Glioblastoma multiforme (GBM) is a debilitating malignant brain tumor with expected patient survival of less than a year and limited responsiveness to most treatments, often requiring biopsy for diagnosis and invasive surgery for treatment. We propose a DNA computer system, consisting of input, computation, and output components, for diagnosis and treatment. The input component will detect the presence of three GBM biomarkers: vascular endothelial growth factor (VEGF), caveolin-1α (CAV), and B2 receptors. The computation component will include indicator segments for each of these genes, and ensure that output is only released if all the biomarkers are present. The output component will consist of the therapeutic agent interleukin-12 (IL-12). This study will designate four groups of animals: untreated tumor-free (control), tumor-inoculated (RG2), treated and tumor-free (DNA), and treated and tumor-inoculated (RG2/DNA). In the RG2 and RG2/DNA groups, we will inoculate adult male Fischer rats with RG2 cells into the striatum to induce tumor growth. Rats in the DNA and RG2/DNA groups will be implanted with the DNA system at the same location via recombinant adeno- associated viral vectors. The effectiveness of the DNA system will be evaluated through tumor size measurements, collected from brain slices stained with hematoxylin and eosin, and survival curve. Additionally, IL-12 localization will confirm the release of the output component. We anticipate that the DNA treatment will result in a decrease in tumor size, leading to smaller tumor size in the RG2/DNA group versus the RG2 group. The control group is expected to survive the longest, followed by the DNA group, then the RG2/DNA group, and finally the RG2 group. In the DNA group, IL-12 is expected to stay localized to the implantation site, remaining in its unreleased stem-loop form. On the other hand, it is expected to be released and active in the RG2/DNA group. This study provides a proof of concept to demonstrate the viability and effectiveness of a DNA system using VEGF, CAV, and B2 receptors as biomarkers and IL-12 as a therapeutic output component in the RG2 model. Further research may include varying several of the parameters used in this study, including amount of RG2 administered, choice of biomarkers, quantity and choice of output component, and choice of animal model. This system provides a promising and innovative new approach that is less invasive than surgery yet is still effective in diagnosing, targeting, and treating GBM.

Lafora disease (LD) is a rare yet invariably fatal form of epilepsy characterized by progressive degeneration of the central nervous and motor systems and accumulation of insoluble glucans within cells. LD results from mutation of either the phosphatase laforin, an enzyme that dephosphorylates cellular glycogen, or the E3 ubiquitin ligase malin, the binding partner of laforin. Currently, there are no therapeutic options for LD, or reported methods by which the specific activity of glucan phosphatases such as laforin can be easily measured. To facilitate our translational studies, we developed an assay with which the glucan phosphatase activity of laforin as well as emerging members of the glucan phosphatase family can be characterized. We then adapted this assay for the detection of endogenous laforin activity from human and mouse tissue. This laforin bioassay will prove useful in the detection of functional laforin in LD patient tissue following the application of therapies to LD patients. We subsequently developed an in vitro readthrough reporter system in order to assess the efficacy of aminoglycosides in the readthrough of laforin and malin nonsense mutations. We found that although several laforin and malin nonsense mutations exhibited significant drug-induced readthrough, the location of the epitope tag used to detect readthrough products dramatically affected our readthrough results. Cell lines established from LD patients with nonsense mutations are thus required to accurately assess the efficacy of aminoglycosides as a therapeutic option for LD. Using hydrogen-deuterium exchange mass spectrometry (DXMS), we then gained insight into the molecular etiology of several point mutations in laforin that cause LD. We identified a novel motif in the phosphatase domain of laforin that shares homology with glycosyl hydrolases (GH) and appears to play a role in the interaction of laforin with glucans. We studied the impact of the Y294N and P301L LD mutations within this GH motif on glucan binding. Surprisingly, these mutations did not reduce glucan binding as expected, rather enhancing the binding of laforin to glucans. These findings elucidate the mechanism by which laforin interacts with and acts upon glucan substrates, providing a target for the development of therapeutic compounds.

