Gene encoding a deubiquitinating enzyme is mutated in artesunate- and chloroquine-resistant rodent malaria parasites§

Artemisinin- and artesunate-resistant Plasmodium chabaudi mutants, AS-ART and AS-ATN, were previously selected from chloroquine-resistant clones AS-30CQ and AS-15CQ respectively. Now, a genetic cross between AS-ART and the artemisinin-sensitive clone AJ has been analysed by Linkage Group Selection. A genetic linkage group on chromosome 2 was selected under artemisinin treatment. Within this locus, we identified two different mutations in a gene encoding a deubiquitinating enzyme. A distinct mutation occurred in each of the clones AS-30CQ and AS-ATN, relative to their respective progenitors in the AS lineage. The mutations occurred independently in different clones under drug selection with chloroquine (high concentration) or artesunate. Each mutation maps to a critical residue in a homologous human deubiquitinating protein structure. Although one mutation could theoretically account for the resistance of AS-ATN to artemisinin derivates, the other cannot account solely for the resistance of AS-ART, relative to the responses of its sensitive progenitor AS-30CQ. Two lines of Plasmodium falciparum with decreased susceptibility to artemisinin were also selected. Their drug-response phenotype was not genetically stable. No mutations in the UBP-1 gene encoding the P. falciparum orthologue of the deubiquitinating enzyme were observed. The possible significance of these mutations in parasite responses to chloroquine or artemisinin is discussed

Identification of a mutant PfCRT-mediated chloroquine tolerance phenotype in Plasmodium falciparum.

International audienceMutant forms of the Plasmodium falciparum transporter PfCRT constitute the key determinant of parasite resistance to chloroquine (CQ), the former first-line antimalarial, and are ubiquitous to infections that fail CQ treatment. However, treatment can often be successful in individuals harboring mutant pfcrt alleles, raising questions about the role of host immunity or pharmacokinetics vs. the parasite genetic background in contributing to treatment outcomes. To examine whether the parasite genetic background dictates the degree of mutant pfcrt-mediated CQ resistance, we replaced the wild type pfcrt allele in three CQ-sensitive strains with mutant pfcrt of the 7G8 allelic type prevalent in South America, the Oceanic region and India. Recombinant clones exhibited strain-dependent CQ responses that ranged from high-level resistance to an incremental shift that did not meet CQ resistance criteria. Nonetheless, even in the most susceptible clones, 7G8 mutant pfcrt enabled parasites to tolerate CQ pressure and recrudesce in vitro after treatment with high concentrations of CQ. 7G8 mutant pfcrt was found to significantly impact parasite responses to other antimalarials used in artemisinin-based combination therapies, in a strain-dependent manner. We also report clinical isolates from French Guiana that harbor mutant pfcrt, identical or related to the 7G8 haplotype, and manifest a CQ tolerance phenotype. One isolate, H209, harbored a novel PfCRT C350R mutation and demonstrated reduced quinine and artemisinin susceptibility. Our data: 1) suggest that high-level CQR is a complex biological process dependent on the presence of mutant pfcrt; 2) implicate a role for variant pfcrt alleles in modulating parasite susceptibility to other clinically important antimalarials; and 3) uncover the existence of a phenotype of CQ tolerance in some strains harboring mutant pfcrt

Differential Drug Efflux or Accumulation Does Not Explain Variation in the Chloroquine Response of Plasmodium falciparum Strains Expressing the Same Isoform of Mutant PfCRT▿

Mutant forms of the Plasmodium falciparum chloroquine resistance transporter (PfCRT) mediate chloroquine resistance by effluxing the drug from the parasite's digestive vacuole, the acidic organelle in which chloroquine exerts its parasiticidal effect. However, different parasites bearing the same mutant form of PfCRT can vary substantially in their chloroquine susceptibility. Here, we have investigated the biochemical basis for the difference in chloroquine response among transfectant parasite lines having different genetic backgrounds but bearing the same mutant form of PfCRT. Despite showing significant differences in their chloroquine susceptibility, all lines with the mutant PfCRT showed a similar chloroquine-induced H+ leak from the digestive vacuole, indicative of similar rates of PfCRT-mediated chloroquine efflux. Furthermore, all lines showed similarly reduced levels of drug accumulation. Factors other than chloroquine efflux and accumulation therefore influence the susceptibility to this drug in parasites expressing mutant PfCRT. Furthermore, in some but not all strains bearing mutant PfCRT, the 50% inhibitory concentration (IC50) for chloroquine and the degree of resistance compared to that of recombinant control parasites varied with the length of the parasite growth assays. In these parasites, the 50% inhibitory concentration for chloroquine measured in 72- or 96-h assays was significantly lower than that measured in 48-h assays. This highlights the importance of considering the first- and second-cycle activities of chloroquine in future studies of parasite susceptibility to this drug

