Low Vascular Endothelial Growth Factor Expression Research Articles

Identifying the association of contrast enhancement with vascular endothelia growth factor expression in anaplastic gliomas: a volumetric magnetic resonance imaging analysis.

Contrast enhancement is a crucial radiologic feature of malignant brain tumors, which are associated with genetic changes of the tumor. The purpose of the current study was to investigate the potential relationship among tumor contrast enhancement with MR imaging, vascular endothelial growth factor (VEGF) expression, and survival outcome in anaplastic gliomas. MR images from 240 patients with histologically confirmed anaplastic gliomas were retrospectively analyzed. The volumes of T2 hyperintense, contrast enhanced regions and necrotic regions on postcontrast T1-weighted images were measured. The ratio of the enhanced volume to necrotic volume was compared between patients with high versus low levels of VEGF expression and was further used in the survival analysis. The volumetric ratio of enhancement to necrosis was significantly higher in patients with low VEGF expression than in those with high VEGF expression (Mann-Whitney, p = 0.009). In addition, the enhancement/necrosis ratio was identified as a significant predictor of progression-free survival (Cox regression model, p = 0.004) and overall survival (Cox regression model, p = 0.006) in the multivariate analysis. These results suggest that the volumetric ratio of enhancement to necrosis could serve as a noninvasive radiographic marker associated with VEGF expression and that this ratio is an independent predictor for progression-free survival and overall survival in patients with anaplastic gliomas.

Combined iNO and endothelial progenitor cells improve lung alveolar and vascular structure in neonatal rats exposed to prolonged hyperoxia.

Stem cells or inhaled nitric oxide (iNO) are reported to improve lung structures in bronchopulmonary dysplasia (BPD) models. We hypothesized that combined iNO and transplanted endothelial progenitor cells (EPCs) might restore lung structure in rats after neonatal hyperoxia. Litters were separated into eight groups: room air, hyperoxia, hyperoxia + iNO, hyperoxia + iNO + L-NAME, hyperoxia + EPCs, hyperoxia + EPCs + L-NAME, hyperoxia + EPCs + iNO, and hyperoxia + EPCs + iNO + L-NAME. Litters were exposed to hyperoxia from the 21st day, then, sacrificed. EPCs were injected on the 21st day. L-NAME was injected daily for 7 d from the 21st day. Serum vascular endothelial growth factor (VEGF), radial alveolar count (RAC), VIII factor, EPCs engraftment, lung VEGF, VEGFR2, endothelial nitric oxide (eNOS) and SDF-1 expression, and NO production were examined. Hyperoxia exposure led to air space enlargement, loss of lung capillaries, and low expression of VEGF and eNOS. Transplanted EPCs, when combined with iNO, had significantly increased engraftment in lungs, compared to EPCs alone, upon hyperoxia exposure. There was improvement in alveolarization, microvessel density, and upregulation of VEGF and eNOS proteins in the hyperoxia-exposed EPCs with iNO group, compared to hyperoxia alone. Combined EPCs and iNO improved lung structures after neonatal hyperoxia. This was associated with the upregulation of VEGF and eNOS expression.

Serum VEGF Predicts Worse Clinical Outcome of Patients with Coronary Heart Disease After Percutaneous Coronary Intervention Therapy.

BackgroundPercutaneous coronary intervention (PCI) is an effective treatment for coronary heart disease (CHD) patients. However, patients after PCI treatment often have ischemic events that result in poor prognosis. Our study aimed to investigate the effects of vascular endothelial growth factor (VEGF) level on the prognosis of CHD patients.Material/MethodsWe enrolled 114 CHD patients in the study. Serum VEGF level was measured by enzyme-linked immunosorbent assay (ELISA). Total cholesterol, LDL cholesterol, HDL cholesterol, triglycerides, and Hs-CRP were also tested in patients. The patients were divided into 2 groups according to the level of VEGF. Kaplan-Meier curve was used to observe the differences in survival situation of patients of the 2 groups. Cox regression analysis was conducted to judge whether VEGF was an independent biomarker for prognosis in CHD.ResultsWe included 104 patients for survival analysis. VEGF level in CHD patients was significantly lower than that of healthy individuals (P<0.05). In the analysis of basic information, we found differences in sex distribution and hypertension between groups (P<0.05 for both). Kaplan-Meier curve indicated that patients with low expression of VEGF presented with poor prognosis. The mortality rate of the low-expression group was 37.71%, higher than that of the high-expression group (14.3%). Cox analysis suggested that VEGF could serve as a biomarker for prognosis in CHD (HR: 3.014, P: 0.019).ConclusionsLow level of VEGF may predict poor clinical outcome of CHD patients after PCI treatment.

