Plasma Levels Of bFGF Research Articles

The Plasma Levels of bFGF and Microbiological Evaluation of Cell Homing Procedure of Incomplete Teeth Apex Maturation in Dogs

The Plasma Levels of bFGF and Microbiological Evaluation of Cell Homing Procedure of Incomplete Teeth Apex Maturation in Dogs

The Effect of Aerobic-resistance Training on Plasma Levels of bFGF in Coronary Artery Disease After CABG

Background and Aim: Coronary Heart Disease (CAD) is one of the most important causes of mortality. Exercise activities after coronary artery bypass surgery increase the density of myocardial capillaries called angiogenesis and improve cardiovascular function. The aim of the present study was to determine the effect of aerobic-resistance training on plasma basic fibroblast grown factor levels in post-coronary artery bypass grafting patients. Methods & Materials: The sample consisted of 30 men aged 45-60 years who underwent bypass surgery. They were randomly divided into experimental (n=15) and control (n=15) groups. The experimental group performed aerobic resistance training for eight weeks (3 sessions per week) and the control group did not. Blood samples were taken 24 hours before the start of the first training session and 48 hours after the last training session while all subjects were fasting. In vitro ELISA method was used to measure BFGF. Independent and paired t-test and Shapiro-Wilk test were used to analyze the data at the significant level of 0.05 in SPSS v. 21. Ethical Considerations: This study was registered (Ethics Code: IR.IAU.NEYSHABUR.REC.1398.01) in the Ethics Committee of Islamic Azad University, Neishabour Branch, and with Clinical Trial Code: IRCTID:IRCT20191228045919N1. Results: Eight weeks of aerobic-resistance training significantly increased bFGF levels in the exercise group compared to the control group (P=0.002). Also, pre and post changes in the experimental group showed a significant increase in plasma bFGF levels (P=0.002), which was not significant in the control group (P=0.758). Conclusion: According to the results, it may be concluded that combined (aerobic resistance) exercise may increase angiogenesis and capillary density in post-CABG patients by increasing plasma bFGF levels

Integrative biomarker analyses indicate etiological variations in hepatocellular carcinoma

The purpose of this study was to determine whether biomarkers from baseline plasma and archival tissue specimens collected from patients enrolled in the EVOLVE-1 trial - a randomized phase 3 study of everolimus in hepatocellular carcinoma (HCC) - were associated with prognosis, etiology or ethnicity. Circulating plasma levels of bFGF, PLGF, VEGF, VEGF-D, c-Kit, collagen IV, sVEGFR1 and VEGFR2 were measured by ELISA (N=503). Protein levels of IGF-1R, c-Met, mTOR, Tsc2 were assayed by immunohistochemistry (N=125). Genomic DNA sequencing was conducted on a panel of 287 cancer-related genes (N=69). Patients with baseline plasma concentrations of VEGF or sVEGFR1 above the cohort median had significantly shorter overall survival. These plasma biomarkers retained prognostic significance in a multivariate Cox regression model with geographic region, macroscopic vascular invasion and alpha fetoprotein AFP levels. Membranous c-Met protein levels were significantly lower for Asian patients, as well as for hepatitis B viral etiology. The prevalence of genetic changes were similar to previous reports, along with a trend towards higher PTEN and TSC2 mutations among Asians. The angiogenesis biomarkers VEGF and sVEGFR1 were independent prognostic predictors of survival in patients with advanced HCC. Potential differences in c-Met and mTOR pathway activation between Asian and non-Asian patients should be considered in future clinical trials. Our study demonstrates that circulating angiogenesis biomarkers can predict the survival outcome in patients with advanced hepatocellular carcinoma independent of the clinical variables. There is etiology and ethnicity variation in molecular pathway activation in hepatocellular carcinoma, which should be considered for future clinical trial design of targeted therapy. NCT01035229.

Evaluation of plasma basic fibroblast growth factor (bFGF) in primary knee osteoarthritis patients

Aim of the workThe aim of this study was to investigate plasma basic fibroblast growth factor (bFGF) levels in patients with primary knee osteoarthritis (KOA) and to correlate it with physical performance, functional status and radiological severity. Patients and methodsSixty patients with primary KOA and 30 healthy individuals were recruited into this study. Measures of physical performance were assessed using Chair Stand Test, Stair Climb Test and Six-Minute Walk Test. Functional assessment was carried out using the Western Ontario and McMaster Universities (WOMAC) index. KOA severity was determined using X-ray evaluated according to the Kellgren–Lawrence (KL) grading and all underwent sonographic examination. bFGF concentrations in plasma were determined. ResultsThe mean age of the KOA patients was 53.67±7.99years, female:male 4:1 and disease duration of 4.17±1.74years. KOA was bilateral in 71.7% and knee effusion was present in 20.39%. The WOMAC index was 53.78±13.7. Plasma bFGF levels in KOA patients (43.82±20.18pg/ml) were significantly higher than in controls (12.40±9.12pg/ml) (p< 0.001). bFGF significantly correlated the KL radiographic grading (r=0.31, p<0.027) and negatively with cartilage thickness of medial and lateral femoral condyles (r=−0.38, p< 0.006). ConclusionsPlasma bFGF levels were significantly increased in OA patients, and these elevated levels were significantly correlated with the degree of radiographic severity evaluated by KL grading scale and cartilage degeneration evaluated by ultrasound. These findings indicate that bFGF levels may be a monitor of disease severity and could play an essential part in the pathophysiology of degenerative process in OA.

