Panoramic snapshot of serum soluble mediator interplay in pregnant women with convalescent COVID-19: an exploratory study

IntroductionSARS-CoV-2 infection during pregnancy can induce changes in the maternal immune response, with effects on pregnancy outcome and offspring. This is a cross-sectional observational study designed to characterize the immunological status of pregnant women with convalescent COVID-19 at distinct pregnancy trimesters. The study focused on providing a clear snapshot of the interplay among serum soluble mediators.MethodsA sample of 141 pregnant women from all prenatal periods (1st, 2nd and 3rd trimesters) comprised patients with convalescent SARS-CoV-2 infection at 3-20 weeks after symptoms onset (COVID, n=89) and a control group of pre-pandemic non-infected pregnant women (HC, n=52). Chemokine, pro-inflammatory/regulatory cytokine and growth factor levels were quantified by a high-throughput microbeads array.ResultsIn the HC group, most serum soluble mediators progressively decreased towards the 2nd and 3rd trimesters of pregnancy, while higher chemokine, cytokine and growth factor levels were observed in the COVID patient group. Serum soluble mediator signatures and heatmap analysis pointed out that the major increase observed in the COVID group related to pro-inflammatory cytokines (IL-6, TNF-α, IL-12, IFN-γ and IL-17). A larger set of biomarkers displayed an increased COVID/HC ratio towards the 2nd (3x increase) and the 3rd (3x to 15x increase) trimesters. Integrative network analysis demonstrated that HC pregnancy evolves with decreasing connectivity between pairs of serum soluble mediators towards the 3rd trimester. Although the COVID group exhibited a similar profile, the number of connections was remarkably lower throughout the pregnancy. Meanwhile, IL-1Ra, IL-10 and GM-CSF presented a preserved number of correlations (≥5 strong correlations in HC and COVID), IL-17, FGF-basic and VEGF lost connectivity throughout the pregnancy. IL-6 and CXCL8 were included in a set of acquired attributes, named COVID-selective (≥5 strong correlations in COVID and <5 in HC) observed at the 3rd pregnancy trimester.Discussion and conclusionFrom an overall perspective, a pronounced increase in serum levels of soluble mediators with decreased network interplay between them demonstrated an imbalanced immune response in convalescent COVID-19 infection during pregnancy that may contribute to the management of, or indeed recovery from, late complications in the post-symptomatic phase of the SARS-CoV-2 infection in pregnant women

A new index to discriminate between iron deficiency anemia and thalassemia trait.

Background: The most common microcytic and hypochromic anemias are iron deficiencyanemia and thalassemia trait. Several indices to discriminate iron deficiency anemia fromthalassemia trait have been proposed as simple diagnostic tools. However, some of the bestdiscriminative indices use parameters in the formulas that are only measured in moderncounters and are not always available in small laboratories.The development of an index with good diagnostic accuracy based only on parametersderived from the blood cell count obtained using simple counters would be useful in theclinical routine. Thus, the aim of this study was to develop and validate a discriminativeindex to differentiate iron deficiency anemia from thalassemia trait. Methods: To develop and to validate the new formula, blood count data from 106 (thalassemiatrait: 23 and iron deficiency: 83) and 185 patients (thalassemia trait: 30 and iron deficiency:155) were used, respectively. Iron deficiency, _-thalassemia trait and _-thalassemia trait wereconfirmed by gold standard tests (low serum ferritin for iron deficiency anemia, HbA2> 3.5%for _-thalassemia trait and using molecular biology for the _-thalassemia trait).Results: The sensitivity, specificity, efficiency, Youden?s Index, area under receiver operatingcharacteristic curve and Kappa coefficient of the new formula, called the Matos & CarvalhoIndex were 99.3%, 76.7%, 95.7%, 76.0, 0.95 and 0.83, respectively. Conclusion: The performance of this index was excellent with the advantage of being solelydependent on the mean corpuscular hemoglobin concentration and red blood cell countobtained from simple automatic counters and thus may be of great value in underdevelopedand developing countries

Severe preeclampsia: association of genes polymorphisms and maternal cytokines production in Brazilian population

