The Effects of Hydroxyurea on the Thrombin Generation and Microparticles in Sickle Cell Anemia Patients

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 2292-2292
Author(s):  
Patrick Van Dreden ◽  
Grigoris T Gerotziafas ◽  
Barry J Woodhams ◽  
Mourad Chaari ◽  
Robert Girot ◽  
...  
Keyword(s):  
Steady State ◽  
Sickle Cell ◽  
Thrombin Generation ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
No Production ◽  
Generation Process ◽  
Coagulation Activation ◽  
Significant Difference ◽  
Hydroxyurea Therapy

Abstract Abstract 2292 Introduction: The clinical course of sickle cell disease (SCD) is punctuated by episodic vascular occlusive events. The possibility that activation of the clotting system plays a contributory role in these complications is supported by abundant clinical data during both steady-state disease and pain crisis. Hydroxyurea therapy induces fetal haemoglobin, improves laboratory parameters and reduces acute clinical complications of SCD, but despite an abundance of evidence for coagulation and platelet activation, it remains incompletely defined whether these changes contribute to the reduced thrombin generation. This study is designed to evaluate coagulation profiles of patients with SCA in steady state and to determine whether hypercoagulable state is modified or not in patients on hydroxyurea therapy. Patients and Methods: We studied erythrocyte derived microparticles (Ed-MP) and platelet derived microparticles (Pd-MP) expressing or not expressing phosphatidylserine (PS) in patients with steady state SCD and we evaluated their specific procoagulant activity and their impact on thrombin generation process. A total of 92 steady state SCD patients were included in the study, of which 19 were under treatment with hydroyurea. The control group consisted of 30 healthy age and sex matched controls. Microparticles in whole blood were assessed using flow cytometry. Ed-MP and Pd-MP were identified using an anti-CD235 and CD41 monoclonal antibodies and annexin V. Thrombin generation in platelet poor plasma (PPL) was measured by CAT assay using PPP-reagent 5pM (Thrombinoscope, The Netherlands). Procoagulant phospholipid dependent activity in plasma was assessed by the Procoag-PPL assay (Diagnostica Stago, France). Thrombomodulin (TM) levels were measured by enzyme-linked immunosorbent assay (Elisa) Asserachrom thrombomodulin (Diagnostica Stago, Asnieres, France). Results: Hydroxyurea treated patients had lower counts of leukocytes, reticulocytes and platelets and an increased mean hemoglobin concentration as compared to non treated patients. Leukocyte and reticulocytes counts of treated patients were higher than those of controls. Platelets counts did not differ between treated and untreated patients. Patients on treatment with hydroxyurea had significantly lower levels of Ed-MP/PS+ and Ed-MP compared to untreated patients. The concentration of Pd-MP/PS+ and Pd-MP were not significantly different between hydroxyurea treated and non treated patients. The Ed-MP/PS+ showed a significant inverse correlation with Hb F (p<0.05). Thrombogram parameters, lag-time, ttPeak, Peak and MRI were significantly different between hydroxyurea treated patients and non treated patients. In hydroxyurea treated patients in contrast to the untreated ones no correlation was found between Ed-MP/PS+ and Ed-MP and parameters of thrombin generation. Among hydroxyurea treated patients 68% showed MRI levels higher than the UNL. Stratification groups of treated patients according to the levels of microparticles with Ed-MP/PS+ or Pd-MP/PS+ concentration higher than the UNL showed non significant difference compared to entirely group of patients. The PPL concentration was significantly lower in the SCD-treated patient compared to untreated patients (p<0.05). In contrast to platelet-derived-microparticles, the numbers of erythrocyte-derived-microparticles differed between patients with and without hydroxyurea during steady state. In patients treated with hydoxyurea, platelets were correlated with Ed-MP, Pd-MP with and without PS+ (p<0.05), but any of the others parameters showed one association. Procoagulant phospholipids and thrombomodulin were increased in SCD with and without hydroxyurea compared with controls group (p<0.05). Conclusion: Treatment with hydroxyurea result in decreases in plasma markers of thrombin generation, and may decrease coagulation activation by reducing PS expression on the surface of both RBCs and platelets in addition to being a NO donor hydro may also decrese haemostatic activation by its effect in decreasing the white blood cell count and particularly monocytes that express TF, furthermore the beneficial effects of hydroxyurea may be due to vasodilatationand decressed platelet and coagulation activation following NO production. Disclosures: Van Dreden: Diagnostica Stago: Employment. Gerotziafas:APHP: Employment. Woodhams:Diagnostica Stago: Employment. Chaari:APHP: Employment. Girot:APHP: Employment. Kartechi:APHP: Employment. Galea:APHP: Employment. Lionnet:APHP: Employment. Elalamy:APHP: Employment.