Effect of Coronary Artery Bypass Grafting on the Plasma Levels of VEGF, bFGF, Copper and NO as Angiogenic Biomarkers in Chronic Ischemic Heart Disease Patients

Objectives: Myocardial ischemia offers paradoxical protection from ischemic necrosis by the development of coronary collateral circulation through the process of angiogenesis. The current study aimed to assess the plasma levels of vascular endothelial growth factor basic fibroblast growth factor nitric oxide and copper as angiogenic biomarkers to evaluate their role in the development of coronary collateral circulation in patients with chronic ischemic heart disease. Also, to evaluate the angiogenic modulator effects of coronary artery bypass grafting (CABG) via repeated measurements of such biomarkers 3 months postoperative. Methods: This case control and observational study involved 25 ischemic heart disease male patients candidate for elective coronary artery bypass grafting who admitted at the department of cardiothoracic surgery, faculty of medicine, Assiut University and 25 healthy age matched males as control group. Spectrophotometric assay of NO and copper and ELISA assay of VEGF and bFGF plasma levels were done. Results: Significant higher plasma VEGF, bFGF, copper and NO levels preoperatively when compared with the control group (p<0.01 for all) with significant lowering of their plasma levels when repeatedly measured three months following the CABG (p<0.01 for all). Conclusions: Our findings revealed that plasma levels of VEGF, bFGF, copper and NO have been significantly decreased in patients with chronic myocardial ischemia following CABG, thus, proving the proper coronary reperfusion effect of CABG with decrease the demand for the angiogenesis in the ischemic myocardium. There was a positive correlation between the plasma levels of VEGF and NO.

Correlation Between Clinical Responses and Immune Characteristics in Patients with Relapsed CLL Treated with Ofatumumab and Lenalidomide

Chronic lymphocytic leukemia (CLL) is characterized by phenotypical and functional alterations of different host immune components. Lenalidomide is an immunomodulatory agent which exerts both a direct antitumor effect and a pleiotropic activity on the immune system. Single-agent lenalidomide is active in patients with relapsed and refractory CLL, and the addition of an anti-CD20 monoclonal antibody to lenalidomide has shown synergism in vitro, as well as improved clinical responses in vivo.We designed a phase II study with the aim of determining the efficacy and tolerability of combined ofatumumab and lenalidomide for the treatment of relapsed or refractory CLL. We also wanted to investigate whether immune system and tumor microenvironment characteristics, such as circulating T and NK cell number and function, and plasma levels of chemokines, cytokines, and angiogenic factors could be correlated with the response to treatment and could be modulated during the course of the therapy. Ofatumumab was administered at a dose of 300 mg on day 1, 1000 mg on days 8, 15, and 22 during course 1, 1000 mg on day 1 during courses 3-6, and once every other course on day 1 during courses 7-24 (28-day courses). Lenalidomide (10 mg daily) was started on day 9 and continued for as long as a clinical benefit was observed.Thirty-four patients were enrolled. The overall response rate was 71%. Eight patients (24%) achieved a complete remission (CR) or CR with incomplete recovery of blood counts, including 9% with minimal residual disease-negative CR. The median progression-free survival was 16 months, and the estimated 5-year survival was 53%. The most common treatment-related toxicity was neutropenia (grade >2 in 18% of the 574 patient-courses). The most frequent infectious complications were pneumonia and neutropenic fever (24% and 9% of patients, respectively).For the correlative studies, peripheral blood mononuclear cells and plasma samples were collected at baseline and after 3, 6, 9, and 12 months of therapy. Lymphocyte subsets were analyzed by flow cytometry. T and NK cell function was evaluated on 9 patients based on the expression of CD107a, IFNγ, TNFα, and IL-2 after the exposure to CD3/CD28 magnetic beads and K562 cell line, respectively. Chemokines, cytokines, and angiogenic factors were quantified through a multiplex bead-based immunoassay. Thirty-four age-matched healthy donors (HD) were used as controls. Treatment with ofatumumab and lenalidomide decreased the number of circulating T and NK cells. The greatest decrease was observed after 3 months of therapy as compared to baseline (p≤0.02 for all comparisons). We found that the baseline number of CD4+ T cells and CD57+ CD56+ NK cells was higher in CR patients as compared to non responder (NR) patients (median absolute number 1595/μl vs. 715/μl, p=0.04, and 358/μl vs. 70/μl, p=0.02). We also compared baseline T and NK cell populations of CR patients to HD, and found that the former had significantly higher numbers of CD16+ CD56+ and CD57+ CD56+ NK cells (median 356/μl vs. 80/μl, p=0.03, and 358/μl vs. 29/μl, p=0.0008). Interestingly, baseline numbers of CD16+ CD56+ and CD57+ CD56+ NK cells were comparable between NR patients and HD. We also observed that patients who achieved CR had significantly better T and NK cell function at baseline, whereas T and NK cell function was impaired in patients who only had partial or no response. Plasma levels of bFGF, VEGF and IFN-γ were significantly decreased at month 3 compared to baseline in all patients (p≤0.02 for all comparisons). Finally, significantly lower levels of circulating CCL2, CCL3, CCL4, IFNα, and IL-12p70 were detected during treatment in patients who achieved CR, compared to NR (p<0.05 for all comparisons).In conclusion, the combination of ofatumumab and lenalidomide is a well tolerated regimen that induces durable responses in the majority of patients with relapsed CLL, and sustained MRD-negative CR in some. A higher number and preserved functional activity of T and NK cells were found in responding patients at baseline, as compared to NR, underscoring the importance of a competent immune system in supporting the effectiveness of this type of treatment, and suggesting a need to further investigate these immune features as markers for prediction of response to therapy. DisclosuresFaderl:Celgene Corp.: Other: Advisory Board. Wierda:Celgene Corp.: Consultancy; Glaxo-Smith-Kline Inc.: Research Funding. Rezvani:Pharmacyclics: Research Funding. Burger:Pharmacyclics LLC, an AbbVie Company: Research Funding. Keating:Glaxo-Smith-Kline Inc.: Other: Advisory board; Celgene Corp.: Consultancy.

