L’autophagie est un processus catabolique lié aux lysosomes. L’autophagie joue un rôle dans la biologie des lymphocytes et dans la réponse immunitaire en générale. Nous avons montré une dérégulation de l’autophagie dans les lymphocytes provenant de souris développant un lupus et de patients atteints d’un lupus érythémateux disséminé. Nous avons ensuite cherché à définir le rôle potentiel de l’autophagie des lymphocytes dans l’activation et le maintien des réponses humorales normales et pathologiques. Ainsi, nous avons généré des souris déficientes en autophagie spécifiquement dans les lymphocytes B. Ces modèles de souris nous ont permis de montrer que l’autophagie ne jouait pas de rôle majeur dans la mise en place de la réponse immunitaire humorale à court terme. Cependant, l’étude du même modèle murin sur fond génétique prédisposant à une auto-immunité systémique a démontré un rôle de l’autophagie dans la production d’auto-anticorps anti-nucléaires et dans le maintien d’un fort nombre de plasmocytes. L’autophagie est donc importante pour l’initiation de l’activation des lymphocytes B et leur survie en contexte d’auto-immunité à long terme. Ce modèle nous a également permis de montrer que l’absence d’autophagie lors de la stimulation du BCR compromet sa polarisation, conjointe à celle des molécules complexe majeur d’histocompatibilité de classe II et des lysosomes. Ce phénomène est important dans la mise en place de la synapse immunologique, structure qui permet la dégradation et l’internalisation d’antigène particulaires. Nous avons pu mettre en évidence que l’inhibition de l’autophagie impacte effectivement la présentation d’antigènes particulaires internalisés via le BCR aux lymphocytes T. Ainsi, la modulation de l’autophagie dans les lymphocytes pourrait permettre à plusieurs niveaux de limiter l’activation et la survie des lymphocytes autoréactifs dans le contexte de maladies auto-immunes. / Macroautophagy, called autophagy, is a catabolic lysosomal process. Macroautophagy was recently shown to regulate the immune response especially by regulating lymphocyte biology. We demonstrated that autophagy is deregulated in T cells from lupus mouse models and patients suffering from systemic lupus erythematosus. We suggest that autophagy could regulate the survival of autoreactive lymphocytes during lupus. We then wanted to better understand the role of autophagy in normal and pathologic humoral responses. We have generated mouse models conditionally deficient for ATG5 in B cells. In accordance with previous studies, we show that autophagy is dispensable for B cell survival and activation under short-term B cell receptor (BCR) activation. We then investigated long-term immunity on a spontaneous model of autoimmunity. In autoimmune-prone mice deficient for autophagy in B cells, we demonstrate that autophagy is important to maintain high levels of anti-nuclear auto-antibodies, and high number of long-lived plasma cells in the bone marrow. With these same mouse models, we show that autophagy contributes to the polarization of internalized BCR after stimulation, together with the recruitment of lysosomes and MHCMII molecules-containing compartments. The polarization of B cells is particularly important for the acquisition of particulate antigens for B cells. We postulate that ATG5 and possibly the autophagic machinery could facilitate the formation of the immune synapse. We indeed demonstrate that presentation of immobilized antigens to T cells is compromised in the absence of ATG5 in B cells. Thus, modulating autophagy in lymphocytes, could limit at several levels the activation and/or survival of autoreactive lymphocytes during autoimmunity.

