Association between Maternal Non-Coding Interferon-λ Polymorphisms and Congenital Zika Syndrome in a Cohort from Brazilian Northeast.

Congenital Zika syndrome (CZS) is characterized by a diverse group of congenital malformations induced by ZIKV infection during pregnancy. Type III interferons have been associated with placental immunity against ZIKV and restriction of vertical transmission in mice, and non-coding single-nucleotide polymorphisms (SNPs) on these genes are well known to influence susceptibility to other viral infections. However, their effect on ZIKV pathogenesis has not yet been explored. To investigate whether maternal non-coding SNPs at IFNL genes are associated with CZS, 52 women infected with ZIKV during pregnancy were enrolled in a case-control association study. A total of 28 women were classified as cases and 24 as controls based on the presence or absence of CZS in their infants, and seven Interferon-λ non-coding SNPs (rs12980275, rs8099917, rs4803217, rs4803219, rs8119886, rs368234815, rs12979860) were genotyped. The results of logistic regression analyses show an association between the G allele at rs8099917 and increased susceptibility to CZS under a log-additive model (adjustedOR = 2.80; 95%CI = 1.14-6.91; p = 0.02), after adjustment for trimester of infection and genetic ancestry. These results provide evidence of an association between Interferon-λ SNPs and CZS, suggesting rs8099917 as a promising candidate for further studies on larger cohorts.

Whole-exome sequencing reveals insights into genetic susceptibility to Congenital Zika Syndrome.

Congenital Zika Syndrome (CZS) is a critical illness with a wide range of severity caused by Zika virus (ZIKV) infection during pregnancy. Life-threatening neurodevelopmental dysfunctions are among the most common phenotypes observed in affected newborns. Risk factors that contribute to susceptibility and response to ZIKV infection may be related to the virus itself, the environment, and maternal genetic background. Nevertheless, the newborn's genetic contribution to the critical illness is still not elucidated. Here, we aimed to identify possible genetic variants as well as relevant biological pathways that might be associated with CZS phenotypes. For this purpose, we performed a whole-exome sequencing in 40 children born to women with confirmed exposure to ZIKV during pregnancy. We investigated the occurrence of rare harmful single-nucleotide variants (SNVs) possibly associated with inborn errors in genes ontologically related to CZS phenotypes. Moreover, an exome-wide association analysis was also performed using a case-control design (29 CZS cases and 11 controls), for both common and rare variants. Five out of the 29 CZS patients harbored known pathogenic variants likely to contribute to mild to severe manifestations observed. Approximately, 30% of affected individuals carried at least one pathogenic or likely pathogenic SNV in genes candidates to play a role in CZS. Our common variant association analysis detected a suggestive protective effect of the rs2076469 in DISP3 gene (p-value: 1.39 x 10-5). The IL12RB2 gene (p-value: 2.18x10-11) also showed an unusual distribution of nonsynonymous rare SNVs in control samples. Finally, genes harboring harmful variants are involved in processes related to CZS phenotypes such as neurological development and immunity. Therefore, both rare and common variations may be likely to contribute as the underlying genetic cause of CZS susceptibility. The variations and pathways identified in this study may also have implications for the development of therapeutic strategies in the future.

Brazilian Protocol for Sexually Transmitted Infections 2020: viral hepatitis.

This article discusses viral hepatitis, a theme addressed by the Clinical Protocol and Therapeutic Guidelines to Comprehensive Care for People with Sexually Transmitted Infections and, more precisely, by the Clinical Protocols and Therapeutic Guidelines for Hepatitis B and Hepatitis C and Coinfections, published by the Brazilian Ministry of Health. Besides the broad spectrum of health impairment, hepatitis A, B, and C viruses also present different transmission forms, whether parenteral, sexual, vertical, or fecal-oral. Among the strategies suggested for the control of viral hepatitis, in addition to behavioral measures, are expanded diagnosis, early vaccination against hepatitis A and hepatitis B viruses, and access to available therapeutic resources. Considering vertical transmission of the hepatitis B and hepatitis C viruses, screening for pregnant women with chronic hepatitis B and C is an essential perinatal health strategy, indicating with precision those who can benefit from the prophylactic interventions. Viral hepatitis A, B, and C are responsible for more than 1.34 million deaths worldwide every year, from which 66% are the result of hepatitis B, 30% of hepatitis C, and 4% of hepatitis A.