Elevation of plasma basic fibroblast growth factor after nocturnal hypoxic events in patients with obstructive sleep apnea syndrome

Obstructive sleep apnea syndrome (OSAS) is associated with recurrent nocturnal hypoxia during sleep; this hypoxia has been implicated in the pathogenesis of cardiovascular complication. However, a useful soluble factor that is sensitively correlated with OSAS severity for the diagnosis remains unidentified. We hypothesized that systemic levels of basic fibroblast growth factor (bFGF), a hypoxia-induced cytokine, were affected by nocturnal hypoxemia in OSAS patients, and we assessed whether the degree of change in the plasma bFGF concentrations before and after nocturnal hypoxia is correlated with the severity of OSAS. Thirty subjects who had suspected OSAS and had been investigated by nocturnal polysomnography (PSG) were enrolled. Plasma bFGF and vascular endothelial growth factor (VEGF) concentrations the night before PSG and the next morning were measured by sandwich enzyme-linked immunosorbent assay. Correlations between the changes in these factors and hypoxia-associated parameters for OSAS severity were analyzed. Patients with OSAS had significantly elevated levels of plasma bFGF but not VEGF and hemoglobin after rising. The degree of change in bFGF concentrations after nocturnal apnea episodes was significantly correlated with diagnostic parameters for OSAS severity. The change in plasma bFGF levels is associated with the degree of hypoxic state in OSAS patients, implying that bFGF might be a useful soluble factor for evaluating OSAS severity.

Relationship between biomarkers and everolimus efficacy in the phase III RECORD-1 trial of patients with metastatic renal cell carcinoma (mRCC).

352 Background: Compared with placebo, everolimus provided significant improvement in median PFS in the RECORD-1 study of VEGFr-TKI-refractory mRCC. To investigate the role of angiogenesis pathway molecules as potential biomarkers of everolimus efficacy in RECORD-1, plasma levels of sVEGFR-2, VEGF-A, and bFGF were estimated. Methods: In addition to best supportive care, patients received everolimus 10 mg/day (n = 277) or placebo (n = 139). Placebo patients who progressed were offered everolimus. Predose blood samples were collected on day 1 of the first four 28-day treatment cycles. Plasma levels of sVEGFR-2, VEGF-A, and bFGF were assessed by ELISA. Effect of treatment over time on each biomarker was assessed by a mixed effects model. Hazard ratios (HR) for prognostic effects were obtained using log baseline biomarker values as continuous variables in a stratified Cox proportional hazards model. Results: Plasma levels of sVEGFR-2, VEGF-A, and bFGF were available for 45%, 45%, and 39% of everolimus patients and 50%, 50%, and 45% of placebo patients. Patients with biomarker data had baseline characteristics similar to those of the overall population. Mean baseline levels (pg/mL) of sVEGFR-2, VEGF-A, and bFGF were similar for everolimus (8945, 245, and 8, respectively) and placebo (8985, 253, and 13, respectively). Everolimus significantly improved median PFS over placebo irrespective of baseline levels of the analyzed biomarkers (p &lt; 0.001), indicating they are not predictive of everolimus efficacy. Prolonged PFS in the biomarker population was associated with lower VEGF-A baseline level (HR, 1.27; 95% CI, 1.03-1.57; p = 0.028), suggesting VEGF-A may be prognostic for mRCC. Compared with placebo, everolimus significantly reduced bFGF (p = 0.0095) and sVEGFR-2 (p&lt; 0.001) levels over the time course of the study; no effect on VEGF-A levels was observed. Conclusions: Everolimus significantly improved PFS compared with placebo, regardless of baseline biomarker levels. Lower VEGF-A levels may be a potential prognostic factor for longer PFS. Everolimus treatment significantly downregulated plasma levels of bFGF and sVEGFR-2 from baseline. Clinical trial information: NCT00410124.

In Vitro and in Vivo Evidence of an Anti-Angiogenic Effect of Lenalidomide in Chronic Lymphocytic Leukemia