Submitted by Luciana Sebin (lusebin@ufscar.br) on 2016-09-20T17:56:33Z No. of bitstreams: 1 TeseWLO.pdf: 4950921 bytes, checksum: adf4e51b260eb25ef34f14a351022af5 (MD5) / Approved for entry into archive by Marina Freitas (marinapf@ufscar.br) on 2016-09-21T12:48:10Z (GMT) No. of bitstreams: 1 TeseWLO.pdf: 4950921 bytes, checksum: adf4e51b260eb25ef34f14a351022af5 (MD5) / Approved for entry into archive by Marina Freitas (marinapf@ufscar.br) on 2016-09-21T12:48:16Z (GMT) No. of bitstreams: 1 TeseWLO.pdf: 4950921 bytes, checksum: adf4e51b260eb25ef34f14a351022af5 (MD5) / Made available in DSpace on 2016-09-21T12:48:30Z (GMT). No. of bitstreams: 1 TeseWLO.pdf: 4950921 bytes, checksum: adf4e51b260eb25ef34f14a351022af5 (MD5) Previous issue date: 2016-01-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / In this thesis, a wide general class of models for mixed responses is proposed in which joint distributions are constructed by the conditional approach (probability density functions, (pdf), as the product of a marginal pdf and a conditional pdf). It is assumed that the distribution of the discrete response and the conditional distribution of the continuous response given the discrete variable belong to one- or two-parameter exponential family of distributions. Furthermore, the marginal means are related to the covariates by link functions using linear and/or nonlinear and/or non-parametric predictors and a dependency structure between the responses is introduced into the model via the conditional mean. Estimation methods, diagnostic analysis and in uence techniques are presented as well as a simulation study considering the Bernoulli-exponential and Poisson-normal semiparametric models, two particular cases of the proposed class. Finally, one of the proposed models is used in a real data set involving the total cost of care for each patient during hospitalization, the use or not of the intensive treatment units and the age of the patient. / Nesta tese é proposta uma classe ampla e geral de modelos bivariados para respostas mistas em que as distribuições conjuntas são construídas pelo método da fatoração (função densidade de probabilidade, (fdp), como o produto de uma fdp marginal e uma fdp condicional). É assumido que a distribuição da variável resposta discreta e a distribuição condicional da variável resposta contínua dada a variável discreta pertencem à família exponencial de distribuições uniparamétrica ou biparamétrica. Além disso, as médias marginais são relacionadas a covariáveis através de funções de ligação usando preditores linear e/ou não linear e/ou não paramétrico e uma estrutura de dependência entre as respostas é inserida no modelo via a média condicional. Métodos de estimação, análises de diagnóstico e técnicas de in uência são apresentadas assim como um estudo de simulação considerando os modelos Bernoulli-exponencial e Poisson-normal semiparamétrico, dois casos particulares da classe proposta. Finalmente, um dos modelos propostos é usado em um conjunto de dados reais envolvendo gastos totais com cuidados para cada paciente durante a hospitalização, o uso ou não da unidade de tratamento intensivo e a idade do paciente.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The research of neural mechanisms related to training of emotions has increased in recent years. Fear is a behavior originated in response to the dangers encountering by animals, originating in defensive responses displayed when exposed to threatening stimuli. Fear memory helps animals and humans recognize putative sources of danger and adopt the appropriate behavioral response. The primary neural circuits for fear acquisition and extinction involve connections between prefrontal cortex, ventral hippocampus and amygdala, and these áreas are modulated by brainstem networks. The nucleus (n.) incertus in the dorsal pontine tegmentum provides a strong GABAergic projection to these forebrain centers and is strongly activated by neurogenic stressors. In this study in male, adult rats, we injected miniruby anterograde tracer into n. incertus and delineated its projections to the amygdala; and examined the effect of electrolytic lesions of n. incertus on different stages of the fear conditioning-extinction process. N. incertus-derived nerve fibers were observed in anterior medial amygdala, endopiriform nucleus, intra-amygdala bed nucleus of stria terminalis, amygdalohippocampal transition area, and the ventromedial nucleus of the lateral amygdala, with a broad fiber band present between the basolateral amygdala and the olfactory nuclei of amygdala. In a conventional contextual fear conditioning paradigm, we compared freezing behavior in control (naïve) rats (n = 13), with that in rats after sham- or electrolytic lesions of n. incertus (n = 9/group). There were no differences between the three groups in the habituation, acquisition, or context