Resumo: O objetivo geral deste estudo foi avaliar a presença, a sintomatologia predominante e as fases do estresse, identificar estratégias de enfrentamento de problemas utilizadas por mães de crianças cardiopatas submetidas a cirurgia cardíaca, e os objetivos específicos associar as variáveis tais como estratégia de enfrentamento e cardiopatia, fase de estresse e cardiopatia, fase de estresse e faixa etária da mãe, estratégia de enfrentamento e idade da mãe, fase de estresse e estratégia de enfrentamento, classe sócio-econômica e estratégia de enfrentamento. A amostra da pesquisa constituiu-se de 60 mães de crianças cardiopatas. Foram incluídas no estudo somente as mães biológicas, as quais vivenciavam a experiência da primeira cirurgia cardíaca do filho logo após a revelação do diagnóstico. Foi um estudo epidemiológico de prevalência do tipo transversal. Os resultados indicaram que a maioria das mães apresentou estresse na fase de resistênci , fase caracterizada por demasiado estresse e vulnerabilidade a doenças, com predominância de sintomas psicológicos, a estratégia de enfrentamento prioritária foi a busca de práticas religiosas. Para a análise estatística foi utilizado o teste Qui-quadrado, que mostrou não haver associação entre as variáveis tipo de cardiopatia com as estratégias de enfrentamento P= 0,840, cardiopatia com a fase de estresse P= 0,675, fase de estresse com estratégia de enfrentamento P= 0,375, fase de estresse com faixa etária, estratégia de enfrentamento com faixa etária e classe social com estratégia de enfrentamento P= 0,444, pois os valores de P foram > 0,05. Com base nos resultados obtidos concluiu-se que é necessário uma intervenção profissional com o objetivo de acolher, orientar e disponibilizar recursos de auxílio as mães desde o diagnóstico à alta hospitalar da criança para favorecer a um enfrentamento e adaptação... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Study general objectives: to assess stress and coping in mothers of children with congenital heart disease who had undergone surgery; and specifics objectives to associate coping style, stress, mothers age, heart disease and social class. Methods: Sixty mothers whose children had undergone surgery for congenital heart disease filled the Lipp Stress Symptoms Inventory for Adults, and the Ways of Coping Scale. Results: most mothers were in the resistance stage of stress. Psychological symptoms of stress were predominant and the most used coping strategy was religion. Chi-square test showed no association (p≤0.05) between heart disease and coping style (p = 0,840), heart disease and stress stage (p = 0,675), stress stage and coping (p = 0,375), stress and age, coping and age and social class and age and social class and coping (P = 0,444). The presence of and age stress in the resistance stage in most participants indicates the need for professional intervention from diagnosis until the discharge after surgery, to favor positive coping strategies and a better quality of life for mother and child. / Orientador: Antonio Sergio Martins / Coorientador: Ulisses Alexandre Croti / Banca: Marcos Augusto de Moraes Silva / Banca: Reinaldo Ayer de Oliveira / Mestre

FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / Os recÃm-nascidos (RN) com malformaÃÃo congÃnita (MC) requerem dos profissionais de enfermagem atenÃÃo e cuidados especÃficos e individualizados. O estudo objetivou avaliar a prevalÃncia dos recÃm-nascidos com malformaÃÃes congÃnitas em instituiÃÃes pÃblicas e a assistÃncia de enfermagem prestada a essas crianÃas internadas na Unidade Neonatal (UN) nas primeiras 24 horas de vida. Estudo descritivo, transversal, quantitativo, realizado em trÃs Unidades Neonatais de instituiÃÃes hospitalares (A, B e C) de Fortaleza, Brasil. A amostra foi composta de 159 recÃm-nascidos, sendo 75 na instituiÃÃo A; 44 na B; e 40 na C. Os dados foram coletados em 2012, de janeiro a junho em A e B, e de marÃo a agosto em C. Investigou-se prontuÃrios e documentos nas unidades referidas e, posteriormente, eles foram registrados em formulÃrios prÃprios contendo variÃveis maternas e neonatais, apÃs a aprovaÃÃo pelos ComitÃs de Ãtica. Os resultados revelaram prevalÃncia de 3,3%, 2,1% e 3,6% de RN malformados nas instituiÃÃes, respectivamente. Da amostra, 53% eram masculinos, 57% com 37 a 41,6 semanas gestacionais, 52% pesando entre 2.500g e 3999g, 66% com estatura de 39 a 49 cm, Apgar no 1Â(60%) e 5Â(79%) minutos de sete a dez. Dentre as terapias implementadas ao RN, sobressaÃram-se oxigenoterapia sob Oxi-hood (42%); Dieta zero (37%); hidrataÃÃo venosa (36%); punÃÃo de acesso venoso central para infusÃes (44%); manuseio de trÃs em trÃs horas (89%) e nÃo realizaram cirurgias no perÃodo (75%). Dentre os curativos, o local destacado foi a regiÃo sacral (54%) e a cobertura com compressa estÃril (21%). As malformaÃÃes congÃnitas diagnosticadas foram categorizadas conforme classificaÃÃo do CID â 10, prevalecendo aquelas pertencentes ao Sistema Osteomuscular (30%) e Sistema Nervoso Central (SNC) (21,1%), ressaltando o PÃ torto congÃnito, Polidactilia, Hidrocefalia e Mielomeningocele. Destacaram-se as malformaÃÃes isoladas (61%) e os registros de cuidados de enfermagem envolvendo exames (24,4%) e oxigenoterapia (16,9%). Ocorreram associaÃÃes estatisticamente significantes entre as categorias de malformaÃÃes e algumas variÃveis especÃficas: (MC do SNC) x (IG, Terapia Medicamentosa); (MC do olho, ouvido, face, pescoÃo) x (IG, Uso de drogas e Escolaridade); (MC Aparelho CirculatÃrio) x (Uso de drogas e Terapia Medicamentosa); (MC Aparelho RespiratÃrio) x (Idade materna e Uso de drogas); (Fenda labial ou palatina) x (Idade materna e Uso de drogas); (Outras MC do Aparelho Digestivo) x (Modalidade VentilatÃria, Forma de NutriÃÃo e Cirurgia); (MC dos ÃrgÃos Genitais) x (Sexo, Renda familiar e Forma de NutriÃÃo); (MC Osteomuscular) x (NÃmero de GestaÃÃes e Modalidade VentilatÃria); (Outras MC) x (IG e Peso); e (Anomalias CromossÃmicas) x (Idade materna), todos com p<0,05. Concluiu-se que ainda se faz prevalente o nÃmero de MC em RN e que a equipe de enfermagem implementa cuidados de acordo com as condiÃÃes clÃnicas, da patologia e equilÃbrio hemodinÃmico de cada RN e nÃo especificamente para cada tipo de malformaÃÃo. / The newborns (NB) with congenital malformation (CM) requires from nursing professionals the performance of specific and individualized care. This study aimed at evaluating the prevalence of newborns with congenital malformation in public institutions and the nursing care provided to these children admitted to the Neonatal Unit (NU) in the first 24 hours of life. This is a descriptive, cross-sectional and quantitative study, which was conducted in three Neonatal Units of hospitals (A, B, C) from the city of Fortaleza-CE/Brazil. The sample was composed of 159 newborns; from which 75 belong to institution A; 44 to B; and 40 to C. The data were collected in 2012, from January to June in A and B, and from March to August in C. Records and documents were investigated in the aformentioned units and, subsequently, they were recorded in the proper forms containing maternal and neonatal variables, after approval by the Ethical Committee. Results showed prevalence of 3.3%, 2.1% and 3.6% of malformed newborns in the institutions, respectively. Of the sample, 53% were male, 57% with 37 to 41,6 gestation weeks, 52% weighing between 2,500 g and 3999g, 66% with height 39-49 cm, Apgar score at 1st (60%) and 5th (79%) minutes from seven to ten. Among the implemented therapies to the NB, it should be highlighted oxygenotherapy in Oxy-hood (42%); Zero diet (37%); intravenous hydration (36%); central venous access puncture for infusions (44%); handling for every three hours (89%) and did not undergo surgery throughout the period (75%). Among the dressings, the highlighted location was the sacral region (54%) and the coverage with sterile compress (21%). The diagnosed congenital malformations were categorized according to the classification of the ICD â 10, prevailing those ones belonging to the Musculoskeletal System (30%) and to the Central Nervous System (CNS) (21.1%) highlighting the Congenital clubfoot, Polydactyly, Hydrocephalus and Myelomeningocele. It should also be highlighted the isolated malformations (61%) and the nursing care records involving examinations (24.4%) and oxygenotherapy (16.9%). There were statistically significant associations between the malformations categories and some specific variables: (CM of the CNS) x (GI, Drug Therapy) (CM of the eye, ear, face, neck) x (GI, Drug use and Schooling); (CM of the Circulatory System) x (Maternal age and Drug use) (CM of the Respiratory System) x (Drug use and Drug therapy); (Cleft-lip and palate) x (Maternal age and Drug use); (Others CM of the Digestive System) x (Ventilation modality and Nutrition and Surgery forms); (CM of the Genitalia) x (Gender, Family Income and Nutrition form); (Musculoskeletal CM) x (Number of pregnancies and Ventilation modality); (Others CM) x (GI and weight), and (Chromosomal Abnormalities) x (Maternal age), all with p <0.05. We have concluded that the number of NB with CM it is still prevalent and the nursing staff implements the healthcare according to the clinical conditions, pathology and hemodynamic balance of each NB and not specifically for each type of malformation.

FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / Objetivou-se avaliar o conhecimento, a atitude e a prÃtica dos enfermeiros atuantes na EstratÃgia SaÃde da FamÃlia (ESF) acerca do controle da sÃfilis na gestaÃÃo; associar as variÃveis explanatÃrias com o conhecimento, a atitude e a prÃtica dos enfermeiros acerca do controle da sÃfilis na gestaÃÃo e comparar o conhecimento e a atitude com a prÃtica em relaÃÃo à sÃfilis na gestaÃÃo. Trata-se de um estudo avaliativo do tipo Conhecimento, Atitude e PrÃtica (CAP) e abordagem quantitativa, realizado no perÃodo de junho a agosto de 2012, com 171 enfermeiros da ESF, utilizando-se como instrumento um questionÃrio inquÃrito CAP em relaÃÃo à sÃfilis na gestaÃÃo. Os dados foram organizados em tabelas e grÃficos, segundo a estatÃstica descritiva e inferencial utilizando-se os testes Mann-Whitney, Kruskal-Wallis, Qui-quadrado e Fisher. O estudo foi aprovado pelo COMEPE/UFC com o protocolo de n 81/12. Quanto ao perfil dos enfermeiros, verificou-se uma idade mÃdia de 37,5 anos, com 90,1% do sexo feminino, 64,3% com ensino superior em instituiÃÃes pÃblicas e 77,8% sÃo especialistas, formados hà cerca de 12 anos, atuando na ESF hà uma mÃdia de 9 anos e 53,8% tinha alguma capacitaÃÃo sobre a temÃtica. Em relaÃÃo ao conhecimento dos enfermeiros, a maioria (67,3%) foi classificada como adequado, mas ainda 32,7% de enfermeiros teve conhecimento inadequado e regular. Quanto à atitude e prÃtica, observou-se 97,1% dos participantes tinham crenÃas e opiniÃes adequadas e 94,2% as colocavam em prÃtica adequadamente. Houve uma associaÃÃo estatisticamente significativa entre a instituiÃÃo de graduaÃÃo e a atitude dos enfermeiros; a autoclassificaÃÃo positiva em relaÃÃo ao conhecimento acerca da sÃfilis na gestaÃÃo com a prÃtica adequada; o conhecimento e a prÃtica, bem como entre a atitude e a prÃtica. As principais dificuldades percebidas pelos enfermeiros no controle da sÃfilis congÃnita foram: a demora dos resultados dos exames de VDRL (45,6%); a dificuldade de convocar o(s) parceiro(s) e a sua adesÃo ao tratamento (28,1%), assim como o inÃcio tardio do prÃ-natal (19,9%). Frente ao exposto, destaca-se a importÃncia do reconhecimento da sÃfilis congÃnita como um importante problema de saÃde pÃblica pelo enfermeiro, visto que a partir de suas aÃÃes adequadas e baseadas no conhecimento tÃcnico-cientÃfico podem interferir diretamente no controle da sÃfilis congÃnita, ofertando-se uma assistÃncia prÃ-natal de qualidade, integral e humanizada.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Estudos que abordem a acurÃcia das caracterÃsticas definidoras (CD) podem contribuir para o aprimoramento do raciocÃnio clÃnico, conduzindo, por conseguinte, a formulaÃÃo de diagnÃsticos de enfermagem mais condizentes com a situaÃÃo clÃnica do paciente. Diante do exposto, o estudo teve como objetivo determinar as medidas de acurÃcia das CD do diagnÃstico de enfermagem PadrÃo respiratÃrio ineficaz (PRI) em crianÃas e adolescentes com cardiopatias congÃnitas (CC). A amostra incluiu 61 crianÃas e adolescentes com idade de 5 a 17 anos, diagnosticados com CC. Estes indivÃduos foram examinados pela pesquisadora, que realizou um exame fÃsico, enfocando a avaliaÃÃo respiratÃria. Em seguida, o mÃdico que atendia o paciente foi consultado para autorizar a realizaÃÃo dos testes de funÃÃo pulmonar (espirometria e manovacuometria). As informaÃÃes obtidas a partir do exame fÃsico e realizaÃÃo dos testes foram analisadas pela pesquisadora para determinar a presenÃa ou ausÃncia das CD de PRI, com base em um protocolo previamente estabelecido. Ressalta-se que 30 crianÃas e adolescentes nÃo receberam anuÃncia mÃdica para realizar os testes de funÃÃo pulmonar. Apesar disto, estes sujeitos foram incluÃdos na amostra para determinaÃÃo do diagnÃstico apenas com base nas CD provenientes da entrevista e do exame fÃsico. Com isto, duas