[Brazilian Protocol for Sexually Transmitted Infections 2020: viral hepatitis]. / Protocolo Brasileiro para Infecções Sexualmente Transmissíveis 2020: hepatites virais.

This article discusses viral hepatitis, a theme addressed by the Clinical Protocol and Therapeutic Guidelines to Comprehensive Care for People with Sexually Transmitted Infections and, more precisely, by the Clinical Protocols and Therapeutic Guidelines for Hepatitis B and Hepatitis C and Coinfections, published by the Brazilian Ministry of Health. Besides the broad spectrum of health impairment, hepatitis A, B and C viruses also present different forms of transmission, whether parenteral, sexual, vertical or oral. Among the strategies suggested for the control of viral hepatitis, in addition to behavioral measures, are expanded diagnosis, early vaccination against hepatitis A and hepatitis B viruses, and access to available therapeutic resources. Considering vertical transmission of the hepatitis B and hepatitis C viruses, screening for pregnant women with chronic hepatitis B and C is an important perinatal health strategy, indicating with precision those who can benefit from the prophylactic interventions.

Este artigo aborda as hepatites virais, tema tratado no Protocolo Clínico e Diretrizes Terapêuticas para Atenção Integral às Pessoas com Infecções Sexualmente Transmissíveis e, mais precisamente, nos Protocolos Clínicos e Diretrizes Terapêuticas para Hepatite B e para Hepatite C e Coinfecções, publicados pelo Ministério da Saúde do Brasil. Além do espectro ampliado de acometimento da saúde, os vírus das hepatites A, B e C também apresentam diferentes formas de transmissão, seja parenteral, sexual, vertical ou oral. Entre as estratégias sugeridas para o controle das hepatites virais, além das medidas comportamentais, estão o diagnóstico ampliado, a vacinação precoce contra os vírus da hepatite A e hepatite B e o acesso aos recursos terapêuticos disponíveis. Considerando a transmissão vertical dos vírus da hepatite B e hepatite C, a triagem das gestantes portadoras crônicas desses vírus é uma importante estratégia de saúde perinatal, indicando com precisão quem pode se beneficiar das intervenções profiláticas disponíveis.

Este artículo aborda las hepatitis virales, tema que hace parte del Protocolo Clínico y Directrices Terapéuticas para la Atención Integral a Personas con Infecciones de Transmisión Sexual y más precisamente de los Protocolos Clínicos y Guías Terapéuticas para Hepatitis B, Hepatitis C y Coinfecciones, publicados por el Ministerio de Salud. Además del amplio espectro de deterioro de la salud, los virus de las hepatitis A, B y C presentan diferentes formas de transmisión, como parenteral, sexual, vertical u oral. Entre las estrategias sugeridas para el control de las hepatitis virales, están las medidas conductuales, el diagnóstico ampliado, la vacunación precoz contra los virus de las hepatitis A y B y el acceso facilitado a los recursos terapéuticos disponibles. Considerando la transmisión vertical de los virus de la hepatitis B y C, la identificación de embarazadas portadoras crónicas de estos virus es importante estrategia de salud perinatal, indicando quiénes pueden beneficiarse de las intervenciones profilácticas.

Protocolo Brasileiro para Infecções Sexualmente Transmissíveis 2020: hepatites virais / Brazilian Protocol for Sexually Transmitted Infections 2020: viral hepatitis / Protocolo Brasileño para las Infecciones de Transmisión Sexual 2020: hepatitis virales

Este artigo aborda as hepatites virais, tema tratado no Protocolo Clínico e Diretrizes Terapêuticas para Atenção Integral às Pessoas com Infecções Sexualmente Transmissíveis e, mais precisamente, nos Protocolos Clínicos e Diretrizes Terapêuticas para Hepatite B e para Hepatite C e Coinfecções, publicados pelo Ministério da Saúde do Brasil. Além do espectro ampliado de acometimento da saúde, os vírus das hepatites A, B e C também apresentam diferentes formas de transmissão, seja parenteral, sexual, vertical ou oral. Entre as estratégias sugeridas para o controle das hepatites virais, além das medidas comportamentais, estão o diagnóstico ampliado, a vacinação precoce contra os vírus da hepatite A e hepatite B e o acesso aos recursos terapêuticos disponíveis. Considerando a transmissão vertical dos vírus da hepatite B e hepatite C, a triagem das gestantes portadoras crônicas desses vírus é uma importante estratégia de saúde perinatal, indicando com precisão quem pode se beneficiar das intervenções profiláticas disponíveis.

