Circulating Biomarkers in Patients With Locally Advanced or Metastatic Renal Cell Carcinoma Treated With Everolimus in the Pre-nephrectomy Setting.

Many drugs are available in renal cell carcinoma (RCC), yet clinicians are still looking for predictive biomarkers of disease recurrence or progression supporting more personalised treatments. An assessment of circulating biomarkers over time was carried out in this French, open-label, single-arm, multicentre trial conducted in 25 patients with either locally advanced (n = 14) or metastatic RCC (n = 11) who received everolimus (10 mg daily) for 6 weeks prior to nephrectomy (NEORAD, NCT01715935). Circulating biomarkers, including circulating tumour cells, haematopoietic and endothelial cells, plasma angiogenesis and inflammatory markers were quantified at baseline, upon everolimus and post-nephrectomy. We assessed tumour burden, objective response rate upon RECIST1.1, disease-free survival (DFS) and progression-free survival (PFS). The correlation between circulating biomarkers was evaluated with multiple factor analysis and biomarker association with DFS/PFS by Cox regression. No objective response rate was obtained before nephrectomy. Upon everolimus, neutrophils, platelets and sVEGFR2 significantly decreased. We did not find any association between circulating biomarkers and DFS/PFS, but patients with the highest tumour burden at baseline had significantly higher plasma levels of interleukin-6, an inflammatory circulating biomarker, and lower levels of sVEGFR2, related to angiogenesis. Further understanding of the link between these circulating biomarkers could help to optimise drug combinations in RCC.

[Antiplatelet agents and transfusion]. / Antiplaquettaires et transfusion.

Antiplatelet agents are at risk for bleeding complications, the management of which differs depending on the clinical situation and on the antiplatelet agent itself. Neutralization of antiplatelets is sometimes necessary, most often leading to platelet transfusion, although the benefit of this strategy is poorly documented. In addition, if platelet transfusion corrects the platelet inhibition induced by aspirin and probably by clopidogrel and prasugrel, it does not neutralize ticagrelor, as a consequence of its pharmacological properties. The clinical benefit of platelet transfusion is limited, and the most recent studies are challenging it. However, it is indicated on a perioperative basis for surgeries with high hemorrhagic risk and is discussed in severe hemorrhages. The neutralization of ticagrelor is a concern and the antidote currently under development may be a solution. In all cases, other therapeutic solutions may be considered, such as administration of desmopressin, tranexamic acid or activated factor VII.

Prospective observational study of the effect of dual antiplatelet therapy with tranexamic acid treatment on platelet function and bleeding after cardiac surgery.

BACKGROUND: The bleeding impact of dual antiplatelet therapy (DAPT), aspirin and clopidogrel, maintained until coronary artery bypass graft surgery (CABG), is still a matter of debate. The lack of preoperative antiplatelet activity measurement and heterogeneity of antifibrinolytic protocols in prior studies make the conclusions questionable. The aim of this prospective study was to determine, after preoperative antiplatelet activity measurement, if the maintenance of DAPT until CABG increases bleeding in patients treated with tranexamic acid (TA). METHODS: This observational study included 150 consecutive patients, 89 treated with aspirin and 61 treated with DAPT, undergoing a first-time planned on-pump CABG with TA treatment. Antiplatelet activity was measured with platelet aggregation tests and quantification of VASP phosphorylation. Postoperative bleeding at 24 h was recorded and propensity score analysis was performed. RESULTS: Based on VASP assay, 54% of patients showed high on-clopidogrel platelet activity inhibition. Postoperative bleeding at 24 h increased by 22% in the DAPT group, compared with the aspirin group (680 [95% CI: 360-1670] vs 558 [95%CI: 267-1270] ml, P < 0.01), consistent with increased blood transfusion (21% vs 7%, P = 0.01); a higher incidence of mediastinitis did not reach statistical significance (15% vs 4%, P = 0.05). Bleeding correlated with the extent of clopidogrel antiplatelet effect, with the best correlation for the VASP assay. CONCLUSIONS: Maintenance of DAPT until the day of CABG in patients treated with TA, increased postoperative bleeding at 24 h in parallel with preoperative antiplatelet activity induced by clopidogrel.

[Thrombocytopenia associated with iron deficiency: a rare differential diagnosis of auto-immune thrombocytopenic purpura]. / Thrombopénie ferriprive : un diagnostic différentiel rare du purpura thrombopénique auto-immun.

