Plasmodium vivax antigen discovery based on alpha-helical coiled coil protein motif.

Protein α-helical coiled coil structures that elicit antibody responses, which block critical functions of medically important microorganisms, represent a means for vaccine development. By using bioinformatics algorithms, a total of 50 antigens with α-helical coiled coil motifs orthologous to Plasmodium falciparum were identified in the P. vivax genome. The peptides identified in silico were chemically synthesized; circular dichroism studies indicated partial or high α-helical content. Antigenicity was evaluated using human sera samples from malaria-endemic areas of Colombia and Papua New Guinea. Eight of these fragments were selected and used to assess immunogenicity in BALB/c mice. ELISA assays indicated strong reactivity of serum samples from individuals residing in malaria-endemic regions and sera of immunized mice, with the α-helical coiled coil structures. In addition, ex vivo production of IFN-γ by murine mononuclear cells confirmed the immunogenicity of these structures and the presence of T-cell epitopes in the peptide sequences. Moreover, sera of mice immunized with four of the eight antigens recognized native proteins on blood-stage P. vivax parasites, and antigenic cross-reactivity with three of the peptides was observed when reacted with both the P. falciparum orthologous fragments and whole parasites. Results here point to the α-helical coiled coil peptides as possible P. vivax malaria vaccine candidates as were observed for P. falciparum. Fragments selected here warrant further study in humans and non-human primate models to assess their protective efficacy as single components or assembled as hybrid linear epitopes.

A Pneumocystis jirovecii pneumonia outbreak in a single kidney-transplant center: role of cytomegalovirus co-infection.

Pneumocystis jirovecii pneumonia (PCP) and cytomegalovirus (CMV) infection represent possible complications of medical immunosuppression. Between 2005 and 2010, non-human immunodeficiency virus (HIV) PCP patients admitted to a nephrology unit were analyzed for outcome, CMV comorbidity, and patient-to-patient contacts prior to PCP. In contrast to 2002-2004 (no cases) and 2008-2010 (10 cases), a PCP outbreak of 29 kidney-transplant recipients and one patient with anti-glomerular basement membrane disease occurred between 2005 and 2007. None of the patients were on PCP chemoprophylaxis. In four PCP patients, the genotyping data of bronchoalveolar lavage specimen showed an identical Pneumocystis strain. PCP cases had a higher incidence of CMV infection (12 of 30 PCP patients) and CMV disease (four patients) when compared to matched PCP-free controls (p < 0.05). Cotrimoxazole and, if applicable, ganciclovir were started 2.0 ± 4.0 days following admission, and immunosuppressive medication was reduced. In-hospital mortality was 10% and the three-year mortality was 20%. CMV co-infection did not affect mortality. CMV co-infection more frequently occurred during a cluster outbreak of non-HIV PCP in comparison to PCP-free controls. Here, CMV awareness and specific therapy of both CMV infection and PCP led to a comparatively favorable patient outcome. The role of patient isolation should be further investigated in incident non-HIV PCP.

Where sound position influences sound object representations: a 7-T fMRI study.

Evidence from human and non-human primate studies supports a dual-pathway model of audition, with partially segregated cortical networks for sound recognition and sound localisation, referred to as the What and Where processing streams. In normal subjects, these two networks overlap partially on the supra-temporal plane, suggesting that some early-stage auditory areas are involved in processing of either auditory feature alone or of both. Using high-resolution 7-T fMRI we have investigated the influence of positional information on sound object representations by comparing activation patterns to environmental sounds lateralised to the right or left ear. While unilaterally presented sounds induced bilateral activation, small clusters in specific non-primary auditory areas were significantly more activated by contra-laterally presented stimuli. Comparison of these data with histologically identified non-primary auditory areas suggests that the coding of sound objects within early-stage auditory areas lateral and posterior to primary auditory cortex AI is modulated by the position of the sound, while that within anterior areas is not.

Multisensory anatomical pathways.