Cell-free DNA screening in clinical practice: abnormal autosomal aneuploidy and microdeletion results

BackgroundSince its commercial release in 2011 cell-free DNA screening has been rapidly adopted as a routine prenatal genetic test. However, little is known about its performance in actual clinical practice.ObjectiveWe sought to investigate factors associated with the accuracy of abnormal autosomal cell-free DNA results.Study designWe conducted a retrospective cohort study of 121 patients with abnormal cell-free DNA results from a referral maternal-fetal medicine practice from March 2013 through July 2015. Patients were included if cell-free DNA results for trisomy 21, trisomy 18, trisomy 13, or microdeletions (if reported by the laboratory) were positive or nonreportable. The primary outcome was confirmed aneuploidy or microarray abnormality on either prenatal or postnatal karyotype or microarray. Secondary outcomes were identifiable associations with in vitro fertilization, twins, ultrasound findings, testing platform, and testing laboratory. Kruskal-Wallis or Fisher exact tests were used as appropriate.ResultsA total of 121 patients had abnormal cell-free DNA results for trisomy 21, trisomy 18, trisomy 13, and/or microdeletions. In all, 105 patients had abnormal cell-free DNA results for trisomy 21, trisomy 18, and trisomy 13. Of these, 92 (87.6%) were positive and 13 (12.4%) were nonreportable. The results of the 92 positive cell-free DNA were for trisomy 21 (48, 52.2%), trisomy 18 (22, 23.9%), trisomy 13 (17, 18.5%), triploidy (2, 2.2%), and positive for >1 parameter (3, 3.3%). Overall, the positive predictive value of cell-free DNA was 73.5% (61/83; 95% confidence interval, 63-82%) for all trisomies (by chromosome: trisomy 21, 83.0% [39/47; 95% confidence interval, 69-92%], trisomy 18, 65.0% [13/20; 95% confidence interval, 41-84%], and trisomy 13, 43.8% [7/16; 95% confidence interval, 21-70%]). Abnormal cell-free DNA results were associated with positive serum screening (by group: trisomy 21 [17/48, 70.8%]; trisomy 18 [7/22, 77.8%]; trisomy 13 [3/17, 37.5%]; nonreportable [2/13, 16.7%]; P = .004), and abnormal first-trimester ultrasound (trisomy 21 [25/45, 55.6%]; trisomy 18 [13/20, 65%]; trisomy 13 [6/14, 42.9%]; nonreportable [1/13, 7.7%]; P = .003). There was no association between false-positive rates and testing platform, but there was a difference between the 4 laboratories (P = .018). In all, 26 patients had positive (n = 9) or nonreportable (n = 17) microdeletion results. Seven of 9 screens positive for microdeletions underwent confirmatory testing; all were false positives.ConclusionThe positive predictive value of 73.5% for cell-free DNA screening for autosomal aneuploidy is lower than reported. The positive predictive value for microdeletion testing was 0%. Diagnostic testing is needed to confirm abnormal cell-free DNA results for aneuploidy and microdeletions

