Demyelination in the CNS is known to involve several immune effector mechanisms, including complement proteins. For this dissertation project the central hypothesis that C3 and downstream effector complement proteins exacerbate demyelination through activation of glial cells was tested. To investigate the role of C3 and downstream complement proteins in demyelination and remyelination pathology in vivo we utilized the cuprizone model. We used C3 knockout mice (C3-/-), which are lacking the central C3 protein and subsequently all downstream complement effector proteins, and transgenic mice expressing C3a or C5a under the control of the glial GFAP promoter. Interestingly, we found no changes in demyelination or remyelination pathology between C3-/- and control mice. However, C3a and C5a transgenic mice had exacerbated demyelination and slightly delayed remyelination in the corpus callosum compared to WT mice. Transgenic mice had increased cellularity in the corpus callosum due to increased activation and/or migration of microglia. There was also evidence of T cells in the corpus callosum during demyelination in C5a transgenic mice, suggesting C5a may modulate BBB permeability. During early remyelination oligodendrocytes migrated to the corpus callosum in higher numbers in C3a and C5a transgenic mice, thus enabling these mice to remyelinate as effectively as WT mice by the end of the ten week study. To determine the effects of anaphylatoxins on individual glial subsets, we created murine recombinant C3a and C5a proteins. We found that the MAPK pathway proteins JNK1 and ERK1/2 were activated in glia upon stimulation with recombinant anaphylatoxin proteins. When microglia and mixed glial cultures were stimulated with C3a and/or C5a, we observed an increase in the production of proinflammatory cytokines and chemokines. In contrast, anaphylatoxin-treated primary astrocytes had suppressed cytokine and chemokine production compared to untreated astrocytes. In vitro, primary microglia and astrocytes did not significantly migrate in response to stimulation with C3a or C5a proteins, suggesting migration may not be a primary anaphylatoxin-mediated function in the CNS. Overall, our findings show that anaphylatoxin production in the brain plays a negative proinflammatory role during demyelination and that anaphylatoxin proteins can activate individual subsets of glia, initiating the production of inflammatory mediators.

Thesis (M.S.)--University of Delaware, 2006. / Principal faculty advisor: Cheng-Shun (Richard) Fang, Dept. of Health, Nutrition, and Exercise Sciences. Includes bibliographical references.

Multiple Sclerosis (MS) is the most common demyelinating disease affecting the central nervous system. Over 80% of MS patients are in the relapsing remitting stage. Symptoms range from fever, fatigue, emotional distress, tingling, numbness, optic neuritis, spasticity, muscle weakness, impaired coordination, to other abnormal neurological problems. Expression of symptoms is known as a relapse or exacerbation. The cause of relapses is unknown, but multiple factors seem to play a significant role. Possible factors that may influence MS onset and relapse consist of a genetic association, viruses, disruption of the blood-brain barrier, and stress. Stress has shown to have negative implications and may stimulate relapses. Thus, this study examined a possible stress intervention that most people already had available to them, companion animals. Companion animals have been shown to lower blood pressure, decrease heart rate, provide social support, and reduce stress. The main hypothesis was to evaluate whether or not pet ownership and/or attachment influenced the perceived stress level and number of negative life events experienced by MS patients in the relapsing remitting stage. Participants were given a questionnaire that consisted of 7 surveys. The questionnaire accessed quality of life, disease severity, number of negative life events, perceived stress level, level of depression, social support, and pet ownership and attachment level. Our sample population consisted of MS patients seen at the University of Texas Southwestern Neurology clinic from February 23rd to May 21st, 2004. One hundred and forty seven relapsing remitting MS patients were included in the study. Multiple linear regression was used to compare the relationship of stress and number of negative life events to pet ownership and attachment. Results revealed that pet ownership and attachment levels did not affect the stress level and number of negative life events of MS patients. No confounders were identified. Interaction terms with disease severity as the dependent variable, pet ownership and perceived stress level or negative life events as the independent variables were not significant. The type of pet owned did not influence the attachment level of the MS patient. In conclusion, the results of this study did not support the hypothesis.

The effects of multiple sclerosis (MS) on the central nervous system often manifest as abnormalities in gait kinematics. Clinically practical, valid, and reliable measures of gait kinematics are necessary to address research and clinical questions about MS. Wireless flexible electrogoniometry (EG) is a clinically practical measure of joint angles. The GAITRite walkway system is a clinically practical, valid and reliable measure of temporal and spatial gait characteristics. The overall objective of this two-study research project was to explore whether these clinically practical measures of gait kinematics can be used to accurately detect gait abnormalities in people with multiple sclerosis. Study 1 examined the reliability and validity of EG and Study 2 examined the gait kinematics of people with MS (PWMS) using EG and GAITRite. For Study 1, angle at initial contact and total joint excursion were measured by EG at both the knee and ankle while ten healthy adults walked at a self-selected comfortable speed. Measurements were repeated for two testers and two visits to assess reliability. The same variables were measured concurrently with three-dimensional motion analysis (3D) to assess validity. For all variables, reliability was good as indicated by low measurement error and validity was good as indicated by association and agreement of EG with 3D. For Study 2, the same joint angles, along with speed, cadence, step length, stride length, stance duration and double support duration were assessed for six PWMS and six controls without MS. PWMS showed significantly reduced speed, cadence, and ankle excursion and increased stance and double support duration as previously shown with 3D. Spasticity and/or instability may lead to these kinematic gait abnormalities in PWMS; however, reduced velocity may confound this interpretation by affecting the other observed gait abnormalities. Further research about the determinants of gait dysfunction in PWMS is required. EG and GAITRite are clinically practical, valid and reliable measures of gait kinematics and should be included in further clinic-based research to determine which kinematic gait abnormalities are causes and which are effects of the observed decrease in gait speed in PWMS.

