Moral injury (MI) is associated with severe blame-related emotion and the development of psychopathology including posttraumatic stress disorder (PTSD). Little is known about how MI events are neurally processed when PTSD is comorbid, limiting the development of tailored interventions. Thus, this thesis sought to provide a novel, multi-method examination of the biological underpinnings of moral injury and relevant behavioural correlates. Study one provides the first investigation into the neural activation patterns elicited during MI event recall in military members and public safety personnel with PTSD, relative to MI-exposed civilian controls. In PTSD, emotional processing is challenged by heightened sensory information. Here, we provide evidence of heightened viscerosensory information processing (i.e. internal gnawing or gastrointestinal constriction related to blame-based emotion) during MI event recall, which appears to exert a strong influence over cortical regions facilitating moral cognitive processes including emotion regulation, autobiographical memory integration, and social cognition. Overwhelming visceral sensations can elicit defensive behaviour including tonic immobility (TI), a defensive response that facilitates viscerosensory dampening. Interestingly, more severe negative alterations in cognition and mood were associated with viscerosensory dampening in our PTSD group, pointing towards a compensatory pattern of emotional numbing. Studies two and three explore two posttraumatic symptoms consistent with emotional numbing: alexithymia and posttraumatic TI. In study two, we explore posttraumatic TI as a survival-based dissociative response and test a new measure of posttraumatic TI. In study three, we provide evidence that alexithymia is associated with an altered (muted) pattern of emotion-specific bodily sensation. This thesis provides a framework for embodied MI event processing in PTSD and highlights the importance of assessing the somatic experience of MI and screening for TI responses and emotional numbing as part of PTSD symptomatology. The evidence presented here suggests sensorimotor-based approaches and bottom-up regulatory strategies may be useful adjuncts to MI event processing. / Dissertation / Doctor of Philosophy (PhD) / Moral injury (MI) is a psychosocial-spiritual injury that can occur when deeply held values are violated either by oneself or a trusted other; it produces considerable pain and social alienation. MI has been linked to suicide and the development of posttraumatic stress disorder (PTSD), a mental health condition associated with distressing symptoms and reduced functioning in important areas of life, including social relationships. This thesis focuses on how MI events are processed by military members and public safety personnel, who are both at risk for MI and PTSD. We investigate how shame is experienced in the brain and body, and explore how intensified visceral sensations may become overwhelming (e.g., pit in stomach, vomiting) prompting emotional numbing or difficulties remaining embodied in the present moment (e.g., zoning out, freezing up). By understanding MI event processing when PTSD is present, we hope to gain insight into more effective treatments for these individuals.

The ATP-gated P2X7 receptor (P2X7R) is a non-selective cation channel which senses high extracellular ATP concentrations and has been suggested as a target for the treatment of neuroinflammation and neurodegenerative diseases. The use of P2X7R antagonists may therefore be a viable approach for treating CNS pathologies, including epileptic disorders. Recent studies showed anticonvulsant potential of P2X7R antagonists in certain animal models. To extend this work, we tested three CNS-permeable P2X7R blocker (Brilliant Blue G, AFC-5128, JNJ-47965567) and a natural compound derivative (tanshinone IIA sulfonate) in four well-characterized animal seizure models. In the maximal electroshock seizure threshold test and the pentylenetetrazol (PTZ) seizure threshold test in mice, none of the four compounds demonstrated anticonvulsant effects when given alone. Notably, in combination with carbamazepine, both AFC-5128 and JNJ-47965567 increased the threshold in the maximal electroshock seizure test. In the PTZ-kindling model in rats, useful for testing antiepileptogenic activities, Brilliant Blue G and tanshinone exhibited a moderate retarding effect, whereas the potent P2X7R blocker AFC-5128 and JNJ-47965567 showed a significant and long-lasting delay in kindling development. In fully kindled rats, the investigated compounds revealed modest effects to reduce the mean seizure stage. Furthermore, AFC-5128- and JNJ-47965567-treated animals displayed strongly reduced Iba 1 and GFAP immunoreactivity in the hippocampal CA3 region. In summary, our results show that P2X7R antagonists possess no remarkable anticonvulsant effects in the used acute screening tests, but can attenuate chemically-induced kindling. Further studies would be of interest to support the concept that P2X7R signalling plays a crucial role in the pathogenesis of epileptic disorders.