This article discusses viral hepatitis, a theme addressed by the Clinical Protocol and Therapeutic Guidelines to Comprehensive Care for People with Sexually Transmitted Infections and, more precisely, by the Clinical Protocols and Therapeutic Guidelines for Hepatitis B and Hepatitis C and Coinfections, published by the Brazilian Ministry of Health. Besides the broad spectrum of health impairment, hepatitis A, B and C viruses also present different forms of transmission, whether parenteral, sexual, vertical or oral. Among the strategies suggested for the control of viral hepatitis, in addition to behavioral measures, are expanded diagnosis, early vaccination against hepatitis A and hepatitis B viruses, and access to available therapeutic resources. Considering vertical transmission of the hepatitis B and hepatitis C viruses, screening for pregnant women with chronic hepatitis B and C is an important perinatal health strategy, indicating with precision those who can benefit from the prophylactic interventions.

Este artículo aborda las hepatitis virales, tema que hace parte del Protocolo Clínico y Directrices Terapéuticas para la Atención Integral a Personas con Infecciones de Transmisión Sexual y más precisamente de los Protocolos Clínicos y Guías Terapéuticas para Hepatitis B, Hepatitis C y Coinfecciones, publicados por el Ministerio de Salud. Además del amplio espectro de deterioro de la salud, los virus de las hepatitis A, B y C presentan diferentes formas de transmisión, como parenteral, sexual, vertical u oral. Entre las estrategias sugeridas para el control de las hepatitis virales, están las medidas conductuales, el diagnóstico ampliado, la vacunación precoz contra los virus de las hepatitis A y B y el acceso facilitado a los recursos terapéuticos disponibles. Considerando la transmisión vertical de los virus de la hepatitis B y C, la identificación de embarazadas portadoras crónicas de estos virus es importante estrategia de salud perinatal, indicando quiénes pueden beneficiarse de las intervenciones profilácticas.

Protocolo Brasileiro para Infecções Sexualmente Transmissíveis 2020: hepatites virais / Brazilian Protocol for Sexually Transmitted Infections 2020: viral hepatitis / Protocolo Brasileño para las Infecciones de Transmisión Sexual 2020: hepatitis virales

Resumo Este artigo aborda as hepatites virais, tema tratado no Protocolo Clínico e Diretrizes Terapêuticas para Atenção Integral às Pessoas com Infecções Sexualmente Transmissíveis e, mais precisamente, nos Protocolos Clínicos e Diretrizes Terapêuticas para Hepatite B e para Hepatite C e Coinfecções, publicados pelo Ministério da Saúde do Brasil. Além do espectro ampliado de acometimento da saúde, os vírus das hepatites A, B e C também apresentam diferentes formas de transmissão, seja parenteral, sexual, vertical ou oral. Entre as estratégias sugeridas para o controle das hepatites virais, além das medidas comportamentais, estão o diagnóstico ampliado, a vacinação precoce contra os vírus da hepatite A e hepatite B e o acesso aos recursos terapêuticos disponíveis. Considerando a transmissão vertical dos vírus da hepatite B e hepatite C, a triagem das gestantes portadoras crônicas desses vírus é uma importante estratégia de saúde perinatal, indicando com precisão quem pode se beneficiar das intervenções profiláticas disponíveis.

Abstract This article discusses viral hepatitis, a theme addressed by the Clinical Protocol and Therapeutic Guidelines to Comprehensive Care for People with Sexually Transmitted Infections and, more precisely, by the Clinical Protocols and Therapeutic Guidelines for Hepatitis B and Hepatitis C and Coinfections, published by the Brazilian Ministry of Health. Besides the broad spectrum of health impairment, hepatitis A, B and C viruses also present different forms of transmission, whether parenteral, sexual, vertical or oral. Among the strategies suggested for the control of viral hepatitis, in addition to behavioral measures, are expanded diagnosis, early vaccination against hepatitis A and hepatitis B viruses, and access to available therapeutic resources. Considering vertical transmission of the hepatitis B and hepatitis C viruses, screening for pregnant women with chronic hepatitis B and C is an important perinatal health strategy, indicating with precision those who can benefit from the prophylactic interventions.