Cognitive impairment affects approximately half of multiple sclerosis (MS) patients. The Multiple Sclerosis Neuropsychological Questionnaire (MSNQ) has previously demonstrated validity for detecting cognitive impairment in MS, and is quick and easy to complete. The objective was to validate an internet version of the MSNQ. The following were completed at home over the internet for 82 MS patients: (a) patient self-report version of the MSNQ (P-MSNQ), (b) informant version of the MSNQ (I-MSNQ), and (c) Centre for Epidemiological Studies Depression Scale (CES-D). Thereafter, patients completed in-office neuropsychological testing using the Brief Repeatable Battery of Neuropsychological Tests (BRB-N). Both the P-MSNQ and I-MSNQ were highly correlated with depression. The best-cut off score on the I-MSNQ was a 26, which gave a sensitivity of 72% and 60% for detecting cognitive impairment on the BRB-N. Given the modest sensitivity and specificity values, the MSNQ is not recommended for neuropsychological screening purposes over the internet.

Individuals with Multiple Sclerosis commonly use assistive mobility devices, such as rollators, to compensate for their mobility impairments. However, the effect of these devices on their foot placement during gait has not been explored in the community. The objective of experiment one was to develop and validate a tool that quantifies medio-lateral foot placement characteristics during rollator use. In this study, a technique was developed for an instrumented rollator (i.e. iWalker) and validated against a Vicon motion capture system in able-bodied young adults. The two systems were in strong agreement. The objective of experiment two was to apply this iWalker-based technique to individuals with Multiple Sclerosis to identify environment-related foot placement changes. This study revealed that step width variability, but not step width, can be influenced by certain outdoor environments. Therefore, environmental context is important to consider when investigating user-device interactions and factors responsible for safe mobility in this population.

Cognitive impairment affects approximately half of multiple sclerosis (MS) patients. The Multiple Sclerosis Neuropsychological Questionnaire (MSNQ) has previously demonstrated validity for detecting cognitive impairment in MS, and is quick and easy to complete. The objective was to validate an internet version of the MSNQ. The following were completed at home over the internet for 82 MS patients: (a) patient self-report version of the MSNQ (P-MSNQ), (b) informant version of the MSNQ (I-MSNQ), and (c) Centre for Epidemiological Studies Depression Scale (CES-D). Thereafter, patients completed in-office neuropsychological testing using the Brief Repeatable Battery of Neuropsychological Tests (BRB-N). Both the P-MSNQ and I-MSNQ were highly correlated with depression. The best-cut off score on the I-MSNQ was a 26, which gave a sensitivity of 72% and 60% for detecting cognitive impairment on the BRB-N. Given the modest sensitivity and specificity values, the MSNQ is not recommended for neuropsychological screening purposes over the internet.

Individuals with Multiple Sclerosis commonly use assistive mobility devices, such as rollators, to compensate for their mobility impairments. However, the effect of these devices on their foot placement during gait has not been explored in the community. The objective of experiment one was to develop and validate a tool that quantifies medio-lateral foot placement characteristics during rollator use. In this study, a technique was developed for an instrumented rollator (i.e. iWalker) and validated against a Vicon motion capture system in able-bodied young adults. The two systems were in strong agreement. The objective of experiment two was to apply this iWalker-based technique to individuals with Multiple Sclerosis to identify environment-related foot placement changes. This study revealed that step width variability, but not step width, can be influenced by certain outdoor environments. Therefore, environmental context is important to consider when investigating user-device interactions and factors responsible for safe mobility in this population.

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 effects of multiple sclerosis (MS) on the central nervous system often manifest as abnormalities in gait kinematics. Clinically practical, valid, and reliable measures of gait kinematics are necessary to address research and clinical questions about MS. Wireless flexible electrogoniometry (EG) is a clinically practical measure of joint angles. The GAITRite walkway system is a clinically practical, valid and reliable measure of temporal and spatial gait characteristics. The overall objective of this two-study research project was to explore whether these clinically practical measures of gait kinematics can be used to accurately detect gait abnormalities in people with multiple sclerosis. Study 1 examined the reliability and validity of EG and Study 2 examined the gait kinematics of people with MS (PWMS) using EG and GAITRite. For Study 1, angle at initial contact and total joint excursion were measured by EG at both the knee and ankle while ten healthy adults walked at a self-selected comfortable speed. Measurements were repeated for two testers and two visits to assess reliability. The same variables were measured concurrently with three-dimensional motion analysis (3D) to assess validity. For all variables, reliability was good as indicated by low measurement error and validity was good as indicated by association and agreement of EG with 3D. For Study 2, the same joint angles, along with speed, cadence, step length, stride length, stance duration and double support duration were assessed for six PWMS and six controls without MS. PWMS showed significantly reduced speed, cadence, and ankle excursion and increased stance and double support duration as previously shown with 3D. Spasticity and/or instability may lead to these kinematic gait abnormalities in PWMS; however, reduced velocity may confound this interpretation by affecting the other observed gait abnormalities. Further research about the determinants of gait dysfunction in PWMS is required. EG and GAITRite are clinically practical, valid and reliable measures of gait kinematics and should be included in further clinic-based research to determine which kinematic gait abnormalities are causes and which are effects of the observed decrease in gait speed in PWMS.