subamostras foram formadas, sendo a primeira composta por 30 crianÃas e adolescentes que nÃo realizaram os referidos testes, e a segunda por 31 indivÃduos que realizaram tais exames. O conjunto de CD para cada crianÃa e adolescente foi organizado em 92 planilhas. Nestas, foi assinalada a presenÃa ou ausÃncia da CD. Cada avaliaÃÃo dos indivÃduos da segunda subamostra deu origem a duas planilhas, uma apenas com as CD provenientes da entrevista e exame clÃnico, e outra com estes mesmos dados acrescidos Ãs informaÃÃes dos testes de funÃÃo pulmonar. Tais planilhas foram submetidas a dois enfermeiros diagnosticadores que determinaram a ocorrÃncia de PRI. A anÃlise das inferÃncias e as informaÃÃes sobre a ocorrÃncia das CD possibilitaram a estimativa das medidas de acurÃcia. Para a primeira subamostra, a caracterÃstica que evidenciou melhores medidas de acurÃcia foi taquipneia. As inferÃncias realizadas para a segunda subamostra, com base nos dados clÃnicos e de entrevista, apresentaram como principais CD: ortopneia e uso da musculatura acessÃria para respirar. Quando os resultados dos testes de funÃÃo pulmonar foram acrescidos, as inferÃncias dos diagnosticadores nÃo evidenciaram medidas de acurÃcia com significÃncia estatÃstica para nenhuma das CD. Com isto, os resultados do presente estudo levantam questionamentos acerca da importÃncia dos testes de funÃÃo pulmonar para a inferÃncia do diagnÃstico PRI. Ademais, algumas caracterÃsticas nÃo mostraram legitimidade para PRI, a saber: assumir uma posiÃÃo de trÃs pontos, bradipneia, diÃmetro Ãntero-posterior aumentado, capacidade vital diminuÃda, pressÃo expiratÃria diminuÃda e ventilaÃÃo-minuto diminuÃda. Assim, acredita-se que novas pesquisas sÃo necessÃrias tanto para confirmar estes achados, como para esclarecer a relaÃÃo de tais CD com o diagnÃstico em questÃo. As medidas de acurÃcia obtidas ajudaram a identificar quais CD sÃo mais representativas de PRI. AlÃm disto, os resultados possibilitaram reconhecer quais caracterÃsticas sÃo menos utilizadas durante a inferÃncia deste diagnÃstico em crianÃas e adolescentes com cardiopatias congÃnitas. / Studies which address the accuracy of the defining characteristics (DC) may contribute to the improvement of diagnostic reasoning, leading to the formulation of nursing diagnoses which are more consistent with the clinical situation of the patient. Thus, the study aimed to estimate the measures of accuracy of the DC of the nursing diagnosis Ineffective breathing pattern (IBP) in children and adolescents with congenital heart disease (CHD). The sample included 61 children and adolescents aged 5-17 years, diagnosed with CHD. These patients were examined by the researcher, who conducted a physical examination, focusing on the respiratory evaluation. Then the doctor who attended the patient was consulted to authorize the performance of pulmonary function tests (spirometry and manovacuometry). The information obtained from physical examination and from the tests were analyzed by the researcher to determine the presence or absence of DC of IBP, based on a previously established protocol. It is highlighted that 30 children and adolescents have not received medical approval to perform the pulmonary function tests. Despite this, these patients were sampled to determine the diagnosis just based on the DC from the interview and physical examination. Thus, two subsamples were formed, the first with 30 children and adolescents who did not perform such tests, and the second with 31 individuals who carried out such tests. The set of DC for each child and teenager was organized into 92 spreadsheets. In these, it was indicated the presence or absence of the DC. Each evaluation of the patients in the second subsample resulted in two spreadsheets, one only with the DC from the interview and clinical examination, and another with these same data added information from