From Mosquitos to Humans: Genetic Evolution of Zika Virus

Submitted by sandra infurna ([email protected]) on 2016-06-21T12:16:55Z No. of bitstreams: 1 mirna_bonaldo_etal_IOC_2016.pdf: 4687639 bytes, checksum: 6512110ad27726b93d2b67ceded4953c (MD5)Approved for entry into archive by sandra infurna ([email protected]) on 2016-06-21T13:27:37Z (GMT) No. of bitstreams: 1 mirna_bonaldo_etal_IOC_2016.pdf: 4687639 bytes, checksum: 6512110ad27726b93d2b67ceded4953c (MD5)Made available in DSpace on 2016-06-21T13:27:37Z (GMT). No. of bitstreams: 1 mirna_bonaldo_etal_IOC_2016.pdf: 4687639 bytes, checksum: 6512110ad27726b93d2b67ceded4953c (MD5) Previous issue date: 2016Submitted by Angelo Silva ([email protected]) on 2016-07-07T11:16:51Z No. of bitstreams: 3 mirna_bonaldo_etal_IOC_2016.pdf.txt: 54766 bytes, checksum: 49b7761745c8b9a9cd8e211c2eb498cb (MD5) mirna_bonaldo_etal_IOC_2016.pdf: 4687639 bytes, checksum: 6512110ad27726b93d2b67ceded4953c (MD5) license.txt: 2991 bytes, checksum: 5a560609d32a3863062d77ff32785d58 (MD5)Approved for entry into archive by sandra infurna ([email protected]) on 2016-07-07T12:16:07Z (GMT) No. of bitstreams: 3 license.txt: 2991 bytes, checksum: 5a560609d32a3863062d77ff32785d58 (MD5) mirna_bonaldo_etal_IOC_2016.pdf: 4687639 bytes, checksum: 6512110ad27726b93d2b67ceded4953c (MD5) mirna_bonaldo_etal_IOC_2016.pdf.txt: 54766 bytes, checksum: 49b7761745c8b9a9cd8e211c2eb498cb (MD5)Made available in DSpace on 2016-07-07T12:16:07Z (GMT). No. of bitstreams: 3 license.txt: 2991 bytes, checksum: 5a560609d32a3863062d77ff32785d58 (MD5) mirna_bonaldo_etal_IOC_2016.pdf: 4687639 bytes, checksum: 6512110ad27726b93d2b67ceded4953c (MD5) mirna_bonaldo_etal_IOC_2016.pdf.txt: 54766 bytes, checksum: 49b7761745c8b9a9cd8e211c2eb498cb (MD5) Previous issue date: 2016University of California. Department of Microbiology, Immunology and Molecular Genetics. Los Angeles, CA, USA / Chinese Academy of Medical Sciences & Peking Union Medical College. Institute of Basic Medical Sciences. Center for Systems Medicine. Beijing, China / Suzhou Institute of Systems Medicine, Suzhou, Jiangsu 215123, China.University of California. Department of Microbiology, Immunology and Molecular Genetics. Los Angeles, CA, USA / David Geffen School of Medicine at UCLA. Department of Obstetrics & Gynecology. Los Angeles, CA, USA.Chinese Academy of Medical Sciences & Peking Union Medical College. Institute of Basic Medical Sciences. Center for Systems Medicine. Beijing, China / Suzhou Institute of Systems Medicine, Suzhou, Jiangsu 215123, China.University of California. Department of Microbiology, Immunology and Molecular Genetics. Los Angeles, CA, USA / Chinese Academy of Sciences. Institute of Biophysics. Beijing 100101, China.University of California. Department of Microbiology, Immunology and Molecular Genetics. Los Angeles, CA, USA.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratorio Biologia Molecular de Flavivírus. Rio de Janeiro, RJ, Brasil.University of California. 8UCLA Center for World Health,. Los Angeles, CA, USA.David Geffen School of Medicine at UCLA. Division of Pediatric Infectious Diseases. Los Angeles, CA, USA.Chinese Academy of Medical Sciences & Peking Union Medical College. Institute of Basic Medical Sciences. Center for Systems Medicine. Beijing, China / Suzhou Institute of Systems Medicine, Suzhou, Jiangsu 215123, China.University of California. Department of Microbiology, Immunology and Molecular Genetics. Los Angeles, CA, USA .University of California. Department of Microbiology, Immunology and Molecular Genetics. Los Angeles, CA, USA / Chinese Academy of Medical Sciences & Peking Union Medical College. Institute of Basic Medical Sciences. Center for Systems Medicine. Beijing, China / Suzhou Institute of Systems Medicine, Suzhou, Jiangsu 215123, China.Initially isolated in 1947, Zika virus (ZIKV) has recently emerged as a significant public health concern. Sequence analysis of all 41 known ZIKV RNA open reading frames to date indicates that ZIKV has undergone significant changes in both protein and nucleotide sequences during the past half century

