Evaluation of the Vitros Syphilis TPA chemiluminescence immunoassay as a first-line method for reverse syphilis screening.

We report here the results of the diagnostic performances of Vitros Syphilis TPA (a chemiluminescence treponemal assay) compared with those of two treponemal enzyme immunoassays and of traditional versus reverse syphilis algorithms. Ease of use, automation, and high throughput make the Vitros Syphilis TPA assay a good choice for syphilis screening in high-volume laboratories.

The roles of cells and cytokines in the pathogenesis of psoriasis.

Psoriasis is considered an immune chronic disease in which T cells are accepted as important. Nowadays, it is believed that psoriasis is most likely a T helper (Th)1/Th17 induced inflammatory disease. However, some other cells, such as endothelial cells, dendritic cells, monocytic cells, neutrophils, keratinocytes, and several cytokines, appear to have, at different stages of the disease, an important role in its pathogenesis. For instance, the response to psoriasis therapy is dependent not only on the inactivation of Th1 and Th17 immune responses but also on the inactivation of dendritic cell products. Moreover, interleukin (IL)-23 deregulation appears to be an independent factor in the pathogenesis of psoriasis. Indeed, currently, the IL-23/Th17 axis is believed to be crucial in psoriasis pathogenesis, and its inhibition appears to be important for therapeutic achievement. This review presents the roles and interactions of cells and cytokines that are related to psoriasis pathogenesis.

Erythroid disturbances before and after treatment of Portuguese psoriasis vulgaris patients: a cross-sectional and longitudinal study.

BACKGROUND: A few studies in psoriasis vulgaris patients have reported changes suggesting red blood cell (RBC) damage is linked to neutrophil activation, oxidative stress, and psoriasis worsening. OBJECTIVE: The aim of this study was to evaluate erythroid disturbances in Portuguese psoriasis vulgaris patients, before, during, and after treatment. METHODS: A cross-sectional study (n = 73 patients vs 40 healthy control subjects) followed by a longitudinal study (n = 47 patients) was performed, with assessments before, and at 3, 6, and 12 weeks of therapy (10 patients started topical treatment, 17 narrow-band UVB, and 20 photochemotherapy [psoralen plus UVA; PUVA]). Evaluations included hematologic data, total bilirubin levels, membrane-bound hemoglobin (MBH), membrane protein band 3 profile, total plasma antioxidant status (TAS), lipid peroxidation (thiobarbituric acid [TBA] assay), elastase, lactoferrin, and C-reactive protein (CRP). RESULTS: Before treatment, patients presented with higher leukocyte/neutrophil and reticulocyte counts, elastase, lactoferrin, TBA, TBA/TAS, reticulocyte production index, total bilirubin and MBH values, lower RBC and hematocrit, higher percentages of high-molecular-weight aggregates, and lower percentages of band 3 monomer. After treatment, we observed a reversal in most of the parameters. However, patients still presented with values suggestive of accelerated RBC damage, removal, and production, as most of the parameters were still higher than those in the control group; the same occurred with CRP. CONCLUSION: Our data suggest that psoriasis vulgaris triggers an inflammatory response, with release of acute-phase reactants, reactive oxygen species, cationic proteins, and proteases, leading to enhanced RBC damage/aging and, ultimately, to enhanced RBC removal. These assumptions were strengthened by the observation that, with treatment, all of these changes were reversed, the inflammation was reduced, the production of reticulocytes was increased, and the RBCs presented changes usually observed in younger/less damaged RBCs. These erythroid changes were enhanced with PUVA therapy, probably due to the more pronounced clearing of the lesions, as suggested by Psoriasis Area and Severity Index (PASI) scores. Finally, after treatment, a residual inflammation still persisted that might contribute to the observed erythroid disturbances.

Psoriasis therapy and cardiovascular risk factors: a 12-week follow-up study.