the pulmonary function tests. These spreadsheets were submitted to two nurses diagnosticians which determined the occurrence of IBP. The analysis of the inferences and the information on the occurrence of DC allowed the estimation of measures of accuracy. For the first subsample, the DC that showed better measures of accuracy was tachypnea. The inferences made for the second subsample, based on clinical data and interviews, presented as main DC: orthopnea and use of accessory muscles to breathe. When the results of pulmonary function tests were added, the inferences of the diagnosticians showed accuracy measures without statistical significance for all DC. With this, the results of this study raise questions about the importance of pulmonary function tests for the inference of the nursing diagnosis IBP. Furthermore, some DC showed no legitimacy for IBP, as follows: assumption of three point position, bradypnea, increased anterior-posterior diameter, decreased vital capacity, decreased expiratory pressure, and decreased minute ventilation. Thus, it is believed that further research is needed to confirm these findings, as well as to clarify the relationship of such DC with the diagnosis IBP. The measures of accuracy obtained helped identify which DC is more representative of IBP. Moreover, the results allowed to recognize which features are less used during the inference of this diagnosis in children and adolescents with congenital heart disease.

A opacificação do cristalino presente ao nascimento ou nos primeiros meses de vida é denominada de Catarata Congênita (CC). As primeiras semanas de vida são importantes no desenvolvimento visual, já que há diferença significativa no desempenho visual de crianças que passaram pela cirurgia até seis semanas de vida (com melhor desempenho visual) em relação às crianças que realizaram a cirurgia após seis semanas de vida. Crianças que tiveram privação visual durante a primeira infância por catarata bilateral apresentam redução da visão espacial. O objetivo deste trabalho foi verificar o impacto na Função de Sensibilidade ao Contraste (FSC) em crianças de tratamento cirúrgico de Catarata Congênita, durante a fase de desenvolvimento visual. Foram avaliados três grupos: 11 crianças do grupo controle (média idade =6,83; DP=1,38), 9 crianças do grupo CC (média idade =6,87; DP=2,03) e 15 indivíduos adultos (média idade =27,63; DP=4,16). Foi utilizado o software Metropsis para avaliação da sensibilidade ao contraste de luminância de grade senoidal nas frequências espaciais de 0,2 cpg; 0,5 cpg; 1,0 cpg; 2,0 cpg; 4,2 cpg e 8,9 cpg e luminância média de 34,4 cd/m². Houve redução em toda a curva de sensibilidade ao contraste (SC) do grupo CC quando comparado ao grupo controle e não houve relação entre o tempo de privação e a redução da SC / The opacification of the lens present at birth is called Congenital Cataract (CC). The first weeks of life are important in visual development since there is not any significant difference in visual performance of children who had surgery within six weeks of life (with better visual performance) compared with children who underwent surgery after this period. Children who have visual deprivation during infancy due to bilateral cataracts have reduced spatial vision. The aim of this study was to assess the impact on Contrast Sensitivity Function (CSF) in children with Congenital Cataract surgery during the visual development age range. We evaluated three groups: 11 control children (mean age = 6.83, SD = 1.38), 9 children in the CC group (mean age = 6.87, SD = 2.03) and 15 adults (mean age = 27.63, SD = 4.16). Metropsis software was used to evaluate sensitivity to luminance contrast of the sinusoidal grating spatial frequencies of 0.2 cpd, 0.5 cpd, 1.0 cpd, 2.0 cpd, 4.2 cpd and 8.9 cpd with an average luminance of 34.4 cd / m². There was a reduction in the contrast sensitivity function (CSF) of the CC group compared to the control group and there was no correlation between the duration of deprivation and the impairment in the SC