Zika virus infection in pregnant women in Rio de Janeiro: preliminary report

Artigo liberado em acesso aberto como parte do acordo para tornar público todos os dados produzidos sobre o vírus zika - Compartilhamento de dados em emergências de saúde pública - http://www.wellcome.ac.uk/News/Media-office/Press-releases/2016/WTP060169.htmVersão final do artigo - handle https://www.arca.fiocruz.br/handle/icict/17780Submitted by Claudete Queiroz ([email protected]) on 2016-03-08T19:16:29Z No. of bitstreams: 1 Zika Virus Infection in Pregnant Women - preliminary report.pdf: 646105 bytes, checksum: 76b9427d455f2a5cbb1aa17b53e4cfc6 (MD5)Approved for entry into archive by Claudete Queiroz ([email protected]) on 2016-03-09T11:54:02Z (GMT) No. of bitstreams: 1 Zika Virus Infection in Pregnant Women - preliminary report.pdf: 646105 bytes, checksum: 76b9427d455f2a5cbb1aa17b53e4cfc6 (MD5)Made available in DSpace on 2016-03-09T11:54:02Z (GMT). No. of bitstreams: 1 Zika Virus Infection in Pregnant Women - preliminary report.pdf: 646105 bytes, checksum: 76b9427d455f2a5cbb1aa17b53e4cfc6 (MD5) Previous issue date: 2016Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Biomedical Research Institute of Southern California. Oceanside, California, EUA.Fundação Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Saúde da Mulher, da Criança e do Adolescente Fernandes Figueira. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Saúde da Mulher, da Criança e do Adolescente Fernandes Figueira. Rio de Janeiro, RJ, Brasil.David Geffen UCLA School of Medicine, Los Angeles, EUA.David Geffen UCLA School of Medicine, Los Angeles, EUA.David Geffen UCLA School of Medicine, Los Angeles, EUA.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.David Geffen UCLA School of Medicine, Los Angeles, EUA.BACKGROUND Zika virus (ZIKV) has been linked to neonatal microcephaly. To characterize the spectrum of ZIKV disease in pregnancy, we followed patients in Rio de Janeiro to describe clinical manifestations in mothers and repercussions of acute ZIKV infection in fetuses. METHODS We enrolled pregnant women in whom a rash had developed within the previous 5 days and tested blood and urine specimens for ZIKV by reverse-transcriptase–polymerasechain-reaction assays. We followed the women prospectively and collected clinical and ultrasonographic data. RESULTS A total of 88 women were enrolled from September 2015 through February 2016; of these 88 women, 72 (82%) tested positive for ZIKV in blood, urine, or both. The timing of acute ZIKV infection ranged from 5 to 38 weeks of gestation. Predominant clinical features included pruritic descending macular or maculopapular rash, arthralgias, conjunctival injection, and headache; 28% had fever (short-term and low-grade).Women who were positive for ZIKV were more likely than those who were negative for the virus to have maculopapular rash (44% vs. 12%, P=0.02), conjunctival involvement (58% vs. 13%, P=0.002), and lymphadenopathy (40% vs. 7%, P=0.02). Fetal ultrasonography was performed in 42 ZIKV-positive women (58%) and in all ZIKV-negative women. Fetal abnormalities were detected by Doppler ultrasonography in 12 of the 42 ZIKV-positive women (29%) and in none of the 16 ZIKV-negative women. Adverse findings included fetal deaths at 36 and 38 weeks of gestation (2 fetuses), in utero growth restriction with or without microcephaly (5 fetuses), ventricular calcifications or other central nervous system (CNS) lesions (7 fetuses), and abnormal amniotic fluid volume or cerebral or umbilical artery flow (7 fetuses). To date, 8 of the 42 women in whom fetal ultrasonography was performed have delivered their babies, and the ultrasonographic findings have been confirmed. CONCLUSIONS Despite mild clinical symptoms, ZIKV infection during pregnancy appears to be associated with grave outcomes, including fetal death, placental insufficiency, fetal growth restriction, and CNS injury