Correlation between the expression of vegf and survival in osteosarcoma

Objective: To present a series of 50 consecutive patients with non-metastatic extremity osteosarcoma, and attempt to correlate expression of the vascular endothelial growth factor (VEGF) protein in biopsy tissue to their prognosis regarding overall survival, disease-free survival and local recurrence. Methods: Fifty cases of non-metastatic osteosarcoma of the extremities treated between 1986 and 2006 at Instituto de Ortopedia e Traumatologia da Universidade de São Paulo, São Paulo, Brasil, were evaluated regarding expression of the VEGF protein. There were 19 females and 31 males. The mean age was 16 years old (range 5-28 years old) and the mean follow-up was 60.6 months (range 25-167 months). The variables studied were age, gender, anatomic location, type of surgery, surgical margins, tumor size, post chemotherapy necrosis, local recurrence, pulmonary metastasis and death. Results: Thirty-six patients showed VEGF expression on 30% or less cells (low), and the remaining 14 cases had VEGF expression above 30% (high). Among the 36 patients with low VEGF expression, nine developed pulmonary metastasis and four died (11.1%). Among the 14 patients with high VEGF expression, six developed pulmonary metastasis and three died (21.4%). Conclusion: There was no statistically significant correlation between the expression of VEGF and any of the variables studied. Level of Evidence IV, Therapeutic Study.

Correlation between ¹⁸F-fluoromisonidazole PET and expression of HIF-1α and VEGF in newly diagnosed and recurrent malignant gliomas.

Hypoxia and its consequences at the molecular level promote tumour progression and affect patient prognosis. One of the main early cellular events evoked by hypoxia is induction of hypoxia-inducible factor 1 (HIF-1) and subsequent upregulation of vascular endothelial growth factor (VEGF). In this study we sought to determine whether hypoxia detected by (18)F-fluoromisonidazole (FMISO) PET accurately reflects the expression of HIF-1α and VEGF in the tumour and can be used as a biomarker of antiangiogenic treatment and as a prognostic factor in newly diagnosed and recurrent malignant gliomas. Enrolled in this study were 32 patients with newly diagnosed glioma and 16 with recurrent glioma of grade III or grade IV. All the patients had undergone FMISO PET preoperatively. The maximum tumour-to-blood FMISO activity ratio (T/Bmax) was used to evaluate the degree of tumour hypoxia and the hypoxic volume (HV) was calculated using a tumour-to-blood FMISO uptake ratio of ≥1.2. Immunohistochemical expressions of HIF-1α and VEGF were evaluated semiquantitatively using the immunoreactivity score (IRS, scores 0 to 12) and the correlation was examined between IRS of HIF-1α or VEGF and FMISO uptake of the tumour (SUVtumour) using navigation-based sampling. Survival was estimated using the Kaplan-Meier method in relation to the T/Bmax and the HV. The T/Bmax and the HV in grade IV gliomas were significantly higher than in grade III gliomas (P < 0.01 and P < 0.01, respectively). Moderate to strong HIF-1α and VEGF expression was observed in the majority of malignant gliomas. The IRS of HIF-1α and VEGF in the tumour were not significantly different between grade III and grade IV gliomas. The IRS of HIF-1α in the tumour did not correlate with the SUVtumour of FMISO in either newly diagnosed or recurrent glioma. There was a significant but weak correlation between the IRS of VEGF and the SUVtumour of FMISO in newly diagnosed glioma, but not in recurrent glioma. The overall survival time in patients with a small HV and a low FMISO T/Bmax was significantly longer than in those with a large HV and a high FMISO T/Bmax (P < 0.01 and P < 0.05, respectively). Preoperative FMISO uptake is significantly correlated with the expression of VEGF in the tumour and might be used as a biomarker of antiangiogenic treatment in newly diagnosed malignant gliomas. However, caution is required because the correlation was weak and there was a large overlap of FMISO uptake between glioma with high and low VEGF expression. In addition, hypoxia determined by FMISO PET appears to be a suitable biomarker for predicting a highly malignant tumour and a poor prognosis in patients with malignant glioma.

