Zika virus (ZIKV) is a sexually transmitted viral infection most frequently transmitted by mosquitoes. The source of infectious virions in the male reproductive tract has yet to be elucidated. The goals of the studies included developing and characterizing two mouse models for reproductive transmission studies and demonstration of sexual transmission of virus via artificial insemination. The mouse strains used in the study lacked receptors to interferon molecules, key signaling proteins of the host immune response. Inflammation severity was assessed during acute disease, 5-11 days after infection using a novel histopathology grading system. ZIKV proteins and genome were initially detected in epididymal epithelial cells in males. Inflammation was first observed in the epididymis and progressed to the testicle in both AG129 and Ifnar-/- males. Infection of Ifnar-/- mice may better recapitulate Zika virus pathology in humans due to milder histopathologic lesions, the presence of histologically normal sperm in epididymal tubules, and an ability to survive the acute phase of disease. In further studies, male Ifnar-/- mice were challenged subcutaneously with ZIKV. Artificial insemination fluid derived from experimentally infected males showed positive sexual transmission at 7 days post infection (DPI) but not 35 or 70 DPI. These studies show passage of virus from epididymal flush and seminal plasma to females via insemination during acute ZIKV disease in males and provides a model for sexual transmission of ZIKV.

In recent years, Zika virus (ZIKV) has garnered worldwide attention due to its epidemic spread throughout the Americas and due to the newly recognized link between ZIKV infection and neurological diseases, including microcephaly and Guillain-Barré syndrome. ZIKV is a mosquito-borne member of the genus Flavivirus, which includes the other prominent human pathogens Japanese encephalitis virus, West Nile virus, dengue virus, and yellow fever virus. Many questions about the biology of ZIKV and how it causes disease remain unanswered. Furthermore, there currently are no vaccines or licensed antiviral drugs available to treat ZIKV infection. The goal of this study was to create new tools to aid in ZIKV research and in the creation of new therapies for ZIKV infection. To accomplish this, we created two recombinant ZIKVs–one expressing a green fluorescent protein reporter gene and the other expressing a luciferase reporter gene. These additional genes will allow us to easily visualize infected cells and to precisely track levels of viral replication over time, thereby facilitating new experimental approaches and providing a means to gain insights about ZIKV. We believe that these two new versions of ZIKV will prove to be useful tools in the urgent task of better understanding how ZIKV causes disease and its links to other complications, as well as in the process of developing and testing new treatments to combat ZIKV infection.

Zika virus (ZIKV) has emerged in many regions of the world, with infection outcomes spanning from no apparent illness to crippling nervous system disease. ZIKV and its close relatives, West Nile virus, Japanese encephalitis virus, dengue virus, and yellow fever virus are primarily transmitted by mosquitoes. Three ZIKVs were selected: MR-766 (Uganda, 1947), P6-740 (Malaysia, 1966), and PRVABC-59 (Puerto Rico, 2015), whose place of origin and time of isolation differ substantially. Stable, complementary DNA (cDNA) copies of the three ZIKV RNA genomes were cloned to examine the significance of viral and host genetic variations in directing ZIKV infection outcomes. Using a new toolbox for ZIKV genome engineering and protein analysis, combined with various cell culture and mouse infection model systems, the following were determined: (1) Genome-wide landscape of viral gene products and their related species, with several immuno-reactive gene products identified in the case of all three cloned ZIKVs. (2) Viral replicability in cultured cells, varied significantly depending on the virus strain and host cell type, with one cow cell line being resistant to ZIKV infection. (3) Virus induced neurological disease in mice, differed dramatically depending on the virus dose and strain, mouse age and strain, route of infection, and presence or absence of immune system components. Overall, the findings demonstrate the impact of the viral and host genetic backgrounds on the ability of ZIKV to replicate and cause disease. The ZIKV strain-specific characterizations and molecular instruments described will provide multiple avenues for developing and testing medical countermeasures.

The world has proven unprepared for deadly viral outbreaks. Designing antiviral drugs and strategies requires a firm understanding of the interactions taken place between the proteins of the virus and human proteins. The current computational models for predicting these interactions consider only single viruses for which extensive prior knowledge is available. The two prediction frameworks in this dissertation, DeNovo and DeNovo-Human, make it possible for the first time to predict the interactions between any viral protein and human proteins. They further helped to answer critical questions about the Zika virus. DeNovo utilizes concepts from virology, bioinformatics, and machine learning to make predictions for novel viruses possible. It pools protein-protein interactions (PPIs) from different viruses sharing the same host. It further introduces taxonomic partitioning to make the reported performance reflect the situation of predicting for a novel virus. DeNovo avoids the expected low accuracy of such a prediction by introducing a negative sampling scheme that is based on sequence similarity. DeNovo achieved accuracy up to 81% and 86% when predicting for a new viral species and a new viral family, respectively. This result is comparable to the best achieved previously in single virus-host and intra-species PPI prediction cases. DeNovo predicts PPIs of a novel virus without requiring known PPIs for it, but with a limitation on the number of human proteins it can make predictions against. The second framework, DeNovo-Human, relaxes this limitation by forcing in-network prediction and random sampling while keeping the pooling technique of DeNovo. The accuracy and AUC are both promising ($>85%$, and $>91%$ respectively). DeNovo-Human facilitates predicting the virus-human PPI network. To demonstrate how the two frameworks can enrich our knowledge about virus behavior, I use them to answer interesting questions about the Zika virus. The research questions examine how the Zika virus enters human cells, fights the innate immune system, and causes microcephaly. The answers obtained are well supported by recently published Zika virus studies. / Ph. D.