AbstractAbstract 1782The exact mechanism of anti-tumor activity of the immunomodulatory drug lenalidomide remains undefined, but it implies the activation of immune effector cells. Little is known concerning the anti-angiogenic properties of lenalidomide in chronic lymphocytic leukemia (CLL) patients.A total of 27 relapsed/refractory CLL patients were treated within a multicenter Phase II trial. Peripheral blood mononuclear cells and plasma samples were collected before and during treatment. Lenalidomide treatment schedule began with 5 mg daily doses and increased by 5 mg daily every two weeks, up to 25 mg daily or maximum tolerated dose. Several angiogenesis-related cytokines were measured by ELISA in plasma. Circulating endothelial cells (CEC), endothelial progenitor cells (EPC), apoptotic CEC (APO-CEC), and activated CEC (aCEC) were evaluated by flow cytometry. In vitro experiments were performed on CLL cells purified from peripheral blood by using CD19Microbeads. Cells were cultured alone (CLL only) or on a EC monolayer (HC condition).First, CLL cells from 9 patients treated in vitro with up to 10 μmol/L of lenalidomide did not show any direct cytotoxic effect. However, an anti-leukemic effect was observed in vivo in our cohort of heavily pre-treated CLL patients. We found a reduction of white blood cell count (WBC, 1×109/L) from 39.9±8.0 before treatment to 15.2±5.2 and 16.0±5.2 after 4 and 12 months of treatment respectively and a decrement of absolute number of lymphocytes from 25.6±6.4 before treatment to 7.9±2.7 and 5.4±2.4 after 4 and 12 months of treatment (p<0.05, Wilcoxon test). To evaluate whether lenalidomide exerts its anti-CLL effect disrupting leukemic cross-talk with endothelial cells, we co-cultured CLL cells (n=9) on EC layer in presence or absence of increasing doses of lenalidomide. All CLL cells cultured on EC layer displayed a significant survival advantage (mean viability, 16% vs. 47% in CLL alone and HC) that was reduced by lenalidomide treatment (28%) (p<0.001).We then evaluated the effect of lenalidomide treatment on circulating endothelial cells in treated CLL patients (n=27). We found that the absolute number of CEC and EPC decreased from 8.2±1.5/μL and 4.7±1.0/μL pre-therapy to 2.1±0.6/μL and 1.0±0.5/μL after 4 months respectively (p=0.002 for CEC and p=0.01 for EPC). Moreover, the percentage of APO-CEC increased from 51% to 84% of the total CEC (p<0.0001). No differences were found in the aCEC subset during treatment. Moreover, we examined the effect of lenalidomide treatment on a panel of angiogenesis-related factors. The plasma levels of vascular endothelial growth factor (VEGF) and thrombospondin-1 (THBS1) showed a significant decrease by month 4 compared to pre-treatment levels (VEGF, 77±13 to 43±8 pg/mL, p=0.014; THBS1, 662±80 to 416±75 ng/mL, p=0.01). Angiogenin plasma levels increased after 4 months of treatment from 441±16 ng/mL to 460±21 ng/mL (p=0.025). No significant differences were detected in plasma levels of basic fibroblast growth factor (bFGF,) and Angiopoietin-1 (Ang1). Angiopoietin-2 (Ang2) showed an increase during the first phase of treatment from 5183 to 8126 pg/mL (p=0.001).Furthermore, patients were grouped under the clinical classification of stable/progressive disease (not responders, NR n=6) and partial response (responders, R n=10). There was an Increased percentage of aCEC detected in NR patients (54% vs. 74% in R and NR at baseline respectively, p=0.039). We also found that VEGF plasma levels significantly decreased during treatment exclusively in the R subset (83±26 pg/mL at baseline to 26±6 pg/mL after 4 months, p=0.007). Moreover, bFGF plasma levels showed a strong decrease in R patients from 168±105 pg/mL at baseline to 41±11 and 19±8 pg/mL after one week and 4 months of treatment (p=0.017 and p=0.005 respectively), whereas increased levels were measured in NR patients from 120±58 pg/mL at baseline to 426±195 and 234±147 pg/mL.Our study provides evidence that the mechanism of action of lenalidomide in CLL patients is mediated not only through the activation of immune effector cells, but also through the modulation of angiogenesis-related factors and disruption of CLL cross-talk with endothelial cells. Of interest, we identified plasmatic factors (VEGF and bFGF) that are correlated with clinical response to lenalidomide. Disclosures:Off Label Use: Lenalidomide, a thalidomide analogue, is an immunomodulatory drug (IMiD) with antitumoural activity reported in various malignant disorders including multiple myeloma and myelodysplastic syndrome (MDS). At preclinical level, lenalidomide has shown to decrease the production of several prosurvival cytokines. This drug is also reported to modulate an effector cell immune response through the activation of T and natural killer (NK) cells, inducing apoptosis directly on tumour cells. Currently available data indicate that Lenalidomide is active also in heavily pre-treated CLL patients with unfavourable prognostic factors. However, in order to optimize the therapeutic index of Lenalidomide treatment in CLL patients, it will be necessary to identify features of tumour cells that differ between responder and non responder patients. Hence, we reported results from a multicenter, phase II study designed in order to identify potential predictive factors correlating with response to Lenalidomide treatment in relapsed/refractory CLL. Marasca:CELGENE CORPORATION: Honoraria, Research Funding.

