Delivery of neurotrophins to the injury site via cellular transplants or viral vectors administration has previously been shown to promote recovery of locomotor behavior in the absence of locomotor training in adult spinalized animals. Viral vectors still pose clinical concerns associated to recombinant genetics and the lack of understanding of how they react with the human immune system. Delivery via graft of autologous fibroblast engineered to produce brain derived neurotrophic factor (BDNF) and Neurotrophin-3 (NT-3) has been shown as a valuable method; however, the need for multiple invasive surgeries, along with the impossibility of delivering a controlled and constant dosage of protein are serious obstacles to obtaining approval by the FDA. The present study was aimed at evaluating the efficacy of BDNF delivered to the lumbar locomotor centers using a clinically translational delivery method at restoring stepping abilities in a large animal model of spinal cord injury. We wanted to evaluate if intrathecal delivery of BDNF to the lumbar spinal cord would promote a locomotor recovery as effective as delivery to the injury site, even at doses low enough not to trigger the side effects observed at high doses. A programmable and implantable mini-pump was used to intrathecally deliver a 50 ng/day dose of BDNF to the lumbar spinal cord for 35 days after spinal thoracic transection. Kinematic evaluation was conducted before, 3 and 5 weeks after injury/pump implant. Ground reaction forces (GRFs) analysis was performed 5 weeks after injury to evaluate the animals’ ability to weight support during locomotion and standing trials. Results showed that treated cats were capable of executing weight-bearing plantar stepping at all velocities tested (0.3-0.8 m/s). Control cats did not recover stepping ability, especially at higher velocities, and dragged their hind paws on the treadmill. We were also interested in measuring the extent of BDNF diffusion within the lumbar area of the spinal cord and the potential damage to the cord caused by catheter insertion. Immunohistological evaluation showed higher BDNF expression in the dorsal root ganglions, with BDNF Immuno-Histo Chemistry (IHC) extending from L3 to L7 in all treated cats. BDNF was also found within multiple cells of the grey matter, although the levels were not significantly higher than background density. Glial fibrillary acidic protein (GFAP) stain was used to measure the immunohistological reaction of the spinal cord to the implanted catheter, and to establish the safety of the delivery method. Gross examination of the spinal cord post-mortem revealed no damage to the cord or the roots with minimal encapsulation of the catheter/pump. Minimal tissue inflammation was revealed by the GFAP stain, underlying the safety of our method. We also wanted to investigate and characterize changes in the locomotor circuitry induced by BDNF delivery. Comparison of multiunit activity in the lumbar area between BDNF treated and non-treated cats allows a better understanding of the mechanism of action of BDNF on the spinal interneurons. This was accomplished by extracellularly recording lumbar interneuronal firing during air-stepping in a 5 weeks post-injury terminal experiment. The cord was exposed at the lumbar level between the L3 and L7 spinal segments. In-vivo recordings of spinal extracellular signals were conducted using two 64 channels microelectrode arrays inserted at the dorsal root entry zone to depths of ~3000µm and ~1500µm. The ability to record simultaneous activity of multiple single neurons made it possible to study the extent to which spiking activity in a given neuron is related to concurrent ensemble spiking activity. A point process generalized linear model (PP-GLM) approach was used to assess the strength of the connections between spike trains. Interneurons activity was assessed in terms of average firing rate, signal-to-noise ratio (SNR), and number of active units per trial. Although BDNF infusion in the lumbar segments did not show significant effect on strengthening synaptic connections, we did find greater multiunit activity in the treated animals, sign of a potential BDNF-induced increase in interneuronal activation, which could be likely involved in recovery of stepping ability after SCI. Together, findings from these aims demonstrated the therapeutic potential of intrathecal lumbar BDNF delivery in spinalized animals. Constant infusion of BDNF to the locomotor centers promotes locomotor recovery similar to training or delivery to the injury site via cellular transplants after complete SCI. Intrathecal delivery by an implantable/programmable pump is a safe and effective method for delivery of a controlled BDNF dosage; it poses minimal risks to the cord and is clinically usable. Lastly, this study confirmed the major involvement of BDNF in increasing the activity of the interneurons in the locomotor circuitry, opening the door to further investigating the mechanism through which neurotrophins induce recovery of locomotion. / Bioengineering