conditioning phases; but n. incertus-lesioned rats displayed a markedly slower (delayed) extinction of conditioned freezing responses than sham/control rats; suggesting n. incertus-related circuits normally promote extinction through inhibitory projections to amygdala and prefrontal cortex. The results helps in understanding the neurobiological mechanisms involved and in development of the future biotechnological techniques to minimize the effects of disorders associated with fear in humans, such as panic, anxiety and posttraumatic stress disorder. / A pesquisa dos mecanismos neurais relacionados à formação das emoções tem crescido nos últimos anos. O medo é um comportamento originado em resposta aos perigos enfrentados pelos animais, tendo sua origem nas reações defensivas exibidas quando da exposição a estímulos ameaçadores. A memória do medo ajuda os animais e os seres humanos a reconhecem as fontes putativas de perigo e adotar a resposta comportamental apropriada. Os circuitos neurais primárias envolvidos nos mecanismo de aquisição de medo e extinção envolvem conexões entre o córtex pré-frontal, hipocampo ventral e a amígdala, e estas áreas são moduladas por redes do tronco cerebral. O núcleo incertus (NI) no tegmento dorsal pontino fornece uma forte projeção GABAérgica a estes centros prosencéfalicos e é fortemente ativado por estressores neurogênicos. Neste estudo em ratos adultos machos foi injetado o traçador anterógrado miniruby no NI, delineado as suas projeções para a amígdala e examinado o efeito de lesões eletrolíticas no NI sobre diferentes fases do processo de condicionamento do medo-extinção. Fibras derivadas do NI foram observadas na amígdala medial anterior, núcleo endopiriforme, parte intra-amígdala do núcleo do leito da estria terminalis, área de transição amígdala-hipocampal, e o núcleo ventromedial da amígdala lateral, com uma ampla faixa de fibra presentes entre a amígdala basolateral e os núcleos olfativos da amígdala. Em um paradigma de condicionamento contextual de medo convencional, comparou-se o comportamento de congelamento em ratos controle (não operados) (n = 13), com ratos operados sem lesão do NI e ratos com lesão do núcleo incertus (n = 9). Não houve diferenças entre os três grupos nas fases de habituação, aquisição ou condicionamento ao contexto, mas ratos com lesão no NI exibiram uma extinção nitidamente mais lenta (com atraso) de respostas condicionadas de congelamento em comparação com ratos operados sem lesão do NI e controles, sugerindo que circuitos NI relacionados normalmente promovem a extinção através de projeções inibitórias para a amígdala e o córtex pré-frontal. Os resultados encontrados auxiliam na compreensão dos mecanismos neurobiológicos envolvidos e no desenvolvimento futuro de técnicas terapêuticas biotecnológicas visando minimizar os efeitos dos distúrbios associados ao medo em humanos, a exemplo do pânico, ansiedade patológica e estresse pós-traumático.

The production of cereals, especially maize, which is a staple food in sub-Saharan Africa, is challenged by pests and diseases. In addition, climate change will exacerbate the magnitude of these challenges and agriculture in general. Lepidopteran stemborers are major pests of cereal crops in sub-Saharan Africa. Two indigenous noctuids, Busseola fusca and Sesamia calamistis and one exotic crambid, Chilo partellus, occur as single and mixed-species communities infesting cereal crops in East and southern Africa. The composition of these communities however varies with altitude and over seasons. Interactions between the species especially when in combination, can affect the level of damage to cereal crops. The objective of this study was to describe the intraspecific and interspecific interactions between the three stemborer species when they make use of the same resources. This study involved field surveys in maize fields in major agro-ecological zones and also different experiments under greenhouse and laboratory conditions to describe the interactions between the species as well as the effect of temperature, species density and duration of the period of competition on outcomes of these interactions. Results showed that stemborer communities are composed of single and mixed species of B. fusca, S. calamistis and C. partellus, which varied with the different agro-ecological zones and also along altitudinal gradients. Temperature was the most important abiotic factor that influenced the composition of stemborer communities. However, infestation patterns varied with season. This study also showed that female moths of the three species did not avoid oviposition on plants that were previously infested by stemborers. Busseola fusca showed a high preference for heterospecific-infested plants, while C. partellus preferred conspecific-infested plants. Sesamia calamistis did not show a significant preference for one plant over the