BACKGROUND: Psoriatic patients present with an increased frequency of cardiovascular events. OBJECTIVE: To study the impact of psoriasis duration and therapy on traditional and new cardiovascular risk factors. STUDY DESIGN: A longitudinal study performed between 2005 and the first trimester of 2008. Each patient was followed up for 12 weeks, and was observed before and 3, 6, and 12 weeks after starting therapy. SETTING: Patients attending the Dermatology Service, University Hospital of Coimbra, Coimbra, Portugal were enrolled. SUBJECTS: Thirty-four patients with psoriasis vulgaris and 37 healthy volunteers as controls. MAIN OUTCOME MEASURES: Psoriasis Area and Severity Index (PASI); lipid profile, oxidized low-density lipoprotein (oxLDL), oxLDL/low-density lipoprotein (LDL), total antioxidant status, lipid peroxidation, C-reactive protein (CRP), and circulating levels of adiponectin. INTERVENTION: Ten patients started therapy with topical treatment, 11 with narrow-band UVB radiation (NB-UVB), and 13 with psolaren plus UVA (PUVA). RESULTS: Before starting therapy, psoriatic patients presented with several risk changes in their lipid profiles, and significantly higher CRP, oxLDL, and oxLDL/LDL, and lower adiponectin levels (vs control subjects), which may further contribute to inflammation and atherogenesis. After treatment of the patients, although no significant differences were observed in the lipid profile compared with baseline, some changes suggested that the treatment could somehow alter lipid metabolism, as the reduction in high-density lipoprotein cholesterol (HDL-C) and apolipoprotein A and the increase in the atherogenic index cholesterol/HDL-C maintained an even higher significance (as shown by p-values) when compared with the control group. After topical therapy, there was a significant reduction in thiobarbituric acid reactivity only, suggesting that the reduction in the hyperproliferative process within the lesions is important for lipid peroxidation. After NB-UVB therapy, oxLDL/LDL, cholesterol/HDL-C, lipoprotein (a) [Lp(a)], and CRP remained higher than in the control subjects, reflecting persistent inflammation and atherogenic risk. After PUVA treatment, there was a significant reduction in Lp(a), associated with an almost significant increase in apolipoprotein-B (p = 0.054); these changes were not observed after NB-UVB treatment. However, after PUVA and NB-UVB treatment, CRP and, in the NB-UVB group, oxLDL/LDL were persistently higher than controls. CONCLUSION: Our data show that psoriatic patients present with several lipid profile changes that seem to be related to the severity of the disease and/or the treatment used. Mild psoriasis patients receiving topical treatment presented before starting therapy with a lipid profile similar to controls, whereas those undergoing NB-UVB and PUVA, who had higher PASI scores, presented with several risk factors. Moreover, PUVA therapy seems to interact in a different way with lipids that might result from an interaction of psoralen with plasma lipids, namely Lp(a). Inflammation, a hallmark of psoriasis, also seems to be related to psoriasis severity. Both NB-UVB and PUVA were effective, as shown by the reduction in PASI score, as well as in the oxidative and inflammatory stress markers. However, after NB-UVB and PUVA, a low-grade inflammatory process still persisted, which might be related to the duration of remission of the disease.

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.

Lipid profile in Portuguese obese children and adolescents: interaction of apolipoprotein E polymorphism with adiponectin levels.

OBJECTIVE: To evaluate how the lipid profile associates with apolipoprotein (apo) E gene polymorphism, plasma adiponectin level, and body mass index (BMI) z score in Portuguese youth. DESIGN: Transversal cohort study. SETTING: Hospital de São João and Hospital de Crianças Maria Pia, Porto, Portugal, between May 2006 and March 2007. PARTICIPANTS: One hundred thirty-eight obese children and adolescents (62 boys; mean age, 10.8 years [range, 4-16 years]). Participants were grouped according to (1) apo E polymorphism: presence of the apo E alleles 2 or 4 in E2 (n = 11) and E4 (n = 31) carriers, respectively, or as E3/E3 (n = 94) (carriers of E2/E4 [n = 2] were excluded from apo E analysis because they carry both alleles) and (2) BMI z score: group 1, BMI z score less than 2 (n = 31); group 2, BMI z score of 2 or more and less than 2.5 (n = 65); and group 3, BMI z score of 2.5 or more (n = 42). MAIN OUTCOME MEASURES: Lipid variable comparisons between apo E polymorphism and BMI z score groups and influence of BMI z score and adiponectin level, adjusted for apo E polymorphism, on total cholesterol to high-density lipoprotein cholesterol and apo A-I to apo B ratios. RESULTS: E4 carriers presented with a worse lipid profile when compared with E2 and E3/E3 carriers. There was also a clear risk of worsening for the group with the highest BMI z score. Apolipoprotein E polymorphism, BMI z score, and adiponectin level were significantly associated with total cholesterol to high-density lipoprotein cholesterol (standardized beta coefficient = 0.283, 0.354, and -0.292, respectively; P < .001 for all) and apo A-I to apo B (standardized beta coefficient = -0.372, -0.284, and 0.327, respectively; P < .001 for all) ratios. CONCLUSION: Our data suggest a more atherogenic lipid profile for some apo E genotypes and for increasing BMI z score, whereas adiponectin level seems to play a protective role.

Erythrocyte changes in preeclampsia: relationship between maternal and cord blood erythrocyte damage.