Steady State Sickle Cell Anemia Is Associated with Increased Formation of Erythrocyte-Derived Microparticles and Acceleration of Thrombin Generation.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 4001-4001 ◽  
Author(s):  
Mourad Chaari ◽  
Katia Stankovic ◽  
Vasso Galea ◽  
Francoise Robert ◽  
Amir Khaterchi ◽  
...  
Keyword(s):  
Steady State ◽  
Plasma Concentration ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Plasma Levels ◽  
Thrombin Generation ◽  
Conflicts Of Interest ◽  
Inverse Correlation ◽  
Control Group ◽  
Generation Process

Abstract Abstract 4001 Poster Board III-937 Introduction Patients with Sickle Cell Anemia (SCA) are at risk of thrombosis, but this clinical manifestation is variable and it is probably not associated with the frequency of vaso-occlusive crisis. Increased plasma levels of platelet- and erythrocyte-derived microparticles and hypercoagulability markers have been reported in steady state SCA patients. However, the link between hypercoagulability and SCA is not completelty understood. Aim of the study We determined erythrocyte-derived (Ed-MP) and platelet-derived microparticles (Pd-MP) levels in patients with steady state SCA. We studied their relationship with hemolysis markers and their impact on thrombin generation process. Materials and methods Consecutive out-patients with steady state SCA (n=78) and 20 healthy age and sex-matched controls were included. They were free of any acute episode of SCA for at least one month prior inclusion. Microparticles were assessed with standardized whole blood flow cytometry assay. Ed-MP and Pd-MP were identified using respectively anti-CD235a and anti-CD41 monoclonal antibody and annexine V. Thrombin generation (TG) in citrated platelet poor plasma was assessed with Calibrated Automated Thrombogram® (Stago, France) using PPP-reagent 5pM® (Thrombinoscope BV, Nederlands). The following TG parameters were analyzed: lag-time (LT), time to peak of thrombin (ttpeak), peak of thrombin (peak), mean velocity rate index of the TG (MRI) and endogenous thrombin potential (ETP). Results Mean patient age was 25±8 (range 17-58 ys). In the patients group, Ed-MP and Pd-MP, expressing or not phosphatidyl-serine (PS), were significantly increased compared to the control group. Thrombogram parameters were not significantly different in both groups (Table 1). There was a slight though significant inverse correlation between Ed-MP and both LT and ttpeak (r=-0.235, r=-0.315 respectively; p<0.05). Ed-MP levels were correlated with MRI increase (r=0.241; p<0.05). Ed-MP values were inversely correlated with Hb levels and well correlated with reticulocytes count (r=-0.427, r=0.520 respectively; p<0.05). No relationship was found between Ed-MP and ETP values. The sub-population of Ed-MP/PS+ (expressing PS) showed also an inverse correlation with both ttPeak (r=-0.315, p<0.05) and ETP (r=-0.236; p<0.05), and a positive correlation with MRI (r=0.306 ; p<0.05). Pd-MP concentration was inversely correlated with Hb levels (r=-0.273 ; p<0.05). Only Pd-MP/PS+ plasma concentration was slightly by significantly correlated with ETP (r=0.225; p=0,049). Conclusion Patients with steady state SCA presented a significant increase of Ed-MP and Pd-MP plasma levels which seems to be linked to haemolysis degree. Each type of microparticles had different impact on TG process. Ed-MP induced acceleration of TG kinetics without increase of ETP whereas Pd-MP/PS+ affected mainly ETP. However the increase of Ed-MP and Pd-MP plasma concentration does not appear to be by itself a sufficient condition to induce a significant increase of TG. Disclosures: No relevant conflicts of interest to declare.

