As the current growing threat to maternal-fetal health, the most recent and largest outbreak of the Zika virus has introduced the devastating fetal effects of microcephaly and other central nervous system deficits. Therefore, the need for appropriate recognition, treatment, management, and prevention of the Zika virus prompts the necessity for further education and high quality level research to be conducted and utilized. A search of the literature using the databases PubMed, UptoDate, and CINAHL was conducted for articles published between 2009 and 2016. In addition, key informant interviews from various specialties including clinical genetics and public health were conducted. The proposed best practice recommendations for education regarding the Zika virus and appropriate prevention and treatment methods are outlined in an electronic education module that would be delivered to patients and their families prior to visiting their healthcare providers. As the Zika virus continues to spread and further research is conducted regarding its teratogenic effects, the need for concise and effective education is critical in order to raise awareness and conversely decrease the potential for maternal exposure and adverse fetal outcomes.

Objective: To report an outbreak of ZIKV infection among asymptomatic pregnant women during 2016 in the city of Jaen, Cajamarca. Results: Zika virus RNA was detected in 3.2% (n = 36) of cases by RT-PCR. The mean age of patients positive for ZIKV infection was 29.6 years. 7 patients (19.4%) infected with ZIKV were in their first-trimester of gestation, 13 (36.1%) were in their second-trimester, and 16 (44%) were in their third-trimester. All of the infected pregnant women were asymptomatic. ZIKV infection remains a major public health issue that calls for constant epidemiological surveillance. It can cause the congenital Zika virus syndrome in the newborns of infected mothers. The lack of molecular diagnostic methods in isolated localities and the similarity of symptoms to other arboviral infections, lead to an under-diagnosis of this disease in endemic areas. / Revisión por pares

Orientador: Rejane Maria Tommasini Grotto / Resumo: O vírus Zika pertence a família Flaviviridae, gênero Flavivirus e tem seu genoma viral constituído de uma molécula de RNA de fita simples de polaridade positiva, com aproximadamente 10.700 pb. As interações de Flavivirus com compartimentos biológicos como as plaquetas foram relatadas pela literatura, sendo demostrado que o vírus é carreado para outros tecidos e sua viabilidade é mantida por um maior tempo, no entanto, este tipo de estudo ainda não foi reportado para o ZIKV. O objetivo deste trabalho foi comparar a carga viral do vírus Zika entre o plasma rico em plaquetas (PRP) e o plasma pobre em plaquetas (PPP). As amostras utilizadas foram coletadas de doadores saudáveis e posteriormente incubadas com o vírus ZIKA à 37°C na presença de CO2 durante 12 dias consecutivos, sendo retiradas alíquotas no momento do preparo e após 2, 4, 6, 8, 10 e 12 dias de incubação. Os resultados demostraram que a carga viral no PRP foi estatisticamente superior a do PPP depois de 12 dias de incubação com o vírus, sugerindo que as plaquetas podem carrear e preservar o vírus durante maior tempo comparado ao plasma isento de plaquetas. / Abstract: The Zika virus belongs to the Flaviviridae family, Flavivirus genus, and the viral genome consists of the molecule single-stranded, positive-sense RNA of approximately 10,700 bp. The interaction of Flavivirus with biological compartments such as platelets has been reported in the literature, and have shown that the virus is carried to other tissues and its viability is maintained for a longer time. However, this kind of evaluation was not reported for ZIKV yet. The aim of this study was to compare the viral load of Zika virus between platelet rich plasma (PRP) and platelet poor plasma (PPP). The samples were collected from healthy donors and subsequently incubated with ZIKA virus at 37 ° C with CO2 atmosphere for 12 consecutive days. Aliquots were evaluated at the zero time and after 2, 4, 6, 8, 10 and 12 days of incubation Results showed that the viral load in PRP is statistically higher than in PPP after 12 days of incubation with the virus, suggesting that platelets can carry and preserve the virus longer than in platelet-free plasma. / Mestre