Our aim was to evaluate red blood cell (RBC) changes in normal and preeclamptic cases, and to assess the relationship between maternal and umbilical cord blood (UCB) changes. We evaluated markers of RBC damage: membrane bound hemoglobin (MBH) and band 3 profile - high molecular weight aggregates (HMWSAg), monomer and proteolytic fragments. RBCs are marked for removal by a rise in MBH and in HMWAg. Preeclamptic mothers had significantly higher MBH, RBC count, hemoglobin, hematocrit, reticulocytes and reticulocyte production index (RPI). In UCB from newborns of preeclamptic mothers, we found similar HMWAg, RBC count, hemoglobin and hematocrit; significantly higher MBH, mean cell hemoglobin concentration, mean cell volume, RPI and reticulocyte count. Maternal MBH and HMWAg values were positively and significantly correlated with MBH and HMWAg values in UCB, in normal as well as in preeclamptic pregnancies; in preeclampsia, a significant positive correlation between UCB and maternal bilirubin, and between RPI and proteinuria were found. We conclude that markers of RBC damage/production are altered in preeclampsia, in both UCB and maternal circulation. Our data show similarities between UCB and maternal RBC changes, as suggested by the correlations of markers of RBC damage.

Changes in red blood cells membrane protein composition during hemodialysis procedure.

Our aim was to evaluate the influence of the hemodialysis (HD) procedure in red blood cells (RBC) membrane protein composition. We evaluated hematological data (RBC count, hemoglobin concentration, and hematimetric indices) and RBC membrane protein composition (linear and exponential gradient polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate [SDS-PAGE] followed by densitometry analysis of RBC membrane proteins) before and immediately after the HD procedure in 20 patients (10 responders and 10 non-responders to recombinant human erythropoietin therapy [rhEPO]) and 26 healthy controls. Before HD, patients presented anaemia and significant changes in membrane protein composition, namely, a statistically significant reduction in spectrin associated with a significant increase in bands 6, as well as an altered membrane protein interaction (protein 4.1/spectrin, protein 4.1/band 3, protein 4.2/band 3 and spectrin/band 3). After HD, we found that patients showed a statistically significant increase in RBC count and hemoglobin, a further and statistically significant decrease in spectrin, an increase in band 3, and an altered spectrin/band 3 ratio. When comparing responders and non-responders patients after HD, we found that the non-responders presented a trend to a higher reduction in spectrin. Our data suggest that HD procedure seems to contribute to a reduction in spectrin, which is normally associated with a reduction in RBC deformability, being that reduction in spectrin is higher in non-responder patients.

Neutrophil activation and resistance to recombinant human erythropoietin therapy in hemodialysis patients.

AIM: The aim of this work was to evaluate the neutrophil activation state in chronic kidney disease (CKD) patients under hemodialysis, and its linkage with resistance to recombinant human erythropoietin (rhEPO) therapy. METHODS: We studied 63 CKD patients under hemodialysis and rhEPO treatment (32 responders and 31 non-responders to rhEPO therapy). In 20 of the CKD patients (10 responders and 10 non-responders to rhEPO therapy), blood samples were also collected immediately after dialysis. Twenty-six healthy volunteers were included in a control group. Hemoglobin levels, total and differential leukocyte counts, and circulating levels of C-reactive protein (CRP), elastase and lactoferrin were measured in all patients and controls. RESULTS: Compared with controls, CKD patients presented with significantly higher CRP, neutrophil and elastase levels. When we compared the 2 groups of patients, we found that non-responders presented statistically significantly higher elastase plasma levels. A positive significant correlation was found between elastase levels and weekly rhEPO dose and CRP serum levels. After the hemodialysis procedure, a statistically significant rise in elastase, lactoferrin and, elastase/neutrophil and lactoferrin/neutrophil ratios were found. CONCLUSIONS: Our data show that CKD patients under hemodialysis present higher elastase levels (particularly in non-responding patients), which could be related to the rise in neutrophils, and to be part of the enhanced inflammatory process found in these patients.

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.

Altered erythrocyte membrane protein composition in chronic kidney disease stage 5 patients under haemodialysis and recombinant human erythropoietin therapy.

Our aim was to evaluate red blood cell (RBC) membrane protein composition in chronic kidney disease (CKD) stage 5 patients under haemodialysis (HD) and recombinant human erythropoietin (rhEPO) therapy, and its linkage to rhEPO hyporesponsiveness. We evaluated in 63 CKD stage 5 patients (32 responders and 31 non-responders to rhEPO therapy) and in 26 healthy controls RBC count, haematocrit, haemoglobin concentration, haematimetric indices, reticulocyte count, reticulocyte production index, RBC osmotic fragility test and membrane protein analyses. CKD stage 5 patients presented significant changes in membrane protein composition, namely a reduction in spectrin, associated to altered protein 4.1/spectrin and spectrin/band 3 ratios. Non-responder CKD stage 5 patients were more anaemic, with more microcytic and anisocytic RBCs, than responders; significantly altered ankyrin/band 3 and spectrin/ankyrin ratios were also observed. CKD stage 5 patients under HD are associated with an altered protein membrane structure, which seems to the disease itself and/or to the interaction with HD membranes.