Role of vascular endothelial growth factor in clinically localized prostate cancer treated with radiation therapy.

Anti-vascular endothelial growth factor (Anti-VEGF) agents are a promising approach to increase the efficacy of treatment for treatment-resistant prostate cancer. To correlate vascular endothelial growth factor (VEGF) expression and outcome following radiation therapy in the treatment of clinically localized prostate cancer. Retrospective observational study. Forty-one patients and clinically localized disease that were treated with radiation therapy were analyzed. For VEGF expression, immunoreactivity scores (IRS) were calculated using percent scores and intensity scores. Twenty-four patients were classified as having low (0 to 4 IRS) and 17 patients were classified as having high (5 to 8 IRS) VEGF expression. The median age was 71 years, median follow-up was 5.4 years and median radiation therapy dose was 70 Gy. VEGF expression was calculated as low in 24 patients and high in 17 patients. Higher VEGF expression was observed in 6/26 patients with a low Gleason score versus 11/15 patients with a high Gleason score (p=0.02). Biochemical failure (BF) was observed in 2/24 patients with low VEGF expression versus 7/17 patients with high VEGF expression (p=0.01). In univariate analysis, having a higher Gleason score (p<0.01), being in the high risk group (p=0.03) and having higher VEGF expression (p=0.01) predicted BF after definitive radiation therapy. The biochemical failure-free survival rate at 5 years tended to be different (91% vs. 53%) when patients were grouped according to VEGF expression (p=0.06). In attempt to define patients with clinically localized disease that are not sensitive to standard treatment modalities, cellular and/or molecular biological markers may be required.

Protective effect of lycopene against diabetic retinopathy in rats and the related mechanism

Objective To study the protective effect of lycopene(LP)against diabetic retinopathy(DR)in rats and its related mechanism.Methods A single dose of streptozotocin(56mg/kg)was injected intraperitoneally in SD rats to induce diabetes retinopathy(DR),and the 20 successful DR models were equally randomized into DR group and LP treatment group.Meanwhile,10 normal rats served as normal control group(NC).LP group received LP gavage(60mg/kg)once a day;NC and DR groups were gavaged with equivalent saline daily for 8weeks.The serum levels of endothelin(ET)and calcitonin gene related protein(CGRP)were detected by using radioimmunoassay(RIA),and the expression of vascular endothelial growth factor(VEGF)in the retina was determined by immunohistochemistry and Western blotting analysis.Results The serum levels of ET and expression of VEGF in the retina were significantly increased in DR groups when compared with NC group(P0.05),and the serum CGRP was significantly decreased(P0.05);Compared with DR group,LP group had significantly lower VEGF expression and ET levels and significantly higher level of CGRP(P0.05).Conclusion LP can protect against DR in rats by decreasing serum ET,increasing serum CGRP,and subsequent down-regulating VEGF expression.