archives@tulane.edu / Brazil was at the epicenter of the 2015-2016 Zika virus (ZIKV) epidemic, with approximately 60% of globally reported cases within its borders. This dissertation explored how contextual factors were relevant to women’s ability and desire to adhere to the ZIKV recommendations made by the Brazilian Ministry of Health during the epidemic. More specifically this dissertation focused on women’s response to the recommendation to avoid pregnancy during this epidemic. Methods utilized in this dissertation included a rapid assessment and a series of case studies of pregnancy narratives from women in Fortaleza, Brazil. By more deeply exploring women’s fertility decision making during the ZIKV epidemic, a more comprehensive understanding of the underlying contextual factors influencing women’s fertility desires, decisions, and behaviors was developed. Among the findings, we document that the health recommendations promoted during the epidemic were made without the use of best practices in health communication and without acknowledgement of the context in which the epidemic was taking place. For example, these recommendations assumed women had the power to adhere to health authorities’ directives to prevent ZIKV. Furthermore, we document how these generic recommendations ignored the unique conditions of vulnerable populations constrained by income, class, and gender. Analysis showed that participants perceived the recommendations as unrealistic, recycled, unhelpful, or not directed toward them. Case studies and pregnancy narratives illuminated major gaps in the healthcare and support provided to pregnant women during the outbreak, as well as to the mothers of children born with Congenital Zika Syndrome (CZS). ZIKV is the latest of an inevitable influx of emerging infectious diseases that are being brought on by deforestation, urbanization, and global warming. With these diseases on the horizon, it is important to acknowledge and research the underlying contextual factors that permeate these epidemics and continue to devastate the most marginalized within societies. / 1 / Jeni Stolow

archives@tulane.edu / Zika virus (ZIKV), a member of the Flaviviridae family, was the cause of a large viral outbreak reaching across the Americas during 2015 and 2016. Discovered in 1947, it has historically been a neglected disease, due to its emergence in humans on a large scale being recent. At the time of the outbreak, no FDA approved ZIKV diagnostics existed, and those that were able to detect the virus were unable to distinguish it from related viruses such as Dengue virus (DENV), and at this time, no approved therapeutics or vaccines are available. We investigated the ability of diagnostics targeted toward both anti-NS1 antibodies and NS1 antigen circulating during infection to detect current or past ZIKV disease, as well as the capability of NS1 to produce a protective response. Our studies suggest anti-NS1diagnostics are feasible, though some populations may display an immune response reminiscent of a prior infection. Levels of circulating NS1 were lower than those produced during DENV infection, though were still detectable with our assay. Additionally, intraperitoneal immunization with NS1 produced an anti-ZIKV NS1 response that coincided with a decrease in viremia, though further work was needed to discern life-prolonging effects. Together, this work furthers the development of the tools necessary to combat future outbreaks of ZIKV in vulnerable populations. / 1 / Brandon Beddingfield

abstract: Zika virus (ZIKV) outbreaks have been linked to several neurological pathologies in the developing fetus, which can progress to spontaneous abortion and microcephaly in newborns whose mothers were infected with the virus during pregnancy. ZIKV has also been correlated with neurological complications in adults such as Guillain-Barré Syndrome (GBS). ZIKV outbreaks often occur in low income areas with limited access to healthcare. Therefore, there is a need to create a low-cost preventative vaccine against the virus. Mature ZIKV particles contain a lipid bilayer, a positive sense single stranded RNA genome and three structural proteins: the envelope (E), membrane (M) and capsid (C) proteins. Congruently, to other members of the Flaviviridae family, ZIKV proteins are synthesized as a polyprotein precursor which needs to be processed to release the mature structural and non-structural viral proteins. Past studies have determined the ZIKV precursor protein is cleaved by a host furin protease which separates the Pr peptide and the M protein, while the host signal peptidase separates the M and E protein. Processing is important for correct folding of the E protein. In turn, the most important neutralizing antibodies upon infection are directed against epitopes of the E protein. In this work, we used a Bean Yellow Dwarf Viral vector system to transiently express, in Nicotiana benthamiana plants, a portion of the ZIKV polyprotein encoding the Pr, M and E proteins. I further demonstrate that plants can proteolytically process the polyprotein to yield the two integral membrane proteins M and E. These proteins can be shown to co-partition into a soluble membrane-particulate fraction, consistent with formation of enveloped virus-like particles (VLPs). This work provides the first step in creating a low-cost sustainable plant-based production system of ZIKV VLPs that can be explored as a potential component 0f a low-cost prophylactic vaccine against ZIKV. / Dissertation/Thesis / Masters Thesis Molecular and Cellular Biology 2018

