Oxidative stress assessment in sickle cell anemia patients treated with hydroxyurea.

Hydroxyurea (HU) is used as a therapy in sickle cell anemia (SCA). Many studies have established that HU improves patient quality of life by reducing symptoms. However, the effect of HU on erythrocytes is not well-described. We evaluated several parameters related to oxidative stress and total lipid content of erythrocytes in patients with SCA. The patient cohort consisted of 7 SCA patients treated with HU, 17 untreated SCA patients, and 15 healthy subjects. Erythrocytes from patients with SCA displayed increased oxidative stress relative to the control group, including higher thiobarbituric acid reactive substances (TBARS), Fe3+ content, and osmotic fragility, and decreased total cholesterol. We observed that treatment of SCA patients with HU increased Fe3+ content and activity of glutathione peroxidase, and decreased glutathione reductase activity, glutathione levels, total cholesterol, and phospholipid content comaperaded to patients untreated with HU. Thus, HU alters biochemical characteristics of erythrocytes; future studies will determine whether they are beneficial or not.

Aspartate aminotransferase to platelet ratio index (APRI) for differentiation of primary and secondary infection by dengue virus / Aplicação do índice de relação aspartato aminotransferase sobre plaquetas (APRI) na diferenciação de infecção primária e secundária pelo vírus da dengue

ABSTRACT INTRODUCTION: Dengue virus (DENV) infection has been considered a major public health problem in tropical countries. The unavailability of serologic testing in public health centers might adversely impact patients' outcome. OBJECTIVE: This study aimed to evaluate the accuracy of mean platelet volume (MPV) and aspartate aminotransferase (AST) to platelet ratio index (APRI) as laboratory markers of DENV infection that could be used to differentiate primary and secondary infections. METHODS: We assessed laboratory results from 503 patients with positive rapid test for DENV infection. RESULTS: Severe thrombocytopenia and increased liver involvement were observed in patients with DENV heterotypic secondary infection. Our data suggest that APRI was able to distinguish patients with primary and secondary infection (p = 0.006) with a relevant sensitivity (75%), specificity (76%) and a cut-off of 1.06. A total of 80 out of 105 (76%) patients with primary DENV infection had APRI ≤ 1.06, and 12 (75%) with secondary DENV infection had APRI > 1.06. On the other hand, MPV did not show significance in the differentiation of types of infection, coming up with poor area under the receiver operating characteristic (ROC) curve (0.61). CONCLUSION: APRI seems to be a powerful tool for early identification of DENV secondary infection cases in health centers.

RESUMO INTRODUÇÃO: A infecção pelo vírus da dengue (DENV) é considerada um grande problema de saúde pública nos países tropicais. A indisponibilidade de testes sorológicos em centros de saúde pública pode afetar negativamente o prognóstico do paciente. OBJETIVO: Este estudo teve como objetivo avaliar a precisão do volume médio de plaquetas (MPV) e o índice da relação de aspartato aminotransferase (AST) sobre plaquetas (APRI) como marcadores laboratoriais de infecção por DENV, que poderiam ser utilizados para diferenciar infecções primárias e secundárias. MÉTODOS: Foram avaliados os resultados laboratoriais de 503 pacientes com teste rápido positivo para infecção por DENV. RESULTADOS: Foram observadas trombocitopenia grave e disfunção hepática em pacientes com infecção secundária heterogênea por DENV. Nossos dados sugerem que o APRI foi capaz de distinguir os pacientes com infecção primária e secundária (p = 0, 006), com relevante sensibilidade (75%) e especificidade (76%) e corte de 1, 06. Um total de 80 de 105 (76%) pacientes com infecção primária por DENV tinha APRI ≤ 1, 06; e 12 (75%) com infecção secundária por DENV, APRI > 1, 06. Por outro lado, o MPV não mostrou significância na diferenciação de tipos de infecção, apresentando baixo valor da área sob a curva de característica de operação do receptor (ROC) (0, 61). CONCLUSÃO: APRI parece ser uma ferramenta poderosa para identificação precoce de casos de infecção secundária de DENV em centros de saúde.

Evaluation of lipid profile, high-sensitivity C-reactive protein and D-dimer in users of oral contraceptives of different types / Avaliação de perfil lipídico, proteína C reativa ultrassensível e dímero D de usuárias de diferentes tipos de contraceptivos orais

