Activated platelets in central venous catheters: a flow cytometry and numerical simulation approach

The central venous catheter that is inserted in patients undergoing hemodialysis can cause hemodynamic instability and trigger complications such as thrombus formation. The objective of this study was to investigate hemostatic and numerical influences on thrombus formation in patients undergoing hemodialysis with a central venous catheter. Participants were assigned to three groups: I: clinical and laboratorial healthy individuals matched by sex and age (controls); II: participants after one month of insertion of the catheter and III: participants after 4 months of insertion of the catheter. Platelet activation was investigated by GPIIb/IIIa and p-selectin expressions using flow cytometry. A three-dimensional model of the catheter was constructed in the numerical simulation for the calculation of partial differential equation of a platelet activation model. A significant difference was detected by the expression of p-selectin comparing the group I (33.42 ± 4.74), group II (40.79 ± 5.54) and group III(51.00 ± 7.21) (p < 0.0001). The median values for GPIIb/IIIa were 10426 (10029-10721), 13921 (13412-15652) and 19946 (18714-21815) after catheter insertion (p < 0.0001), for groups I, II and III, respectively. Excluding the first arterial orifice, venous orifices tend to have greater platelet activation when compared to the other arterial orifices. The results of this study showed the influence of arterial and venous lateral orifices in stimulating the development of thrombi associated with the activation of platelet markers the longer the catheter was used.

Intercellular mediators in bone remodeling regulation in the experimental renal pathology / Mediadores intercelulares de la regulación de la remodelación ósea en un modelo experimental de patología renal

Bone metabolism disorders are characterized by an imbalance of bone resorption and formation in the bone remodeling process. Glucocorticoids that are used to treat kidney diseases exacerbate these disorders. P-selectin and galectin-3 are molecules involved in the sclerotic process in kidney, whereas bone resorption is regulated by the interaction between the nuclear factor activator kappa b receptor (RANK), its ligand (RANKL) and the RANKL decoy receptor osteoprotegerin (OPG). The aim of this study was to investigate the cellular and molecular mechanisms of disruption of bone remodeling regulation processes, reflected by intercellular mediators (RANKL, OPG, P-selectin and galectin-3) in chronic kidney disease experimental model treated with glucocorticoids. Rats were divided into four groups of 10 animals each. The first group, the control group, included intact animals. The second group consisted of rats with impaired bone remodeling resulting from chronic kidney disease (experimental group (CKD). The third group was a group of animals with impaired bone remodeling due to exposure to glucocorticoids (experimental group (GCs)). The fourth group consisted of rats with impaired bone remodeling in chronic kidney disease, followed by exposure to glucocorticoids (experimental group (CKD + GCs)). The effects of CKD and glucocorticoid were evaluated biochemically, histologically and by measuring bone density. An enzymelinked immunoassay was used to measure intercellular mediator levels in the serum. The bone density in the experimental groups was reduced compared to the control group. RANKL levels in animals of three experimental groups were higher than in intact animals. Serum levels of OPG were higher in CKD and GCs groups than in intact animals. At the same time, in the animals' blood serum of the CKD + GCs group, the levels of OPG were lower, than those in animals from the control group. The levels of galectin-3 in the serum of the experimental groups GCs and CKD + GCs were lower than in intact animals. The serum levels of galectin-3 in animals of the CKD group were higher than those in animals from the control group. The levels of P-selectin were lower in the serum of the GCs group than in intact animals. At the same time, the levels of P-selectin were higher in the CKD and CKD + GCs groups, than those in animals from the control group. In conclusion, the study of the complex system of bone remodeling regulation, which includes many factors and their interactions, may lead to the development of new methods for treating patients with chronic kidney disease in order to prevent osteoporosis in the future. (AU)

Las enfermedades metabólicas óseas se caracterizan por un desequilibrio en el proceso de remodelación ósea en los que participan mediadores tales como receptor del activador del factor nuclear- kappa- b (RANK), su ligando (RANKL) y la osteoprotegerina (OPG). Los glucocorticoides, recuentemente empleados en el tratamiento de la enfermedad renal crónica, exacerban este desequilibrio. En la enfermedad esclerótica renal, las moléculas de adhesión celular P-selectina and galectina-3 tienen un rol fundamental. El objetivo de esta trabajo fue estudiar las alteraciones en los mediadores de la remodelación ósea (RANKL, OPG, P-selectina and galectina-3) en un modelo de enfermedad renal crónica con tratamiento glucocorticoideo. Ratas Wistar hembras fueron divididos en 4 grupos: control (C); enfermedad renal crónica con afección de la remodelación ósea (ERC); animales con afección de la remodelación ósea expuestos a glucocorticoides (GC); enfermedad renal crónica con afección de la remodelación ósea tratados con glucocorticoides (ERC+GC). Los efectos de la ERC y los GC fueron evaluados bioquímicamente, histológicamente y por medición de la densidad ósea. RANKL, OPG, Pselectina and galectina-3 se cuantificaron en muestras de sangre venosa empleando enzimoinmuno análisis. En los 3 grupos experimentales la densidad ósea se evidenció reducida y los niveles séricos de RANKL elevados respecto al grupo control. Los niveles de OPG en los grupos ERC y GC fueron superiores mientras que en el grupo ERC+GC menores respecto a los animales controles. Galectina 3 plasmática en GC y ERC+GC se encontró reducida y aumentada en los animales ERC, en comparación con los animales controles. La concentración sérica de P-selectina sérica fue mayor en los grupos ERC y ERC+GC, y menor en los animales GC respecto a los niveles plasmáticos de los animales intactos. El avance del conocimiento sobre la regulación de la remodelación ósea a través de la interacción de mediadores sistémicos, en un futuro, puede conducir al desarrollo de nuevas estrategias terapéuticas para la prevención de la osteoporosis en pacientes con enfermedad renal crónica. (AU)