No Brasil os vírus Zika e Chikungunya vem causando enormes epidemias com grande morbidade e significante mortalidade, tornou-se importante desenvolver métodos diagnósticos eficazes para discrimar suas infecções. No presente trabalho, desenvolvemos ensaios imunoenzimáticos utilizando células de cultura, aedes albopictus pseudo C6/36, infectadas e utilizadas como antígeno viral (EIA-ICC), para diagnósticos de infecções provocadas pelos vírus Zika (ZIKV) e Chikungunya (CHIKV). As placas contendo células infectadas tiveram suas antigenicidades verificadas nos ensaios utilizando anticorpos policlonais oriundo de camundongos hiperimunizados com ZIKV, CHIKV e outros flavivirus e alphavirus. O EIAICC ZIKV detectou anticorpos murinos de forma homotípica, porém reações cruzadas foram encontradas com Dengue 1 (DENV 1), Saint Louis Vírus (SLEV), Rocio Vírus (ROCV) e West Nilo Vírus (WNV). O EIA-ICC CHIKV também detectou anticorpos murinos de forma homotípica e reações cruzadas foram observadas com os Mayaro Vírus (MAYV) e Mucambo Vírus (MUCV). Resultados comparativos de 39 amostras foram realizados no EIA-ICC ZIKV, que demonstrou sensibilidade de 87,5% (IgM) e 100% (IgG) e especificidade de 95,65% (IgM) e 83,33% (IgG). O EIA-ICC CHIKV, teste comparativos entre 59 amostras, demostrou sensibilidade de 64,7% (IgM) e 92,3% (IgG) e especificidade de 40% (IgM) e 96,96% (IgG). Os EIA-ICCs também foram usados para diagnósticos das arboviroses ZIKV e CHIKV em 174 soros de pacientes com doença febril aguda, no EIA-ICC ZIKV foi encontrado 8,13% de IgM, 20,34% IgG e 6,97% de IgG e IgM na mesmo soro, no EIA-ICC CHIKV encontramos IgG em 8,13% dos soros e IgG em 16,86%, soros com anticorpos IgG e IgM nos mesmos foram vistos em 1,74%. Também encontramos em 11 soros anticorpos tanto para ZIKV quanto para CHIKV. Portanto o EIA-ICC ZIKV mostrou para detecção de IgM e IgG, adequadamente sensível e específico para ser validado em estudos com maiores números de amostras e ser aplicados no diagnóstico. O EIA-ICC CHIKV mostrou para detecção de IgG, adequadamente sensível e específico para ser validado em estudos com maiores números de amostras e ser aplicados no diagnóstico. Porém não é indicado para a detecção de IgM / In Brazil, Zika and Chikungunya viruses have been causing huge epidemics with great morbidity and significant mortality. It\'s important to develop effective diagnostic methods to discriminate against their infections. In the present study, we developed immunoenzymatic assays using cultured cells from aedes albopictus C6 / 36 infected and used as viral antigen (EIA-ICC), for diagnosis of infections caused by the Zika (ZIKV) and Chikungunya (CHIKV) viruses. Plates containing infected cells had their antigenicities verified in the assays using polyclonal antibodies from mice hyperimmunized with ZIKV, CHIKV and other flaviviruses and alphaviruses. The EIA-ICC ZIKV detected murine antibodies in a homotypic manner, but cross-reactions were found with Dengue 1 virus (DENV 1), Saint Louis virus (SLEV), Rocio virus (ROCV) and West Nile virus (WNV). The EIA-ICC CHIKV also detected murine antibodies in a homotypic manner and cross-reactions were observed with the Mayaro virus (MAYV) and Mucambo virus (MUCV). Comparative results of 39 samples were performed in the EIA-ICC ZIKV, which showed sensitivity of 87.5% (IgM) and 100% (IgG) and specificity of 95.65% (IgM) and 83.33% (IgG). The EIA-ICC CHIKV, a comparative test among 59 samples, showed a sensitivity of 64.7% (IgM) and 92.3% (IgG) and a specificity of 40% (IgM) and 96.96% (IgG). The EIA-ICCs were also used for diagnosis of ZIKV and CHIKV arboviruses in 174 sera from patients with acute febrile illness. In the EIA-ICC, ZIKV found 8.13% IgM, 20.34% IgG and 6.97% IgG and IgM in the same serum, in the EIA-ICC CHIKV we found IgG in 8.13% of the sera and IgG in 16.86%, sera with IgG and IgM antibodies in them were seen in 1.74%. We also found 11 samples positive for both ZIKV and CHIKV. Therefore, the EIA-ICC ZIKV showed for detection of IgM and IgG, adequately sensitive and specific to be validated in studies with larger numbers of samples and to be applied in the diagnosis. The EIA-ICC CHIKV showed for IgG detection, adequately sensitive and specific to be validated in studies with larger numbers of samples and to be applied in the diagnosis. However, it is not indicated for the detection of IgM

