Patients with temporal lobe epilepsy (TLE) often complain of a fading of new memories over days to weeks. This is particularly the case for patients with transient epileptic amnesia (TEA), a subtype of TLE. Objective memory testing sometimes corroborates this complaint, demonstrating normal or near-normal recall after standard delays (10-30 minutes), followed by a rapid decline in recall over longer delays (i.e. 1 week). This ‘nonstandard’ form of memory impairment has been termed accelerated long-term forgetting (ALF). It may reflect impairment of memory encoding, consolidation or retrieval. The aim of this thesis was to characterise the cognitive basis of ALF in TEA/TLE. The objectives were to: (a) determine the time scale of ALF of words (Chapter 3), (b) establish whether ALF affects picture recognition (Chapter 4), (c) establish whether ALF is affected by repeated retrieval (Chapter 2), number of learning trials (Chapter 5) and post-learning sensory stimulation (interference) (Chapter 5), (d) investigate ALF under incidental encoding conditions (Chapter 6), and (e) examine ALF associated with baclofen, a GABAB – receptor agonist (Chapter 7). A range of experimental paradigms and materials were applied to test memory function in several samples of TEA/TLE patients complaining of ALF and in healthy controls. The experiments revealed the following: ALF for word lists became apparent after 3–8 hours of daytime wakefulness, suggesting that disturbance of sleep related consolidation processes is not necessary for ALF to emerge in TEA. ALF for verbal information occurred both under incidental and intentional encoding conditions, and this rapid forgetting was not prevented by cued or recognition tests or by the matching of encoding conditions for patients and controls. This suggests that ALF is not associated primarily with an encoding or retrieval deficit. Although multiple learning trials and reduced sensory stimulation after learning reduced early forgetting (over 15-30 minutes) in TEA/TLE, neither factor reduced long-term forgetting. Moreover, in contrast to verbal recall, picture recognition was impoverished after minutes, but declined normally thereafter, demonstrating a subtle ‘early’ memory deficit in TEA, which might or might not be related to ALF. Overall, the present research suggests that ALF reflects a consolidation deficit, which results in accelerating forgetting the first few hours to days after memory acquisition, without a requirement for intervening sleep.

