ARF regulating proteins as novel drug targets in Trypanosoma brucei

Human African Trypanosomiasis (HAT) and Animal African Trypanosomiasis (AAT) are neglected tropical diseases that pose a huge socioeconomic and health burden in poorer countries in sub-Saharan Africa. The causative agents of these diseases are the Trypanosoma brucei (T. brucei) spp., kinetoplastid parasites that is transmitted by tsetse flies (Diptera Glossina). Drugs against HAT and nagana are toxic, species and in nagana, host specific. With reports of emerging resistance to current drugs, it is essential to identify novel drug targets against T. brucei. This thesis focuses on identifying and characterising regulating proteins of ADP-ribosylation factors (ARFs) in T. brucei. ARFs are small GTPase binding proteins that have been shown to be essential in bloodstream form T. brucei. However the T. brucei ARFs share a high level of sequence identity with human ARFs, thus making them unsuitable as drug targets. Guanine nucleotide exchange factors (GEFs) and GTPase activating proteins (GAPs) regulate the activity of ARFs and have been shown to have highly diverse protein sequences. A total of 3 putative ARF GEFs and 4 putative ARF GAPs were identified in bloodstream form T. brucei by bioinformatics RNA interference (RNAi) cell lines were successfully generated for two GEFs and two GAPs in Lister 427 bloodstream form T. brucei. Tetracycline-induced knockdown was used to demonstrate that TbGEF3 is essential for viability of bloodstream form T. brucei. A decrease in cell growth was observed from 24 hours post RNAi, correlating with a decrease in cell cycle progression and an increase in cell death. The ‘BigEye’ phenotype could also be seen from 24 hours post RNAi, and cells were identified to have an endocytosis defect. These results combined with low level of sequence similarities at the essential ARF binding regions of TbGEF3 demonstrated that TbGEF3 may be a potential drug target. The TbGEF3 RNAi cells were used to assess the sensitivity of suramin, a drug taken up via endocytosis, in the presence of a partial endocytosis defect. Cells incubated in tetracycline for 24 hours prior to drug treatment had a reduced sensitivity to suramin compared to cells grown in the absence of tetracycline. The Pathogen Box set of compounds was screened for molecules with activity against T. brucei parasites. Identified hit compounds were used in preliminary studies to initiate development of an assay to distinguish compounds that are taken up via endocytosis. In summary, the putative ARF-regulating protein TbGEF3 has been shown to be essential for viability in T. brucei and has promise as a potential drug target. Further work is required in order to validate the use of the TbGEF3 RNAi line as a drug screening tool

Environmental Identity: A New Approach to Understanding Students' Participation in Environmental Learning Programs

The goal of this study is to develop an understanding of how participants express their environmental identities during an environmental learning program. Past research on the outcomes of environmental learning programs has focused primarily on changes in knowledge and attitudes. However, even if knowledge or attitudes can be accurately measured, they do not necessarily directly predict outcomes of environmental learning programs but rather serve as a proxy. Environmental identity is proposed as an alternative way to understanding participants' experiences in an environmental learning program. This study borrows a theoretical framework for environmental identity from the field of environmental campaigning and applies the field of environmental learning in a new way. This framework is based on known aspects of human identity from psychological research and allows environmental identity to be explored in a new and unique manner. A thematic analysis approach allows for the description of how six middle school students express their environmental identities around the themes of values and life goals, group membership, and fears and threats related to environmental issues. Findings indicate that the environmental identities of the participants in this study appear to be continuously developing and changing to account for new experiences and ideas related to the environment. Over the course of the program, some of the participants expressed that their environmental identities had strengthened due to their participation in the environmental learning program. The theoretical framework is used to describe what a strong environmental identity would look like as well as to characterize three of the participants' environmental identities. The affordances and limitations of the framework are shared and suggestions are made for how the framework could be strengthened for use in understanding participants' environmental identities during environmental learning programs

Recent advances in the surgical management of rhinosinusitis [version 1; referees: 4 approved]

Rhinosinusitis affects a significant portion of the US population, and its management imposes a substantial burden on the healthcare system. The treatment of chronic rhinosinusitis includes initial medical management prior to consideration of surgical intervention. However, if surgery does become necessary, several factors must be considered in order to optimize outcomes. This review evaluates surgical patient selection, perioperative medical management, and the extent of operative intervention, with the goal of improving surgical results, decreasing the need for revision surgery, and enhancing the patient’s quality of life. Specific variations in patient genotypes and phenotypes will be further explored with regard to their implications on surgical outcomes. Additionally, the evidence behind pre- and post-operative antibiotic and steroid use will be evaluated. Finally, we will review evolving surgical tools and techniques that are currently being utilized for the treatment of specific subsets of rhinosinusitis

Exploring RNNs for analyzing Zeek HTTP data

Cyber vulnerabilities pose a threat across systems in the Department of Defense. Finding ways to analyze network traffic and detect malicious behavior on a network will help keep these systems safe. This poster looks at the data collection techniques, model creation, and results of building a recurrent neural network to classify incoming traffic as normal or malicious. Additionally, it considers how the information will be best portrayed on a GUI to network administrators. The model\u27s initial accuracy is 83.45% when trained on 500,017 connections. With increased accuracy, this tool may be used by the Department of Defense to help defend its networks

Neuro-Ophthalmic Injuries With Systemic Neurologic Injury or Traumatic Brain Injury in Operation Iraqi Freedom and Operation Enduring Freedom

