The aim of this diploma thesis is to identify options of how to use the systém of coordinated rehabilitation in a home with a special regime. To obtain information I used qualitative research methodology. Semi-standardized interviews served as the research tool. These interviews were carried out with 10 employees of a home with a special regime run by the City Institute of Social Services in Strakonice. The thesis is divided into a theoretical and a practical part. The theoretical part of the diploma thesis focuses on coordinated rehabilitation as a whole, further, it discusses the individual parts of coordinated rehabilitation. Other chapters describe the most frequent client diagnoses within the Home with a special regime, and finally the standards of social service quality are mentioned. The main research question: What are the options of using coordinated rehabilitation in a home with a special regime? Partial research question: Are employees informed about the options of coordinated rehabilitation? For the purposes of the interview I used questions focused on general identification data, questions focusing on the importance of coordinated rehabilitation, multidisciplinary team. Further, I asked about the individual components of coordinated rehabilitation, and finally, how individual planning is carried out. The result of this diploma thesis is a finding that individual components of coordinated rehabilitation are not fully mutually connected. It would be useful to provide employees with suitable courses to complete their missing knowledge. This diploma thesis might serve as a tool for improving care in the Home with a special regime, it might contribute to the Home with a special regime´s employer awareness of coordnated rehabilitiation, and connect it with practice.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Alzheimer s disease (AD) is a progressive neurodegenerative disorder, mainly characterized by memory and intellectual capacity loss. AD is characterized by deposition of amyloid-β peptide, neurofibrillary tangles, neuroinflammation, energy metabolism impairment, oxidative stress and synaptic dysfunction and loss. Its multiple pathological pathways contribute to the difficulty of AD treatment and prevention. Thus, the development of new therapies for AD curing or treatment is a challenge. The purpose of this study was to indicate an organoselenium moiety, p,p -dimethoxyl-diphenyl diselenide [(MeOPhSe)2], as a promising alternative for the treatment and prevention of sporadic dementia of Alzheimer-type (SDAT), using an experimental model of dementia induced by intracerebroventricular (i.c.v.) injection of streptozotocin (STZ) in rodents. Initially, it was investigated the prophylactic action of (MeOPhSe)2. For this, mice were treated with (MeOPhSe)2 (25 mg/kg, by gavage) and STZ (2μl of 2,5mg/ml solution; i.c.v.) or vehicles, and 48h after that, the treatment was repeated. The tasks of step-down-type passive-avoidance (SDPA), Y-maze and Morris water-maze (MWM), that followed this treatment, showed that (MeOPhSe)2 protected against the impairment in learning and memory caused by i.c.v. injection of STZ in mice. (MeOPhSe)2 protected against the increase in reactive species (RS) and the reduction of glutathione (GSH) levels, as well as modulated the antioxidant enzymes. (MeOPhSe)2 inhibited the acetylcholinesterase (AChE) activity, which was increased by STZ. Subsequently, it was investigated the effectiveness of (MeOPhSe)2 in reversing the cognitive impairment and neuronal damage induced by STZ. Therefore, rats were injected with STZ (1.0 mg/8μl; 4μl/ventricle) twice, 48h apart. After 21 days of STZ injection, regular diet fed rats were supplemented with 10ppm of (MeOPhSe)2 during 30 days. At the end of this period, it was observed that (MeOPhSe)2 dietary supplementation reversed STZ-induced memory impairment in MWM, SDPA and object recognition tasks. The results on SDPA and object recognition tasks demonstrated that the (MeOPhSe)2 improved memory in rats per se. STZ enhanced the RS and protein nitration levels in cortex and decreased GSH levels on hippocampus of rats, (MeOPhSe)2 reversed these alterations. (MeOPhSe)2 normalized AChE activity (which was enhanced by STZ) in both cortex and hippocampus, but did not reverse the deficit in cerebral glucose metabolism (ATP turnover was decrease by STZ). (MeOPhSe)2 was effective in reducing STZ-induced neuronal (apoptosis) loss. Moreover, (MeOPhSe)2 suppressed neuroinflammation induced by STZ in the rats hippocampus. The organoselenium inhibited activation of microglia and astrogliosis. Based on these results, it was concluded that: 1) (MeOPhSe)2 protected and reversed the cognitive abilities decline; 2) the mechanisms involved in the neuroprotective effect of (MeOPhSe)2 are: antioxidant, AChE inhibitor; inflammation suppressor; 3) (MeOPhSe)2 did not alter the energy metabolism; and 4) (MeOPhSe)2 reduced the neuronal death. Therefore, the present study demonstrated that (MeOPhSe)2 is a promising alternative for the drug studies for treatment of cognitive disorders such as SDAT. / A doença de Alzheimer (DA) é uma síndrome neurodegenerativa progressiva caracterizada principalmente por uma perda da memória e da capacidade intelectual. Ela é caracterizada pelo depósito de fragmentos β-amilóides; emaranhados neurofibrilares; neuroinflamação; déficit do metabolismo energético; estresse oxidativo e deficiência da neurotransmissão. As múltiplas vias patológicas da DA dificultam sua prevenção e tratamento. Logo, o desenvolvimento de novas terapias para a DA é um desafio. Por esta razão, este trabalho procurou apontar uma nova molécula orgânica contendo selênio, o disseleneto de p-metoxi fenila [(MeOPhSe)2], como uma alternativa promissora para o tratamento e prevenção da demência decorrente da DA (DEDA), usando um modelo experimental de demência induzida pela injeção intracerebroventricular (i.c.v.) de estreptozotocina (ETZ) em roedores. Inicialmente, avaliou-se o efeito profilático do (MeOPhSe)2. Para tal, camundongos receberam uma dose oral do organoselênio (25mg/kg, gavage) 30 minutos antes da ETZ (2μl de uma solução 2,5mg/ml), esse procedimento foi repetido 48horas depois. Os testes da esquiva passiva, do labirinto em Y e aquático de Morris, que sucederam esse tratamento, revelaram que o (MeOPhSe)2 protegeu os camundongos do prejuízo cognitivo induzido pela ETZ. O (MeOPhSe)2 protegeu o tecido cerebral do aumento das espécies reativas (ER) e da diminuição dos níveis de glutationa (GSH) induzidos pela ETZ, assim como modulou a atividade de enzimas antioxidantes. O (MeOPhSe)2 inibiu a atividade da acetilcolinesterase (AChE), a qual foi estimulada pela ETZ. Posteriormente, investigou-se a efetividade do (MeOPhSe)2 em reverter o prejuízo cognitivo e os danos neuronais induzidos pela ETZ. Para isso, a ETZ foi injetada nos ratos (1μg/8μl, 4μl/ventrículo) em 0 e 48horas. Passados 21dias, iniciou-se uma suplementação dietética com 10ppm de (MeOPhSe)2 durante 30dias. Ao final deste período, observou-se que o (MeOPhSe)2 restaurou as habilidades cognitivas prejudicadas pela ETZ nos ratos, nos testes do labirinto aquático de Morris, esquiva passiva e reconhecimento do objeto. Os resultados referentes aos testes do reconhecimento do objeto e da esquiva passiva apontaram que o (MeOPhSe)2 melhorou per se a memória dos ratos. A ETZ aumentou os níveis de ER e de nitração de proteínas no córtex e diminuiu os níveis de GSH no hipocampo dos ratos, o (MeOPhSe)2 reverteu estas alterações. O organoselênio inibiu a atividade da AChE (aumentada pela ETZ) tanto no córtex como no hipocampo dos ratos, mas não modulou o metabolismo da glicose (ETZ diminuiu ATP-turnover). O (MeOPhSe)2 evitou a perda neuronal (apoptose) e inibiu os eventos neurodegenerativos (ativação da caspase-3) induzidos pela ETZ. O (MeOPhSe)2 suprimiu a neuroinflamação induzida pela ETZ no hipocampo dos ratos. O organoselênio inibiu a ativação das células gliais e astrócitárias. Baseado nestes resultados, conclui-se que: 1) O (MeOPhSe)2 protegeu e reverteu o declínio das habilidades cognitivas; 2) Os mecanismos envolvidos no efeito neuroprotetor do (MeOPhSe)2 são: antioxidante; inibidor da AChE; supressor da neuroinflamação; 3) O (MeOPhSe)2 não altera o metabolismo energético; e 4) O (MeOPhSe)2 reduziu a morte neuronal. Assim sendo, este trabalho demonstrou que o (MeOPhSe)2 é uma alternativa promissora para o estudo de drogas para o tratamento de desordens cognitivas como a DEDA.