Approximately 22% of the United States population suffers from a chronic orofacial pain condition. One such condition is known as trigeminal neuropathic pain frequently reported as continuous aching and burning pain, often accompanied by intermittent electrical shock-like sensations. Dental procedures or trauma are known causes of peripheral trigeminal nerve injury and inflammation. Patients who have this type of facial pain also suffer from emotional distress. For these reasons, trigeminal neuropathic pain needs to be studied in more detail to improve the understanding of the etiology and maintenance of this condition, as well as to develop effective treatment strategies. The first experiment was focused on characterizing the behavioral aspects of the Trigeminal Inflammatory Compression (TIC) mouse model. The findings determined that the TIC injury model induced mechanical and cold hypersensitivity that persist at least 21 weeks. This orofacial, neuropathic pain condition was accompanied by anxiety- and depressive-like behaviors at week 8 post injury. The TIC injury mouse model’s chronicity and development of psychosocial impairments demonstrated its usefulness as a facial pain model. The second experiment used the mouse TIC injury model to test the ability of pioglitazone (PIO), a PPARγ agonist used clinically for treatment of diabetes, on alleviating trigeminal pain. A single low dose of PIO had no effect, but a higher dose attenuated facial pain. The third experiment determined that combining ineffective low doses of PIO and D-cycloserine (DCS) produced a potentiated anti-allodynic response of these drugs and attenuated the anxiety associated with the TIC injury. Ex vivo studies revealed that cortical mitochondrial dysfunction occurred after the TIC injury but could be reversed by the combination of DCS/PIO which improves mitochondrial function. Overall, the present studies determined that the novel mouse TIC injury model is a clinically relevant facial neuropathic pain model. The results suggest that PPARγ and brain mitochondria may represent new molecular targets for the treatment of trigeminal neuropathic pain. These studies support the future “repurposing” of PIO and DCS as well as the combination of the two drugs for this new use in patients with trigeminal neuropathic pain.

Thesis (M.S.P.H.)--University of Michigan, 1938.

Thesis (M.S.P.H.)--University of Michigan, 1938.

nÃo hà / IntroduÃÃo: A neurotoxicidade cumulativa à uma toxicidade que pode advir da terapia à base de oxaliplatina (OXL), que à a 3 geraÃÃo de agentes platinos com amplo espectro de atividade antitumoral, incluindo cÃncer colorretal, ovariano e pulmonar. A neurotoxicidade associada à OXL gera uma toxicidade dose-limitante, crÃnica, a neuropatia sensitiva perifÃrica (NSP). Objetivo: Investigar o envolvimento da endotelina-1, de receptores TRPV1 e NMDA e da substÃncia P envolvidos na patogÃnese da neuropatia sensitiva perifÃrica induzida pelo agente antineoplÃsico oxaliplatina. Materiais e mÃtodos: O estudo foi aprovado pelo Comità de Ãtica em Pesquisa Animal da UFC (protocolo n 75/12). Camundongos Swiss machos (20g) foram prÃ-tratados com antagonistas de receptores de endotelina-1 (Bosentana 100mg/kg, VO; BQ-123 e BQ-788 30Âl, intraplantar) e antagonistas do receptor TRPV1 (capsazepina, 5mg/kg, IP), antagonista do receptor NK-1 da Substancia P (apreptanto, 1mg/kg, IP) e antagonista de receptores NMDA (MK-801, 0,5mg/kg, IP) 30 minutos antes da administraÃÃo de OXL (1mg/kg, IV) por 4 semanas e meia. Paralelamente foram realizados testes nociceptivos para avaliar o desenvolvimento da neuropatia sensitiva perifÃrica. A hipernocicepÃÃo foi avaliada pelo teste de imersÃo da cauda (TIC) em Ãgua fria (10ÂC) ou aquecida (43ÂC) e pelo teste Von Frey (HPM). Em seguida, foi realizado imunofluorescÃncia do segmento medular e gÃnglio da raiz dorsal e RT-PCR. Resultados: Como resultados observou-se que com o prÃ-tratamento ao uso de OXL que houve atenuaÃÃo da hiperalgesia da NSP induzida por OXL. Ao realizar a administraÃÃo de antagonistas seletivos de endotelina-1 intraplantar na pata direita observou-se reduÃÃo significativa na hiperalgesia na pata direita (tratada) em comparaÃÃo à pata esquerda (controle). Ao analisar a expressÃo gÃnica para cFos, NK-1 e o receptor de endotelina B, observou-se que houve reduÃÃo significativa da expressÃo dos marcadores no grupo prÃ-tratado com Bosentana ao comparar com o grupo OXL, que demonstrou a expressÃo aumentada para esses marcadores. ConclusÃo: Conclui-se no presente estudo que hà evidÃncias do papel da endotelina-1, de receptores (TRPV1 e NMDA) e da substÃncia P na patogÃnese da NSP induzida pelo agente antineoplÃsico OXL. / Introduction: The cumulative neurotoxicity is a toxicity that can result from oxaliplatin-based therapy (OXL), which is the 3rd generation platinum agent with broad spectrum of antitumor activity, including colorectal, ovarian and lung cancer. Neurotoxicity associated with OXL generates a dose-limiting toxicity, chronic, peripheral sensory neuropathy (NSP). Objective: To investigate the involvement of endothelin-1, TRPV1 receptors and NMDA and substance P involved in the pathogenesis of peripheral sensory neuropathy induced by oxaliplatin antineoplastic agent. Methods: Male Swiss mice (20g) were pre-treated with antagonists of endothelin-1 receptors (Bosentan 100mg / kg orally; BQ-123 and BQ-788 30μl, intraplantar) and TRPV1 receptor antagonists (capsazepine, 5mg / kg , IP), antagonist of NK-1 receptor for substance P (apreptanto, 1 mg / kg, IP), and a NMDA receptor antagonist (MK-801, 0.5mg / kg, IP) 30 minutes before administration of OXL (1mg / kg, IV) for 4.5 weeks. Parallel nociceptive tests performed to assess the development of peripheral sensory neuropathy. The hyperalgesia assessed by the tail immersion test (ICT) in cold water (10 C) or warm (43 C) and test Von Frey (HPM). Then it was performed spinal segment, and the dorsal root ganglion immunofluorescence and RT-PCR the Ethics Committee approved the study for Animal Research UFC (Protocol 75/12). Results: The results observed when using the antagonists, as a pretreatment to the use of OXL there was attenuation of the induced hyperalgesia (NSP) OXL. Upon administration of selective antagonists of endothelin in the right paw was significant reduction in paw hyperalgesia in the right (treated) compared to the left paw (control). By analyzing the gene expression of cFos, NK-1 and endothelin B receptor, it was observed that there was significant reduction of expression of the markers in pre-treated bosentan group versus OXL group that showed increased expression for these markers. Conclusion: It was concluded in this study that there is evidence of the role of endothelin-1 receptors (TRPV1 and NMDA) and substance SP in the pathogenesis of NSP induced antineoplastic agent OXL.