Zika virus infection in pregnant women in Rio de Janeiro

Artigo liberado em acesso aberto como parte do acordo para tornar público todos os dados produzidos sobre o vírus zika - Compartilhamento de dados em emergências de saúde pública - http://www.wellcome.ac.uk/News/Media-office/Press-releases/2016/WTP060169.htmVersão preliminar - handle https://www.arca.fiocruz.br/handle/icict/13063Submitted by Claudete Fernandes ([email protected]) on 2017-02-09T16:51:52Z No. of bitstreams: 1 Zika Virus Infection in Pregnant Women in Rio de Janeiro.pdf: 469292 bytes, checksum: 1690c1bd4e969661795a93985558eb03 (MD5)Approved for entry into archive by Claudete Fernandes ([email protected]) on 2017-02-09T17:54:26Z (GMT) No. of bitstreams: 1 Zika Virus Infection in Pregnant Women in Rio de Janeiro.pdf: 469292 bytes, checksum: 1690c1bd4e969661795a93985558eb03 (MD5)Made available in DSpace on 2017-02-09T17:54:26Z (GMT). No. of bitstreams: 1 Zika Virus Infection in Pregnant Women in Rio de Janeiro.pdf: 469292 bytes, checksum: 1690c1bd4e969661795a93985558eb03 (MD5) Previous issue date: 2016Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.David Geffen UCLA School of Medicine, Los Angeles, EUA.David Geffen UCLA School of Medicine, Los Angeles, EUA.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.David Geffen UCLA School of Medicine, Los Angeles, EUA.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Biomedical Research Institute of Southern California. Oceanside, California, EUA.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Saúde da Mulher, da Criança e do Adolescente Fernandes Figueira. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Saúde da Mulher, da Criança e do Adolescente Fernandes Figueira. Rio de Janeiro, RJ, Brasil.Universidade de São Paulo. Faculdade de Medicina. São Paulo, SP.Karolinska Institutet, Stockholm.Medical University of Graz, Graz, Austria.David Geffen UCLA School of Medicine, Los Angeles, EUA.David Geffen UCLA School of Medicine, Los Angeles, EUA.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.David Geffen UCLA School of Medicine, Los Angeles, EUA.BACKGROUND: Zika virus (ZIKV) has been linked to central nervous system malformations in fetuses. To characterize the spectrum of ZIKV disease in pregnant women and infants, we followed patients in Rio de Janeiro to describe clinical manifestations in mothers and repercussions of acute ZIKV infection in infants. METHODS: We enrolled pregnant women in whom a rash had developed within the previous 5 days and tested blood and urine specimens for ZIKV by reverse-transcriptase–polymerasechain-reaction assays. We followed women prospectively to obtain data on pregnancy and infant outcomes. RESULTS: A total of 345 women were enrolled from September 2015 through May 2016; of these, 182 women (53%) tested positive for ZIKV in blood, urine, or both. The timing of acute ZIKV infection ranged from 6 to 39 weeks of gestation. Predominant maternal clinical features included a pruritic descending macular or maculopapular rash, arthralgias, conjunctival injection, and headache; 27% had fever (short-term and low-grade). By July 2016, a total of 134 ZIKV-affected pregnancies and 73 ZIKV-unaffected pregnancies had reached completion, with outcomes known for 125 ZIKV-affected and 61 ZIKV-unaffected pregnancies. Infection with chikungunya virus was identified in 42% of women without ZIKV infection versus 3% of women with ZIKV infection (P<0.001). Rates of fetal death were 7% in both groups; overall adverse outcomes were 46% among offspring of ZIKV-positive women versus 11.5% among offspring of ZIKV-negative women (P<0.001). Among 117 live infants born to 116 ZIKV-positive women, 42% were found to have grossly abnormal clinical or brain imaging findings or both, including 4 infants with microcephaly. Adverse outcomes were noted regardless of the trimester during which the women were infected with ZIKV (55% of pregnancies had adverse outcomes after maternal infection in the first trimester, 52% after infection in the second trimester, and 29% after infection in the third trimester). CONCLUSIONS: Despite mild clinical symptoms in the mother, ZIKV infection during pregnancy is deleterious to the fetus and is associated with fetal death, fetal growth restriction, and a spectrum of central nervous system abnormalities. (Funded by Ministério da Saúde do Brasil and others.)