<p>Neurodevelopmental disorders develop over time and are characterized by a wide variety of mental, behavioral, and physical phenotypes. The categorization of neurodevelopmental disorders encompasses a broad range of conditions including intellectual disability, autism spectrum disorder, attention deficit hyperactivity disorder, cerebral palsy, schizophrenia, bipolar disorder, and epilepsy, among others. Diagnostic classifications of neurodevelopmental disorders are complicated by comorbidities among these neurodevelopmental disorders, unidentified causal genes, and growing evidence of shared genetic risk factors. </p><p>We sought to identify the genetic underpinnings of a variety of neurodevelopmental disorders, with a particular emphasis on the epilepsies, by employing next&ndash;generation sequencing to thoroughly interrogate genetic variation in the human genome/exome. First, we investigated four families presenting with a seemingly identical and previously undescribed neurodevelopmental disorder characterized by congenital microcephaly, intellectual disability, progressive cerebral atrophy, and intractable seizures. These families all exhibited an apparent autosomal recessive pattern of inheritance. Second, we investigated a heterogeneous cohort of &sim;60 undiagnosed patients, the majority of whom suffered from severe neurodevelopmental disorders with a suspected genetic etiology. Third, we investigated 264 patients with epileptic encephalopathies &mdash; severe childhood epilepsy disorders &mdash; looking specifically at infantile spasms and Lennox&ndash;Gastaut syndrome. Finally, we investigated &sim;40 large multiplex epilepsy families with complex phenotypic constellations and unclear modes of inheritance. The studied neurodevelopmental disorders exhibited a range of genetic complexity, from clear Mendelian disorders to common complex disorders, resulting in varying degrees of success in the identification of clearly causal genetic variants. </p><p>In the first project, we successfully identified the disease&ndash;causing gene. We show that recessive mutations in <italic>ASNS </italic> (encoding asparagine synthetase) are responsible for this previously undescribed neurodevelopmental disorder. We also characterized the causal mutations <italic>in vitro</italic> and studied Asns&ndash;deficient mice that mimicked aspects of the patient phenotype. This work describes ASNS deficiency as a novel neurodevelopmental disorder, identifies three distinct causal mutations in the ASNS gene, and indicates that asparagine synthesis is essential for the proper development and function of the brain.</p><p>In the second project, we exome sequenced 62 undiagnosed patients and their unaffected biological parents (trios). By analyzing all identified variants that were annotated as putatively functional and observed as a novel genotype in the probands (not observed in the unaffected parents or controls), we obtained a genetic diagnosis for 32% (20/62) of these patients. Additionally, we identify strong candidate variants in 31% (13/42) of the undiagnosed cases. We also present additional analysis methods for moving beyond traditional screens, e.g., considering only securely implicated genes, or subjecting qualifying variants from any gene to two unique analysis approaches. This work adds to the growing evidence for the utility of diagnostic exome sequencing, increases patient sizes for rare neurodevelopmental disorders (enabling more detailed analyses of the phenotypic spectrum), and proposes novel analysis approaches which will likely become beneficial as the number of sequenced undiagnosed patients grows. </p><p>In the third project, we again employ a trio&ndash;based exome sequencing design to investigate the role of <italic>de novo</italic> mutations in two classical forms of epileptic encephalopathy. We find a significant excess of <italic>de novo</italic> mutations in the &sim;4,000 genes that are the most intolerant to functional genetic variation in the human population (P = 2.9 x 10<super>&ndash;3</super>, likelihood analysis). We provide clear statistical evidence for two novel genes associated with epileptic encephalopathy &mdash; <italic>GABRB3</italic> and <italic>ALG13</italic>. Together with the 15 well&ndash;established epileptic encephalopathy genes, we statistically confirm the association of an additional ten putative epileptic encephalopathy genes. We show that only &sim;12% of epileptic encephalopathy patients in our cohort are explained by <italic>de novo</italic> mutations in one of these 24 genes, highlighting the extreme locus heterogeneity of the epileptic encephalopathies. </p><p>Finally, we investigated multiplex epilepsy families to uncover novel epilepsy susceptibility factors. Candidate variants emerging from sequencing within discovery families were further assessed by cosegregation testing, variant association testing in a case&ndash;control cohort, and gene&ndash;based resequencing in a cohort of additional multiplex epilepsy families. Despite employing multiple approaches, we did not identify any clear genetic associations with epilepsy. This work has, however, identified a set of candidates that may include real risk factors for epilepsy; the most promising of these is the <italic>MYCBP2</italic> gene. This work emphasizes the extremely high locus and allelic heterogeneity of the epilepsies and demonstrates that very large sample sizes are needed to uncover novel genetic risk factors. </p><p>Collectively, this body of work has securely implicated three novel neurodevelopmental disease genes that inform the underlying pathology of these disorders. Furthermore, in the final three studies, this work has highlighted additional candidate variants and genes that may ultimately be validated as disease&ndash;causing as sample sizes increase.</p> / Dissertation

The purpose of this essay was to investigate and compare the treatment of patients at Vilhelmsro’s hospital for children with epilepsy in Jönköping, Sweden, to the treatment of mentally deficient patients in Sweden during the 19th and 20th century. To fulfill the purpose of this investigation the administrative and medicinal archive at Jönköping’s county hospital Ryhov, and the provincial archives in Vadstena were used. The basis and the results of the investigation were drawn from studying and analyzing patient records, seizure books and enrollment charts of discharged patients, dated from 1928 to 1939. The investigation shows that although the treatment of patients at Vilhelmsro share a lot of fundamental values with the treatment of mentally deficient patients at mental institutions around Sweden at that time, such as correctness of behavior in order to fit into the norms of society, the hospital’s main concern was to educate and care for the children with epilepsy in hopes of adapting into society. This was not always possible, and the care at Vilhelmsro mainly had three outcomes which consisted of either being discharged due to recovery from illness, being discharged into the care of a more severe care facility or death from severe seizures. The hospital did concern itself with making sure that the children were molded into fitting into society’s interests and behavior and unfortunately this might have taken away from the actual medical care.

