Vascular Endothelial Growth Factor R2 Research Articles

Assessment of VEGF and VEGF R1 serum levels in patients with neuroendocrine neoplasms before and after treatment with first-generation somatostatin analogues.

Vascular endothelial growth factor (VEGF) is a known promoter of angiogenesis that can support neuroendocrine neoplasm (NEN) development. The aim of the study was to evaluate the serum VEGF and vascular endothelial growth factor receptor 1 (VEGF R1) concentration changes in patients with NEN treated with first-generation long-acting somatostatin analogues (SSA). The study comprised 55 controls and 56 NEN patients before and after SSA treatment in various periods of time (months): 1-12 (n = 54), 13-24 (n = 46), 25-36 (n = 35), 37-60 (n = 26), and over 60 months (n = 22). An analysis of medical records and serum VEGF and VEGF R1 concentration measurements of NEN patients, by enzyme-linked immunosorbent assay (ELISA) were made. During SSA treatment time, a decrease of the VEGF and an increase of VEGF R1 concentrations was observed. We confirmed significant VEGF differences between 2 pairs of SSA-treated NEN patient subgroups: Group 1-12 vs. Group 37-60 (p = 0.039) and Group 1-12 vs. Group > 60 (p = 0.026). We did not note significant differences of VEGF R1 levels between SSA-treated NEN patient subgroups. Among the studied biomarkers, VEGF R1 exhibited the best performance in distinguishing between NEN patients with controls; area under the curve (AUC) = 1 (p < 0.001). The examined angiogenesis factors (VEGF and VEGF R1) seem to have limited usage in the assessment of SSA treatment effectiveness in NEN. However, the assessment of serum levels of these factors may help in the differentiation of NEN patients and healthy controls; in particular, VEGF R1 seems to be a good diagnostic biomarker for NEN patients.

In Vitro Effects of Papaverine on Cell Migration and Vascular Endothelial Growth Factor in Cancer Cell Lines.

Papaverine (PPV) is a benzylisoquinoline alkaloid isolated from Papaver somniferum that exerts antiproliferative activity. However, several questions remain regarding the biochemical pathways affected by PPV in tumourigenic cells. In this study, the influence of PPV on cell migration (light microscopy), expression of vascular endothelial growth factor (VEGF) B, VEGF R1, VEGF R2, and phosphorylated focal adhesion kinase (pFAK) were investigated using spectrophotometry in MDA-MB-231-, A549- and DU145 cell lines. The migration assay revealed that, after 48 h, PPV (100 µM) reduced cell migration to 81%, 91%, and 71% in MDA-MB-231-, A549-, and DU145 cells, respectively. VEGF B expression was reduced to 0.79-, 0.71-, and 0.73-fold after 48 h of exposure to PPV in MDA-MB-231-, A549- and DU145 cells, while PPV exposure of 48 h increased VEGF R1 expression in MDA-MB-231- and DU145 cells to 1.38 and 1.46. A fold decrease in VEGF R1 expression was observed in A549 cells to 0.90 after exposure to 150 µM. No statistically significant effects were observed on VEGF R2- and FAK expression after exposure to PPV. This study contributes to the understanding of the effects of a phytomedicinal alkaloid compound in cancer cells and may provide novel approaches to the application of non-addictive alkaloids.

Фетоплацентарный ангиогенез при преэклампсии. Клинико-морфологические аспекты

Purpose. To study placental angiogenesis in pregnant women with preeclampsia (PE), based on a comparison of the results of a clinical examination and a morphometric study of sequins with this pathology. Material and methods. In the present work, we studied 30 pregnant women with PE, whose average age was 29 ± 1.4 years. The pregnant women were divided into two groups. Group I (n = 21) consisted of pregnant women with PE, among whom 10 patients were within 34 weeks of gestation, and 11 – at 34–39 weeks. Group II (n = 9) – control group, with physiological pregnancy. The inclusion criteria were: singleton pregnancy, the presence of preeclampsia according to the ICD-10 classification, the patient's voluntary informed consent to the study. Exclusion criteria: extragenital pathology, multiple births, congenital malformations of the fetus. There were no significant differences in somatic and gynecological status among pregnant women in the surveyed groups. Results. Vascular endothelial growth factor (VEGF) and its receptors, VEGF R1 and VEGF R2, are of priority importance in the development of placental angiogenesis. The balance of interaction between VEGF and R1 and R2 receptors leads to adequate vascularization of the villi in the early stages of trophoblast invasion into the spiral arteries of the uterus. Conclusion. As a result of the morphological study, it can be assumed that hemodynamic parameters according to Doppler and angioarchitectonics of the placentas are an important factor in the pathogenesis of placental insufficiency and fetal growth retardation. In the absence of disturbances in uteroplacental-fetal hemodynamics, endothelial dysfunction should be considered as compensated, in which favorable perinatal outcomes are expected. Structural changes in the fetoplacental complex with impaired blood flow in the uterine arteries and umbilical cord arteries in PE are regarded as subcompensated and decompensated forms of placental insufficiency

