Cristina Catarino

Inflammatory Disturbances in Preeclampsia: Relationship between Maternal and Umbilical Cord Blood

Preeclampsia (PE) is one of the main causes of maternal and fetal mortality and morbidity. PE is associated with an inflammatory state and with oxidative stress, in maternal circulation. Our aim was to evaluate and compare the levels of oxidative stress and inflammatory markers in maternal and umbilical cord blood (UCB), in normal and PE pregnancies. We measured acute-phase proteins (CRP and α1-antitrypsin), proinflammatory cytokines (IL-6 and TNF-α), leukocyte activation (elastase, lactoferrin, sL-selectin, sVCAM, sPECAM), total antioxidant status (TAS), thiobarbituric acid reactive substances (TBARS), and uric acid levels. We studied 42 healthy pregnant women, 46 PE women, and their neonates. The concentrations of IL-6, TNF-α, α1-antitrypsin, CRP, sVCAM, uric acid, and TBARS were significantly higher, and sL-selectin was significantly lower in PE pregnant women as compared with normotensive pregnant women. In newborns uric acid, α1-antitrypsin, and CRP values were significantly higher in PE; leukocyte count, sL-selectin, lactoferrin, and the ratio elastase/α1-antitrypsin were significantly lower. Our data suggest that PE pregnancy is associated with an enhanced maternal inflammatory condition, which is reflected in fetal circulation. This enhanced inflammatory state seems to be related to endothelial dysfunction and increased cytokine synthesis, rather than with neutrophil activation.

Erythrocyte damage and leukocyte activation in ischemic stroke

BACKGROUND The traditional lipid risk factors can only predict some of the cardiovascular events. Our work has focused on new potential biological markers of risk, namely leukocyte activation and erythrocyte membrane damage, in ischemic stroke cases. METHODS Besides the traditional lipid profile, we evaluated the plasma levels of elastase and lactoferrin as markers of leukocyte activation, and membrane band 3 protein profile and membrane bound hemoglobin as markers of erythrocyte damage. Total and differential leukocyte counts and erythrocyte counts, hematocrit and hemoglobin concentrations were also evaluated. The lipid study included the evaluation of triglycerides, total cholesterol, high-density lipoprotein cholesterol (HDLc), low-density lipoprotein cholesterol (LDLc), apolipoprotein AI (Apo AI) and B (Apo B), and lipoprotein (a) (Lp(a)). The work was performed in a control group (n=29) with no history of cardiovascular events, presenting normal hematological and lipid values, and in a pathologic group (n=21) of ischemic stroke cases diagnosed by computed tomographic imaging. RESULTS We found that ischemic stroke was associated with significantly higher values of leukocytes, which seem to be activated, as shown by significant higher levels of elastase and lactoferrin. This activation seems to impose erythrocyte damage, as suggested by a significant increase in membrane bound hemoglobin and by a different band 3 profile. CONCLUSIONS Our data suggest that plasma levels of elastase and lactoferrin, together with levels of erythrocyte membrane bound hemoglobin and band 3 profile, could be used as powerful new markers of risk for cardiovascular events.

The role of adipocytes in the modulation of iron metabolism in obesity

A tight relationship between iron deficiency and obesity is known to exist. The chronic low‐grade inflammation that characterizes obesity enhances hepcidin production, the principal regulator of iron availability. Adipose tissue is known to secret interleukin‐6 and leptin that triggers hepcidin production. It was found that adipose tissue also expresses hepcidin and hemojuvelin, a regulator of hepcidin production. These recent findings suggest that adipose tissue may have an important role in erythropoiesis particularly on obesity that is still poorly clarified. This paper discusses these findings and how they can modulate erythropoiesis.

Fetal and maternal angiogenic/anti-angiogenic factors in normal and preeclamptic pregnancy

Few studies evaluated angiogenic/anti-angiogenic factors and endothelial (dys)function in both maternal and umbilical cord blood (UCB) in preeclampsia (PE). We aimed to clarify the role of placental growth factor (PlGF), vascular endothelial growth factor (VEGF), soluble vascular endothelial growth factor receptor 1 (VEGFR-1) and tissue plasminogen activator (tPA), by evaluating them in maternal and UCB in 42 normal and 46 preeclamptic (PEc) cases. In PE, maternal and UCB PlGF were significantly lower; maternal VEGF, sVEGFR-1 and tPA were significantly higher. In UCB, sVEGFR-1 and tPA were significantly higher in PEc cases, while VEGF and PlGF were significantly lower. A significant correlation between maternal and UCB sVEGFR-1, and between sVEGFR-1 and tPA both in maternal and UCB, was observed in PEc cases. In maternal and UCB circulation in PE, a close interaction seems to exist between endothelial dysfunction and angiogenesis disturbance, and sVEGFR-1 seems to play a central role in those disturbances.