O hipotireoidismo congênito (HC) é um distúrbio do metabolismo, caracterizado pela produção deficiente dos hormônios tireoidianos. A literatura ressalta que crianças podem apresentar alterações cognitivas, linguísticas e problemas comportamentais, mesmo quando o diagnóstico e o tratamento iniciaram precocemente. O estudo teve como objetivo verificar desempenho em habilidades motoras, comunicativas e cognitivas de indivíduos com hipotireoidismo congênito, tratados desde o período neonatal e comparar esses achados com seus pares sem alterações da tireoide. Participaram deste estudo 15 indivíduos com o diagnóstico de hipotireoidismo congênito do gênero feminino, com idade cronológica variando entre 36 a 71 meses, com Quociente de Inteligência superior a 70, denominado Grupo Experimental (GE); e 15 indivíduos típicos, sem alterações da tireoide, denominado Grupo Comparativo (GC). Os participantes dos dois grupos foram pareados quanto à idade cronológica e nível socioeconômico. Os instrumentos de avaliação utilizados foram: Entrevista com pais; Observação do Comportamento Comunicativo; Teste de Vocabulário por Imagem Peabody TVIP; Teste de Screening de Desenvolvimento DenverII. Foi realizada avaliação psicológica, quanto ao nível intelectual, com a aplicação da Escala de Inteligência StanfordBinet. Os resultados mostraram que o desempenho das crianças do GE quanto às habilidades comunicativas, de linguagem, cognitiva e de linguagem receptiva apresentaram diferença estatística significante quando comparado com o GC. Os achados desse trabalho confirmam a interferência do HC no desenvolvimento infantil, ressaltando que indivíduos com HC, mesmo diagnosticados e tratados precocemente, podem apresentar alterações motoras, comunicativas e cognitivas e esses prejuízos poderão afetálos no decorrer da vida, inclusive nas habilidades escolares. Reiterase que estudos longitudinais são importantes para acompanhar e prevenir essas alterações. / Congenital hypothyroidism (CH) is a metabolic disorder characterized by deficient production of thyroid hormones. The literature points out that children may have cognitive, language and behavioral problems changes even when the diagnosis and treatment started early. The study aimed to verify performance in motor, communication and cognitive abilities of individuals with congenital hypothyroidism treated from the neonatal period and to compare these findings with their peers without thyroid changes. The study included 15 subjects with a diagnosis of congenital hypothyroidism females with chronological age ranging from 36 to 71 months, with Intelligence Quotient greater than 70, called Experimental Group (GE); and 15 typical individuals without thyroid changes, called Comparative Group (GC). Participants in both groups were matched for chronological age and socioeconomic status. The assessment instruments used were: Interview with parents; Observation of Communicative Behavior; Vocabulary Test Peabody Picture TVIP; Screening Test Development DenverII. Psychological assessment was conducted, on the intellectual level, with the application of the StanfordBinet Intelligence Scale. The results showed that children\'s performance of GE regarding communication skills, language, cognitive and receptive language showed statistically significant differences when compared to the GC. The findings of this study confirm the interference of HC in child development, noting that individuals with HC, even if diagnosed and treated early, may have driven, communicative and cognitive changes and these losses may affect them throughout their lives, including in school skills. It is reiterated that longitudinal studies are important to monitor and prevent these changes.