Dissertation (PhD)--Stellenbosch University, 2001. / ENGLISH ABSTRACT: Obsessive-compulsive disorder (OCD) is characterized by intrusive thoughts (obsessions) and repetiti ve mental acts or behaviours (compulsions) . For many years, it was considered a rather uncommon condition, caused by unconscious conflict, and somewhat resistant to treatment. In recent decades, however, it has emerged that OCD is a highly prevalent disorder, mediated by particular neuroanatomical circuits (e.g. striatal pathways) and neurochemical systems (e.g. the serotonin system), and responsive to treatment with serotonin reuptake inhibitors (SRIs) . Nevertheless, many questions remain; about the specificity of neuroanatomical findings to OCD, about the role of the multiple serotonin (5-HT) receptor subtypes (e.g. 5-HT10)' and about the appropriate pharmacotherapy for patients resistent to SRI treatment? In a series of studies, 1) the neuroanatomy of OCD was assessed by means of magnetic resonance imaging and neuropsychological testing, 2) the neurochemistry of OCD was assessed by means of functional brain imaging after administration of a 5-HT10 agonist, and 3) the pharmacotherapy of OCD was explored in a series of treatment-refractory OCD and OCD spectrum disorder patients using SRI augmentation with a dopamine blocker. Although no significant difference was found in the volume of the caudate in women with OCD and controls, there was a significant correlation between caudate volume and neuropsychological dysfunction in patients, consistent with evidence of striatal involvement in OCD. Functional imaging demonstrated behavioural heterogeneity, but brain-behaviour correlations were positive, consistent with preclinical evidence of a role for the 5-HTlD receptor in the mediation of OCD. Finally, preliminary treatment findings with dopamine blocker augmentation of a SRI were promising, consistent with preclinical understandings of the interactions between the dopamine and serotonin systems. Although oeD is a complex disorder, a number of future research avenues hold promise for providing a thorough delineation of its pathogenesis. / AFRIKAANSE OPSOMMING: Obsessief-kompulsiewe steuring (OKS) word gekenmerk deur indringende gedagtes (obsessies) en herhalende gedagtes of gedrag (kompulsies). Vir baie jare is dit beskou as 'n redelik seldsame toestand wat veroorsaak word deur onbewustelike konflik, en wat in 'n mate teen behandeling weerstandig is. Meer onlangs het dit egter na vore getree as 'n toestand wat baie dikwels voorkom, wat deur spesifieke neuroanatomiese siklusse (bv. striatale bane) en neurochemiese sisteme (bv. die serotonien-sisteem) teweeg gebring word, en wat op behandeling met serotonien heropname inhibeerders (SHIs) reageer. Nogtans is daar steeds baie vrae; oor die spesifisiteit van neuroanatomiese bevindinge vir OKS, oor die rol van die veelvuldige serotonien (5-HT) reseptor subtipes (bv. 5- HT1D), en oor die toepaslike farmakoterapie vir pasiënte wat weerstandig is vir SHI behandeling. In' n reeks van navorsingstudies, is 1.) die neuroanatomie van OKS deur middel van magnetiese resonans beelding en neurosielkundige toetse ondersoek, 2. ) die neurochemie van OKS deur middel van funksionele breinbeelding na toediening van 'n 5-HT1D agonis bepaal, en 3.) die farmakoterapie van OKS in 'n reeks van behandelingsweerstandige OKS en OKS-spektrum steuring pasiënte - waar gebruik gemaak is van SHI aanvulling met 'n dopamien-blokker - ondersoek. Alhoewel daar geen beduidende verskil in die volume van die caudata in vroue met OKS en kontroles gevind is nie, was daar 'n beduidende korrelasie tussen die caudata volume en neurosielkundige wanfunksionering in pasiënte, in ooreenstemming met striatale betrokkenheid in OKS. Funksionele beelding het positief, in demonstreer, maar ooreenstemming met brein-gedrag pre-kliniese heterogeneïteit korrelasies was in gedrag bewyse vir 'n rol vir die 5-HT1D reseptor in die bemiddeling van OKS. Ten laaste, voorlopige behandelingsbevindinge oor dopamienblokker aanvulling van 'n SHI is belowend, in ooreenstemming met v

<p>УВОД: Епилепсију карактерише појава рекурентних непровоцираних напада који су манифестација појачане активности појединих епилептички измењених неуронских група у мозгу. Електроенцефалографија (EEG), је једина метода која може открити присуство фокалних или генерализованих епилептиформних пражњења, и пружити податке који су неопходни за синдромску класификацију епилепсија. МАТЕРИЈАЛ И МЕТОДЕ: Истраживање је обухватило 229 oсоба након првог епилептичког напада. Све особе су испитиване према дијагностичком протоколу који је обухватио анамнестичке и клиничке податке као и резултате допунских EEG и неурорадиолошких (CT, MRI) испитивања. Патолошки EEG налаз је описан као епилептиформан или неепилептиформан, генерализованог, нелатерализованог или фокалног типа. Неурорадиолошки снимак је сматран патолошким уколико је откривена потенцијално епилептогена лезија. Класификација типа и етиологије првог епилептичког напада је извршена у односу на важеће критеријуме. РЕЗУЛТАТИ: Нападе непознатог узрока је имало 51% особа, док је 49% особа имало симптоматске нападе. Најчешћи су били напади генерализованог типа (примарни или секундарни). Патолошки измењен EEG је регистрован код 76% особа, 47% је имало регистровану интерикталну епилептиформну активност (28% фокалну, 16% генерализовану), а&nbsp; у 31% EEG регистрација је бележена спора интериктална активност. Након депривације спавања, епилептиформна активност која је регистрована на EEG-у код 60% особа, била је најчешће фокалног типа (37%). Фактори који су дали статистички значајан допринос појави интерикталне епилептиформне активности су били старост (р 0,000; OR 1,036; 95%CI 1,021-1,051) и тип првог напада (р 0,016; OR 1,00a - 1,937; 95%CI 1,130-3,319). Ризик од рекурентних напада је био највиши код жаришних лезија (р=0,012) које су смештене у фронталном или темпоралном региону (р=0,012).