Correlations of inhaled NO with the cTnI levels and the plasma clotting factor in rabbits with acute massive pulmonary embolism

Abstract Purpose: To investigate the correlation of inhaled nitric oxide (NO) on plasma levels of cardiac troponin I (cTnI) and von Willebrand factor (vWF), glycoprotein (GP) IIb/IIIa, granule membrane protein 140 (GMP-140) in rabbits with acute massive pulmonary embolism (PE). Methods: Thirty apanese white rabbits were divided into 3 groups, thrombus were injected in model group (n = 10), NO were inhalated for 24 h after massive PE in NO group (n = 10), saline were injected in control group (n = 10). The concentrations of vWF, GP IIb/IIIa, GMP-140 and cTnI were tested at 4, 8, 12, 16, 20, and 24 h, Correlation analyses were conducted between cTnI and vWF, GP IIb/IIIa, and GMP-140 by Pearson's correlation. Results: The concentration of cTnI and vWF, GP IIb/IIIa, and GMP-140 was increased in the model group, compared to control group. In the inhaled group, the concentrations of cTnI, vWF, GP IIb/IIIa, and GMP-140 were reduced compared to model group. There was a positive correlation between cTnI and vWF, GP IIb/IIIa, and GMP-140. Conclusion: Inhaled nitric oxide can lead to a decrease in levels of cardiac troponin I, von Willebrand factor, glycoprotein, and granule membrane protein 140, after an established myocardial damage, provoked by acute massive pulmonary embolism.

oxidative stress and platelet activation in patients infected with T. gondii

Toxoplasma gondiiis a highly frequent obligate intracellular protozoan parasite; it can cause serious problems to the public health especially pregnant females, however, the pathogenesis of this condition is not clear. Was to evaluate the status and the inter-relationship of the oxidative stress and platelet activation in patients infected with T.gondii. Thirty patients infected with T.gondii [10 acute and 20 chronic cases] and 10 healthy subjects [control group] were included in this study. Serum levels of malondialdehyde [MDA], total glutathione [t GSH], reduced glutathione [GSH], oxidized glutathione [GSSG], redox potential [RP] and soluble P-selectin [sP-selectin] were measured. EDITA blood samples were used for complete blood picture with special emphasis on platelet count and mean platelet volume [MPV]. The mean platelet volume [MPV] in patients was significantly higher than the control group, however, platelet count showed no significant difference. The serum mean values of MDA, GSSG, RP and sP-selectin in patients were significantly higher than the control subjects. On the other hand, the levels of IGSH and GSHin patients were significantly lower than in controls. MPV and platelet count showed significant positive correlations with sP-selectin concentration. Although toxoplasmosis is mostly asymptomatic, the findings of the present study strongly indicate that the occurrence of oxidative stress could be a potential mechanism of subclinical inflammatory pathology and tissue damage in these cases

Role of soluble P-selectin among type 2 diabetic patients with and without coronary heart disease

Type 2 diabetes mellitus [T2DM] is associated with a marked increase in the risk of coronary heart disease. Platelets play a significant role in coronary artery disease [CAD]. P-selectin is a component of the platelet granule membrane that is expressed on the platelet surface membrane and shed into the plasma as sP-selection on platelet activation. The current study was performed to investigate the association of soluble P-selection with inflammatory marker high sensitivity C-reactive protein [hsCRP], lipid profile and glycemic control among type 2 diabetic patients with and without coronary heart disease. This study included 31 diabetic patients with coronary artery disease [group I], 29 healthy subjects as control group [group II], and 28 diabetic patients without coronary artery disease [group III]. The age and sex of diabetic groups were matched with normal control group. Glucose, HbAlc, lipid profile, hsCRP and sP-selectin were measured. The levels of glucose, HbAlc, total cholesterol, triacylglycerol. LDL-c, hsCRP and sP-selectin level were significantly higher in the diabetic group with coronary artery disease than in the control group and the diabetic group without coronary artery disease. The level of HDL-c was significantly lower in the diabetic group with coronary artery disease than in the control group and the diabetic group without coronary artery disease. There was a significant positive correlation between level of sP-selectin and duration of CAD diabetic patients with coronary artery disease. Also there was a significant positive correlation between sP-selectin and duration of diabetes mellitus in both diabetic groups with and without coronary artery disease. ROC curve analysis for hsCRP and sP-selectin indicated that, sP-selectin had higher sensitivity and specificity than hsCRP in diabetic patient with coronary artery disease. In conclusion, measurement of soluble P-selectin seems more helpful marker of impending coronary artery insult in diabetic patients and had higher diagnostic value than hsCRP in diabetic patient with coronary artery disease

