Mutually Exclusive Expression of Virulence Genes by Malaria Parasites Is Regulated Independently of Antigen Production

The primary virulence determinant of Plasmodium falciparum malaria parasite–infected cells is a family of heterogeneous surface receptors collectively referred to as PfEMP1. These proteins are encoded by a large, polymorphic gene family called var. The family contains approximately 60 individual genes, which are subject to strict, mutually exclusive expression, with the single expressed var gene determining the antigenic, cytoadherent, and virulence phenotype of the infected cell. The mutually exclusive expression pattern of var genes is imperative for the parasite's ability to evade the host's immune response and is similar to the process of “allelic exclusion” described for mammalian Ig and odorant receptor genes. In mammalian systems, mutually exclusive expression is ensured by negative feedback inhibition mediated by production of a functional protein. To investigate how expression of the var gene family is regulated, we have created transgenic lines of parasites in which expression of individual var loci can be manipulated. Here we show that no such negative feedback system exists in P. falciparum and that this process is dependent solely on the transcriptional regulatory elements immediately adjacent to each gene. Transgenic parasites that are selected to express a var gene in which the PfEMP1 coding region has been replaced by a drug-selectable marker silence all other var genes in the genome, thus effectively knocking out all PfEMP1 expression and indicating that the modified gene is still recognized as a member of the var gene family. Mutually exclusive expression in P. falciparum is therefore regulated exclusively at the level of transcription, and a functional PfEMP1 protein is not necessary for viability or for proper gene regulation in cultured parasites

A regulatable transgene expression system for cultured Plasmodium falciparum parasites

<p>Abstract</p> <p>Background</p> <p>The ability to transfect and create transgenic cultured malaria parasites has transformed the study of <it>Plasmodium falciparum </it>over the last decade. With the completion of the annotated genome sequence, the process of gene discovery now routinely includes gene knockouts, over-expression and complementation analysis. However, while this technology has proven extremely valuable, significant limitations exist. In particular, <it>P. falciparum </it>DNA is often unstable and difficult to clone because of its AT-rich, repetitive nature. As a result, transgene expression constructs can be difficult to assemble due to the need to include two expression cassettes on a single plasmid, one to drive expression of the transgene of interest and a second for expression of the selectable marker. In addition, transgene expression levels are usually not regulatable, making it difficult to assess phenotypes that are sensitive to the amount of protein expressed.</p> <p>Results</p> <p>A plasmid based system for transgene expression is described that uses a single, bidirectional promoter to drive expression of both the transgene and the selectable marker, thus greatly reducing the size of the construct and enhancing stability. Further, by altering the concentration of drug used for selection, it is possible to modulate the copy number of the concatameric episomes and thereby regulate the expression level of the transgene through a range greater than 10 fold.</p> <p>Conclusion</p> <p>The transgene expression system described here should prove useful for both routine protein over-expression and complementation experiments as well as for experiments in which precisely manipulating the expression level of candidate proteins is desirable. This should provide an additional level of precision to the tools used to study the molecular biology of malaria parasites.</p

Senior Recital: Hannah Deitsch, Horn

Kemp Recital Hall November 3, 2017 Friday Evening 6:00p.m

Pregnancy-associated Malaria and the Prospects for Syndrome-specific Antimalaria Vaccines

Aided by the Plasmodium falciparum genome project, recent discoveries regarding the molecular basis of malaria pathogenesis have led to a better understanding of the interactions between host and parasite. Although vaccines that prevent infection by malaria parasites remain only hopes for the future, there are now more immediate prospects for vaccines that protect against specific disease syndromes. Here, we discuss the latest advances in the development of a vaccine that specifically targets pregnancy-associated malaria (PAM)

Creating a Millennial Generation Contextualized Church Culture

The Millennial generation, or Generation Y as some people know them, is the biggest generation in the United States of America history. As they flood schools, universities, and the job market it is easy to see that there are major differences between them and previous generations. Simultaneously, the church in America has hundreds of individual churches each year closing and tens of thousands each year declining; most churches are ceasing to grow. The inability to reach Millennials is one of the reasons for this decline. This thesis purposes to give a snapshot of the Millennial generation, overview a few of the key authors who are helping to reach them and will give churches and pastors practical components to work into their church culture to make churches and the gospel more appealing to the Millennial generation. This work is not a comprehensive solution to the problem of the church or the Millennial generation but a tool to help pastors who hope to reach the next generation