Role of Activated Rac1/Cdc42 in Mediating Endothelial Cell Proliferation and Tumor Angiogenesis in Breast Cancer

Angiogenesis is a well-established target in anti-cancer therapy. Although vascular endothelial growth factor (VEGF)-mediated angiogenesis apparently requires the Rho GTPases Rac1 and Cdc42, the relevant mechanisms are unclear. Here, we determined that activated Rac1/Cdc42 in MCF-7 breast cancer cells could decrease p53 protein levels and increase VEGF secretion to promote proliferation and tube formation of human umbilical vein endothelial cells (HUVECs). However, these effects are reversed after ubiquitin-proteasome breakage. In exploring potential mechanisms for this relationship, we confirmed that activated Rac1/Cdc42 could enhance p53 protein ubiquitination and weaken p53 protein stability to increase VEGF expression. Furthermore, in a xenograft model using nude mice that stably express active Rac1/Cdc42 protein, active Rac1/Cdc42 decreased p53 levels and increased VEGF expression. Additionally, tumor angiogenesis was inhibited, and p53 protein levels were augmented, by intratumoral injection of the ubiquitin-proteasome inhibitor MG132. Finally in 339 human breast cancer tissues, our analyses indicated that Rac1/Cdc42 expression was related to advanced TNM staging, high proliferation index, ER status, and positive invasive features. In particular, our data suggests that high Rac1/Cdc42 expression is correlated with low wt-p53 and high VEGF expression. We conclude that activated Rac1/Cdc42 is a vascular regulator of tumor angiogenesis and that it may reduce stability of the p53 protein to promote VEGF expression by enhancing p53 protein ubiquitin.

A systematic review of vascular endothelial growth factor expression as a biomarker of prognosis in patients with osteosarcoma

Vascular endothelial growth factor (VEGF) plays an important role in the tumor angiogenesis, and its expression has been supposed to be a biomarker of prognosis in patients with osteosarcoma. There are many studies assessing the prognostic role of VEGF expression in osteosarcoma, and no consistent outcomes are reported. To provide a comprehensive assessment of the prognostic role of VEGF expression, we performed a systematic review and meta-analysis of published studies. We assessed the effect of VEGF expression on the overall survival rate and the disease-free survival rate by calculating the pooled odds ratio (OR) with corresponding 95 % confidence interval (95 %CI). Finally, 12 studies with a total of 559 osteosarcoma patients were included into the systematic review and meta-analysis. Compared with osteosarcoma patients with low or negative VEGF expression, patients with high VEGF expression were obviously associated with lower disease-free survival (OR=0.25, 95 %CI 0.11-0.58, P=0.001, I (2) =56.4 %). In addition, patients with high VEGF expression were obviously associated with lower overall survival (OR=0.22, 95 %CI 0.13-0.35, P<0.001, I (2) =0.0 %). Therefore, the findings from this systematic review suggest that VEGF expression is an effective biomarker of prognosis in patients with osteosarcoma.

Expression of vascular endothelial growth factor (VEGF) and its receptors VEGFR1 and VEGFR2 in primary and recurrent WHO grade III meningiomas.

WHO grade III meningiomas are malignant neoplasms for which new and more targeted treatment strategies are urgently needed. Although clinical trials investigating anti-angiogenic vascular endothelial growth factor (VEGF) targeted therapies are currently recruiting, knowledge about the expression of VEGF and VEGF receptors remains to be determined. We investigated the expression of VEGF and its receptors VEGFR1 and VEGFR2 in 32 WHO grade III meningioma samples by immunohistochemistry. Furthermore, we performed in-situ hybridisation for VEGF. We found low VEGF expression in tumor and endothelial cells. Highest VEGF expression levels were seen in peri-necrotic tumor cells potentially suffering from hypoxia. VEGFR1 and 2 were virtually absent on tumor cells, although endothelial cells displayed significantly higher levels reaching stronger expression for VEGFR2 than VEGFR1. Our findings showing constant expression levels of VEGFR2 in endothelial cells serve as a first indication that the use of small tyrosine kinase inhibitors such as Sunitinib directly targeting the VEGF-receptors might be worth testing, also in the clinical context in cases of therapy-refractory meningiomas. Further investigations are needed to study the response to drugs targeting the VEGF pathway in relation to the expression profile of VEGF and its receptors in high grade meningiomas.