ABSTRACT Introduction: The use of oral contraceptives increases women's risk of developing cardiovascular and thromboembolic diseases, due to alterations in hemostatic and lipid profile. Objectives: Analyze the association between the use of different types of oral contraceptives with lipid profile and levels of serum high-sensitivity C-reactive protein (hsCRP) and plasma D-dimer. Methods: One hundred fifty-four participants were divided into the following groups: control nonusers (n = 41), medium-dose users (n= 32), third-generation low-dose users (n = 40), and fourth-generation low-dose users (n = 41). Triglycerides and total cholesterol serum levels were determined by colorimetric enzymatic method; high-density lipoprotein (HDL) cholesterol levels, by precipitation method; low-density lipoprotein (LDL) cholesterol levels, by Friedewald equation; hsCRP levels, by immunoturbidimetric method; and D-dimer levels, by fluorescence immunoassay. Results: Oral contraceptive users had higher serum levels of triglycerides, total cholesterol, HDL cholesterol (HDL-C), HDL/LDL index and hsCRP compared to controls. Medium-dose users had higher D-dimer plasma levels than controls and higher triglycerides serum levels than low-dose users. Triglycerides, hsCRP and D-dimer were positively correlated to each other. Conclusion: The use of combined oral contraceptives was associated with an unfavorable lipid profile and a chronic subclinical inflammation, with atherogenic potential. Furthermore, medium-dose contraceptives induced a higher thrombogenic potential, since they were associated with increased D-dimer levels in comparison to low-dose ones.

RESUMO Introdução: O uso de anticoncepcionais orais aumenta o risco de desenvolvimento de doenças cardiovasculares e tromboembólicas devido a alterações no perfil lipídico e hemostático. Objetivo: Analisar a associação entre o uso de diferentes tipos de anticoncepcionais orais com o perfil lipídico e os níveis da proteína C reativa ultrassensível (PCRus) e do dímero D. Métodos: Cento e quarenta e cinco participantes foram divididas em: não usuárias (n = 41), usuárias de média dose (n = 32), usuárias de terceira geração de baixa dose (n = 40) e usuárias de quarta geração de baixa dose (n = 41). Níveis de triglicerídeos e colesterol total foram determinados pelo método enzimático colorimétrico; colesterol da lipoproteína de alta densidade (HDL), pelo método de precipitação; colesterol da lipoproteína de baixa densidade (LDL), pela equação de Friedewald; PCRus, por imunoturbidimetria; e dímero D, por imunoensaio fluorescente. Resultados: As usuárias de anticoncepcionais orais apresentaram maiores níveis de triglicerídeos, colesterol total, HDL, índice HDL/LDL e PCRus do que as não usuárias. As usuárias de anticoncepcionais de média dose apresentaram maiores níveis de dímero D do que as não usuárias, e maiores níveis de triglicerídeos do que as usuárias de anticoncepcionais de baixa dose. Triglicerídeos, PCRus e dímero D apresentaram correlação positiva uns com os outros. Conclusão: O uso de anticoncepcionais orais combinados está associado ao perfil lipídico desfavorável e à inflamação crônica subclínica, com potencial aterogênico. Além disso, os anticoncepcionais orais de média dose induziram maior potencial trombogênico, já que foram relacionados com níveis maiores de dímero D em comparação com os de baixa dose.

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

INTRODUCTION: 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.

Soluble endoglin, transforming growth factor-Beta 1 and soluble tumor necrosis factor alpha receptors in different clinical manifestations of preeclampsia.

BACKGROUND: Despite intensive research, the etiopathogenesis of preeclampsia (PE) remains uncertain. Inflammatory and angiogenic factors are thought to play considerable roles in this disease. The objective of this study was to investigate the association between soluble endoglin (sEng), transforming growth factor beta-1 (TGF-ß1) and tumor necrosis factor alpha soluble receptors (sTNF-Rs) and the clinical manifestations of PE. METHODS: Plasma levels of sEng, TGF-ß1 and sTNF-Rs were determined by ELISA in 23 non-pregnant, 21 normotensive pregnant and 43 PE women. PE women were stratified into subgroups according to the severity [mild (n = 12) and severe (n = 31)] and onset-time of the disease [early (n = 19) and late (n = 24)]. RESULTS: Pregnancy was associated with higher levels of sEng, sTNF-R1 and sTNF-R2 than the non-pregnant state. Moreover, PE women had higher levels of sEng and sTNF-R1 than normotensive pregnant women. No difference was found in TGF-ß1 levels, comparing the three study groups. Late PE had higher levels of sTNF-R1 and sTNF-R2 than early PE. No significant differences were found in sEng and TGF-ß1 comparing early and late PE. sEng levels were higher in severe PE than in mild PE and no difference was found for TGF-ß1, sTNF-R1 and sTNF-R2 levels. There was a positive correlation among sEng, TNF-R1 and sTNF-2 levels. Logistic regression analysis revealed that primiparity and sEng levels are independently associated with the development of PE. Furthermore, sEng levels are independently associated with the disease severity. CONCLUSIONS: These results suggest that pregnancy is a condition associated with higher levels of anti-angiogenic and pro-inflammatory factors than the non-pregnant state and that PE is associated with an imbalance of these factors in the maternal circulation.

Severe preeclampsia: are hemostatic and inflammatory parameters associated?