In order to interact with the multisensory world that surrounds us, we must integrate various sources of sensory information (vision, hearing, touch...). A fundamental question is thus how the brain integrates the separate elements of an object defined by several sensory components to form a unified percept. The superior colliculus was the main model for studying multisensory integration. At the cortical level, until recently, multisensory integration appeared to be a characteristic attributed to high-level association regions. First, we describe recently observed direct cortico-cortical connections between different sensory cortical areas in the non-human primate and discuss the potential role of these connections. Then, we show that the projections between different sensory and motor cortical areas and the thalamus enabled us to highlight the existence of thalamic nuclei that, by their connections, may represent an alternative pathway for information transfer between different sensory and/or motor cortical areas. The thalamus is in position to allow a faster transfer and even an integration of information across modalities. Finally, we discuss the role of these non-specific connections regarding behavioral evidence in the monkey and recent electrophysiological evidence in the primary cortical sensory areas.

RNA/DNA co-analysis from blood stains--results of a second collaborative EDNAP exercise.

A second collaborative exercise on RNA/DNA co-analysis for body fluid identification and STR profiling was organized by the European DNA Profiling Group (EDNAP). Six human blood stains, two blood dilution series (5-0.001 μl blood) and, optionally, bona fide or mock casework samples of human or non-human origin were analyzed by the participating laboratories using a RNA/DNA co-extraction or solely RNA extraction method. Two novel mRNA multiplexes were used for the identification of blood: a highly sensitive duplex (HBA, HBB) and a moderately sensitive pentaplex (ALAS2, CD3G, ANK1, SPTB and PBGD). The laboratories used different chemistries and instrumentation. All of the 18 participating laboratories were able to successfully isolate and detect mRNA in dried blood stains. Thirteen laboratories simultaneously extracted RNA and DNA from individual stains and were able to utilize mRNA profiling to confirm the presence of blood and to obtain autosomal STR profiles from the blood stain donors. The positive identification of blood and good quality DNA profiles were also obtained from old and compromised casework samples. The method proved to be reproducible and sensitive using different analysis strategies. The results of this collaborative exercise involving a RNA/DNA co-extraction strategy support the potential use of an mRNA based system for the identification of blood in forensic casework that is compatible with current DNA analysis methodology.

Aerosol immunization with NYVAC and MVA vectored vaccines is safe, simple, and immunogenic.

Each year, approximately five million people die worldwide from putatively vaccine-preventable mucosally transmitted diseases. With respect to mass vaccination campaigns, one strategy to cope with this formidable challenge is aerosol vaccine delivery, which offers potential safety, logistical, and cost-saving advantages over traditional vaccination routes. Additionally, aerosol vaccination may elicit pivotal mucosal immune responses that could contain or eliminate mucosally transmitted pathogens in a preventative or therapeutic vaccine context. In this current preclinical non-human primate investigation, we demonstrate the feasibility of aerosol vaccination with the recombinant poxvirus-based vaccine vectors NYVAC and MVA. Real-time in vivo scintigraphy experiments with radiolabeled, aerosol-administered NYVAC-C (Clade C, HIV-1 vaccine) and MVA-HPV vaccines revealed consistent mucosal delivery to the respiratory tract. Furthermore, aerosol delivery of the vaccines was safe, inducing no vaccine-associated pathology, in particular in the brain and lungs, and was immunogenic. Administration of a DNA-C/NYVAC-C prime/boost regime resulted in both systemic and anal-genital HIV-specific immune responses that were still detectable 5 months after immunization. Thus, aerosol vaccination with NYVAC and MVA vectored vaccines constitutes a tool for large-scale vaccine efforts against mucosally transmitted pathogens.

3

What is a virtual person? What is it used for? What is its added value? Virtual persons sometimes describe avatars and new forms of identities in online games. They also appear in other contexts; some authors use them in the legal domain. Within FIDIS, the concept of virtual person has been extended in order to better describe and understand new forms of identities in the Information Society in relation to rights, duties, obligations and responsibilities. Virtual persons, as other virtual entities, exist in the virtual world, the collection of all (abstract) entities which are or have been the product of the mind or imagination. The virtual world -not to be confused with the digital world- allows a unified description of many identity-related concepts that are usually defined separately without taking into consideration their similarities: avatars, pseudonyms, categories, profiles, legal persons, etc. The legal system has a long experience of using abstract entities to define rules, categories, etc., in order to associate legal rights, obligations, and responsibilities to persons that can be considered instances of these abstract entities in specific situations. The model developed within FIDIS intentionally uses a similar construction. n this chapter, after having explained the model, we apply it to pseudonyms. Then we explore the concept of virtual persons from a legal perspective. Eventually, we introduce trust in the light of virtual persons.