FXIIa Differentially Regulates Thrombin Generation during Steady State and Vaso-Occlusive Crisis in Sickle Cell Mice

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 162-162 ◽  
Author(s):  
Erica M Sparkenbaugh ◽  
Camille Faes ◽  
Denis Noubouossie ◽  
Daniel K. Kirchhofer ◽  
András Gruber ◽  
...  
Keyword(s):  
Steady State ◽  
Mouse Model ◽  
Vascular Permeability ◽  
Sickle Cell ◽  
Plasma Levels ◽  
Thrombin Generation ◽  
Conflicts Of Interest ◽  
Control Group ◽  
Liver And Kidney

Abstract Sickle cell disease (SCD) is associated with chronic activation of coagulation. Previously, we demonstrated that inhibition of tissue factor (TF) attenuates thrombin generation (measured by plasma levels of thrombin-antithrombin complexes [TAT]) in a mouse model of SCD during steady state. Furthermore, we showed that neither inhibition of FXIIa-dependent activation of FXI (using 14E11 antibody) nor FXI deficiency reduces thrombin generation (TG) in sickle mice. In contrast, genetic deficiency of FXII or kininogen (HK) reduced plasma TAT levels. These data suggest that during steady state, FXIIa contributes to TG in sickle mice via activation of the kallikrein/HK pathway, but not FXI. In the present study, we further investigated the mechanisms of HK-induced TG at steady state, and increased TG observed during vaso-occlusive crisis (VOC). All experiments were performed using 4-5 month old Townes SS (sickle) and AA (control) mice. Kallikrein cleaves HK into HK fragments (HKFs) and bradykinin (BK). First, we investigated whether a BK-mediated increase in vascular permeability contributes to TG by exposing perivascular TF. This hypothesis was disproved by data demonstrating no difference in vascular permeability (measured by the extravasation of Evans blue in the heart, lung, liver and kidney) between AA (n=8) and SS (n=10) mice. HKFs were shown to induce leukocyte TF expression in vitro via binding to CD11b/CD18 (Mac-1). Therefore, we investigated whether Mac-1 inhibition affects TG in SS mice. AA and SS mice were treated with an inhibitory anti Mac-1 (M1/70) or IgG control antibody on days 0, 3 and 6 (i.p. 1 mg/kg) and TG was analyzed 1 day after the last injection. In the control group, SS mice demonstrated higher plasma TAT levels compared to AA mice (8.1±1.6 vs 4.2±0.6 ng/mL, n=10-11, p<0.05), but inhibition of Mac-1 significantly reduced plasma TAT levels in SS mice (4.6±0.7 ng/mL, n=11, p<0.05). These data suggest that HK might contribute to TG during steady state via Mac-1-dependent induction of monocyte TF. The steady state of SCD is interspersed with acute periods of VOC. Clinical data demonstrate that compared to the steady state, plasma levels of cell free DNA (cfDNA), activation of the contact system, and TG are further enhanced during VOC. To determine the mechanism of increased TG during VOC, we used the previously characterized mouse model of TNFα -induced VOC. Townes AA and SS mice were injected with recombinant TNFα (2 µg/g body weight) or the same volume of PBS, and plasma was collected 5 hours later. TNFα not only dramatically increased plasma levels of cfDNA in SS mice (14.78 ± 1.64 vs 679 ± 300 ng/mL; p<0.01), but also further increased plasma TAT levels compared to those observed in PBS-treated SS mice (2.9 fold, p<0.001, n=8). Importantly, there was a significant positive correlation between cfDNA and TAT in SS mice (r2 =0.65, p<0.001). Since cfDNA can activate FXII, we determined whether FXIIa-dependent activation of FXI contributes to TG during VOC. AA and SS mice received 14E11 or IgG control (4 mg/kg) 30 minutes before TNFα (2 μg/g) or PBS injection, and plasma TAT was assessed 5 hours later. Strikingly, 14E11 attenuated the increased TAT level in TNFα-treated SS mice, to the level observed in SS mice injected with PBS and IgG (IgG/SS/PBS: 9 ng/mL ± 1.8 vs. IgG/SS/TNF: 18.9 ± 3.6, p<0.001; 14E11/SS/TNF: 9.86 ± 0.72, p<0.05 vs. IgG/SS/TNF). We also determined if TF activity is required for the increased TG observed during VOC. Interestingly, inhibition of TF with an inhibitory 1H1 antibody (25 or 75 mg/kg injected i.p. 1 or 18 hours prior to TNFα, respectively) had no effect on the increased TG observed in TNFα treated SS mice. In aggregate, our data suggest that during the steady state of SCD, FXII-dependent TG is not FXI-dependent, but instead is mediated by a pathway involving HK, Mac-1 integrin and leukocyte TF. Furthermore, we propose that during VOC the massive release of cfDNA results in FXIIa-dependent FXI activation and enhances TG independently of TF. This study provides mechanistic insight into the initiators of TG in SCD. Moreover, it implicates FXIIa as a potential therapeutic target to reduce the prothrombotic state in SCD, during both steady state and VOC. Disclosures No relevant conflicts of interest to declare.

