Complement System Activation Is a Plasma Biomarker Signature during Malaria in Pregnancy.

Malaria in pregnancy (MiP) is a public health problem in malaria-endemic areas, contributing to detrimental outcomes for both mother and fetus. Primigravida and second-time mothers are most affected by severe anemia complications and babies with low birth weight compared to multigravida women. Infected erythrocytes (IE) reach the placenta, activating the immune response by placental monocyte infiltration and inflammation. However, specific markers of MiP result in poor outcomes, such as low birth weight, and intrauterine growth restriction for babies and maternal anemia in women infected with Plasmodium falciparum are limited. In this study, we identified the plasma proteome signature of a mouse model infected with Plasmodium berghei ANKA and pregnant women infected with Plasmodium falciparum infection using quantitative mass spectrometry-based proteomics. A total of 279 and 249 proteins were quantified in murine and human plasma samples, of which 28% and 30% were regulated proteins, respectively. Most of the regulated proteins in both organisms are involved in complement system activation during malaria in pregnancy. CBA anaphylatoxin assay confirmed the complement system activation by the increase in C3a and C4a anaphylatoxins in the infected plasma compared to non-infected plasma. Moreover, correlation analysis showed the association between complement system activation and reduced head circumference in newborns from Pf-infected mothers. The data obtained in this study highlight the correlation between the complement system and immune and newborn outcomes resulting from malaria in pregnancy.

Virulence Profiles of Wild-Type, P.1 and Delta SARS-CoV-2 Variants in K18-hACE2 Transgenic Mice.

Since December 2019, the world has been experiencing the COVID-19 pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and we now face the emergence of several variants. We aimed to assess the differences between the wild-type (Wt) (Wuhan) strain and the P.1 (Gamma) and Delta variants using infected K18-hACE2 mice. The clinical manifestations, behavior, virus load, pulmonary capacity, and histopathological alterations were analyzed. The P.1-infected mice showed weight loss and more severe clinical manifestations of COVID-19 than the Wt and Delta-infected mice. The respiratory capacity was reduced in the P.1-infected mice compared to the other groups. Pulmonary histological findings demonstrated that a more aggressive disease was generated by the P.1 and Delta variants compared to the Wt strain of the virus. The quantification of the SARS-CoV-2 viral copies varied greatly among the infected mice although it was higher in P.1-infected mice on the day of death. Our data revealed that K18-hACE2 mice infected with the P.1 variant develop a more severe infectious disease than those infected with the other variants, despite the significant heterogeneity among the mice.

Genetic diversity of Plasmodium vivax isolates from pregnant women in the Western Brazilian Amazon: a prospective cohort study.

Background: Each year, 92 million pregnant women are at risk of contracting malaria during pregnancy, with the underestimation of the mortality and morbidity burden associated with Plasmodium vivax. During pregnancy, P. vivax infection is associated with low birth weight, maternal anaemia, premature delivery, and stillbirth. In the State of Acre (Brazil), high transmission leaves pregnant women at greater risk of contracting malaria and having a greater number of recurrences. The study of genetic diversity and the association of haplotypes with adverse pregnancy effects is of great importance for the control of the disease. Here we investigate the genetic diversity of P. vivax parasites infecting pregnant women across their pregnancies. Methods: P. vivax DNA was extracted from 330 samples from 177 women followed during pregnancy, collected in the State of Acre, Brazil. All samples were negative for Plasmodium falciparum DNA. Sequence data for the Pvmsp1 gene was analysed alongside data from six microsatellite (MS) markers. Allelic frequencies, haplotype frequencies, expected heterozygosity (HE) were calculated. Whole genome sequencing (WGS) was conducted on four samples from pregnant women and phylogenetic analysis performed with other samples from South American regions. Findings: Initially, the pregnant women were stratified into two groups-1 recurrence and 2 or more recurrences-in which no differences were observed in clinical gestational outcomes or in placental histological changes between the two groups. Then we evaluated the parasites genetically. An average of 18.5 distinct alleles were found at each of the MS loci, and the HE calculated for each marker indicates a high genetic diversity occurring within the population. There was a high percentage of polyclonal infections (61.7%, 108/175), and one haplotype (H1) occurred frequently (20%), with only 9 of the haplotypes appearing in more than one patient. Interpretation: Most pregnant women had polyclonal infections that could be the result of relapses and/or re-infections. The high percentage of H1 parasites, along with the low frequency of many other haplotypes are suggestive of a clonal expansion. Phylogenetic analysis shows that P. vivax population within pregnant women clustered with other Brazilian samples in the region. Funding: FAPESP and CNPq - Brazil.