Made available in DSpace on 2016-06-02T19:46:04Z (GMT). No. of bitstreams: 1 2397.pdf: 698224 bytes, checksum: 7ce1123d029ba03759695333be6642bb (MD5) Previous issue date: 2009-03-16 / Universidade Federal de Minas Gerais / Dementia involves the loss of cognitive functions, interfering with the execution of even the most basic tasks of everyday life. Family eldercare providers are central figures in guaranteeing the wellbeing of their elderly relatives who develop dementia. However, when such a person becomes dependent on other family members, in addition to an increase in the caregiver s domestic chores, the behavior problems that arise can have a negative impact when caregivers are not well prepared for this situation, making the caregivers more susceptible to depression and contributing to the deterioration of their own physical and psychological wellbeing, making the caregiving context even more difficult. The objective of this study was to analyze the effects of psycho-educational support groups (PEG) for people caring for a relative with Alzheimer s disease (AD), associated with individual, home visits that were either training-oriented (Experimental Group EG) or listening-oriented (Control Group CG) in nature, with respect to the participants perceptions of burden, depressive symptoms and quality of life. The nine participants were randomly assigned to either the EG and CG. The EG participants received encouragement and assistance in operationalizing the concepts learned in the PEG, which addressed strategies for managing stress and depression and provided training in some of the social skills that are useful in establishing new routines (praise, constructive criticism, and asking for help). The CG received visits of the same duration, bus focused only on listening. The participants were interviewed about their situation, but did not receive individual assistance in operationalizing the strategies presented in the PEG. At the end of the first phase of the intervention, each group received the other type of home-based intervention. The data were analyzed using nonparametric tests (Mann-Whitney and Wilcoxon). The psycho-educational groups were effective in transmitting information, but did not reduce the caregivers perceptions of burden, reporting of depressive symptoms or change their quality of life ratings. After the caregivers received the training-oriented home visits (TV), their perceptions of burden significantly decreased and their quality of life ratings increased. The listeningbased visits had no significant effects on the caregivers. Six months later, an analysis of the follow-up measures indicate that improvements in perceptions of burden, frequency of depressive symptoms and quality of life scores were maintained by those who experienced the PEG coupled with TV. In conclusion, it is clear that professionals who offer orientation programs for family members caring for elderly relatives who have AD should further invest in evaluating the influence of home visits that focus on assisting these caregivers in adjusting their managerial and coping strategies, so as to increase our knowledge of how to blend group-based and individual psycho-educational supports in order to maximize the effects of such interventions. / A demência se caracteriza pela perda de funções cognitivas, comprometendo a execução normal das mais simples tarefas da vida diária. Os cuidadores familiares são pessoas chaves para garantir o bem-estar do seu familiar idoso com demência. Mas, além do aumento da carga de tarefas domésticas que ocorre quando um idoso se torna dependente, os distúrbios de comportamento do idoso dementado provocam um impacto negativo na vida dos cuidadores quando estes não são bem orientados. Isso os tornam mais vulneráveis ao desenvolvimento de quadros depressivos o que contribui para a deterioração de suas próprias condições físicas e psicológicas, complicando ainda mais a tarefa de cuidar. O presente trabalho teve como objetivo analisar os efeitos de um grupo psico-educacional (GPE) para cuidadores de idosos com doença de Alzheimer (DA), associado com um atendimento domiciliar individualizado que tinha como característica uma intervenção terapêutica com o cuidador (Grupo Experimental GE) ou a simples escuta deste (Grupo Controle GC). Os efeitos investigados incluíram as percepções dos participantes de burden, sintomas de depressão e qualidade de vida. Os nove participantes foram alocados aleatoriamente no GE e GC. No GE, os participantes receberam incentivo e suporte na operacionalização de conceitos aprendidos no GPE, os quais envolveram o manejo do estresse e da depressão e o treino de algumas habilidades sociais que são úteis no estabelecimento de rotinas novas (elogios, críticas construtivas e pedir ajuda). O grupo controle recebeu visitas com a mesma duração, mas focando apenas a escuta. Os participantes responderam a um roteiro de entrevista sobre sua situação, sem receber apoio individual na operacionalização das estratégias tratadas no GPE. Após a conclusão da primeira fase da intervenção, cada grupo recebeu o outro tipo de visita domiciliar. Análises feitas com os testes não-paramétricos Mann-Whitney e Wilcoxon mostraram que os grupos terapêuticos psico-educacionais foram eficazes para passar informação aos cuidadores, mas não aliviaram a sobrecarga ou afetaram a pontuação destes cuidadores nas medidas de sintomas depressivos ou qualidade de vida. Porém, quando os cuidadores obtiveram acompanhamento terapêutico individualizado, houve uma diminuição significativa do burden e um aumento da qualidade de vida. As visitas focando a escuta não levaram a nenhuma alteração por parte dos cuidadores. Os sintomas depressivos não foram afetados imediatamente por nenhum tipo de intervenção. Seis meses após o término das intervenções, foi feita outra avaliação (follow-up) a qual mostrou que a associação entre GPE a VTD foi efetiva na manutenção das melhorias nos escores de burden, depressão e qualidade de vida. Conclui-se que os profissionais que oferecem programas de orientação a cuidadores de idosos com demência deveriam investir mais na avaliação dos efeitos de visitas domiciliaras para ajudar cuidadores no aperfeiçoamento de suas estratégias de gerenciamento e enfrentamento do estresse e da sobrecarga. Assim, poder-se-ia conhecer melhor estes efeitos da combinação de grupos psico-educativos e de visitas domiciliares individualizadas, o que por fim, poderia ajudar a maximizar o potencial de impacto das intervenções com cuidadores de idosos com DA.

