Biocuration is manual scientific collection, annotation and validation of literary information of biological and model organisms into a single database. Successful biocuration processes involve those with an extensive collection of literature, a user-friendly database interfaces for entering and analyzing data from published papers, and highly regulated training and quality assurance protocols. Due to the rapid expansion of biomedical literature, an efficient and accurate biocuration process has become more valuable due to the magnitude of data available in published literature. As the biocuration process incorporates undergraduates, it is critical that the medium for data collection is simple, ergonomic, and infallible. A reconstructed FileMaker Pro database was introduced to previously trained undergraduate students for process evaluation. Streamlining the biocuration process and grouping data structure to be more intuitive were two goals the new database interface hoped to achieve. The creation of a rigorous training program and strict quality management protocol is needed to prepare the lab for the introduction of efficient biocuration processes. Through the database designing process, training protocols were drafted to effectively call the biocurator’s attention to important changes in the interface design. Upon prototyping the database, entry errors were reviewed, training protocols were adjusted, and the quality protocols were drafted. When the combination of undergraduate biocurators and the reconstructed database under these new protocols was compared to statistics in the biocuration field, results proved to show increase in both productivity rates as well as accuracy rates. By having such efficiency at the undergraduate level, subject matter experts will no longer be required to perform this type of research and can focus on analysis. This will increase research productivity and reduce costs in the overall biocuration process. With over 12,000 published papers regarding Amyotrophic Lateral Sclerosis on Pubmed in 2014 alone, this revolutionary combination could lead to quickly finding a suitable cure for these patients.

published_or_final_version / Zoology / Master / Master of Philosophy

No description available.

Interactions between the immune system and the nervous system are currently underappreciated, assumed to play minor mechanistic roles in disease pathogenesis. In contrast, our laboratory has demonstrated the importance of this relationship with significant impact, initially in Type 1 Diabetes (T1D). The experiments presented here build on our previous work to provide insights into the etiology of Multiple Sclerosis (MS) and Type 2 Diabetes (T2D). Transient Receptor Potential Vanilloid-1 (TRPV1) is an ion channel expressed on peripheral sensory afferent neurons that fundamentally control T1D pathogenesis. Here we show that mice with genetic ablation of TRPV1 are protected from EAE progression, attributable to reduced central nervous system (CNS) leukocyte passage. The pathogenic role of TRPV1 in permeabilizing the blood-CNS barriers may also translate to MS, as patients with progressive disease show a significant mutation bias within the TRPV1 gene. We were simultaneously intrigued by the growing worldwide obesity epidemic, and we observed that obese mice develop more severe EAE compared to lean mice. This was mechanistically linked to an expansion of TH17 cells, driven by sustained rises of IL-6 in obese mice. This research implies new therapeutic opportunities for the many obese patients with diverse autoimmune diseases. Finally, the immune system, obesity, and T2D are functionally linked, and we contributed to research that uncovered a large presence of immune cells in adipose tissue that drive insulin resistance. Manipulation of T cells and B cells affects local inflammation as well as whole-body insulin resistance and glucose homeostasis. Intriguingly, auto-antibodies in insulin resistant individuals are specific for a number of unique proteins, including glial fibrillary acidic protein (GFAP), initially shown by our laboratory to play a key role in T1D progression. We further characterized the autoimmune and neuronal progression elements that steer disease pathogenesis, and observed that administration of a vaccine containing GFAP is able to dramatically reduce weight gain and insulin resistance in mice. The data presented in this thesis provide a number of novel, mechanistic observations linking the immune and nervous systems in disease, and implies several potential avenues for treatment.

Recent reviews (Peyser & Poser, 1986; Rao, 1986) suggest that Multiple Sclerosis results in cognitive impairments in the areas of learning and memory, abstract reasoning, information processing efficiency, and, often, visual-spatial ability. Whether this pattern applies to the individual with MS is unclear. Due to the disseminated distribution of MS neuropathology, patients may undergo idiosyncratic cognitive changes dependent upon the site of white matter lesions. The present study explored this question using cluster analysis on the neuropsychological data from a group of mildly disabled MS patients (n = 177) and a well-matched control group (n=89). In a group of MS patients who were identified with unequivocal cognitive impairment, the resultant clusters indicated that MS does not result in a characteristic pattern of impairment. Two clusters were obtained that resembled the pattern described in the literature, while the majority of patients clustered into groups with specific deficits in one or two areas, with normal performance in others. In order to identify associations between cluster groups and lesion sites, frequency tables were constructed for the presence of a lesion on Magnetic Resonance Imaging in 24 brain sites. An association was obtained between two lesion sites and two cognitive tests. Visual-spatial impairment, as assessed by the Benton Visual Retention test, was associated with lesions in the genu of the corpus callosum and with more lesions throughout the corpus callosum. Impaired performance on Paired Associates, a test of learning and memory for novel verbal associations, was associated with a lesion in the deep white matter of the left parietal lobe. The results support the hypothesis that MS results in multiple patterns of cognitive impairment depending on the individual placement of white matter lesions. Identifying and characterizing the heterogeneity of the impairment may greatly increase our understanding of the role of myelin in cognition and the functions of white matter tracts in the brain.