Background: To analyze the final visual acuity (VA) of neuro-ophthalmologic injuries (NOI) in combat ocular trauma and to study the association of NOI with systemic neurologic injury (SNI) and traumatic brain injury (TBI). Methods: The Walter Reed Ocular Trauma Database was reviewed. Inclusion criteria were any US service member and Department of Defense civilians who suffered NOI, SNI, or TBI in Operation Iraqi Freedom or Operation Enduring Freedom. The primary outcome measure was the rate of poor final VA in patients with an NOI with secondary outcome measures the rate of SNI and TBI. in patients with NOI. Results: One hundred seventy-eight (20.00%) of 890 eyes had an NOI. Optic nerve injury was observed in 79 (44.38%) eyes, other cranial nerve injury in 68 (38.20%), Horner syndrome in 4 (2.25%), diplopia in 45 (25.28%), and ptosis in 13 (7.30%). In patients with NOI, 76 (42.69%) eyes had a final VA less than 20/200. In injured eyes (n = 359) of patients (n = 251) with TBI, 154 eyes (34.26%) had a final VA less than 20/200. In multivariate analysis, optic nerve injury (P < 0.001), unlike TBI (P = 0.47), was associated with final VA less than 20/200. SNI (n = 229) had a statistically significant association (odds ratio 29.8, 95% confidence interval 19.2-47.8, P < 0.001) with NOI. Optic nerve injury and cranial nerve injury were associated with TBI and SNI (all, P < 0.001). Conclusion: Optic nerve injury and cranial nerve injury are associated with TBI and SNI. Optic nerve injury, but not TBI or cranial nerve injury, is associated with a poor final VA

Comparing the Management of Eye Injuries by Coalition Military Surgeons during the Iraq and Afghanistan Conflicts.

PURPOSE To compare incidences, ocular injury types, and treatment performed on United States and United Kingdom military service members and host nation civilians within the Iraq and Afghanistan conflicts to inform future military surgical training requirements and military medical planning. The United States routinely deployed ophthalmologists, whereas the United Kingdom did not. DESIGN Retrospective cohort study of the United States and United Kingdom military Joint Theatre Trauma Registries. PARTICIPANTS All patients with eye injuries treated at a deployed Military Treatment Facility between March 2003 and October 2011. METHODS An adjusted multiple logistic regression model was performed using enucleation or evisceration and primary open-globe repair as dependent variables and casualty nationality, location, and the presence of an ophthalmic surgeon as independent variables. MAIN OUTCOME MEASURES Incidence of eye removal (enucleation or evisceration) or primary repair for open globe injury. RESULTS Five thousand seven hundred nineteen of 67 586 (8%) survivors or those who died of wounds were recorded to have sustained eye injuries. The most common eye injuries were open-globe injury without intraocular foreign body (3201/5719 [56%]). Adnexal injuries (eyelid lacerations and damage to lacrimal apparatus) were recorded in 1265 of 5719 patients (22%). The odds of undergoing evisceration or enucleation for open-globe injury was highest in host nation civilians (odds ratio [OR], 9.23; P < 0.001), but there was no evidence of a difference between United States and United Kingdom military service member casualties (P = 0.38). The presence of an ophthalmic surgeon (OR, 16.3; P < 0.001) significantly affected the odds of eye removal. CONCLUSIONS Eye injuries were more likely to have been treated definitively in United States Medical Treatment Facilities (MTFs), reflecting the absence of ophthalmologists in most deployed United Kingdom MTFs. The Iraq and Afghan conflicts were notable for coalition air dominance; the shape of future conflicts may mandate delays in evacuation, which may affect visual outcomes negatively, particularly if primary repair of patients with open-globe injuries is delayed. This study provides evidence to support the maintenance of specialist ophthalmic surgical competencies in deployed coalition MTFs for future conflicts

Severity Assessment in CDKL5 Deficiency Disorder

BACKGROUND: Pathologic mutations in cyclin-dependent kinase-like 5 cause CDKL5 deficiency disorder, a genetic syndrome associated with severe epilepsy and cognitive, motor, visual, and autonomic disturbances. This disorder is a relatively common genetic cause of early-life epilepsy. A specific severity assessment is lacking, required to monitor the clinical course and needed to define the natural history and for clinical trial readiness. METHODS: A severity assessment was developed based on clinical and research experience from the International Foundation for CDKL5 Research Centers of Excellence consortium and the National Institutes of Health Rett and Rett-Related Disorders Natural History Study consortium. An initial draft severity assessment was presented and reviewed at the annual CDKL5 Forum meeting (Boston, 2017). Subsequently it was iterated through four cycles of a modified Delphi process by a group of clinicians, researchers, industry, patient advisory groups, and parents familiar with this disorder until consensus was achieved. The revised version of the severity assessment was presented for review, comment, and piloting to families at the International Foundation for CDKL5 Research-sponsored family meeting (Colorado, 2018). Final revisions were based on this additional input. RESULTS: The final severity assessment comprised 51 items that comprehensively describe domains of epilepsy; motor; cognition, behavior, vision, and speech; and autonomic functions. Parental ratings of therapy effectiveness and child and family functioning are also included. CONCLUSIONS: A severity assessment was rapidly developed with input from multiple stakeholders. Refinement through ongoing validation is required for future clinical trials. The consensus methods employed for the development of severity assessment may be applicable to similar rare disorders