The use of traditional and natural medicines in primary healthcare or alternative therapy is on the increase. However, the safety and efficacy of these medicines have not yet been confirmed. Pharmacognosy, the study of the properties of drugs, potential drugs or drug substances of natural origin and the search for new drugs from natural resources, is therefore of extreme importance in today’s healthcare environment. Cotyledon orbiculata L. var. orbiculata (Haw.) DC., a succulent shrub that is widely distributed over the whole of southern Africa, is an example of a plant used in traditional medicine for its antiepileptic effects. Oxidative stress can either be the cause of, or be secondary to epilepsy pathogenesis. Lipid peroxidation causes the disruption of cell membranes which leads to cell destruction and, in the case of neurological disorders, neurodegeneration. Reactive species have also been found to influence neurotransmission by affecting neurotransmitter metabolism and functions. Reactive species can therefore be responsible for the development of convulsions. Conventional anti-epileptics have shown to exert neuroprotective effects but information or research regarding their ability to prevent epilepsy from becoming chronic does either not exist or is not promising. Antioxidants have potential in the treatment of epileptic seizures as well as the prevention of chronic epilepsy by preventing the effects that oxidative stress has on neurotransmitter metabolism and functions that cause alterations in neuronal excitability and seizure threshold, ultimately leading to epileptic foci. The aim of this study was to evaluate the potential of the bufadienolide orbicusides of C. orbiculata and analogues as anti-epileptic treatment through antioxidant activity. Initially the isolation of novel antioxidants from C. orbiculata leaf juice was attempted. The antioxidant activity of the concentrated juice and fractions resulting thereof were evaluated with two assays. The thiobarbituric acid (TBA) assay was used to measure the extent of lipid peroxidation and nitroblue tetrazolium (NBT) assay was used to measure superoxide scavenging activity in rat brain homogenate. The low concentrations of orbicusides prompted the determination of the activity of two commercial bufadienolides (bufalin and cinobufotalin) and two bufadienolide analogues, synthesised by the esterification of trans-androsterone and androstanolone, respectively, using coumalic acid, producing Compound 1 and Compound 2. The toxicity of the commercial bufadienolides and synthesised analogues were evaluated by using the MTT assay (a cell viability assay). C. orbiculata juice showed significant pro-oxidant activity in both assays. Bufalin showed significant pro-oxidant activity in the TBA assay. Cinobufotalin showed no significant activity. Compound 1 showed pro-oxidant activity in the TBA assay and Compound 2 showed slight antioxidant activity in the NBT assay. The commercial bufadienolides showed low cell viability, indicating significant toxicity. The synthesised analogues showed a significant reduction in toxicity (despite Compound 2 being moderately toxic) when compared to the toxicity of the commercial bufadienolides. The low concentrations of orbicusides in the plant material and the antioxidant assay results of the two commercial bufadienolides suggested that the orbicusides may not be involved in the antioxidant properties of C. orbiculata. However, the antioxidant activity of Compound 2 showed that altering the pyrone moiety of bufadienolides could possibly improve antioxidant activity. The reduced toxicity and slight antioxidant activity of the synthesised bufadienolide analogues motivates further investigation. / MSc (Pharmaceutical Chemistry), North-West University, Potchefstroom Campus, 2014

The use of traditional and natural medicines in primary healthcare or alternative therapy is on the increase. However, the safety and efficacy of these medicines have not yet been confirmed. Pharmacognosy, the study of the properties of drugs, potential drugs or drug substances of natural origin and the search for new drugs from natural resources, is therefore of extreme importance in today’s healthcare environment. Cotyledon orbiculata L. var. orbiculata (Haw.) DC., a succulent shrub that is widely distributed over the whole of southern Africa, is an example of a plant used in traditional medicine for its antiepileptic effects. Oxidative stress can either be the cause of, or be secondary to epilepsy pathogenesis. Lipid peroxidation causes the disruption of cell membranes which leads to cell destruction and, in the case of neurological disorders, neurodegeneration. Reactive species have also been found to influence neurotransmission by affecting neurotransmitter metabolism and functions. Reactive species can therefore be responsible for the development of convulsions. Conventional anti-epileptics have shown to exert neuroprotective effects but information or research regarding their ability to prevent epilepsy from becoming chronic does either not exist or is not promising. Antioxidants have potential in the treatment of epileptic seizures as well as the prevention of chronic epilepsy by preventing the effects that oxidative stress has on neurotransmitter metabolism and functions that cause alterations in neuronal excitability and seizure threshold, ultimately leading to epileptic foci. The aim of this study was to evaluate the potential of the bufadienolide orbicusides of C. orbiculata and analogues as anti-epileptic treatment through antioxidant activity. Initially the isolation of novel antioxidants from C. orbiculata leaf juice was attempted. The antioxidant activity of the concentrated juice and fractions resulting thereof were evaluated with two assays. The thiobarbituric acid (TBA) assay was used to measure the extent of lipid peroxidation and nitroblue tetrazolium (NBT) assay was used to measure superoxide scavenging activity in rat brain homogenate. The low concentrations of orbicusides prompted the determination of the activity of two commercial bufadienolides (bufalin and cinobufotalin) and two bufadienolide analogues, synthesised by the esterification of trans-androsterone and androstanolone, respectively, using coumalic acid, producing Compound 1 and Compound 2. The toxicity of the commercial bufadienolides and synthesised analogues were evaluated by using the MTT assay (a cell viability assay). C. orbiculata juice showed significant pro-oxidant activity in both assays. Bufalin showed significant pro-oxidant activity in the TBA assay. Cinobufotalin showed no significant activity. Compound 1 showed pro-oxidant activity in the TBA assay and Compound 2 showed slight antioxidant activity in the NBT assay. The commercial bufadienolides showed low cell viability, indicating significant toxicity. The synthesised analogues showed a significant reduction in toxicity (despite Compound 2 being moderately toxic) when compared to the toxicity of the commercial bufadienolides. The low concentrations of orbicusides in the plant material and the antioxidant assay results of the two commercial bufadienolides suggested that the orbicusides may not be involved in the antioxidant properties of C. orbiculata. However, the antioxidant activity of Compound 2 showed that altering the pyrone moiety of bufadienolides could possibly improve antioxidant activity. The reduced toxicity and slight antioxidant activity of the synthesised bufadienolide analogues motivates further investigation. / MSc (Pharmaceutical Chemistry), North-West University, Potchefstroom Campus, 2014

