Disease Markers - Volume 31, issue 2

Authors: Starlinger, Patrick | Alidzanovic, Lejla | Schauer, Dominic | Brugger, Philipp | Sommerfeldt, Silvia | Kuehrer, Irene | Schoppmann, Sebastian F. | Gnant, Michael | Brostjan, Christine

Article Type: Research Article

Abstract: Background: The analysis of angiogenesis factors in the blood of tumor patients has given diverse results on their prognostic or predictive value. Since mediators of angiogenesis are stored in platelets, their measurement in plasma is sensitive to inadvertent platelet activation during blood processing. Methods: Variants of blood withdrawal and plasma preparation were evaluated by ELISA for the detection of TSP-1, PF-4, VEGF and PD-ECGF. A total of 22 pancreatic cancer patients and 29 healthy …volunteers were evaluated. Results: Plasma preparation with the anticoagulant mix of citrate, theophylline, adenosine, dipyridamole (CTAD) and immediate blood processing at 4°C was required for reproducible measurements of TSP-1, PF-4 and VEGF. Blood collection by venflon or inadvertent hemolysis during blood withdrawal caused significantly elevated TSP-1 and PF-4 values. When optimized plasma preparation was applied, a significant increase of TSP-1 and VEGF in cancer patients was detected (P=0.006; P< 0.001). Conclusion: The reliable plasma analysis of circulating platelet-stored angiogenesis factors requires preparation with CTAD at 4°C and blood collection by butterfly needle. Suboptimal procedures of plasma preparation are commonly applied in clinical monitoring of angiogenesis parameters which may account for the differences in reported plasma values and may have masked their predictive or prognostic marker potential. Show more

Keywords: Angiogenesis factors, cancer, plasma, platelet derived endothelial cell growth factor, platelet factor 4, thrombospondin 1, vascular endothelial growth factor

DOI: 10.3233/DMA-2011-0798

Citation: Disease Markers, vol. 31, no. 2, pp. 55-65, 2011

Authors: Wiernicki, Ireneusz | Millo, Barbara | Safranow, Krzysztof | Gorecka--Szyld, Barbara | Gutowski, Piotr

Article Type: Research Article

Abstract: Background: Abdominal aortic aneurysms (AAAs) are characterized by presence of high proteolytic activity, atherosclerotic lesions, extensive transmural inflammation and the presence of variably sized and shaped intraluminal thrombus (ILT). Therefore, we evaluated a possible association between plasma matrix metalloproteinase-9 (MMP-9), homocysteine (Hcy), high-sensitivity C-reactive protein (hsCRP) levels and ILT thickness in patients with AAA. Methods: Plasma concentrations of MMP-9, Hcy and hsCRP were determined and ILT thickness was measured in …71 patients with AAA. They were divided into 2 groups according to ILT thickness: 34 patients with ILT mean thickness ⩾ 9 mm and 37 patients with ILT < 9 mm. Results: Plasma MMP-9 and CRP concentrations in patients with thin ILT were significantly higher than in group with thick ILT (medians 610 vs. 485 ng/mL, p=0.00003, and 7.7 vs. 3.3 mg/L, p < 0.00001, respectively). In contrast, plasma Hcy concentrations in patients with thin ILT were significantly lower than in the group with thick ILT (medians 14.3 vs. 19.2 μmol/L, p < 0.00001). Multiple regression models adjusted for age and AAA diameter showed that thin ILT is an independent predictor of high MMP-9 and CRP concentrations, while thick ILT predicts high Hcy concentrations. Conclusions: Association of higher plasma levels of MMP-9 and CRP with thin ILT may be related to two phenomena: thin thrombi convey more elastolysis-stimulating factors from blood to the AAA wall and thin thrombi convey more factors involved in proteolysis and inflammation from AAA wall to blood. The association of thin ILT with lower plasma Hcy concentrations may be related to the role of Hcy as a prothrombotic marker and needs further research. Show more

Keywords: Abdominal aortic aneurysm, intraluminal thrombus, plasma biomarkers

DOI: 10.3233/DMA-2011-0799

Citation: Disease Markers, vol. 31, no. 2, pp. 67-74, 2011

Authors: Martínez-Sales, Vicenta | Sánchez-Lázaro, Ignacio | Vila, Virtudes | Almenar, Luis | Contreras, Teresa | Reganon, Edelmiro