Abstract 13040: Left Ventricle Dysfunction is Prevented by the Treatment With Methotrexate Carried in Lipid Nanoparticles in Lipopolysaccharide-injected Rats

Introduction: In infectious processes, left ventricle (LV) dysfunction is often associated with dysregulation of the immune response. LDE, a lipid core nanoparticle resembling LDL, is able to concentrate in inflammatory sites. We showed that methotrexate (MTX), associated with LDE, modulated inflammation and improved cardiac function in rats with myocardial infarction Hypothesis: Investigate therapeutic effects of LDE-MTX on LV function in lipopolysaccharide (LPS)-injected rats Methods: Wistar rats were submitted to 2 injections of LPS (10mg/kg ip), with 24h interval. Animals were allocated to control group (CT, without LPS) and 3 LPS groups: LDE, treated with only LDE; MTX, with commercial MTX; and LDE-MTX. LDE-MTX dose was 1mg/kg I.P. 72h after LPS, animals were submitted to echocardiographic analysis. Animals were euthanised for analysis of the LV Results: Rats treated with LDE developed diastolic dysfunction and presented decreased diastolic volume and diameter of the LV, which was prevented by LDE-MTX. Only LDE-MTX treatment elicited cardiac hypertrophy through the increase of the thickness of LV septum and posterior wall, LV mass, relative heart weight and myocyte diameter. Inflammatory and apoptotic markers were similar in LPS groups, but LDE-MTX treatment increased macrophage recruitment (CD68). Angiogenesis, represented by vascular endothelial growth factor (VEGF), was higher in LDE-MTX compared to all other groups. Hypoxia-inducible factor 1 alpha, which was remarkably higher in LDE, was normalized by LDE-MTX to CT levels. A1, A2b and A3 adenosine receptors expression was higher in LDE-MTX, compared to CT, indicating an increase in intracellular adenosine. Myocyte diameter was positively correlated with VEGF (r2= 0.52; p&lt;0.0001) Conclusions: The increased recruitment of immune cells to the LV indicates that treatment with LDE-MTX was effective in activating immune response, impaired by LPS. LDE-MTX increased intracellular adenosine availability, contributing to an adaptive LV hypertrophy, increase in angiogenesis and diminished cellular hypoxia. These mechanisms conceivably elicited the observed LV improvement of diastolic dysfunction caused by LPS challenge

Citrus hystrix leaf extract attenuated diabetic-cataract in STZ-rats.

Diabetic cataract causes severe vision loss. This study evaluated the effects of hesperidin-standardized Citrus hystrix leaf flavonoids-rich extract (CLE) on diabetic-cataract development. Streptozotocin-induced diabetic rats were orally given 150 and 300mg CLE/kg body-weight. These were compared with non-treated diabetic or healthy rats as controls, over 8weeks. The CLE gradually attenuated fasting blood glucose (FBG), biomarkers for inflammation (Tumor necrosis factor alpha TNF-α; prostaglandin E2 PGE2); vascular permeability, (Vascular endothelial growth factor VEGF); and oxidative stress, (malondialdehyde MDA). The diabetic cataract was significantly mitigated by the 150mg CLE/kg dose. Good correlations were found between cataract incidence with FBG (r2 =0.90), serum PGE2 (r2 =0.91), MDA (r2 =0.99), VEGF (r2 =0.71), but not with TNF-α levels (r2 =0.49) suggesting the serum FBG, PGE2, MDA, and possibly the VEGF levels may help to predict the cataract risks. The CLE mitigated cataract probably by attenuating hyperglycaemia, inflammation, lens fluid influx, vascular leakage, lens osmotic-imbalance, and fibers over-hydration. PRACTICAL APPLICATIONS: The study shows the flavonoids-rich Citrus hystrix leaf consumption, effectively attenuated diabetes (fasting blood glucose) and mitigated diabetic cataract. It help reduce diabetes-related hyperglycaemia, oxidative stress, inflammation, and vascular leakage. The evidences were the CLE consumptions reduced the serum biomarkers tumor necrosis factor-alpha TNF-α; prostaglandin E2 PGE2, vascular endothelial growth factor (VEGF), and malondialdehyde (MDA). The C. hystrix leaf contains hesperidin, apiin, diosmin, saponarin, apigetrin, rutin and xanthotoxol, and other flavonoid glucosides. The study also showed good correlations between cataract incidence with fasting blood glucose FBG (r2 =0.90), serum PGE2 (r2 =0.91), and MDA (r2 =0.99), and less closely with VEGF (r2 =0.71) suggesting these serum biomarkers may help predict cataract risks. The CLE indicated cataract mitigation properties probably by attenuating FBG, inflammation, lens fluid influx, lens osmotic-imbalance, and fibers over-hydration.