INTRODUCTION: Iron deficiency is typically associated with microcytic anemia and thrombocytosis. It is a very uncommon cause of thrombocytopenia. CASE REPORT: A 17-year-old female presented with marked fatigue and dyspnea on exertion. Review of systems was only remarkable for abundant menstruations during the past two years. The hemogram revealed a profound microcytic anemia (4.4 g/dL, mean corpuscular volume [MCV] 49 fL) and a thrombocytopenia (33 G/L). Marked iron deficiency was also present: ferritinemia <3 µg/L. Investigations did not find any cause of iron deficiency anemia other than excessive menstrual loss. Bone marrow examination showed an increase number of megakaryocytes, compatible with an immune thrombocytopenia purpura. Iron supplementation completely normalized the platelet count within 48 hours. CONCLUSION: Iron affects thrombopoiesis. Because the number of megakaryocytes may then increase in the bone marrow, "iron deficiency thrombocytopenia" may be falsely diagnosed as immune thrombocytopenic purpura, leading to inappropriate corticosteroid therapy. Iron supplementation is the appropriate treatment of iron deficiency thrombocytopenia and allowed a rapid correction of the platelet count in all the 24 cases that have been previously reported with sufficient detail to be analyzed in the literature.

Perfusion defects after pulmonary embolism: risk factors and clinical significance.

BACKGROUND: Little is known about residual abnormalities after pulmonary embolism (PE). OBJECTIVES: To assess risk factors and the clinical significance of perfusion defects in patients with PE. PATIENTS/METHODS: Consecutive patients receiving at least 3 months of anticoagulant for an acute PE were included in a prospective cohort study. Ventilation/perfusion lung scan, echocardiography, 6-min walk test, thrombophilia and hemostatic variables were performed 6-12 months after PE. Perfusion defect was defined as a perfusion defect in at least two segments. RESULTS: Seventy-three out of 254 patients (29%) had perfusion defects during follow-up (median 12 months) and were more likely to have dyspnea, had a higher systolic pulmonary arterial pressure [39 mmHg (SD) (12) vs. 31 mmHg (8); P < 0.001] and walked a shorter distance during the 6-min walk test [374 m (122) vs. 427 m (99); P = 0.004]. Age [odds ratio (OR) 1.35; 95% confidence interval (CI), 1.11-1.63], the time interval between symptom onset and diagnosis (OR, 1.17; 95% CI, 1.04-1.31), pulmonary vascular obstruction at the onset of PE (OR, 1.34; 95% CI, 1.16-1.55) and previous venous thromboembolism (OR 2.06; 95% CI, 1.03-4.11) were independent predictors of perfusion defect after treatment of acute PE. Total tissue factor pathway inhibitor concentration was associated with perfusion defects. CONCLUSIONS: Perfusion defects are associated with an increase in pulmonary artery pressure (PAP) and functional limitation. Age, longer times between symptom onset and diagnosis, initial pulmonary vascular obstruction and previous venous thromboembolism were associated with perfusion defects.

Blood glucose levels in patients with metastatic renal cell carcinoma treated with sunitinib.

Sunitinib, a multitargeted tyrosine-kinase inhibitor, extends survival of patients with metastatic renal cell carcinoma (mRCC) and gastrointestinal stromal tumours. Between October 2005 and March 2007, we retrospectively reviewed blood glucose level variations associated with sunitinib therapy in patients treated for mRCC. Nineteen of the patients had type II diabetes. All 19 patients had a decrease in blood glucose level (mean 1.77 mmol l(-1)) after 4 weeks of treatment. This was followed by re-elevation in the 2-week rest period. After two cycles of sunitinib administration, two patients had stopped blood glucose-lowering drugs whereas five other patients had normalised their blood glucose level. On the basis of pre-clinical data, we hypothesise that several mechanisms could be involved in this process, such as capillary regression of pancreatic islets, IGF-1 modulation through HIF1-alpha or NF-kappaB activation. In addition, a decrease of glucose uptake in the context of concomitant gastrointestinal toxicity cannot be excluded. Glycaemic control should be carefully evaluated in diabetic patients treated with sunitinib, and routine monitoring is warranted.

Neoangiogenesis induced by progenitor endothelial cells: effect of fucoidan from marine algae.

Fucoidans--sulphated polysaccharides extracted from brown algae--could be beneficial in patients with ischemic diseases. Their antithrombotic and proangiogenic properties promote in animals, neovascularization and angiogenesis which prevent necrosis of ischemic tissue. In 1997, endothelial progenitor cells were first identified in human peripheral blood. They are recruited from bone marrow and contribute to neovascularization after ischemic injury. Mobilization of these cells in ischemic sites is an important step in new vessel formation. It is thought that the progenitors interact with endothelial cells, then extravasate and reach ischemic sites, where they proliferate and differentiate into new blood vessels. Although chemokines, cytokines and adhesion molecules are thought to be involved, the precise mechanism of progenitor mobilization is not fully understood. Recent studies suggest that stromal-derived factor 1 plays a critical role at several steps of progenitor mobilization. Given the role of proteoglycans within bone marrow, at the endothelium surface, and in growth factor and chemokine binding, fucoidans might influence the mobilization of endothelial progenitor cells and their incorporation in ischemic tissue. This review provides an update on circulating endothelial progenitors and their role in neovascularization. It focuses on recent advances in our understanding of interactions between these progenitor cells and exogenous sulphated polysaccharides, and their implications for understanding the fucoidan mechanism of action.