The acceleration of the propagation phase of thrombin generation in patients with steady-state sickle cell disease is associated with circulating erythrocyte-derived microparticles

2012 ◽  
Vol 107 (06) ◽  
pp. 1044-1052 ◽  
Author(s):  
Grigoris Gerotziafas ◽  
Patrick Van Dreden ◽  
Mourad Chaari ◽  
Vassiliki Galea ◽  
Amir Khaterchi ◽  
...  
Keyword(s):  
Steady State ◽  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Thrombin Generation ◽  
Generation Process ◽  
Cell Disease ◽  
Significant Acceleration ◽  
High Concentrations ◽  
The Impact ◽  
Functional Alteration

SummarySickle cell disease (SCD) is linked to hypercoagulability and is characterised by high concentrations of erythrocyte-derived microparticles (Ed-MPs). However, the impact of procoagulant cell-derived microparticles on the thrombin generation process remains unclear. We analysed the alterations of each phase of thrombin generation (TG) in relation to the concentration of erythrocyte- or platelet-derived microparticles (Ed-MPs and Pd-MPs) in a cohort of patients with steady-state SCD. We studied 92 steady-state SCD patients, 19 of which were under treatment with hydroxyurea, and 30 healthy age- and sex-matched individuals. TG was assessed by calibrated automated thrombogram. Ed-MP and Pd-MP expressing or not phosphatidylserine (PS) were determined by means of flow cytometry. Procoagulant phospholipid-dependent activity in the plasma was evaluated by the Procoag-PPL assay. Levels of thrombomodulin and haemoglobin in the plasma as well as red blood cell and reticulocyte counts were measured. SCD patients, independently of the administration of hydroxyurea, were marked by a significant acceleration in the propagation phase of TG which correlated with the Ed-MP/PS+ concentration. TG was significantly attenuated in hydroxyurea-treated patients. In conclusion, the acceleration of the propagation phase of TG, driven by Ed-MP/PS+, is a major functional alteration in blood coagulation in patients with steady-state SCD. Treatment with hydroxyurea, in addition to the regulation of haemolysis, lowers Ed-MPs and attenuates thrombin generation. The thrombogram could be a useful tool for the diagnosis of hypercoagulability and optimisation of the treatment in patients with SCD.

scholarly journals Circulating Erythrocyte-Derived Microparticles Are Associated with Coagulation Activation in Sickle Cell Disease

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 126-126 ◽  
Author(s):  
Eduard J. Beers van ◽  
M. C. Schaap ◽  
R.J. Berckmans ◽  
R. Nieuwland ◽  
A. Sturk ◽  
...  
Keyword(s):  
Steady State ◽  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Thrombin Generation ◽  
Absolute Difference ◽  
Glycophorin A ◽  
Cell Disease ◽  
Coagulation Activation ◽  
Factor Xi ◽  
The Absolute