&nbsp; ЗАКЉУЧАК: У периоду након првог непровоцираног напада епилептиформна активност је била чешћа код особа млађе старостне доби са генерализованим типом напада у односу на старије особе са жаришним типом напада. Иако EEG може помоћи у сагледавању епилептичке природе кризе свести, посебно код млађих особа, ипак код многих, дијагноза епилепсије зависи од појаве поновног епилептичког напада.</p> / <p>UVOD: Epilepsiju karakteriše pojava rekurentnih neprovociranih napada koji su manifestacija pojačane aktivnosti pojedinih epileptički izmenjenih neuronskih grupa u mozgu. Elektroencefalografija (EEG), je jedina metoda koja može otkriti prisustvo fokalnih ili generalizovanih epileptiformnih pražnjenja, i pružiti podatke koji su neophodni za sindromsku klasifikaciju epilepsija. MATERIJAL I METODE: Istraživanje je obuhvatilo 229 osoba nakon prvog epileptičkog napada. Sve osobe su ispitivane prema dijagnostičkom protokolu koji je obuhvatio anamnestičke i kliničke podatke kao i rezultate dopunskih EEG i neuroradioloških (CT, MRI) ispitivanja. Patološki EEG nalaz je opisan kao epileptiforman ili neepileptiforman, generalizovanog, nelateralizovanog ili fokalnog tipa. Neuroradiološki snimak je smatran patološkim ukoliko je otkrivena potencijalno epileptogena lezija. Klasifikacija tipa i etiologije prvog epileptičkog napada je izvršena u odnosu na važeće kriterijume. REZULTATI: Napade nepoznatog uzroka je imalo 51% osoba, dok je 49% osoba imalo simptomatske napade. Najčešći su bili napadi generalizovanog tipa (primarni ili sekundarni). Patološki izmenjen EEG je registrovan kod 76% osoba, 47% je imalo registrovanu interiktalnu epileptiformnu aktivnost (28% fokalnu, 16% generalizovanu), a&nbsp; u 31% EEG registracija je beležena spora interiktalna aktivnost. Nakon deprivacije spavanja, epileptiformna aktivnost koja je registrovana na EEG-u kod 60% osoba, bila je najčešće fokalnog tipa (37%). Faktori koji su dali statistički značajan doprinos pojavi interiktalne epileptiformne aktivnosti su bili starost (r 0,000; OR 1,036; 95%CI 1,021-1,051) i tip prvog napada (r 0,016; OR 1,00a - 1,937; 95%CI 1,130-3,319). Rizik od rekurentnih napada je bio najviši kod žarišnih lezija (r=0,012) koje su smeštene u frontalnom ili temporalnom regionu (r=0,012).&nbsp; ZAKLJUČAK: U periodu nakon prvog neprovociranog napada epileptiformna aktivnost je bila češća kod osoba mlađe starostne dobi sa generalizovanim tipom napada u odnosu na starije osobe sa žarišnim tipom napada. Iako EEG može pomoći u sagledavanju epileptičke prirode krize svesti, posebno kod mlađih osoba, ipak kod mnogih, dijagnoza epilepsije zavisi od pojave ponovnog epileptičkog napada.</p> / <p>INTRODUCTION: Epilepsy is characterized by recurrent, unprovoked seizures defined as the manifestation of epileptic excessive activity of neurons in the brain. Electroencephalography (EEG) provides a different focal and generalized epileptiform discharges and essential information for the syndromic classification of epilepsy. MATERIALS AND METHODS: The investigation included 229 patients with first epileptic seizure. Assesment of the first seizure presentation requires systematic clinical approach and diagnostic tests such as EEG and neuroradiological methods. In all the patients first time EEG had been performed within a month of the event. The presence of abnormal EEG activity was classified as epileptic or non-epileptic, of focal, nonlateralized or generalized type. RESULTS: 51% of all persons had a presumed seizure of unknown origin and 49% had presumed remote symptomatic seizures. About 73% had generalized seizures (primarily or secondarily). Abnormal single EEGs were present in 76%, with epileptiform activity in 47% (28% focal, 16% generalized), and nonepileptic slowing activity in 31%. Epileptiform EEG activity was present in 60%, and was predominantely focal (37%) if registration was done after sleep-deprivation. Statistical significance on interictal epileptorm activity was registered in younger age of first seizure oncet (р 0,000; OR 1,036; 95%CI 1,021-1,051) and with generalized seizures (р 0,016; OR 1,00a - 1,937; 95%CI 1,130-3,319). Reccurence risk increased in presence of focal type first seizure (p=0,012), and in frontal or temporal localization of epileptogenic lesions (p=0,012). CONCLUSION: Following a single unprovoked presumed seizure epileptiform activity was most common in younger patients with generalized seizures compared with older individuals with focal seizures. Reccurence risk increased in focal frontal and temporal seizures. Although the EEG is particularly helpful in supporting the epileptic nature of the event in younger patients, still many patients need further seizure reccurence, to be diagnosed.</p>