Mapping and characterizing areas with high levels of malaria in pregnancy in Brazil: A spatiotemporal analysis.

Background: Malaria in pregnancy (MiP) is a public health problem in the Brazilian Amazon region that requires special attention due to associated serious adverse consequences, such as low birth weight, increased prematurity and spontaneous abortion rates. In Brazil, there have been no comprehensive epidemiological studies of MiP. In this study, we aimed to explore the spatial and spatiotemporal patterns of MiP in Brazil and epidemiologically characterize this population of pregnant women over a period of 15 years. Methods: We performed a national-scale ecological analysis using a Bayesian space-time hierarchical model to estimate the incidence rates of MiP between 1 January 2004 and 31 December 2018. We mapped the high-incidence clusters among pregnant women aged 10-49 years old using a Poisson model, and we characterized the population based on data from the Epidemiological Surveillance Information System for Malaria (SIVEP-Malaria). Findings: We consolidated the data of 61,833 women with MiP in Brazil. Our results showed a reduction of 50·1% (95% CI: 47·3 to 52·9) in the number of malaria cases over the period analysed, with Plasmodium vivax malaria having the highest incidence. MiP was widely distributed throughout the Amazon region, and spatial and spatiotemporal analyses revealed statistically significant clusters in some municipalities of Amazonas, Acre, Rondônia and Pará. In addition, we observed that younger pregnant women had a higher risk of infection, and the administration of appropriate treatment requires more attention. Interpretation: This nationwide study provides robust evidence that, despite the reduction in the number of MiP cases in the country, it remains a serious public health problem, especially for young pregnant women. Our analyses highlight focus areas for strengthening interventions to control and eliminate MiP. Funding: FAPESP and CNPq - Brazil.

TLR4-Endothelin Axis Controls Syncytiotrophoblast Motility and Confers Fetal Protection in Placental Malaria.

Pregnancy-associated malaria is often associated with adverse pregnancy outcomes. Placental circulatory impairments are an intriguing and unsolved component of malaria pathophysiology. Here, we uncovered a Toll-like receptor 4 (TLR4)-TRIF-endothelin axis that controls trophoblast motility and is linked to fetal protection during Plasmodium infection. In a cohort of 401 pregnancies from northern Brazil, we found that infection during pregnancy reduced expression of endothelin receptor B in syncytiotrophoblasts, while endothelin expression was only affected during acute infection. We further show that quantitative expression of placental endothelin and endothelin receptor B proteins are differentially controlled by maternal and fetal TLR4 alleles. Using murine malaria models, we identified placental autonomous responses to malaria infection mediated by fetally encoded TLR4 that not only controlled placental endothelin gene expression but also correlated with fetal viability protection. In vitro assays showed that control of endothelin expression in fetal syncytiotrophoblasts exposed to Plasmodium-infected erythrocytes was dependent on TLR4 via the TRIF pathway but not MyD88 signaling. Time-lapse microscopy in syncytiotrophoblast primary cultures and cell invasion assays demonstrated that ablation of TLR4 or endothelin receptor blockade abrogates trophoblast collective motility and cell migration responses to infected erythrocytes. These results cohesively substantiate the hypothesis that fetal innate immune sensing, namely, the TRL4-TRIF pathway, exerts a fetal protective role during malaria infection by mediating syncytiotrophoblast vasoregulatory responses that counteract placental insufficiency.

Adverse pregnancy outcomes are associated with Plasmodium vivax malaria in a prospective cohort of women from the Brazilian Amazon.

BACKGROUND: Malaria in Brazil represents one of the highest percentages of Latin America cases, where approximately 84% of infections are attributed to Plasmodium (P.) vivax. Despite the high incidence, many aspects of gestational malaria resulting from P. vivax infections remain poorly studied. As such, we aimed to evaluate the consequences of P. vivax infections during gestation on the health of mothers and their neonates in an endemic area of the Amazon. METHODS AND FINDINGS: We have conducted an observational cohort study in Brazilian Amazon between January 2013 and April 2015. 600 pregnant women were enrolled and followed until delivery. After applying exclusion criteria, 329 mother-child pairs were included in the analysis. Clinical data regarding maternal infection, newborn's anthropometric measures, placental histopathological characteristics, and angiogenic and inflammatory factors were evaluated. The presence of plasma IgG against the P. vivax (Pv) MSP119 protein was used as marker of exposure and possible associations with pregnancy outcomes were analyzed. Multivariate logistic regression analysis revealed that P. vivax infections during the first trimester of pregnancy are associated with adverse gestational outcomes such as premature birth (adjusted odds ratio [aOR] 8.12, 95% confidence interval [95%CI] 2.69-24.54, p < 0.0001) and reduced head circumference (aOR 3.58, 95%CI 1.29-9.97, p = 0.01). Histopathology analysis showed marked differences between placentas from P. vivax-infected and non-infected pregnant women, especially regarding placental monocytes infiltrate. Placental levels of vasomodulatory factors such as angiopoietin-2 (ANG-2) and complement proteins such as C5a were also altered at delivery. Plasma levels of anti-PvMSP119 IgG in infected pregnant women were shown to be a reliable exposure marker; yet, with no association with improved pregnancy outcomes. CONCLUSIONS: This study indicates that P. vivax malaria during the first trimester of pregnancy represents a higher likelihood of subsequent poor pregnancy outcomes associated with marked placental histologic modification and angiogenic/inflammatory imbalance. Additionally, our findings support the idea that antibodies against PvMSP119 are not protective against poor pregnancy outcomes induced by P. vivax infections.