O Vírus Zika é um arbovírus pertencente à família Flaviviridae e ao gênero Flavivirus, é constituído por RNA de cadeia simples senso positivo e foi primeiramente descrito em 1947 na floresta de Ziika, em Uganda. Este estudo, visa mapear os epitopos do Vírus Zika (ZIKV), experimento que nos permite avaliar e compreender a resposta imune do hospedeiro contra o vírus por meio da detecção de regiões imunodominantes nas proteínas virais. O processo de identificação dos epitopos de ZIKV foi realizado pela técnica de spot synthesis, que se baseia em arranjos de peptídeos relacionados às proteínas do ZIKV. Amostras de soro de pacientes residentes em áreas endêmicas de arboviroses, mães e bebês que apresentaram sintomatologia típica foram selecionadas e triadas pelo método de ELISA e PCR real time. Sequências de referência do vírus foram submetidas a análises in sílico, para a determinação das áreas de interação na superfície da proteína e suas características antigênicas, para efeito de comparação com a resposta observada pelas amostras. Os experimentos evidenciaram um alta antigenicidade nas proteínas estruturais do vírus, e em nas não-estruturais 1,3 e 5. / The Zika virus is an arbovirus belonging to the genus Flavivirus of the Flaviridae family. It consists of single-stranded positive-sense RNA and was first described in 1947 in the Ziika Forest in Uganda. This study aims to map the Zika virus (ZIKV) epitopes, in an experimental design that allows us to evaluate and understand the immune response of the host against the virus by detecting immunodominant regions in the viral proteins. The process of identification of the ZIKV epitopes was performed by the spot synthesis technique, which is based on peptide arrangements related to the ZIKV proteins. Serum samples from patients living in arboviruses endemic areas, mothers and infants presenting typical symptoms were selected and screened by ELISA and real time PCR. Reference sequences of the virus were submitted to in silico analyzes to determine the interaction areas on the surface of the protein and its antigenic characteristics, for comparison with the response observed by using serum samples. The experiments demonstrated a high antigenicity in the structural proteins of the virus, and in the non-structural proteins 1, 3 and 5.

<p>Our ability to convert basic knowledge into robust anti-viral therapeutics requires discovery of novel host-virus interactions as well as an informed anti-viral discovery pipeline. We used a genome-scale RNAi-based screen followed by a chemical screen of FDA-approved therapeutics to identify scores of novel dengue virus (DENV) human host dependency factors (HDF) and identified more than 20 potential anti-Zika virus (ZIKV) therapeutics. </p><p>Two genes in particular, TTC35 and TMEM111, strongly inhibited DENV infection and, based on comparisons with published literature, implicated a larger protein, the ER Membrane Protein Complex (EMC), as a pan-flavivirus HDF. The EMC is a poorly characterized multiprotein complex that may function in ER-associated protein biogenesis and/or lipid metabolism. Based on our screen data, we hypothesized that the EMC is an uncharacterized HDF that functions through a common mechanism to promote replication of flaviviruses. We report that DENV, ZIKV, and yellow fever virus (YFV) infections were impressively inhibited, while West Nile Virus (WNV) infection was unchanged, in cell lines engineered to lack EMC subunit 4 (EMC4). Furthermore, targeted depletion of EMC subunits in live mosquitos significantly reduced DENV-2 propagation in vivo. In addition, the accumulation of DENV proteins shortly after infection in EMC4 knockout cells was significantly reduced, suggesting that the EMC promotes viral protein biogenesis. </p><p>We interrogated a library of FDA-approved drugs for their ability to block infection of human HuH-7 cells by a newly isolated ZIKV strain. Selected compounds were further validated for inhibition of ZIKV infection in human cervical, placental, and neural stem cell lines, as well as primary human amnion cells. Established anti-flaviviral drugs (e.g., bortezomib and mycophenolic acid) and others that had no previously known antiviral activity (e.g., daptomycin) were identified as inhibitors of ZIKV infection. Several drugs reduced ZIKV infection across multiple cell types.</p><p>We propose that the EMC may be exploited as a novel therapeutic target for multiple flaviviruses in the future. Also we identified drugs that could be tested in clinical studies of ZIKV infection and provides a resource of small molecules to study ZIKV pathogenesis.</p> / Dissertation

The presence of El Niño Southern Oscillation (ENSO) implies the presence of fluctuating rains in coastal areas and these changes influence the occurrence of febrile syndromes outbreaks. In Peru, Aedes aegypti is the vector responsible for various viruses such as the dengue, Zika, chikungunya, which is distributed in 18 Peruvian departments. These viruses cause similar clinical characteristics in the host and for this reason rapid, sensitive and specific diagnostic tests are needed so that the patient can receive timely treatment. / Revisión por pares