prognostic significance of galectin-3 and p-selectin in transitional cell carcinoma of the urinary bladder

Bladder carcinoma is one of the most common malignancies in urology. The most common type of the bladder cancer is transitional cell carcinoma [TCC]. TCC of bladder has a recurrence rate of more than 50%. Therefore, it is important to find some indicators that can predict for recurrence or the development of metastasis. This study investigates the prognostic significance of preoperative serum levels of galectin-3 and P-selectin in patients with transitional cell carcinoma [TCC] of the urinary bladder. Preoperative serum levels of galectin-3 and P-selectin were measured by ELISA [enzyme-linked immune-sorbent assay]. The study showed that Galectin-3 and P-selectin serum levels were higher in patients with bladder cancer than in healthy controls. Patients with metastasis had significantly higher levels of both serum Galectin-3 and P-selectin than with localized diseases. High levels of serum Galectin-3 and P-selectin were significantly associated with the clinical tumour stage. However, no significant difference detected between serum levels of galectin-3 and P-selectin in grade II and grade III TCC subgroups. Also, there is a positive correlation between serum Galectin-3 and P-selectin in both control and patient groups. Univariate analysis showed that preoperative serum level of Galectin-3, P-selectin and clinical stages were indicative for clinical progression and tumour specific survival. In a multivariate analysis, clinical stages were an independent prognostic marker for clinical progression and tumour specific survival. These results indicate that both serum Galectin-3, P-selectin levels and clinical tumour stages are closely related to poor prognosis in bladder cancer. So, their levels in bladder cancer patients may be useful in identifying patients at high risk of tumor recurrence

Study of prognostic values of soluble selectins E and P in successfully resuscitated children after cardiopulmonary arrest

Successful resuscitation of children with cardiopulmonary arrest [CPA] is often complicated by postresuscitation state related to ischemiaireperfusion injury. Studies demonstrated increase in release of soluble intercellular adhesion molecules including soluble E- and soluble P-selectins [sE- and sPselectins] during and after cardiopulmonary resuscitation [CPR]. Therefore, this study was carried out to analyze the relationship between plasma levels of sE- and sP-selectins in successfully resuscitated children after CPA and their outcome. Plasma levels of sE-and sP-selectin were measured 24 hrs after successful resuscitation of 65 children with CPA. Forty five [69.23%] patients out of 65 developed SIRS in the second day after CPR. Patients with SIRS had non significantly higher sE-selectin levels [95.58 +/- 32.38 ng/ml] than patients without SIRS [88.17 +/- 28.25 ng/ml] [p =0.67]. The mean value of sE-selectin was not significantly higher in patients with sepsis [96.53 +/- 30.62 ng/ml] than in patients without sepsis [89.39 +/- 27.57 ng/ml] [p = 0.48]. The mean value of sE-selectin was significantly higher in non survivors [147.65 +/- 43.64 ng/ml] than in survivors [87.63 +/- 25.89 ng/ml] [p <0.01]. Patients with SIRS had significantly higher sP-selectin levels [187. 12 +/- 64. 75 ng/ml] than patients without SIRS [100.42 +/- 33.70 ng/ml] [p =0.005]. The mean value of sP-selectin was significantly higher in patients with sepsis [172.40 +/- 54.43 ng/ml] than in patients without sepsis [105.60 +/- 34.35 ng/ml] [p =0.015]. The mean value of sP-selectin was not significantly higher in survivors [217.75 +/- 72.08 ng/ml] than in non survivors [175.69 +/- 51.43 ng/ml] [p = 0.175]. The total non survival was 57 [87.69%] patients. Cerebral performance at hospital discharge was good in 3 out-of 8 patients [37.5%] and unfavorable in 5 [62.5%] patients. sE-selectin had higher sensitivity [94.87%] and specificity [85.33%] in the prediction of non survival with an area under the ROC curve [95% CI] of 0.95 [0.84-0.99] and a cut-off value of 136 ng/ml. sP-selectin had higher sensitivity [87.50%] and specificity [96.55%] in the prediction of sepsis with an area under the ROC curve [95% CI] of 0.96 [0.86-0.99] and a cut-off value of 159 ng/ml. Successful CPR after cardiac arrest is associated with increase in sE- and sP-selectin and high incidence of SIRS. Estimation of sE-selectin may help detection of patients with high risk of poor outcome while estimation of sP-selectin may predict those with high risk for sepsis