other. This study further showed that both intra- and interspecific competition characterise communities of these stemborer species which ultilise the same resources. Interspecific competition was stronger between the noctuids and the crambid than between the two noctuid species. Temperature had a significant influence on the competitive outcomes between the three species. Finally the study showed that densitydependent effects and the duration of the interactions are also important factors which influence the level of competitive outcomes between the species. Climate change is likely to influence the interactions and composition of stemborer communities. / PhD (Environmental Sciences), North-West University, Potchefstroom Campus, 2015

The production of cereals, especially maize, which is a staple food in sub-Saharan Africa, is challenged by pests and diseases. In addition, climate change will exacerbate the magnitude of these challenges and agriculture in general. Lepidopteran stemborers are major pests of cereal crops in sub-Saharan Africa. Two indigenous noctuids, Busseola fusca and Sesamia calamistis and one exotic crambid, Chilo partellus, occur as single and mixed-species communities infesting cereal crops in East and southern Africa. The composition of these communities however varies with altitude and over seasons. Interactions between the species especially when in combination, can affect the level of damage to cereal crops. The objective of this study was to describe the intraspecific and interspecific interactions between the three stemborer species when they make use of the same resources. This study involved field surveys in maize fields in major agro-ecological zones and also different experiments under greenhouse and laboratory conditions to describe the interactions between the species as well as the effect of temperature, species density and duration of the period of competition on outcomes of these interactions. Results showed that stemborer communities are composed of single and mixed species of B. fusca, S. calamistis and C. partellus, which varied with the different agro-ecological zones and also along altitudinal gradients. Temperature was the most important abiotic factor that influenced the composition of stemborer communities. However, infestation patterns varied with season. This study also showed that female moths of the three species did not avoid oviposition on plants that were previously infested by stemborers. Busseola fusca showed a high preference for heterospecific-infested plants, while C. partellus preferred conspecific-infested plants. Sesamia calamistis did not show a significant preference for one plant over the other. This study further showed that both intra- and interspecific competition characterise communities of these stemborer species which ultilise the same resources. Interspecific competition was stronger between the noctuids and the crambid than between the two noctuid species. Temperature had a significant influence on the competitive outcomes between the three species. Finally the study showed that densitydependent effects and the duration of the interactions are also important factors which influence the level of competitive outcomes between the species. Climate change is likely to influence the interactions and composition of stemborer communities. / PhD (Environmental Sciences), North-West University, Potchefstroom Campus, 2015

Cochlear implants (CIs) and auditory brainstem implants (ABIs) are prostheses for hearing used in patients with profound hearing impairment. A CI requires an operational cochlear nerve to function in contrast to an ABI. ABIs were initially designed for adult patients with neurofibromatosis type 2 (NF2), suffering from bilateral vestibular schwannomas. Now ABIs are also used for patients, both adults and children, with congenital cochlear malformations, cochlear nerve hypoplasia/aplasia, and cochlear ossification. The aims of this thesis are to evaluate hearing outcome in patients implanted with a CI after long-term deafness. An extended period of deafness has earlier been considered as a contraindication for CI surgery. Further, we analyzed if electrically evoked auditory brainstem responses (eABRs) can predict CI outcome and pinpoint the optimal selection of treatment such as CI or ABI. We also disclose our experiences from ABI surgery in Uppsala, such as implant use, hearing outcome, complications, and satisfaction among the patients. Finally, we evaluated the results and benefits of ABIs in non-NF2 pediatric patients. Results show that patients with an extended deafness period and durations over 20 years can achieve speech understanding and benefit from CIs. Patients with long-term deafness and limited years of hearing before deafness did not perform as well as those with shorter deafness duration and longer hearing experience did. eABR seems