Article Type: Research Article

Abstract: Introduction and Aims: Acute and chronic heart failure may manifest different degrees of endothelial damage and angiogenesis. Circulating endothelial cells (CEC) have been identified as marker of vascular damage. The aim of our study was to evaluate the evolution of the CEC at different stages of patients with heart failure. We also investigated a potential correlation between CEC and markers of vascular damage and angiogenesis. Methods: We studied 32 heart failure patients at hospital admission (acute phase) and …at revision after 3 months (stable phase) and 32 controls. Circulating markers of endothelial damage (CEC; von Willebrand factor, vWF and soluble E-selectin, sEsel) and angiogenesis (vascular endothelial growth factor, VEGF and thrombospondin-1) were quantified. Results: Levels of CEC, vWF, sEsel and VEGF are significantly higher in heart failure patients than in controls. Levels of CEC (36.9 ± 15.3 vs. 21.5 ± 10.0 cells/ml; p< 0.001), vWF (325 ± 101 vs. 231 ± 82%; p< 0.001) and VEGF (26.3 ± 15.2 vs. 21.9 ± 11.9 ng/ml; p< 0.001) are significantly higher in the acute phase than in the stable phase of heart failure. CEC levels correlate with vWF and VEGF. Results show than 100% of patients in acute phase and 37.5% in stable phase have levels of CEC higher than the 99th percentile of the distribution of controls (16 cells/ml). Therefore, increases in CEC represent a relative risk of 9.5 for heart failure patients suffering from acute phase. Conclusions: CEC, in addition to being elevated in heart failure, correlate with vWF levels, providing further support for CEC as markers of endothelial damage. Levels of CEC are associated with the acute phase of heart failure and could be used as a marker of the worsening in heart failure. Show more

Keywords: Heart failure, circulating endothelial cells, endothelial dysfunction, angiogenesis

DOI: 10.3233/DMA-2011-0801

Citation: Disease Markers, vol. 31, no. 2, pp. 75-82, 2011

Authors: Díaz-Olguín, Lizbeth | Coral-Vázquez, Ramón Mauricio | Canto-Cetina, Thelma | Canizales-Quinteros, Samuel | Ramírez Regalado, Belem | Fernández, Genny | Canto, Patricia

Article Type: Research Article

Abstract: Preeclampsia is a specific disease of pregnancy and believed to have a genetic component. The aim of this study was to investigate if three polymorphisms in eNOS or their haplotypes are associated with preeclampsia in Maya mestizo women. A case-control study was performed where 127 preeclamptic patients and 263 controls were included. Genotyped and haplotypes for the -768T→C, intron 4 variants, Glu298Asp of eNOS were determined by PCR and real-time PCR …allelic discrimination. Logistic regression analysis with adjustment for age and body mass index (BMI) was used to test for associations between genotype and preeclampsia under recessive, codominant and dominant models. Pairwise linkage disequilibrium between single nucleotide polymorphisms was calculated by direct correlation r^{2} , and haplotype analysis was conducted. Women homozygous for the Asp298 allele showed an association of preeclampsia. In addition, analysis of the haplotype frequencies revealed that the -786C-4b-Asp298 haplotype was significantly more frequent in preeclamptic patients than in controls (0.143 vs. 0.041, respectively; OR =3.01; 95% CI = 1.74–5.23; P =2.9 × 10^{-4} ). Despite the Asp298 genotype in a recessive model associated with the presence of preeclampsia in Maya mestizo women, we believe that in this population the -786C-4b-Asp298 haplotype is a better genetic marker. Show more

Keywords: Preeclampsia, maya-mestizo women, polymorphisms of eNOS, eNOS haplotypes.

DOI: 10.3233/DMA-2011-0804

Citation: Disease Markers, vol. 31, no. 2, pp. 83-89, 2011

Authors: Paone, Gregorino | Conti, Vittoria | Vestri, Annarita | Leone, Alvaro | Puglisi, Giovanni | Benassi, Fulvio | Brunetti, Giuseppe | Schmid, Giovanni | Cammarella, Ilio | Terzano, Claudio

Article Type: Research Article

Abstract: The pivotal role of neutrophils and macrophages in smoking-related lung inflammation and COPD development is well-established. We aimed to assess whether sputum concentrations of Human Neutrophil Peptides (HNP), Neutrophil Elastase (NE), Interleukin-8 (IL-8), and Metalloproteinase-9 (MMP-9), major products of neutrophils and macrophages, could be used to trace airway inflammation and progression towards pulmonary functional impairment characteristic of COPD. Forty-two symptomatic smokers and 42 COPD patients underwent pulmonary function tests; sputum samples were …collected at enrolment, and 6 months after smoking cessation. HNP, NE, IL-8, MMP-9 levels were increased in individuals with COPD (p < 0.0001). HNP and NE concentrations were higher in patients with severe airways obstruction, as compared to patients with mild-to-moderate COPD (p =0.002). A negative correlation was observed between FEV_{1} and HNP, NE and IL-8 levels (p < 0.01), between FEV_{1} /FVC and HNP, NE and IL-8 levels (p< 0.01), and between NE enrolment levels and FEV_{1} decline after 2 years (p =0.04). ROC analysis, to discriminate symptomatic smokers and COPD patients, showed the following AUCs: for HNP 0.92; for NE 0.81; for IL-8 0.89; for MMP-9 0.81; for HNP, IL-8 and MMP-9 considered together 0.981. The data suggest that the measurement of sputum markers may have an important role in clinical practice for monitoring COPD. Show more