Abstract Backgound: Sickle cell disease (SCD) is characterized by a hypercoagulable state involving multiple factors, including chronic hemolysis and circulating cell-derived microparticles (MPs). There is still no consensus on the cellular origin of such MPs and the exact mechanism by which they may support coagulation activation in SCD. Aim: In the present study, we aimed to analyze the origin of circulating MPs and their pro-coagulant phenotype in 25 consecutive SCD patients during painful crises and steady state. conditions Methods: MPs were identified by flowcytometry on basis of their size and density and on their ability to bind annexin V. Measurements were corrected for auto-fluorescence. Labeling with cell-specific monoclonal antibodies was corrected with isotype-matched control antibodies. For thrombin generation experiments patient derived MPs were reconstituted in defibrinated (reptilase-treated) normal pool (MP-free) plasma. For the inhibition experiments, the defibrinated plasma and the MPs were separately incubated with antibodies against coagulation factors VII or XI, or tissue factor pathway inhibitor (TFPI). Results: The majority of MPs originated from platelets (GPIIIa, CD61+: 86.1%) and erythrocytes (Glycophorin A, CD235+: 9.7%), and their numbers did not differ significantly between crisis and steady state. A distinct subset of transferrin receptor (CD71+)-exposing MPs was present, but neither MPs originating from monocytes (CD14+) nor endothelial cells (CD144+, CD146+, CD62E+) were detectable, and also no MPs exposing TF could be identified. The number of erythrocyte-derived MPs strongly correlated with plasma levels of hemolytic markers, i.e. hemoglobin (r=−0.58, P&lt;0.001) and lactate dehydrogenase (r=0.59;P&lt;0.001), von Willebrand factor as a marker of endothelial activation (r=0.44;P&lt;0.001), and D-dimer and prothrombin fragment F1+2 (r=0.52;P&lt;0.001 and r=0.59;P&lt;0.001, respectively) as markers of fibrinolysis and coagulation activation. In the thrombin generation test, the total amount of thrombin generated (represented by the area under the curve (AUC) of the thrombin generation curve) correlated with the total number of circulating MPs (R=0.63, P&lt;0.001). Thrombin generation was unaffected by pre-incubation with anti-human factor VII but increased slightly in the presence of anti-TFPI (16%; P=0.01) while the presence of anti-factor XI suppressed thrombin generation by about 50% (P&lt;0.001). The extent of this inhibition (as a percentage of thrombin generation without antibody) was significantly correlated with the circulating number of erythrocyte-derived MPs (r=0.50, P=0.023), but not with platelet-derived MPs or reticulocyte-derived MPs. Also the absolute difference in thrombin generation between the experiments with and without factor XI antibody correlated with the absolute number of glycophorin A+ MPs (r=0.55, P=0.002). Conclusion: We conclude that the procoagulant state in SCD is, at least in part, due to the procoagulant effects of circulating erythrocyte-derived MPs. Their relation to the ability of anti-factor XI to block thrombin generation in SCD patients suggests an important role of factor XI-dependent thrombin generation in these patients.

Levels of Prothrombin Fragment F1+2 in Patients with Hyperhomocysteinemia and a History of Venous Thromboembolism

1997 ◽  
Vol 78 (05) ◽  
pp. 1327-1331 ◽  
Author(s):  
Paul A Kyrle ◽  
Andreas Stümpflen ◽  
Mirko Hirschl ◽  
Christine Bialonczyk ◽  
Kurt Herkner ◽  
...  
Keyword(s):  
Venous Thromboembolism ◽  
Thrombin Generation ◽  
Oral Anticoagulant Therapy ◽  
Oral Anticoagulants ◽  
Control Group ◽  
Prothrombin Fragment ◽  
Hemostatic System ◽  
Significant Difference ◽  
System Activation ◽  
One Year