to have a definite role in the diagnostic armamentarium, to better consider alternative surgical strategies such as ABI. No eABR waveform predicted a poor CI outcome. There was no correlation between speech perception and eABR waveform latencies or eABR waveform quality. A majority of the ABI patients used their ABIs and benefited from them for at least some period. ABI assisted voice control in a majority of the full-time users and they reported improved understanding of speech with the implant switched on. No severe complications from ABI surgery or ABI stimulation were noted. The patients were generally satisfied, even if their hearing remained very limited. All pediatric patients but one used the implant continuously and benefited from it.

Zusammenfassung Die europäische Buche (Fagus sylvatica, L.) ist eine dominante Waldbaumart von hohem ökonomischen und ökologischem Wert in Zentraleuropa. Die natürliche Verbreitung der Spezies in Zentraleuropa ist abhängig von der Wasserverfügbarkeit. In der Zukunft wird vermehrt von extremen Wetterbedingungen wie Hitzewellen und extreme Trockenheit ausgegangen. In ausgedehnten Bereichen des Waldes wird die Wasserversorgung wahrscheinlich ein limitierender Faktor. Daher ist es notwendig, die potentielle Fähigkeit der Buchen, sich bei Wasserlimitierung zu akklimatisieren oder anzupassen, zu bewerten. Die Reaktionen der Buche auf Wasserknappheit könnten in anatomischen Eigenschaften des Holzes abgespeichert sein und durch die Analyse dieser Eigenschaften bewertet werden. Darüber hinaus könnte die Wasserlimitierung die Kohlenstoff- und Stickstoffgehalte im Buchenholz negativ beeinflussen. Die Bodenfeuchte ist ein wichtiger Faktor, der den 13C-Gehalt in den Baumringen beeinflußt. Da Wasserknappheit den Verschluß der Stomata induzieren kann, wird das δ13C des eingebauten Kohlenstoffes erhöht. Daher wurden C- und N-Gehalte und δ13C Signaturen in Buchenholzproben analysiert, um die Auswirkungen von Wasserknappheit auf Buchenholzeigenschaften zu untersuchen. Eine Schlüsselrolle für die Akklimatisierung an Trockenheit spielt Abscisinsäure (ABA), wodurch Abwehrreaktionen hervorgerufen werden, die zum Verschluß der Stomata führen und somit den Wasserverbrauch der Pflanzen regulieren. Ein weiteres Merkmal von Trockenstress ist eine erhöhte Produktion von reaktiven Sauerstoffspezies. Daher ist die Aktivierung von Schutzenzymen, insbesondere der antioxidativen Abwehr, wichtig bei der Bekämpfung des oxidativen Abbaus anfälliger Strukturen wie etwa der Zellmembranen. Um die Plastizität und Anpassung der Buche in Reaktion auf Trockenheit zu untersuchen, wurde die Expression von ABA- und stressverwandten Genen für die Analyse ausgewählt. Darüber hinaus wurden die Blattflächen und die Membranintegrität als Indikatoren für Reaktionen der Buche auf Trockenstress bestimmt. Pflanzen haben unterschiedliche Strategien, um Trockenstress zu bewältigen: Vermeidung oder Toleranz. Der Grundmechanismus beider Strategien beinhaltet die isohydrische oder anisohydrische Regulation der Stomata. Isohydrische Pflanzen schließen ihre Stomata noch bevor sich der Wasserstatus in der Pflanze verändert, wohingegen anisohydrische Spezies eine langsame stomatale Reaktion als Antwort auf ein geringeres Wasserpotential zeigen. Der Wassergehalt des Bodens, in den Blättern vor Sonnenaufgang (Predawn Water Potential), der relative Wassergehalt, Chlorophyllfluoreszenz und stomatäre Leitfähigkeit wurden als gute Merkmale charakterisiert, um diese Strategien zu testen. Des Weiteren wurde die Expression von OST1 (open stomata 1), einer Proteinkinase, die zur Schließzellenreaktion des ABA-Signalnetzwerkes führt, untersucht. In dieser Studie wurden die Reaktionen von Sämlingen und jungen, sowie ausgewachsenen europäischen Buchen auf Trockenheit untersucht. Die folgenden Hypothesen wurden getestet: • Buchenpopulationen aus trockeneren Lebensräumen weisen Veränderungen in der Anatomie des Xylems auf, um mit geringem Niederschlag umzugehen. • Trockenes Klima wirkt sich negativ auf den Kohlenstoff und den Stickstoffgehalt in Buchenholz aus. • Buchennachkommen von trockenen Standorten zeigen eine konstitutiv erhöhte Expression von ABA-und stressinduzierten Genen und reagieren somit weniger auf Trockenheit als Nachkommen von feuchteren Standorten. • Buchen, die aus niederschlagsarmen klimatischen Bedingungen stammen, zeigen eine stärkere Trockenheitsvermeidung. Buchen aus mesischen Habitaten bilden eine stärkere Toleranz gegenüber Trockenheit aus, als solche aus trockenen Habitaten, wenn sie einer abnehmenden Wasserverfügbarkeit in der Erde ausgesetzt sind. Um diese Hypothesen zu testen, wurden drei Experimente durchgeführt. Einerseits mit adulten Buchen entlang eines Niederschlagsgradienten, andererseits mit Buchensetzlingen, die experimentell veränderten Bodenwassergehalten ausgesetzt waren. Ein Freilandexperiment wurde in drei verschiedenen