Endothelial and platelet activation markers in early and convalescent phases of ischemic stroke: correlation with clinical and functional disability

The soluble form of selectins in the blood may play an important role in the pathophysiology of atherosclerotic ischemic stroke. To determine whether blood concentrations of the soluble form of selectins are elevated among patients with atherosclerotic ischemic stroke, whether their concentrations in blood correlate with clinical and functional disability and to estimate differences in their levels between lacunar and territorial strokes. We measured the serum levels of soluble E-selectin [sE-selectin] and soluble P-selectin [sP-selectin] during the early and convalescent phases of 37 patients with ischemic stroke compared to controls. We, also did correlation analysis between their levels at baseline and after 3 months with clinical and functional disability scores [National Institutes of Health Stroke Scale-NIHSS-and Barthel Index-BI-respectively]. Levels of sE-selectin and sP-selectin in stroke patients were significantly elevated compared with controls during the early phase, with significant fall in their levels below baseline measurements and below those in controls after three months. sE-selectin levels after 3 months correlated with a better functional status as measured by BI, while sP-selectin levels didn't show any correlation with clinical or functional scores. No significant differences were found in the course of sE-selectin, sP-selectin levels between lacunar and territorial strokes. The evaluation of endothelial and platelet markers would represent the pathophysiological status of stroke. This may offer the possibility of researching the application of antagonists and/ or activity modulators of some of them in ischemic brain disease therapy

relation between oxidative stress and adhesion molecules in Egyptian children and adolescents with type 1 diabetes mellitus

Antioxidant potential decreases while plasma lipid peroxidation products increase in type] diabetes mellitus. The vascular-endothelium is a major target of oxidative stress [OS]. Reactive oxygen species signal events leading to impairment of endothelial function and promotion of leukocyte adhesion to the vascular endothelium. To explore the relation between OS and adhesion molecules in type] diabetes and correlate it with the state of metabolic control, disease duration and microvascular complications [MVCs]. Thirty-eight type] diabetics were included: 22 patients with disease duration less than 5 years and 16 patients with duration of 5 years or more. Thirty healthy age and sex matched subjects served as controls. They were assessed clinically. Laboratory investigations included, random blood sugar [RBS], glycated hemoglobin [HbA[1]c], fasting lipid profile and measurement of serum malondialdehyde [MDA] as a marker of lipid peroxidation and serum soluble P-selectin as a marker of endothelial/platelet activation. Serum MDA and P-selectin were significantly elevated in type 1 diabetics compared to controls with the highest level in diabetics with disease duration of 5 years or more [p<0.0001]. Both MDA and P-selectin levels were significantly elevated in complicated compared to non complicated diabetics [P<0. 000] with strong relation to complication severity, Serum MDA level was positively correlated with serum P-selectin level in diabetics [p<0.000]. Serum MDA and P-selectin were positively and significantly correlated with disease duration [p<0. 0001], RBS [p<0. 0001, p-0. 00 respectively], HbA1c [p<0.000], diastolic blood pressure [p=0. 03, p-0.005 respectively], total cholesterol [p=0. 04, p-0. 02, respectively], triglycerides [p=0. 006, p<0.0001 respectively] and low density lipoproteins [p=0. 03, p=0.05 respectively] but negatively correlated with high density lipoproteins [p=0. 03]. On multiple regression analysis, HbA1c had the strongest effect on both MDA and P-selectin levels [P<0. 0001]. Cut off values for serum MDA and P-selectin equal to 8.035 nmoles/ml and 45. 15ng/dl respectively for early detection of diabetic MVCs were defined. Levels of M4D and P-selectin are elevated in type] diabetics with evident relation to disease duration, metabolic control and severity of MVCs. Hence both of them might act as good markers to identify diabetics who are more susceptible to develop vascular disease

Rosuvastatin and vascular dysfunction markers in pulmonary arterial hypertension: a placebo-controlled study