Osteopontin And Angiogenic Factors As New Biomarkers Of Prostate Cancer.

The novel biomarkers that would identify patients at risk for relapse and metastatic spread are needed. The aim of this study was the evaluation of serum levels of osteopontin (OPN) and tumor endogenous angiogenic factors such as vascular-endothelial growth factor (VEGF), vascular-endothelial growth factor receptor 2 (VEGF R2), endostatin, angiostatin and thrombospondin 1, in prostate cancer (PC) patients. Blood concentrations of the analyzed parameters were determined in 40 prostate cancer patients eligible for radiotherapy as well as in the control group consisted of 25 volunteers. Commercial ELISA kits were used for the analysis. Significantly higher levels of OPN (101.49 ng/mL vs 59.88 ng/mL; P<.001), endostatin (252.60 ng/mL vs. 223.55 ng/mL; P=.043), angiostatin (47 ng/mL vs. 13 ng/mL; P=.047), VEGF (262.1 pg/mL vs. 138.0 pg/mL; P=.056) and VEGF R2 (11188.81 pg/mL vs. 9377.50 pg/mL; P=.047) were detected in PC patients compared with the control group. In PC patients we showed a positive correlation between OPN level and TNM clinical stage(R=0.36; P=.02) and negative correlation between OPN level and hemoglobin concentration (R=-0.33; P=.04). The study showed higher levels of the angiogenic factors in PC patients compared with the control group and identified OPN as an indicator of the PC clinical stage as well as a decreased hemoglobin level.

Apatinib reverses alectinib resistance by targeting vascular endothelial growth factor receptor 2 and attenuating the oncogenic signaling pathway in echinoderm microtubule-associated protein-like 4-anaplastic lymphoma kinase fusion gene-positive lung cancer cell lines.

Overexpression of insulin growth factor 1 receptor (IGF-1R) and its ligand, insulin growth factor 1 (IGF-1), is related to treatment resistance and worse prognosis in many types of tumors. We reported recently that IGF-1R activation by IGF induces resistance to alectinib and stimulates the production of vascular endothelial growth factor, which indicates that IGF induces alectinib resistance and angiogenesis. This study aimed to determine the effect of bigeminal inhibition of anaplastic lymphoma kinase (ALK) and angiogenesis on human insulin growth factor 1 receptor (hIGF-1)-triggered drug resistance in echinoderm microtubule-associated protein-like 4-anaplastic lymphoma kinase (EML4-ALK)-positive lung cancer. Human lung adenocarcinoma H3122 and H2228 cells were exposed to a combination of insulin growth factor 1 receptor (IGF-1), alectinib, or apatinib. The effects of the combination therapy were examined using cell the Cell Counting Kit-8 assay, the colony-forming assay, the scratch test, and flow cytometry analysis, and the molecular mechanism was assessed by western blot. At nontoxic concentrations, apatinib restored alectinib sensitivity by increasing drug-induced apoptosis and inhibiting viability, migration, and invasion in IGF-triggered drug resistant cells. Moreover, we found that apatinib restored sensitivity to alectinib mainly through suppression of the ALK downstream signaling pathway and antiangiogenesis signaling. Taken together, our results indicate that simultaneous inhibition of ALK and vascular endothelial growth factor R2 by the combination of alectinib with apatinib may be useful for controlling progression of EML4-ALK fusion gene lung cancer by reversing ALK-TKI resistance and inhibiting angiogenesis.