Submitted by Maria Eneide de Souza Araujo (mearaujo@ipen.br) on 2015-10-08T14:17:04Z No. of bitstreams: 0 / Made available in DSpace on 2015-10-08T14:17:04Z (GMT). No. of bitstreams: 0 / Nas áreas médica e odontológica existem situações clínicas e cirúrgicas que podem ter, como consequência, um dano ao tecido conjuntivo nervoso, chamado de neuropatia. O objetivo deste trabalho é avaliar terapias para tratamento das neuropatias causadas por manipulação do feixe nervoso periférico, minimizando ou eliminando sintomas causados pela lesão. O estudo foi feito em 60 ratos machos Wistar, com a avaliação morfológica da degeneração Walleriana e da regeneração do tecido nervoso no 15º e 30º dia após o trauma. A lesão consistiu na exposição e compressão do nervo isquiático da pata direita do animal, através de três nós consecutivos com fio de sutura e distância entre eles de ~ 2mm. Três tratamentos sobre a lesão foram comparados: o uso do biomaterial 3-NERVE®, a laserterapia de baixa potência e a associação dos dois. Os resultados histológicos revelaram que o biomaterial aumentou o processo inflamatório, mas modulou a degeneração inicial, através do surgimento de células de neoformação, favorecendo a regeneração nervosa no decorrer dos trinta dias. A laserterapia foi um tratamento favorável para a parestesia porque modulou os danos do processo de degeneração inicial e estimulou o reparo do tecido desde os 15 primeiros dias. Ao atingir os 30 dias o tecido se apresentou organizado e com uma quantidade menor de tecido neoformado quando comparado com o uso do biomaterial. A associação das terapias associou as propriedades das duas terapias, pois modulou a inflamação inicial, propiciou o aumento do número de células de neoformação do tecido nervoso e favoreceu a regeneração dos feixes nervosos nas amostras de 15 e 30 dias. Conclui-se que o não tratamento dificulta ou impede a regeneração nervosa, pois qualquer uma das terapias citadas modula os eventos desencadeados pela lesão. A associação do uso da laserterapia com o 3-NERVE® mostrou melhores resultados. / Dissertação (Mestrado em Tecnologia Nuclear) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP

Introdução: A Neuropatia auditiva/Dessincronia auditiva (NA) é um transtorno que foi identificado há apenas 20 anos, os pacientes que possuíam este transtorno eram diagnosticados como deficientes auditivos como conseqüência da falha no diagnóstico. Com o surgimento do registro das Emissões Otoacústicas e sua presença no repertório de testes de avaliação auditiva, foi possível ao clínico fazer o diagnóstico de NA. Assim sendo, é possível que alguns indivíduos que foram diagnosticados como portadores de uma perda auditiva neurossensorial, tivessem, na verdade, NA. Objetivos: O objetivo deste estudo foi caracterizar o tipo e grau da deficiência auditiva em uma população de deficientes auditivos da cidade de São Paulo; verificar a época da suspeita pelos pais da deficiência auditiva bem como do diagnóstico audiológico nestes indivíduos; identificar os participantes cujas avaliações comportamentais são incompatíveis com as avaliações eletrofisiológicas, visando identificar casos de Neuropatia Auditiva e realizar estudo qualitativo de casos dos participantes com incompatibilidade de respostas nas avaliações comportamentais e eletrofisiológicas. Método: Foram avaliados 89 deficientes auditivos de três escolas especiais da cidade de São Paulo e 11 do setor de Audiologia Educacional da Faculdade de Medicina da Universidade de São Paulo através de alguns testes audiológicos: imitanciometria, audiometria tonal limiar (ATL), emissões otoacústicas (EOA) e potencial evocado auditivo do tronco-encefálico (PEATE). Resultados: Dos 100 participantes deste estudo, 99% apresentaram uma deficiência auditiva do tipo neurossensorial e em 50% o grau predominante da deficiência auditiva foi profundo, um deles não conseguiu realizar a ATL. A média de idade da suspeita dos pais foi de 15,52 meses e o diagnóstico clínico foi de 25,07 meses, em todos os grupos a suspeita dos pais foi anterior ao diagnóstico. Apenas um participante apresentou avaliações comportamentais incompatíveis com as eletrofisiólogicas, todavia, no decorrer da pesquisa o quadro foi modificado, sendo que a flutuação da audição foi o fator mais marcante observado. Conclusões: Os resultados sugerem que a NA é um transtorno raro mesmo na população dos deficientes auditivos e alertam para a necessidade de acompanhar longitudinalmente os casos de suspeita do transtorno / Introduction: Auditory Neuropathy/Auditory Dys-synchrony (AN) is a disorder identified only 20 years ago. Due to diagnosis failure, patients with this disorder were diagnosed as hearing impaired. AN diagnosis was made possible upon the