Keywords: Airway inflammation, Chronic obstructive pulmonary disease, neutrophil elastase, human neutrophil peptides, interleukin-8, likelihood ratio, metalloproteinase-9, sputum, receiver operating characteristic analysis, smoking

DOI: 10.3233/DMA-2011-0807

Citation: Disease Markers, vol. 31, no. 2, pp. 91-100, 2011

Authors: Chen, Yung-Che | Chin, Chien-Hung | Liu, Shih-Feng | Wu, Chao-Chien | Tsen, Chia-Cheng | Wang, Yi-Hsi | Chao, Tung-Ying | Lie, Chien-Hao | Chen, Chung-Jen | Wang, Chin-Chou | Lin, Meng-Chih

Article Type: Research Article

Abstract: Objective: To identify patients at high risk of relapse after anti-tuberculosis (TB) therapy or with poor long-term outcomes. Methods: Fifty-one patients with pulmonary TB: 7 were classified as high association with both cavitations on initial chest radiography and positive sputum smear/cultures after two months of anti-TB treatment (HA group); 19 medium association (MA, one risk alone); and 25 low association (LA, neither risk). Serum interferon (IFN)-γ-inducible protein 10 (IP-10), interleukin-17 (IL-17), and C-reactive protein …levels were investigated. Results: There was a trend towards higher serum IP-10 levels (p=0.042) for HA patients throughout the 6-month treatment period. Month-2 IP-10 levels were higher in the HA than in the MA/LA group (656.2 ± 234.4 vs. 307.6 ± 258.5 pg/ml, adjusted p =0.005). Receiver operating characteristic curves showed that the risk of relapse was well-captured by month-2 IP-10 levels at a cut-off value of 431 pg/ml (AUC=0.857, 95% CI 0.75–0.97, p =0.003). Month-2 serum IL-17 levels were lower in non-survivors than survivors (15.7 ± 2.9 pg/ml vs. 24.6 ± 8.2 pg/ml, p=0.001). Multivariate analysis demonstrated that a month-2 serum IL-17 level of ⩽ 17 pg/ml (p =0.026) was independently associated with all-cause mortality. Conclusions: Serum IP-10 and IL-17 levels after 2 months of anti-TB treatment may be biomarkers for estimating risk of both cavitation and delayed sputum conversion, and for predicting long-term mortality, respectively. Show more

Keywords: Pulmonary tuberculosis, interferon-γ-inducible protein 10, interleukin-17, caviation, mortality

DOI: 10.3233/DMA-2011-0808

Citation: Disease Markers, vol. 31, no. 2, pp. 101-110, 2011

Authors: Kosanovic, Maja M. | Goc, Sanja R. | Potpara, Goran S. | Jankovic, Miroslava M.

Article Type: Research Article

Abstract: Prostate specific antigen (PSA) exhibits pronounced heterogeneity in both primary structure and glycan composition, resulting in the existence of different molecular forms. Investigation of PSA structure is a demanding task facing limitations due to inadequate sensitivity of analytical techniques and low concentrations of the different forms. This study aimed to profile free PSA (fPSA), especially lower molecular mass species lacking detailed classification, in normal seminal plasma and in sera from subjects with benign …hyperplasia (BPH) or cancer of the prostate (PCa) as samples of known clinical relevance. fPSA forms were separated from complex proteomes on chips with immobilized anti-fPSA antibody followed by detection using surface-enhanced laser desorption/ionization time of flight mass spectrometry. At least 39 fPSA-immunoreactive species, ranging from 3–29 kDa were detected in seminal plasma. General fPSA profiles in seminal plasma and sera were similar, but differed in the abundance and presence of particular peaks/clusters of the lower molecular mass species. No striking difference in fPSA forms was observed between BPH and PCa samples, but some distinct peaks varied in intensity and frequency within or between groups. Obtained data verify fPSA heterogeneity that might be important for better exploration of all their molecular and marker potentials. Show more

Keywords: Free PSA, molecular forms, prostate cancer, benign prostatic hyperplasia, SELDI-TOF/MS

DOI: 10.3233/DMA-2011-0809

Citation: Disease Markers, vol. 31, no. 2, pp. 111-118, 2011