Targeting expression of adenosine receptors during hypoxia induced angiogenesis – A study using zebrafish model

Hypoxia is known to be a major player during pathological angiogenesis and adenosine as a negative feedback signaling to maintain oxygen delivery in pathological ischemic condition. We mimicked hypoxic condition and studied angiogenesis by inducing adenosine receptors using forskolin, a plant compound and NECA analogue of adenosine using zebrafish model. Vascular endothelial growth factor (VEGF) is known to play a key role during pathological angiogenesis and regulated by the factors HIF1a under hypoxic condition and recently Notch is proposed to play a negative feedback loop mechanism along with VEGF signaling but the role of adenosine receptor during the process is not known. We evaluated the mRNA expression of adenosine receptors (A1, A2a.1, A2a.2, A2b), HIF1a, VEGF A, VEGF R2, NRP1a, NOTCH 1a and DLL4 and the phenotypic variations of zebrafish embryos when treated with DAPT, γ-secretase inhibitor of Notch in addition to treating the embryos with SU5416, a VEGF receptor inhibitor. Upregulation of adenosine receptors (A1, A2a.1, A2a.2, A2b), HIF1a, VEGF A, VEGF R2, NRP1a, NOTCH1a and DLL4 was observed embryos were when treated with forskolin and NECA could possibly mimic hypoxic condition. Hatching and heart rate also increased with NECA and forskolin. SU5416 showed decreases in blood vessel formation and decreased adenosine receptors, VEGF, VEGFR2, HIF1a and NRP1a expression and DAPT, exhibited decreases in blood vessels and decreased NRP1a, NOTCH1a, DLL4 expression. These embryos developed with poor vasculature, tail bending, abnormal phenotypes and developmental delay. Forskolin treated with inhibitors showed increased blood vessel formation, normal phenotype, development and adenosine receptors (A1, A2a.1, A2a.2, A2b), HIF1a, VEGF A, VEGF R2, NRP1a, NOTCH 1a and DLL4 gene expression suggesting that adenosine mediated Notch and VEGF could play an important role during development and angiogenesis. Targeting VEGF and Notch signaling with adenosine receptors inhibitors which might have a therapeutic significance during hypoxia and abnormal angiogenesis.

VEGF regulates hippocampal neurogenesis and reverses cognitive deficits in immature rats after status epilepticus through the VEGF R2 signaling pathway.

Epilepsy is the most common chronic disease in children, who exhibit a higher risk for status epilepticus (SE) than adults. Hippocampal neurogenesis is altered by epilepsy, particularly in the immature brain, which may influence cognitive development. Vascular endothelial growth factor (VEGF) represents an attractive target to modulate brain function at the neurovascular interface and is a double-edged sword in seizures. We used the lithium-pilocarpine-induced epilepsy model in immature Sprague-Dawley rats to study the effects of VEGF on hippocampal neurogenesis in the acute phase and on long-term cognitive behaviors in immature rats following status epilepticus (SE). VEGF correlates with cell proliferation in the immature brain after SE. By preprocessing VEGF in the lateral ventricles prior to the induction of the SE model, we found that VEGF increased the proliferation of neural stem cells (NSCs) and promoted the migration of newly generated cells via the VEGF receptor 2 (VEGFR2) signaling pathway. VEGF also inhibited cell loss and reversed the cognitive deficits that accompany SE. Based on our results, VEGF positively contributes to the initial stages of neurogenesis and alleviates cognitive deficits following seizures; moreover, the VEGF/VEGFR2 signaling pathway may provide a novel treatment strategy for epilepsy.

Role of NF-κB activation and VEGF gene polymorphisms in VEGF up regulation in non-proliferative and proliferative diabetic retinopathy.

The present study was aimed to investigate the relation between nuclear factor kappa beta (NFκB) activation and downstream up-regulation of vascular endothelial growth factor (VEGF) in diabetic retinopathy (DR). Moreover the study was intended to evaluate the role of VEGF gene single nucleotide polymorphisms (SNPs) in DR occurrence and to investigate the functional relevance of VEGF gene SNPs in terms of VEGF expression in DR. Serum level of VEGF, VEGF R1 (receptor 1), VEGF R 2 (receptor 2) and NFκB (p50/65) activity was measured by enzyme linked immune sorbent assay. Genotyping and allelic composition of different SNPs i.e., rs2010963, rs3025039, rs1570360 and rs 2071559 were investigated by Taqman SNP genotyping assay. VEGF, NFκB p50/p65, and VEGF R1 & R2 gene expressions were quantified by real time quantitative polymerase chain reaction. Increased NFκB p50/p65 activity and expressions were observed in non proliferative diabetic retinopathy (NPDR) and proliferative diabetic retinopathy (PDR) subjects compared to type 2 diabetes mellitus without retinopathy (DNR) group. Significantly elevated levels of serum VEGF and highest VEGF expression were found among PDR subjects compared to DNR or NPDR subjects. CC genotype and C allele of rs2010963 and TT genotype and T allele of rs3025039 were significantly over represented among PDR subjects compared to DNR group. Increased activation of NFκβ in NPDR and PDR subjects might involve increased up regulation of VEGF. VEGF SNPs i.e., rs2010963 C allele and rs3025039 T allele might be associated with PDR occurrence and in turn regulates VEGF expression among PDR subjects.