T2 relaxation imaging can be used to measure signal from water trapped between myelin bilayers; the ratio of myelin water signal to total water is termed the myelin water fraction (MWF). First, results from multi-component T2 relaxation and diffusion tensor imaging (DTI) were compared for 19 multiple sclerosis (MS) subjects at 1.5 T to better understand how each measure is affected by pathology. In particular, it was determined that the detection of a long-T2 signal within an MS lesion may be indicative of a different underlying pathology than is present in lesions without long-T2 signal. Next, the single-slice T2 relaxation measurement was implemented, refined, and validated at 3.0 T. Scan parameters were varied for phantoms and in-vivo brain, and changes in multi-exponential fit residuals and T2 distribution-derived parameters such as MWF were monitored to determine which scan parameters minimized artifacts. Measurements were compared between 1.5 T and 3.0 T for 10 healthy volunteers. MWF maps were qualitatively similar between field strengths. MWFs were significantly higher at 3.0 T than at 1.5 T, but with a strong correlation between measurements at the different field strengths. Due to long acquisition times, multi-component T2 relaxation has thus far been clinically infeasible. The next study aimed to validate a new 3D multi-component T2 relaxation imaging technique against the 2D single-slice technique most commonly used. Ten healthy volunteers were scanned with both the 2D single-slice and 3D techniques. MWF maps were qualitatively similar between scans. MWF values were highly correlated between the acquisition methods. Although MWF values were generally lower using the 3D technique, they were only significantly so for peripheral brain structures, likely due to increased sensitivity of slab-selective refocusing pulses used for the 3D approach. The 3D T2 relaxation sequence was then applied to the study of MS to take advantage of the increased brain coverage. Thirteen MS subjects and 11 controls underwent T2 relaxation and DTI examinations to produce histograms of MWF and several DTI-derived metrics. MS MWF histograms differed considerably from those of controls, and differences in MS MWF histograms did not mirror differences in DTI histograms relative to matched controls.

The geographic disparity of multiple sclerosis has been noted in the literature for well over a century. The frequency of the disease varies significantly both within countries and in different parts of the world. The goal of this project is to give new insight regarding the etiology of multiple sclerosis. Several theories regarding the etiology of the disease have been reviewed, including a geographic theory, a nutritional theory, and a genetic theory. Although the geographic and nutritional theories have been thoroughly investigated by researchers, neither of them provides a conclusive explanation for the etiology of the disease, and there are discrepancies with respect to both theories. The purpose of this study is to reveal the discrepancies in the epigenetic theories regarding the etiology of multiple sclerosis and to demonstrate the correlation of multiple sclerosis prevalence and the migration and settlement history of Northern Europeans, thus conferring the passage of a genetic susceptibility to the disease.

Multiple sclerosis (MS) is an inflammatory autoimmune disease of the central nervous system. We hypothesize that angiogenesis results in new vessels which serve as a conduit for immune cell recruitment in MS, and contribute to inflammation through the pro-inflammatory properties of angiogenic regulators. This study is one of the first to explore regulation of angiogenesis in a murine model of MS (experimental autoimmune encephalomyelitis, EAE), and its potential as a therapeutic target. Angiogenesis was apparent 21 days following disease induction and correlated with clinical and pathologic scores. We documented alterations in the VEGF and Ang/Tie signaling pathways. Expression of VEGF increased at day 14 then reduced by day 21. At this time point, Ang-2 levels were elevated due to expression by infiltrating macrophages. Ang-1 was significantly reduced at day 14 and increased at day 21 due to expression by CD3+ T-cells. The same expression pattern was validated in inflammatory cells within human MS tissue. Vascular permeability increased at day 14 and returned to control levels at day 21. The volume of permeable tissue weakly correlated with angiogenesis. VEGF blockage with bevacizumab suppressed angiogenesis and reduced clinical scores and vascular permeability. Retention of CD4+ T-cells in peripheral lymph nodes and reduced T-cell proliferation was noted following treatment. Bevacizumab reduced mononuclear cell infiltration into spinal cord. Isolated CD4+ T-cells showed reduced expression of IL-17 and IFN-??. B20-4.1.1 (a monoclonal antibody against murine VEGF) reduced clinical scores and suppressed angiogenesis as did treatment with angiostatin (an inhibitor of endothelial cell proliferation). B20-4.1.1 reduced vascular permeability, induced retention of CD4+ T-cells in peripheral lymph nodes, and inhibited T-cell proliferation, while angiostatin had no effect. Isolated lymphoid cells from mice treated with both agents showed reduced secretion of IL-17, but B20-4.1.1 had no effect on Tcell proliferation or IL-17 secretion when combined with angiostatin. ?? We conclude that these angiogenesis inhibitors are effective in EAE and act by suppressing angiogenesis with a secondary effect on peripheral T-cell activation. To the extent that EAE replicates changes occurring in MS, we have demonstrated that modulation of angiogenesis may represent a promising strategy in the management of disease progression.