Introdução: A microcefalia é um sinal clínico associado à heterogeneidade etiológica. As principais causas de microcefalia são as infecções congênitas e as anomalias congênitas. Em 2015, após o surto do zika vírus (ZIKV) e o aumento de casos de microcefalia, o Ministério da Saúde (MS) instituiu notificação compulsória para os recém-nascidos (RN) com microcefalia e/ou alterações do Sistema Nervoso Central (SNC). No Brasil, a distribuição geográfica do ZIKV ocorreu de maneira diferenciada nas regiões norte e sul, sendo que no extremo sul do país houve um menor número de infecções por esse vírus. Assim, o estado do Rio Grande do Sul (RS) tornou-se um local importante para a avaliação sistemática das causas de microcefalia neste país, independente da presença de transmissão continuada do ZIKV. Objetivos: Avaliar e descrever as causas de microcefalia dos RN notificados por microcefalia e/ou alterações do SNC no RS. Métodos: Estudo descritivo dos 162 RN com microcefalia notificada no período de dezembro de 2015 a dezembro de 2016. Destes, 99 casos foram avaliados de forma retrospectiva com base em revisão de banco de dados, e 63 casos foram avaliados de forma prospectiva em ambulatório específico do HCPA-Brasil. As etapas propostas para a avaliação (ou informações coletadas dos bancos de dados) foram: 1) histórico da gestante; 2) exame físico do RN; 3) exames para pesquisa de infecção congênita – toxoplasmose, rubéola, ZIKV e CMV (reação de cadeia da polimerase - PCR ou sorológicos); 4) exames de imagem do SNC; 5) avaliação genética (para os casos com história familiar ou suspeita de alteração genética). As crianças foram avaliadas do nascimento até conclusão diagnóstica, seguimento perdido ou término do estudo. O período de avaliação das crianças não foi superior a quatro meses. Resultados: Noventa e cinco casos (58,6%) apresentavam microcefalia grave, resultando em uma prevalência desta complicação ao nascimento de 6.5/10.000 RN. A causa foi definida em 73 dos 162 casos. Destes eram infecções congênitas 31 casos (19.3%), síndromes genéticas 19 casos (11.7%), e malformação isolada do sistema nervoso central 20 casos (12,4%). E a causa não foi identificada em 89 (54,9%). Dos 31 casos com infecções congênitas, três (9.7%) foram diagnosticados com ZIKV, seis (19.3%) com citomegalovírus, oito (25,8%) com toxoplasmose, e 14 (45.2%) com sífilis congênita. Nenhum caso de rubeola congênita foi diagnosticado e a imunidade adquirida para rubeola das mães dos RN notificados foi de 91.6%. Destes casos 14 (45.1%) apresentaram baixo peso ao nascer e 21 (66.7%) eram pequenos para idade gestacional. A microcefalia grave foi identificada em 12 (38.7%) e 6 59.2% dos casos apresentaram alterações cerebrais, o que reforça a gravidade da ação das doenças infecciosas. Conclusão: Este é o primeiro estudo a avaliar os casos de microcefalia e/ou alterações do SNC durante o surto de ZIKV no RS. A prevalência de casos de ZIKV no RS foi inferior a estados do Nordeste do Brasil. A maioria dos casos de infecção congênita apresentaram lesões neurológicas graves, principalmente os casos de ZIKV, o que pode ocasionar atraso no desenvolvimento neurológico e sequelas nestas crianças ao longo da primeira infância. No entanto, salientamos a importância das demais infecções congênitas e causas desconhecidas associadas à microcefalia no RS, independente da presença de ZIKV. / Introduction: Microcephaly is a clinical sign associated with etiological heterogeneity. The main causes of microcephaly are congenital infections and congenital anomalies. The Ministry of Health (MOH) has instituted compulsory notification for newborns with microcephaly and / or Central Nervous System (CNS) disorders in 2015, following the zika virus (ZIKV) outbreak and the increase in cases of microcephaly. In Brazil, the geographical distribution of ZIKV occurred in a differentiated way in the northern and southern regions, and in the southernmost part of the country there were fewer infections due to this virus. Thus, the state of Rio Grande do Sul (RS) has become an important site for the systematic evaluation of the causes of microcephaly in this country, regardless of the presence of continuous transmission of ZIKV. Objectives: To evaluate and describe the causes of microcephaly of newborns notified by microcephaly and / or CNS changes in RS. Methods: Descriptive study of the 162 newborns with microcephaly reported from December 2015 to December 2016. Of these, 99 cases were retrospectively evaluated based on a database review, and 63 cases were evaluated prospectively in an outpatient clinic specific to HCPA-Brazil. The proposed steps for the evaluation (or information collected from the databases) were: 1) history of the pregnant woman; 2) physical examination of the newborn; 3) screening tests for congenital infection - toxoplasmosis, rubella, ZIKV and CMV (polymerase chain reaction - PCR or serological); 4) imaging studies of the CNS; 5) genetic evaluation (for cases with family history or suspected genetic alteration). The children were evaluated from birth to completion of diagnosis, missed follow-up or termination of the study. The evaluation of them was not more than four months. Results: Ninety-five cases (58.6%) presented severe microcephaly, resulting in a prevalence of this complication at birth of 6.5 / 10,000 newborn. A definite cause was established in 73 of the 162 causes. The leading etiology was congenital infections in 31 cases (19.3%), genetic syndromes in 19 cases (11.7%), and isolated central nervous system malformation in 20 cases (12.4%). Of the 31 cases with congenital infections, three (9.7%) were diagnosed with ZIKV, six (19.3%) with cytomegalovirus, eight (25.8%) with toxoplasmosis, and 14 (45.2%) with congenital syphilis. No case of congenital rubella was diagnosed and the acquired immunity to rubella from the mothers of the newborns was 91.6%. Of these, 14 (45.1%) had low birth 8 weight and 21 (66.7%) were small for gestational age. Severe microcephaly was identified in 12 (38.7%) and 59.2% of the cases presented cerebral alterations, which reinforces the severity of the action of infectious diseases. Conclusion: This is the first study to assess the cases of microcephaly and / or CNS changes during the outbreak of ZIKV in RS. The prevalence of ZIKV cases in RS was lower than in the northeastern states of Brazil. Most cases of congenital infection have severe neurological lesions, especially cases of ZIKV, which can cause delay in neurological development and detectable sequelae in these children throughout their first infancy. However, we emphasize the importance of other congenital infections and unknown causes associated with microcephaly in RS, regardless of the presence of ZIKV.