We investigated whether chronic rosuvastatin administration could improve the abnormalities of the circulating levels of vascular dysfunction markers in pulmonary arterial hypertension (PAH). Sixty patients, aged 13 to 60 years, with idiopathic (N = 14) or congenital heart disease-associated PAH (N = 46) were equally but randomly assigned to rosuvastatin treatment (10 mg a day, orally) or placebo for 6 months in a blind fashion. Plasma levels of P-selectin, tissue-plasminogen activator and its inhibitor as well as von Willebrand factor antigen were measured by enzyme-linked immunoassay before and after 1, 3, and 6 months of treatment. Baseline levels of biomarkers were elevated (68, 16, 45 and 46 percent increase relative to controls, for P-selectin, von Willebrand factor antigen, tissue-plasminogen activator and its inhibitor, respectively; P < 0.001). P-selectin values at baseline, 1, 3, and 6 months were 39.9 ± 18.5, 37.6 ± 14.6, 34.8 ± 14.6, and 35.4 ± 13.9 ng/mL, respectively, for the rosuvastatin group and 45.7 ± 26.8, 48.0 ± 26.9, 48.1 ± 25.7, and 45.7 ± 25.6 ng/mL for the placebo group. The P-selectin level was lower in the rosuvastatin group compared with placebo throughout treatment (P = 0.037, general linear model). A trend was observed towards a decrease in tissue-plasminogen activator in the statin group (16 percent reduction, P = 0.094), with no significant changes in the other markers. Since P-selectin is crucial in inflammation and thrombosis, its reduction by rosuvastatin is potentially relevant in the pathophysiological scenario of PAH.

Effect of Interrupted Agitation and Removal of Leukocyte on Platelet Quality during the Storage of Platelet Concentrates / 대한진단검사의학회지

BACKGROUND: This study aimed to analyze the influence of the interruption of agitation and removal of leukocytes on platelet concentrates (PCs), and determine the maximum amount of time the agitation could be interrupted without impairing PCs' effectiveness during the storage period. METHODS: Four ABO-identical random donor platelets agitated for 24 hr were pooled, and divided into 4 units, and 2 units of them were leukoreduced. Then 52 pooled units were categorized into 4 groups, non-leukoreduced continuous agitation (Non-LRCA), non-leukoreduced interrupted agitation (Non-LRIA), leukoreduced continuous agitation (LRCA), and leukoreduced interrupted agitation (LRIA), and preserved for 6 days (total 7 days). Mean platelet volume (MPV), pH, HCO3-, pO2, pCO2, CD62P, CD61, glucose, lactate, ammonia and free fatty acid were measured during the period. RESULTS: Starting from the Day 4, the pH and HCO3- of Non-LRIA group begun to decrease while the amount of lactate production, glucose consumption, and MPV increased compared to the Non- LRCA group (P<0.01). An increase in pO2 level was observed in the interrupted agitation groups as the storage period prolonged (P<0.01). The pH levels of all the units in the agitation groups remained higher than 6.4 up to Day 7, while those of the non-leukoreduction group did so only up to Day 2, but those of leukoreduction in the interrupted agitation groups did so up to Day 4. CONCLUSIONS: The interruption of agitation reduced the platelet's capacity to utilize oxygen, increasing lactate amount and reducing pH level. However, the in vitro parameters of the Non-LRIA and Non-LRCA groups on Day 2 were similar to each other and the pH level remained at 6.4 or higher, making one day of agitation interruption possible after 24 hr of agitation. With leukocytes removed, the effective agitation interruption period may become longer.

Participação das plaquetas no processo de fibrose dos pacientes com esquistossomose mansônica / Participation of platelets in the process of fibrosis in patients with mansonic schistosomiasis

O objetivo deste estudo foi avaliar a ativação plaquetária através da P-selectina e o conteúdo de PDGF-AB e TGFbeta1, nos pacientes com esquistossomose que desenvolveram fibrose (F3), naqueles que não tiveram esta manifestação (F0) e nos controles (C). Os resultados mostraram que a percentagem de P-selectina nas plaquetas sem estímulo de agonistas foi de 10,6 por cento nos F3; 11,1 por cento nos FO, e 6,3 por cento nos C e após a adição de ADP/adrenalina, foi de 44 por cento; 25,3 por cento e 42 por cento, respectivamente. A dosagem do PDGF-AB e TGFbeta1 por plaquetas foi de 11,016ng/dL (F3); 3,172 ng/dL (F0) e 5,01ng/dL (C) e, (0,012ng/dL (F3); 5,27ng/dL (F0) e 4,66ng/dL (C), respectivamente. Em relação à P-selectina, concluímos que as plaquetas dos pacientes com esquistossomoses, apesar de estarem pré ativadas, mantiveram-se responsivas aos agonistas. O TFGbeta1 não apresentou diferença entre os três grupos, enquanto o PDGF-AB foi significantemente maior no grupo F3, sugerindo a participação deste no desenvolvimento da fibrose.