Altersassoziiert entwickeln Hunde eine Erkrankung, die in vielen Aspekten der Alzheimer-Krankheit des Menschen ähnelt. Das canine kognitive Dysfunktionssyndrom äußert sich klinisch u.a. durch Desorientierung in vertrauter Umgebung, Vergessen von Kommandos und einen gestörten Schlaf-Wach-Rhythmus. Aus der Literatur ist bekannt, dass in den Gehirnen von alten Hunden regelmäßig Aβ- und selten Tauablagerungen zu beobachten sind. Allerdings erfolgte bisher kein Nachweis des hochgradig zytotoxischen und modifizierten pE3Aβ. Auch Veränderungen der mikroglialen Morphologie wurden bisher nicht beschrieben. Insgesamt lagen in dieser Studie 24 euthanasierte Rasse- und Mischlingshunde verschiedenen Alters vor. Fünf dieser Tiere besaßen ein durchschnittliches Alter von 2,1 Jahren und dienten als Kontrollgruppe. Die anderen 19 Hunde waren 8 bis 19 Jahre alt und wurden entsprechend ihrer Größe und des Gewichts in die drei Kategorien kleine (≤ 10 kg), mittelgroße (10 – 25 kg) und große Hunde (> 25 kg) unterteilt. Die Gehirne wurden aus den Schädeln präpariert und in 4 % Paraformaldehyd fixiert. Anschließend erfolgte die Präparation des frontalen und entorhinalen Kortex sowie der Hippokampusformation, die in 30%iger Saccharoselösung vitrifiziert und mittels Methylbutan bei -80 °C eingefroren wurden. Von den Regionen wurden Kryoschnitte mit einer Dicke von 40 µm angefertigt und diese anhand immunhistologischer Färbungen auf das Vorhandensein von Ablagerungen, bestehend aus den Amyloidsubtypen Aβ8-17 und pE3Aβ, sowie aus hyperphosphorylierten Tau, untersucht. Die Morphologie und das Aktivitätsstadium der Mikroglia wurden mit Antikörpern gegen Iba1 und TAL.1B5 analysiert. Zusätzlich erfolgte eine Untersuchung anhand des Filament Tracer. Stereologische Analysemethoden wurden zur Quantifizierung der Aβ-Ablagerungen und der Mikroglia angewandt. Disseminierte Plaques fanden sich bereits ab 9 Jahren. In den untersuchten Gehirnregionen von alten Hunden zeichnete sich ein progressiver Verlauf der Ablagerungen ab. Da insbesondere kleinere Hunde ein höheres Alter erreichten als mittelgroße und große Hunde konnten in dieser Kategorie vermehrt Plaques beobachtet werden. Den alten Tieren gemein war, dass in den untersuchten Gehirnregionen pE3Aβ-Plaques häufiger vorlagen als Plaques, die aus Aβ8-17 bestanden. Kleinere parenchymale und meningeale Gefäße des frontalen Kortex schienen besonders anfällig gegenüber pE3Aβ-Ablagerungen zu sein. Im entorhinalen Kortex von kleinen Hunden war die Menge an gefäßassoziierten Aβ8-17- und pE3Aβ-Ablagerungen annähernd gleich. Bei mittelgroßen und großen Hunden dominierte im entorhinalen Kortex und ventralen Hippokampus die Anzahl an gefäßassoziierten Aβ8-17-Ablagerungen. Bei kleinen Hunden existierten im ventralen Hippokampus signifikant mehr gefäßassoziierte Aβ8-17- als pE3Aβ-Ablagerungen. Hyperphosphoryliertes Tau fand sich in der Hippokampusformation von drei Hunden im Alter von 11 bzw. 15 Jahren. Der Schweregrad war unterschiedlich ausgeprägt, sodass nur ein Hund eine hochgradige Pathologie mit NFTs und neuritischen Plaques aufwies. Einhergehend mit dem Alter und einer assoziierten Proteinpathologie fanden sich Veränderungen der mikroglialen Morphologie. Neben ramifizierten Mikroglia lagen in den untersuchten Gehirnregionen aktivierte Mikroglia vor. Einige Mikroglia wiesen Zeichen einer Seneszenz auf und waren insbesondere in den Gehirnen von Hunden mit einer hochgradigen Aβ- bzw. Tau-Pathologie vorhanden. Zusammenfassend ist festzustellen, dass mit dieser Studie eine nähere Charakterisierung des caninen kognitiven Dysfunktionssyndroms erfolgte. Die Befunde sind von hoher translationaler Bedeutung und fördern die Etablierung des Hundes als natürliches Modelltier zur Untersuchung von Alterungsprozessen des Gehirns und für die Erforschung des initialen Stadiums der Alzheimer-Krankheit.:1 Einleitung 2 Literaturübersicht 2.1 Das canine kognitive Dysfunktionssyndrom 2.2 Pathogenese der Proteinablagerungen 2.2.1 Amyloid-Pathologie 2.2.2 Tau-Pathologie 2.3 Mikroglia 2.3.1 Ursprung und Formen 2.3.2 Die Rolle der Mikroglia beim Morbus Alzheimer 2.4 Assoziation des CCDS zum Morbus Alzheimer 3 Tiere, Material und Methoden 3.1 Hunde 3.2 Gehirnproben 3.2.1 Gewinnung und Kryofixierung der Gehirne 3.2.2 Makroskopische Untersuchung der Gehirne 3.2.3 Untersuchte Gehirnregionen 3.2.4 Histologische Färbungen 3.3 Immunohistochemische und Immunfluoreszenzfärbungen 3.3.1 Antikörper und Seren 3.3.2 Protokoll der ABC-Methode 3.3.3 Protokoll zur Immunfluoreszenz 3.3.4 Kontrollen 3.4 Auswertung der Färbungen 3.4.1 Deskriptive Analyse der Präparate 3.4.2 Quantitative Analyse der immunhistochemischen Befunde 3.4.3 Statistische Auswertung 4 Ergebnisse 4.1 Anamnestische Merkmale der Hunde 4.1.1 Gruppeneinteilung in Größen- und Gewichtskategorien 4.1.2 Altersverteilung 4.1.3 Symptomatik 4.2 Pathologisch-histologische Untersuchung der Gehirne 4.2.1 Altersabhängige pathologische Gehirnveränderungen 4.2.2 Anteil der grauen Substanz des frontalen Kortex 4.2.3 Pathologisch-histologische Charakterisierung ausgewählter Hirnareale 4.2.3.1 Kontrollgruppe 4.2.3.2 Gehirne der alten Hunde 4.2.3.3 Korrelation der altersassoziierten