Orientadores: Elizete Aparecida Lomazi Da-Costa-Pinto, Antonio de Azevedo Barros Filho / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-17T03:05:14Z (GMT). No. of bitstreams: 1 Tamega_IzildadasEiras_D.pdf: 2217221 bytes, checksum: e9d4c3645d56bdb073deb9ab2debb4f7 (MD5) Previous issue date: 2010 / Resumo: Desvios nutricionais e disfagia são comumente relatados nas avaliações de crianças com encefalopatia crônica não progressiva. Apesar da importância do diagnóstico precoce, da estimulação neuro-psico-motora, da prevenção de co-morbidades e acompanhamento nutricional, são escassos os trabalhos longitudinais que avaliaram o crescimento em lactentes de risco para ECNP. O objetivo geral deste estudo foi acompanhar, prospectivamente, o crescimento de lactentes com antecedentes de fatores de risco para ECNP e exame neurológico alterado (N= 132). Os pacientes foram examinados aos 6, 12, 18 e 24 meses de idade, enquanto atendidos em ambulatório universitário. Foram registradas situação sócio-demográfica, condições de vida e saúde e realizado exame neurológico e fonoaudiológico no 6º e 24º meses. O grupo controle incluiu lactentes saudáveis (N= 125) acompanhados em unidade básica de saúde e de mesmas características sócio-econômicas. Os objetivos específicos foram mensurar indicadores antropométricos: peso, comprimento, perímetro cefálico e compará-los com os dados do grupo controle; analisar no grupo caso os valores de circunferência braquial, prega tricipital e os respectivos escores Z; descrever: aspectos sócio-demográficos, hábitos e condições alimentares, condições de nascimento, internações, morbidades, sintomas gastrintestinais, terapias auxiliares, uso de medicamentos e cuidados maternos; identificar a prevalência de distúrbios da deglutição, constipação intestinal e de erros alimentares e investigar a existência de associação entre distúrbios de deglutição e alterações neurológicas. Na análise estatística foram utilizados os testes Qui quadrado e exato de Fisher, o teste de Mann-Whitney e de Kruskal-Wallis. Para comparar as medidas longitudinais entre os 2 grupos foi utilizada a análise de variância para medidas repetidas, seguida do teste de comparação múltipla de Tukey e o teste de perfil por contraste. Os fatores de risco mais frequentes foram os do período perinatal, observados em 121 crianças (92%), prematuridade ocorreu em cerca da metade dos casos e esteve associada a outros fatores de risco. Apenas 7 crianças foram amamentadas após os 6 meses e em 50% dos pacientes observou-se erros alimentares e duração prolongada da alimentação. Sintomas de RGE ocorreram em 44 casos (33%), constipação em 17 (13%) e ambos em 65 casos (49%); metade das crianças seguiu irregularmente as terapias auxiliares e 78% utilizava medicamentos antirefluxo e anticonvulsivos. O grau de acometimento nos resultados dos exames neurológico e fonoaudiológico apresentou correlação positiva aos 6 e 24 m. Ao nascimento, os valores de peso, comprimento e perímetro cefálico dos pacientes encefalopatas foram significativamente inferiores aos valores das crianças saudáveis. A partir do 12º mês, a diferença estatística não se manteve, embora, no grupo caso, os dados absolutos permanecessem inferiores. Esse grupo apresentou incremento positivo ao longo do tempo, nos escores Z da circunferência braquial e prega tricipital, indicando acúmulo da massa gordurosa. A gravidade da disfagia correlacionou-se a maior comprometimento antropométrico. No grupo caso, a comparação do crescimento entre nascidos a termo e prematuros mostrou valores significativamente inferiores para os prematuros. Concluindo, os lactentes com agravo neurológico apresentaram comprometimento antropométrico significativo ao nascimento e aos 6 meses, sendo que o antecedente de prematuridade esteve associado a efeito negativo significativo no crescimento dessas crianças / Abstract: Nutritional disorders and dysphagia are frequently reported in children with non progressive chronic encephalopathy (NPCE). Despite the importance of early diagnosis, neuro-psycho-motor stimulation, prevention of co-morbidities and need of nutritional advising, longitudinal studies including infants at risk for NPCE are scarce. The objective of this study was to prospectively follow growth in infants with risk factors for NPCE and with abnormal neurological examination (N=132). Children were seen from 6 to 24 months age, in a tertiary outpatient clinic. Anthropometric data, neurological condition and speechaudiology test were recorded at 6th and 24th months of life. Control group with the same socio-economic characteristics included healthy infants (N=125) followed in a primary care health center. Specific objectives were to record anthropometric indicators: weight, length, head circumference and respective Z scores and to compare them to the control group; to analyze case group values of arm circumference, triceps skin fold and respective Z scores; to describe socio-demographic aspects, habits and nutritional conditions, birth conditions, hospital admissions, co-morbidities, gastrointestinal symptoms and use of medications. Statistical analysis used chi-square, Fisher's exact, Mann-Whitney and Kruskal-Wallis tests. Analysis of variance for repeated measurements, followed by Tukey's multiple comparison test and the contrast profile test were used to compare longitudinal measurements between both groups. Most common risk factors were those occurred in perinatal period, observed in 121 children (92%), prematurity was seen in about half of cases and was associated with other risk factors. Only 7 children were breastfed after 6 months and feeding misinterpretation were seen in 50% of patients. Gastroesophageal reflux symptoms were referred in 44 infants (33%), constipation in 17 (13%) and both in 65 (49%). Adherence to therapies with speech-therapist or physiotherapist was irregular. Neurological severity was associated with dysphagia. Dysphagia severity was also associated with greater anthropometric impairment at 6th and 24th months. At birth, NPCE patient's weight, length and head circumference were significantly lower, but the statistical difference did not remain at 12th month, although with lower absolute values in the case group. The case group showed a positive increment in arm circumference and triceps skinfold Z scores throughout time, indicating accumulation of fat mass. The severity of dysphagia correlated with more severe anthropometric impairment. preterm newborns in case group showed significantly lower growth values when compared to term infants In conclusion, infants with NPCE presented significant anthropometric impairment at birth and at 6 months, and preterm infants were significantly smaller than term patients / Doutorado / Saude da Criança e do Adolescente / Doutor em Saude da Criança e do Adolescente