Notre travail a porté : 1) sur l'étude des modifications d'excitabilité corticale au cours du traitement des poussées de sclérose en plaques (SEP) et de l'évolution naturelle des formes progressives de SEP ; 2) sur la caractérisation du tremblement d'action, fréquemment observé dans le cadre de cette maladie et qui constitue une importante source de handicap. Ceci a conduit à la réalisation de 5 études. La première étude a démontré que l'amélioration rapide des performances motrices observées à la suite du traitement de poussées de SEP par une corticothérapie administrée en flash quotidien sur plusieurs jours, s'accompagnait de modifications significatives d'excitabilité corticale étudiée par stimulation magnétique transcrânienne. Ces modifications portaient sur la balance d'influences GABAergiques et glutamatergiques au sein du cortex moteur. Cette augmentation d'excitabilité survient bien avant toute possibilité de remyélinisation ou de régénérescence axonale et constitue donc une amélioration fonctionnelle induite par le traitement. Dans la deuxième étude, différents paramètres d'excitabilité corticale ont été suivis sur un an chez des patients présentant une forme progressive de SEP, traitée ou non traitée. Cette étude a mis en évidence une diminution de l'inhibition intracorticale et une augmentation du seuil moteur au repos chez les patients non traités, accompagnant une augmentation des scores cliniques de handicap. En revanche, les patients traités restaient stables, aussi bien sur le plan neurophysiologique que clinique. Ces résultats montrent que l'évolution de la SEP progressive est associée à une altération globale et évolutive de l'excitabilité du cortex moteur, portant aussi bien sur les neurones pyramidaux que sur les circuits de contrôle inhibiteur, probablement liée à l'aggravation de la perte neuronale au fil du temps. Nos résultats montrent également que différents types de traitement immunomodulateur peuvent arrêter ce cours évolutif. Dans une troisième étude, nous avons caractérisé l'existence d'un tremblement chez 32 patients atteints de SEP au moyen d'enregistrements électromyographiques et accélérométriques réalisés au membre supérieur. Ces enregistrements n'ont permis de confirmer l'existence d'un véritable tremblement que chez un seul patient. L'étude neurophysiologique concomitante de circuits de contrôle cérébelleux et protubérantiels a permis de montrer que la plupart des aspects cliniques de tremblement dans la SEP ne révélait en fait qu'un pseudo-tremblement lié en grande partie à des dysfonctions cérébelleuses. Nos deux dernières études ont permis de préciser ce résultat, au moyen d'une analyse du signal électromyographique et accélérométrique par décomposition modale empirique associée à une transformée de Hilbert-Huang. Cette méthode d'analyse apparaît valide et performante pour caractériser les tremblements et pseudo-tremblements survenant dans la SEP et les distinguer d'autres types de tremblement, comme le tremblement essentiel. / Our work focused on: 1) the study of cortical excitability changes in the treatment of multiple sclerosis (MS) relapses and the natural history of progressive forms of MS; 2) the characterization of action tremor, which is frequently observed in the course of the disease and is a major source of disability. This has led to the realization of five studies. The first study demonstrated that a rapid improvement in motor performance can be observed following treatment of MS relapses by intravenous corticosteroids administered daily over several days, accompanied by significant changes in cortical excitability parameters studied by transcranial magnetic stimulation techniques. These changes focused on the balance between GABAergic and glutamatergic influences in the motor cortex. An increase in cortical excitability occurs well before any possibility of remyelination or axonal regeneration, demonstrating a functional improvement induced by the treatment. In the second study, different parameters of cortical excitability were followed over one year in patients with progressive MS, treated or untreated. This study showed a decrease of intracortical inhibition and increased motor threshold at rest in untreated patients, accompanying a worsening of clinical disability scores. In contrast, treated patients remained stable, both on clinical and neurophysiological parameters. These results show that the evolution of progressive MS is associated with a global and progressive impairment of motor cortex excitability, concerning both pyramidal neurons on inhibitory control circuits, probably due to the progression of cortical neuronal loss over time. Our results also showed that different types of immunomodulatory therapy can stop this evolutionary course. In a third study, we characterized the existence of action tremor in 32 MS patients using electromyographic and accelerometer recordings in the upper limb. These recordings confirmed the existence of a real tremor in only one patient. Concomitant neurophysiological study of cerebellar and pontine circuits of control showed that most of the clinical aspects of tremor in MS in fact revealed a pseudo-tremor largely due to cerebellar dysfunctions. Our last two studies have clarified this result by means of an analysis of the electromyographic and accelerometer signal using empirical mode decomposition associated with Hilbert-Huang transform. This method of analysis appears valid and effective to characterize tremor or pseudo-tremor occurring in MS patients and to distinguish them from other types of tremor, such as essential tremor.