Necrosis Degree Displayed in Computed Tomography Images Correlated With Hypoxia and Angiogenesis in Breast Cancer

The objective of this study was to investigate the correlation between the degree of necrosis displayed in computed tomography (CT) image and the expression of hypoxic and angiogenesis biomarkers of breast cancer. Forty-four breast cancer cases were examined with CT before surgery. Tumor specimen expressions of glucose transporter 1 (Glut1), hypoxia-inducible factor 1α (HIF-1α), carbonic anhydrase IX (CA IX), vascular endothelial growth factor (VEGF), and CD34 (as a marker of vascular endothelial cells) were detected by immunohistochemistry. The expressions of Glut1 and CA IX were localized primarily to the edges of necrotic areas or intraduct surface; there was a strong correlation between HIF-1α-positive expression and CA IX-positive expression (P < 0.001), and higher Glut1 or CA IX expression grade was associated with lower microvessel density (MVD) value. In CT enhanced images, lower relative CT (rCT) values were associated with more significant necrosis in the tumor; rCT value correlated positively with MVD significantly (r = 0.319, P = 0.035), and higher Glut1 or CA IX expression grade correlated with lower rCT values (P = 0.001 and 0.003, respectively). Although high VEGF expression was significantly correlated with high HIF-1α expression (P < 0.001), there were no correlations between VEGF and MVD, and HIF-1α and MVD (P = 0.559 and 0.710, respectively), and the difference of the mean rCT value between high VEGF or HIF-1α expression group and low VEGF or HIF-1α expression group was not significant. In breast cancer tissues, Glut1 and CA IX are key hypoxia biomarkers. Importantly, detection of necrosis in breast cancer tissue via CT enhanced imaging may prognosticate hypoxia and angiogenesis status and help to determine treatment plan of advanced breast cancer.

First In Vivo Application of Microwave Radiometry in Human Carotids: A New Noninvasive Method for Detection of Local Inflammatory Activation

This study investigated whether temperature differences: 1) can be measured in vivo noninvasively by microwave radiometry (MR); and 2) are associated with ultrasound and histological findings. Studies of human carotid artery samples showed increased heat production. MR allows in vivo noninvasive measurement of internal temperature of tissues. Thirty-four patients undergoing carotid endarterectomy underwent screening of carotid atherosclerosis by ultrasound and MR. Healthy volunteers were enrolled as a control group. During ultrasound study, plaque texture, plaque surface, and plaque echogenicity were analyzed. Temperature difference (ΔT) was assigned as maximal minus minimum temperature. Association of thermographic with ultrasound and histological findings was performed. ΔT was higher in atherosclerotic carotid arteries compared with the carotid arteries of controls (p < 0.01). Fatty plaques had higher ΔT compared with mixed and calcified (p < 0.01) plaques. Plaques with ulcerated surface had higher ΔT compared with plaques with irregular and regular surface (p < 0.01). Heterogeneous plaques had higher ΔT compared with homogenous (p < 0.01). Specimens with thin fibrous cap and intense expression of CD3, CD68, and vascular endothelial growth factor (VEGF) had higher ΔT compared with specimens with thick cap and low expression of CD3, CD68, and VEGF (p < 0.01). MR provides in vivo noninvasive temperature measurements of carotid plaques, reflecting plaque inflammatory activation.

Abstract 5278: Suppression of angiogenic potential of breast cancer by the inhibition of SAF-1 transcription factor activity