842P - Biomarkers of Everolimus Efficacy in Patients with Metastatic Renal Cell Carcinoma (MRCC): Analysis of the Phase III Record-1 Trial

ABSTRACT Background In RECORD-1, everolimus significantly increased median PFS (primary end point) over placebo in VEGFr-TKI-refractory patients with mRCC. We investigated angiogenesis pathway molecules, sVEGFR-2, VEGF-A, and bFGF, in plasma as potential biomarkers of everolimus efficacy in RECORD-1. Material and methods Patients received everolimus 10 mg daily (n = 277) or placebo (n = 139), both with BSC; placebo patients could cross over to everolimus at disease progression. Pre-dose blood samples were collected on day 1 of the first four 28-day treatment cycles; plasma levels of sVEGFR-2, VEGF-A, and bFGF were assessed using ELISA. A mixed effects model was used to assess treatment effect over time on each biomarker. Hazard ratios (HR) for prognostic effects were obtained using log baseline biomarker values as continuous variables in a stratified Cox proportional hazards model. Results Plasma values for sVEGFR-2, VEGF-A, and bFGF were available for 45/45/39% of everolimus patients and 50/50/45% of placebo patients. Baseline characteristics of patients with biomarker data were similar to the overall population. Mean log baseline values for sVEGFR-2, VEGF-A, and bFGF were similar for both arms; 9.1/5.1/1.6 for everolimus and 9.1/5.2/1.8 for placebo, respectively. Median PFS was significantly improved with everolimus vs placebo, regardless of baseline levels of any of the biomarkers analyzed (P Conclusions Everolimus provided significant clinical benefit over placebo, regardless of baseline biomarker levels. However, lower VEGF-A level was seen as a potential prognostic factor for longer PFS. Plasma levels of bFGF and sVEGFR-2 were significantly down-regulated from baseline by everolimus treatment. Disclosure S. Oudard: Stephane Oudard received honoraria from Bayer, Novartis, Pfizer, Roche, and Sanofi-Aventis. B. Escudier: Bernard Escudier has received honorarium from Novartis, Pfizer, GSK, Aveo, and Bayer. J. Thompson: John Thompson received clinical study support from Novartis. V. Grunwald: Viktor Grunwald served as consultant to Roche, Bayer, Novartis, Pfizer, GlaxoSmithKline, and Aveo/Astellas, received honoraria from GlaxoSmithKline, Novartis, and Pfizer, and received research funding from Pfizer and GlaxoSmithKline. S. Bracarda: Sergio Bracarda has served as advisor for Pfizer, Bayer-Schering, GlaxoSmithKline, Novartis, Aveo/Astellas, Boheringer-Ingelheim, Johnson & Johnson, and Sanofi-Aventis and has received speaker fees from Pfizer, Novartis, and Sanofi-Aventis. A. Panneerselvam: Ashok Panneerselvam is an employee of Novartis Pharmaceuticals Corporation. S. Gogov: Sven Gogov is an employee of Novartis Pharma AG. D. Chen: David Chen is an employee of Novartis Pharmaceuticals Corporation. R.J. Motzer: Robert Motzer has received research funding from Novartis. All other authors have declared no conflicts of interest.

Plasma biomarker analysis of a phase I trial of ASA404 (vadimezan) in combination with paclitaxel and carboplatin in Japanese patients with non-small cell lung cancer (NSCLC).

e18035 Background: ASA404 (vadimezan, DMXAA) is a flavonoid, non-tubulin-binding tumor-vascular disrupting agent (Tumor- VDA) that causes irreversible tumor vasculature disruption and extensive hemorrhagic necrosis of the tumor core. In combination with paclitaxel and carboplatin (P/C), ASA404 increased the median overall survival of previously untreated NSCLC patients from 8.8 to 14.0 months in a phase II clinical trial. Methods: A phase I trial assessed the safety profile and tolerability of three dose levels of ASA404 (600, 1,200, and 1,800 mg/m2) administered in combination with P/C (P, 200 mg/m2; C, AUC 6) every 3 weeks in Japanese patients with NSCLC. A panel of plasma biomarkers of angiogenesis and cytokines were evaluated to investigate the pharmacodynamic effect and mechanism of action (MOA) of ASA404. Plasma VEGF and basic FGF (bFGF) were measured by multiplex assays using the Meso-Scale Discovery platform. Plasma IL-8 and macrophage chemotactic protein (MCP)1 were analyzed by Luminex beads platform. Plasma 5-hydroxyindole-3-acetic acid (5HIAA), a by-product of platelet activation and putative biomarker for ASA404's vascular disruption activity, was analyzed by tandem mass spectrometry. Results: Four-hours after ASA404 infusion at Cycle 1 Day 1, plasma 5HIAA levels increased in a dose-dependent manner by 15%, 111%, or 137% compared with pre-ASA404 treatment level in the 600 (n = 3), 1,200 (n = 6) or 1,800 (n = 6) mg/m2 groups, respectively. Plasma VEGF levels decreased by 5 to 50% after 4 h and 24 h of ASA404 infusion. Plasma bFGF levels decreased by 55 to 88% after 24 h ASA404 infusion in the three ASA404 dose groups. Furthermore, over 50% induction of plasma IL-8 and MCP-1 were observed after 24 h infusion of ASA404. Conclusions: ASA404 caused dose-dependent induction of plasma 5HIAA levels. Unlike anti-angiogenic compounds such as sunitinib, sorafenib or bevacizumab which upregulate systemic VEGF, ASA404 did not appear to cause a systemic VEGF elevation. Induction of plasma inflammatory cytokines (IL-8, MCP-1) after ASA404 treatment may represent ASA404 MOA and amplify the direct antivascular effect of ASA404. Author Disclosure Employment or Leadership Position Consultant or Advisory Role Stock Ownership Honoraria Research Funding Expert Testimony Other Remuneration Novartis Novartis

Correlation of electroretinography b-wave absolute latency, plasma levels of human basic fibroblast growth factor, vascular endothelial growth factor, soluble fatty acid synthase, and adrenomedullin in diabetic retinopathy