Inhimillisen kulttuurin reunaehtoja tutkimassa

Non-Human in Anthropology. Monitieteinen konferenssi 4.-5.12.2010 Prahassa.

Protocol for collection and separation of bone marrow mononuclear cells in Chlorocebus aethiops

Abstract: Chlorocebus aethiops is a species of non-human primate frequently used in biomedical research. Some research involves this species as an experimental model for various diseases and possible treatment with stem cells. The bone marrow is one of the main sources of these cells and provides easy access. The aim of this study was to standardize the protocol of collection and separation of bone marrow in C. aethiops. Ten animals were submitted to puncture of bone marrow with access to the iliac crest and cell separation by density gradient. The bone marrow of C. aethiops had an average of 97% viability. From the results achieved, we can conclude that C. aethiops is an excellent model to obtain and isolate mononuclear cells from bone marrow, fostering several studies in the field of cell therapy.

The European Union and Assembling Biofuel Development – topological investigations concerning the associations between law, policy and space

Biofuels for transport are a renewable source of energy that were once heralded as a solution to multiple problems associated with poor urban air quality, the overproduction of agricultural commodities, the energy security of the European Union (EU) and climate change. It was only after the Union had implemented an incentivizing framework of legal and political instruments for the production, trade and consumption of biofuels that the problems of weakening food security, environmental degradation and increasing greenhouse gases through land-use changes began to unfold. In other words, the difference between political aims for why biofuels are promoted and their consequences has grown – which is also recognized by the EU policy-makers. Therefore, the global networks of producing, trading and consuming biofuels may face a complete restructure if the European Commission accomplishes its pursuit to sideline crop-based biofuels after 2020. My aim with this dissertation is not only to trace the manifold evolutions of the instruments used by the Union to govern biofuels but also to reveal how this evolution has influenced the dynamics of biofuel development. Therefore, I study the ways the EU’s legal and political instruments of steering biofuels are coconstitutive with the globalized spaces of biofuel development. My analytical strategy can be outlined through three concepts. I use the term ‘assemblage’ to approach the operations of the loose entity of actors and non-human elements that are the constituents of multi-scalar and -sectorial biofuel development. ‘Topology’ refers to the spatiality of this European biofuel assemblage and its parts whose evolving relations are treated as the active constituents of space, instead of simply being located in space. I apply the concept of ‘nomosphere’ to characterize the framework of policies, laws and other instruments that the EU applies and construes while attempting to govern biofuels. Even though both the materials and methods vary in the independent articles, these three concepts characterize my analytical strategy that allows me to study law, policy and space associated with each other. The results of my examinations underscore the importance of the instruments of governance of the EU constituting and stabilizing the spaces of producing and, on the other hand, how topological ruptures in biofuel development have enforced the need to reform policies. This analysis maps the vast scope of actors that are influenced by the mechanism of EU biofuel governance and, what is more, shows how they are actively engaging in the Union’s institutional policy formulation. By examining the consequences of fast biofuel development that are spatially dislocated from the established spaces of producing, trading and consuming biofuels such as indirect land use changes, I unfold the processes not tackled by the instruments of the EU. Indeed, it is these spatially dislocated processes that have pushed the Commission construing a new type of governing biofuels: transferring the instruments of climate change mitigation to land-use policies. Although efficient in mitigating these dislocated consequences, these instruments have also created peculiar ontological scaffolding for governing biofuels. According to this mode of governance, the spatiality of biofuel development appears to be already determined and the agency that could dampen the negative consequences originating from land-use practices is treated as irrelevant.

