Determining the Individual Roles of Telomere-Associated Proteins in Telomere Regulation in Arabidopsis

Telomeres consist of repeating sequences of nucleotides at the end of linear chromosomes and associated proteins that together serve to protect chromosome ends from degradation or fusion. Misregulation of telomeres leads to issues such as replication stress, DNA double-strand breaks, and other biological consequences that affect normal DNA replication and organismal viability. Telomerase—an essential enzyme for telomere function and maintenance—is responsible for telomere replication and the maintenance of telomere length. In Arabidopsis, studies have led to the discovery of a highly conserved and essential telomere protein complex CTC1/STN1/TEN1 (CST), which caps and protects the end of the chromosomes. The CTC1 and STN1 components of CST complex in Arabidopsis interact with telomere-associated proteins POT1a and TEN1, which positively and negatively regulate telomerase activity, respectively. This study will investigate how the individual telomere-associated proteins CTC1, STN1, TEN1, and POT1a interact with each other, with the telomere, and with telomerase in vivo. Mutations in amino acids important for mediating the physical interactions between these proteins will be introduced, and their effects on telomere regulation will be tested. Analyzing the biological consequences of these mutations, and their effect on telomere integrity, will give insight to the individual roles of telomere-associated proteins in vivo and advance understanding of how telomeres provide genome stability. As events caused by telomere dysfunction occur in a myriad of diseases related to defects in DNA and cell replication, the study of telomeres, specifically the CST complex, may provide insight into stem cell-related diseases and cancer

Isolation of Cyanide Hydratase Mutants from Gloeocerospora Sorghi at alkaline pH

Cyanide is both a useful and dangerous chemical compound that serves as a crucial component in multiple industrial processes, including metal mining. The leaching process that utilizes cyanide ions to help separate target metals and increase mining yield is an industrial standard for chemical leaching. However, this method of ore extraction results in toxic cyanide waste that requires dangerous, costly, and potentially environmentally damaging remediation systems to degrade. As cyanide is a naturally occurring substance, several organisms contain enzymes capable of oxidizing cyanide into less toxic compounds. Despite the effectiveness of these proteins, they lack stability and functionality at the alkaline pH levels industrial cyanide is stored at. This project attempts to optimize the screening and mutagenesis methods in hopes of a isolating an alkaline tolerant mutant of cyanide hydratase, an enzyme originally found in the fungus Gloeocercospora sorghi. This approach incorporates random mutagenesis of the target fungal gene using error-prone polymerase chain reaction and an in vivo picric acid assay that tests the activity of the mutant enzymes at target conditions. Experimentation was used to determine the ideal conditions for a screening method by testing the activity of the wild-type positive control at different reaction conditions. The final, optimized screening conditions for the high throughput assay combined a 50 μL aliquot of cell culture grown overnight in a 96 well plate with a 50 μL of 0.1 M CAPS buffered to pH 10.5. As screening continues, these conditions can be used to identify a viable, alkaline tolerant mutant. If such a mutant is identified, the molecule would be a strong bioremediation candidate for the metal mining industry and could lead to more efficient and environmentally friendly degradation of cyanide waste

Brutality

I. There is something I want to say about brutality. It comes out the lips of the hateful mother, the beautiful mother

On stability of free laminar boundary layer between parallel streams

An analysis and calculations on the stability of the free laminar boundary layer between parallel streams were made for an incompressible fluid using the Tollmien-Schlichting theory of small disturbances. Because the boundary conditions are at infinity, two solutions of the Orr-Sommerfeld stability equations need not be considered, and the remaining two solutions are exponential in character at the infinite boundaries. The calculations show that the flow is unstable except for very low Reynolds numbers

Negotiating Contested Identities: Palestinian American Identity in Poems by Lisa Suhair Majaj, Naomi Shihab Nye, and Susan Abulhawa

Denne masteroppgaven tar for seg poetiske fremstillinger av sammensatte identiteter i verkene til tre Palestinsk Amerikanske samtidsdiktere som, direkte eller indirekte, har det komplekse forholdet mellom USA og Palestina som et bakteppe. Til tross for at både diplomatiske og geopolitiske perspektiver står sentralt i diskusjonen av en slik omstridt og kompleks identitet, vil oppgaven ikke gå i dybden på den aktuelle politiske situasjonen; her vil derimot den litterære fremstillingen stå i sentrum. De fire utvalgte diktene som danner grunnlaget for analysen er hentet fra Lisa Suhair Majajs Geographies of Light (2009), Naomi Shihab Nyes 19 Varieties of Gazelle (2002) og Susan Abulhawas My Voice Sought the Wind (2013). Et mangfold av perspektiver trer fram når oppgaven tar for seg hvordan de tre kvinnene bruker diktning til å illustrere utfordringen i det å skulle balansere en iboende dobbelthet i sin egen eksistens. Av den grunn blir denne masteroppgaven et sentralt bidrag til et voksende litterært felt av Arabisk Amerikansk litteratur som, så vidt meg bekjent, har en overvekt av oversiktsverk snarere enn studier basert på nærlesning.Engelsk mastergradsoppgaveMAHF-LÆFRMPENGLMAHF-ENGZMPHFFAKENG35