Neuropathologie mit der Größen- und Gewichtskategorie 4.2.3.4 Statistische Auswertung 4.3 Detektion, Charakterisierung und Quantifizierung des Aβ-Proteins 4.3.1 Immunhistochemische Darstellung des Aβ-Proteins 4.3.1.1 Kontrollgruppe 4.3.1.2 Altersassoziierte Verteilung der Aβ-Ablagerungen 4.3.2 Morphologie der Aβ-Ablagerungen 4.3.3 Assoziation der Aβ-Ablagerungen mit der Größen- und Gewichtskategorie 4.3.4 Histochemische Darstellung des Aβ-Proteins 4.3.5 Quantifizierung der Aβ-Ablagerungen 4.4 Immunhistologische Darstellung von Tau-Pathologien 4.4.1 Kontrollgruppe 4.4.2 Alte Hunde der Versuchsgruppen 4.5 Charakterisierung und Quantifizierung der Mikroglia 4.5.1 Darstellung der Mikroglia 4.5.1.1 Kontrollgruppe 4.5.1.2 Alte Hunde der Versuchsgruppen 4.5.2 Assoziation der mikroglialen Morphologie zu den Größen- und Gewichtskategorien der untersuchten Hunde 4.5.3 Auswertung morphologischer Parameter mit dem Filament Tracer 4.5.4 Quantifizierung der Mikroglia 4.5.4.1 Untersuchung der Anzahl der Mikroglia in Assoziation zur Neuropathologie 4.5.4.2 Untersuchung der Anzahl der Mikroglia in Assoziation zu den Größen- und Gewichtskategorien 4.5.5 Nachweis HLA DR-positiver Mikroglia 4.5.5.1 Kontrollgruppe 4.5.5.2 Alte Versuchshunde 5 Diskussion 5.1 Hundepopulation 5.2 Pathologisch-histologische Untersuchung der Gehirne 5.3 Aβ-Pathologie 5.3.1 Parenchymale Aβ-Plaques 5.3.2 Gefäßassoziiertes Aβ-Protein 5.4 Tau-Pathologie 5.4.1 Häufigkeit innerhalb der Hundepopulation und kritische Wertung der Färbemethodik 5.4.2 Zusammenhang der Tau-Pathologie mit den Aβ-Ablagerungen 5.5 Mikroglia 5.5.1 Unterschiede in der Anzahl der Mikroglia zwischen jungen und alten Hunden 5.5.2 Unterschiede der Morphologie der Mikroglia zwischen jungen und alten Hunden 5.5.3 Detektion von dystrophischen Mikroglia in den Gehirnen von alten Hunden 5.6 Schlussfolgerungen 5.7 Ausblick 6 Zusammenfassung 7 Summary 8 Literaturverzeichnis 9 Anhang 9.1 Tabellarische Übersichten zu den Studientieren 9.2 Protokoll der H&E-Färbung 9.3 Übersicht einzelner Ergebnisse des humanen Gewebes 9.4 Tabellarische Übersichten der verwendeten Materialien Abbildungsverzeichnis Tabellenverzeichnis / Dogs develop an age-associated cognitive dysfunction syndrome with several aspects resembling Alzheimer\\\''s disease. Affected animals show signs of dis-orientation in their familiar surroundings, dementia, and a disturbed circadian rhythm. The underlying neurodegenerative disease is associated with patho-logic changes in the brain including regularly deposition of β-pleated amyloid and rarely hyperphosphorylated tau accumulation. However, there have been no reports of the highly cytotoxic and modified pE3Aβ in the canine brain. Equally, altered microglial morphology has not been documented so far. For this study 24 euthanized thoroughbred dogs and mongrels of different ages were available. Five of these animals had an average age of 2.1 years and served as control group. The remaining 19 dogs were 8 to 19 years old. Accor-ding to their height and weight these dogs were divided into 3 different categories including small (≤ 10 kg), medium (11 - 25 kg) and large dogs (> 25 kg). Brains were dissected from the skulls and fixed in 4 % paraformaldehyde. Afterwards the frontal and entorhinal cortex as well as the hippocampal for-mation were isolated, vitrificated in 30 % sucrose solution and frozen to -80 °C by methylbutane. These regions were sliced into 40 µm thick sections and subsequently stained by immunohistology in order to detect deposits of Aβ8-17, pE3Aβ and hyperphosphorylated tau, respectively. Antibodies against Iba1 and TAL.1B5 were used to analyze microglial morphology and activation status. Additionally further investigations were made with the Filament Tracer of Imaris software. Stereological analysis methods served for the quantification of Aβ depositions and microglia. Disseminated Aβ plaques were detected in dogs from 9 years on. Within the examined brain regions of elderly dogs a progressive course of Aβ depositions was observed. Especially small dogs had a longer lifespan than medium and large dogs with the result that more plaques were deposited in the brains of small dogs. Elderly dogs had in common that pE3Aβ-plaques where more often located in the examined brain regions than plaques containing Aβ8-17. Minor parenchymal and meningeal vessels seemed to be susceptible especially to pE3Aβ depositions. The amount of vessel-associated Aβ8-17 and pE3Aβ in the entorhinal cortex of small dogs was almost equal. Within the entorhinal cortex of medium and large dogs the amount of vessel-associated Aβ8-17 predominated. The ventral hippocampus of small dogs showed significantly more vessel-associated Aβ8-17 than pE3Aβ depositions. Hyperphosphorylated tau was present in the hippocampal formations of 3 dogs with an age of 11 and 15 years, respectively. The degree of severity varied with the result that only one dog showed a high-grade pathology with development of NFTs and neuritic plaques. Accompanied by age and associated protein pathology altered microglial morphology was detected. Alongside with ramified microglia, activated cells were identified in the examined brain