Utilizing Single-Molecule Fret Methods to Study Conformational Changes in Trim5α

Single-molecule FRET (smFRET) is a method by which dynamic conformational changes can be monitored in a protein microscopically and in real time. smFRET relies on the creation of FRET (Förster Resonance Energy Transfer) between small molecule fluorophores conjugated to the biomolecules of interest. FRET efficiency allows calculation of interfluorophore distances. Changes in FRET efficiency represent changes in protein conformation which can inform further structural and molecular studies of the protein of interest. For example, in the Campbell Lab, we study the protein TRIM5α, an antiretroviral cellular protein which can cause premature dissociation of the HIV capsid core by an unknown mechanism. We can conjugate small fluorophores to specific sites chosen on TRIM5α or isolated TRIM5α domains and use smFRET to observe conformational changes in TRIM5α that may correlate to disruption of the viral core. Fluorophore conjugation can be done several ways, but we will focus on the use of cysteines to conjugate maleimide linked fluorophores to our protein of interest. For my thesis, I propose to adapt and optomise a protocol for smFRET usable by the Campbell Lab. To do this, I will first purify, label, and characterize peptides derived from the CC and L2 domains of TRIM5α that retain their secondary structure. I will then optimize methods for smFRET including instrument calibration, setting up the ability to track multiple fluorophores simultaneously, and analysis of smFRET data both to correctly align data and to render final data analyses

The Role Of Leadership Preparation In Teacher Leaders' Formation Of Critical Consciousness And Enactment Of Critical Teacher Leadership

Thesis (Ph.D.) - Indiana University, School of Education, 2008Research has shown that teachers feel alienated by conventional constructions of school leadership, and therefore, seek leadership opportunities that are collaborative; teachers are also more likely to become teacher leaders in environments where they feel appreciated and believe in the direction of current leadership structures and philosophy at their schools (Lambert, 2003). Conversely, it is possible that teachers who feel unsatisfied may seek teacher leadership opportunities in order to alter the current direction of the school or even the education system as a whole. Such teacher leaders are considered critical and work for a more equitable and just educational system. But whether critical teacher leaders exist and the nature of such leadership is yet to be determined. For these reasons, it is important to further study the critical nature of teacher leadership. Data was gathered for this study using critical qualitative methodology, a theoretical approach grounded in critical theory (Carspecken, 1996). Three data sources were collected for this study. First, participants were observed in four of the educational leadership courses. Second, assignments from three of the participants' educational leadership courses were collected as a check to determine if their written work reflected the ideas they expressed during class. Third, four key informants were interviewed three times to discover how class meetings and assignments affected individual teachers. Findings indicated that the formation of critical consciousness rests in course content, and professor and class member interactions. Not only do teacher leaders need to be exposed to readings that probe them to think critically, but they also need safe and collaborative opportunities to challenge their own and each other's thinking within courses. The preparatory experiences of teacher leaders affect their conception of teacher leadership as well as how they enact teacher leadership in their schools

Protein Malnutrition and the Febrile Response in the Fischer Rat

We assessed the effect of protein deprivation on the ability of peritoneal macrophages from Fischer rats to produce interieukin‐1 (IL‐1) after in vitro stimulation. Pyrogenic activity of supernatants was measured by an in vivo febrile response assay. Control rats were given a 23% casein diet and protein‐malnourished rats were given an 8% casein diet for 4 weeks. IL‐1‐containing supernatants prepared from peritoneal macrophages were injected into assay rats, whose temperatures were measured for 6 hours (δT6). Rats injected with IL‐1‐containing supernatants derived from peritoneal macrophage cultures of protein‐deprived rats had significantly less fever (δT6 = 0.20 ± 0.09°) than rats injected with IL‐1 containing supernatants derived from peritoneal macrophage cultures of control rats (δT6 = 0.56 ± 0.09°), P < .01. Protein malnutrition leads to diminished pyrogenicity of macrophage culture supernatants and may be at least partly responsible for the decreased febrile response seen in the malnourished animals.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/142038/1/jlb0036.pd

Early occipital sensitivity to syntactic category is based on form typicality

Syntactic factors can rapidly affect behavioral and neural responses during language processing; however, the mechanisms that allow this rapid extraction of syntactically relevant information remain poorly understood. We addressed this issue using magnetoencephalography and found that an unexpected word category (e.g., “The recently princess . . . ”) elicits enhanced activity in visual cortex as early as 120 ms after exposure, and that this activity occurs as a function of the compatibility of a word’s form with the form properties associated with a predicted word category. Because no sensitivity to linguistic factors has been previously reported for words in isolation at this stage of visual analysis, we propose that predictions about upcoming syntactic categories are translated into form-based estimates, which are made available to sensory cortices. This finding may be a key component to elucidating the mechanisms that allow the extreme rapidity and efficiency of language comprehension

Bilinear Optimal Control of the Velocity Term in a Kirchhoff Plate Equation

AbstractWe consider a bilinear optimal control problem where the state equation is a Kirchhoff plate equation. The control acts as a multiplier of the velocity term. We prove the existence of an optimal control in a class h∈UM={h∈L∞(0,T); −M≤h(t)≤M} and uniqueness of this optimal control for T sufficiently small

Fever of Unknown Origin in the Elderly: Lymphoma Presenting as Vertebral Compression Fractures

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/111209/1/j.1532-5415.1994.tb06080.x.pd

Susceptibility of Yeast-Like Fungi to a New Antifungal Agent, LY 121019

LY 121019, a new antifungal antibiotic agent, was tested for activity against 200 clinical isolates of Candida and other yeast-like fungi. LY 121019 had its greatest inhibitory effect on C. albicans , and C. tropicalis. C. glabrata and most other Candida species were not as sensitive. Cryptococcus and other yeast-like fungi, with the exception of a few strains, were not susceptible to LY 121019. Zusammenfassung :  LY 121019, ein neues Antimyzetikum, wurde auf seine AktivitÄt gegen 200 klinische Isolate von Candida und anderen, hefeÄhnlichen Pilzen untersucht. LY 121019 hat seine grÖßte Hemmwirkung gegen C. albicans und C. tropicalis. C. glabrata und die meisten anderen Candida -Arten waren nicht so empfindlich. Cryptococcus und andere, hefeÄhnliche Pilze waren, mit Ausnahme weniger StÄmme, nicht fÜr LY 121019 empfindlich.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/73892/1/j.1439-0507.1988.tb04426.x.pd