The mammalian central nervous system maintains a delicate balance between neuronal excitation and inhibition. Conventional synaptic inhibition is mediated through the transient activity of postsynaptic γ-aminobutyric acid (GABA) at type A GABA (GABAA) receptors. A subset of GABAA receptors is also located outside of inhibitory synapses. These extrasynaptic receptors generate a tonic inhibitory conductance in response to low concentrations of extracellular GABA. Tonic inhibition broadly suppresses neuronal activity and regulates many vital processes such as sleep, consciousness and memory formation. This thesis examines the physiological effects of tonic inhibition at the cellular level and in the behaving animal. This thesis also explores whether gabapentin, a commonly used sedative, anxiolytic, and analgesic drug, enhances tonic GABAergic inhibition. I hypothesize that: (1) tonic GABAA receptor activity reduces the intrinsic excitability of neurons; (2) the activity of tonically active GABAA receptors in spinal pain pathways attenuates nociception; and (3) tonic inhibition can be upregulated by gabapentin. The results show that a tonic inhibitory current generated by α5 subunit-containing GABAA (α5GABAA) receptors reduces the excitability of hippocampal pyramidal neurons excitability by increasing the rheobase, but does not change the gain of action potential firing. A similar shunting inhibition is present in spinal cord lamina II neurons that is generated by δ subunit-containing GABAA receptors. The activity of these receptors in spinal nociceptive pathways reduces acute thermal nociception and may constrain central sensitization in a behavioural model of persistent pain. Finally, gabapentin increases a tonic inhibitory current in cultured hippocampal neurons independent from changes in the expression of α5GABAA receptors or in the concentration of GABAA receptor ligands. The results of this thesis demonstrate that tonically active GABAA receptors play an important role in the regulation of neuronal activity and nociception, and that tonic inhibition represents a novel target of therapeutic drugs.

The contractile state of vascular smooth muscle (VSM) plays a key role in blood pressure regulation. Abnormal VSM contractility characterizes hypertension and understanding the regulatory mechanisms of VSM may provide some insights to specific treatment of hypertension. Upon muscle stimulation, Ca2+, myosin light chain (MLC) phosphorylation, crossbridge cycling rates and force increase to high levels, but with time, force is maintained while all other parameters of muscle activation fall to low levels. Thus, contraction is divided temporally into early (phasic) and late (tonic) phases, as determined by the underlying regulatory mechanism. Muscles with more phasic characteristics have a higher peak phase while tonic muscles have both high peak and tonic. However, these muscles have similar contractile increases in Ca2+ despite their substantial differences in force production during the tonic phase of contraction. Myosin light chain phosphatase (MLCP) inhibition by RhoA Kinase (ROK) has been shown to contribute to this increased force production without simultaneous increases in calcium in a process known as Ca2+ sensitization.Epigastric artery (EA) has a unique regulatory mechanism controlled by the degree of stretch applied on the artery which produces an increase in tonic force maintenance. Tonic force in EA is blocked equally at all lengths by the ROK inhibitor Y-27632, suggesting ROK is the main mechanism of signal transduction activated.MLC-p is increased during basal compared to unstretched conditions for epigastric but not femoral artery (FA). Pharmacological MT depolymerization due to Colchicine incubation has been shown to release RhoGEF, a RhoA activator, and resulted in increased tonic force and MLC-p which were both inhibited by a ROK inhibitor.Additionally, KC1-stimulation appeared to activate MAPK and ROK pathways, while stretch alone activated a yet undetermined pathway, possibly ILK. KC1-induced contraction in FA activated TRP sensitive calcium channels during both peak and tonic phases. However, stretch in EA does not induce additional calcium influx. Thus, these data support the conclusion that an increase in length activates ROK and other kinases resulting in tonic force maintenance in EA.