Introdução: A microcefalia é um sinal clínico associado à heterogeneidade etiológica. As principais causas de microcefalia são as infecções congênitas e as anomalias congênitas. Em 2015, após o surto do zika vírus (ZIKV) e o aumento de casos de microcefalia, o Ministério da Saúde (MS) instituiu notificação compulsória para os recém-nascidos (RN) com microcefalia e/ou alterações do Sistema Nervoso Central (SNC). No Brasil, a distribuição geográfica do ZIKV ocorreu de maneira diferenciada nas regiões norte e sul, sendo que no extremo sul do país houve um menor número de infecções por esse vírus. Assim, o estado do Rio Grande do Sul (RS) tornou-se um local importante para a avaliação sistemática das causas de microcefalia neste país, independente da presença de transmissão continuada do ZIKV. Objetivos: Avaliar e descrever as causas de microcefalia dos RN notificados por microcefalia e/ou alterações do SNC no RS. Métodos: Estudo descritivo dos 162 RN com microcefalia notificada no período de dezembro de 2015 a dezembro de 2016. Destes, 99 casos foram avaliados de forma retrospectiva com base em revisão de banco de dados, e 63 casos foram avaliados de forma prospectiva em ambulatório específico do HCPA-Brasil. As etapas propostas para a avaliação (ou informações coletadas dos bancos de dados) foram: 1) histórico da gestante; 2) exame físico do RN; 3) exames para pesquisa de infecção congênita – toxoplasmose, rubéola, ZIKV e CMV (reação de cadeia da polimerase - PCR ou sorológicos); 4) exames de imagem do SNC; 5) avaliação genética (para os casos com história familiar ou suspeita de alteração genética). As crianças foram avaliadas do nascimento até conclusão diagnóstica, seguimento perdido ou término do estudo. O período de avaliação das crianças não foi superior a quatro meses. Resultados: Noventa e cinco casos (58,6%) apresentavam microcefalia grave, resultando em uma prevalência desta complicação ao nascimento de 6.5/10.000 RN. A causa foi definida em 73 dos 162 casos. Destes eram infecções congênitas 31 casos (19.3%), síndromes genéticas 19 casos (11.7%), e malformação isolada do sistema nervoso central 20 casos (12,4%). E a causa não foi identificada em 89 (54,9%). Dos 31 casos com infecções congênitas, três (9.7%) foram diagnosticados com ZIKV, seis (19.3%) com citomegalovírus, oito (25,8%) com toxoplasmose, e 14 (45.2%) com sífilis congênita. Nenhum caso de rubeola congênita foi diagnosticado e a imunidade adquirida para rubeola das mães dos RN notificados foi de 91.6%. Destes casos 14 (45.1%) apresentaram baixo peso ao nascer e 21 (66.7%) eram pequenos para idade gestacional. A microcefalia grave foi identificada em 12 (38.7%) e 6 59.2% dos casos apresentaram alterações cerebrais, o que reforça a gravidade da ação das doenças infecciosas. Conclusão: Este é o primeiro estudo a avaliar os casos de microcefalia e/ou alterações do SNC durante o surto de ZIKV no RS. A prevalência de casos de ZIKV no RS foi inferior a estados do Nordeste do Brasil. A maioria dos casos de infecção congênita apresentaram lesões neurológicas graves, principalmente os casos de ZIKV, o que pode ocasionar atraso no desenvolvimento neurológico e sequelas nestas crianças ao longo da primeira infância. No entanto, salientamos a importância das demais infecções congênitas e causas desconhecidas associadas à microcefalia no RS, independente da presença de ZIKV. / Introduction: Microcephaly is a clinical sign associated with etiological heterogeneity. The main causes of microcephaly are congenital infections and congenital anomalies. The Ministry of Health (MOH) has instituted compulsory notification for newborns with microcephaly and / or Central Nervous System (CNS) disorders in 2015, following the zika virus (ZIKV) outbreak and the increase in cases of microcephaly. In Brazil, the geographical distribution of ZIKV occurred in a differentiated way in the northern and southern regions, and in the southernmost part of the country there were fewer infections due to this virus. Thus, the state of Rio Grande do Sul (RS) has become an important site for the systematic evaluation of the causes of microcephaly in this country, regardless of the presence of continuous transmission of ZIKV. Objectives: To evaluate and describe the causes of microcephaly of newborns notified by microcephaly and / or CNS changes in RS. Methods: Descriptive study of the 162 newborns with microcephaly reported from December 2015 to December 2016. Of these, 99 cases were retrospectively evaluated