The neural processes used to extract perceptual features of vocal calls, and subsequently to re-integrate those features to form a coherent auditory object, are poorly understood. In this thesis, extracellular recordings were carried out in order to investigate how the temporal envelope, pitch, timbre and spatial location of communication sounds are represented by neurons in two core and three belt areas of ferret (Mustela putorius furo) auditory cortex. Potential neural underpinnings of auditory perception were tested using neurometric analysis to relate the reliability of neural responses to the performance of ferret and human listeners on psychophysical tasks. I found that human listeners' discrimination of the temporal envelopes of vocalization sounds matched the best neurometrics calculated from the temporal spiking patterns of ferret cortical neurons. Neurometric scores based on the spike rates of cortical neurons accounted for ferrets' discrimination of the pitch of artificial vowels. I show that most auditory cortical neurons are modulated by a number of stimulus features, rather than being tuned to only one feature. Neurons in the core auditory cortical fields often respond uniquely to particular combinations of pitch and timbre features, while those in belt regions respond more linearly to feature combinations. Subtle differences in the sensitivity of neurons to pitch, timbre and azimuthal cues were found across cortical areas and depths. These results suggest that auditory cortical neurons provide widely distributed representations of vocalizations, and a single neuron can often use combinations of spike rate and temporal spiking responses to encode multiple sound features.

Um zu verstehen, wie das Gehirn von Säugetieren funktioniert, untersuchen wir wie neuronale Aktivität einerseits zu Kognition beträgt und andererseits komplexe Verhaltensweisen ermöglicht. Im Fokus dieser Doktorarbeit stehen dabei zwei Regionen der Großhirnrinde der Ratte: der parahippocampale Cortex und der motorische Cortex. Im ersten Teil haben wir neuronale Schaltkreise im parahippocampalen Cortex und in den oberen Schichten des enthorhinalen Cortex untersucht, während Ratten ihre Umgebung räumlich erkunden. Diese beiden Regionen tragen wesentlich zum Orientierungssinn bei. Dabei haben wir herausgefunden, dass anatomische Identität und Einbindung in den Microschaltkreis einerseits räumliche neuronale Signale, wie zum Beispiel der Aktivität von grid cells, border cells und head-direction cells, bestimmen. Andererseits tragen diese beiden Eigenschaften auch zur temporalen Präzision neuronaler Signale bei, wie zum Beispiel in Form von spike bursts, theta Modulation und phase precession. Im zweiten Teil dieser Doktorarbeit untersuchen wir die Aktivität von Neuronen im Vibrissen Motorcortex während komplexer Bewegungsabläufe der Schnurrhaare, die dem natürlichen Repertoire der Ratte entstammen: eigeninitiierte Bewegungen in freier Luft, Berührung von Artgenossen zur sozialen Interaktion und das Abtasten von Objekten. Dabei haben wir herausgefunden, dass neuronale Aktivität im Motorcortex während der Bewegung der Schnurrhaare unterdrückt ist, dass elektrische Microstimulation zum Rückzug der Schnurrhaare führt und, dass pharmakologische Blockade Bewegung der Schnurrhaare fördert. Um diese überraschende Beobachtung in einen breiteren Kontext zu integrieren, endet dieser Teil mit einer Bewertung der Literatur zu der bewegungsunterdrückenden Wirkung von Motorcortex Aktivität bei Nagetieren, Primaten und Menschen. / In order to understand how the mammalian brain works, we must investigate how neural activity contributes to cognition and generates complex behavioral output. In this thesis I present work, which focuses on two regions of the cerebral cortex of rats: parahippocampal cortex and motor cortex. In the first part of the thesis we investigate neural circuits in the parasubiculum and the superficial medial enthorhinal cortex, two structures that play a key role in spatial cognition. Briefly, we find that the in these regions, anatomical identity and microcircuit embedding is a major determinant of both spatial discharge patterns (such as the discharge patterns of grid cells, border cells and head-direction cells) and temporal coding features (such as spike bursts, theta-modulation and phase precession). In the second part of the thesis we investigate the activity of neurons in vibrissa motor cortex during complex motor behaviors, which play a vital role in rat ecology: self-initiated bouts of exploratory whisking in air, whisking to touch conspecifics during social interactions and whisking to palpate objects. Briefly, we find that neural activity decreases during whisking behaviors, that microstimulation leads to whisker retraction and that pharmacological blockade increases whisker movement. Thus, our observations collectively suggest that a primary role of vibrissa motor cortex activity is to suppress whisking behaviors. The second part of the thesis concludes with a literature review of motor suppressive effects of motor cortical activity across rodents, primates and humans to put this unexpected finding in a broader context.