regions. Several microglia showed signs of senescence and were present in the brains of dogs with severe Aβ and tau pathology. Summarizing, this study facilitated a further characterization of the canine cognitive dysfunction syndrome. The results are of highly translational importance and encourage the establishment of the dog as a natural animal model for studying age-associated processes and the initial stage of Alzheimer’s disease.:1 Einleitung 2 Literaturübersicht 2.1 Das canine kognitive Dysfunktionssyndrom 2.2 Pathogenese der Proteinablagerungen 2.2.1 Amyloid-Pathologie 2.2.2 Tau-Pathologie 2.3 Mikroglia 2.3.1 Ursprung und Formen 2.3.2 Die Rolle der Mikroglia beim Morbus Alzheimer 2.4 Assoziation des CCDS zum Morbus Alzheimer 3 Tiere, Material und Methoden 3.1 Hunde 3.2 Gehirnproben 3.2.1 Gewinnung und Kryofixierung der Gehirne 3.2.2 Makroskopische Untersuchung der Gehirne 3.2.3 Untersuchte Gehirnregionen 3.2.4 Histologische Färbungen 3.3 Immunohistochemische und Immunfluoreszenzfärbungen 3.3.1 Antikörper und Seren 3.3.2 Protokoll der ABC-Methode 3.3.3 Protokoll zur Immunfluoreszenz 3.3.4 Kontrollen 3.4 Auswertung der Färbungen 3.4.1 Deskriptive Analyse der Präparate 3.4.2 Quantitative Analyse der immunhistochemischen Befunde 3.4.3 Statistische Auswertung 4 Ergebnisse 4.1 Anamnestische Merkmale der Hunde 4.1.1 Gruppeneinteilung in Größen- und Gewichtskategorien 4.1.2 Altersverteilung 4.1.3 Symptomatik 4.2 Pathologisch-histologische Untersuchung der Gehirne 4.2.1 Altersabhängige pathologische Gehirnveränderungen 4.2.2 Anteil der grauen Substanz des frontalen Kortex 4.2.3 Pathologisch-histologische Charakterisierung ausgewählter Hirnareale 4.2.3.1 Kontrollgruppe 4.2.3.2 Gehirne der alten Hunde 4.2.3.3 Korrelation der altersassoziierten Neuropathologie mit der Größen- und Gewichtskategorie 4.2.3.4 Statistische Auswertung 4.3 Detektion, Charakterisierung und Quantifizierung des Aβ-Proteins 4.3.1 Immunhistochemische Darstellung des Aβ-Proteins 4.3.1.1 Kontrollgruppe 4.3.1.2 Altersassoziierte Verteilung der Aβ-Ablagerungen 4.3.2 Morphologie der Aβ-Ablagerungen 4.3.3 Assoziation der Aβ-Ablagerungen mit der Größen- und Gewichtskategorie 4.3.4 Histochemische Darstellung des Aβ-Proteins 4.3.5 Quantifizierung der Aβ-Ablagerungen 4.4 Immunhistologische Darstellung von Tau-Pathologien 4.4.1 Kontrollgruppe 4.4.2 Alte Hunde der Versuchsgruppen 4.5 Charakterisierung und Quantifizierung der Mikroglia 4.5.1 Darstellung der Mikroglia 4.5.1.1 Kontrollgruppe 4.5.1.2 Alte Hunde der Versuchsgruppen 4.5.2 Assoziation der mikroglialen Morphologie zu den Größen- und Gewichtskategorien der untersuchten Hunde 4.5.3 Auswertung morphologischer Parameter mit dem Filament Tracer 4.5.4 Quantifizierung der Mikroglia 4.5.4.1 Untersuchung der Anzahl der Mikroglia in Assoziation zur Neuropathologie 4.5.4.2 Untersuchung der Anzahl der Mikroglia in Assoziation zu den Größen- und Gewichtskategorien 4.5.5 Nachweis HLA DR-positiver Mikroglia 4.5.5.1 Kontrollgruppe 4.5.5.2 Alte Versuchshunde 5 Diskussion 5.1 Hundepopulation 5.2 Pathologisch-histologische Untersuchung der Gehirne 5.3 Aβ-Pathologie 5.3.1 Parenchymale Aβ-Plaques 5.3.2 Gefäßassoziiertes Aβ-Protein 5.4 Tau-Pathologie 5.4.1 Häufigkeit innerhalb der Hundepopulation und kritische Wertung der Färbemethodik 5.4.2 Zusammenhang der Tau-Pathologie mit den Aβ-Ablagerungen 5.5 Mikroglia 5.5.1 Unterschiede in der Anzahl der Mikroglia zwischen jungen und alten Hunden 5.5.2 Unterschiede der Morphologie der Mikroglia zwischen jungen und alten Hunden 5.5.3 Detektion von dystrophischen Mikroglia in den Gehirnen von alten Hunden 5.6 Schlussfolgerungen 5.7 Ausblick 6 Zusammenfassung 7 Summary 8 Literaturverzeichnis 9 Anhang 9.1 Tabellarische Übersichten zu den Studientieren 9.2 Protokoll der H&E-Färbung 9.3 Übersicht einzelner Ergebnisse des humanen Gewebes 9.4 Tabellarische Übersichten der verwendeten Materialien Abbildungsverzeichnis Tabellenverzeichnis

Progression and pattern of changes in different biomarkers of Alzheimer’s disease (AD) and frontotemporal lobar degeneration (FTLD) like [18F]fluorodeoxyglucose positron emission tomography (FDG-PET) and magnetic resonance imaging (MRI) have been carefully investigated over the past decades. However, there have been substantially less studies investigating the potential of combining these imaging modalities to make use of multimodal information to further improve understanding, detection and differentiation of various dementia syndromes. Further the role of preprocessing has been rarely addressed in previous research although different preprocessing algorithms have been shown to substantially affect diagnostic accuracy of dementia. In the present work common preprocessing procedures used to scale FDG-PET data were compared to each other. Further, FDG-PET and MRI information were jointly analyzed using univariate and multivariate techniques. The results suggest a highly differential effect of different scaling procedures of FDG-PET data onto detection and differentiation of various dementia syndromes. Additionally, it has been shown that combining multimodal information does further improve automatic detection and differentiation of AD and FTLD.

