Synthesis and characterization of a masked N-heterocyclic phosphinidene (NHP)

This thesis summarizes the synthesis and characterization of an unprecedented heterocyclobutane derivative. To assess the biradical character, quantum mechanical computations were carried out. The second part elaborates on the reactivity (LEWIS acids/bases, an isonitrile, carbon monoxide, chalcogens). Notably, the addition of alkenes/alkynes to form phosphiranes/phosphirenes highlights the phosphinidene-type reactivity. In the last part, heavier congeners (As, Sb, Bi) of chlorinated precursors and theoretical investigations of their corresponding reduced heterocycles is discussed.Die Arbeit behandelt die Synthese und Charakterisierung eines neuartigen Heterocyclobutanderivats. Hierbei wurden quantenmechanische Berechnungen durchgeführt, um den Biradikalcharakter abzuschätzen. Im zweiten Teil wurde die Reaktivität untersucht (LEWIS-Säuren & –Basen, ein Isonitril, CO, Chalcogene). Die Addition von Alkenen/Alkinen unter Ausbildung von Phosphiranen/Phosphirenen verdeutlicht die Phosphiniden-Reaktivität. Der letzte Teil behandelt die Synthese schwerer Homologa (As, Sb, Bi) chlorierter Vorläuferverbindungen und theoretische Untersuchungen zu cyclisierten reduzierten Spezies

SEA CUCUMBER (Apostichopus californicus) CULTURE AND ENHANCEMENT OPPORTUNITIES.

Demand for giant red sea cucumbers (Apostichopus californicus) continues in Asian markets while wild harvest has declined on the west coast of North America. Multiple trials are in place in Washington state to culture this species on upland and floating structures that support other species. Co-culture of sea cucumbers has proved to be effective with mussels, black cod, sea weed and oysters. In addition, there are food resources from aquaculture that the wild population currently exploits but conceivably could utilize more efficiently. Efforts to further this utilization are in progress

Sizing and morphology analysis of milled olanzapine, paroxetine and gabapentin particles using hot stage microscopy and laser diffractometry

Poster presented at the 2nd International Congress of CiiEM: Translational Research and Innovation in Human and Health Sciences. Egas Moniz, Caparica, Portugal, 11-13 June 2017N/

Mucosal vaccination with a live recombinant rhinovirus followed by intradermal DNA administration elicits potent and protective HIV-specific immune responses

Published: 17 November 2016Mucosal immunity is deemed crucial to control sexual transmission of human immunodeficiency virus (HIV). Herein we report the efficacy of a mucosal HIV vaccine strategy comprising intranasal (IN) vaccination with a cocktail of live recombinant human rhinoviruses (HRVs) encoding overlapping fragments of HIV Gag and full length Tat (rHRV-Gag/Tat) followed by intradermal (ID) vaccination with DNA vaccines encoding HIV Gag and Tat (pVAX-Gag-Tat). This heterologous prime-boost strategy will be referred to hereafter as rHRV-DNA. As a control, IN vaccination with wild type (wt)-HRV-A1 followed by a single ID dose of pVAX (wt-HRV-A1/pVAX vaccination) was included. rHRV-DNA vaccination elicited superior multi-functional CD8(+)T cell responses in lymphocytes harvested from mesenteric lymph nodes and spleens, and higher titres of Tat-specific antibodies in blood and vaginal lavages, and reduced the viral load more effectively after challenge with EcoHIV, a murine HIV challenge model, in peritoneal macrophages, splenocytes and blood compared compared with wt-HRV-A1/pVAX vaccination or administration of 3 ID doses of pVAX-Gag-Tat (3X pVAX-Gag-Tat vaccination). These data provide the first evidence that a rHRV-DNA vaccination regimen can induce HIV-specific immune responses in the gut, vaginal mucosa and systemically, and supports further testing of this regimen in the development of an effective mucosally-targeted HIV-1 vaccine.Khamis Tomusange, Danushka Wijesundara, Jason Gummow, Steve Wesselingh, Andreas Suhrbier, Eric J. Gowans, Branka Grubor-Bau

Priming of CD8+ T cell responses following immunization with heat-killed Plasmodium sporozoites.