Esta dissertação tem como objeto as vogais médias postônicas não-­finais do Português Brasileiro, especificamente a emergência das formas altas [] e [] e das médias-­baixas [] e [], estas últimas especificamente em dialetos do nordeste. Por meio do arcabouço teórico da Geometria de Traços, buscou-­se analisar os motivadores fonológicos que estão correlacionados à emergência de cada uma das formas das vogais. A principal hipótese que se buscou verificar foi se a emergência das vogais [] e [] na posição postônica não-­final, classificadas na literatura como resultantes de alçamento vocálico, está condicionada a correlatos fonológicos, tendo em vista que até então não se conseguiu encontrar um padrão que pudesse formalizar uma regra. Além disso, buscou-­se investigar se a emergência das formas médias-­baixas [, ] na postônica não-­final poderia ser formalizada por regra. Para tanto, realizou-­se um experimento de leitura de palavras em duas localidades, São Paulo e São Luís. A amostra foi constituída por 40 informantes com idades entre 20 e 30 anos, 20 de cada localidade. Os dados foram transcritos após uma verificação acústica de medição de formantes e posteriormente codificados para que testes estatísticos fossem aplicados. Com base nos resultados obtidos, observou-­se que (i) a emergência das formas altas se correlacionou à presença de uma vogal alta na átona final que compartilhasse o mesmo ponto de articulação que o da postônica não-­final, isto é, coronal para [] e labial para []; e que (ii) a emergência das médias-­baixas se correlacionou à presença da vogal dorsal, ou seja, da vogal baixa na átona final. Para a formalização de ambas as regras, faz-­se necessário assumir a proposta de Wetzels (2011) a respeito da neutralização como um mecanismo de mudança do valor do traço que garante a distinção em outro contexto. Entretanto, para que se pudesse capturar o fato de, para a emergência das vogais altas, além do traço de abertura, haver também correlação com o ponto de articulação da átona final e de que a produção das médias-­baixas é garantida pelo traço [aberto3], ambas as regras devem ser assimilatórias: do nó vocálico para as altas [, ] e do traço [+aberto3] para as médias-­baixas [, ]. / This thesis focuses on non-­final post-­tonic mid-­vowels, specifically, the emergence of the high [, ] and low-­mid vowels [, ], the latter characteristic of northeastern dialects. Based on the theoretical framework of Feature Geometry, it was intended to study the phonological motivators that are correlated with the emergence of each vowel form. The main hypothesis verified was if the emergence of [] and [] in non-­ final post-­tonic context, classified in the literature as a result of vowel rising phenomenon, is subject to phonological correlates, considering that up to this point no known study had found a pattern that could formalize a rule. In addition, it was intended to investigate whether the emergence of the low-­mid forms [, ] in non-­final post-­tonic context could also be formalized by rule. Therefore, a word reading experiment was ran in two cities, São Paulo and São Luis. The sample consisted of 40 informants, who were men and women aged between 20 and 30 years, 20 from each locality. Data were transcribed after an acoustic analysis of formant measurement and subsequently coded for statistical analysis. Based on the results, it was found that (i) the emergence of the high vowels was correlated with the presence of a high word final vowel that also shared the same place of articulation as the one in the non-­final post-­tonic context, i.e., coronal to [] and labial to []; and (ii) the emergence of mid-­low was correlated with the presence of the dorsal vowel, therefore, low vowel in word final position. To formalize both rules, it is necessary to assume Wetzels (2011) proposal that neutralization is a mechanism by which contrastive feature values are replaced by their opposite values. However, in order to capture the fact that, for the emergence of the high vowel, besides the open feature, theres also an association to the place of articulation of the word final vowel and that the production of mid-­low is guaranteed by the feature [open3], both rules should be assimilatory: the vocalic node for the high [, ] and the [+open3] feature to low-­ mid [, ].