based on a database review, and 63 cases were evaluated prospectively in an outpatient clinic specific to HCPA-Brazil. The proposed steps for the evaluation (or information collected from the databases) were: 1) history of the pregnant woman; 2) physical examination of the newborn; 3) screening tests for congenital infection - toxoplasmosis, rubella, ZIKV and CMV (polymerase chain reaction - PCR or serological); 4) imaging studies of the CNS; 5) genetic evaluation (for cases with family history or suspected genetic alteration). The children were evaluated from birth to completion of diagnosis, missed follow-up or termination of the study. The evaluation of them was not more than four months. Results: Ninety-five cases (58.6%) presented severe microcephaly, resulting in a prevalence of this complication at birth of 6.5 / 10,000 newborn. A definite cause was established in 73 of the 162 causes. The leading etiology was congenital infections in 31 cases (19.3%), genetic syndromes in 19 cases (11.7%), and isolated central nervous system malformation in 20 cases (12.4%). Of the 31 cases with congenital infections, three (9.7%) were diagnosed with ZIKV, six (19.3%) with cytomegalovirus, eight (25.8%) with toxoplasmosis, and 14 (45.2%) with congenital syphilis. No case of congenital rubella was diagnosed and the acquired immunity to rubella from the mothers of the newborns was 91.6%. Of these, 14 (45.1%) had low birth 8 weight and 21 (66.7%) were small for gestational age. Severe microcephaly was identified in 12 (38.7%) and 59.2% of the cases presented cerebral alterations, which reinforces the severity of the action of infectious diseases. Conclusion: This is the first study to assess the cases of microcephaly and / or CNS changes during the outbreak of ZIKV in RS. The prevalence of ZIKV cases in RS was lower than in the northeastern states of Brazil. Most cases of congenital infection have severe neurological lesions, especially cases of ZIKV, which can cause delay in neurological development and detectable sequelae in these children throughout their first infancy. However, we emphasize the importance of other congenital infections and unknown causes associated with microcephaly in RS, regardless of the presence of ZIKV.

Introdução: A microcefalia é um sinal clínico associado à heterogeneidade etiológica. As principais causas de microcefalia são as infecções congênitas e as anomalias congênitas. Em 2015, após o surto do zika vírus (ZIKV) e o aumento de casos de microcefalia, o Ministério da Saúde (MS) instituiu notificação compulsória para os recém-nascidos (RN) com microcefalia e/ou alterações do Sistema Nervoso Central (SNC). No Brasil, a distribuição geográfica do ZIKV ocorreu de maneira diferenciada nas regiões norte e sul, sendo que no extremo sul do país houve um menor número de infecções por esse vírus. Assim, o estado do Rio Grande do Sul (RS) tornou-se um local importante para a avaliação sistemática das causas de microcefalia neste país, independente da presença de transmissão continuada do ZIKV. Objetivos: Avaliar e descrever as causas de microcefalia dos RN notificados por microcefalia e/ou alterações do SNC no RS. Métodos: Estudo descritivo dos 162 RN com microcefalia notificada no período de dezembro de 2015 a dezembro de 2016. Destes, 99 casos foram avaliados de forma retrospectiva com base em revisão de banco de dados, e 63 casos foram avaliados de forma prospectiva em ambulatório específico do HCPA-Brasil. As etapas propostas para a avaliação (ou informações coletadas dos bancos de dados) foram: 1) histórico da gestante; 2) exame físico do RN; 3) exames para pesquisa de infecção congênita – toxoplasmose, rubéola, ZIKV e CMV (reação de cadeia da polimerase - PCR ou sorológicos); 4) exames de imagem do SNC; 5) avaliação genética (para os casos com história familiar ou suspeita de alteração genética). As crianças foram avaliadas do nascimento até conclusão diagnóstica, seguimento perdido ou término do estudo. O período de avaliação das crianças não foi superior a quatro meses. Resultados: Noventa e cinco casos (58,6%) apresentavam microcefalia grave, resultando em uma prevalência desta complicação ao nascimento de 6.5/10.000 RN. A causa foi definida em 73 dos 162 casos. Destes eram infecções congênitas 31 casos (19.3%), síndromes genéticas 19 casos (11.7%), e malformação isolada do sistema nervoso central 20 casos (12,4%). E a causa não foi identificada em 89 (54,9%). Dos 31 casos com infecções congênitas, três (9.7%) foram