Mathematics Departmental Strategic Plan: The Case of Mufulira College of Education

The purpose of this study was to find out why the mathematics department at Mufulira College of Education (MUCE) does not use strategic planning in its planning. A case study was undertaken at MUCE, a public college on the Copperbelt province of Zambia. The research was conducted on twenty two (22) mathematics students, five (5) mathematics lecturers and the client organization (Principal). In total, the sample size was 28 as already tabulated above. Questionnaires were used to collect data for this study. The research design used for this study was a mixed survey method. Data collected were analyzed quantitatively using the Likert’s 5 Scale model. The results obtained reviewed that mathematics lecturers had little or limited knowledge of strategic planning. The findings of the study indicated that, the department has had no written strategic plan since it was upgraded as a secondary school teacher training college in 2008. Consequently, the current planning system is not near organized strategic planning. To this effect, the department needs to develop a strategic plan for improved performance

Caring to make a difference with vulnerable women: the impact of targeted support on birth-related outcomes and experiences

We report findings from a mixed-methods study to compare birth outcomes and experiences between vulnerable women who had or had not received targeted support from a specialist midwifery team and/or a third-sector organisation. Sociodemographic and birth-related outcome data from a 12-month birth cohort were used to explore differences between a) vulnerable and non-vulnerable women; and b) vulnerable women who had and those who had not received targeted support. Seventeen women who had or had not received targeted support were interviewed. Vulnerable mothers were significantly more likely to be younger, from a minority ethnic group, access care later in pregnancy and have a baby born earlier and at a lower birth weight. A higher percentage of women who received targeted support had a spontaneous birth, a vaginal presentation at birth and were less likely to use anaesthesia during labour when compared with unsupported vulnerable women. Targeted support was associated with reassurance, increased parental confidence and wellbeing. Key practice and policy-based implications are considered

Deciphering the heterogeneity of the Lyve1+ perivascular macrophages in the mouse brain

Perivascular macrophages (pvMs) are associated with cerebral vasculature and mediate brain drainage and immune regulation. Here, using reporter mouse models, whole brain and section immunofluorescence, flow cytometry, and single cell RNA sequencing, besides the Lyve1+F4/80+CD206+CX3CR1+ pvMs, we identify a CX3CR1– pvM population that shares phagocytic functions and location. Furthermore, the brain parenchyma vasculature mostly hosts Lyve1+MHCII– pvMs with low to intermediate CD45 expression. Using the double Cx3cr1GFP x Cx3cr1-Cre;RosatdT reporter mice for finer mapping of the lineages, we establish that CD45lowCX3CR1– pvMs are derived from CX3CR1+ precursors and require PU.1 during their ontogeny. In parallel, results from the Cxcr4-CreErt2;Rosa26tdT lineage tracing model support a bone marrow-independent replenishment of all Lyve1+ pvMs in the adult mouse brain. Lastly, flow cytometry and 3D immunofluorescence analysis uncover increased percentage of pvMs following photothrombotic induced stroke. Our results thus show that the parenchymal pvM population is more heterogenous than previously described, and includes a CD45low and CX3CR1– pvM population

Investigation of translation elongation, termination and recycling by bulk biochemical and single molecule methods

The process of translation of mRNAs into functional proteins by the ribosome is crucial to cell viability. Problems encountered by the ribosome during any of the phases of this process can have major deleterious effects. Although, our understanding of the steps of the translation have grown immensely over the years, specific questions regarding the complex dynamics of these processes have yet to be fully understood. In particular, recent studies have elucidated pauses during elongation that are caused by specific pairs of codons. Through the use of in vitro biochemical experiments in tandem with and in vivo ribosome profiling and cryo-EM structures to investigate these inhibitory codon pairs, we were able to demonstrate that this type of elongation stalling is mainly caused by slow decoding in the ribosomal A site. The mechanistic details of how the ribosome is rescued from stalls similar to these during elongation, as well as the similar processes of termination and recycling, also remain to be fully understood. Through my development of a single molecule fluorescence microscopy assay to investigate ribosome rescue, termination and recycling these dynamics can be further explored. Taken together, I have furthered our understanding of a particular type of elongation pause using an in vitro biochemical system and have expanded that system for use at the single molecule level to investigate the dynamic processes of translation further