Background We investigated the b-wave latency of electroretinogram (ERG), human basic fibroblast growth factor (b-FGF), vascular endothelial growth factor (VEGF), soluble fatty acid synthase (s-Fas), and adrenomedullin (ADM) in diabetic retinopathy. Patients and Methods Thirty control and 60 type II diabetic women (mean age 45±3.9 years, duration of diabetes 10.1±2.1 years) were investigated. Diabetics without complications (Group II) and with retinopathy (Group III) were diagnosed depending on clinical findings, abnormal fundus examination, and ERG. Plasma levels of b-FGF, VEGF, s-Fas, and ADM were measured. Results ERG showed a significant increase of b-wave absolute latency, plasma b-FGF, VEGF, s-Fas, and ADM in diabetic retinopathy ( P<.05). A positive correlation was found between b-wave latency and VEGF and s-Fas, and a negative correlation with b-FGF and ADM. Conclusion This study elucidates the causative role of VEGF and s-Fas in diabetic retinopathy. VEGF may potently promote growth of endothelial cells and formation of new vessels implicated in proliferative retinopathy. s-Fas could be involved in advancement of apoptotic changes in retinopathy and high levels of b-FGF, and ADM may be compensatorily neuroprotective and vasculoprotective. The results showed that diabetic retinopathy is the result of multiple factors, so it is optimistic to believe that reversing VEGF or s-Fas will halt retinopathy, targeting multiple mechanisms simultaneously by administering combination treatments of VEGF antagonists; antiapoptotic drugs together with b-FGF and/or ADM may be prospective.

Prognostic value of blood levels of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in advanced non-small cell lung cancer (NSCLC) patients

7196 Background: VEGF and bFGF are among the most important angiogenic factors. We have explored these angiogenesis mediators in plasma and its prognostic significance in advanced NSCLC. Methods: Were enrolled 451 patients with advanced NSCLC, stages IIIB and IV and treated with cisplatin and docetaxel. Blood was collected before chemotherapy. Plasma VEGF and bFGF levels were assessed by commercial ELISA (sensitivity 5 pg/ml). In parallel plasma from 32 age and gender-matched controls was used. Results: Median age was 61 years (35–82) and 84% were males. 99% had performance status 0–1. 84% were in stage IV and 16% in stage IIIB. The histological subtypes were: 32% squamous cell carcinoma, 50% adenocarcinoma, 14% anaplastic large cell, and 4% undifferentiated. 41% of the patients received second line chemotherapy. 1% achieved complete response (CR), 36% partial response (PR), 35% had stable disease (SD) and 28% progressive disease (PD). Patient’s median plasma levels of VEGF (20 pg/ml, [6–203]) differ significantly (p = 0.04) from controls (14 pg/ml, [7–53]), but in contrast bFGF levels were not different, 14 pg/ml [5–960] vs 10 pg/ml [6–278] respectively. There were not differences in patients according to histology, site of metastasis and ECOG; however we could observe a tendency with stage for both factors: bFGF 9 pg/ml [5–24] in stage IIIB vs 15 pg/ml [6–960], p = 0.071 and VEGF 17 pg/ml [6–145] in IIIB vs 21 pg/ml [6–203] in IV, p = 0.086. It could not be observed any differences in response to therapy for both angiogenic factors; CR+PR patients presented median VEGF of 18 pg/ml [6–71] and bFGF 11 pg/ml [6–960] vs 20 pg/ml of VEGF [6–203] and 15 pg/ml of bFGF [5–395] in the SD+PD group. In the multivariate analysis we could not find that VEGF and bFGF plasma levels were predictors for time to progression (TTP) and overall survival (OS). Conclusions: VEGF but not bFGF levels in patients are significantly higher in patients than in controls. In our cohort of patients with advanced NSCLC we have not found any relationship between serum VEGF and bFGF levels with stage, histology, response, site of metastasis, TTP and OS. No significant financial relationships to disclose.

Angiogenic Profile of Soft Tissue Sarcomas Based on Analysis of Circulating Factors and Microarray Gene Expression

Broader understanding of diverse angiogenic pathways in a particular cancer can lead to better utilization of anti-angiogenic therapies. The aim of this study was to develop profiles of angiogenesis-related gene and protein expression for various histologic subtypes of soft tissue sarcomas (STS) growing in different sites. Plasma levels of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), angiopoietin 2 (Ang2), and leptin were determined in 108 patients with primary STS. Gene expression patterns were analyzed in 38 STS samples and 13 normal tissues using oligonucleotide microarrays. VEGF and bFGF plasma levels were elevated 10-13 fold in STS patients compared to controls. VEGF levels were broadly elevated while bFGF levels were higher in patients with fibrosarcomas and leiomyosarcomas. Ang2 levels correlated with tumor size and were most elevated for tumors located in the trunk, while leptin levels were highest in patients with liposarcomas. Hierarchical clustering of microarray data based on angiogenesis-related gene expression demonstrated that histologic subtypes of STS often shared similar expression patterns, and these patterns were distinctly different from those of normal tissues. Matrix metalloproteinase 2, platelet-derived growth factor receptor, alpha and Notch 4 were among several genes that were up-regulated at least 7-fold in STS. STS demonstrate significant heterogeneity in their angiogenic profiles based on size, histologic subtype, and location of tumor growth, which may have implications for anti-angiogenic strategies. Comparison of STS to normal tissues reveals a panel of upregulated genes that may be targets for future therapies.