Proangiogenic compositions of microvesicles derived from human umbilical cord mesenchymal stem cells.

Introduction & ObjectiveMicrovesicles (MVs) derived from mesenchymal stem cells (MSCs) have been shown to promote angiogenesis. This study was aimed to shed a light on the mechanisms by analyzing the angiogenesis-promoting compositions of MSC-MVs. Also we try to figure out the impact of hypoxia on angiogenesis.MethodsMVs were isolated from the culture supernatants of MSCs under hypoxia/normoxia and serum-deprivation condition. The morphological features of MVs were revealed by an electron microscope and the origin of the MVs was identified by a bead-bound assay. An antibody array was used to analyze the expression of angiogenic cytokines from MVs and the parent MSCs as well. The major candidate factors were screened and the results were validated by immune blotting.ResultsMSC-MVs were around 80 nm in diameter. They expressed CD29, CD44, and CD73, but not CD31 and CD45. Antibody array showed that both MSCs and MVs expressed many angiogenesis-promoting biomolecules, including interleukin-6 (IL-6), basic fibroblast growth factors (bFGF), and recptor of urokinase-type plasminogen activator (UPAR). MSC-MVs contained angiogenin, vascular endothelial growth factor (VEGF), monocyte chemotactic protein-1 (MCP-1) and the receptor-2 for vascular endothelial growth factor at higher levels than the parent MSCs. Under hypoxic condition most cytokines were expressed in greater quantity than normoxic in MSCs while in MVs there was no significant difference between hypoxic and normoxic conditions except UPAR, Angiogenin, VEGF, IGF, Tie-2/TEK, and IL-6 which were higher in MVs under hypoxic conditions than those in normoxic condition.ConclusionUpon serum-deprivation condition, MSCs could secrete MVs that contain a variety of factors contributing to their angiogenesis-promoting function. And among them, Angiogenin, VEGF, MCP-1, VEGF R2 might be of greater importance than the other cytokines. Also UPAR, Angiogenin, VEGF, IGF, Tie-2/TEK, IL-6 might be responsible for hypoxia-augmented proangiogenic effects of MVs.

Abstract 16: Immunomodulatory effects of VEGF on human lymph node antigen-presenting and lymphoid cells

Abstract Background: Vascular endothelial growth factor (VEGF) is a potent pro-angiogenic signal associated with the growth and metastasis of tumors. In addition, VEGF has also been implicated in the suppression of immune cell function. The purpose of this study was to characterize VEGF secretion, receptor expression and effect of soluble VEGF neutralization in human lymph node cells both at rest and during activation. Methods: Human lymph nodes derived from patients with melanoma (IRB CASE 3610) containing T cells, B cells, monocyte/macrophages and dendritic cells were cultured with anti-CD3/CD28 beads and IL-2 in order to induce an inflammatory response in vitro. Culture supernatant was assayed for the presence of soluble VEGF on days 4, 7, 11 and 14 of culture. Multiparameter FACS analysis was performed to assess the cell surface expression of VEGF receptor subtypes 1 and 2 over the 14-day culture period. In addition, intracellular expression of cytokines within effector T cells was assessed at the end of the culture. Finally, a parallel culture containing a VEGF neutralizing antibody was performed to assess the effect of VEGF neutralization on effector T cell phenotype. High-throughput T cell receptor CDR3 sequencing was used to determine the molecular sequence of the variable region of the T cell receptors (ImmunoSEQ, Seattle, WA). Results: VEGF was measureable in the culture supernatant on days 4 and beyond. The proportion of cells expressing intracellular VEGF peaked at day 4 and subsequently decreased. CD11b+(monocyte/macrophage), CD11c+ (dendritic cells) and CD19+ (B) cells had the highest proportion of cells which expressed intracellular VEGF. VEGF R1 and R2 were not constitutively expressed on resting immune cells, but receptor expression was upregulated upon activation of the T cells in vitro. Day 14 T cells demonstrated intracellular IFN-γ, IL-2 and TNF-α. The addition of VEGF-blocking antibody in the culture of MDLN cells effectively neutralized the amount of soluble VEGF detected in the culture supernatant at all days. The addition of the VEGF-blocking antibody also increased the proportion of T cells expressing intracellular IFN-γ as compared T cells in the non-VEGF blocked culture and resulted in distinct molecularly defined T cell clones. Conclusion: These results confirm that soluble VEGF is produced by immune cells during activation, even in the absence of tumor. VEGF receptor expression on lymph node immune cell subsets is constitutive, but low, and then upregulated during in vitro activation of T cells, presumably due to secondary cytokine production. Neutralization of soluble VEGF resulted in qualitative differences in effector cytokine expression and T cell receptor sequences. Whether the effect on these T cells is direct via VEGF interaction with the cognate receptor or from an indirect effect on antigen-presenting cells is currently under investigation. Citation Format: Madeleine P. Strohl, Hallie Graor, Mei Zhang, Anthony Visioni, John Ammori, Isabelle Rivers-McCue, Julian Kim. Immunomodulatory effects of VEGF on human lymph node antigen-presenting and lymphoid cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 16. doi:10.1158/1538-7445.AM2014-16