The aim of this study was to evaluate platelet activation through P-selectin, and PDGF-AB and TGFbeta1 content, in schistosomiasis patients who developed fibrosis (F3) and who did not present this (F0), and in a control group (C). The results showed that the percentage of P-selectin in platelets without agonist stimulation was 10.6 percent in F3, 11.1 percent in F0 and 6.3 percent in C. After the addition of ADP/adrenaline, the percentages were 44 percent, 25.3 percent and 42 percent, respectively. The PDGF-AB and TGFbeta1 contents per platelet were 11,016ng/dl (F3), 3,172ng/dl (F0) and 5.01ng/dl (C) and 0,012ng/dl (F3), 5.27ng/dl (F0) and 4.66ng/dl (C), respectively. Concerning the P-selectin, we can conclude that platelets from patients with schistosomiasis continued to be responsive to agonists, despite being pre-activated. There were no differences in TGFbeta1 between the groups, but the PDGF-AB content was significantly higher in F3. This suggests that PDGF-AB may have some participation in the development of fibrosis.

Platelet glycoprotein [GPIIIA] gene polymorphism and aspirin resistance in patients with cerebrovascular accidents

Several large clinical trials have found that 30% to 40% of patients who had stroke were taking ASA at the time of their event. Several theories have been studied which offer possible explanation for aspirin resistance. A common T/C polymorphism at position 1565 in exon 2 of the GPIIIa gene has been implicated in increased prevalence of thrombosis and decreased responsiveness to aspirin. The aim of this study was to determine GPIIIa genotype and the degree of platelet activation in Egyptian patients with clinical aspirin resistance as evidenced by occurrence of cerebrovascular stroke in spite of taking aspirin for primary or secondary prevention using soluble P-selectin as a measure for the degree of platelet activation. Twenty nine patients with cerebrovascular stroke were enrolled from Kasr Eleini Hospital between Feb. 2006 and Jan. 2007. The enrollment criteria included documented ischemic stroke by radiological evidence [CT or MRI brain] in spite of regular aspirin use for the last month [75-325mg]. Twenty nine apparently healthy volunteers who are age and sex matched served as control. They were taking no medications. All patients and the control were subjected to clinical examination and routine labs for the estimation of blood sugar, lipid profile in addition to P-selectin level and determination of platelet glycoprotein IIIa T/C gene polymorphism. A significant difference was found in the frequency of GPIIIa gene polymorphism between patients and control; PL[A1A2] present in [55.1%] of patients and [17.2%] of control with odds ratio 5.9 CI95% 1.7-19.8 There was only one patient with PL[A1A2] genotype. The prevalence of PL[A1A2] in females was significantly higher 90% [9/10] than males 36.8% [7/19] [p 0.007] Odds ratio 15.4 [CI 95%1.13-53.62]. P-selectin levels were not significantly different between patients and control when done on the groups as a whole; however, when the groups were compared according to their platelet glycoprotein Ilia gene T/C polymorphism. Significant difference was found between groups, higher levels of P selectin was associated with GPIIIa gene polymorphism. P-selectin level was significantly higher in the heterozygous PL[A1]/ PL[A2] group of patients than homozygous PL[A1]/ PL[A2] group [p 0.000]. P-selectin was significantly lower in homozygous PL[A1]/ PL[A1] patients than in homozygous PL[A1]/ PL[A2] control [p 0.000] which may indicate better and partial, though not enough, response to aspirin. These findings indicate that patients who carry PLA2allele are less aspirin sensitive. In conclusion, aspirin prophylaxis should be individualized, taking the genotype, sex and cholesterol level in consideration. Further larger and prospective studies are needed to decide the optimal dose in variable conditions and the best test/s for follow-up of the effect of the drug

Ativação plaquetária em formas clínicas distintas da doença arterial coronariana (papel da P-selectina e de outros marcadores nas anginas estável e instável) / Platelet activation in different clinical forms of the coronary artery disease (roll of P-selectin and others platelet markers in the stable and unstable angina) / Platelet activation in different clinical forms of the coronary artery disease (roll of P-selectin and others platelet markers in the stable and unstable angina)

OBJETIVO: Os marcadores da ativação plaquetária em geral se apresentam elevados na doença arterial coronariana. Desse modo, procuramos identificar a presença e as potenciais associações de diferentes marcadores da ativação plaquetária. MÉTODOS: Estudamos pacientes com angina instável (n=28), pacientes com angina estável (n=36) e pacientes sem doença arterial coronariana (n=30); sexo e idade foram estratificados. Os níveis sangüíneos da molécula de adesão P-selectina, do thromboxane B2 e de serotonina foram medidos por imunoensaios enzimáticos. RESULTADOS: Quando comparamos os grupos, os resultados foram: a P-selectina, o thromboxane B2 e os níveis do serotonina apresentaram-se significativamente mais elevados nos pacientes com angina instável do que nos pacientes com angina estável. CONCLUSÃO: Estes marcadores da ativação plaquetária podem, portanto, identificar formas instáveis de doença arterial coronariana.