Gebieten durchgeführt, die ähnliche Bodeneigenschaften aufwiesen, sich aber hinsichtlich der jährlichen Niederschlagsrate unterschieden. Holzzuwachs, die anatomischen Eigenschaften des Xylems, sowie C- und N- Gehalt und die δ13C Signaturen wurden untersucht. Es wurde eine starke Reduzierung des jährlichen Zuwachses bei Buchen von feuchten hin zu trockenen Standorten gefunden. So könnte die Verfügbarkeit von Wasser in den Untersuchungsgebieten einer der begrenzenden Faktoren des Holzzuwachses bei Buchen sein. Buchen auf trockenen Standorten zeigten Veränderungen anatomischer Merkmale, die ihnen ermöglichten, besser mit geringen Niederschlägen umzugehen. Um schmale Gefäßlumen auszugleichen, zeigten Buchen an trockenen Standorten mehr Gefäße. Diese anatomischen Veränderungen ermöglichen Buchen wahrscheinlich die Balance zwischen Wasseraufnahmeeffizienz und der Vermeidung von Embolien im Buchenstamm zu halten. Darüber hinaus trägt dieser Mechanismus wahrscheinlich dazu bei, das Wasserpotential und den C- und N-Gehalt im Holz der Buchen unter trockenen Bedingungen zu erhalten. Dieses Ergebnis deutet darauf hin, dass Buchen von trockenen Standorten eine Vermeidungsstrategie gegen Trockenheit haben, um mit geringer Wasserverfügbarkeit in der Natur umgehen zu können. Anatomische Merkmale variierten während der Vegetationsperiode signifikant. Zwischen den Standorten wiesen die anatomischen Parameter im Frühholz keine bemerkenswerten Veränderungen auf. Im Spät- und Übergangsholz war die Fläche der Gefäßlumen stark vermindert und die Anzahl der Gefäße signifikant erhöht. Im Spätholz der Buchen auf trockenen Standorten wurden dickere Wände und schmalere Faserlumina gefunden. Darüber hinaus zeigten verringerte δ13C Werte bei den Buchen auf den trockensten Standorten eine höhere Wassernutzungseffizienz am Ende der Wachstumsperiode. Der Vergleich der Bäume von feuchten und von trockenen Standorten wies darauf hin, dass die Verfügbarkeit von Wasser anatomische Veränderungen beeinflußte. Jedoch können auch andere Faktoren als die genetischen zu einer besseren Anpassung der Buchen auf den trockenen Standorten an geringe Niederschläge beitragen. Um die Expression von Genen im Zusammenhang mit ABA und Stress als Reaktion auf Trockenstress zu untersuchen, wurde ein Gartenexperiment durchgeführt. Für dieses Experiment wurden die Nachkommenschaften von fünf Buchenbeständen entlang eines Niederschlagsgradienten verwendet. Die Reaktionen von gut bewässerten und trockengestressten Keimlingen gegenüber Trockenstress wurden während des Sommers zu einem frühen, mittleren und späten Zeitpunkt gemessen. Die Expressionsniveaus von ABA- und stressbezogenen Genen wurde ermittelt. Um die Genexpression mit der Leistungsfähigkeit der Pflanzen vergleichen zu können, wurden herkunfts- und dürrebedingte Auswirkungen auf die Blattfläche und Membranintegrität in Abwesenheit und Anwesenheit von akutem oxidativen Stress untersucht. Trockenstress führte zu einer verringerten Blattfläche verglichen mit gut gewässerten Setzlingen. Die Nachkommen von feuchteren Standorten zeigten allgemein größere Blattflächen als die von trockenen Standorten. Der relative Verlust von Elektrolyten wurde durch Trockenstress verändert und erhöhte sich gegen Ende der Vegetationsperiode. Die Expressionsniveaus von ABA- und stressverwandten Genen wurden stark von Trockenstress beeinflußt. Eine Ausnahme bildet die Glutamin Amidotransferase (GAT). Zusätzlich waren die Expressionsniveaus der Gene Nine cis-Epoxy Dioxygenase (NCED), Proteinphosphatase 2C (PP2C), Early Responsive to Dehydration (ERD), Ascorbat-Peroxidase (APX), Superoxid Dismutase (Cu / Zn-SOD), Aldehyde Dehydrogenase (ALDH), Glutamin Amido-Transferase (GAT) höher in den Nachkommen von feuchten Standorten verglichen mit trockeneren Standorten. Saisonale Analysen der transkriptomalen Regulation von Genen für die Signalisierung und Abwehr von Trockenheit zeigten intraspezifische Unterschiede in der konstitutiven Expression und Reaktionsfähigkeit bei Trockenheit. Die herkunftsbedingten Unterschiede waren größer als die Stressreaktionen, was darauf hindeutet, dass die Selektion für eine Anpassung an Trockeheit bereits in lokalen Buchenpopulationen stattfindet. Um zu untersuchen, ob es intraspezifische Unterschiede bei den Resistenzmechanismen gegen Trockenheit gibt, wurden drei Buchenherkünfte, aus einem niedrigen, einem mittleren und einem hohen Niederschlagsklima (als LP, IP und HP bezeichnet), zunehmender Trockenheit ausgesetzt. Der Wassergehalt in Boden und Pflanzen, die maximale Quantenausbeute des Photosystems II und die stomatäre Leitfähigkeit der Kontrollen und der trockenheitsbehandelten Setzlinge wurden regelmäßig gemessen. Außerdem wurden die Transkriptionsniveaus von OST1 bestimmt. Die Daten weisen darauf hin, dass die innerartlichen Reaktionen auf Trockenheit bei Buchen auch zwischen isohydrischem und anisohydrischem