Horizontal gene transfer of epigenetic machinery and evolution of parasitism in the malaria parasite Plasmodium falciparum and other apicomplexans

Background The acquisition of complex transcriptional regulatory abilities and epigenetic machinery facilitated the transition of the ancestor of apicomplexans from a free-living organism to an obligate parasite. The ability to control sophisticated gene expression patterns enabled these ancient organisms to evolve several differentiated forms, invade multiple hosts and evade host immunity. How these abilities were acquired remains an outstanding question in protistan biology. Results In this work, we study SET domain bearing genes that are implicated in mediating immune evasion, invasion and cytoadhesion pathways of modern apicomplexans, including malaria parasites. We provide the first conclusive evidence of a horizontal gene transfer of a Histone H4 Lysine 20 (H4K20) modifier, Set8, from an animal host to the ancestor of apicomplexans. Set8 is known to contribute to the coordinated expression of genes involved in immune evasion in modern apicomplexans. We also show the likely transfer of a H3K36 methyltransferase (Ashr3 from plants), possibly derived from algal endosymbionts. These transfers appear to date to the transition from free-living organisms to parasitism and coincide with the proposed horizontal acquisition of cytoadhesion domains, the O-glycosyltransferase that modifies these domains, and the primary family of transcription factors found in apicomplexan parasites. Notably, phylogenetic support for these conclusions is robust and the genes clearly are dissimilar to SET sequences found in the closely related parasite Perkinsus marinus, and in ciliates, the nearest free-living organisms with complete genome sequences available. Conclusions Animal and plant sources of epigenetic machinery provide new insights into the evolution of parasitism in apicomplexans. Along with the horizontal transfer of cytoadhesive domains, O-linked glycosylation and key transcription factors, the acquisition of SET domain methyltransferases marks a key transitional event in the evolution to parasitism in this important protozoan lineage

Autonomous Payload Operations Onboard the International Space Station

Operating the International Space Station (ISS) involves many complex crew tended, ground operated and combined systems. Over the life of the ISS program, it has become evident that by having automated and autonomous systems on board, more can be accomplished and at the same time reduce the workload of the crew and ground operators. Engineers at the National Aeronautics and Space Administration's (NASA) Marshall Space Flight Center in Huntsville Alabama, working in collaboration with The Charles Stark Draper Laboratory have developed an autonomous software system that uses the Timeliner User Interface Language and expert logic to continuously monitor ISS payload systems, issue commands and signal ground operators as required. This paper describes the development history of the system, its concept of operation and components. The paper also discusses the testing process as well as the facilities used to develop the system. The paper concludes with a description of future enhancement plans for use on the ISS as well as potential applications to Lunar and Mars exploration systems

Malaria Epigenetics

Organisms with identical genome sequences can show substantial differences in their phenotypes owing to epigenetic changes that result in different use of their genes. Epigenetic regulation of gene expression plays a key role in the control of several fundamental processes in the biology of malaria parasites, including antigenic variation and sexual differentiation. Some of the histone modifications and chromatin-modifying enzymes that control the epigenetic states of malaria genes have been characterized, and their functions are beginning to be unraveled. The fundamental principles of epigenetic regulation of gene expression appear to be conserved between malaria parasites and model eukaryotes, but important peculiarities exist. Here, we review the current knowledge of malaria epigenetics and discuss how it can be exploited for the development of new molecular markers and new types of drugs that may contribute to malaria eradication efforts

Segmentation of Photovoltaic Module Cells in Electroluminescence Images

High resolution electroluminescence (EL) images captured in the infrared spectrum allow to visually and non-destructively inspect the quality of photovoltaic (PV) modules. Currently, however, such a visual inspection requires trained experts to discern different kinds of defects, which is time-consuming and expensive. Automated segmentation of cells is therefore a key step in automating the visual inspection workflow. In this work, we propose a robust automated segmentation method for extraction of individual solar cells from EL images of PV modules. This enables controlled studies on large amounts of data to understanding the effects of module degradation over time-a process not yet fully understood. The proposed method infers in several steps a high-level solar module representation from low-level edge features. An important step in the algorithm is to formulate the segmentation problem in terms of lens calibration by exploiting the plumbline constraint. We evaluate our method on a dataset of various solar modules types containing a total of 408 solar cells with various defects. Our method robustly solves this task with a median weighted Jaccard index of 94.47% and an $F_1$ score of 97.54%, both indicating a very high similarity between automatically segmented and ground truth solar cell masks