abstract: The use of bias indicators in psychological measurement has been contentious, with some researchers questioning whether they actually suppress or moderate the ability of substantive psychological indictors to discriminate (McGrath, Mitchell, Kim, & Hough, 2010). Bias indicators on the MMPI-2-RF (F-r, Fs, FBS-r, K-r, and L-r) were tested for suppression or moderation of the ability of the RC1 and NUC scales to discriminate between Epileptic Seizures (ES) and Non-epileptic Seizures (NES, a conversion disorder that is often misdiagnosed as ES). RC1 and NUC had previously been found to be the best scales on the MMPI-2-RF to differentiate between ES and NES, with optimal cut scores occurring at a cut score of 65 for RC1 (classification rate of 68%) and 85 for NUC (classification rate of 64%; Locke et al., 2010). The MMPI-2-RF was completed by 429 inpatients on the Epilepsy Monitoring Unit (EMU) at the Scottsdale Mayo Clinic Hospital, all of whom had confirmed diagnoses of ES or NES. Moderated logistic regression was used to test for moderation and logistic regression was used to test for suppression. Classification rates of RC1 and NUC were calculated at different bias level indicators to evaluate clinical utility for diagnosticians. No moderation was found. Suppression was found for F-r, Fs, K-r, and L-r with RC1, and for all variables with NUC. For F-r and Fs, the optimal RC1 and NUC cut scores increased at higher levels of bias, but tended to decrease at higher levels of K-r, L-r, and FBS-r. K-r provided the greatest suppression for RC1, as well as the greatest increases in classification rates at optimal cut scores, given different levels of bias. It was concluded that, consistent with expectations, taking account of bias indicator suppression on the MMPI-2-RF can improve discrimination of ES and NES. At higher levels of negative impression management, higher cut scores on substantive scales are needed to attain optimal discrimination, whereas at higher levels of positive impression management and FBS-r, lower cut scores are needed. Using these new cut scores resulted in modest improvements in accuracy in discrimination. These findings are consistent with prior research in showing the efficacy of bias indicators, and extend the findings to a psycho-medical context. / Dissertation/Thesis / Ph.D. Psychology 2013

This thesis explores the perspectives of people on psychogenic non-epileptic seizures (PNES). It is presented in three separate papers: a systematic literature review; an empirical research paper and a critical reflection of the research process as a whole. The systematic literature review aimed to provide a detailed understanding of stakeholder perspectives on PNES. A systematic search identified relevant studies that were subsequently synthesised using thematic analysis and the broader principles of narrative synthesis. Three broad themes relating to stakeholder perspectives were identified: the nature of PNES as a condition; diagnosis; and management and treatment issues. It was found that both patients and professionals experienced uncertainties in relation to understanding and managing the condition. This highlighted the need for further information and awareness of PNES and the development of clear treatment guidelines. Important differences in opinion were also identified between patients and professionals and consideration was given to how these may disrupt the development of effective partnerships in care. The research into patients' and families' perspectives was found to be lacking and further research was identified as being needed in this area. The empirical paper reports an exploratory qualitative study that aimed to provide an in-depth understanding of the perceived treatment needs of patients with PNES. Semi-structured interviews were conducted and findings were analysed inductively using the principles of thematic analysis. Four key themes were identified: return to normality; post-diagnostic limbo; uncertainty and apprehension about therapy; and need for validation. Patients with PNES described clear goals for their recovery and clear ideas about their treatment needs. However, following their diagnosis, many felt caught in 'limbo' due to uncertainties about their diagnosis and as a result of a lack of post-diagnostic support. Being in 'limbo' also linked to patients' uncertainties about psychology meeting their needs and for some there was apprehension about the potential negative consequences of therapy. The clinical implications of the research are discussed and recommendations for future research are made. The third paper is a critical reflection of the research process as a whole. It provides an overview and evaluation of the first two papers and personal reflections of the lead researcher are offered throughout. Implications for further research and clinical practice are offered and a summary of the research as a whole is offered.