Die Bildung von Amyloidablagerungen ist ein Kennzeichen der Alzheimerschen Erkrankung. Hauptbestandteil dieser senilen Plaques sind sogenannte A beta Peptide, die durch proteolytische Prozessierung aus einem Vorläufermolekül (APP) gebildet werden. Die vorliegende Arbeit beschreibt die Klonierung des Meerschweinchen - APP. Diese cDNA-Sequenz zeigt auf DNA-Ebene eine Homologie zum Human-APP von ca. 90%, auf Proteinebene beträgt die Identität ca. 97 %. Damit wird ein weiterer experimenteller Beweis für die evolutionäre Konservierung des Amyloidvorläuferproteins in Säugetieren erbracht. APP mRNA wird in Meerschweinchen-Geweben ubiquitär exprimiert. Durch alternatives Spleißen wird ein zum Human-APP im wesentlichen ähnliches Isoformenmuster gebildet: Isoformen, welche eine Proteaseinhibitordomäne enthalten, werden dominierend in peripheren Organen exprimiert, dagegen ist im Zentralnervensystem das APP 695 mit über 60 % der Gesamttranskripte die bevorzugt exprimierte Isoform. Die klonierte cDNA des Meerschweinchen-APP wurde in prokaryontischen wie auch eukaryontischen Zellsystemen exprimiert. Dabei wurde die Eignung einer Anzahl von gegen Human-APP gewonnenen Antikörpern zur Detektion des Meerschweinchen-APP und seiner Prozessierungsprodukte gezeigt. Die Expression der neuronal dominierend exprimierten Isoform APP 695 des Meerschweinchen-APP in humanen Neuroblastom-Zellen zeigte keine Unterschiede hinsichtlich der APP-Prozessierung und A beta-Bildung im direkten Vergleich zu Human-APP 695. Die proteolytische Prozessierung des Proteins wurde durch Detektion der typischen Spaltprodukte in vivo (im Liquor) als auch in einem neu etablierten in vitro-Modell primär kultivierter neuronaler Zellen untersucht. Diese Zellkulturen wurden zunächst immunhistochemisch und biochemisch charakterisiert und als "mixed brain"-Typ mit einem hohen neuronalen Anteil beschrieben. Die Prozessierung des endogenen Meerschweinchen-APP in kultivierten Zellen führt dabei zur Bildung und Akkumulation aggregationsfähiger A beta - Peptide. Zur Detektion dieser Peptide wurde ein sensitiver Nachweis durch Western-Blot etabliert. Es wird damit ein Modellsystem für in vitro-Untersuchungen vorgeschlagen, welches ein Studium der Expression und Prozessierung des Amyloidvorläuferproteins unter angenähert physiologischen Bedingungen ermöglicht. / A beta peptides, the major component of neuritic plaques found in the brains of patients with Alzheimer’s disease, are derived by proteolytic processing from a larger precursor molecule (amyloid precursor protein - APP). A combination of PCR methods was used to clone and sequence APP cDNA from guinea pig (Cavia porcellus). Guinea pig APP exhibits extensive similarities to human APP in terms of primary structure, mRNA expression of differentially spliced isoforms as shown by Northern blot and RT-PCR analysis as well as proteolytic processing to amyloidogenic A beta peptides. In contrast to rat and mouse APP, guinea pig APP - recombinantly expressed in human neuroblastoma-cells - was processed indistinguishable from human APP thus excluding intrinsic sequence-specific factors influencing processing. Further studies were performed using newly established primary cell cultures of guinea pig neurons. Refined methods have been used to detect and characterize major proteolytic processing products of APP in vitro and in vivo. In conclusion, guinea pigs provide a model to study expression and processing of APP that closely resembles the physiological situation in humans and should, therefore, be important in elucidating potential strategies to prevent amyloid formation in Alzheimers Disease.