diagnosticados com ZIKV, seis (19.3%) com citomegalovírus, oito (25,8%) com toxoplasmose, e 14 (45.2%) com sífilis congênita. Nenhum caso de rubeola congênita foi diagnosticado e a imunidade adquirida para rubeola das mães dos RN notificados foi de 91.6%. Destes casos 14 (45.1%) apresentaram baixo peso ao nascer e 21 (66.7%) eram pequenos para idade gestacional. A microcefalia grave foi identificada em 12 (38.7%) e 6 59.2% dos casos apresentaram alterações cerebrais, o que reforça a gravidade da ação das doenças infecciosas. Conclusão: Este é o primeiro estudo a avaliar os casos de microcefalia e/ou alterações do SNC durante o surto de ZIKV no RS. A prevalência de casos de ZIKV no RS foi inferior a estados do Nordeste do Brasil. A maioria dos casos de infecção congênita apresentaram lesões neurológicas graves, principalmente os casos de ZIKV, o que pode ocasionar atraso no desenvolvimento neurológico e sequelas nestas crianças ao longo da primeira infância. No entanto, salientamos a importância das demais infecções congênitas e causas desconhecidas associadas à microcefalia no RS, independente da presença de ZIKV. / Introduction: Microcephaly is a clinical sign associated with etiological heterogeneity. The main causes of microcephaly are congenital infections and congenital anomalies. The Ministry of Health (MOH) has instituted compulsory notification for newborns with microcephaly and / or Central Nervous System (CNS) disorders in 2015, following the zika virus (ZIKV) outbreak and the increase in cases of microcephaly. In Brazil, the geographical distribution of ZIKV occurred in a differentiated way in the northern and southern regions, and in the southernmost part of the country there were fewer infections due to this virus. Thus, the state of Rio Grande do Sul (RS) has become an important site for the systematic evaluation of the causes of microcephaly in this country, regardless of the presence of continuous transmission of ZIKV. Objectives: To evaluate and describe the causes of microcephaly of newborns notified by microcephaly and / or CNS changes in RS. Methods: Descriptive study of the 162 newborns with microcephaly reported from December 2015 to December 2016. Of these, 99 cases were retrospectively evaluated based on a database review, and 63 cases were evaluated prospectively in an outpatient clinic specific to HCPA-Brazil. The proposed steps for the evaluation (or information collected from the databases) were: 1) history of the pregnant woman; 2) physical examination of the newborn; 3) screening tests for congenital infection - toxoplasmosis, rubella, ZIKV and CMV (polymerase chain reaction - PCR or serological); 4) imaging studies of the CNS; 5) genetic evaluation (for cases with family history or suspected genetic alteration). The children were evaluated from birth to completion of diagnosis, missed follow-up or termination of the study. The evaluation of them was not more than four months. Results: Ninety-five cases (58.6%) presented severe microcephaly, resulting in a prevalence of this complication at birth of 6.5 / 10,000 newborn. A definite cause was established in 73 of the 162 causes. The leading etiology was congenital infections in 31 cases (19.3%), genetic syndromes in 19 cases (11.7%), and isolated central nervous system malformation in 20 cases (12.4%). Of the 31 cases with congenital infections, three (9.7%) were diagnosed with ZIKV, six (19.3%) with cytomegalovirus, eight (25.8%) with toxoplasmosis, and 14 (45.2%) with congenital syphilis. No case of congenital rubella was diagnosed and the acquired immunity to rubella from the mothers of the newborns was 91.6%. Of these, 14 (45.1%) had low birth 8 weight and 21 (66.7%) were small for gestational age. Severe microcephaly was identified in 12 (38.7%) and 59.2% of the cases presented cerebral alterations, which reinforces the severity of the action of infectious diseases. Conclusion: This is the first study to assess the cases of microcephaly and / or CNS changes during the outbreak of ZIKV in RS. The prevalence of ZIKV cases in RS was lower than in the northeastern states of Brazil. Most cases of congenital infection have severe neurological lesions, especially cases of ZIKV, which can cause delay in neurological development and detectable sequelae in these children throughout their first infancy. However, we emphasize the importance of other congenital infections and unknown causes associated with microcephaly in RS, regardless of the presence of ZIKV.