Protective immune responses against malaria are induced by immunization with radiation-attenuated Plasmodium sporozoites. In contrast, non-viable, heat-killed sporozoites do not induce protection, emphasizing the requirement for live parasites to achieve effective immune responses. Using an experimental system with CD8+ T cells from T cell receptor-transgenic mice, we analyzed the primary CD8+ T cell responses elicited by heat-killed inactivated sporozoites. We found that the numbers of specific CD8+ T cells induced were much lower compared to when immunizing with attenuated sporozoites; however, the kinetics of activation and the phenotype of these T cells were similar in both groups. Despite their low frequency after priming, high numbers of specific CD8+ T cells were observed after boosting with a recombinant vaccinia virus. Upon induction of the recall response, the same level of protection was observed when either heat-killed or attenuated sporozoites were used for priming. We propose that live parasites are not critical for the induction of memory T cell populations against the malaria liver stages

Shellfish at Work: Nutrient Bioextraction Demonstration in South Puget Sound

This project explored the application of extractive aquaculture technologies using shellfish for nutrient mitigation in urban nearshore environments of Tacoma and Olympia, WA. Nutrient removal by shellfish, which are then harvested and removed from the system, has the potential to help address environmental issues such as excess inputs of nutrients (eutrophication), low dissolved oxygen, and reduced light availability. The research focus was to: 1) implement demonstration aquaculture systems; 2) determine nutrient removal services of mussels grown; 3) assess the feasibility of producing soil compost from the demonstration systems; and 4) increase local stakeholder awareness of and engagement in urban water quality mitigation needs and strategies. In Budd Inlet over 225 nylon straps were placed under 3 existing dock structures to provide a suitable substrate for mussel recruits. By summer’s end, over 3,600 kg (over 8,000 lbs) of adult mussels were filter-feeding on these straps. These mussels were harvested, chipped, and delivered to The Evergreen State College’s Organic Farm, WSU-Puyallup Research and Extension Center, and Cedar Creek Correctional Center for ongoing compost trials. The mussels removed an estimated 360 kg (800 lbs) of nitrogen (wet weight) from lower Budd Inlet over the course of 4 months. In addition, to the bioextraction trials, outreach efforts focused on improving public understanding of local water quality issues, ecosystem services of shellfish, and controlling upland sources of nutrients that flow into Puget Sound

A poética da fome na arte Guarani.

Drawings by Guarani Mbyá children at the turning of the millenium suggest that the Land-without-Evils may be a worldly reality. The barren and plagueful soil of the Indigenous Land Itaóca, at the southern coast of São Paulo, is transformed by young artists into a rich territory, full of plantation and fit for hunting. The proximity of the big garbage dump of Mongaguá, banana farms and the “Equality Cemetery”, where the Guarani search for food, work as slaves and prematurely bury their deads, is kept out of the illustrations. Sick and weakened children from the most numerous Indian group of Brazil (30.000) materialize in the illustrations in xandaro Guarani - warriors whose bodies encapsulated the essence of the mytical paradise, the ¿mortality. The aesthetic quality of the representations of the Guarani social life comes from the dimensions of an erring world which the children try to recreate and express through art. Whereas adults and elder people belive that famine and scarcity are necessary conditions for the passage to the Land-without-Evils, the newer generation proposes concrete changes to social order, including the acceptance of the comfort of the sedentary agriculture. If the sacred word, vital to the Guarani person, did not transmit, up to now, the dramatic effects of poverty and violence to the unsensitive Brazilian society, the young hope that visual signs of a poetic and idealized world may educate the people about their present aspirations.Desenhos produzidos por crianças Guarani Mbyá na virada do milênio sugerem que a Terra-sem-Males pode ser uma realidade mundana. O solo infértil e cheio de pragas da Terra Indígena Itaóca no litoral sul de São Paulo é transformado por jovens artistas em farto território, repleto de planta­ ções e caça. A proximidade do lixão de Mongaguá, fazendas de banana e Cemitério da Igualdade, onde os Guarani buscam restos de alimento, trabalham como escravos e enterram seus mortos prematuramente, é mantida fora das ilustrações. Crianças doentes e enfraquecidas do grupo indígena mais numeroso do Brasil (30.000) materializam-se nas ilustrações em xondaro Guarani - guerreiros cujos corpos captaram a essência do paraíso mítico, a imortalidade. A qualidade estética das representações da vida social Guarani advém das dimensões de um mundo errante que as crianças tentam recriar e expressar por meio da arte. Enquanto adultos e idosos acreditam que fome e escassez são condições necessárias para a passagem à Terra-sem-Males, a geração mais nova propõe mudanças concretas à ordem social, incluindo a aceitação do conforto da agricultura sedentária. Se a palavra sagrada, vital à pessoa Guarani, não transmitiu até hoje à insensível sociedade brasileira os efeitos dramáticos da pobreza e violência, os jovens esperam que sinais visuais de um mundo poético e idealizado possam educar o público sobre suas presentes aspirações