Alzheimers sjukdom är en smygande neurodegenerativ demenssjukdom som främst drabbar äldre och som karakteriseras av uppkomsten av amyloidplack och neurofibriller i hjärnan. De vanligaste symptomen är demens, kognitiva problem, inbillningar och aggressivitet. Alzheimer förekommer i två olika former, presenil och senil alzheimer. Den fullständiga mekanismen bakom alzheimer är ännu okänd men två proteiner, beta-amyloid och tau, anses ligga bakom orsaken till alzheimer. Ett tredje inblandat protein som man funnit via genetisk analys är apolipoprotein E. Idag är alzheimer ett växande problem, detta i takt med att världens befolkning blir allt äldre. En problematik finns idag då den symptomatiska behandling som finns mot alzheimer anses vara otillräcklig, ett botemedel eftersträvas därför. Immunterapi är ett botemedel som man i framtiden hoppas kunna erbjuda, detta i form av ett aktivt eller passivt vaccin verksamt mot beta-amyloid. Forskning för att finna ett sådant pågår därför just nu. AN-1792 (aktivt vaccin), CAD106 (aktivt vaccin), Bapineuzumab (passivt vaccin) och Solanezumab (passivt vaccin) är fyra vaccin som har tagits fram och testats på människor. I de vaccinstudier som gjorts har motgångar stötts på men även framgångar gjorts. AN-1792 är det vaccin som visat sig vara effektivast men med svåra biverkningar medan CAD106 är det vaccin som visat sig vara mindre effektivt men säkrast. Bapineuzumab och Solanezumab visade sig däremot båda två vara overksamma. Då flera av de vaccin som framställts har varit verksamma mot amyloidplack anser forskare att ett framtida vaccin mot alzheimer är möjligt att framställa. / Alzheimer’s disease is an insidious neurodegenerative dementia disease that primarily affects elderly and is characterized by the formation of amyloid plaques and neurofibrillary tangles in the brain. The most common symptoms are dementia, cognitive problems, delusions, and aggressiveness. Alzheimer’s occurs in two forms, presenile and senile Alzheimer’s disease. The complete mechanism behind Alzheimer’s is still unknown but two proteins, beta-amyloid and tau, are considered to be behind the cause of Alzheimer’s. A third protein involved that was found through genetic analysis is apolipoprotein E. Today, Alzheimer’s is a growing problem as the world’s population is getting older. A complex of problems exists as the symptomatic treatment available against Alzheimer’s is considered to be insufficient; a cure is therefore aimed at. Immunotherapy is a cure that hopes can be offered, this in the form of an active or a passive vaccine effective against beta-amyloid. Research to find such a vaccine is therefore under progress right now. AN-1792 (active vaccine), CAD106 (active vaccine), Bapineuzumab (passive vaccine), and Solanezumab (passive vaccine) are four vaccines that have been developed and tested on humans. In the vaccine studies that have been done setbacks have been encountered but also successes have been made. AN-1792 is the vaccine proved to be effective but with severe side effects while CAD106 is the vaccine proved to be less effective but safer. Both Bapineuzumab and Solanezumab showed to be ineffective. Since several of the produced vaccines have been active against amyloid plaques scientists believes that a future vaccine against Alzheimer’s disease is possible to make.

Dementia of Alzheimer‟s Type is a degenerative neurocognitive disease accounting for majority of Dementia‟s. It affects millions of people worldwide and thousands of people in South Africa. Apart from the economic burden this illness places on the country, it has detrimental effects for those who provide care for individuals with this illness, who are mostly spouses. The spousal caregivers bears great financial, social and emotional burden which worsens as the disease progresses. The aim of this study is to phenomenologically explore and describe the lived experiences of spousal caregivers in caring for the spouses with Dementia of Alzheimer‟s Type. This South African study was therefore qualitative in nature and was conducted in the province of KwaZulu- Natal, in the city of Pietermaritzburg. Eight participants were interviewed using a semi-structured questionnaire. Data was analysed using interpretative phenomenological analysis (IPA). The main findings of this study are the negative emotional affects that the caregiving role creates. Caregiver stress and strain is experienced as well as the experiences of various losses including lack of intimacy and ruined expectations for the future as the disease progresses. The caregiving role also created negative implications for the social lives of caregivers and coping mechanisms were seen to be very important. / Psychology / M.A. (Psychology)

Le paludisme, la maladie la plus dévastatrice des régions tropicales fait l’objet de nombreuses recherches ayant pour but de trouver des médicaments préventifs, du matériel de protection, de nouveaux traitements, des vaccins, …<p>Notre travail s’est inscrit dans la recherche des composés naturels actifs sur l’agent pathogène, le Plasmodium. Nos investigations phytochimiques et pharmacologiques ont porté sur Triclisia sacleuxii, une plante utilisée en médecine traditionnelle pour traiter diverses maladies dont deux parasitaires: la schistosomiase et l’ascardiose. Elle est aussi employée dans la préparation du poison de flèche. De plus, T. sacleuxii appartient à la famille des Menispermaceae, une famille riche en alcaloïdes bisbenzylisoquinoléiques (BBIQ). Ces composés ont de nombreuses propriétés biologiques dont l’activité antipaludique et trypanocide. Plusieurs autres espèces appartenant au genre Triclisia sont utilisées en médecine traditionnelle pour traiter la fièvre, le paludisme et d’autres pathologies. Ces éléments ont motivé la recherche dans cette plante des composés à activité antiplasmodiale. En effet, la plupart des composés que nous en avons isolés (p 12) sont actifs aussi bien sur la souche chloroquino-sensible (3D7) que sur la souche chloroquino-résistante (W2) que nous avons testées.<p>Deux d’entre eux ont en plus une activité plus élevée vis-à-vis de la souche choroquino-résistante.<p>Les composés actifs sur Plasmodium falciparum ont également montré une toxicité à l’égard de Trypanosoma brucei brucei, une sous-espèce apparentée à celles qui sont à la base de la maladie du sommeil en Afrique Centrale et de l’Est.<p>A part les usages mentionnés précédemment, T. sacleuxii est en plus employée comme antidote contre les morsures de serpents. Ce qui voudrait dire qu’elle pourrait renfermer des inhibiteurs d’enzymes.<p>Aussi, des BBIQ ont déjà démontré une activité inhibitrice de l’acétylcholinéstérase (AChE) et des phospholipases A2. Sur base de ces informations, nous avons assigné un troisième objectif à notre investigation qui cible l’AChE en correlation avec la maladie d’Alzheimer (MA). La MA est une pathologie neurodégénérative qui affecte en général les personnes âgées de plus de 60 ans, caractérisée entre autres par une perte progressive de la mémoire, une détérioration de plusieurs fonctions cognitives, de troubles neurologiques et du comportement, … Les différents extraits alcaloïdiques ont montré un degré d’inhibition de l’AChE élevé ( 80 - 90%) à une concentration de 100 μg/ml. Avec les composés purs, l’inhibition est très variable (30 - 90 %) suivant la structure. Enfin, nous avons effectué des investigations pour déterminer le mode d’action antiplasmodiale des BBIQ majeurs isolés de T. sacleuxii. Il apparaît que non seulement toutes les BBIQ n’agissent pas par un même mode d’action, mais aussi un même composé pourrait agir simultanément suivant deux ou plusieurs mécanismes différents.