Fetal lipoprotein changes in pre-eclampsia

Objective. To evaluate the impact of maternal lipid changes upon the fetus in pre‐eclampsia (PE) by evaluating lipid profile simultaneously in maternal and umbilical cord blood (UCB). Design. Case‐control study performed on healthy and pre‐eclamptic pregnant women and their neonates. Setting. The Department of Obstetrics and Gynecology, Hospital S. Joao and Faculty of Pharmacy, Porto, Portugal. Samples. Forty‐two healthy pregnancies and 46 pregnancies complicated with PE. Methods. Total cholesterol (TChol), HDL‐cholesterol (HDLc), LDL‐cholesterol (LDLc) and triglycerides (TG) levels were determined using enzymatic methods. Apolipoprotein (apo) A‐I, apoB and lipoprotein (a) [Lp(a)] values were measured by immunoturbidimetry. Main outcome measures. Fetal and maternal plasma levels of TChol, HDLc, LDLc, TG, apoA‐I, apoB and Lp(a). Results. Pre‐eclamptic women presented significantly higher values for TChol, LDLc, HDLc, TG, apoA‐I and apoB compared to normal pregnant women. In the UCB from pre‐eclamptic pregnancies, we observed significantly lower values for HDLc and apoA‐I, and significantly higher TG concentrations and LDLc/HDLc ratio when compared to normal cases. A positive correlation was observed between maternal TG levels and proteinuria, a marker of PE severity (r =0.40, p <0.01). Conclusions. Our data suggest that pre‐eclamptic pregnancy is associated with an enhanced hyperlipidemia, which seems to have a negative impact on fetal lipid profile, as reflected by a higher atherogenic LDLc/HDLc ratio and higher TG levels. These children, born of women with PE, may deserve a closer clinical follow‐up later in life.

Protein deficiency balance as a predictor of clinical outcome in hereditary spherocytosis

Abstract:  Vertical and horizontal interactions between membrane constituents account for integrity, strength and deformability of the erythrocyte. Disruption of vertical interactions caused by membrane protein deficiencies in hereditary spherocytosis (HS), favor membrane vesiculation with development of spherocytic cells. Our aim was to evaluate the hematological and clinical presentation of HS according to the type and amount of protein deficiency. We studied 81 Portuguese individuals, 71 belonging to 21 families plus 10 unrelated subjects, and found that 51 of them were HS patients. Patients were classified as presenting mild, typical or severe HS, according to laboratory results and clinical follow‐up. We performed screening tests and the standardized electrophoretic membrane protein analysis to identify and quantify protein deficiencies. We found band 3 and ankyrin deficiencies as the major causes for HS. The ratios between the value of the primary and/or secondary protein deficiencies showed significantly different values according to the severity of HS, and a significant inverse correlation with the severity of HS was observed. In mild HS, the ratios between protein deficiencies reflected equivalent protein deficiencies, while an unbalance was observed in typical HS, which was enhanced in severe HS. Our data suggest that the relative quantification of each major membrane protein and of the ratios between the values of protein deficiencies may be helpful in providing additional data about the clinical outcome of HS.