Kunjin virus replicons: an RNA-based, non-cytopathic viral vector system for protein production, vaccine and gene therapy applications

The application of viral vectors for gene expression and delivery is rapidly evolving, with several entering clinical trials. However, a number of issues, including safety, gene expression levels, cell selectivity and antivector immunity, are driving the search for new vector systems. A number of replicon-based vectors derived from positive-strand RNA viruses have recently been developed, and this paper reviews the current knowledge on the first flavivirus replicon system, which is based on the Australian flavivirus Kunjin NUN). Like most replicon systems, KUN replicons can be delivered as DNA, RNA or virus-like particles, they replicate their RNA in the cytoplasm and direct prolonged high-level gene expression. However, unlike most alphavirus replicon systems, KUN replicons are non-cytopathic, with transfected cells able to divide, allowing the establishment of cell lines stably expressing replicon RNA and heterologous genes. As vaccine vectors KUN replicons can induce potent, long-lived, protective, immunogen-specific CD8(+) T cell immunity, a feature potentially related to extended production of antigen and double-stranded RNA-induced 'danger signals'. The identification of KUN replicon mutants that induce increased levels of IFN-alpha/beta has also spawned investigation of KUN replicons for use in cancer gene therapy. The unique characteristics of KUN replicons may thus make them suitable for specific protein production, vaccine and gene therapy applications

Heat shock protein 10 inhibits lipopolysaccharide-induced inflammatory mediator production

Heat shock protein 10 (Hsp10) and heat shock protein 160 (Hsp60) were originally described as essential mitochondrial proteins involved in protein folding. How,ever, both proteins have also been shown to have a number of extracellular immunomodulatory activities. Here we show that purified recombinant human Hsp10 incubated with cells in vitro reduced lipopolysaccharide (LPS)-induced nuclear factor-kappaB activation and secretion of several inflammatory mediators from RAW264.7 cells, murine macrophages, and human peripheral blood mononuclear cells. Induction of tolerance by contaminating LPS was formally excluded as being responsible for Hsp10 activity. Treatment of mice with Hsp10 before,endotoxin challenge resulted in the reduction of serum tumor necrosis factor-a and RANTES (regulated upon activation, normal T cell expressed and secreted) levels and an elevation of serum interleukin-10 levels. Hsp10 treatment also delayed mortality in a murine graft-ver-sus-host disease model, where gut-derived LPS contributes to pathology. We were unable to confirm previous reports that Hsp10 has tumor growth factor properties and suggest that Hsp10 exerts anti-inflammatory activity by inhibiting Toll-like receptor signaling possibly by interacting with extracellular Hsp60