Introduction: At the end of 2015, zika virus became the protagonist of an epidemic of congenital anomalies never observed. The northeastern region of Brazil was the most affected. Congenital Zika Syndrome is characterized by severe microcephaly, critical brain damage, ophthalmologic, auditory, cardiac and orthopedic anomalies, as well as severe developmental delay with irritability, spasticity and convulsions. Owning to the fact that it is a new pathology, little is known about its long-term evolution, since affected children are aged around 3 years. Goal: To follow-up of a cohort of infants born with microcephaly and / or anomalies associated with congenital zika virus infection, from birth to 18 months of age, evaluating their growth, development, feeding and occurrence of associated morbidities. Methodology: This was a longitudinal, observational and descriptive study of a cohort of infants born in Sergipe during the outbreak of microcephaly and referred to two public health services. The children were followed up through 18 months of age in childcare consultations, together with expert evaluations and complementary examinations. The data was collected from August / 2017 to January / 2018 with using a research form. Statistical analyzes were carried out in R Core Team 2018 software. Results: The cohort comprised 84 children with Congenital Zika Syndrome. There was a predominance of females (53.8%) and only 9 newborns had no diagnosis of microcephaly but had other alterations compatible with Congenital Zika Syndrome. The Z scores for head circumference, weight and length remained stable over time, remaining below the expected standard for the three anthropometric indexes. The evolution of Z scores for weight / length showed a downward trend, although the average remained in the eutrophic pattern. In addition to the occurrence of other neurological impairments, such as seizures (69%), spasticity (48,8%), and irritability (64,3%), the infants presented severe developmental delays with delayed acquisition of all markers. The prevalence of exclusive breastfeeding until 6 months was law (14.3%) and a significant percentage of feeding difficulty (57,1%). This aspect reflected the delay in the introduction of complementary feeding (mean age of 7.1 months) and non-progression to the family diet in 22.6%. As to the complementary evaluation, cerebral malformations compatible with congenital Zika infection were detected in all children, ocular involvement was diagnosed in 42 infants (54.5%), and in half (50.7%) cardiac anomalies were observed. The main clinical morbidity observed was upper airways infection, followed by intestinal constipation. Conclusions: Infants with Congenital Zika Syndrome exhibited anthropometric growth impairment, as well as a severe delay in the acquisition of neuromotor development markers. It was found low prevalence of exclusive breastfeeding until 6 months, with a high frequency of feeding difficulties. There was also a significant number of infants who presented irritability, convulsion and spasticity. This study reinforces the need for specialized multiprofessional follow-up aimed at rehabilitation therapies and support to the family members involved. / Introdução: Ao final de 2015, o Zika vírus tornou-se protagonista de uma epidemia de anomalias congênitas jamais observada, sendo a região nordeste do Brasil a mais atingida. A Síndrome da Zika Congênita caracteriza-se por microcefalia com grave dano ao tecido cerebral, alterações oftalmológicas, auditivas, cardíacas e ortopédicas, além de crítico atraso do desenvolvimento, com irritabilidade, espasticidade e convulsões. Tratando-se de nova condição clínica, pouco se sabe sobre sua evolução em longo prazo, uma vez que as crianças acometidas estão com média de 3 anos de idade. Objetivo: Realizar o acompanhamento de uma coorte de bebês com Síndrome da Zika Congênita, do nascimento aos 18 meses de vida, avaliando seu crescimento, desenvolvimento, evolução da alimentação e ocorrência de morbidades. Metodologia: Trata-se de estudo longitudinal, observacional e descritivo do acompanhamento de uma coorte de lactentes nascidos em Sergipe durante o surto de microcefalia e referenciados para dois serviços públicos de saúde. As crianças foram avaliadas até os 18 meses de vida em consultas de puericultura somadas a interconsultas com especialistas e realização de exames complementares. Os dados foram coletados de agosto/2017 a janeiro/2018 através de um formulário de pesquisa. As análises estatísticas foram realizadas com o auxílio do software R Core 2018. Resultados: Compuseram a coorte 84 crianças com características clínicas da Síndrome da Zika Congênita. Houve predomínio do sexo feminino (53,8%) e somente 9 recém-nascidos não tiveram diagnóstico de microcefalia, mas apresentavam outras alterações compatíveis com a Síndrome da Zika Congênita. Os escores Z para perímetro cefálico (PC), peso e comprimento apresentaram pouca variação ao longo do tempo. As médias de escore Z na primeira e última consulta foram as seguintes: PC (-6,0; -5,9); Peso (-1,9; -1,6) e Comprimento (-2,5; -1,7). A evolução dos escores Z para peso/comprimento, revelou tendência de queda, apesar da média ter se mantido no padrão de eutrofia. As crianças manifestaram grave atraso do desenvolvimento com retardo na aquisição de todos os marcos pesquisados, além da ocorrência de outros comprometimentos neurológicos tais quais convulsão (69%), espasticidade (48,8%) e irritabilidade (64,3%). Encontrou-se baixa prevalência de aleitamento materno exclusivo até os 6 meses (14,3%) e percentual significativo de relatos de dificuldade alimentar (57,1%), aspecto que refletiu no atraso da introdução da alimentação complementar (idade média de 7,1 meses) e na não progressão para a alimentação da família em 22,6%. Quanto aos exames e avaliações complementares, em todos os lactentes foram detectadas malformações cerebrais compatíveis com a infecção congênita pelo Zika vírus, em 54,5% foi diagnosticado comprometimento ocular e em metade (50,7%) foi observado alguma alteração cardíaca. A principal morbidade clínica apresentada pelas crianças foram as infecções das vias aéreas superiores, seguido de constipação intestinal. Conclusões: Os lactentes com Síndrome da Zika Congênita exibiram comprometimento do crescimento antropométrico, além de grave atraso na aquisição de marcos do desenvolvimento neuromotor. Constatou-se baixa prevalência de aleitamento materno exclusivo até os 6 meses, com alta frequência de dificuldades alimentares. Observou-se também número significativo de lactentes que evoluíram com irritabilidade, convulsão e espasticidade. Os achados deste estudo reforçam a necessidade de acompanhamento multiprofissional especializado para estas crianças, voltado para terapias de reabilitação e apoio aos familiares envolvidos. / Aracaju

