Investigating in vivo Brain Metabolite Levels in Concussed Female Athletes and a Murine Model of Repetitive Closed Head Injury

After a concussion there is a complex cascade of events, termed the neurometabolic cascade, that includes changes in ion flux, neurotransmission, and cellular energetics. How this pathophysiological process translates into cognitive deficits remains poorly understood. Magnetic resonance spectroscopy (MRS) provides a non-invasive technique that allows for the quantification of brain metabolites that are involved in these processes, including glutamate and glutamine, which are involved in neurotransmission. Moreover, female athletes are underrepresented in studies on concussion, limiting our knowledge and understanding of sex differences. The overall goal of this thesis was to examine metabolite changes using MRS in female athletes before and after concussion, with the added goal of quantifying glutamate and glutamine separately. The second objective was to replicate metabolite changes in an animal model of concussion, to position future studies to probe the reasons for these changes, and to explore whether these changes represent potential therapeutic targets. MRS was acquired from the prefrontal WM of female athletes (contact and non-contact sports) and sedentary women at 3T to explore metabolite differences between groups and changes after concussion. In addition, an animal model of repeated closed head impacts was studied at 9.4T, in an effort to replicate the findings observed in humans. In the contact athlete cohort, reduced glutamine and glutamine/total creatine (Gln/Cr) were found following concussion, and after a season of play in non-concussed athletes. In the non-contact athlete cohort, metabolite levels did not change over the course of a season, and they did not differ from age matched sedentary women, except for a small difference in myo-inositol. Most interestingly, glutamine levels were significantly elevated in contact athletes compared to sedentary and non-contact groups, suggesting that sub-concussive impacts may have a long-term effect on brain metabolite levels. Furthermore, the large difference in glutamine levels between contact and non-contact athletes has implications in study design in regards to control groups versus test-retest paradigm. In the final study, we used a murine model (C57BL6) of repeated closed head injury to investigate metabolite level changes post-injury. Elevated Gln/Cr was observed 10-weeks post-injury, suggesting that the model may be appropriate to study sub-concussive injury. Together, these studies suggest that there exists a cumulative effect on the brain from sub-concussive impacts in contact sports, that manifests as elevated glutamine levels. Moreover, concussion in the same cohort of athletes results in reduced glutamine levels. Further work aimed at replicating these findings in animal models will be crucial to understanding the effects of cumulative impacts and concussion

Brain Metabolite Levels in Sedentary Women and Non-contact Athletes Differ From Contact Athletes

White matter tracts are known to be susceptible to injury following concussion. The objective of this study was to determine whether contact play in sport could alter white matter metabolite levels in female varsity athletes independent of changes induced by long-term exercise. Metabolite levels were measured by single voxel proton magnetic resonance spectroscopy (MRS) in the prefrontal white matter at the beginning (In-Season) and end (Off-Season) of season in contact (N = 54, rugby players) and non-contact (N = 23, swimmers and rowers) varsity athletes. Sedentary women (N = 23) were scanned once, at a time equivalent to the Off-Season time point. Metabolite levels in non-contact athletes did not change over a season of play, or differ from age matched sedentary women except that non-contact athletes had a slightly lower myo-inositol level. The contact athletes had lower levels of myo-inositol and glutamate, and higher levels of glutamine compared to both sedentary women and non-contact athletes. Lower levels of myo-inositol in non-contact athletes compared to sedentary women indicates long-term exercise may alter glial cell profiles in these athletes. The metabolite differences observed between contact and non-contact athletes suggest that non-contact athletes should not be used as controls in studies of concussion in high-impact sports because repetitive impacts from physical contact can alter white matter metabolite level profiles. It is imperative to use athletes engaged in the same contact sport as controls to ensure a matched metabolite profile at baseline

Genomic investigations of unexplained acute hepatitis in children

Since its first identification in Scotland, over 1,000 cases of unexplained paediatric hepatitis in children have been reported worldwide, including 278 cases in the UK1. Here we report an investigation of 38 cases, 66 age-matched immunocompetent controls and 21 immunocompromised comparator participants, using a combination of genomic, transcriptomic, proteomic and immunohistochemical methods. We detected high levels of adeno-associated virus 2 (AAV2) DNA in the liver, blood, plasma or stool from 27 of 28 cases. We found low levels of adenovirus (HAdV) and human herpesvirus 6B (HHV-6B) in 23 of 31 and 16 of 23, respectively, of the cases tested. By contrast, AAV2 was infrequently detected and at low titre in the blood or the liver from control children with HAdV, even when profoundly immunosuppressed. AAV2, HAdV and HHV-6 phylogeny excluded the emergence of novel strains in cases. Histological analyses of explanted livers showed enrichment for T cells and B lineage cells. Proteomic comparison of liver tissue from cases and healthy controls identified increased expression of HLA class 2, immunoglobulin variable regions and complement proteins. HAdV and AAV2 proteins were not detected in the livers. Instead, we identified AAV2 DNA complexes reflecting both HAdV-mediated and HHV-6B-mediated replication. We hypothesize that high levels of abnormal AAV2 replication products aided by HAdV and, in severe cases, HHV-6B may have triggered immune-mediated hepatic disease in genetically and immunologically predisposed children