Submitted by Nuzia Santos ([email protected]) on 2016-01-07T15:59:11Z No. of bitstreams: 1 Severe preeclampsia_ Association of genes polymorphisms and maternal cytokines production in Brazilian population - 1-s2.0-S104.pdf: 4325596 bytes, checksum: 251b5d9d3999c73b8d95da9078481995 (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2016-01-07T16:03:16Z (GMT) No. of bitstreams: 1 Severe preeclampsia_ Association of genes polymorphisms and maternal cytokines production in Brazilian population - 1-s2.0-S104.pdf: 4325596 bytes, checksum: 251b5d9d3999c73b8d95da9078481995 (MD5)Made available in DSpace on 2016-01-07T16:03:16Z (GMT). No. of bitstreams: 1 Severe preeclampsia_ Association of genes polymorphisms and maternal cytokines production in Brazilian population - 1-s2.0-S104.pdf: 4325596 bytes, checksum: 251b5d9d3999c73b8d95da9078481995 (MD5) Previous issue date: 2015Universidade Federal de Minas Gerais. Faculdade de Farmácia. Departamento de Análises Clínicas e Toxicológicas. Belo Horizonte, MG, Brasil/Universidade Federal de São João Del Rei. Faculdade de Medicina. Divinópolis, MG, BrasilUniversidade Federal de Minas Gerais. Faculdade de Farmácia. Departamento de Análises Clínicas e Toxicológicas. Belo Horizonte, MG, BrasilSimile Instituto de Imunologia Aplicada. Belo Horizonte, MG, BrasilSimile Instituto de Imunologia Aplicada. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Faculdade de Farmácia. Departamento de Análises Clínicas e Toxicológicas. Belo Horizonte, MG, BrasilFundação Osvaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Biomarcadores de Diagnóstico e Monitoração. Belo Horizonte, MG, BrasilFundação Osvaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Biomarcadores de Diagnóstico e Monitoração. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Faculdade de Farmácia. Departamento de Análises Clínicas e Toxicológicas. Belo Horizonte, MG, BrasilIntroduction: Preeclampsia (PE) is a multi-system disorder of pregnancy characterized by hypertension and proteinuria. Healthy pregnancy is associated with a controlled inflammatory process, which is exacerbated in PE in response to excessive placental stimuli. Gene expression levels can affect inflammation and immune regulation. It is known that differences in cytokine allele frequencies amongst populations may contribute to difference in the incidence of several diseases. Objective: The aim of this study was to investigate the frequency of TNF-α, IL-6, IFN-γ and IL-10 genes polymorphisms and their relationship with the cytokines plasma levels in PE. Methods: A total of 281 women were included in this study; 116 with severe PE, 107 normotensive pregnant and 58 non-pregnant women. Cytokine genotyping was carried out by the polymerase chain reaction. The analyzed polymorphisms were: TNF-α (−308 G → A), IL-10 (−1082 G → A), IL-6 (−174 G → C), and IFN-γ (+874 A → T). Cytokine plasma levels were measured by Cytometric Bead Array method. Results: A higher frequency of the IFN-γ (+874) T/T genotype in severe PE comparing to normotensive pregnant women was found (P < 0.001). TNF-α, IL-6 and IFN-γ plasma levels were higher in PE women compared to non-pregnant women (P < 0.001; P < 0.001; P = 0.004). IL-6 and IFN-γ levels were also higher in PE women compared to normotensive pregnant (P < 0.001; P = 0.010). IL-10 levels were higher in normotensive pregnant women compared to PE (P < 0.001). IFN-γ and IL-6 genes polymorphisms influenced the genic expression in PE and normotensive pregnant women, respectively. Conclusions: These results suggest that IFN-γ seems to play a role in PE occurrence

Longitudinal assessment of D-dimer and plasminogen activator inhibitor type-1 plasma levels in pregnant women with risk factors for preeclampsia

Objective: Investigating D-Dimer/D-Di and plasminogen activator inhibitor type-1/PAI-1 levels throughout gestation in women with preeclampsia/PE risk factors. Methods: D-Di and PAI-1 plasma levels were determined in 28 women at 12–19, 20–29, 30–34 and 35–40 weeks of gestation. Results: D-Di was lower at 12–19 weeks and higher at 30–34 weeks in women who developed PE versus who did not develop it. D-Di increased throughout gestation in both groups, peaking earlier in pregnant women who developed PE versus who did not develop it. PA1-1 increased across gestation, but it didn’t differ between groups. Conclusion: D-Di was able to discriminate these groups of women at 12–19 and 30–34 weeks of gestation.</p

Association among ACE, ESR1 polymorphisms and preeclampsia in Brazilian pregnant women.