OBJECTIVE: Markers of platelet activation are elevated in coronary artery disease. We sought to identify the presence and the potential associations of different markers of platelet activation. METHODS: We studied patients with unstable angina (n=28), patients with stable angina (n=36) and patients without coronary artery disease (n=30); sex and age matched. Blood levels of the adhesion molecule P-selectin, Thromboxane B2 and Serotonin were measured by enzyme immunoassays. RESULTS: When we compared the groups the results were: sP-selectin, thromboxane B2 and serotonin levels were significantly higher in patients with unstable angina than in patients with stable angina. CONCLUSION: These markers of platelet activation were able to identify unstable forms of coronary artery disease.

Biomarcadores coronarios y evolución clínica alejada en pacientes con síndromes coronarios agudos sin elevación del segmento ST / Coronary biomarkers and long-term clinical outcome in acute coronary syndrome without ST segment elevation

Background: The use of new biomarkers improved risk stratification for patients with acute coronary syndromes (ACS). Aim: To evaluate the relationship between multiple biomarkers and long-term clinical outcome in ACS without ST segment elevation. Patients and Methods: Consecutive patients presenting with suspected ACS were studied. On admission to the emergency room, serum was obtained to determine highly sensitive C reactive protein (hsCRP), erythrocyte sedimentation rate (ESR), lipoprotein (a) (LPa) and soluble P selectin (sPS). Clinical endpoints were mortality and a composite endpoint of major adverse cardiovascular events (MACE) including death, re-infarction, and angina. Results: Seventy patients, aged 63±13 years, 54 males, were studied. Final diagnosis was unstable angina in 71% and non-ST-segment elevation myocardial infarction in 29%. MACE and mortality rate were 17% and 5.8%, respectively. We found higher plasma levels of hsCRP, ESR and Lp(a) in patients with MACE (p=0.032, p=0.015 and p=0.010, respectively). Plasma levels of hsCRP and ESR were also higher in patients who died during the follow up (p=0.002 y p=0.045, respectively). Conclusion: Plasma levels of inflammatory markers and atherosclerosis biomarkers are associated with a worse long-term clinical outcome in ACS without ST segment elevation. The inclusion of these biomarkers in the routine blood test on admission, could improve risk stratification of patients with ACS in the future.

Increased Serum Level of P-Selectin in Patients with Lichen Planus

Lichen planus (LP) is a common, pruritic and inflammatory disease of the skin, hair follicles and mucous membranes. Immunologic mechanisms, especially cell-mediated immunity, play a major role in triggering the clinical expression of the disease. P-selectin is an adhesion molecule present within endothelial cells and mediates endothelial-leukocyte interactions. Therefore, it is considered that P-selectin plays an important role in LP. The aim of our study is to research the relation between P-selectin and LP. Serum P-selectin levels were determined with the enzyme- linked immunosorbent sandwich assay method in sera from 40 LP patients and 40 healthy controls. The serum levels of P-selectin were statistically significantly higher in the patients than in healthy controls (p < 0.05), in female patients (39.32 +/- 11.34 pg/ml) than in male patients (31.93 +/- 9.83 pg/ml) (p < 0.01), and in the patients with eruptive form (40.27 +/- 9.32 pg/ml) than in those with the localised (32.83 +/- 9.93 pg/ml) and hypertrophic (31.72 +/- 8.39 pg/ml) forms (both p < 0.01). In conclusion, we found a meaningful relation between LP and serum P-selectin levels.

Effects of Cilostazol on Platelet Activation in Coronary Stenting Patients Who Already Treated with Aspirin and Clopidogrel

BACKGROUND: A recent study has shown that triple anti-platelet therapy (cilostazol+clopidogrel+aspirin) resulted in a significantly lower restenosis rate after coronary stenting than did conventional therapy (clopidogrel+aspirin). However, the anti-platelet effects of cilostazol, when combined with clopidogrel and aspirin, have not been evaluated. METHODS: Low dose cilostazol (50 mg/BID) was given to 47 patients who had already been taking clopidogrel (75 mg/day) and aspirin (100 mg/day) for more than 1 month subsequent to coronary stenting due to AMI and unstable angina. Markers of platelet activation, P-selectin and activated GPIIb/IIIa on platelets, were measured at baseline and 2 weeks after cilostazol treatment. We empirically divided patients into tertiles (low, n=16; moderate, n=14; high group, n=17), according to the baseline P-selectin expression. We then performed a comparative assessment of the anti-platelet effects of cilostazol at baseline and after 2 weeks of cilosatzol administration. RESULTS: P-selectin was significantly decreased after 2 weeks of cilostazol treatment in total patients (n=47, 3.2 +/- 2.4% to 2.0 +/- 1.9%, p=0.03). This inhibition of P-selectin expression was mainly achieved in the moderate and high P-selectin groups (low group; 1.4 +/- 0.5 to 1.9 +/- 1.3%, p> 0.05, moderate group; 2.5 +/- 0.3 to 1.3 +/- 0.3%, p 0.05). Underlying disease, cardiovascular risk factors, concomitant medication including statin, and hsCRP were not related to the degree of P-selectin expression. CONCLUSION: Our data demonstrated that cilostazol treatment in addition to conventional anti-platelet therapy provides more effective suppression of platelet P-selectin expression in patients with relatively high platelet activity.