Interhemispheric connections between primary visual areas: beyond the midline rule

In the last five years, a number of detailed anatomical, electrophysiological, optical imaging and simulation studies performed in a variety of non-human species have revealed that the functional organization of callosal connections between primary visual areas is more elaborate than previously thought. Callosal cell bodies and terminals are clustered in columns whose correspondence to features mapped in the visual cortex, such as orientation and ocularity, are starting to be understood. Callosal connections are not restricted to the vertical midline representation nor do they establish merely point-to-point retinotopic correspondences across the hemispheres, as traditionally believed. In addition, anatomical studies have revealed the existence of an ipsilateral component of callosal axons. The aim of this short review is to propose how these new data can be integrated into an updated scheme of the circuits responsible for assembling the primary visual field map.

Laboratory animal: biological reagent or living being?

The duties of humans toward non-human animals and their rights in society have been debated for a long time. However, a discussion on the terminology used for the identification of laboratory animals is usually not considered, although the employment of inadequate terminology may generate disastrous consequences for the animals before, during, and after the experiment. This study intends to defend the use of appropriate terminology, call attention to an unethical attitude of certain professionals when dealing with experimental animals, and also propose operational mechanisms, which allow for those distortions to be corrected.

Construction of bovine adenovirus type 3 E1 and E3, substitution plasmids and the sequencing analysis of DNA pol and pTP /

The relative ease to concentrate and purify adenoviruses, their well characterized mid-sized genome, and the ability to delete non-essential regions from their genome to accommodate foreign gene, made adenoviruses a suitable candidate for the construction of vectors. The use of adenoviral vectors in gene therapy, vaccination, and as a general vector system for expressing foreign genes have been documented for some time. In this study, the objective was to rescue a BAV3 E1 or E3 recombinant vector carrying the kanamycin resistant gene, a dominant selectable marker with useful applications in studying vectored gene expression in mammalian cells. To accomplish the objective of this study, more information about BAV3 DNA sequences was required in order to make the manipulation of the virus genome accessible. Therefore, sequencing of the BAV3 genome from 1 1 .7% to 30.8% was carried out. Analysis of the determined sequences revealed the primary structure of important viral gene products coded by E2 including BAV3 DNA pol and precursor to terminal protein. Comparative analysis of these proteins with their counterparts from human and non human adenoviruses revealed important insights as to the evolutionary lineage of BAV3. In order to insert the kanamycin resistance gene in either E1 or E3, it was necessary to delete BAV3 sequences to accommodate the foreign gene so as not to exceed the limit of the packaging capacity of the virus. To construct a recombinant BAV3 in which a foreign gene was inserted in the deleted E1 region, an E1 shuttle vector was constructed. This involved the deletion from the viral sequences a region between 1.3% to 9% and inserting the kanamycin resistance gene to replace the deletion. The E1 shuttle vector contained the left (0%- 53.9%) segment of the genome and was expected to generate BAV3 recombinants that can be grown and propagated in cells that can complement the missing E1 functions. To construct a similar shuttle vector for E3 deletion, DNA sequences extending from 78.9% to 82.5% (1281 bp) were deleted from within the E3 region that had been cloned into a plasmid vector. The deleted region corresponds to those that have been shown to be non-essential for viral replication in cell culture. The resulting plasmid was used to construct another recombinant plasmid with BAV3 DNA sequences extending from 37.1% to 100% and with a deletion of E3 sequences that were replaced by kanamycin resistance gene. This shuttle plasmid was used in cotransfections with digested viral DNA in an attempt to rescue a recombinant BAV3 carrying the kanamycin resistance gene to replace the deleted E3. In spite of repeated attempts of transfection, El or E3 recombinant BAV3 were not isolated. It seems that other approaches should be applied to make a final conclusion on BAV3 infectivity.