Resumen Este artículo aborda las hepatitis virales, tema que hace parte del Protocolo Clínico y Directrices Terapéuticas para la Atención Integral a Personas con Infecciones de Transmisión Sexual y más precisamente de los Protocolos Clínicos y Guías Terapéuticas para Hepatitis B, Hepatitis C y Coinfecciones, publicados por el Ministerio de Salud. Además del amplio espectro de deterioro de la salud, los virus de las hepatitis A, B y C presentan diferentes formas de transmisión, como parenteral, sexual, vertical u oral. Entre las estrategias sugeridas para el control de las hepatitis virales, están las medidas conductuales, el diagnóstico ampliado, la vacunación precoz contra los virus de las hepatitis A y B y el acceso facilitado a los recursos terapéuticos disponibles. Considerando la transmisión vertical de los virus de la hepatitis B y C, la identificación de embarazadas portadoras crónicas de estos virus es importante estrategia de salud perinatal, indicando quiénes pueden beneficiarse de las intervenciones profilácticas.

Brazilian protocol for sexually transmitted infections 2020: viral hepatitis

Abstract This article discusses viral hepatitis, a theme addressed by the Clinical Protocol and Therapeutic Guidelines to Comprehensive Care for People with Sexually Transmitted Infections and, more precisely, by the Clinical Protocols and Therapeutic Guidelines for Hepatitis B and Hepatitis C and Coinfections, published by the Brazilian Ministry of Health. Besides the broad spectrum of health impairment, hepatitis A, B, and C viruses also present different transmission forms, whether parenteral, sexual, vertical, or fecal-oral. Among the strategies suggested for the control of viral hepatitis, in addition to behavioral measures, are expanded diagnosis, early vaccination against hepatitis A and hepatitis B viruses, and access to available therapeutic resources. Considering vertical transmission of the hepatitis B and hepatitis C viruses, screening for pregnant women with chronic hepatitis B and C is an essential perinatal health strategy, indicating with precision those who can benefit from the prophylactic interventions.

Laboratory Acquired Zika Virus Infection Through Mouse Bite: A Case Report.

Zika virus is an arthropod-borne flavivirus mainly transmitted by the bite of infected mosquitoes. However, alternative transmission routes can occur. In this study, we show the accidental transmission of virus from an infected mouse to a human during the experimental manipulation. This study describes the patient clinical manifestations and virus genome identification.

Molecular alterations in the extracellular matrix in the brains of newborns with congenital Zika syndrome.

Zika virus (ZIKV) infection during pregnancy can cause a set of severe abnormalities in the fetus known as congenital Zika syndrome (CZS). Experiments with animal models and in vitro systems have substantially contributed to our understanding of the pathophysiology of ZIKV infection. Here, to investigate the molecular basis of CZS in humans, we used a systems biology approach to integrate transcriptomic, proteomic, and genomic data from the postmortem brains of neonates with CZS. We observed that collagens were greatly reduced in expression in CZS brains at both the RNA and protein levels and that neonates with CZS had several single-nucleotide polymorphisms in collagen-encoding genes that are associated with osteogenesis imperfecta and arthrogryposis. These findings were validated by immunohistochemistry and comparative analysis of collagen abundance in ZIKV-infected and uninfected samples. In addition, we showed a ZIKV-dependent increase in the expression of cell adhesion factors that are essential for neurite outgrowth and axon guidance, findings that are consistent with the neuronal migration defects observed in CZS. Together, these findings provide insights into the underlying molecular alterations in the ZIKV-infected brain and reveal host genes associated with CZS susceptibility.