Abstract Angiogenesis plays an important role in the tumor growth and metastasis by facilitating increased blood supply and thus fueling unwanted growth factors and nutrients to the growing tumor. Vascular endothelial growth factor (VEGF) is recognized as a multifunctional angiogenic stimulator that stimulates blood vessel formation and endothelial cell survival. Recent therapeutic trials with anti-VEGF antibody bevacizumab (Avastin), although produced beneficial effects on some cancers including colorectal and lung cancers, had very limited beneficial outcome in breast cancer. The diverse response of avastin raised questions about the efficacy of anti-VEGF antibody approach for breast cancer therapy. Despite the disappointing outcome, it cannot be ignored that regulation of angiogenesis plays a critical role in breast cancer. We hypothesized that angiogenesis may be better controlled by blocking the synthesis of VEGF protein as opposed to the present approach that attempts to block the functions of VEGF by using antibodies. Induction of VEGF by a plethora of physiological and environmental stimuli strongly suggests the presence of multiple routes of VEGF synthesis, which are incompletely understood. We present evidence for a novel mode of transcriptional induction of VEGF, regulated by an inflammation-responsive transcription factor, SAF-1. Incidentally, inflammatory breast cancer exhibits increased angiogenesis and has a higher metastatic potential than noninflammatory breast cancer, supporting the notion that chronic inflammation plays a strong role in angiogenesis and in the progression of cancer. SAF-1 is present in high abundance in human breast cancer tissues. We show that inhibition of endogenous SAF-1 by antisense shRNA can markedly inhibit VEGF expression and block cancer cell-supported vascular endothelial cell function and angiogenesis. Furthermore, SAF-1 inhibition by shRNA reduces tumor growth in mouse tumor model. We provide evidence that SAF-1-mediated induction of VEGF remains suppressed by KLF-4 transcription factor which is abundantly present in normal breast epithelial cells but absent in breast cancer cells. Mutually exclusive interaction of KLF-4 and SAF-1 at the VEGF promoter provides a mechanism for regulation of low-expression of VEGF in normal breast tissues. In correlation, lack of KLF-4 in breast cancer cells appears to be linked to un-restricted interaction of SAF-1 leading to the induction of VEGF expression. Together, our studies provide a new insight into the molecular mechanisms of VEGF expression regulated by SAF-1 and its impact in angiogenesis. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5278. doi:1538-7445.AM2012-5278

Low VEGF expression in conceptus material and maternal serum AFP and levels as indicators of defective angiogenesis in first-trimester miscarriages

The aims of this study were to assess the relationship between early miscarriages and vascular endothelial growth factor (VEGF) expression and to determine the serum levels of first-trimester maternal alpha-fetoprotein (AFP) and human chorionic gonadotropin (β-hCG) as markers of angiogenesis and predictors of abortion and intrauterine fetal loss. The present study was a prospective, single-center, randomized controlled clinical trial. Ninety-five women who were 6-10 weeks pregnant between May and June 2010 were included in the study. The subjects were divided into three groups, i.e., incomplete abortion (IA) (n=31), intrauterine death (IU-D) (n=32) and control (elective pregnancy termination) (n=32). Feto-placental materials were compared based on immune staining for VEGF in the pathology laboratory, and maternal serum samples were tested in the hormone laboratory. Serum β-hCG levels in the patient groups were significantly lower than the controls (p=0.001). The serum AFP level was lower than the controls in the IA group while it was higher than the controls in the IU-D (p=0.016). Immunohistochemistry showed that the cytotrophoblast, syncytiotrophoblast and endometrial gland epithelium were weakly stained for VEGF in the patient groups (IA and IU-D) in comparison to the control group (p=0.06, p=0.028, p=0.006). Early pregnancy losses are related to insufficient angiogenesis, and maternal serum AFP and β-hCG can be used as markers of angiogenesis in the first trimester.

Abstract C56: Characterization of hz515H7, a novel humanized anti-CXCR4 antibody: In vitro efficacy on CXCR4-associated signaling pathways and in vivo antitumor activity.

Abstract C56: Characterization of hz515H7, a novel humanized anti-CXCR4 antibody: In vitro efficacy on CXCR4-associated signaling pathways and in vivo antitumor activity.