A novel decoy receptor fusion protein for FGF-2 potently inhibits tumour growth.

Background:Antiangiogenic therapies have been proven effective in cancer treatment. Fibroblast growth factor-2 (FGF-2) has been functionally implicated in tumour angiogenesis and is an important target of antiangiogenic therapies. The aim of this work was to develop a novel FGF-2 inhibitor for cancer therapy.Methods:Eleven fusion proteins were developed by fusing various truncated extracellular regions of FGFR1 with the Fc region of IgG1. The optimal decoy receptor fusion protein with the highest binding affinity for FGF-2 was identified by an FGF-2-binding assay and its potential antitumour effects were investigated.Results:We obtained a soluble decoy receptor fusion protein with the highest binding activity for FGF-2, named FGF-Trap. Fibroblast growth factor-Trap significantly abolished FGF-2-stimulated activation of FGF signalling as demonstrated by its suppression of FGF-2-mediated phosphorylation of Erk1/2 and Akt, upregulation of cyclins D1 and E and the increase in mRNA levels of vascular endothelial growth factor R1 and R2 (VEGFR1 and VEGFR2). Furthermore, FGF-Trap effectively suppressed FGF-2-induced proliferation and migration of human umbilical vein endothelial cells (HUVECs) in vitro. Most importantly, FGF-Trap potently inhibited tumour growth and angiogenesis in Caki-1 and A549 xenograft models in vivo.Conclusions:Fibroblast growth factor-Trap potently inhibits tumour growth by blocking FGF-2 signalling pathways and could be an effective therapeutic agent for cancer patients.

Elevated blood levels of inflammation-related proteins are associated with an attention problem at age 24 mo in extremely preterm infants

Extremely preterm birth is associated with subsequent behavioral problems. We hypothesized that perinatal systemic inflammation, a risk factor for cerebral white matter injury and cognitive impairment, is associated with behavior problems observed at 2 y. In a cohort of 600 children born before 28 wk gestation, we measured 25 inflammation-related proteins in blood collected on postnatal days 1, 7, and 14, and identified behavior problems using parent responses to the Child Behavior Checklist for Ages 1.5-5 (CBCL/1.5-5) at 2 y of age. A persistent or recurrent protein elevation was defined as a concentration in the highest quartile (for gestational age and postnatal age) on at least 2 d ~1 wk apart. Behavior problems were defined by CBCL/1.5-5 subscale scores at or above the 93 rd percentile. A single-day elevation of intercellular adhesion molecule-3 was associated with an increased risk of an attention problem, as were persistent or recurrent elevations of myeloperoxidase, interleukin-6, tumor necrosis factor-RI, interleukin-8, intercellular adhesion molecule-3, vascular endothelial growth factor-R1, and vascular endothelial growth factor-R2. These associations persisted among infants without white matter injury and cognitive impairment. Among children born extremely prematurely, recurrent, or persistent elevations of inflammation-related proteins in blood during in the first two postnatal weeks are associated with an attention problem at age 2 y.

Correction of diabetic erectile dysfunction with adipose derived stem cells modified with the vascular endothelial growth factor gene in a rodent diabetic model.