SummaryIncreased thrombin generation occurs in many individuals with inherited defects in the antithrombin or protein C anticoagulant pathways and is also seen in patients with thrombosis without a defined clotting abnormality. Hyperhomocysteinemia (H-HC) is an important risk factor of venous thromboembolism (VTE). We prospectively followed 48 patients with H-HC (median age 62 years, range 26-83; 18 males) and 183 patients (median age 50 years, range 18-85; 83 males) without H-HC for a period of up to one year. Prothrombin fragment Fl+2 (Fl+2) was determined in the patient’s plasma as a measure of thrombin generation during and at several time points after discontinuation of secondary thromboprophylaxis with oral anticoagulants. While on anticoagulants, patients with H-HC had significantly higher Fl+2 levels than patients without H-HC (mean 0.52 ± 0.49 nmol/1, median 0.4, range 0.2-2.8, versus 0.36 ± 0.2 nmol/1, median 0.3, range 0.1-2.1; p = 0.02). Three weeks and 3,6,9 and 12 months after discontinuation of oral anticoagulants, up to 20% of the patients with H-HC and 5 to 6% without H-HC had higher Fl+2 levels than a corresponding age- and sex-matched control group. 16% of the patients with H-HC and 4% of the patients without H-HC had either Fl+2 levels above the upper limit of normal controls at least at 2 occasions or (an) elevated Fl+2 level(s) followed by recurrent VTE. No statistical significant difference in the Fl+2 levels was seen between patients with and without H-HC. We conclude that a permanent hemostatic system activation is detectable in a proportion of patients with H-HC after discontinuation of oral anticoagulant therapy following VTE. Furthermore, secondary thromboprophylaxis with conventional doses of oral anticoagulants may not be sufficient to suppress hemostatic system activation in patients with H-HC.

scholarly journals Is there a role for selective vasodilation in the management of sickle cell disease?

Blood ◽  
1988 ◽  
Vol 71 (3) ◽  
pp. 597-602 ◽  
Author(s):  
GP Rodgers ◽  
MS Roy ◽  
CT Noguchi ◽  
AN Schechter
Keyword(s):  
Blood Flow ◽  
Steady State ◽  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Microvascular Obstruction ◽  
Controlled Study ◽  
Control Group ◽  
Cell Disease ◽  
Retinal Arteriolar

Abstract To test the hypothesis that microvascular obstruction to blood flow at the level of the arteriole may be significant in individuals with sickle cell anemia, the ophthalmologic effects of orally administered nifedipine were monitored in 11 steady-state patients. Three patients with evidence of acute peripheral retinal arteriolar occlusion displayed a prompt reperfusion of the involved segment. Two other patients showed fading of retroequatorial red retinal lesions. Color vision performance was improved in six of the nine patients tested. The majority of patients also demonstrated a significant decrease in the amount of blanching of the conjunctiva which reflects improved blood flow to this frequently involved area. Such improvements were not observable in a control group of untreated stable sickle cell subjects. These findings support the hypothesis that inappropriate vasoconstriction or frank vasospasm may be a significant factor in the pathogenesis of the microvascular lesions of sickle cell disease and, further, that selective microvascular entrapment inhibition may offer an additional strategy to the management of this disorder. We believe a larger, placebo-controlled study with nifedipine and similar agents is warranted.

scholarly journals Περιεγχειρητική κινητική αγγειορυθμιστικών παραγόντων σε ασθενείς με καρκίνο του μαστού

2014 ◽  
Author(s):  
Γεώργιος Γεωργίου
Keyword(s):  
Breast Cancer ◽  
Breast Adenocarcinoma ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Study Group ◽  
Rt Pcr ◽  
Altered Expression ◽  
Female Patients ◽  
Significant Difference ◽  
Circulating Levels