Verhalten der Stomata variieren können. Die Buchenherkunft LP zeigte einen isohydrischen Phänotyp, da die Pflanzen einen schnelleren Verschluß der Stomata, einen höheren relativen Wassergehalt, sowie einen höheren Wassergehalt in den Blättern vor Sonnenaufgang (Predawn Water Potential) zeigten als Buchennachkommen aus mesischen Bedingungen. Dadurch wies die Population aus dem trockenen Habitat eine deutliche Vermeidungsstrategie bei Trockenheit auf. Im Gegensatz dazu zeigten die HP Nachkommen bei Wasserlimitierung einen langsamen Abfall der stomatären Leitfähigkeit, jedoch eine stärkere Abnahme des Predawn Water Potential. Es gab keinen Einfluss durch Trockenheit auf das Pflanzenwachstum oder die Biomasseallokation während der Trockenheitsbehandlung. Buchen zeigten intraspezifische Unterschiede bei den Resistenzstrategien gegen Trockenheit, gekennzeichnet durch anisohydrisches oder isohydrisches Verhalten. Das legt nahe, dass der anisohydrische Funktionstyp bei Buchen besser geeignet ist, um mit den vorhergesagten Klimaextremen umzugehen, als die isohydrische Typ, da er eine Trockentoleranz-Strategie aufweist. Die Ergebnisse der vorliegenden Studie zeigen, dass Klimate mit geringen Niederschlägen und Trockenheit die Anatomie, Physiologie und molekulare Reaktionen von Buchen beeinflussen. Buchen zeigten recht hohe intraspezifische Unterschiede bei den Strategien zur Trockenheitsresistenz, mit Strategien zur Trockenheitsvermeidung und Trockentoleranz.

The detection of predators by prey species is crucial in order to escape the threat posed by a predator. In mammals, the olfactory sensory system is commonly used to detect odors emitted by predators, and to determine how threatening the situation actually is. However, knowledge about this ability is still sparse and in some cases conflicting. The aim of the present study was therefore to assess whether CD-1 mice (Mus musculus) show behaviors such as avoidance, anxiety and/or decreased activity when exposed to any of the three odorants: cat bladder urine, horse voided urine or a fruity odor (N-pentyl acetate), with a blank solvent as an alternative in a two-compartment test arena. I found no significant differences between avoidance (the time that the mice spent in the different compartments), anxiety (the numbers of fecal pellets dropped by the mice), or the overall activity (the number of switches between the two compartments), when the mice were exposed to the three different odors. The fact that the cat urine derived from the bladder of the cat may explain the lack of avoidance responses, since bladder urine might not contain the same chemical components as voided urine. Bladder urine might therefore also lack the chemical components that signal “predator” to the mice. In conclusion, mice do not respond differently to the odor of cat bladder urine than to horse voided urine or to the fruity odor of N-pentyl acetate.

Thesis (MSc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: As ectotherms, lizards are particularly vulnerable to changes in the thermal landscape and face extinction risk if they lack the capacity to rapidly adapt or behaviourally mitigate increasingly altered thermal environments. Theoretical models that predict lizards‟ response to climate change often fail to take into account the thermal characteristics of the microenvironment, the ability of lizards to behaviourally buffer climate variation in the habitat and the plastic nature of both behaviour and physiology over ecologically relevant time-scales. Here, I address this major knowledge gap using two separate research chapters in an experimental physiology approach. In Chapter 1, I investigated the temperature-dependence and plasticity of resting metabolic rate, water-loss rate and preferred body temperature of Cordylus oelofseni at several temporal scales (within and between seasons) and incorporated field observations to acquire a better understanding of this species‟ adaptive potential to buffer thermal changes in the habitat. Cordylus oelofseni showed plasticity of both behaviour and physiology in response to thermal acclimation, but relied on distinct strategies depending on the time-scale investigated. These results highlighted the complexity of underlying mechanisms used by these organisms to buffer temperature variation. In Chapter 2, I used an experimental approach to examine the energetic costs of thermoregulation in C. oelofseni and test the cost-benefit model of thermoregulation. This model‟s primary prediction states that lizards should thermoregulate carefully only when the associated costs are low. Using four enclosures that simulated different thermal qualities (temporal and spatial distributions of operative temperatures) in the habitat, I found limited support for the cost-benefit model. Lizards in the low-quality heterogeneous enclosures invested the same energetic effort and