The aim of this study was to determine whether adipose derived stem cells (ADSCs) expressing vascular endothelial growth factor (VEGF) gene can improve endothelial function, recover the impaired VEGF signaling pathway and enhance smooth muscle contents in a rat diabetic erectile dysfunction (DED) model. DED rats were induced via intraperitoneal injection of streptozotocin (40 mg/kg), and then screened by apomorphine (100 µg/kg). Five groups were used (n = 12/group)–Group 1 (G1): intracavernous injection of lentivirus-VEGF; G2: ADSCs injection; G3: VEGF-expressing ADSCs injection; G4: Phosphate buffered saline injection; G1–G4 were DED rats; G5: normal rats. The mean arterial pressure (MAP) and intracavernosal pressure (ICP) were measured at days 7 and 28 after the injections. The components of the VEGF system, endothelial, smooth muscle, pericytes markers in cavernoursal tissue were assessed. On day 28 after injection, the group with intracavernosum injection of ADSCs expressing VEGF displayed more efficiently and significantly raised ICP and ICP/MAP (p<0.01) than those with ADSCs or lentivirus-VEGF injection. Western blot and immunofluorescent analysis demonstrated that improved erectile function by ADSCs-VEGF was associated with increased expression of endothelial markers (VEGF, VEGF R1, VEGF R2, eNOS, CD31 and vWF), smooth muscle markers (a-actin and smoothelin), and pericyte markers (CD146 and NG2). ADSCs expressing VEGF produced a therapeutic effect and restored erectile function in diabetic rats by enhancing VEGF-stimulated endothelial function and increasing the contents of smooth muscle and pericytes.

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Introduction: Near-Infrared Spectroscopy (NIRS) may be utilized during a vascular occlusion test, where limb blood flow is temporarily occluded and released, to quantify a tissue bed’s ability to re-oxygenate by measuring STO2 recovery rate (upslope) during reperfusion. Hypothesis: The NIRS upslope measurement has been previously postulated to represent endothelial cell dysfunction. The hypothesis of this study is that NIRS upslope is associated with endothelial biomarkers, thus making it a feasible surrogate for assessing endothelial activation in patients with sepsis. Methods: A prospective, multicenter, observational study of a cohort of adult patients at four university emergency departments consisting of patients with septic shock (systolic blood pressure < 90 mmHg after fluid challenge), sepsis without shock and patients without infection. At enrollment we measured the NIRS derived upslope and collected plasma to assay biomarkers of endothelial activation, namely vascular endothelia growth factor (VEGF), soluble vascular endothelial growth factor R1 (sFLT-1), sE-Selectin, soluble intercellular adhesion molecule (sICAM-1), soluble vascular cell adhesion molecule (sVCAM-1) and tissue plasminogen activator inhibitor-1 (tPAI-1). Outcomes were organ dysfunction (measured as SOFA-score at 24 hours) and in-hospital mortality. Results: A total of 180 patients were included. Mean age was 63 (± 16) of whom 60% were males. Univariate analysis looking at the linear relationship for upslope prediction of endothelial biomarkers, we found a weak but statistical significant association between NIRS upslope and sFLT-1 and tPAI-1 (r=-0.08, p<0.0001 and r=-0.06, p=0.002). When adjusting for diabetes, age and SOFA-score at enrollment, only sFLT-1 persisted having a weak association (r=-0.04, p=0.01). In univariate subgroup analysis of the septic shock cohort (n=42) we found a statistical significant association between upslope and VEGF (r=0.11, p<0.05), which persisted after adjusting for diabetes, age and SOFA-score at enrollment (r=0.11, p<0.05). No association between sE-Selectin, sICAM-1 or sVCAM-1 were found in the analyses. Conclusions: NIRS upslope cannot be used as a surrogate marker for endothelial activation.

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Introduction: Our previous work suggests that markers of endothelial cell activation/dysfunction are associated with organ dysfunction and mortality in patients with sepsis. Hypothesis: The objective of this study is to investigate the association between endothelial markers with organ dysfunction (assessed by SOFA-score) and in-hospital mortality to validate our earlier findings in a multicenter study. We hypothesize that endothelial biomarkers, particularly sfLT-1 hold promise as novel prognostic markers in sepsis. Methods: This was a prospective, multicenter, observational study of a convenience sample consisting of three cohorts of adult patients at four university emergency departments: 1) a septic shock cohort (SBP<90mmHg after fluid challenge), 2) a sepsis without shock cohort, and 3) ED patients without infection. We collected plasma at enrollment, and then assayed biomarkers of endothelial activation, namely, soluble vascular endothelial growth factor R1 (sFLT-1), sE-Selectin, soluble intercellular adhesion molecule (sICAM-1), soluble vascular cell adhesion molecule (sVCAM-1), plasminogen activator inhibitor-1 (tPAI-1) and the inflammatory cytokine IL-6. Outcomes were organ dysfunction (measured as Sequential Organ Failure Assessment (SOFA) score at 24 hours) and in-hospital mortality. Results: We enrolled a total of 203 patients. The overall mean age was 63 (± 16) years, of whom 60% were males. There was a relationship between nearly all target biomarkers and organ dysfunction at 24 hours (sVEGF-R1 r=0.59, sVCAM-1 r=0.42, sE-Selectin r=0.37, PAI-1 r=0.32 p <0.0001 for each; and sICAM-1 r=0.16 and P <0.028). Among the endothelial biomarkers, sVEGF-R1 had the strongest association with SOFA score at 24 hours. sFLT-1 and TPAI-1 had the highest area under the receiver operator characteristic curves for mortality of 0.89 (95% CI 0.81-0.98) and 0.90 (0.82-0.97) respectively. For comparison IL-6 had an AUC of 0.82 (0.72-0.91). Conclusions: This multicenter validation study confirms that markers of endothelial cell activation are associated with organ dysfunction and mortality in sepsis.