Βackground: angiogenesis is seen during the multiple stages of carcinogenesis, aswell as during the process of surgical wound healing, a fact which has led tosubstantial debate over the last decades about the potential impact of surgery upon thefinal outcome of ceratin patients treated for breast cancer.Aim: the present research aims at investigating the potential effect of surgery on theprocess of angiogenesis, by studying a number of factors that are related to the latter,in patients suffering from breast cancer before and after the time of the procedure,whilst comparing these results with those of patients that were operated on their breastfor non-malignant disease.Material-Methods: blood from 10 female patients with breast adenocarcinoma(Study Group) was collected via venipuncture before surgery (labeled as PRO), aswell as on post-operative day 3 (labeled as D3) and day 7 (labeled as D7). Moreover,blood samples were also taken from 6 female patients with fibroadenoma (ControlGroup) before surgery (PRO) and on day 3 afetr surgery (D3). These samples weremeasured for detection of circulating levels of three established angiogenesisbiomarkers using ELISA (Enzyme-Linked ImmunoSorbent Assay): VascularEndothelial Growth Factor-A (VEFG-A), Interleukin-8 (IL-8) and basic FibroblastGrowth factor (bFGF or FGF-2). In addition, circulating transcripts of 84 agiogenesirelatedgenes were determined using RT-PCR (Real Time Polymerase ChainReaction). The two groups of patients were firstly compared to each other regardingtheir results. Also, patients belonging to the Study Group were analized at differenttime points regarding surgery. Finally, the results were investigated againstclinicopathological data and patient outcome.Results: using ELISA we were able to detect increased levels of circulating VEGF-Aand IL-8 in the Study Group patients compared to the Control Group patientspreoperatively (p=0,0381 and p=0,0218 respectively), while for bFGF there was nostatistically significant difference documented. Surgery resulted in a significantincrease in VEGF-A levels on D3 (p=0,0389) and D7 (p=0,0172) as compared toPRO levels. Perioperative kinetics of IL-8 showed a mild trend towards increase,which, however, was not statistically significant. Postoperative levels of bFGF wereslightly increased on D3, but on D7 they were even lower than preoperative values(p=0,0205). Using RT-PCR certain differences between the Study Group and theControl Group were recorded regarding the circulating transcripts of a great numberof angiogenesis-related genes preoperatively: upregulation of VEGF-C, EGF, IL-8,FGF-1, SPHK1, NRP1, LAMA5, COL4A3, TEK, EFNA3, EFNB2. AKT1, ITGB3,THBS1, CCL11, TIMP3 and downregulation of CXCL10. Moreover, mastectomyinduced an altered expression in several key-genes in breast cancer patients:upregulation of THBS1, COL4A3, BAI1, ITGB3 and downregulation of EREG,SERPIFN1, CXCL9, CXCL10, IL1B, CCL2, CXCL1, HIF1A, NOTCH4. Conclusions: patients suffering from breast cancer have a different angiogenic profilein comparison to patients with fibroadenoma, as documented through their differencesin circulating levels of angiogenic factors. These levels are greatly changed after thesurgical procedure. VEGF showed a transient increase, while bFGF initially increasedbut only to finally decrease to levels that were even lower than the preoperative ones.Moreover, mastectomy promoted a shift in the expression pattern of a broad panel ofangiogenesis-related gene transcripts.

scholarly journals Suppression of transforming growth factor-β by mesenchymal stem-cells accelerates liver regeneration in liver fibrosis animal model

2021 ◽  
Vol 40 (1) ◽  
pp. 29
Author(s):  
Nur Anna C Sa’dyah ◽  
Agung Putra ◽  
Bayu Tirta Dirja ◽  
Nurul Hidayah ◽  
Salma Yasmine Azzahara ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Growth Factor ◽  
Liver Fibrosis ◽  
Transforming Growth Factor ◽  
Healing Process ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Significant Difference ◽  
T1 And T2

Introduction<br />Liver fibrosis (LF) results from the unregulated chronic wound healing process in liver tissue. Transforming growth factor-beta (TGF-β) is the major contributing cytokine of LF promotion through activation of quiescent hepatic stellate cells (HSCs) into myofibroblasts (MFs) and increased extracellular matrix (ECM) deposition such as collagen leading to scar tissue development. Mesenchymal stem cells (MSCs) have an immunomodulatory capability that could be used as a new treatment for repairing and regenerating LF through suppression of TGF-β. This study aimed to examine the role of MSCs in liver fibrosis animal models through suppression of TGF-β levels without scar formation particularly in the proliferation phase.<br /><br />Methods<br />In this study, a completely randomized design was used with sample size of 24. Male Sprague Dawley rats were injected intraperitoneally (IP) with carbon tetrachloride (CCl4), twice weekly, for eight weeks to induce LF. Rats were randomly assigned to four groups: negative control, CCl4 group, and CCL4 + MSC-treated groups T1 and T2, at doses of 1 x 106 and 2x106 cells, respectively. TGF-β levels were analyzed by enzyme-linked immunosorbent assay (ELISA). One-way ANOVA and a least significant difference (LSD) was used to analyse the data. <br /><br />Results<br />The TGF levels of LF rat models decreased on day 7 after MSC administration. The levels of TGF-β in both MSC groups T1 and T2 decreased significantly compared with the control group (p&lt;0.05). The TGF-β suppression capability of T2 was optimal and more significant than that of T1.<br /><br />Conclusion<br />MSCs can suppress TGF levels in liver fibrosis induced rats.

