Doctor of Philosophy

dissertationDevelopment of the nervous system involves establishment of precise long distance connections between distinct single cells called neurons. During development, long axons extend from the cell body and grow, following a very specific pathway through a complex environment to their target. The growth cone is a dynamic structure with finger-like filopodia that sense guidance cues in the surrounding environment through receptors. External guidance cues can be attractive or repulsive, and growth cone turning in response to a guidance cue is driven by actin dynamics, with increased polymerization during attractive turning and increased disassembly during repulsive turning. Therefore, regulators of actin dynamics such as actin-binding proteins are the targets of signaling in the growth cone initiated by external guidance cues. The growth cone changes its behavior very rapidly in response to guidance cues, even as it becomes further and further from the cell body. Growth cones have the ability to act autonomously to guidance cues, in order to continue to react quickly without delay. Local mRNA translation in growth cones has an important role in growth cone behavior, giving the ability to respond to external guidance cues without communication with the cell body. Local translation is regulated by RNA-binding proteins and directly influences actin dynamics that are important for growth cone behavior. While in vitro studies have revealed a wealth of knowledge about the mechanisms involved with local translation during axon guidance, the requirement for the function of RNA-binding proteins in vivo has not been tested extensively. In this dissertation, an in vivo local translation timelapse assay, performed in the zebrafish retinotectal system, demonstrates that the zebrafish β- actin3'UTR is sufficient to target Kaede expression to RGC growth cones. Also, Igf2bp1 is shown to be the zebrafish ZBP1 ortholog, and a bipartite "zipcode element," required for interaction with ZBP1, is identified in the β-actin3'UTR. I also present the first evidence to date that Igf2bp1/ZBP1 function is required for axon guidance, and furthermore provide data that suggest that Igf2bp1 function may be required for axon specification or elongation, contrary to the common belief that ZBP1 function is required only for axon branching and arbor formation, and that local β-actin translation is only required for growth cone turning during axon guidance

Can We Really Prevent Suicide?

Every year, suicide is among the top 20 leading causes of death globally for all ages. Unfortunately, suicide is difficult to prevent, in large part because the prevalence of risk factors is high among the general population. In this review, clinical and psychological risk factors are examined and methods for suicide prevention are discussed. Prevention strategies found to be effective in suicide prevention include means restriction, responsible media coverage, and general public education, as well identification methods such as screening, gatekeeper training, and primary care physician education. Although the treatment for preventing suicide is difficult, follow-up that includes pharmacotherapy, psychotherapy, or both may be useful. However, prevention methods cannot be restricted to the individual. Community, social, and policy interventions will also be essentia

Using Automated Health Plan Data to Assess Infection Risk from Coronary Artery Bypass Surgery

We determined if infection indicators were sufficiently consistent across health plans to allow comparison of hospitals’ risks of infection after coronary artery bypass surgery. Three managed care organizations accounted for 90% of managed care in eastern Massachusetts, from October 1996 through March 1999. We searched automated inpatient and outpatient claims and outpatient pharmacy dispensing files for indicator codes suggestive of postoperative surgical site infection. We reviewed full text medical records of patients with indicator codes to confirm infection status. We compared the hospital-specific proportions of cases with an indicator code, adjusting for health plan, age, sex, and chronic disease score. A total of 536 (27%) of 1,953 patients had infection indicators. Infection was confirmed in 79 (53%) of 149 reviewed records with adequate documentation. The proportion of patients with an indicator of infection varied significantly (p<0.001) between hospitals (19% to 36%) and health plans (22% to 33%). The difference between hospitals persisted after adjustment for health plan and patients’ age and sex. Similar relationships were observed when postoperative antibiotic information was ignored. Automated claims and pharmacy data from different health plans can be used together to allow inexpensive, routine monitoring of indicators of postoperative infection, with the goal of identifying institutions that can be further evaluated to determine if risks for infection can be reduced

Is group cognitive behaviour therapy for postnatal depression evidence-based practice? A systematic review