Submitted by Geandra Rodrigues (geandrar@gmail.com) on 2018-09-04T12:29:27Z No. of bitstreams: 0 / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2018-09-04T13:24:20Z (GMT) No. of bitstreams: 0 / Made available in DSpace on 2018-09-04T13:24:20Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-07-19 / No ano de 2015, o Brasil enfrentou um surto de infecção por Zika Vírus (ZV), principalmente na região Nordeste, sendo 907 casos em gestantes. Após o nascimento destes lactentes, foi confirmada a associação entre o vírus e diversas malformações congênitas, caracterizando a Síndrome da Infecção Congênita do Zika Vírus (SCZV), que pode levar ao comprometimento das funções motoras, cognitivas, auditivas e visuais. O objetivo do presente estudo foi avaliar o desenvolvimento motor, as oportunidades de estimulação presentes no domicílio de lactentes que foram expostos à Infecção por Zika Vírus na gestação durante os primeiros 20 meses de vida e o perfil epidemiológico desta condição em Juiz de Fora e região. Trata-se de um estudo longitudinal, prospectivo, observacional, aprovado pelo Comitê de Ética em Pesquisa com Seres Humanos da Universidade Federal de Juiz de Fora – UFJF sob parecer n°: 2.001.169. O desenvolvimento motor dos participantes foi avaliado através das Escalas Bayley de Desenvolvimento do Bebê e da Criança Pequena- Terceira Edição (Bayley III) e as oportunidades de estimulação presentes no ambiente domiciliar através do questionário Affordances in the Home Environment for motor Development- Infant Scale (AHEMD-IS) e Self Report (AHEMD-SR). Na descrição do perfil epidemiológico, o presente estudo observou maior taxa de infeção no sexo feminino e na faixa etária de 21 a 41 anos. Quanto ao desenvolvimento motor, cinco participantes apresentaram algum tipo de atraso ou alteração em pelo menos uma das avaliações realizadas, sendo que somente dois foram enquadrados ao nascimento como SCZV. Nenhum participante apresentou microcefalia ao nascimento, porém dois desenvolveram alterações das medidas cefálicas nos meses subsequentes. Foi encontrada correlação forte e moderada entre a Escala Motora da Bayley e o AHEMD nas faixas etárias de 8 a 12 meses e de 17 a 20 meses, respectivamente. Com base nesses resultados conclui-se que a infecção por ZV em Juiz de Fora teve grande potencial de afetar gestantes, pois a maior prevalência foi no sexo feminino e em idade reprodutiva, podendo gerar desfechos desfavoráveis ao feto, mesmo em casos que não foram enquadrados ao nascimento como SCZV, pois foram observados altos porcentuais de atrasos ou alterações no desenvolvimento motor dos participantes do estudo. As relações de moderadas a fortes entre AHEMD e Bayley reforçam a importância do ambiente no desenvolvimento motor nos primeiros anos de vida, devendo ser dada atenção especial às oportunidades de estimulação presentes no domicílio dos casos onde houve infecção gestacional por ZV. / In the year 2015, Brazil faced an outbreak of infection by Zika Virus (ZV), mainly in the Northeast region, with 907 cases in pregnant women. After the birth of these infants, the association between the virus and several congenital malformations was confirmed, characterizing the Congenital Zika Virus Syndrome (SCZV), which can lead to impaired motor, cognitive, auditory and visual functions. The objective of the present study was to evaluate the motor development, the stimulation opportunities present in the domicile of infants who were exposed to Zika Virus infection during gestation during the first 20 months of life and the epidemiological profile of this condition in Juiz de Fora and region. This is a longitudinal, prospective, observational study, approved by the Human Research Ethics Committee of the Federal University of Juiz de Fora - UFJF under opinion no: 2,001,169. Participants' motor development was assessed using the Bayley Scale of Infant and Toddler Development (Bayley III) and the stimulation opportunities present in the home environment through the Affordances in the Home Environment for Motor Development - Infant Scale (AHEMD-IS) and Self Report (AHEMD-SR). In the description of the epidemiological profile, the present study observed a higher rate of infection in the female sex and in the age group of 21 to 41 years. Regarding motor development, five participants presented some type of delay or alteration in at least one of the evaluations performed, and only two were classified at birth as SCZV. No participant presented microcephaly at birth, but two developed changes in cephalic measurements in the subsequent months. A strong and moderate correlation was found between the Bayley Motor Scale and AHEMD in the 8 to 12 months and 17 to 20 months age groups, respectively. Based on these results, it is concluded that ZV infection in Juiz de Fora had a great potential to affect pregnant women, since the highest prevalence was in females and reproductive age, and could result in unfavorable outcomes to the fetus, even in cases that were not framed at birth as SCZV, as high percentages of delays or motor development changes were observed in the study participants. The moderate to strong relationships between AHEMD and Bayley reinforce the importance of the environment in motor development in the first years of life, and special attention should be given to the stimulation opportunities present in the domicile of cases where there was gestational ZV infection.