Vascular endothelial growth factor enhances macrophage clearance of apoptotic cells

Efficient clearance of apoptotic cells from the lung by alveolar macrophages is important for the maintenance of tissue structure and function. Lung tissue from humans with emphysema contains increased numbers of apoptotic cells and decreased levels of vascular endothelial growth factor (VEGF). Mice treated with VEGF receptor inhibitors have increased numbers of apoptotic cells and develop emphysema. We hypothesized that VEGF regulates apoptotic cell clearance by alveolar macrophages (AM) via its interaction with VEGF receptor 1 (VEGF R1). Our data show that the uptake of apoptotic cells by murine AMs and human monocyte-derived macrophages is inhibited by depletion of VEGF and that VEGF activates Rac1. Antibody blockade or pharmacological inhibition of VEGF R1 activity also decreased apoptotic cell uptake ex vivo. Conversely, overexpression of VEGF significantly enhanced apoptotic cell uptake by AMs in vivo. These results indicate that VEGF serves a positive regulatory role via its interaction with VEGF R1 to activate Rac1 and enhance AM apoptotic cell clearance.

Effect of psoriasis therapy on VEGF and its soluble receptors serum concentrations

Vascular endothelial growth factor (VEGF) is recognized a pivotal pro-angiogenic factor responsible for new blood vessels formation in psoriatic lesion. The aim of the study was to analyse serum concentrations of VEGF and its soluble receptors (sVEGF) R1 and R2 in psoriatic patients before and after treatment. Serum samples were collected before and after 14 days of standard topical therapy, from 44 patients with exacerbated chronic plaque-type psoriasis and VEGF, sVEGF R1 and sVEGF R2 concentrations were measured using an enzyme immunoassay. Data were analysed with respect to baseline values of the psoriasis area severity index (PASI). Baseline mean serum levels of VEGF and sVEGF R1, but not sVEGF R2 were significantly higher in patients than in healthy controls. VEGF demonstrated significant correlation with PASI score. Treatment resulted in significant reduction of VEGF serum concentration, particularly in patients with severe course of the disease (PASI >20) and increase in sVEGF R1 concentration in patients with mild disease activity (PASI <10). Moreover, serum sVEGF R1 level after treatment termination was significantly higher in patients with mild than severe course of psoriasis. We confirmed the association between psoriasis activity and serum VEGF concentrations, which can be recognized as an indicator of the disease severity. However, the increase of serum sVEGF R1 concentrations can predict amelioration of clinical signs.

Effect of psoriasis activity on VEGF and its soluble receptors concentrations in serum and plaque scales

Vascular endothelial growth factor (VEGF) demonstrating pro-angiogenic activity promote new blood vessel formation in psoriatic lesions. The aim of this study was to evaluate the serum concentrations of VEGF, its soluble receptors (sVEGF R1 and R2) and VEGF content in scales of patients with psoriasis. To analyze possible association with activity of the disease, serum and scales from plaques were collected from 59 patients with exacerbated chronic plaque-type psoriasis. Mean concentrations of VEGF and sVEGF R1 in sera of patients were respectively two and four times higher than in healthy controls. Serum VEGF and sVEGF R1, but not sVEGF R2 demonstrated significant correlation with psoriasis area and severity index (PASI). There was also significant correlation between VEGF levels in serum and scales. Serum sVEGF R1 concentration was significantly elevated even in patients with low psoriasis activity (PASI < 10), whereas increase of serum VEGF became significant in patients with medium activity (PASI: 10–20). Levels of serum VEGF and sVEGF R1 were the highest in patients with PASI > 20. We confirmed association of both serum and scales VEGF concentrations with degree of psoriasis activity and demonstrated predominant increase of sVEGF R1 vs. VEGF in serum of patients with low psoriasis activity.