Proteína C reactiva y trombomodulina soluble: predictores bioquímicos de mantención de ritmo sinusal a 1 año en pacientes con fibrilación auricular no valvular / C reactive protein and soluble trobomodulin: biochemical markers in the maintenance of sinusal rhythm at 1 year follow-up in patients with non valvular atrial fibrillation

Introducción: Recientemente, hemos demostrado la existencia de un estado proinflamatorio sistémico (elevación de niveles de proteína C-reactiva [PCR]) en pacientes con fibrilación auricular (FA) no valvular. Este estado inflamatorio se asocia a la pérdida de ritmo sinusal a largo plazo en estos pacientes. Hipótesis: Determinar si los niveles de PCR, como marcadores de inflamación sistémica, trombomodulina soluble (sTM,) como marcador de función endotelial, y P-selectina, como índice de activación plaquetaria, se encontraban o no, asociados a la mantención de ritmo sinusal a 1 año en pacientes con FA no valvular. Métodos: Se estudiaron 130 pacientes en forma prospectiva con FA no valvular ingresados a un hospital universitario. Ningún paciente tenía antecedente de tratamiento anticoagulante previo al ingreso. En todos ellos se midieron niveles de PCR, sTM, y marcadores hemostáticos (complejos trombina-antitrombina [TAT] y P-selectina). En todos los pacientes se realizó un ecocardiograma transtorácico y transesofágico durante las primeras 24 hrs del ingreso. Resultados: La edad promedio del grupo fue de 67±13 años (70 hombres; 46 por ciento >70 años, 59 por ciento hipertensos, 15 por ciento diabéticos y 34 por ciento con cardiopatía previa). Los niveles de PCR basales del grupo con FA fueron significativamente más altos que los niveles del grupo control de 20 sujetos (pareados por edad y sexo) en ritmo sinusal (10,5±2,2 versus 3,25 mg/L, p=0,001). Los niveles de sTM en los pacientes con FA fueron también más elevados que los controles, sin embargo, no alcanzaron significación estadística (52,2±111 versus 44±13ng/mL respectivamente). Tanto los niveles de P-selectina (219±141 ng/mL) como TAT (54±237 ng/mL) fueron significativamente más elevados que los del grupo control (p<0,05 para ambos versus el grupo control). Sin embargo, en el análisis multivariado se demostró que sólo los niveles elevados de PCR (OR =4,8 p=0,02) y sTM (OR=1,05, p=0,04), fueron predictores de mantención de ritmo sinusal a 1 año en pacientes con FA no valvular (ajustados por edad, parámetros ecocardiográficos de mantención de ritmo sinusal a largo plazo y de los niveles de TAT y P-selectina). Ni los niveles de P-selectina ni los de TAT demostraron su asociación a la mantención de ritmo sinusal a largo plazo. Conclusiones: Este estudio demuestra que los niveles de PCR y sTM contribuyen a predecir el ritmo cardíaco a largo plazo en pacientes con FA no valvular.

Evaluation of some haemostatic markers in normal pregnancies and pregnancies complicated by preeclampsia or eclampsia

To establish the plasma evolution of P-selectin, plateletfactor4 [Pf4], prothrombin fragments 1+2 [F1+2], thrombin antithrombin complex [TAT], von Willebrand factor [vWF] and blood platelet count during normal pregnancy, preeclampsia and eclampsia and to determine which are the most relevant and accurate to perform in clinical practice for estimating the severity of preeclampsia. Twenty patients with mild preeclampsia, twenty patients with severe preeclampsia, ten eclamptic cases and ten normotensive pregnant women were included in the study. All cases and controls were with gestational age ranging between 28 and 38 weeks. All five markers increased and platelet count decreased in the severe preeclampsia and eclampsia groups. A highly significant negative correlation was found between platelet count and both P-selectin and Pf4 in the three studied hypertensive groups. Moving from mild to severe preeclampsia to eclampsia, vWF and Pf4 showed an increasingly abnormal results. Pf4 was the only haemostatic marker elevated in the mild preeclampsia group as compared with the normal pregnant group [P=0.000]. [1] Platelet activation may play an important role in the pathogenesis of preeclampsia; [2] Plasma levels of vWF and Pf4 could reflect the severity of this disease, [3] Pf4 appears to be an interesting marker for detecting early alterations in the haemostatic system in pregnancies complicated by preeclampsia. It seems that measurements of haemostatic markers in patients with preeclampsia may have prognostic value in determining the outcome of pregnancy in this pregnancy disorder. They offer possibilities of early assessment of therapeutic approaches aimed at limiting vascular endothelial damage and platelet activation