Plasmodium falciparum infection dysregulates placental autophagy.

Plasmodium (P.) falciparum malaria during pregnancy has been frequently associated with severe consequences such as maternal anemia, abortion, premature birth, and reduced birth weight. Placental damage promotes disruption of the local homeostasis; though, the mechanisms underlying these events are still to be elucidated. Autophagy is a fundamental homeostatic mechanism in the natural course of pregnancy by which cells self-recycle in order to survive in stressful environments. Placentas from non-infected and P. falciparum-infected women during pregnancy were selected from a previous prospective cohort study conducted in the Brazilian Amazon (Acre, Brazil). Newborns from infected women experienced reduced birth weight (P = 0.0098) and placental immunopathology markers such as monocyte infiltrate (P < 0.0001) and IL-10 production (P = 0.0122). The placentas were evaluated for autophagy-related molecules. As a result, we observed reduced mRNA levels of ULK1 (P = 0.0255), BECN1 (P = 0.0019), and MAP1LC3B (P = 0.0086) genes in placentas from P. falciparum-infected, which was more striking in those diagnosed with placental malaria. Despite the protein levels of these genes followed the same pattern, the observed reduction was not statistically significant in placentas from P. falciparum-infected women. Nevertheless, our data suggest that chronic placental immunopathology due to P. falciparum infection leads to autophagy dysregulation, which might impair local homeostasis during malaria in pregnancy that may result in poor pregnancy outcomes.

Association of Malaria Infection During Pregnancy With Head Circumference of Newborns in the Brazilian Amazon.

Importance: Malaria during pregnancy is associated with adverse events for the fetus and newborn, but the association of malaria during pregnancy with the head circumference of the newborn is unclear. Objective: To investigate the association of malaria during pregnancy with fetal head growth. Design, Setting, and Participants: Two cohort studies were conducted at the general maternity hospital of Cruzeiro do Sul (Acre, Brazil) in the Amazonian region. One cohort study prospectively enrolled noninfected and malaria-infected pregnant women who were followed up until delivery, between January 2013 and April 2015. The other cohort study was assembled retrospectively using clinical and malaria data from all deliveries that occurred between January 2012 and December 2013. Data analyses were conducted from January to August 2017 and revised in November 2018. Clinical data from pregnant women and anthropometric measures of their newborns were evaluated. A total of 600 pregnant women were enrolled through volunteer sampling (prospective cohort study), and 4697 pregnant women were selected by population-based sampling (retrospective cohort study). After application of exclusion criteria, data from 251 (prospective cohort study) and 232 (retrospective cohort study) malaria-infected and 158 (prospective cohort study) and 3650 (retrospective cohort study) noninfected women were evaluated. Exposure: Malaria during pregnancy. Main Outcomes and Measures: The primary end point was the incidence of altered head circumference in newborns delivered from malaria-infected mothers compared with that from noninfected mothers. Secondary end points included measures of placental pathology relative to newborn head circumference. Results: In total, 4291 maternal-child pairs were analyzed. Among 409 newborns in the prospective cohort study, the mothers of 251 newborns had malaria during pregnancy, infected with Plasmodium vivax, Plasmodium falciparum, or both. Among 3882 newborns in the retrospective cohort study, 232 were born from mothers that had malaria during pregnancy. The prevalence of newborns with a small head (19 [30.7%] in the prospective cohort study and 30 [36.6%] in the retrospective cohort study) and the prevalence of microcephaly among newborns (5 [8.1%] in the prospective cohort study and 6 [7.3%] in the retrospective cohort study) were higher among newborns from women infected with P falciparum during pregnancy. Multivariate logistic regression analyses revealed that P falciparum infection during pregnancy represented a significant risk factor for the occurrence of small head circumference in newborns (prospective cohort study: odds ratio, 3.15; 95% CI, 1.52-6.53; P = .002; retrospective cohort study: odds ratio, 1.91; 95% CI, 1.21-3.04; P = .006). Placental pathologic findings corroborated this association, with more syncytial nuclear aggregates and inflammatory infiltrates occurring in placentas of newborns born with decreased head circumference. Conclusions and Relevance: This study indicates that falciparum malaria during pregnancy is associated with decreased head circumference in newborns, which is in turn associated with evidence of placental malaria.