Elevated plasma levels of vascular endothelial growth factor is associated with marked splenomegaly in chronic myeloid leukemia

Recent investigations support the idea that angiogenesis is involved in the pathophysiology of hematologic malignancies, including chronic myeloid leukemia (CML). The aim of the present study was to evaluate plasma levels of VEGF and bFGF in a cohort of 51 chronic-phase CML patients at the time of diagnosis, as well as to investigate the effect of imatinib therapy on VEGF amounts in CML patients. Plasma VEGF levels were significantly higher in patients studied as compared with the 20 healthy subjects (p<0.001), the median plasma VEGF level detected in patients analysed and healthy controls was 433.4 pg mL−1 (range 65.2–2452.7 pg mL−1) and 81.6 pg mL−1 (range 44.2–338.7 pg mL−1), respectively. On the other hand, no difference in bFGF plasma levels could be found between chronic-phase CML patients and the control group. There were significant associations between plasma VEGF levels and some characteristics of patients evaluated, with trends for higher VEGF values in patients with enlarged spleens (p = 0.02) and those with higher platelet count (p<0.001). Of the patients, 11 received imatinib treatment. The initial VEGF levels markedly decreased after 6 months of imatinib therapy in each patient (p<0.001). These data support the important pathophysiological role of VEGF in CML. Further studies aiming to explore the detailed angiogenic profile of CML may help in developing new therapeutic strategies for this myeloproliferative disorder.

Elevated Basic Fibroblast Growth Factor Levels in Patients With Pulmonary Arterial Hypertension

Cellular growth in the vascular wall, including endothelial and smooth-muscle cell proliferation, is recognized as a component of the obstructive vasculopathy observed in the small vessels of the lungs in pulmonary arterial hypertension (PAH). We hypothesized that angiogenic growth factors may have a role in the molecular mechanisms underlying this cellular proliferation. Case-control study. Multicenter, tertiary care hospitals. We studied 117 patients with PAH and 60 control subjects. We measured levels of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) in the blood and urine of these subjects using an enzyme-linked immunoassay. Median levels of urinary and plasma bFGF were significantly higher in patients with PAH compared to normal control subjects. There was a difference in levels of urine and plasma bFGF according to etiology of pulmonary hypertension, with the highest levels seen in patients with primary pulmonary hypertension. Levels of urine or plasma VEGF were not significantly different between patients and control subjects. Patients with PAH have substantial alterations in urine and plasma levels of bFGF. This molecule may have a role as a mitogenic factor in the endothelial and smooth-muscle cell proliferation seen in PAH.

Bone marrow expression and plasma concentration of basic fibroblast growth factor in patients with idiopathic thrombocytopenic purpura.

Basic fibroblast growth factor (bFGF) is one of main regulators of hematopoiesis, including megakaryopoiesis. The main bone marrow finding of patients with idiopathic thrombocytopenic purpura (ITP) is increased megakaryopoiesis. The purposes of this study were to evaluate the change in the production of bFGF and its expression pattern in the bone marrow of patients with ITP and to correlate these characteristics with the plasma concentration of bFGF. Paraffin sections of bone marrow biopsies from 17 patients with ITP and 5 healthy control subjects without pathologic alterations were investigated by immunohistochemistry for bFGF and CD68. bFGF messenger RNA (mRNA) in situ hybridization was performed with bone marrow biopsy sections, and the plasma levels of bFGF were evaluated by enzyme immunoassay. bFGF was expressed strongly in stromal cells and weakly in megakaryocytes. The density of bFGF-expressing stromal cells was decreased in 70% (12/17) of the patients with ITP but in none of the control subjects. The numbers of stromal cells and the cellular distributions of bFGF mRNA in patients with ITP were similar to those of the control subjects. The plasma levels of bFGF were significantly lower in almost all of the ITP patients relative to those of the control group. The results indicate that the concentrations of bFGF in plasma and bone marrow stromal cells in ITP cases are decreased, whereas the production of bFGF remains unchanged. Although the mechanism of low cellular and plasma concentrations of bFGF needs to be elucidated, these findings may complement the serologic and morphologic diagnosis of ITP.

Exercise increases endostatin in circulation of healthy volunteers.

Physical inactivity increases the risk of atherosclerosis. However, the molecular mechanisms of this relation are poorly understood. A recent report indicates that endostatin, an endogenous angiostatic factor, inhibits the progression of atherosclerosis, and suggests that reducing intimal and atherosclerotic plaque tissue neovascularization can inhibit the progression atherosclerosis in animal models. We hypothesize that exercise can elevate the circulatory endostatin level. Hence, exercise can protect against one of the mechanisms of atherosclerosis. We examined treadmill exercise tests in healthy volunteers to determine the effect of exercise on plasma levels of endostatin and other angiogenic regulators. Oxygen consumption (VO2) was calculated. Plasma levels of endostatin, vascular endothelial growth factor (VEGF), and basic fibroblast growth factor (bFGF) were determined using ELISA. The total peak VO2 (L) in 7 male subjects was 29.5 +/- 17.8 over a 4-10 minute interval of exercise. Basal plasma levels of endostatin (immediately before exercise) were 20.3 +/- 3.2 pg/ml, the plasma levels increased to 29.3 +/- 4.2, 35.2 +/- 1.8, and 27.1 +/- 2.2 ng/ml, at 0.5, 2, and 6 h, respectively, after exercise. There was a strong linear correlation between increased plasma levels of endostatin (%) and the total peak VO2 (L) related to exercise (R2 = 0.9388; P < 0.01). Concurrently, VEGF levels decreased to 28.3 +/- 6.4, 17.6 +/- 2.4, and 26.5 +/- 12.5 pg/ml, at 0.5, 2, and 6 h, respectively, after exercise. There were no significant changes in plasma bFGF levels in those subjects before and after exercise. The results suggest that circulating endostatin can be significantly increased by exercise in proportion to the peak oxygen consumption under physiological conditions in healthy volunteers. These findings may provide new insights into the molecular links between physical inactivity and the risk of angiogenesis dependent diseases such as atherosclerosis.