Differential effects of transcranial direct current stimulation on antiphase and inphase motor tasks: A pilot study

© 2019 Elsevier B.V. Ageing is associated with a decline in motor function that critically interferes with activities of daily living involving manual dexterity. Transcranial direct current stimulation (tDCS) is a form of non-invasive brain stimulation that has been shown to enhance manual dexterity in healthy aging adults. The supplementary motor area (SMA) is involved in motor preparation and bimanual control; therefore, bihemispheric tDCS incorporating the SMA may preferentially enhance bimanual motor movements in healthy older adults. The aim of the current study was to determine if tDCS incorporating SMA could improve manual dexterity in older adults. Twenty-four adults, aged 67–84 participated in this double-blind, randomized, cross over design, pilot study. One group of participants (n = 17) were randomized to receive stimulation or sham on their first visit and received the contrary on their second visit, seven days later. A second group of participants (n = 10) received three consecutive days of tDCS while performing a motor task. Participants performed unimanual and bimanual hand movements while receiving 2 mA of tDCS. The total time for participants to complete three trials of each task was recorded. No significant differences in performance times were observed between single or tri session tDCS and sham conditions. However, tDCS had opposing effects on the motor consolidation of anti-phase and in-phase bimanual tasks. During the tri session paradigm, older adults improved performance learning of antiphase bimanual movements more quickly than inphase bimanual movements, suggesting a different mechanism of action of these two movements

Reduced Brain Glutamine in Female Varsity Rugby Athletes After Concussion and in Non-Concussed Athletes After a Season of Play

The purpose of this study was to use non-invasive proton magnetic resonance spectroscopy (MRS) and diffusion tensor imaging (DTI) to monitor changes in prefrontal white matter metabolite levels and tissue microstructure in female rugby players with and without concussion (ages 18-23, n = 64). Evaluations including clinical tests and 3 T MRI were performed at the beginning of a season (in-season) and followed up at the end of the season (off-season). Concussed athletes were additionally evaluated 24-72 hr (n = 14), three months (n = 11), and six months (n = 8) post-concussion. Reduced glutamine at 24-72 hr and three months post-concussion, and reduced glutamine/creatine at three months post-concussion were observed. In non-concussed athletes (n = 46) both glutamine and glutamine/creatine were lower in the off-season compared to in-season. Within the MRS voxel, an increase in fractional anisotropy (FA) and decrease in radial diffusivity (RD) were also observed in the non-concussed athletes, and correlated with changes in glutamine and glutamine/creatine. Decreases in glutamine and glutamine/creatine suggest reduced oxidative metabolism. Changes in FA and RD may indicate neuroinflammation or re-myelination. The observed changes did not correlate with clinical test scores suggesting these imaging metrics may be more sensitive to brain injury and could aid in assessing recovery of brain injury from concussion

Repetitive mild traumatic brain injury in mice triggers a slowly developing cascade of long-term and persistent behavioral deficits and pathological changes

We have previously reported long-term changes in the brains of non-concussed varsity rugby players using magnetic resonance spectroscopy (MRS), diffusion tensor imaging (DTI) and functional magnetic imaging (fMRI). Others have reported cognitive deficits in contact sport athletes that have not met the diagnostic criteria for concussion. These results suggest that repetitive mild traumatic brain injuries (rmTBIs) that are not severe enough to meet the diagnostic threshold for concussion, produce long-term consequences. We sought to characterize the neuroimaging, cognitive, pathological and metabolomic changes in a mouse model of rmTBI. Using a closed-skull model of mTBI that when scaled to human leads to rotational and linear accelerations far below what has been reported for sports concussion athletes, we found that 5 daily mTBIs triggered two temporally distinct types of pathological changes. First, during the first days and weeks after injury, the rmTBI produced diffuse axonal injury, a transient inflammatory response and changes in diffusion tensor imaging (DTI) that resolved with time. Second, the rmTBI led to pathological changes that were evident months after the injury including: changes in magnetic resonance spectroscopy (MRS), altered levels of synaptic proteins, behavioural deficits in attention and spatial memory, accumulations of pathologically phosphorylated tau, altered blood metabolomic profiles and white matter ultrastructural abnormalities. These results indicate that exceedingly mild rmTBI, in mice, triggers processes with pathological consequences observable months after the initial injury