appearance of OAE\'s recordings and its presence in hearing evaluation battery. Therefore, it is possible that some individuals who have been diagnosed as suffering from sensorioneural hearing loss had indeed AN. Purpose: The purpose of this study was to characterize the type and degree of auditory impairment in a population of the hearing impaired in the city of São Paulo; to verify when parents had the suspicious of the hearing impairment and when was the audiological diagnosis done in these individuals; to identify the subjects whose behavioral evaluations are not compatible with the electrophysiological evaluations in order to identify Auditory Neuropathy events and carry out a qualitative study of the cases of subjects with incompatibility of responses in the behavioral and electrophysiological evaluations. Method: 89 hearing impaired individuals from three school for the deaf in the city of São Paulo, and 11 from the Educational Audiology Division of the Medicine School of the University of São Paulo were evaluated in some audiological tests: imitanciometry, audiometer evaluation (AE), otoacoustic emissions (OAE) and Auditory Brainstem Response (ABR). Results: Out of the 100 subjects in this study, 99% presented a sensorioneural hearing loss, and in 50% the predominant degree of auditory loss was profound, one of the subjects was not able to perform AE. The average age of parents suspicion was 15.52 months and the clinical diagnosis was 25.07 months, in all the groups the parents suspicion was prior to the formal diagnosis. Only one subject presented behavioral evaluations incompatible with the electrophysiological, however, in the course of the research the chart was changed, whereas the hearing fluctuation was the most remarkable factor observed. Conclusions: The results suggest that AN is a rare disorder even in the hearing impaired community and alert the need of long term follow up in the cases of suspected disorder

Orientadores: Fernando Cendes, Clarissa Lin Yasuda / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-26T06:12:20Z (GMT). No. of bitstreams: 1 Volpato_Nathalia_M.pdf: 1909573 bytes, checksum: b9cd778b7a8b7b15e06efaa25bf2ff18 (MD5) Previous issue date: 2014 / Resumo: Introdução: Epilepsia é uma doença crônica que influencia negativamente na qualidade de vida (QV) dos pacientes. Existem evidências científicas abundantes que mostram os benefícios da atividade física (AF) para diferentes indivíduos e populações; no entanto há controvérsias em estudos que relacionam AF e epilepsia. Muitas pessoas ainda acreditam que a AF pode desencadear crises epilépticas, e pessoas com epilepsia tendem a estilo de vida sedentário. Portanto, o presente estudo teve como objetivo avaliar os hábitos de AF das pessoas com epilepsia de lobo temporal refratária; e analisar os benefícios de programa de exercício físico (EF) aeróbio para essa população. Metodologia: 79 pacientes com epilepsia de lobo temporal, recrutados no Hospital das Clínicas da UNICAMP; foram avaliados através dos questionários de nível de AF (IPAQ), de nível de QV (QOLIE-31), de Histórico de Atividade Física, e Percepção e Controle de Crises. Dos 79 pacientes, 40 aceitaram realizar o teste cardiopulmonar de esforço máximo e os testes de antropometria. As avaliações foram realizadas antes e após o programa de EF aeróbio. Utilizamos o software SYSTAT9¿, com teste de Wilcoxon para analisar diferenças entre variáveis contínuas entre grupos e intra grupos; teste exato de Fisher para analisar distribuição de frequências; e realizamos correlações de Pearson entre variáveis contínuas. O nível de significância adotado foi de p<0,05. Resultados: Os dados do questionário IPAQ mostraram diferença significativa na QV entre os grupos ativo (A) e inativo (B). Após o período de intervenção de EF aeróbio, o grupo treinamento (T) obteve melhora significativa da QV geral, da percepção de bem estar, da cognição e memória, da percepção de energia diária e fadiga, da percepção da função social, e da capacidade física em relação ao momento pré intervenção. Em contrapartida, o grupo controle (C), o qual foi orientado a manter suas atividades diárias normais, apresentou redução da QV geral durante o período, diminuição da percepção de bem estar, da cognição e memória, e da função social; e manteve a capacidade física. Discussão: Os resultados evidenciam a importância do EF como complemento ao tratamento convencional de pacientes com epilepsia de lobo temporal. O EF proporcionou melhora da QV e da capacidade física; promovendo assim, a melhora emocional e da saúde geral dessa população / Abstract: Introduction: Epilepsy is a chronic condition that causes negative impact in the quality of life (QOL). There are plenty of scientific