Background: Genetic, immune and environmental factors are involved in preeclampsia (PE) etiopathogenesis. Considering that hypertension and poor placental perfusion are important features in PE, polymorphisms in the angiotensin-converting enzyme (ACE) and estrogen nuclear receptor 1 (ESR1) genes could be involved in the predisposition and/or development of the disease. The aim of this study was to evaluate if polymorphisms in ACE and ESR1 genes were associated with PE occurrence. Material and Methods: This case-control study included 209 Brazilian pregnant women (107 with severe PE and 102 normotensive controls). The polymorphisms were investigated by polymerase chain reaction (PCR) followed by polyacrylamide gel electrophoresis. Results: No significant difference between PE versus normotensive pregnant women, as well as early versus late PE, was observed when compared the allelic and genotypic frequencies of insertion/deletion polymorphism in intron 16 of the ACE gene and the single nucleotide polymorphisms (SNPs - rs2234693 and rs9340799) of the ESR1 gene. Conclusion: This pioneer study involving Brazilian women showed no association among the studied polymorphisms and PE, which suggests that ins/del ACE and SNPs ESR1 do not contribute to this disease occurrence in Brazil

Flow cytometry reticulocyte counting using acridine orange: validation of a new protocol

Submitted by Nuzia Santos ([email protected]) on 2015-03-26T13:19:34Z No. of bitstreams: 1 2014_170.pdf: 266255 bytes, checksum: 231d8aaac025e76936cafbd975f83c8c (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-03-26T13:19:49Z (GMT) No. of bitstreams: 1 2014_170.pdf: 266255 bytes, checksum: 231d8aaac025e76936cafbd975f83c8c (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-03-26T13:26:11Z (GMT) No. of bitstreams: 1 2014_170.pdf: 266255 bytes, checksum: 231d8aaac025e76936cafbd975f83c8c (MD5)Made available in DSpace on 2015-03-26T13:26:11Z (GMT). No. of bitstreams: 1 2014_170.pdf: 266255 bytes, checksum: 231d8aaac025e76936cafbd975f83c8c (MD5) Previous issue date: 2014Universidade Federal de Minas Gerais. Faculdade de Farmácia. Belo Horizonte, MG, Brasil/Prefeitura de Belo Horizonte. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Faculdade de Farmácia. Departamento de Análises Clinicas e Toxicologicas. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Faculdade de Farmácia. Departamento de Análises Clinicas e Toxicologicas. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Faculdade de Farmácia. Belo Horizonte, MG, BrasilCentro Universitário Newton Paiva. Belo Horizonte, MG, BrasilLaboratório de Patologia Clínica São Paulo. Serviço de Citometria. Belo Horizonte, MG, BrasilFundação Hemominas. Hematologia. Belo Horizonte, MG, BrasilLaboratório de Patologia Clínica São Paulo. Hematologia Clinica. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Rene Rachou. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Rene Rachou. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Rene Rachou. Belo Horizonte, MG, BrasilIntrodução: Atualmente, a contagem de reticulócitos representa um desafio para os laboratórios clínicos no Brasil, principalmente os de pequeno e médio porte, nos quais ainda se utiliza o método manual. Este método apresenta algumas limitações, classificando-se como tedioso, demorado e de baixa precisão. Objetivos: O presente estudo desenvolveu e avaliou o desempenho de um novo protocolo laboratorial para contagem de reticulócitos por citometria de fluxo (CF) utilizando acridine orange (AO) como corante, visando padronizar um protocolo preciso, de fácil e rápida execução e custo acessível. Após a padronização do novo protocolo desenvolvido (CF/AO), fez-se a comparação com o método manual. Os resultados foram analisados de acordo com recomendações do National Committee for Clinical Laboratory Standards (NCCLS), atualmente Clinical and Laboratory Standards Institute (CLSI), para avaliar a intercambialidade entre os métodos, por meio da análise de regressão linear e teste t pareado, além de outros testes de controle de qualidade. Conclusão: Diante dos resultados obtidos referentes à correlação entre os métodos e os testes voltados ao controle de qualidade, pode-se admitir que o CF/AO estabelecido para contagem de reticulócitos possui vantagens inegáveis quando comparado com o método manual.Introduction: Currently, the reticulocyte counting is a challenge for clinical laboratories in Brazil, mainly for the ordinary ones, which still use the manual method. This method has some limitations, since it consists of a laborious method, time consuming, with low accuracy. Objectives: This study has developed and evaluated the performance of a New Laboratory Protocol for flow cytometry (FC) reticulocytes counting using acridine orange (AO) as dye, aiming to standardize a more precise, easy, fast implementation, and low cost protocol. After standardization of the New Protocol (FC/AO), it was compared with the manual method. The results were analyzed according to the recommendations of the National Committee for Clinical Laboratory Standards (NCCLS), now known as Clinical and Laboratory Standards Institute (CLSI), to evaluate the interchangeability of methods in linear regression analysis and paired t test, besides other quality control tests. Conclusion: Based on these results concerning to the correlation between the methods and the tests related to quality control, we can admit that FC/AO for reticulocyte counting shows undeniable advantages when compared to the preexisting manual method