The inflammatory stimulus of a natural latex biomembrane improves healing in mice

The aim of the present study was to compare healing obtained with biomembranes with the natural healing process (sham) using biochemical and immunohistological assays. C57BL/6 mice were divided into 4 groups of 15 mice each and received different subcutaneous implants: natural latex biomembrane (NLB), denatured latex (DL), expanded polytetrafluorethylene (ePTFE), or sham. On the 2nd, 7th, and 14th days post-treatment, 5 mice per group were sacrificed and biopsied for the following measurements: oxidative stress based on malondialdehyde (MDA), myeloperoxidase (MPO) and hydrogen peroxide by the method of ferrous oxidation-xylenol orange (FOX), as well as glutathione and total proteins; histological evaluation to enumerate inflammatory cells, fibroblasts, blood vessels, and collagen, and immunohistochemical staining for inducible nitric oxide synthase, interleukin-1β, vascular endothelial growth factor (VEGF), and transforming growth factor-β1 (TGF-β1). On day 2 post-treatment, NLB stimulated a dense inflammatory infiltrate mainly consisting of polymorphonuclear cells, as indicated by increased MPO (P < 0.05), but oxidative stress due to MDA was not observed until the 7th day (P < 0.05). The number of blood vessels was greater in NLB (P < 0.05) and DL (P < 0.05) mice compared to sham animals on day 14. NLB induced fibroplasia by day 14 (P < 0.05) with low expression of TGF-β1 and collagenesis. Thus, NLB significantly induced the inflammatory phase of healing mediated by oxidative stress, which appeared to influence the subsequent phases such as angiogenesis (with low expression of VEGF) and fibroplasia (independent of TGF-β1) without influencing collagenesis.

Effects of polydeoxyribonucleotide on the histological damage and the altered spermatogenesis induced by testicular ischaemia and reperfusion in rats

The effects of polydeoxyribonucleotide (PDRN), an agonist of the A2A adenosine receptors which when activated positively influences sperm activity, were tested in an experimental testicular ischaemia/reperfusion injury model. Anaesthetized male Sprague-Dawley rats were subjected to testicular torsion-induced ischaemia, followed by reperfusion (TI/R). Immediately after detorsion, randomized animals, including SHAM, received intraperitoneal injections of: (i) vehicle (1 mL/kg 0.9% NaCl solution); (ii) PDRN (8 mg/kg); (iii) DMPX (3,7-dimethyl-1-propargilxanthine, 0.1 mg/kg); or (iv) PDRN (8 mg/kg) + DMPX (0.1 mg/kg). Animals were euthanized at 1, 7 and 30 days following reperfusion. Vascular endothelial growth factor (VEGF) expression is normally associated with adenosine A2A receptor stimulation. After treatment, VEGF mRNA/protein expression quantified by qPCR and Western blot, vascular endothelial growth factor receptor-1 (VEGFR1) and endothelial nitric oxide synthase (eNOS) mRNA measured by qPCR, VEGF and VEGFR1 assessed using immunohistochemical methods, histological staining and spermatogenic activity were all analysed. Testis ischaemia-reperfusion (TI/R) injury caused increases in VEGF mRNA and protein, VEGFR1 and eNOS mRNA, histological damage and reduced spermatogenic activity. Immunostaining showed a lower expression of VEGF in germinal epithelial cells and a strong expression of VEGFR1 in Leydig cells after TI/R. PDRN administration increased significantly VEGF message/protein, VEGFR1 and eNOS message, decreased histological damage and ameliorated spermatogenic activity. PDRN might be useful in the management of testicular torsion.

Vascular Endothelial Growth Factor and Its Receptors Are Expressed in Human Nerve Grafts

Vascularization and angiogenicity of human nonvascularized nerve grafts in the second stage of facial reanimation were studied. Immunohistochemistry for endothelial markers (CD-31) and vascular endothelial growth factor (VEGF) and its receptors Flt-1 and Flk-1 was performed on distal end biopsies from 35 cross-facial nerve grafts. In grafted nonvascularized nerve, density of vascular structures (also clearly immunopositive for VEGF and both receptors) showed a mean of 166 vessels (range 78 to 267) per unit area, corresponding to control values. In addition, VEGF was expressed in axons and perineural structures. In control samples, VEGF expression was low and occurred in the myelin sheath. In nerve grafts, expression of Flt-1 and Flk-1 (less intense) was seen in axons and perineural structures. A higher density of vessels was associated with lower VEGF expression (not significant). In short, expression of VEGF and its receptors is described in human nerve grafts and compared with basic histology and p75 nerve growth factor receptor expression of the nerve graft and functional outcome of patients.