Molecular alterations in the extracellular matrix in the brains of newborns with congenital Zika syndrome

Zika virus (ZIKV) infection during pregnancy can cause a set of severe abnormalities in the fetus known as congenital Zika syndrome (CZS). Experiments with animal models and in vitro systems have substantially contributed to our understanding of the pathophysiology of ZIKV infection. Here, to investigate the molecular basis of CZS in humans, we used a systems biology approach to integrate transcriptomic, proteomic, and genomic data from the postmortem brains of neonates with CZS. We observed that collagens were greatly reduced in expression in CZS brains at both the RNA and protein levels and that neonates with CZS had several single-nucleotide polymorphisms in collagen-encoding genes that are associated with osteogenesis imperfecta and arthrogryposis. These findings were validated by immunohistochemistry and comparative analysis of collagen abundance in ZIKV-infected and uninfected samples. In addition, we showed a ZIKV-dependent increase in the expression of cell adhesion factors that are essential for neurite outgrowth and axon guidance, findings that are consistent with the neuronal migration defects observed in CZS. Together, these findings provide insights into the underlying molecular alterations in the ZIKV-infected brain and reveal host genes associated with CZS susceptibility.

Molecular alterations in the extracellular matrix in the brains of newborns with congenital Zika syndrome

Zika virus (ZIKV) infection during pregnancy can cause a set of severe abnormalities in the fetus known as congenital Zika syndrome (CZS). Experiments with animal models and in vitro systems have substantially contributed to our understanding of the pathophysiology of ZIKV infection. Here, to investigate the molecular basis of CZS in humans, we used a systems biology approach to integrate transcriptomic, proteomic, and genomic data from the postmortem brains of neonates with CZS. We observed that collagens were greatly reduced in expression in CZS brains at both the RNA and protein levels and that neonates with CZS had several single-nucleotide polymorphisms in collagen-encoding genes that are associated with osteogenesis imperfecta and arthrogryposis. These findings were validated by immunohistochemistry and comparative analysis of collagen abundance in ZIKV-infected and uninfected samples. In addition, we showed a ZIKV-dependent increase in the expression of cell adhesion factors that are essential for neurite outgrowth and axon guidance, findings that are consistent with the neuronal migration defects observed in CZS. Together, these findings provide insights into the underlying molecular alterations in the ZIKV-infected brain and reveal host genes associated with CZS susceptibility.

Detection of sexually transmitted infections at a Brazilian gynecology center: high prevalence of co-infections / Detecção de infecções sexualmente transmissíveis em uma unidade ginecológica brasileira: alta prevalência de coinfecções

ABSTRACT Introduction: Sexually transmitted infections (STI) remain a major public health problem and surveillance is crucial for prevention and control strategies. Objective: Our aim was to assess the prevalence of STI in a reference center for gynecology in Rio de Janeiro, Brazil. Materials and methods: It is a cross-sectional study conducted between August 2016 and June 2017. Whole blood and cervical cells were collected from 62 women and tested for human papillomavirus (HPV), human immunodeficiency virus (HIV), syphilis, chlamydia, gonorrhea and herpes simples virus 1 and 2 (HSV-1/2). Cervical lesions were diagnosed by cytopathology and in some patients by colposcopy (79%). Other STI were evaluated during clinical examination. Results: Cervical lesions were detected by cytopathology examinations in 46.8% of patients; those with a history of four sexual partners were at higher risk of developing them. There was moderate agreement between the cytopathology e colposcopy results (Kappa = 0.69). The prevalence of HSV (96.7%), syphilis (6.4%) and HIV (3.2%) were higher than that described in the literature, while the prevalence of chlamydia (6.4%) and gonorrhea (1.6%) were similar. HPV was detected in 53.2% of women, 32.3% of which were infected by HPV 16. In the context of co-infections, 38 women (61.3%) presented more than one STI. Conclusion: Since most of the women analyzed were affected by more than one STI, our results suggest that routine screening for these infections at health centers would help in early detection, treatment and prevention of these infections. These measures would also impact on patients' cervical cancer control.