In the last decades, although the scientific community has attempted to explain a series of complex phenomena, ranging from natural hazards to physical conditions and economic crises, aspects of their generation process still escape our full understanding. The present thesis intends to promote our understanding of the spatiotemporal behavior and the generation mechanisms that govern large and strong earthquakes, employing a broad multidisciplinary perspective for the interpretation of catastrophic events. Two main questions are debated. The first question concentrates on “whether the generation process of an extreme event has more than one facets prior to its final appearance”. In the scientific study of earthquakes, attention is drawn to the predictive capability and monitoring of different precursory observations. Among them preseismic electromagnetic emissions have been also observed indicating that the science of earthquake prediction should be from the start multidisciplinary. Drawing on recently introduced models for earthquake dynamics, that address issues such as long-range correlations, self-affinity, complexity-organization and fractal structures, the present work endeavors to further penetrate on the analysis of preseismic electromagnetic emissions and elucidate their link with the generation process of large and strong earthquakes. A second question deals with “whether there is a unified approach for the study of catastrophic events”. This question implies the possibility for common statistical behavior of diverse extreme events and the potential for transferability of methods from the study of earthquake dynamics across other fields. On these grounds, the present work extends the focus of inquiry to the analysis of electroencephalogram recordings related to epileptic seizures, in the prospect to identify common mechanisms that may explain the nature and the generation process of both phenomena, and to open up different directions for future research. Finally, with a view to consider alternative ways of studying key theoretical principles associated with the generation process of catastrophic phenomena, a relevant framework based on proposed algorithms is presented, focusing on parameters such as: the energy of earthquakes, the mean and maximum magnitude of the sample, the probability that two samples may come from the same population. Such an attempt aims to contribute to the knowledge of natural phenomena, by extending the existing theory and models and providing a few more ways for their interpretation.

Prior work in the Durst group had generated more than forty analogs of the potent anticonvulsant isoxylitone isolated isolated from a medicinal plant Delphinium denudatum Wall. The nitrile designated as TD532 was the most potent compound generated by A. Saikaley. The starting material for the synthesis of TD532 is isophorone. The observation that TD532 showed considerable potential as an anticonvulsant suggested that other cyclohexenones might have have similar activity. During this project close to fifty derivatives of cyclohex-2-enone, focusing mainly on 3-arylcylohex-2-enones, were prepared. The synthesis of these compounds is described and structure activity relationships are discussed. Based on all the available structure activity data, we have designated the indicated portion of structure A as the pharmacophore for anticonvulsant and anti-epileptic activity. The ester designated as TD561 (compound 40) showed excellent potential in both in vitro and in vivo assays. It has been shown to be a pro-drug of the corresponding acid TD562 (compound 48). These two compounds and the sodium salt of TD562 are currently undergoing final pre-clinical studies at the Center for Drug Research and Development in Vancouver. Five analogs, including TD561 are also under investigation by the Epilepsy and Seizure Division of the US National Institutes of Health.

Epilepsy affects approximately 0,5-1% of children. Epileptic seizures originate in and propagate along certain neural pathways involved in physiological processes of cognition. Consequently, cognitive impairment frequently accompanies epilepsy in childhood and contributes to diminished quality of life of these patients.The main goal of this PhD thesis was to study multiple aspects of cognitive impairment in children suffering from intractable focal epilepsy. In the first and primary study, we described for the first time the negative impact of quasi- periodic epileptiform discharges in sleep (termed "hurdles" in our work) on cognitive functions in children with focal structural epilepsy. We have also shown that epileptiform activity in sleep has a more prominent negative impact on cognitive functions than epileptiform activity in wake. Although "hurdles" are by definition generalized, they do not predict worse outcomes of epilepsy surgery, compared to controls. In the second study, we analyzed the relationship between the extent of epileptogenic zone, functional brain plasticity (evaluated by fMRI) and cognitive dysfunction in children with drug resistant temporal epilepsy. Comparing patients with isolated focal cortical dysplasia (FCD) and patients with combined pathology (FCD and hippocampal...