Malaria during pregnancy and newborn outcome in an unstable transmission area in Brazil: A population-based record linkage study.

BACKGROUND: Malaria in pregnancy (MiP) is one of the major causes of mortality and morbidity in tropical regions, causing maternal anemia, intrauterine growth retardation, preterm birth, and low birth weight (LBW). The integration of the information systems on pregnancy and malaria could prove to be a useful method of improved decision making for better maternal-child health. METHODS: A population-based observational study acquired information retrospectively from all live births that occurred between 2006 and 2014 in Cruzeiro do Sul (Acre, Brazil). Social and clinical data of the mother and newborn was extracted from the Information System of Live Births. Malaria episodes information was obtained from the Brazilian Epidemiological Surveillance Information System Malaria. A deterministic record linkage was performed to assess malaria impact on pregnancy. RESULTS: The studied population presented a malaria incidence of 8.9% (1283 pregnant women infected), of which 63.9% infected by Plasmodium (P.) vivax. Reduction of newborn birth weight at term (small for gestational age (SGA) and LBW) has been found associated with P. vivax infection during pregnancy (SGA-OR 1.24, 95% CI 1.02-1.52, p = 0.035; term LBW-OR 1.39, 95% CI 1.03-1.88, p = 0.033). Additionally, P. falciparum infection during pregnancy has been found to be associated with preterm births (OR 1.54, 95% CI 1.09-2.18, p = 0.016), which is related with late preterm births (OR 1.59, 95% CI 1.11-2.27, p = 0.011). CONCLUSIONS: Despite the decrease of malaria cases during the evaluation period and regardless of Plasmodium species, we present evidence of the deleterious effects of MiP in a low transmission area in the Amazonian region.

Effectiveness of the Live Births Information System in the Far-Western Brazilian Amazon.

The Live Birth Information System (SINASC) was implemented in 1990 for the purpose of providing information about the live-birth characteristics for the establishment of specific health indicators. This work evaluates the information quality of SINASC in relation to its data completeness and coverage for five municipalities from the State of Acre from 2005 to 2010. Lack of information (not filled out or stated as "unknown") was estimated for each variable. Coverage was estimated comparing the Civil Register office statistics in accordance with the mother's municipality of residence. An increase in incompleteness of the majority of variables was observed, and also a decrease in coverage between 2005 and 2010 in these municipalities. These findings do not tally with results from the majority of studies that use SINASC as a data source. The results of this work highlight the relevance of continuous capacity building and the incentive for accurate and complete data inclusion, as well as awareness of the importance of SINASC for public health policies.

Effectiveness of the Live Births Information System in the Far-Western Brazilian Amazon / Eficácia do Sistema de Informação de Nascidos Vivos na Amazônia Extremo-Ocidental Brasileira

The Live Birth Information System (SINASC) was implemented in 1990 for the purpose of providing information about the live-birth characteristics for the establishment of specific health indicators. This work evaluates the information quality of SINASC in relation to its data completeness and coverage for five municipalities from the State of Acre from 2005 to 2010. Lack of information (not filled out or stated as "unknown") was estimated for each variable. Coverage was estimated comparing the Civil Register office statistics in accordance with the mother's municipality of residence. An increase in incompleteness of the majority of variables was observed, and also a decrease in coverage between 2005 and 2010 in these municipalities. These findings do not tally with results from the majority of studies that use SINASC as a data source. The results of this work highlight the relevance of continuous capacity building and the incentive for accurate and complete data inclusion, as well as awareness of the importance of SINASC for public health policies.