RESUMO Introdução: As infecções sexualmente transmissíveis (ISTs) ainda são um importante problema de saúde pública, e a vigilância é essencial para sua prevenção e seu controle. Objetivo: Avaliar a prevalência das ISTs em uma unidade de referência ginecológica no Rio de Janeiro, Brasil. Materiais e métodos: Trata-se de um estudo transversal realizado entre agosto de 2016 e junho de 2017. Foram coletados sangue total e células cervicais de 62 mulheres e realizados testes para detecção de papiloma vírus humano (HPV), vírus da imunodeficiência humana (HIV), sífilis, clamídia, gonorreia e vírus do herpes simples 1 e 2 (HSV-1/2). As lesões cervicais foram diagnosticadas por citopatologia, e em algumas pacientes, por colposcopia (79%). Outras ISTs foram avaliadas durante o exame clínico. Resultados: Foram encontradas lesões cervicais pelo exame citopatológico em 46,8% das pacientes; aquelas com história de quatro parceiros sexuais tiveram mais risco de desenvolvê-las. Houve concordância moderada entre os resultados da citopatologia e colposcopia (Kappa = 0,69). A prevalência de HSV (96,7%), sífilis (6,4%) e HIV (3,2%) foi maior que a descrita na literatura, enquanto a prevalência de clamídia (6,4%) e gonorreia (1,6%) foi similar. O HPV foi detectado em 53,2% das mulheres, sendo 32,3% delas infectadas pelo HPV 16. No contexto das coinfecções, 38 mulheres (61,3%) tinham mais de uma IST. Conclusão: Visto que a maioria das mulheres analisadas era acometida por mais de uma IST, nossos resultados sugerem que uma triagem rotineira dessas infecções nas unidades de saúde poderia auxiliar na detecção precoce, bem como no tratamento e na prevenção. Essas medidas também impactariam no controle do câncer cervical das pacientes.

Reactivation of latent HIV-1 in vitro using an ethanolic extract from Euphorbia umbellata (Euphorbiaceae) latex.

Euphorbia umbellata (E. umbellata) belongs to Euphorbiaceae family, popularly known as Janauba, and its latex contains a combination of phorbol esters with biological activities described to different cellular protein kinase C (PKC) isoforms. Here, we identified deoxi-phorbol esters present in E. umbellata latex alcoholic extract that are able to increase HIV transcription and reactivate virus from latency models. This activity is probably mediated by NF-kB activation followed by nuclear translocation and binding to the HIV LTR promoter. In addition, E. umbellata latex extract induced the production of pro inflammatory cytokines in vitro in human PBMC cultures. This latex extract also activates latent virus in human PBMCs isolated from HIV positive patients as well as latent SIV in non-human primate primary CD4+ T lymphocytes. Together, these results indicate that the phorbol esters present in E. umbellata latex are promising candidate compounds for future clinical trials for shock and kill therapies to promote HIV cure and eradication.

Comparison of the editing patterns and editing efficiencies of TALEN and CRISPR-Cas9 when targeting the human CCR5 gene.

The human C-C chemokine receptor type-5 (CCR5) is the major transmembrane co-receptor that mediates HIV-1 entry into target CD4+ cells. Gene therapy to knock-out the CCR5 gene has shown encouraging results in providing a functional cure for HIV-1 infection. In gene therapy strategies, the initial region of the CCR5 gene is a hotspot for producing functional gene knock-out. Such target gene editing can be done using programmable endonucleases such as transcription activator-like effector nucleases (TALEN) or clustered regularly interspaced short palindromic repeats (CRISPR-Cas9). These two gene editing approaches are the most modern and effective tools for precise gene modification. However, little is known of potential differences in the efficiencies of TALEN and CRISPR-Cas9 for editing the beginning of the CCR5 gene. To examine which of these two methods is best for gene therapy, we compared the patterns and amount of editing at the beginning of the CCR5 gene using TALEN and CRISPR-Cas9 followed by DNA sequencing. This comparison revealed that CRISPR-Cas9 mediated the sorting of cells that contained 4.8 times more gene editing than TALEN+ transfected cells.

Comparison of the editing patterns and editing efficiencies of TALEN and CRISPR-Cas9 when targeting the human CCR5 gene

Abstract The human C-C chemokine receptor type-5 (CCR5) is the major transmembrane co-receptor that mediates HIV-1 entry into target CD4+ cells. Gene therapy to knock-out the CCR5 gene has shown encouraging results in providing a functional cure for HIV-1 infection. In gene therapy strategies, the initial region of the CCR5 gene is a hotspot for producing functional gene knock-out. Such target gene editing can be done using programmable endonucleases such as transcription activator-like effector nucleases (TALEN) or clustered regularly interspaced short palindromic repeats (CRISPR-Cas9). These two gene editing approaches are the most modern and effective tools for precise gene modification. However, little is known of potential differences in the efficiencies of TALEN and CRISPR-Cas9 for editing the beginning of the CCR5 gene. To examine which of these two methods is best for gene therapy, we compared the patterns and amount of editing at the beginning of the CCR5 gene using TALEN and CRISPR-Cas9 followed by DNA sequencing. This comparison revealed that CRISPR-Cas9 mediated the sorting of cells that contained 4.8 times more gene editing than TALEN+ transfected cells.