Vascular endothelial growth factor and vascular endothelial growth factor receptor‐2 tumor expression in patients with advanced laryngeal cancer after induction chemotherapy for organ preservation

Vascular endothelial growth factor (VEGF) expression seems to be associated with worse overall survival in patients with head and neck squamous cell carcinoma. The purposes of this study were to assess the prognostic values of the immunohistochemical expression of VEGF and vascular endothelial growth factor receptor-2 (VEGFR-2) in a cohort of patients with operable advanced laryngeal cancer who had been treated with induction chemotherapy. VEGF and VEGFR-2 expression in the malignant cells was quantified by an enzyme immunosorbent assay in a retrospective series. Fifty-nine patients were included in this study. We found moderate to high VEGF expression in 61% of patients and none to low expression in 39%. VEGFR-2 expression was moderate to high in 58% of patients and was none to low in 42%. The probability of a complete response to induction chemotherapy was significantly higher in patients with none to low VEGF expression (74% vs 40%; p = .02). VEGF expression seems to be a significant predictor of complete response to induction chemotherapy.

113. VASCULAR ENDOTHELIAL GROWTH FACTOR GENE POLYMORPHISMS IN PLACENTAL IMPAIRMENT AND SMALL FOR GESTATIONAL AGE BIRTH

Impaired placental angiogenesis is implicated in the pathophysiology of small for gestational age (SGA) infants. Placental expression of vascular endothelial growth factor (VEGF), a potent angiogenic factor, is reduced in SGA pregnancies. We aimed to evaluate the association of two single nucleotide polymorphisms (SNPs, VEGF-2578C/A and VEGF+936C/T) in VEGF gene which reduce VEGF expression, in SGA pregnancies andexamine their effects on first trimester placental VEGF expression. 3196 nulliparous pregnant women, their partners and babies were recruited in Adelaide and Auckland to a prospective multicentre cohort study (SCOPE Study). Amongst 2123 Caucasian women, 216 (11.9%) delivered a SGA infant defined as &lt;10th customised centile. Uncomplicated Caucasian pregnancies served as controls (n = 1176). Uterine and umbilical artery Doppler velocimetry was performed at 20 weeks gestation. DNA extracted from peripheral blood from couples and cord blood from babies was genotyped using Sequenom MassARRAY. 74 first trimester placentae collected from elective terminations of pregnancy were genotyped for the same SNPs and the VEGF expression determined by RT-PCR. Neonatal VEGF+936 CT+TT genotypes associate with SGA (OR 1.6, 95%CI 1.1–2.3), lower birthweight (P = 0.005), customised birthweight centile (p=0.03), lower placental weight (P = 0.04) and an increased uterine artery resistance index (RI, P = 0.004). Maternal VEGF+936 CT+TT associate with bilateral notching of the uterine artery waveform (OR 1.4, 95%CI 1.0–1.8) and an increased umbilical artery RI (OR 1.5, 95%CI 1.1–2.1). VEGF+936 CT first trimester placentae have lower VEGF expression compared to CC (P = 0.045). Neonatal VEGF-2578 AA associates with bilateral uterine artery notching (OR 1.5, 95%CI 1.1–2.2) and increased umbilical artery RI (OR 1.6, 95%CI 1.0–2.6). Maternal VEGF-2578 CA+AA associate with increased umbilical artery RI (OR 1.5, 95%CI 1.0–2.2). VEGF polymorphisms reduce first trimester VEGF expression and associate with increased resistance in the placental circulation suggesting impaired placental function. VEGF+936 SNP confers increased risk for SGA.