[Prognostic factors in chronic lymphocytic leukaemia: contribution of recent biological markers]. / Facteurs pronostiques de la leucémie lymphoïde chronique: apport des marqueurs biologiques récents.

Chronic lymphocytic leukemia (CLL) is the most common lymphoid hemopathy in elderly. Diagnosis of CLL is easily made with a full blood count and immunophenotyping, but there is an heterogeneity in clinical evolution. Until now, scheduling of treatment is based on Rai or Binet staging systems. These staging systems can not distinguish patients with a rapid evolution and thus who will need an earlier treatment. In order to detect these patients, it is useful to have some relevant markers to predict disease evolution. This article reviews recent biologic markers that can be used to evaluate long term prognosis of CLL patients.

[Microbial polysaccharides of marine origin and their potential in human therapeutics]. / Les polysaccharides microbiens d'origine marine et leur potentiel en thérapeutique humaine.

Bacterial polysaccharides offer fascinating potential applications for the pharmaceutical industry. Although many known marine bacteria produce exopolysaccharides (EPS), continuation in looking for new polysaccharide-producing micro-organisms is promising. Marine bacteria, isolated from deep-sea hydrothermal vents, have demonstrated their ability to produce in aerobic conditions, unusual EPS. With the aim of discovering biological activities, EPS presenting different structural features were studied. An EPS secreted by Vibrio diabolicus was evaluated on the restoration of bone integrity in experimental model and was demonstrated to be a strong bone-healing material. Another EPS produced by Alteromonas infernus was modified in order to obtain new heparin-like compounds. Unlike the native EPS, the resulting EPS presented anticoagulant properties as heparin. These EPS could provide biochemical entities with suitable functions for obtaining new drugs. They present original structural feature that can be modified to design compounds and improve their specificity.

Characterization, chemical modifications and in vitro anticoagulant properties of an exopolysaccharide produced by Alteromonas infernus.

A new low-molecular-weight 'heparin-like' component was obtained from an exopolysaccharide produced by a mesophilic strain found in deep-sea hydrothermal vents. Data concerning the structure of the native high-molecular-weight exopolysaccharide (10(6) g/mol, 10% sulfate content) are reported for the first time. Two depolymerization processes were used to obtain low-molecular-weight (24-35x10(3) g/mol) oversulfated fractions (sulfate content 20 or 40%). Nuclear magnetic resonance studies indicated that after sulfation (40%), the low-molecular-weight fraction obtained by free radical depolymerization was less sulfated in the 6-O-position than the fraction depolymerized by acid hydrolysis. The free radical depolymerized product also had sulfated residues in the 4-O-position and disulfated ones in the 2,3-O-positions. Moreover, the compounds generated by the free radical process were more homogeneous with respect to molecular mass. Also for the first time, the anticoagulant activity of the low-molecular-weight exopolysaccharide fractions is reported. When the fractions obtained after sulfation and depolymerization were compared with heparins, anticoagulant activity was detected in oversulfated fractions, but not in native exopolysaccharide. The free radical depolymerized fraction inhibited thrombin generation in both contact-activated and thromboplastin-activated plasma, showing a prolonged lag phase only in the contact-activated assay. Affinity co-electrophoresis studies suggested that a single population of polysaccharide chains binds to antithrombin and that only a subpopulation strongly interacts with heparin cofactor II.

Factors V leiden and II 20210A in patients with symptomatic pulmonary embolism and deep vein thrombosis.

PURPOSE: Factor V Leiden and factor II 20210A are inherited disorders of the clotting system that occur frequently in patients with deep vein thrombosis. We conducted this study to determine whether these factors are also common in patients with pulmonary embolism. SUBJECTS AND METHODS: We determined the prevalence of factor V Leiden and factor II 20210A in 773 consecutive patients with objectively documented symptomatic deep vein thrombosis or symptomatic pulmonary embolism, or with a combination of these disorders. RESULTS: Isolated symptomatic deep vein thrombosis occurred in 345 patients; isolated symptomatic pulmonary embolism occurred in 236; and both anomalies occurred in 192. Factor V Leiden was present in 21 (9%) of the patients with isolated symptomatic pulmonary embolism, in 30 (16%) with both manifestations, and in 63 (18%) with isolated symptomatic deep vein thrombosis (P = 0.007). Factor V Leiden was more common among patients with deep vein thrombosis (odds ratio [OR] = 2.1; 95% confidence interval [CI]: 1.2 to 3.7; P = 0.006) or both pulmonary embolism and deep vein thrombosis (OR = 1.8; 95% CI: 1.0 to 3.3; P = 0.07) than among patients with isolated pulmonary embolism. Factor V Leiden was less common in massive pulmonary embolism (5% [7 of 127]) than in submassive pulmonary embolism (13% [21 of 155], P = 0.03). We found no significant difference in the prevalence of factor II 20210A among the three groups. CONCLUSION: Factors V Leiden and II 20210A vary in prevalence among patients with pulmonary embolism and deep vein thrombosis, suggesting that the risk of pulmonary embolization may vary among patients who have different causes of venous thromboses.