Butyrylcholinesterase (BuChE) is a serine hydrolase enzyme that, along with acetylcholinesterase (AChE), catalyzes the hydrolysis of acetylcholine. These enzymes are associated with the pathology of neurologic disorders such as Alzheimer's disease (AD) and multiple sclerosis (MS). In particular, AChE and BuChE accumulate in B- amyloid (AB) plaques and tau neurofibrillary tangles in the AD brain. Thus, imaging cholinesterase activity associated with plaques and tangles in the brain has the potential to provide definitive diagnosis of AD during life. This would be advantageous since, at present, confirmation of AD relies on detecting pathology through post-mortem examination of the brain. In a similar respect, BuChE is associated with the characteristic lesions in MS brain and thus, is a promising target for diagnosis and monitoring of pathology in this disease. It is hypothesized that cholinesterase-binding radiopharmaceuticals can be used in SPECT or PET imaging to visualize these enzymes associated with AD and MS pathology in the living brain. Several classes of cholinesterase ligands were synthesized and exhibited potent binding and specificity towards AChE and BuChE using enzyme kinetic analysis. These compounds were rapidly radiolabelled with 123I and purified. Radiolabelled molecules accumulated in vitro in areas known to contain cholinesterase activity in transgenic AD mice and post-mortem human AD brain tissues, using autoradiography. Furthermore, cholinesterase activity associated with AB plaques was visualized in human and transgenic mouse AD brain tissues. An enzyme kinetic approach was employed to determine critical residues in the BuChE active site gorge for ligand binding. In particular, residues pertaining to the peripheral site of the enzyme were identified and found to be involved in the binding of various ligands. These results are crucial for optimizing the enzyme binding properties of cholinesterase imaging agents. Finally, PET imaging of a transgenic mouse model of AD was performed as a vanguard for pre-clinical evaluation of cholinesterase imaging agents. PET imaging identified similar characteristics between this AD mouse model and the human condition. This is a promising approach for evaluation of cholinesterase imaging agents. Radioligands specific for cholinesterases have the potential to provide a noninvasive means for early diagnosis of neurological diseases using brain scanning.

Cognitive deficits are highly prevalent in multiple sclerosis (MS) and have a negative impact on daily life. Impairments in information processing speed are among the most commonly reported deficits in MS and are generally assessed by evaluating mean-level performance on time-limited tests. However, this approach to assessing performance ignores potential within-subject differences that may be useful for characterizing cognitive difficulties in MS. An alternative method of measuring performance on timed cognitive tasks is to examine the degree of within-subject variability, termed intraindividual variability (IIV). IIV provides information about the characteristics of an individual’s performance and may provide novel information about cognitive functioning in MS and other neurodegenerative disorders. The research presented in this dissertation examined IIV in performance as an indicator of cognitive functioning in persons with MS and explored the relations of performance variability to measures of neuronal connectivity derived from resting state functional magnetic resonance imaging (rsfMRI). Individuals with MS were found to be both slower and more variable on tests of information processing speed and attention. This variability was observed even when controlling for sensorimotor confounds and other systematic variables that may influence variability, such as practice and learning effects. IIV in performance was found to better distinguish MS patients from matched groups of healthy control subjects when compared to common clinical measures of cognitive performance or average response speed. These differences in IIV were also found consistently across six monthly assessments in a group with MS who remained clinically stable over this period. This stability in IIV suggests its feasibility as a measure of changes in longitudinal cognitive or clinical status. Using rsfMRI, greater stability in performance (i.e., lower IIV) was associated with greater functional connectivity between frontal lobe regions (i.e., ventral medial prefrontal cortex and frontal pole) in persons with MS. This increased connectivity appears to represent potential compensatory processes within mildly affected MS individuals. Together the findings demonstrate that IIV is an important characteristic of cognitive performance that may provide new insights into the cognitive deficits present in MS.