Eine wesentliche Aufgabe auditorischer Systeme besteht in der Erkennung und Klassifikation verhaltensrelevanter Signale. Die akustischen Kommunikationssignale vieler Feldheuschrecken zeichnen sich durch artspezifische Modulationen der Signalamplitude aus, die im Kontext der Partnerwahl zur Erkennung der eigenen Art genutzt werden. Die Kommunikation ist jedoch auch als Basis für sexuelle Selektion von Interesse - einer Abschätzung der Qualität des Senders anhand der akustischen Signale, welche eine Bewertung subtiler Variationen der artspezifischen Musters erfordert. Das Ziel dieser Arbeit bestand darin zu untersuchen, wie amplitudenmodulierte akustische Signale in den Antworten identifizierter auditorischer Interneurone der zweiten und dritten Verarbeitungsstufe repräsentiert werden, insbesondere, wie gut sie anhand dieser Antworten unterscheidbar sind. Dazu wurden (i) sinusförmig amplitudenmodulierte Stimuli genutzt und die Parameter Modulationsfrequenz und Modulationstiefe systematisch variiert, (ii) individuelle Gesänge der gleichen Art, und (iii) im Grundmuster zeitlich reskalierte Gesänge. Lokale Interneurone zeichneten sich aus durch: ein oft sehr hohes zeitliches Auflösungsvermögen, hohe Empfindlichkeit gegenüber Schwankungen der Signalamplitude, sowie gute Unterscheidbarkeit der sinusförmig amplitudenmodulierten Signale und der Gesänge auf der Basis von Spikeantworten. Bei den aufsteigenden Interneuronen nahm die Fähigkeit zur zeitlichen Ankopplung der Spikes an die Amplitudenmodulationen der Stimuli ab, was sich auch in deren reduzierter Unterscheidbarkeit äußerte. Ursächlich hierfür war einerseits die Zunahme der Antwortvariabilität (Jitter der Spikezeitpunkte), aber auch verstärkt auftretende Filtereigenschaften. Auf dieser dritten Verarbeitungsebene kommt es zu einer stärkeren Spezialisierung auf bestimmte zeitliche Aspekte des Stimulus, die als Grundlage einer verhaltensrelevanten Klassifikation von akustischen Signalen interpretiert werden kann. / A central task of auditory systems is the recognition and classification of behaviorally relevant signals. The communication signals of many grasshoppers can be characterized by a species-specific pattern of amplitude modulation, which is mainly used for species recognition in the context of mate finding. Additionally, the communication is also of interest with respect to sexual selection - an evaluation of the signaler''s quality from the signal pattern, which requires the quantification of subtle variations of the common species-specific pattern.The goal of this study was to investigate how amplitude modulated acoustic signals are represented in the responses of identified 2nd and 3rd order auditory interneurons, particularly, how well they can be discriminated on the basis of the responses. For this (i) sinusoidal amplitude modulated stimuli were used and the parameters modulation frequency and modulation depth were systematically varied, (ii) individual songs of the same species and (iii) songs with temporal rescaled basic pattern were presented. Local interneurons can be characterized by: mostly high temporal resolution capacities, high sensitivity to fluctuations of the signal amplitude as well as a good distinguishability of sinusoidal amplitude modulated stimuli and songs on the basis of the spike trains. In ascending interneurons the synchronization to the amplitude modulations decreased, which also appeared in a reduced discrimination performance. This is caused by an increase of response variability (jitter of spike timing) but also by distinctive filter properties of the respective neurons. Neurons on this third processing level exhibit a greater specialization to particular temporal aspects of the stimulus. This can be interpret as a basis of a behaviorally relevant classification of acoustic signals.