O Sistema de Informação de Nascidos Vivos (SINASC) foi implantado no ano de 1990 com o objetivo de fornecer dados sobre as características de nascidos vivos para o estabelecimento de indicadores de saúde específicos. Objetivo: O presente trabalho avalia a qualidade da informação do SINASC quanto à incompletude dos seus dados e da cobertura para cinco municípios do estado do Acre nos anos de 2005 e 2010. Métodos: Foi calculada a incompletude (definida como dados em branco/ignorado) de cada variável, assim como a cobertura desse sistema através da comparação com as estatísticas do Registro Civil, segundo município de residência da mãe. Resultados: Observou-se um aumento da incompletude da maioria das variáveis e uma diminuição da cobertura de 2005 para 2010 no conjunto dos municípios avaliados, destoando dos resultados obtidos na maioria dos estudos que utilizam o SINASC como fonte de dados. Conclusões: Os resultados deste trabalho apontam para a importância da contínua capacitação e também para o incentivo ao preenchimento dos dados de forma correta e completa, bem como a conscientização da importância do SINASC para as políticas públicas de saúde.

Atuação do enfermeiro na prescrição de contraceptivos hormonais na rede de atenção primária em saúde / Performance of nurses in prescribing hormonal contraceptives in the primary health care network / Acción de enfermeros en la prescripción de anticonceptivos hormonales en la red de atención primaria de salud

O estudo teve como objetivo conhecer e analisar a atuação do enfermeiro na prescrição dos contraceptivos hormonais reversíveis na Rede de Atenção Primária a Saúde. Trata-se de um estudo transversal e descritivo, tendo como sujeitos 64 enfermeiros lotados nas unidades assistenciais no período de setembro a novembro de 2010 no município de Rio Branco-Acre. Os dados foram coletados mediante entrevistas estruturadas utilizando um questionário. Os resultados evidenciaram que 96,9% dos enfermeiros prescrevem os métodos anticoncepcionais e que 90,6% tem conhecimento da legislação que rege a prescrição de enfermagem. Foi observado que a escolha do método contraceptivo pelos enfermeiros baseia-se na escolha da cliente e anamnese (36% e 34% respectivamente), e que 90% sempre orientavam quanto às vantagens e desvantagens de cada método. Assim, para escolher um método contraceptivo de forma livre e informada, os métodos devem estar disponíveis e devem ser dispensados por profissionais capacitados, após orientação correta e completa.

The aim of this study was to evaluate the performance of nurses in the prescription of hormonal reversible contraceptives in Primary Care. A descriptive and transversal study was conducted, with 64 nurses at healthcare units as subjects, in the period of September-November 2010 in the county of Rio Branco-Acre. Data were collected through structured interviews using a questionnaire. The results showed that 96.9% of nurses are prescribing hormonal contraceptives and that the majority (90.6%) is aware of the government laws about nursing prescription. We also observed that the contraceptive was chosen based on client's decision and anamnesis (36% and 34% respectively), and that 90% of the nurses always give orientation about advantages and disadvantages of each contraceptive method. Thus, for choosing a contraceptive method based on given information and free will, the methods must be available and dispensed by trained professionals, after correct and complete orientation.

El estudio tuvo como objetivo identificar y analizar el desempeño de los enfermeros en la prescripción de anticonceptivos hormonales reversibles en la Red de Atención Primaria. Se trata de un estudio descriptivo y transversal, que tiene como sujetos 64 enfermeros inscritos en las unidades de cuidado de salud en el período de septiembre a noviembre de 2010 en la ciudad de Rio Branco, Acre. Los datos fueron recolectados a través de entrevistas con un cuestionario estructurado. Los resultados mostraron que 96,9% de las enfermeras recetan los anticonceptivos y que 90,6% tiene conocimiento de las leyes que regulan la prescripción de enfermería. Se observó que la elección del método anticonceptivo por las enfermeras es basada en la decisión del cliente y la anamnesis (36% y 34% respectivamente) y que 90% siempre aconsejaban a respecto de las ventajas y desventajas de cada método. Así que para elegir un método anticonceptivo de manera libre e informada, todos los métodos deben estar disponible y ser dispensados por profesionales capacitados, después de la orientación correcta y completa.

[Performance of nurses in prescribing hormonal contraceptives in the primary health care network]. / Atuação do enfermeiro na prescrição de contraceptivos hormonais na rede de atenção primária em saúde.

The aim of this study was to evaluate the performance of nurses in the prescription of hormonal reversible contraceptives in Primary Care. A descriptive and transversal study was conducted, with 64 nurses at healthcare units as subjects, in the period of September-November 2010 in the county of Rio Branco-Acre. Data were collected through structured interviews using a questionnaire. The results showed that 96.9% of nurses are prescribing hormonal contraceptives and that the majority (90.6%) is aware of the government laws about nursing prescription. We also observed that the contraceptive was chosen based on client's decision and anamnesis (36% and 34% respectively), and that 90% of the nurses always give orientation about advantages and disadvantages of each contraceptive method. Thus, for choosing a contraceptive method based on given information and free will, the methods must be available and dispensed by trained professionals, after correct and complete orientation.