The effect of a levonorgestrel-releasing intrauterine system on angiogenic growth factors in the endometrium.

The study aim was to investigate the effects of a levonorgestrel-releasing intrauterine system (LNG-IUS) on the secretion profiles and level of mRNA transcripts for angiogenic growth factors including vascular endothelial growth factor (VEGF), transforming growth factor (TGF)-beta1, basic fibroblast growth factor (bFGF) and endothelial growth factor (EGF) in the endometrium, and their levels in plasma. Blood and endometrial samples were collected from 15 women using LNG-IUS at pre-insertion and at 1, 3, 6 and 12 months following the use of contraception. Using enzyme-linked immunosorbent assay (ELISA) and semi-quantitative reverse transcription techniques, angiogenic growth factors were measured in the plasma and endometrial tissue extracts. A significant increase in endometrial tissue levels of VEGF at 3 and 6 months (P < 0.007, P < 0.01), TGF-beta1 at 6 and 12 months (P < 0.002, P < 0.007) and bFGF at 1 and 6 months (P < 0.004, P < 0.008) respectively were observed following insertion of LNG-IUS. A corresponding increase in mRNA expression of TGF-beta1 and bFGF were also observed. The LNG-IUS had no effect on endometrial EGF levels. No significant differences were noted in plasma levels of bFGF, EGF and TGF-beta1, while VEGF was not detectable in all plasma samples assayed. A positive correlation was observed between endometrial tissue VEGF and bleeding/spotting days in the reference period (r = 0.705, P < 0.015). These results provide the first evidence that more than one angiogenic factor could be implicated in aberrant endometrial angiogenesis resulting in breakthrough bleeding in LNG-IUS acceptors.

Selected Growth Factors in Peritoneal Dialysis: Their Relationship to Markers of Inflammation, Dialysis Adequacy, Residual Renal Function, and Peritoneal Membrane Transport

Markers of chronic inflammation, acute-phase reactants, and growth factors may be concomitantly involved in a number of pathologic processes in the general population and uremic patients. In addition, growth factors may influence peritoneal membrane transport characteristics. However, the association between plasma growth factors, markers of chronic inflammation, and peritoneal membrane transport remains largely unknown. The aim of this study was to evaluate the relationship between plasma levels of selected growth factors [basic fibroblast growth factor (bFGF), transforming growth factor beta1 (TGFbeta1), vascular endothelial growth factor (VEGF)] and markers of chronic inflammation [interleukin (IL)-6, C-reactive protein (CRP), and fibrinogen] in continuous ambulatory peritoneal dialysis (CAPD) patients. The potential link between the above substances and dialysis adequacy was also explored. Single-center, cross-sectional study. Peritoneal Dialysis Unit, Medical Faculty, Jagiellonian University Hospital, Kraków, Poland. 32 stable end-stage renal disease patients (13 M, 19 F; mean age 53.6 +/- 13.7 years) on CAPD for a median period of 19.5 months. Patients free from signs and symptoms of any inflammatory disease (including peritonitis) for at least 3 months were included into the study. All patients underwent measurements of dialysis dose [Kt/V, weekly creatinine clearance (wCCr)] and peritoneal solute transport using a standard peritoneal equilibration test (PET). TGFbeta1, bFGF, VEGF, and IL-6 were measured with ELISA, CRP was assayed with immunonephelometry, and fibrinogen with Multifibren U reagent (Dade Behring Marburg GmbH, Marburg, Germany). Nephron 97 for Windows software was used to assess dialysis adequacy. Significant positive correlations between plasma bFGF and IL-6, as well as fibrinogen concentrations (R = 0.36, p < 0.05 and R = 0.39, p < 0.05, respectively), were found. VEGF correlated significantly with IL-6 and CRP (R = 0.65, p < 0.0001 and R = 0.51, p < 0.005, respectively). An association between VEGF and bFGF was also found (R = 0.59, p < 0.0005). Serum level of TGFbeta1 revealed no relationship with any marker of acute-phase activation, remaining growth factors, or dialysis adequacy. Positive correlation between TGFbeta1 concentration and dialysate-to-plasma ratio for creatinine in PET (R = 0.35, p < 0.05) was found. In addition, patients with lower solute transport (low/low-average transporters) had lower serum levels of both bFGF and TGFbeta1 compared to patients with higher solute transport. Patients with total wCCr > 60 L/ week/m2 were characterized by lower levels of bFGF and IL-6. Serum level of IL-6 and plasma levels of bFGF and VEGF were significantly lower among subjects with residual renal function (RRF) > 2.0 mL/minute. Our results indicate that systemic inflammation in peritoneal dialysis patients is associated with increased plasma VEGF and bFGF but not TGFbeta1. The negative correlation with RRF suggests that either the renal clearance of these cytokines and growth factors may contribute to their elimination, or cytokines and growth factors have a negative impact on RRF. We also suggest an association between serum levels of growth factors tested and peritoneal membrane function.