Lymphocytic Choriomeningitis Virus (LCMV) is a common arenavirus and natural murine pathogen that causes congenital neurodevelopmental disease in humans. Exposure to the virus in utero often results in severe and permanent damage the fetal brain and eyes. While usually severe, symptoms vary from case to case. Little is known about the pathological mechanism of congenital LCMV disease. Animal models of congenital LCMV infections suggest that timing of infection during gestation may influence disease outcomes; however, time alone cannot explain all of the variation observed in humans. Another possibility is that individuals are infected with different strains of LCMV. The LCMV genome is composed of four highly conserved genes, yet even single amino acid mutations can cause the virus to exhibit very different properties in animal models. However, the role of viral strain in the context of neurodevelopment remains relatively unexplored. Here, using a rat model of congenital LCMV infection, we demonstrate that three related strains of LCMV produce different patterns of infection and disease states. Infection with the highly neurotropic E350 strain induces a disease comparable to that observed in many confirmed cases of human congenital LCMV infection. While most of these animals survive to adulthood, they suffer permanent motor and behavioral abnormalities. Postmortem analyses of infected brains suggest that this strain has a proclivity for infecting mitotically active regions of the brain, including the cerebellum, olfactory bulb, hippocampus and subventricular region. E350 is not known to induce immunosuppression and viral clearance is likely mediated by a robust T-cell response. Indeed, we find high numbers of inflammatory cells in the brains of E350 animals and elevated pro-inflammatory cytokines and chemokines. The immune response, though responsible for the clearance of the virus from the brain, is also implicated in severe and sometimes permanent brain damage. The Clone 13 strain, a strain typically associated with lymphatic tissue, readily infects the brains of developing rats. Many of these animals do not live to adulthood. Those that survive exhibit extreme stunted growth, but relatively normal neurodevelopment and little discernable neurological disease. By adulthood, the brains of these animals are comparable in size and structure to controls, despite reduced body mass. The Clone 13 infects the same brain regions as the E350, but is not cleared and remained at high titers for the duration of the study. Chronic infection is likely a consequence of the immunosuppressive effects of Clone 13 on the host immune system. The WE2.2 causes a severe disease with both neurological and systemic symptoms. These animals exhibit persistent seizure like discharges during peak infection and significant motor deficits. They all fail to thrive, losing weight shortly after infection and die invariably 9-11 days post-inoculation. The brains of WE2.2 animals exhibit widespread infection of neurons in the cerebellum, hippocampus, olfactory bulbs, and cortex. WE2.2 does not cause immunosuppression and high levels of inflammatory cells are observed in the brain. Cytokine and chemokine expression is complex, without discernible trends and variable by brain region. Finally, we looked at alpha-dystroglycan (α-DG) expression in the brain and compared it with infectivity among the strains. Alpha-dystroglycan is recognized as the principle receptor for LCMV and due to mutations in the viral glycoprotein, certain strains are more dependent on α-DG for infection. Several associations between α-DG expression and viral infectivity were observed; however α-DG expression alone could not explain all the differences in infection patterns.

Congenital heart disease is the most common form of birth defect, affecting approximately 1% of liveborn babies. Secundum atrial septal defect (ASD) is the second most common form of congenital heart disease (CHD). Most cases have no known cause. Chromosomal, syndromal and teratogenic causes account for a minority of cases. The hypothesis that mutations in the ASD genes NKX2-5 and GATA4 may cause apparently sporadic ASD was tested by sequencing them in unrelated probands with ASD. In this study, 1/102 individuals with ASD had an NKX2-5 mutation, and 1/129 had a deletion of the GATA4 gene. The cardiac transcription factor TBX20 interacts with other ASD genes but had not previously been associated with human disease. Of 352 individuals with CHD, including 175 with ASD, 2 individuals, each with a family history of CHD, had pathogenic mutations in TBX20. Phenotypes included ASD, VSD, valvular abnormalities and dilated cardiomyopathy. These studies of NKX2-5, GATA4 and TBX20 indicate that dominant ASD genes account for a small minority of cases of ASD, and emphasize the considerable genetic heterogeneity in dominant ASD (also caused by mutations in MYH6 and ACTC). A new syndrome of dominant ASD and the Marcus Gunn jaw winking phenomenon is reported. Linkage to known loci was excluded, extending this heterogeneity, but a whole genome scan did not identify a candidate locus for this disorder. Previous studies of inbred laboratory mice showed an association between patent foramen ovale (PFO) and measures of atrial septal morphology, particularly septum primum length (???flap valve length??? or FVL). In humans, PFO is associated with cryptogenic stroke and migraine, and is regarded as being in a pathological contiuum with ASD. Twelve inbred strains, including 129T2/SvEms and QSi5, were studied, with generation of [129T2/SvEms x QSi5] F1, F2 and F14 mice. Studies of atrial morphology in 3017 mice confirmed the relationship between FVL and PFO but revealed considerable complexity. An F2 mapping study identified 7 significant and 6 suggestive quantitative trait loci (QTL), affecting FVL and two other traits, foramen ovale width (FOW) and crescent width (CRW). Binary analysis of PFO supported four of these.