studies supporting the benefits of physical activity (PA) for different groups; however, there are controversies in studies about the relationship between PA and epilepsy. Many people still believe that PA can precipitate epileptic seizures, in such a way that people with epilepsy tend to a sedentary lifestyle. Therefore, the present study evaluated the PA habits in people with temporal lobe epilepsy and the benefits of aerobic physical exercise (PE) program for this population. Methodology: 79 patients with temporal lobe epilepsy were recruited from the Clinical Hospital of UNICAMP. They were evaluated through questionnaires of PA level (IPAQ); level of QOL (QOLIE31); History of Physical Exercise; and Perception and Seizure¿s Control. Forty out of 79 patients agreed to do the cardiopulmonary maximal exercise test and the anthropometry tests. The evaluations were taken before and after the aerobic PE program. Statistical analyses were conducted using the SYSTAT9TM Software. The differences between continuous variables between the groups and intra groups were analyzed through the Wilcoxon test; the frequency distribution was analyzed through the Fisher¿s exact test; and the relations between continuous variables were analyzed through the Pearson correlation. The level of significance adopted was p <0.05. Results: The data from IPAQ showed significant differences in QOL between the active (A) and inactive (B) groups. After the intervention period of aerobic PE, the training group (T) obtained significant improvement in overall QOL, as well as in the wellbeing perceptions; in the cognition and memory; in the daily energy and fatigue; in the social function¿s perception and in the physical capacity. On the contrary, people in the control group (C), that were instructed to maintain their normal daily activities, suffered a decrease of QOL level along with wellbeing¿s perception; cognition and memory; social function; and maintained their physical capacity. Discussion: The results show the importance of PE as complementary treatment of patients with temporal lobe epilepsy. It showed the capacity to improve the QOL and the physical capacity, therefore, promoting emotional and the overall health improvement for this population / Mestrado / Fisiopatologia Médica / Mestra em Ciências

The purpose of this work was to examine biological mechanisms and symptom outcomes of illness uncertainty and psychological stress in Parkinson’s disease (PD). Parkinson’s disease is a chronic, progressive neurodegenerative disorder characterized by complex symptoms that fluctuate in onset, severity, level of disability, and responsiveness to treatment. In addition to characteristic motor symptoms of tremor, rigidity, bradykinesia, and postural instability, a considerable number of individuals with PD also experience debilitating pain, fatigue, and medication-induced motor complications of dyskinesia, dystonia, and on-off phenomena. The unpredictable nature of PD symptoms and motor complications coupled with the inability to halt or slow disease progression may result in uncertainty and psychological stress. Evidence is lacking regarding biological mechanisms and symptom outcomes of uncertainty and psychological stress in PD. As such, 80 men and women diagnosed with PD after the age of 49 were recruited to participate in this study. Data specific to characteristics that may contribute to uncertainty and psychobehavioral measures of uncertainty, appraisal, psychological stress, and symptom outcomes of motor symptoms, pain, and fatigue were collected. Biological measures of neuropeptide Y (NPY) and cytokines were obtained. The results revealed that participants perceived a moderate level of illness uncertainty. Uncertainty correlated significantly with motor symptoms, pain severity, and pain interference and predicted more severe pain severity and pain interference. Psychological stress correlated significantly with motor symptoms, pain severity, pain interference, and fatigue and predicted more severe symptoms across all outcomes. NPY was positively correlated with threat appraisals and psychological stress. Cytokines were below the level of detection in this sample, and not used beyond descriptive analyses. In summary, this study found uncertainty and psychological stress contributed to more severe symptom outcomes in PD. This knowledge may be used to guide future studies aimed at further elucidating biobehavioral symptom and health outcomes of uncertainty and psychological stress in PD. It will also facilitate the development of interventions specifically targeted to uncertainty and psychological stress for the ultimate purpose of improving symptom management, health outcomes, and disease progression in PD.