<p><p><p>Malaria, the most devastating disease in tropical areas, is currently a target of numerous researches, aiming to find preventive medicines, protective tools, new treatments and vaccines. In a search for antiplasmodial natural compounds, we have undertaken phytochemical and pharmacological investigations on Triclisia sacleuxii, used in traditional medicine to treat various ailments including two parasitological diseases; schistosomiasis and ascariasis. It is also used as an arrow poison.<p>Triclisia sacleuxii belongs to the Menispermaceae family, which is known to contain bisbenzylisoquinolines. These components have shown various biological activities among which antimalarial and trypanocidal activity. Furthermore, many Triclisia species are used in traditional medicine for treating fever and malaria along with other disorders.<p>With this background the research was set out to investigate on possible antiparasitic compounds active against Plasmodium falciparum.<p>Most of the compounds isolated in this work were active towards both chloroquine sensitive strain (3D7) and chloroquine resistant Plasmodium strain (W2). Interestingly some of them demonstrated selectivity for the resistant strain.<p>The compounds which displayed antiplasmodial activity also showed toxicity against Trypanosoma brucei brucei, a parasite related to those which cause sleeping sickness.<p>Besides Triclisia sacleuxii traditional uses mentioned above, it is also used as a snakebites antidote. This suggests that it might contain enzymes inhibitors. Additionally, in previous works, bisbenzylisoquinolines which are believed to be present in T. sacleuxii, have displayed phospholipases A2 and acetylcholinesterase inhibitory activity. On the basis of these informations, the third aim of our investigation targeted acetylcholinesterase (AChE), an enzyme involved in Alzheimer’s disease (AD). AD is a neurodegenerative disease occurring mostly in people aged beyond 60 years, characterised by a progressive loss of memory, impairement of multiple cognitive functions, neurological and behavior disorders, Our results have demonstrated that leaves, stems and roots alkaloidal fractions have high anti-AChE activity. Pure compounds exhibited a structure-dependent activity ranging from 30 up to 90% at a concentration of 100μg/ml).<p>Finally, we have undertaken an investigation on the antiplasmodial mode of action of the major alkaloids. It appears that not only the BBIQ do not act by a same mechanism, but also a single compound may act by more than one mode of action. / Doctorat en Sciences biomédicales et pharmaceutiques / info:eu-repo/semantics/nonPublished

In der Alzheimer-Forschung (engl.: Alzheimer Disease, AD) wird immenser Aufwand zur Entwicklung von den Krankheitsverlauf verändernden Medikamenten betrieben. Studien zeigten, dass die abnormale Aggregation des Tau-Proteins offenbar zum Zusammenbruch der Zellkommunikation führt. Niedermolekulare Substanzen, die die Tau-Protein-Aggregation inhibieren und sogar bereits gebildete Aggregate wieder auflösen können wurden entwickelt, sind jedoch oft aufgrund von schlechten Wasserlöslichkeiten nur unter Zusatz von Dimethylsulfoxid (DMSO) in Biotests einsetzbar. Durch das Design maßgeschneiderter Peptid-Polyethylenglykol (PEG)-Konjugate war die spezifische Bindung und anschließende Freisetzung ausgewählter potentieller anti-AD-Wirkstoffe in DMSO-freien Biotests möglich. Für den Entwurf der Wirkstoff-Transporter wurden Peptidbibliotheken mit Raman- und Fluoreszenz-Mikroskopie-basierten Methoden hinsichtlich der Anreicherung der Wirkstoffe an Peptiden mit hoher Wirkstoff-Bindekapazität getestet. Mithilfe von Matrix-unterstützter Laser-Desorption/Ionisation (MALDI)-Massenspektrometrie (MS/MS)-Fragmentierung konnten die Peptidsequenzen der positiven Treffer identifiziert werden. Die zugehörigen Konjugate wurden synthetisiert, mit den Wirkstoffen beladen und die entstehenden sehr gut wasserlöslichen Wirkstoff-Konjugat-Komplexe analysiert. Für biomedizinische Anwendungen sind kompakte und definierte Systeme von Vorteil. Zur Strukturaufklärung der Wirkstoff-Konjugat-Komplexe konnten zahlreiche Untersuchungen erfolgreich durchgeführt werden. Viele Komplexe wurden zudem in DMSO-freien Biotests der Tau-Protein-Aggregation eingesetzt. Die Bioverfügbarkeit der schwerlöslichen anti-AD-Wirkstoffe konnte durch die Solubilisierung mit maßgeschneiderten Peptid-PEG-Konjugaten enorm verbessert werden. Die auf Raman-aktive Substanzen erweiterte Screeningprozedur kann wahrscheinlich auf eine Großzahl von Wirkstoffen mit ungünstigen pharmakologischen Eigenschaften angewendet werden. / Considerable efforts are devoted in Alzheimer Disease (AD) research to develop disease modifying drugs. Various studies have demonstrated that abnormal aggregation of Tau protein probably interrupts communication between cells. Tau protein aggregation can be inhibited and even preformed aggregates can be redissolved by small-molecule compounds. Unfortunately, these molecules often can only be applied in limited biotests using dimethyl sulfoxide (DMSO) as co-solvent due to their poor water solubility and bioavailability. The solubilization of selected potential anti-AD drugs by tailored peptide-poly(ethylene glycol) (PEG) conjugates enabled the specific binding und subsequent release of these drugs in DMSO-free biotests. For the design of the drug conjugate carriers, large peptide libraries have been screened using Raman or fluorescence microscopy-based methods to follow drug enrichment on certain peptide library beads which exhibit high drug affinity. Identification of peptide sequences of positive hits was performed by Matrix-assisted Laser Desorption/Ionization (MALDI)-mass spectrometry (MS/MS) fragmentation. The corresponding conjugates were synthesized; loaded with the potential drugs and the resulting highly water-soluble drug transporter complexes were analyzed. Compact and defined complexes are desirable with regard to biomedical applications. Various studies on drug-peptide interactions, specifity of drug binding and influence of the different parts of the conjugates for drug capacities were performed successfully. Generated drug transporter complexes were finally tested in DMSO free bioassays. Depending on drug and peptide structures, the complexes could reach effects comparable to the drugs solubilized by DMSO. The bioavailability of poor water-soluble anti-AD compounds was largely improved. Presumably, the new developed Raman-screening procedure can be expanded to a great extent of compounds suffering from unfavorable pharmacological characteristics.