Band 3 as a marker of erythrocyte changes in pregnancy

Abstract:  Modifications in the erythrocyte membrane protein band 3 seem to mark the cell for death. A decrease in band 3 high molecular weight aggregates (HMWAg) and a rise in its proteolytic fragments (Pfrag) were described for younger erythrocytes. The aim of this work was to study the band 3 profile as a marker of erythrocyte changes in pregnancy and postpartum. We performed a cross‐sectional study in non‐pregnant controls (n = 24), in women in the first (n = 64), second (n = 48) and third (n = 67) trimesters of gestation, and also in the puerperium (24–48 h after delivery; n = 32); we also carried out a longitudinal study (n = 23) during the three trimesters of normal pregnancy. We evaluated the band 3 profile (% of band 3 monomer, HMWAg, and Pfrag) and the membrane‐bound haemoglobin. Total serum bilirrubin, glutathione peroxidase activity, red blood cell (RBC) count, haematocrit (Ht), haemoglobin (Hb) concentration, the haematimetric indices, and red cell distribution width were also evaluated. Similar results were found in pregnancy in both the cross‐sectional and longitudinal studies. We found that the RBC count, Hb, and Ht decreased significantly in pregnancy and in puerperium. Band 3 profile in the first trimester of pregnancy, when compared with controls, presented significantly reduced HMWAg and increased Pfrag. Comparing the first with the third trimester, we found a significant reduction in band 3 and a significant rise in Pfrag. However, between these same periods, HMWAg did not decrease. Our data suggest band 3 profile as a marker of erythrocyte changes in pregnancy, which are independent of the ‘physiological anaemia’ of pregnancy. These changes suggest an increase in damaged RBCs, but also an increase in younger RBCs in the maternal circulation.

Linkage of cytosolic peroxiredoxin 2 to erythrocyte membrane imposed by hydrogen peroxide-induced oxidative stress

Human erythrocyte peroxiredoxin 2 (Prx2) is a typical 2-cys cytosolic peroxiredoxin with thiol-dependent hydrogen peroxide scavenger activity. In a previous work, we reported Prx2 erythrocyte membrane linkage in some Hereditary Spherocytosis patients and that it seemed to be related to oxidative stress. The aim of the present work was to determine if Prx2 linkage to erythrocyte membrane could be induced by oxidative stress mediated by H(2)O(2) and to further understand how and why this process occurs. We performed in vitro assays in which catalase or both Hb autoxidation and catalase were inhibited, under H(2)O(2)-induced oxidative stress conditions. Erythrocyte membrane linked Prx2 was detected by immunoblotting and quantified by densitometry. As oxidative stress markers, we determined membrane bound hemoglobin and lipid peroxidation, and we found that their values increased with H(2)O(2) concentration. Prx2 linkage to the membrane also rose with increasing H(2)O(2) concentration, and was only observed when the oxidized form of the enzyme was present in the cytosol. Oxidized Hb and Prx2 membrane linkages appear to be independent processes, although, both result from oxidative stress and may be useful as oxidative stress and/or erythrocyte damage/senescence markers.

Relationship between maternal and cord blood hemostatic disturbances in preeclamptic pregnancies

Endothelial cell activation or damage is believed to play a key role in preeclampsia (PE) and may underlie the hemostatic changes observed in this syndrome. The aim of this study was to evaluate a relationship between maternal and cord blood hemostatic disturbances in preeclamptic pregnancies. We measured the plasma levels of tissue plasminogen activator (tPA) antigen and of plasminogen activator inhibitor type 1 (PAI-1) antigen, both markers of hemostatic and endothelial function, and fibrin fragment D-dimer. Maternal blood from uncomplicated (n=42) and PEc pregnancies (n=44) were collected before delivery, and umbilical cord blood (UCB) immediately after delivery. In preeclamptic cases, UCB presented significantly higher tPA values and significantly lower PAI-1/tPA ratio. Preeclamptic women also presented significantly higher tPA, as well as PAI-1 values, when compared with normal pregnant women; no significant difference was found for D-dimer. In preeclamptic women, proteinuria (a marker of PE severity) correlated positively and significantly with tPA and PAI-1 antigen levels. An inverse relationship between maternal tPA antigen levels and fetal birth weigh in PE was also observed. Our data show that the hemostatic maternal disturbances observed in preeclamptic women have similarities with the UCB circulation, and that endothelial dysfunction is the most plausible underlying cause. Moreover, maternal hemostatic disturbances seem to be associated with the severity of PE. Further studies are needed to strength the values of tPA and PAI-1 as markers of severity in PE.

The triad psoriasis-obesity-adipokine profile.

Psoriasis is a chronic inflammatory skin disease, often associated with overweight/obesity. The adipose tissue is a complex organ that secretes several adipokines, involved in the regulation of some metabolic processes, such as lipid metabolism, glucose homeostasis, angiogenesis, blood pressure and inflammation. In obesity, the distribution and function of adipose tissue, and the adipokine profile are altered. The unbalanced production of pro‐ and anti‐inflammatory adipokines in obesity, contributes to the development of a chronic low‐grade inflammation state, which seems to favour worsening of psoriasis lesion and a poorer response to treatment. In this review, we will debate published data concerning the current knowledge about the triad psoriasis–obesity–adipokine profile.