BACKGROUND: Preeclampsia (PE) is characterized by hypertension and proteinuria. A predisposition to endothelial dysfunction, which may trigger abnormal activation of the hemostatic and/or inflammatory systems, is thought to play a crucial part in pathogenesis of PE. We investigated the relationship between hemostatic and inflammatory parameters in women with severe PE. METHODS: D-Dimer, PAI-1, IL-8, IL-6, TNF-α, and IFN-γ concentrations were measured in 59 pregnant women with severe PE (sPE), 49 normotensive pregnant and 48 non-pregnant women. RESULTS: D-Dimer and PAI-1 were higher in women with sPE compared to normotensive pregnant and non-pregnant women. IL-8, IL-6, and IFN-γ also were higher in women with sPE compared to normotensive pregnant women. However, only IL-6 and IFN-γ were higher in women with sPE compared to non-pregnant women. Moreover, D-Dimer and PAI-1 showed an elevated area under ROC curve proving to be excellent for discriminating sPE. Correlation analysis showed a weak correlation between D-Dimer and IL-8 and between PAI-1 and IFN-γ in sPE. CONCLUSION: D-Di and PAI-1 concentrations showed to be an important tool for monitoring sPE.

Severe preeclampsia goes along with a cytokine network disturbance towards a systemic inflammatory state.

Although preeclampsia causes high maternal/fetal morbidity and mortality, the etiology of this multi-system disorder still remains to be elucidated. Herein, we have characterized the cytokine plasma levels in severe preeclamptic women compared to normotensive pregnant and non-pregnant women, aiming to better understand the immunological network and its clinical significance for the pathogenesis and severity of preeclampsia. A total of 219 women were selected. The study population was composed of three groups referred as severe preeclamptic, normotensive pregnant and non-pregnant women. Cytokine plasma levels were determined using commercially available kits, Cytometric Beads Array - CBA to quantify TNF-α, IFN-γ, IL-4, IL-5, IL-10, IL-1ß, IL-6, IL-8 and IL-12. Our findings demonstrated that severe preeclamptic state is associated with high levels of pro-inflammatory cytokines IL-8, IL-6, and IFN-γ (P < 0.05 for all) whereas normotensive pregnancy evolves high levels of regulatory cytokine IL-10 (P < 0.05). Moreover, an outstanding pro-inflammatory "cytokine signature" could be observed in severe preeclamptic women display, while an overall regulatory state is the hallmark for normotensive pregnancy. In summary, our data showed that elevated levels of pro-inflammatory cytokines in the maternal circulation with a deviation in the "IL-8 × IL-6" axis towards IFN-γ might drive the cytokine network in preeclamptic women towards an excessive systemic inflammatory state.

CD4-CD8-αß and γδ T cells display inflammatory and regulatory potentials during human tuberculosis.

T-cells play an important role controlling immunity against pathogens and therefore influence the outcome of human diseases. Although most T-lymphocytes co-express either CD4 or CD8, a smaller T-cell subset found the in the human peripheral blood that expresses the αß or γδ T-cell-receptor (TCR) lacks the CD4 and CD8 co-receptors. These double negative (DN) T-cells have been shown to display important immunological functions in human diseases. To better understand the role of DN T-cells in human Mycobacterium tuberculosis, we have characterized their frequency, activation and cytokine profile in a well-defined group of tuberculosis patients, categorized as severe and non-severe based on their clinical status. Our data showed that whereas high frequency of αß DN T-cells observed in M. tuberculosis-infected patients are associated with disease severity, decreased proportion of γδ DN T-cells are found in patients with severe tuberculosis. Together with activation of CD4(+) and CD8(+) T-cells, higher frequencies of both αß and γδ DN T-cells from tuberculosis patients also express the chronic activation marker HLA-DR. However, the expression of CD69, an early activation marker, is selectively observed in DN T-cells. Interestingly, while αß and γδ DN T-cells from patients with non-severe tuberculosis display a pro-inflammatory cytokine profile, characterized by enhanced IFN-γ, the γδ DN T-cells from patients with severe disease express a modulatory profile exemplified by enhanced interleukin-10 production. Overall, our findings suggest that αß and γδ DN T-cell present disparate immunoregulatory potentials and seems to contribute to the development/maintenance of distinct clinical aspects of TB, as part of the complex immunological network triggered by the TB infection.

Pre-eclampsia: relationship between coagulation, fibrinolysis and inflammation.

Pre-eclampsia (PE) is a multi-system disorder of human pregnancy, characterised by hypertension and proteinuria. Although the pathogenesis of PE is not fully understood, predisposition to endothelial dysfunction is thought to play a crucial part. Despite intensive research there is no reliable test for screening purposes or to inform decision making towards effective treatment for PE. Understanding the link between PE, abnormal haemostatic activation and inflammation may help to elucidate some of the patho-physiology of the disease; primary preventative measures and targeted therapies at an early stage of the disease could then be considered. In the present paper we discuss potential causal links between PE, haemostasis and inflammation. The potential implications of such interaction on the pathogenesis of PE are also addressed.