Robotic injection of zebrafish embryos for high-throughput screening in disease models

The increasing use of zebrafish larvae for biomedical research applications is resulting in versatile models for a variety of human diseases. These models exploit the optical transparency of zebrafish larvae and the availability of a large genetic tool box. Here we present detailed protocols for the robotic injection of zebrafish embryos at very high accuracy with a speed of up to 2000 embryos per hour. These protocols are benchmarked for several applications: (1) the injection of DNA for obtaining transgenic animals, (2) the injection of antisense morpholinos that can be used for gene knock-down, (3) the injection of microbes for studying infectious disease, and (4) the injection of human cancer cells as a model for tumor progression. We show examples of how the injected embryos can be screened at high-throughput level using fluorescence analysis. Our methods open up new avenues for the use of zebrafish larvae for large compound screens in the search for new medicines

Identifying Proteins in Zebrafish Embryos Using Spectral Libraries Generated from Dissected Adult Organs and Tissues

Proteomic

Revolutionary Memories: Historical Subjectivity and Heritage Activism in Dublin Ireland

This dissertation explores the theoretical concept of “historical subjectivity” in anthropology, which focuses on how individuals consider themselves as part of an ongoing historical narrative that extends from the past, into the present, and will be remembered in the future. This research was conducted through a case study of the ways in which activists involved with the Save Moore Street campaign in Dublin, Ireland (which revolves around saving historically significant building from being re-developed into a contemporary shopping center) invoke historical events, empathize with historical figures, and imagine how their city, and their actions, will be viewed by people in the future. Methods used are ethnographic: participant observation and in-depth interviews, and archival research (using both primary and secondary sources). The dissertation is composed of three primary standalone chapters and two sub-chapters. The Prelude sets the ethnographic present: primarily 2016 in Dublin, as the country celebrates the centenary of the 1916 Easter Rising, a seminal event in Ireland’s struggle for independence from Great Britain. Chapter 1 explores how activists use satire and invoke historical “ghosts” to appeal to the public to support their campaign, which is firmly rooted in the present state of economic uncertainty post-financial collapse. Chapter 2 introduces the idea of “historical empathy,” an idea long used by educators but less common among anthropologists. Here, I show how we in the present can empathize with historical figures through points of connection: family lineage, using similar space, and even watching their actions on film, which can meaningfully affect us and motivate our actions in the present and for the future. The Ethnographic Interlude continues this discussion with a short section on ideas of respect for historical figures in a commemorative setting. Finally, Chapter 3 focuses on how activists and developers imagine the future according to ethical and economic goals, which I argue are not entirely antithetical in their goals of urban revitalization

Ghosts of the Lizzie Borden House Tour: Hauntology, Historicity and Attention at Work

This thesis combines work in hauntology and phenomenology to explore the ways in which individuals experience the history of the Lizzie Borden House, especially on a tour of the house guided by a member of the staff. "Hauntology" is a concept developed primarily by Jacques Derrida in order to explore the lingering injustices associated with Western fears of communism (1994). However, it has been further utilized by diverse scholars, such as Avery Gordon (2008) and Byron Good (2012) to study how unspeakable traumas are engaged by literature and ethnographic subjects. My work will expand and connect these discrete lines of scholarship through the ethnographic and historical legacy of an infamous American murder scene that is now a privately owned and operated bed and breakfast

Ghosts of the Lizzie Borden House Tour: Hauntology, Historicity and Attention at Work

This thesis combines work in hauntology and phenomenology to explore the ways in which individuals experience the history of the Lizzie Borden House, especially on a tour of the house guided by a member of the staff. "Hauntology" is a concept developed primarily by Jacques Derrida in order to explore the lingering injustices associated with Western fears of communism (1994). However, it has been further utilized by diverse scholars, such as Avery Gordon (2008) and Byron Good (2012) to study how unspeakable traumas are engaged by literature and ethnographic subjects. My work will expand and connect these discrete lines of scholarship through the ethnographic and historical legacy of an infamous American murder scene that is now a privately owned and operated bed and breakfast

Current perspectives on the interleukin-1 family as targets for inflammatory disease.

Since the ?rst description of interleukin-1 (IL-1) and the genesis of the ?eld of cytokine biology, the understanding of how IL-1 and related cytokines play central orchestrat- ing roles in the in?ammatory response has been an area of intense investigation. As a consequence of these endeavours, speci?c strategies have been developed to target the function of the IL-1 family in human disease realizing signi?cant impacts for patients. While the most signi?cant advances to date have been associated with inhibition of the prototypical family members IL-1?/?, approaches to target more recently identi?ed family members such as IL-18, IL-33 and the IL-36 subfamily are now beginning to come to fruition. This review summarizes current knowledge surrounding the roles of the IL- 1 family in human disease and describes the rationale and strategies which have been developed to target these cytokines to inhibit the pathogenesis of a wide range of diseases in which in?ammation plays a centrally important role

Simvastatin Suppresses Interleukin Iβ Release in Human Peripheral Blood Mononuclear Cells Stimulated With Cholesterol Crystals

Statins are mainstream therapy in the treatment and prevention of cardiovascular disease through inhibitory effects on cholesterol synthesis. However, statins’ beneficial effects in cardiovascular disease may also be attributable to their role as anti-inflammatory mediators. Here, we investigated the effects of simvastatin treatment on expression levels of interleukin (IL) 1β in both patient with hyperlipidemia and healthy human peripheral blood mononuclear cells (PBMCs) using cholesterol crystals (CC), a cardiovascular pathogenic stimulus for activation of the NOD-like receptor pyrin domain–containing protein 3 (NLRP3) inflammasome. Cholesterol crystal-induced NLRP3 inflammasome activation was used to trigger maturation and release of IL-1β in PBMCs. Specifically, isolated PBMCs from patients with hyperlipidemia at baseline and following 8 weeks of in vivo treatment with simvastatin (10-20 mg) daily were stimulated with lipopolysaccharide (LPS; 100 ng/mL) for 3 hours to induce proIL-Iβ expression followed by CC (2 mg/mL) stimulation for further 18 hours to activate the NLRP3 inflammasome complex to induce maturation/activation of IL-1β. Peripheral blood mononuclear cells were also isolated from healthy donors and stimulated in vitro with simvastatin (50, 25, 5, and 2 µmol/L) prior to stimulation with LPS and CC as described above. The effects of simvastatin treatment on levels of IL-1β expression were determined by enzyme-linked immunosorbent assay and western blot. Both in vitro and in vivo treatments with simvastatin led to a significant reduction in the levels of expression of IL-1β in response to stimulation with CC. Simvastatin inhibits the expression and activation of IL-1β induced by CC in PBMCs, which may contribute to its protective role in patients with cardiovascular disease.</jats:p