Zika Virus Infects, Activates, and Crosses Brain Microvascular Endothelial Cells, without Barrier Disruption.

Zika virus (ZIKV) has been associated to central nervous system (CNS) harm, and virus was detected in the brain and cerebrospinal fluids of microcephaly and meningoencephalitis cases. However, the mechanism by which the virus reaches the CNS is unclear. Here, we addressed the effects of ZIKV replication in human brain microvascular endothelial cells (HBMECs), as an in vitro model of blood brain barrier (BBB), and evaluated virus extravasation and BBB integrity in an in vivo mouse experimental model. HBMECs were productively infected by African and Brazilian ZIKV strains (ZIKVMR766 and ZIKVPE243), which induce increased production of type I and type III IFN, inflammatory cytokines and chemokines. Infection with ZIKVMR766 promoted earlier cellular death, in comparison to ZIKVPE243, but infection with either strain did not result in enhanced endothelial permeability. Despite the maintenance of endothelial integrity, infectious virus particles crossed the monolayer by endocytosis/exocytosis-dependent replication pathway or by transcytosis. Remarkably, both viruses' strains infected IFNAR deficient mice, with high viral load being detected in the brains, without BBB disruption, which was only detected at later time points after infection. These data suggest that ZIKV infects and activates endothelial cells, and might reach the CNS through basolateral release, transcytosis or transinfection processes. These findings further improve the current knowledge regarding ZIKV dissemination pathways.

Immune activation in amniotic fluid from Zika virus-associated microcephaly.

Recent advances in the understanding of neuropathogenesis associated with Zika virus (ZIKV) infection has led to descriptions of neonatal microcephaly cases. However, none of these reports have evaluated the humoral response during ZIKV infection. We report here polyfunctional immune activation associated with increased interferon-gamma-inducible protein 10, interleukin (IL)-6, IL-8, vascular endothelial growth factor (VEGF), monocyte chemoattractive protein 1 (MCP-1), and granulocyte colony-stimulating factor (G-CSF) levels in the amniotic fluid of ZIKV-positive pregnant women with neonatal microcephaly. These cytokines have been associated not only with neuronal damage, but also with differentiation and proliferation of neural progenitor cells. Our results suggested that the immune activation caused by ZIKV infection in the uterine environment could also interfere with fetal development. ANN NEUROL 2017;81:152-156.

Chloroquine, an Endocytosis Blocking Agent, Inhibits Zika Virus Infection in Different Cell Models.

Zika virus (ZIKV) infection in utero might lead to microcephaly and other congenital defects. Since no specific therapy is available thus far, there is an urgent need for the discovery of agents capable of inhibiting its viral replication and deleterious effects. Chloroquine is widely used as an antimalarial drug, anti-inflammatory agent, and it also shows antiviral activity against several viruses. Here we show that chloroquine exhibits antiviral activity against ZIKV in Vero cells, human brain microvascular endothelial cells, human neural stem cells, and mouse neurospheres. We demonstrate that chloroquine reduces the number of ZIKV-infected cells in vitro, and inhibits virus production and cell death promoted by ZIKV infection without cytotoxic effects. In addition, chloroquine treatment partially reveres morphological changes induced by ZIKV infection in mouse neurospheres.

Zika Virus Causing Encephalomyelitis Associated With Immunoactivation.

Brazil has experienced a Zika virus (ZIKV) outbreak with increased incidence of congenital malformations and neurological manifestations. We describe a case of a 26-year-old Brazilian Caucasian man infected with ZIKV and diagnosed with encephalomyelitis. Brain and spinal cord images showed hyperintense lesions on T2 and fluid-attenuated inversion recovery (FLAIR), and levels of proinflammatory cytokines in the cerebrospinal fluid showed a remarkable increase of interleukin (IL)-6 and IL-8. The observed pattern suggests immune activation during the acute phase, along with the neurological impairment, with normalization in the recovery phase. This is the first longitudinal report of ZIKV infection causing encephalomyelitis with documented immune activation.