Detection of phosphatidylserine surface exposure on human erythrocytes using annexin V-ferrofluid.

The asymmetric transbilayer distribution of phospholipids in the plasma membrane and the regulation of phosphatidylserine (PS) exposure at the cell surface of animal cells are of high physiological significance. It has been shown previously that annexin V is one of the most sensitive tools with which the presence of small amounts of PS on the outer surface of eukaryotic cells can be detected. We present here the covalent coupling of annexin V molecules to magnetic nanoparticles of maghemite. The resulting annexin V-ferrofluid is used in the magnetic separation of PS exposing cells, as illustrated for human erythrocytes modified in their phospholipid transbilayer asymmetry by the use of a calcium ionophore. Results on stored human erythrocytes and comparison with results obtained using iodinated and fluorescein-labeled annexin V are also presented.

Sickle cell disease: relation between procoagulant activity of red blood cells from different phenotypes and in vivo blood coagulation activation.

In the present study we examined if, among other mechanisms, the abnormal exposure of phosphatidylserine at the surface of sickle red blood cells (RBCs) contributes to the hypercoagulability which characterizes homozygous sickle cell disease (SCD). The question was addressed by comparison of the procoagulant properties of RBCs from subjects with various phenotypes (SS, SC and AS) that differ in clinical presentation. As previously reported, SS-RBCs accelerated the prothrombin activation by factor Xa, by decreasing the Km of the reaction compared to normal RBCs. SC-RBCs and AS-RBCs also promoted prothrombin activation although their procoagulant properties were milder compared to SS-RBCs. A significant increase of the thrombin-antithrombin complexes was observed in SS subjects. Prothrombin fragment 1+2 (F1+2) was elevated in half of the SS subjects, but the difference with controls did not reach significance. Elevated levels of thrombin-antithrombin complexes were observed in a number of SC (4/11) and AS (3/12) subjects, but the difference with controls was not significant. A significant correlation was observed between the plasma levels of thrombin-antithrombin complexes in the subjects with SS, AS and AA phenotypes, and the procoagulant properties of RBCs. Our results strongly suggest that the procoagulant properties which characterize SS-RBCs also affect SC-RBCs and AS-RBCs, and that exposure of phosphatidylserine by RBCs contributes to the hypercoagulable state observed in SCD.

Polymorphism in exon 10 of the human coagulation factor V gene in a population at risk for sickle cell disease.

In Caucasians, the R506Q mutation in exon 10 of the factor V gene (FV Leiden) confers an increased risk of thromboembolism. We have scanned this region of the gene for possible mutations in 450 subjects from populations at risk for sickle cell disease (SCD). The R506Q mutation was absent in subjects from sub-Saharan Africa, whereas its allelic frequency was 2.5% in the West Indies. Only one other substitution with no functional consequences in vitro (R485K) was found (32.4% allelic frequency) in sub-Saharan Africa. Thus, we found no mutations in exon 10 of the FV gene constituting an additional risk factor for thrombosis in SCD in sub-Saharan Africa. This suggests that the putative selective advantage conferred by R506Q does not exist in these populations, unless R485K has functional consequences in vivo. If further suggests that R506Q in American Africans is of Caucasian origin. Our data are the first to document ethnic variations in the frequency of the R485K polymorphism.

Phosphatidylserine exposure and aminophospholipid translocase activity in Rh-deficient erythrocytes.

Endogenous phosphatidylserine (PS) exposure and lipid transport activity have been investigated for seven unrelated cases of Rhnull erythrocytes. Endogenous PS exposure was measured by prothrombinase activity. Out of six cases studied, two Rhnull samples exhibited abnormal aminophospholipid exposure, as suggested by the measurement of a lower Km of factor Xa for prothrombin. Aminophospholipid translocase activity was measured through the transbilayer redistribution of spin-labelled analogues of phospholipids. Provided that incubation conditions allow the maintainance of intracellular ATP level, no difference was observed between Rhnull and control erythrocytes, clearly indicating that the aminophospholipid translocase and Rh polypeptides are different molecular species.