INTRODUÇÃO: Houve uma epidemia de infecção pelo vírus Zika (ZIKV) no Brasil, entre 2015 e 2016, que refletiu no aumento de casos de microcefalia neonatal secundária à infecção congênita pelo ZIKV. Para compreensão do padrão de acometimento neurológico dessas crianças foi realizado um projeto de pesquisa pelo Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto da Universidade de São Paulo (HCFMRP-USP), conhecido como ZIG, e realizado seguimento neurológico desses pacientes por meio do Exame Neurológico de Hammersmith Neonatal (HINE-N). OBJETIVOS: Avaliar pacientes com microcefalia secundária à infecção congênita pelo ZIKV por meio do HINE- N, descrever as características clínicas e de neuroimagem dos pacientes estudados e identificar fatores pré-natais, perinatais e demográficos potencialmente relacionados ao grau de comprometimento do exame neurológico. METODOLOGIA: O presente estudo faz parte do Núcleo de Estudos sobre Infecção Materna, Perinatal e Infantil (NEIMPI) do HCFMRP-USP e trata-se de um estudo não experimental, transversal, com intuito de correlacionar variáveis qualitativas e quantitativas. Participaram do estudo crianças com microcefalia secundária à infecção congênita pelo ZKV nascidas no município de Ribeirão Preto e seu Departamento Regional de Saúde (DRS XIII), no período de outubro de 2015 a dezembro de 2016. Os dados foram coletados no período de outubro de 2015 a janeiro de 2017. Realizou-se avaliação do perímetro craniano (PC), aplicação do HINE e coleta de dados em prontuário. Os dados foram processados e analisados com a utilização do Pacote Estatístico Statistical Package for the Social Sciences (SPSS), versão 22.0. RESULTADOS: Foram avaliadas 18 crianças, sendo predomínio do sexo feminino (55,6%), nascidas a termo, com exceção de uma que nasceu com 32 semanas, maioria originária de Ribeirão Preto (66,7%), via de parto normal (72,2%), ocorrido no Centro Obstétrico do HCFMRP - USP (83,3%), todas as crianças tiveram APGAR maior que 7. O peso de nascimento foi entre 1228 e 3200 gramas, com PC entre 26 e 31,5 cm. 72,2% das crianças fizeram ultrassom transfontanela e 100% fizeram ressonância magnética de encéfalo, com associação entre os resultados encontrados neles (ambos com p<ou=0,001). O score HINE-N foi baixo, porém não foram encontradas associações entre o score e as variáveis clínicas (ambos com p>0,05). Evidenciou-se associação entre a realização de pré-natal e a cidade de residência, local de nascimento, diagnóstico de microcefalia e de infecção pelo ZIKV no pré-natal, tipo de parto (p< ou = 0,001). A medida do PC apresentou associação direta com a presença de crise convulsiva (p<0,05) e houve associação entre crise convulsiva, ventriculomegalia e calcificação (p< ou= 0,001). CONCLUSÃO: O padrão de acometimento neurológico das crianças com microcefalia secundária à infecção congênita pelo ZIKV é muito severo, com presença de sinais precoces de paralisia cerebral desde o exame neonatal. Este estudo sugere seguimento prospectivo desses pacientes para melhor avaliação prognóstica e associação do score HINE com condições clínicas pré e perinatais / INTRODUCTION: There was an epidemic of Zika virus infection (ZIKV) in Brazil between 2015 and 2016, which reflected the increase in cases of neonatal microcephaly secondary to congenital ZIKV infection. A study was carried out for the Hospital das Clínicas of the University of São Paulo at Ribeirão Preto Medical School (HCFMRP-USP), and the followup of these patients was performed using the Neonatal Hammersmith Neurological Exam (HINE-N) aiming to understand the neurological involvement pattern of these children. OBJECTIVES: To evaluate patients with microcephaly secondary to ZIKV congenital infection using HINE-N, to describe the clinical and neuroimaging characteristics of the patients studied, and to identify prenatal, perinatal and demographic factors potentially related to the degree of impairment of neurological examination. METHODOLOGY: This study is part of the Center for Studies on Maternal, Perinatal and Child Infection (NEIMPI) of the HCFMRP-USP. It is a non-experimental, cross-sectional study aimed at correlating qualitative and quantitative variables. Children with microcephaly secondary to congenital ZKV infection born in the city of Ribeirão Preto and its Regional Health Department (DRS XIII), from October 2015 to December 2016, participated in the study. Data were collected in the period of October 2015 to January 2017. An evaluation of the cranial perimeter (PC), the application of HINE and data collection in medical records were performed. Data were processed and analyzed using the Statistical Package for Social Statistical Package (SPSS), version 22.0. RESULTS: A total of 18 children were evaluated, being 55,6% female, all to term with the exception of one who was born at 32 weeks, most of them from Ribeirão Preto (66.7%), normal delivery (72.2%), occurred in the Obstetric Center of the HCFMRP - USP (83.3%), all children had APGAR greater than 7. The birth weight was between 1228 and 3200 grams, with PC between 26 and 31.5 cm. 72.2% of the children had transfontanel ultrasonography and 100% had brain magnetic resonance imaging, with an association between the results found in them (both with p <or = 0.001). The HINE-N score was low, but no associations were found between the score and the clinical variables (both with p> 0.05). It was evidenced an association between prenatal and city of residence, place of birth, diagnosis of microcephaly and ZIKV infection in the prenatal period, type of delivery (p <or = 0.001). The PC measurement presented a direct association with the presence of seizures (p <0.05) and there was an association between seizure, ventriculomegaly and calcification (p <or = 0.001). CONCLUSION: The pattern of neurological involvement of children with microcephaly secondary to congenital ZIKV infection is very severe, with early signs of cerebral palsy since the neonatal examination. This study suggests a prospective follow-up of these patients for better prognostic evaluation and association of the HINE score with pre and perinatal clinical conditions