Niagara alternative food projects : networks, discourses and nature /

This thesis undertakes an exploration of the nature of alternative food projects in Niagara. A review of various theoretical approaches to the study of food and agriculture, suggests that actor-network theory offers the most useful lens through which to understand these projects. In particular, actor-network theory facilitates non-dualistic theorisations of power and scale and a commitment to the inclusion of non-humans in the 'social' sciences. The research is based on 19 in-depth interviews with actors involved in various urban and rural projects including community supported agriculture, community gardens, chefs using local seasonal food, a winery that grows organically, the good food box, a value-added small business, and organic producers. The analysis consists of four themes. The first analytical section pays special attention to the prominence of agri-tourism in Niagara, and examines the ways in which the projects in the sample interact with agri-tourist networks. In the second section the discussion focuses on the discourses and practices of resistance among Niagara alternative food actors. The participants' interviews suggest there are more discourses of resistance toward agri-tourist than toward dominant food networks. The third section questions commodity chain theorisations of alternative food projects. In particular, this section shows how the inclusion of non-human actors in an analysis confounds conceptualisations of 'short' and 'local' chains. The final analytical section assesses relations of power in Niagara alternative food projects. Three important conclusions arise from this research. First, Niagara alternative food projects cannot be conceptualised as operating at the 'local' scale. Broadening the scope of analysis to include non-human actors, it becomes apparent that these projects actually draw on a variety of extra-local actors. They are at once local and global. Second, the projects in this sample are simultaneously part of alternative, dominant and agri-tourist networks. While Niagara alternative food projects do perform many of the roles characteristic of alternative food systems, they are also involved in practices of development, business, and class distinction. Thus, alternative food networks should not be understood as separate from and in direct opposition to dominant food networks. Despite the second conclusion, this research determines that Niagara alternative food projects have made significant strides in the reworking of power. The projects represented in this thesis do engage in resistant practices and are associated with increased levels ofjustice.

Social neuroendocrinology of competition

The relationship between testosterone concentrations and aggressive behaviour in studies of people has produced very inconsistent findings. However, one consistent fmding that has emerged is that competitive and aggressive interactions potentiate testosterone release in both human and non-human species. It has been argued that socially-induced alterations in testosterone concentrations may function to influence ongoing and/or future social behaviour. Nonetheless, few studies have empirically tested this hypothesis. The current series of experiments was designed to address the extent to which competitioninduced fluctuations in testosterone concentrations were associated with ongoing and/or subsequent social behaviour. In Study 1, men (n = 38) provided saliva samples prior to, and at the conclusion of, the Point Subtraction Aggression Paradigm (PSAP). Although baseline testosterone concentrations were not related to aggressive behaviour, there was a positive correlation between change in testosterone and aggressive behaviour such that men who were most aggressive on the PSAP demonstrated the largest increase in testosterone concentrations. Furthermore, a rise in testosterone during the PSAP predicted willingness to choose a subsequent competitive task. In Study 2, men and women provided saliva samples prior to and after competing against a same-sex opponent on the Number Tracing Task (NTT). The outcome of the competition was rigged such that half of the individuals won most of the races, while the other half lost most of the races, thus experimentally creating a winner and loser in the laboratory. Following the competitive interaction, men and women played the PSAP with their same-sex partner. Results indicated that men selected the aggressive response (but not reward or protection responses), more frequently than women. For men assigned to the loss condition, an increase in testosterone concentrations in response to the NTT predicted subsequent aggressive behaviour. For men assigned to the win condition, an increase in testosterone concentrations in response to the NTT predicted subsequent aggressive behaviour, but only among those men who scored high on trait dominance. Change in testosterone and trait dominance did not predict aggressive behaviour in women. In Study 3, men provided saliva samples prior to, during, and at the end of the PSAP. They were randomly assigned to one of four experimental conditions that differed in the extent to which they were provoked and whether they received reward for behaving aggressively (i.e., stealing points). Results indicated that baseline testosterone concentrations did not correlate with aggression in any of the experimental conditions. Consistent with Study 1, there was a positive correlation between change in testosterone and aggressive behaviour among men who were provoked, but did not receive reward for aggression (i.e., reactive condition). Men who were provoked but did not receive reward for aggression enjoyed the task the most and were more likely to choose the competitive versus non-competitive task relative to men assigned to the other experimental conditions. Also, individual differences in aggressive behaviour among these men were positively correlated with the extent to which they enjoyed the task. Together, these studies indicate that testosterone dynamics within the context of competition influence subsequent competitive and aggressive behaviours in humans and that testosterone may be a marker of the intrinsically rewarding nature of costly aggressive behaviour.