Hamayon). Deux modes privilégiés d'obtention de la chance émergent chez les Lesnovskiens et les Atchaïvaiamskiens : la réalisation de " jeux " rituels (uzizičvet) et les pratiques sacrificielles. Les premiers entretiennent un rapport étroit avec la notion d'imitation, alors que les secondes sont dominées par le principe de substitution (Lévi-Strauss). Examinées séparément puis en commun, ces deux catalyseurs de l'efficacité rituelle s'articulent pour permettre la (re)mise en mouvement(s) symbolique des proies abattues (phoques, ours, mouflons) ou des victimes sacrificielles (rennes). Une série d'exemples (courses de rennes, rituel de chasse ololo, rites funéraires) souligne l'importance des substituts dans le système de représentation chamanique des éleveurs et des chasseurs. In fine, la thèse suggère que la convergence des " jeux " et des sacrifices est indispensable aux transferts des " âmes " et à la mobilité générale du monde post-soviétique

LRP6 Is a Functional Receptor for Attenuated Canine Distemper Virus.

Wild-type canine distemper virus (CDV) is an important pathogen of dogs as well as wildlife that can infect immune and epithelial cells through two known receptors: the signaling lymphocytic activation molecule (SLAM) and nectin-4, respectively. Conversely, the ferret and egg-adapted CDV-Onderstepoort strain (CDV-OP) is employed as an effective vaccine for dogs. CDV-OP also exhibits promising oncolytic properties, such as its abilities to infect and kill multiple cancer cells in vitro. Interestingly, several cancer cells do not express SLAM or nectin-4, suggesting the presence of a yet unknown entry factor for CDV-OP. By conducting a genome-wide CRISPR/Cas9 knockout (KO) screen in CDV-OP-susceptible canine mammary carcinoma P114 cells, which neither express SLAM nor nectin-4, we identified low-density lipoprotein receptor-related protein 6 (LRP6) as a host factor that promotes CDV-OP infectivity. Whereas the genetic ablation of LRP6 rendered cells resistant to infection, ectopic expression in resistant LRP6KO cells restored susceptibility. Furthermore, multiple functional studies revealed that (i) the overexpression of LRP6 leads to increased cell-cell fusion, (ii) a soluble construct of the viral receptor-binding protein (solHOP) interacts with a soluble form of LRP6 (solLRP6), (iii) an H-OP point mutant that prevents interaction with solLRP6 abrogates cell entry in multiple cell lines once transferred into recombinant viral particles, and (iv) vesicular stomatitis virus (VSV) pseudotyped with CDV-OP envelope glycoproteins loses its infectivity in LRP6KO cells. Collectively, our study identified LRP6 as the long sought-after cell entry receptor of CDV-OP in multiple cell lines, which set the molecular bases to refine our understanding of viral-cell adaptation and to further investigate its oncolytic properties. IMPORTANCE Oncolytic viruses (OV) have gathered increasing interest in recent years as an alternative option to treat cancers. The Onderstepoort strain of canine distemper virus (CDV-OP), an enveloped RNA virus belonging to the genus Morbillivirus, is employed as a safe and efficient vaccine for dogs against distemper disease. Importantly, although CDV-OP can infect and kill multiple cancer cell lines, the basic mechanisms of entry remain to be elucidated, as most of those transformed cells do not express natural receptors (i.e., SLAM and nectin-4). In this study, using a genome-wide CRISPR/Cas9 knockout screen, we describe the discovery of LRP6 as a novel functional entry receptor for CDV-OP in various cancer cell lines and thereby uncover a basic mechanism of cell culture adaptation. Since LRP6 is upregulated in various cancer types, our data provide important insights in order to further investigate the oncolytic properties of CDV-OP

Antiviral Screen against Canine Distemper Virus-Induced Membrane Fusion Activity.

Canine distemper virus (CDV), a close relative of the human pathogen measles virus (MeV), is an enveloped, negative sense RNA virus that belongs to the genus Morbillivirus and causes severe diseases in dogs and other carnivores. Although the vaccination is available as a preventive measure against the disease, the occasional vaccination failure highlights the importance of therapeutic alternatives such as antivirals against CDV. The morbilliviral cell entry system relies on two interacting envelope glycoproteins: the attachment (H) and fusion (F) proteins. Here, to potentially discover novel entry inhibitors targeting CDV H, F and/or the cognate receptor: signaling lymphocyte activation molecule (SLAM) proteins, we designed a quantitative cell-based fusion assay that matched high-throughput screening (HTS) settings. By screening two libraries of small molecule compounds, we successfully identified two membrane fusion inhibitors (F2736-3056 and F2261-0043). Although both inhibitors exhibited similarities in structure and potency with the small molecule compound 3G (an AS-48 class morbilliviral F-protein inhibitor), F2736-3056 displayed improved efficacy in blocking fusion activity when a 3G-escape variant was employed. Altogether, we present a cell-based fusion assay that can be utilized not only to discover antiviral agents against CDV but also to dissect the mechanism of morbilliviral-mediated cell-binding and cell-to-cell fusion activity