INTRODUÇÃO: A técnica de eletromiografia de fibra única (EMGFU), mediante análise do jitter, é o método neurofisiológico mais sensível para a confirmação do distúrbio da junção neuromuscular na miastenia gravis (MG). Os registros são tradicionalmente obtidos com agulha de fibra única, de alto custo e reutilizável. Por causa da necessidade atual do uso de material descartável, a agulha concêntrica vem sendo utilizada em substituição à agulha de fibra única. A técnica utilizada é semelhante, porém os potencias de ação para a análise do jitter são obtidos com eletrodo de agulha concêntrica (Eletromiografia de fibra única - jitter com agulha concêntrica, EMGFU-JAC). Contudo, os estudos são escassos e as metodologias utilizadas são heterogêneas com a utilização dessa agulha. OBJETIVOS: Este estudo tem por objetivo mensurar os valores de jitter obtidos com agulha concêntrica, no músculo Orbicularis Oculi, em sujeitos saudáveis e em pacientes com MG autoimune adquirida e avaliar a validade do método nas formas generalizada e ocular da doença. MÉTODOS: Foram estudados 20 sujeitos saudáveis, 20 pacientes com miastenia gravis forma generalizada (grupo MGG) e 13 com a forma ocular da doença (grupo MGO). A EMGFU-JAC foi realizada em todos os participantes, idealmente com 20 medidas de jitter em cada estudo. O jitter foi expresso como a média das diferenças consecutivas (MCD). Em todos os pacientes do estudo foram realizados o teste de estimulação repetitiva e dosagem sérica de anticorpo antirreceptor de acetilcolina (ac-AChR) no momento da análise do jitter. Nos pacientes soronegativos para ac-AChR, foi pesquisado o anticorpo antimúsculo específico tirosina-quinase (ac-MuSK). Foram definidos o limite superior da normalidade (LSN) para a média do MCD de cada estudo e para valores individuais de MCD. Os critérios de anormalidade foram: (1) média do MCD acima do LSN; ou (2) mais de 10% dos valores individuais de MCD acima do LSN. A definição do LSN para valores individuais de MCD baseou-se no conceito de que dois entre 20 valores de MCD acima do LSN são aceitáveis em um músculo saudável, para a técnica de contração voluntária. Portanto, estimou-se o LSN para o 18o valor mais alto de MCD (18o par). Para análise da acurácia do método, foram construídas duas curvas ROC (Receiver Operating Characteristic) para as variáveis média do MCD e 18o par, no grupo de pacientes (MGG e MGO) versus controle. RESULTADOS: No grupo controle a média das médias do MCD foi (19,0 ± 2,4)us e a média do 18o valor mais alto de cada estudo foi (24,5 ± 3,6)us. Esses valores obtidos apresentaram distribuição Gaussiana e o LSN foi definido como a média desses valores + 2 DP. O LSN para a média do MCD foi 24us, e 32?s para valores individuais de MCD. No grupo MGG, a análise do jitter foi anormal em todos os 20 pacientes por ambos os critérios de anormalidade, exceto em um paciente que apresentou anormalidade por apenas um dos critérios. No grupo MGO, apenas um dos 13 pacientes não preencheu os critérios de anormalidade. No grupo de pacientes, a positividade da EMGFU-JAC foi maior do que o teste de estimulação repetitiva e dosagens de anticorpos. Nas curvas ROC para as variáveis médias do MCD e 18o par, o valor de melhor sensibilidade (93,9%), sem resultados falsos positivos, foi 24,7us e 33,1us, respectivamente. CONCLUSÕES: A EMGFU-JAC apresenta alta sensibilidade e especificidade na identificação de distúrbio da transmissão neuromuscular em pacientes com MG. A utilização da agulha concêntrica é válida para a análise do jitter, como alternativa à agulha de fibra única / INTRODUCTION: Single fiber electromyography (SFEMG) technique, through jitter analysis, is the most sensitive neurophysiological method for confirmation of neuromuscular junction disorder in myasthenia gravis (MG). Records are traditionally obtained with single fiber needle, which is reusable and has a high-cost. Due to the current need of using disposable material, concentric needle has been used to replace single fiber needle. The technique is similar, but the action potential for jitter analysis is obtained with concentric needle electrode (SFEMG - concentric needle jitter, SFEMG-CNJ). However, studies are scarce and methodologies used are heterogeneous with the use of this needle. OBJECTIVES: This study aims to measure jitter values obtained with concentric needle in the Orbicularis Occuli muscle in healthy subjects and in patients with autoimmune acquired MG and to assess the validity of the method in generalized and ocular forms of the disease. METHODS: 20 healthy subjects, 20 patients with generalized myasthenia gravis (GMG group) and 13 with the ocular form of the disease (OMG group) were studied. SFEMG-CNJ was performed on all participants, ideally with 20 jitter values in each study. Jitter was expressed as the mean consecutive difference (MCD). Repetitive nerve stimulation and serum acetylcholine receptor antibody (AChR-ab) were performed in all patients in the study, by the time of jitter analysis. Tyrosine kinase specific antibody muscle antibodies (MuSK-ab) were performed in AChR-ab negative patients. The upper limit of normality (ULN) for the mean MCD and for individual jitter values were defined. The abnormality criteria were: (1) mean MCD above ULN; or (2) more than 10% of individual jitter values above ULN. The definition of ULN for individual jitter values was based on the concept that two out of 20 jitter values above ULN are acceptable in a healthy muscle for voluntary contraction technique. Therefore, the ULN for the 18th highest jitter value (18 pair) was estimated. To analyze the method\'s accuracy, two ROC curves (Receiver Operating Characteristic) for the mean MCD and 18th pair in the group of patients (MGG and MGO) versus control were constructed. RESULTS: In the control group the mean of MCD means was (19.0 ± 2.4)us and the mean of the 18 highest value of each study was (24.5 ± 3.6)us. These values showed Gaussian distribution and the ULN was set as the mean of these values + 2 SD. The ULN for the mean MCD was 24us, and 32us for individual values of MCD. In GMG group, jitter analyses were abnormal in all 20 patients based on both abnormality criteria, except in one patient, who had abnormalities in only one of the criteria. In OMG group, only one patient from 13 met neither of the abnormality criteria. In patients, the positivity of SFEMG-CNJ was higher than repetitive nerve stimulation test and antibody detection. The ROC curve threshold showing the best sensitivity (93.9%) with no false positive results was 24.7Us for the mean MCD and 33.1us for individual pairs, respectively. CONCLUSIONS: SFEMG-CNJ has high sensitivity and specificity in identifying neuromuscular transmission disorder in patients with MG. The use of concentric needle is valid for jitter analysis as an alternative to single fiber needle