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.

Presence of cytosolic peroxiredoxin 2 in the erythrocyte membrane of patients with hereditary spherocytosis.

We studied 82 Portuguese individuals, 57 with hereditary spherocytosis (HS) and 25 unaffected controls. We performed standardized diagnosis tests, including electrophoretic membrane protein analysis to identify and quantify protein deficiencies underlying HS. Membrane bound hemoglobin (MBH) and band 3 profiles were determined as oxidative stress and aging markers. A protein of about 22 kDa, present in 21 of 57 HS patients, but not in controls, was identified as peroxiredoxin 2 (Prx2), by mass-spectroscopy and by immunoblotting. Human erythrocyte Prx2 is a peroxiredoxin with thiol-specific antioxidant activity. The presence of Prx2 in erythrocyte membranes was linked to higher levels of oxidative stress, as reflected by significantly increased MBH in those HS patients. No relation with HS clinical severity was observed and Prx2 was detected in all types of membrane protein abnormalities. Prx2 membrane linkage is associated with a higher oxidative stress susceptibility of HS erythrocytes.

Role of prohepcidin, inflammatory markers and iron status in resistance to rhEPO therapy in hemodialysis patients.

The aim of our study was to assess possible relations between prohepcidin, iron status and inflammatory markers in hemodialysis (HD) patients, as well as its association with resistance to recombinant human erythropoietin (rhEPO) therapy. Fifty HD patients and 25 healthy controls were enrolled in the study. Among HD patients, 25 were non-responders and 25 were responders to rhEPO therapy. Complete blood cell count, reticulocyte count, and circulating levels of ferritin, iron, transferrin saturation, C-reactive protein (CRP), soluble interleukin (IL)-2 receptor (s-IL2R), soluble transferrin receptor (s-TfR), IL-6 and prohepcidin were measured in all patients and controls. HD patients showed higher circulating levels of ferritin, s-TfR, CRP, IL-6, s-IL2R and prohepcidin, and lower levels of transferrin compared to healthy controls. Higher levels of s-TfR, CRP and lower levels prohepcidin were observed among non-responders compared to responders. Prohepcidin levels correlated negatively with s-TfR and reticulocyte count. The weekly rhEPO/kg dose was found to be positively correlated with CRP, hemoglobin and s-TfR. In conclusion, our data show that a close interaction exists between inflammation, iron status and prohepcidin serum levels that ultimately regulate intracellular iron availability. Prohepcidin and s-TfR, together with CRP, may prove to be good markers of resistance to rhEPO therapy in HD patients.

Inflammation, T-cell phenotype, and inflammatory cytokines in chronic kidney disease patients under hemodialysis and its relationship to resistance to recombinant human erythropoietin therapy.

BACKGROUND: Resistance to recombinant human erythropoietin (rhEPO) occurs in some chronic kidney disease (CKD) patients, which may be due to enhanced systemic inflammatory response and to the erythropoiesis-suppressing effect of pro-inflammatory cytokines, some of which are produced by T cells. AIM OF STUDY: The aim of this study was to investigate the relationship between resistance to rhEPO therapy in hemodialysis CKD patients and inflammatory markers [C-reactive protein (CRP), soluble interleukin (IL)-2 receptor (sIL2R), and serum albumin levels], blood cell counts, T-cell phenotype, cytokine production by T cells, and serum cytokine levels. MATERIALS AND METHODS: We studied 50 hemodialysis CKD patients, 25 responders and 25 nonresponders to rhEPO, and compared them to each other and with 25 healthy controls. When compared to controls, CKD patients showed increased serum levels of CRP, IL-6, and sIL2R and a T-cell lymphopenia, due to decreased numbers of both CD4+ and CD8+ T cells. T cells from CKD patients had an immunophenotype compatible with chronic T-cell stimulation as shown by the increased percentage of CD28-, CD57+, HLA-DR+, CD28-HLA-DR+, and CD57+ HLA-DR+ T cells and produce higher levels of IL-2, INF-gamma, and TNF-alpha after short-term in vitro stimulation, although Th1 cytokines were not detectable in serum. Statistically significant differences were found between responders and nonresponders to rhEPO therapy for total lymphocyte and CD4+ T-lymphocyte counts, albumin (lower in nonresponders) and CRP (higher in nonresponders) levels. CONCLUSION: CKD patients under hemodialysis present with raised inflammatory markers and decrease of total lymphocyte and CD4+ T-lymphocyte counts when compared with controls. Some of those markers are even further enhanced in nonresponders to rhEPO therapy patients, but resistance to this therapy cannot be justified by a Th1 polarized T-cell response.