Investigating platelet-activating factor as a potent proinflammatory mediator in coronary atherosclerotic patients

2021 ◽  
Vol 67 (3) ◽  
pp. 1-4
Author(s):  
Shler Ghafoor Raheem
Keyword(s):  
Coronary Atherosclerosis ◽  
Platelet Activating Factor ◽  
Density Lipoprotein ◽  
High Sensitivity ◽  
Prognostic Indicator ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Surgical Specialty ◽  
Significant Difference ◽  
Hs Crp

The inflammatory reaction is one of the complications in patients with coronary atherosclerosis. This study aimed to determine the diagnostic value of platelet-activating factor (PAF) compared with high sensitivity C reactive protein (hs-CRP) in coronary atherosclerotic patients. Fifty patients with coronary atherosclerosis and 30 subjects with normal angiography were considered as the control group attending Cardiac Center-Surgical Specialty Hospital - in Erbil city / Iraq. The levels of PAF and hs-CRP were estimated quantitatively using a sandwich enzyme-linked immunosorbent assay and a particle-enhanced immune turbid metric assay, respectively. Lipid profiles and some hematological indexes were also used in this study. The levels of the inflammatory biomarkers of PAF and hs-CRP increased significantly in the patients group compared with controls (p<0.05). Although the patients group showed the highest level of low-density lipoprotein (LDL), the difference was not significant (p>0.05) compared with the healthy control. However, the incidence of risk factors such as smoking and obesity showed a significant difference (p<0.05) in the patients group. Additionally, the PAF level correlated positively and significantly with hs-CRP (p<0.05), and negatively with high-density lipoprotein (HDL) (p>0.05). Although hs-CRP was a valuable diagnostic marker for coronary atherosclerosis, the PAF level showed to be a better prognostic indicator than hs-CRP in coronary atherosclerosis patients.

scholarly journals P187 The significance of netrin-1 level in the clinical activity of ulcerative colitis, its association between TNF-α and IL-6

2020 ◽  
Vol 14 (Supplement_1) ◽  
pp. S232-S232
Author(s):  
H Korkmaz ◽  
K Fidan
Keyword(s):  
Ulcerative Colitis ◽  
Proinflammatory Cytokines ◽  
Activity Index ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Clinical Activity ◽  
Tnf Α ◽  
Significant Difference ◽  
And Control ◽  
Control Study

Abstract Background In this study, we investigated the importance of netrin-1 levels in ulcerative colitis (UC) in clinical activity of the disease, and its association with other proinflammatory cytokines IL-6 and TNF-α. Methods This study is a type of case–control study. Sixty-seven patients with UC (36 of them activation, 31 of remission) and 50 healthy controls were included in the study. UC patients; ‘Truelove Witts clinical activity index by remission (n = 31), mild activation (n = 21), moderate activation (n = 6) and severe activation (n = 9) were divided into groups. Netrin, IL-6 and TNF-α measurements in plasma samples were performed using enzyme-linked immunosorbent assay kit. Results Between the patient group and the control group; there was a statistically significant difference between netrin-1, IL-6, TNF-α, neutrophil, platelet (p &lt; 0.05 for all). The plasma netrin-1 mean of UC with severe activation group (139.21 ± 48.09 pg/ml) was statistically significantly higher than that of the mild activation (p = 0,037), remission group (p = 0,001) and control group(p = 0,011). The plasma netrin-1 mean of UC with moderate activation group was statistically significantly higher than that of the mild activation(p = 0,045) and remission group(p = 0,004). Conclusion Our results reveal that plasma netrin-1 levels have been shown to be associated with UC activation, similar to proinflammatory cytokines such as TNF-α and IL-6, in UC.

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