Background: There is evidence that psychological therapies including cognitive behaviour therapy (CBT) may be effective in reducing postnatal depression (PND) when offered to individuals. In clinical practice, this is also implemented in a group therapy format, which, although not recommended in guidelines, is seen as a cost-effective alternative. To consider the extent to which group methods can be seen as evidence-based, we systematically review and synthesise the evidence for the efficacy of group CBT compared to currently used packages of care for women with PND, and we discuss further factors which may contribute to clinician confidence in implementing an intervention. Methods: Seventeen electronic databases were searched. All full papers were read by two reviewers and a third reviewer was consulted in the event of a disagreement on inclusion. Selected studies were quality assessed, using the Cochrane Risk of Bias Tool, were data extracted by two reviewers using a standardised data extraction form and statistically synthesised where appropriate using the fixed-effect inverse-variance method. Results: Seven studies met the inclusion criteria. Meta-analyses showed group CBT to be effective in reducing depression compared to routine primary care, usual care or waiting list groups. A pooled effect size of d = 0.57 (95% CI 0.34 to 0.80, p < 0.001) was observed at 10–13 weeks post-randomisation, reducing to d = 0.28 (95% CI 0.03 to 0.53, p = 0.025) at 6 months. The non-randomised comparisons against waiting list controls at 10–13 weeks was associated with a larger effect size of d = 0.94 (95% CI 0.42 to 1.47, p < 0.001). However due to the limitations of the available data, such as ill-specified definitions of the CBT component of the group programmes, these results should be interpreted with caution. Conclusions: Although the evidence available is limited, group CBT was shown to be effective. We argue, therefore, that there is sufficient evidence to implement group CBT, conditional upon routinely collected outcomes being benchmarked against those obtained in trials of individual CBT, and with other important factors such as patient preference, clinical experience, and information from the local context taken into account when making the treatment decision

Serratamolide is a hemolytic factor produced by Serratia marcescens

Serratia marcescens is a common contaminant of contact lens cases and lenses. Hemolytic factors of S. marcescens contribute to the virulence of this opportunistic bacterial pathogen. We took advantage of an observed hyper-hemolytic phenotype of crp mutants to investigate mechanisms of hemolysis. A genetic screen revealed that swrW is necessary for the hyper-hemolysis phenotype of crp mutants. The swrW gene is required for biosynthesis of the biosurfactant serratamolide, previously shown to be a broad-spectrum antibiotic and to contribute to swarming motility. Multicopy expression of swrW or mutation of the hexS transcription factor gene, a known inhibitor of swrW expression, led to an increase in hemolysis. Surfactant zones and expression from an swrW-transcriptional reporter were elevated in a crp mutant compared to the wild type. Purified serratamolide was hemolytic to sheep and murine red blood cells and cytotoxic to human airway and corneal limbal epithelial cells in vitro. The swrW gene was found in the majority of contact lens isolates tested. Genetic and biochemical analysis implicate the biosurfactant serratamolide as a hemolysin. This novel hemolysin may contribute to irritation and infections associated with contact lens use. © 2012 Shanks et al

Aspirin and its Metabolites Enhance the Expression of Vascular Endothelial Growth Factor in Retinal Pigment Epithelial Cell Cultures – Implications in the Pathophysiology of Age-Related Macular Degeneration

Purpose: An estimated 19.3% of adults, especially the elderly in the United States regularly use Aspirin for cardioprotection. Recently, multiple cohort studies have concluded that regular aspirin use for 10-15 years was associated with a statistically significant increase in the risk of incident age-related and neovascular acute macular degeneration. It has been hypothesized that aspirin or its metabolites induce the expression of vascular endothelial growth factor (VEGF). Materials & Methods: Retinal pigment epithelial cells, ARPE-19 (ATCC®CRL-2302™) were cultured. The cells were grown to achieve 95% confluence and then the media was changed. Cells cultured under blue light, red light, or darkness were subjected to a challenge with high dose aspirin (0.925 mg/dL), low dose aspirin (0.325 mg/dL), or hippuric acid (0.325 mg/dL). Light was generated using 2 red or blue LEDs powered by 3v CR2032 batteries. The 24-well plate was incubated with or without drugs in blue light, red light or darkness at 37C for 16 hours. The supernatants were harvested, and VEGF was quantified. One-way ANOVA using Dunnett’s multiple comparison test was performed to analyze statistical significance. Results: Cells exposed to blue light or darkness and hippuric acid showed a statistically significant increase in VEGF secretion (P=0.0012). However, cells exposed to red light with hippuric acid challenge showed no significant difference from the mean of cells exposed to darkness and sham control. Conclusions: Retinal pigment epithelial cells challenged with oxidative stress provided by blue light or darkness in the presence of hippuric acid increased VEGF secretion, suggesting a possible cause for neovascularization in age-related macular degeneration. RPE cells exposed to red light, known to abrogate oxidative stress, had decreased levels of VEGF induction by hippuric acid

Clinical outcomes of splenectomy in children: Report of the splenectomy in congenital hemolytic anemia registry

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/110704/1/ajh23888.pd