thermoregulated with similar overall accuracy as lizards in the high-quality heterogeneous enclosure. The costs incurred were not necessarily energetic, but reflected missed opportunities (e.g. less time to forage), something that, along with important interaction effects with body mass, deserves further attention when testing this model. Together, these results illustrate the importance of incorporating ecological reality at various time and spatial scales in order to make relevant predictions regarding the fate of lizards with projected climate change. / AFRIKAANSE OPSOMMING: As ektotermiese diere, is akkedisse veral sensitief vir veranderinge in die termiese landskap en staar uitsterwingsrisiko in die gesig as hulle nie die vermoë het om vinnig aan te pas of gedragsveranderinge te maak in omgewings wat toenemend verwarm nie. Teoretiese modelle wat akkedisse se reaksie op klimaatsverandering voorspel, neem dikwels nie die termiese eienskappe van die mikro-omgewing, die vermoë van akkedisse om met gedragsveranderinge klimaat variasie in die habitat te buffer en die plastieke aard van beide gedrag en fisiologie oor ekologies relevante tydskale in ag nie. Hier bespreek ek hierdie groot kennisgaping met behulp van twee afsonderlike navorsingshoofstukke in 'n eksperimentele fisiologie benadering. In Hoofstuk 1 het ek ondersoek ingestel na die temperatuur-afhanklikheid en plastisiteit van rustende metaboliese tempo, waterverlies tempo en voorkeur liggaamstemperatuur van Cordylus oelofseni by verskeie tydskale (binne en tussen seisoene) en inkorporeer veld waarnemings om 'n beter begrip te verkry van hierdie spesie se aanpasbare potensiaal om termiese veranderinge in die habitat te buffer. Cordylus oelofseni het plastisiteit van beide gedrag en fisiologie in reaksie op hitte-akklimatisering getoon, maar staatgemaak op verskillende strategieë, afhangende van die tyd-skaal wat ondersoek is. Hierdie resultate beklemtoon die kompleksiteit van die onderliggende meganismes wat gebruik word deur hierdie organisme om temperatuur verandering te buffer. In Hoofstuk 2 het ek 'n eksperimentele benadering gebruik om die energiekoste van termoregulering in C. oelofseni te ondersoek en die kostevoordeel model van termoregulering te toets. Hierdie model se primêre voorspelling verklaar dat akkedisse slegs versigtig moet termoreguleer wanneer die gepaardgaande koste laag is. Deur gebruik te maak van vier afskortings wat verskillende termiese eienskappe gesimuleer het (tyd en ruimtelike verspreiding van operatiewe temperature) in die habitat, het ek beperkte ondersteuning gevind vir die koste-voordeel model. Akkedisse in die lae-gehalte heterogene afskortings het dieselfde energieke moeite belê en getermoreguleer met soortgelyke algehele akkuraatheid as akkedisse in die hoë-gehalte heterogene kamp. Die kostes wat aangegaan is, is nie noodwendig energiek nie, maar weerspieël geleenthede wat gemis is (bv. minder tyd om kos te soek), iets wat, saam met belangrike interaksie effekte met liggaamsmassa, verdere aandag verdien wanneer hierdie model getoets word. Tesame illustreer hierdie resultate die belangrikheid van die integrasie van ekologiese werklikheid op verskillende tyd en ruimtelike skale, om relevante voorspellings oor die lot van akkedisse met geprojekteerde klimaatsverandering te kan maak.

Mands, of which requesting and rejecting responses are considered subclasses, are the first emerging communication functions that allow children to express their wants and needs. While typically developing children develop speech without specifically designed intervention, many children with autism and developmental disabilities are likely to rely on prelinguistic communication forms that are socially and developmentally inappropriate or unacceptable until symbolic forms of functional communication are taught. A review of the literature on teaching mands indicates that although there is an abundance of research addressing teaching communicative requesting behaviors, rarely have studies attempted to teach communicative rejecting. The purpose of this study was to create rejecting opportunities using the wrong-item format embedded into the missing item format, and to teach socially appropriate rejecting response using AAC for four children with autism and developmental disabilities. This study employed a multiple probe design across four participants to examine the effectiveness of the procedure. Results indicated that the wrong-item format embedded into the missing-item format was effective in teaching symbolic forms of rejecting responses using VOCAs and PECS. The results were generalized across two untrained activities and were maintained up to four weeks following the termination of generalization probes for three participants. The implications and limitations of this study, as well as potential topics for future research are also discussed. / text