Understanding the molecular basis of Mendelian disorders featuring joint dislocation can enhance the knowledge of genetic or cellular pathways required in joint development, and provide candidate genes for studying related complex disorders, such as developmental dysplasia of the hip. Two strategies were employed in this project to investigate Mendelian contributors to congenital joint dislocation. The first strategy was to investigate in-depth a gene known to be associated with joint dislocation. Missense mutations or small in-frame deletions in FLNB, encoding filamin B, have previously been associated with a spectrum of osteochondrodysplasias. Screening a larger cohort established FLNB as the sole underlying disease gene for atelosteogenesis type I and III and also boomerang dysplasia, which was previously thought clinically to be allelic to AOI. Mutations in FLNB cause a large proportion of Larsen syndrome cases with phenotypes reminiscent of the early case series reported. Atypical or "recessive" Larsen syndrome may therefore be due to a different underlying genetic aberration. The disease-associated amino acid substitutions or in-frame deletion/insertions cluster to two main regions of the filamin B protein: the calponin homology 2 domain of the actin-binding domain, and repeats 13-17 of the rod domain. To analyse the functions of these regions, yeast two-hybrid analyses were performed. No interactors were identified with the calponin homology 2 domain, which suggests the amino acid substitutions may disrupt actin binding or the regulation thereof. A candidate interactor, centromere protein J, was identified that binds to repeats 13-15, and could suggest a model for aberrant cell division seen in growth plates of bones of individuals with atelosteogenesis types I and III and boomerang dysplasia. The second strategy used in this project was to investigate the genetic cause of a novel syndrome featuring joint dislocation. A neurocutaneous phenotype segregated in a consanguineous New Zealand family, and through a genetic mapping strategy, a significantly linked locus was identified at 10q23 (Z = 3.63), in which segregation of a common ancestral haplotype fits the linkage hypothesis of homozygosity by descent. Candidate gene analysis and subsequent screening identified a missense mutation 2350C>T in ALDH18A1, which predicts the substitution H784Y in the encoded protein [Delta]�-pyrroline-5-carboxylate synthase (P5CS). The known function of P5CS in proline and ornithine biosynthesis was not affected by the presence of H784Y in an indirect assay, and therefore the hypothesis proposed was that a novel, unknown moonlighting function of P5CS is perturbed causing the phenotype segregating in the family. As an initial exploration of functions of P5CS in the cell, yeast two-hybrid analysis was undertaken. This project examined the contribution of two genes, FLNB and ALDH18A1, to Mendelian congenital joint dislocations. How the cellular functions of the encoded proteins in the cytoskeleton, metabolism, or signal transduction, are critical for joint development is ill understood. Future investigations aimed at identifying candidate genes that confer susceptibility to developmental dysplasia of the hip should consider candidate genes that encode proteins related in function to the products of the FLNB and ALDH18A1 genes.

The human cytomegalovirus (HCMV) viral chemokine gene, UL146, shows a high degree of variability in clinical isolates. The UL146-produced viral chemokine, vCXCL-1, has homology to CXC chemokines and is predicted to be an immune modulator that may contribute to the pathogenesis of HCMV infections. In the analysis of clinical isolates from congenitally infected infants, we found 11 distinct vCXCL-1 clades. Although the four cysteine residues that create two disulfide bonds providing the essential structure for CXC chemokines,are conserved, the N-loop region, which is important for receptor binding and activation, was hypervariable. One clade also contained a modified glutamic acid-leucine-arginine (ELR) motif (asparagine-glycine-arginine / NGR), which regulates binding to CXCR1 and CXCR2 receptors. Based on this sequence information, we hypothesize that these proteins differentially activate neutrophils, which may have a role in HCMV pathogenesis. To address these functional differences, we produced representative vCXCL-1 proteins from each of the 11 clades using a baculovirus protein expression system. Using competition binding assays, we have examined their binding affinities to either CXCR1 or CXCR2 expressed on HEK293 cells. All vCXCL-1s bound to CXCR2 with different binding affinities. Interestingly, only three vCXCL-1s bound to CXCR1 while the others demonstrated did not. We analyzed functional differences between the vCXCL-1s in calcium mobilization, adhesion molecule induction, and chemotaxis on human peripheral blood neutrophils (PBNs). Although the binding affinities to CXCR2 and/or CXCR1 were variable, all vCXCL-1s were capable of inducing intracellular calcium mobilization in PBNs and upregulating adhesion molecules on the surface of PBNs to similar levels as human CXCL1. However, the potency of the vCXCL-1s in the chemotaxis of neutrophils varied and was affinity independent. We also examined secondary chemokine production upon vCXCL-1 treatment on neutrophil-like HL60 T2 cells using real-time PCR. The results showed CCL22 induction was affinity dependent. Taken together, these results provide insights into the potential role of vCXCL-1 in HCMV pathogenesis and how the variability in these chemokines can affect neutrophil function.