Esta dissertação tem como objeto as vogais médias postônicas não-­finais do Português Brasileiro, especificamente a emergência das formas altas [] e [] e das médias-­baixas [] e [], estas últimas especificamente em dialetos do nordeste. Por meio do arcabouço teórico da Geometria de Traços, buscou-­se analisar os motivadores fonológicos que estão correlacionados à emergência de cada uma das formas das vogais. A principal hipótese que se buscou verificar foi se a emergência das vogais [] e [] na posição postônica não-­final, classificadas na literatura como resultantes de alçamento vocálico, está condicionada a correlatos fonológicos, tendo em vista que até então não se conseguiu encontrar um padrão que pudesse formalizar uma regra. Além disso, buscou-­se investigar se a emergência das formas médias-­baixas [, ] na postônica não-­final poderia ser formalizada por regra. Para tanto, realizou-­se um experimento de leitura de palavras em duas localidades, São Paulo e São Luís. A amostra foi constituída por 40 informantes com idades entre 20 e 30 anos, 20 de cada localidade. Os dados foram transcritos após uma verificação acústica de medição de formantes e posteriormente codificados para que testes estatísticos fossem aplicados. Com base nos resultados obtidos, observou-­se que (i) a emergência das formas altas se correlacionou à presença de uma vogal alta na átona final que compartilhasse o mesmo ponto de articulação que o da postônica não-­final, isto é, coronal para [] e labial para []; e que (ii) a emergência das médias-­baixas se correlacionou à presença da vogal dorsal, ou seja, da vogal baixa na átona final. Para a formalização de ambas as regras, faz-­se necessário assumir a proposta de Wetzels (2011) a respeito da neutralização como um mecanismo de mudança do valor do traço que garante a distinção em outro contexto. Entretanto, para que se pudesse capturar o fato de, para a emergência das vogais altas, além do traço de abertura, haver também correlação com o ponto de articulação da átona final e de que a produção das médias-­baixas é garantida pelo traço [aberto3], ambas as regras devem ser assimilatórias: do nó vocálico para as altas [, ] e do traço [+aberto3] para as médias-­baixas [, ]. / This thesis focuses on non-­final post-­tonic mid-­vowels, specifically, the emergence of the high [, ] and low-­mid vowels [, ], the latter characteristic of northeastern dialects. Based on the theoretical framework of Feature Geometry, it was intended to study the phonological motivators that are correlated with the emergence of each vowel form. The main hypothesis verified was if the emergence of [] and [] in non-­ final post-­tonic context, classified in the literature as a result of vowel rising phenomenon, is subject to phonological correlates, considering that up to this point no known study had found a pattern that could formalize a rule. In addition, it was intended to investigate whether the emergence of the low-­mid forms [, ] in non-­final post-­tonic context could also be formalized by rule. Therefore, a word reading experiment was ran in two cities, São Paulo and São Luis. The sample consisted of 40 informants, who were men and women aged between 20 and 30 years, 20 from each locality. Data were transcribed after an acoustic analysis of formant measurement and subsequently coded for statistical analysis. Based on the results, it was found that (i) the emergence of the high vowels was correlated with the presence of a high word final vowel that also shared the same place of articulation as the one in the non-­final post-­tonic context, i.e., coronal to [] and labial to []; and (ii) the emergence of mid-­low was correlated with the presence of the dorsal vowel, therefore, low vowel in word final position. To formalize both rules, it is necessary to assume Wetzels (2011) proposal that neutralization is a mechanism by which contrastive feature values are replaced by their opposite values. However, in order to capture the fact that, for the emergence of the high vowel, besides the open feature, theres also an association to the place of articulation of the word final vowel and that the production of mid-­low is guaranteed by the feature [open3], both rules should be assimilatory: the vocalic node for the high [, ] and the [+open3] feature to low-­ mid [, ].