Les similitudes entre les mécanismes neurotoxiques responsables des encéphalopathies spongiformes Transmissibles (EST) et des synucléinoapthies, ainsi que la présence concomitante des formes pathologiques de la protéine prion et de l’α-synucléine au sein d’une même maladie neurodégénérative sont deux observations qui nous ont conduits à étudier les relations existant potentiellement entre les altérations moléculaires de l’α-synucléine et les maladies à prion. Après avoir développé des anticorps monoclonaux en immunisant avec de l’α-synucléine recombinante humaine des souris n’exprimant pas de façon endogène cet immunogène, nous avons caractérisé les altérations moléculaires de l’α-synucléine apparaissant conjointement à une symptomatologie motrice sévère lors du vieillissement de souris transgéniques (TgM83) surexprimant l’α-synucléine humaine mutée en A53T. Les essais d’inoculation intracérébrale de souris TgM83 par différentes souches de prion ont mis en évidence que la transmission de l’encéphalopathie spongiforme bovine de type H permet de déclencher chez ces animaux une maladie à prion de façon concomitante au développement d’altérations moléculaires de l’α-synucléine. Enfin, l’importante accélération de la pathologie liée a l’α-synucléine observée chez des souris TgM83 ayant été inoculées par des tissus contenant des formes altérées de l’α-synucléine, constitue un résultat soutenant le fait que la pathologie liée a l’α-synucléine serait capable de se propager expérimentalement de proche en proche, comme la protéine prion pathologique au cours des EST / The overlap of neurotoxic mecanisms involved in prion diseases and synucleinopathies, and the concomitant detection of pathological forms of prion and α-synuclein in a same neurodegenerative disease, raise questions about the existence of potential relationship between α‐synuclein molecular alteration and prion diseases. First, we developed monoclonal antibodies by immunizing mice presenting a spontaneous deletion of the α-synuclein gene with human recombinant α‐synuclein. Then, we characterized the molecular alterations appearing jointly to clinical signs during the aging of a transgenic mouse model of synucleinopathies (TgM83), overexpressing human A53T α‐synuclein. Then, an approach routinely done in the field of prion was used to trigger a synucleinopathy alongside a prion disease. For this purpose, TgM83 mice were inoculated intracerebrally by three different prion strains : transmission of H-type bovine spongiform encephalopathy allows the onset of a prion disease concomitantly to the α‐synuclein pathology developed by the TgM83 mouse model. Finally, intracerebral inoculation of TgM83 mice with brain homogenates from symptomatic mice affected by a synucleinopathy triggers an important acceleration of the α‐synuclein pathology, resulting in the early onset of motor clinical signs associated with molecular alterations of α-synuclein. These data suggest that α-synuclein alterations can be experimentally transmitted from one mouse to another, supporting the idea that, far from being confined to the transmissible spongiform encephalopathies, the « prion-like » propagation of misfolded neuronal proteins might occur in synucleinopathies

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Transmissive spongiform encephalopathies (TSEs) are neurodegenerative diseases characterized by depositions of abnormally folded prion protein (PrPTSE ) in brain. PrPTSE is at present the only specific biochemical marker of human and animal TSEs. Diagnostic tests are based on the detection of PrPres after proteinase K digestion of brain homogenate using Western blot or on the immunohistochemistry of fixed brain tissue, which are both difficult and time consuming. In this work we focused on development of a new type of tests based on PrP detection without need of proteinase K digestion. As deposits of PrPTSE remain in the body for a long time, there is a substantial chance of them being nonenzymatically modified by glycation. The detection of glycated PrPTSE may have a potential to serve as a diagnostic marker. We prepared monoclonal antibodies specific for carboxymethyl lysine/arginine modified prion protein. Bacterially expressed and purified recombinant human prion protein (rhPrP) was modified by glyoxylic acid that introduces carboxymethyl groups on lysine and arginine residues present within the molecule of the protein. Modified rhPrP (rhPrP-CML) was used for immunization of laboratory mice and hybridoma cells were prepared. Screening of cell supernatants resulted in the selection of 4...

Transmissive spongiform encephalopathies (TSEs) are neurodegenerative diseases characterized by depositions of abnormally folded prion protein (PrPTSE ) in brain. PrPTSE is at present the only specific biochemical marker of human and animal TSEs. Diagnostic tests are based on the detection of PrPres after proteinase K digestion of brain homogenate using Western blot or on the immunohistochemistry of fixed brain tissue, which are both difficult and time consuming. In this work we focused on development of a new type of tests based on PrP detection without need of proteinase K digestion. As deposits of PrPTSE remain in the body for a long time, there is a substantial chance of them being nonenzymatically modified by glycation. The detection of glycated PrPTSE may have a potential to serve as a diagnostic marker. We prepared monoclonal antibodies specific for carboxymethyl lysine/arginine modified prion protein. Bacterially expressed and purified recombinant human prion protein (rhPrP) was modified by glyoxylic acid that introduces carboxymethyl groups on lysine and arginine residues present within the molecule of the protein. Modified rhPrP (rhPrP-CML) was used for immunization of laboratory mice and hybridoma cells were prepared. Screening of cell supernatants resulted in the selection of 4...