O estudo do controle neurofisiológico do movimento tem sido realizado sob várias perspectivas. Experimentos com seres humanos são realizados durante a execução de uma dada tarefa motora e, frequentemente, mediante a aplicação de estímulos externos (elétrico, magnético ou mecânico) ao sistema neuromuscular. Estes experimentos fornecem uma grande quantidade de dados referentes ao funcionamento das redes neuronais e dos atuadores biomecânicos envolvidos nos procedimentos. Entretanto, alguns achados experimentais permanecem incompreensíveis, requerendo a utilização de outros recursos para elucidar quais mecanismos estão por trás dos resultados. Neste sentido, a modelagem matemática e a simulação computacional servem como parte importante destas ferramentas que são imprescindíveis para uma melhor compreensão dos mecanismos neurofisiológicos e biomecânicos por trás do controle do movimento. A presente tese de doutorado teve como objetivo prover um modelo neuromusculoesquelético biologicamente plausível capaz de investigar diferentes mecanismos responsáveis pelo controle do torque gerado na articulação do tornozelo. Este modelo teve como base um modelo neuromuscular previamente proposto, porém, que não incorporava uma série de elementos fundamentais para um estudo mais amplo do sistema motor. O novo modelo proposto contempla modelos de motoneurônios com dendritos ativos, proprioceptores musculares responsáveis pelas vias reflexas de curta e média latência, modelos que representam as características viscoelásticas dos músculos e um modelo biomecânico do ser humano durante a postura ereta quieta. O modelo foi aplicado a diferentes problemas relacionados ao funcionamento do sistema neuromusculoesquelético, que são tipicamente explorados por experimentos com seres humanos, e forneceu bases teóricas importantes para estes achados. / The neurophysiological control of movement has been studied from several standpoints. Human experiments are performed during the execution of a given motor task and, frequently, by applying an external stimulation (electrical, magnetic, or mechanical) to the neuromuscular system. These experiments provide a large amount of data concerning the functioning of the neuronal networks and biomechanical actuators involved in the procedures. Nonetheless, some experimental findings remain puzzling, so that other available resources should be used to clarify what mechanisms are behind these results. In this vein, the mathematical modeling and computer simulations are invaluable tools that may be used to better understand the neurophysiological and biomechanical mechanisms underlying the motor control. The present PhD thesis aimed at providing a biologically plausible neuromusculoskeletal model that was used to study different mechanisms involved in the control of the ankle joint torque. This model was based on a previous neuromuscular model, which did not employ several elements that are fundamental to a comprehensive evaluation of the motor system. The novel proposed model encompasses motor neuron models with active dendrites, muscle proprioceptors responsible for the short- and medium-latency reflex pathways, muscle models with the main viscoelastic features, and a biomechanical model of the human body during upright stance. It was applied to a series of problems frequently related to the functioning of the neuromusculoskeletal system and its main outcomes provided important theoretical bases for a set of experimental findings.

The greatest genetic factor in how well we age cognitively is Apolipoprotein E (APOE), a single nucleotide polymorphism with three allelic variants: epsilon-2, epsilon-3 and epsilon-4 (hereafter ε2, ε3, ε4). The ε4 allele is associated with an increased risk of cognitive disadvantage in later life, however, the effects of this variant are not isolated to old-age, with some studies reporting cognitive advantages in youth. This thesis investigates the influence of APOE ε4 on cognition from mid-adulthood, a point in the lifespan when the detrimental effects of this allele may be emerging. This thesis begins with a systematic review and meta-analysis of the literature to-date, and suggests attention may be sensitive to ε4 differences in mid-adulthood, however, effects of the allele are not consistently shown, perhaps due to methodological limitations including the use of insensitive neuropsychological batteries (Chapter 1). Next, behavioural paradigms providing a sensitive index of both selective (Chapter 2) and executive attention (Chapter 3), suggest many attentional processes are intact in mid-age (45-55 years) ε4 carriers. Subtle deficits, however, are apparent on prospective memory (PM) and Stroop-switch paradigms, indicating a goal maintenance disadvantage. In addition, a proxy of cognitive reserve was found to moderate the effects of ε4 on executive attention in mid-adulthood (Chapter 4). Follow-up research used paradigms that target the distinct processes supporting focal and non-focal PM to interrogate the profile of change observed in mid-age ε4 carriers, identifying a profile of disadvantage consistent with that observed in pathological ageing (Chapter 5). PM, however, was not found to differentiate ε4 carriers in older individuals at heightened risk of converting to dementia (Chapter 6). Collectively, this research provides evidence for a profile of accelerated ageing in ε4 carriers, with subtle disadvantages apparent in executive attention by the end of the 5th decade.