In the Romantic period, expanded tonality offers a creative challenge to composers as they explore new ways of establishing the hierarchy of pitches and utilizing the chromatic and diatonic resources. Prominent compositional techniques of this period include the use of linear harmony with less clearly defined root movements, the structural placement of dominant function, new approaches that redefine tonal stability, motivic treatment that generates harmony and form, flexible treatment of rhythm and meter, and functional treatment of chromatic pitches. This study explores the ways in which characteristics of the Romantic period are influenced by the upper/lower leading tone and the effects of compositional treatment on the expansion of tonality. In addition, this study includes two supportive concepts: (1) the wedge and toggle switch by David Witten and (2) The Neapolitan Complex by Christopher Wintle. In describing techniques in expanded tonality, excerpts from compositions by Robert Schumann and Johannes Brahms (both prominent composers of the Romantic period) are used to establish the significance of these techniques. In the fourth movement of Sonata in E minor "Undine," by Carl Reinecke, the structural treatment of the upper/lower leading tones to tonic and dominant are very prominent features that contribute significantly to the development of the concepts in this study.

Tonic immobility, TI, is an innate fear response in many vertebrate animals, induced by brief physical restraint. It is a widespread theory that chickens (Gallus gallus) reared under stressful conditions react stronger in tonic immobility tests, i.e. stay still and lay still for a longer period of time, than individuals reared under more stress-free conditions. In our study we attempted to see if stressful conditions early in life (temporary isolation from the flock on a daily basis) had any effect on how the individuals handled the stressful and fear-evoking experience of tonic immobility tests. A total of 77 chickens of the HyLine strain of White Leghorn were used. Three sets of tests were performed; first at the hatchery facility at Linköpings universitet; second after a stressful experience – the moving to the Wood-Gush facility at Vreta jordbruksgymnasium; and a random sample with about half of the birds the third time, also at Vreta. The results were inconclusive, but pointed more in the direction of the early-in-life stress having no effect on TI tests rather than the other way around.

Animals can show different responses to fear for example by playing dead when there is no possibility to escape. This response is called tonic immobility (TI) and is a well-established test of fear to evaluate fearfulness. Long durations of TI are generally considered as high levels of fearfulness. Physiological changes observed during tonic immobility suggest that there are changes in the autonomic nervous system (ANS) strongly involved in this process. The main objective for this study was to analyse duration of tonic immobility and heart and ventilation rate during tonic immobility in three different species; domesticated High Andean chickens (Gallus gallus), wild-caught Ornate Tinamous (Nothoprocta ornata) and Chilean Tinamous born in captivity (Nothoprocta perdicaria). In this study needle electrodes were used to measure heart and ventilation rate. The time following induction of tonic immobility (i.e. after holding the bird on its back for 15 s) was characterized by a large increase in heart and ventilation rate. During tonic immobility a progressive decrease in heart and ventilation rate was observed in all species, significant in all cases except for heart rate between start and end of TI in chickens. The duration of TI was significantly longer in Ornate Tinamou compared to Chilean Tinamou and chickens. The same was observed in latency to first head movement. TI is probably controlled by the autonomic nervous system, but a heart rate variability analysis has to be done in order to determine the different relative contributions of the sympathetic and parasympathetic systems in these species.

The purpose of this study was to design and test five SNP markers in an inbred chicken cross between Red Junglefowl and domestic White Leghorn of the 8th generation. The markers lie in a region affecting the tonic immobility behaviour which differs significantly between the two species. The markers could be identified by usage of PCR and pyrosequencing. The data obtained were further used in a small scale quantitative trait locus (QTL) analysis. QTL analysis is a statistical method to link phenotypic traits to genotypic data. Four out of five markers could be genotypes and thereby, made it possible to proceed with the QTL analysis. The results showed that there is no QTL associated with the markers identified. The two flanking markers were closest to a significant difference between genotypes and it is therefore a possibility that a QTL lies close further down or up the searched region. From the line map it is indicated that there is little recombination in the marker region.