The effect of preexisting medical comorbidities on the preeclamptic phenotype: a retrospective cohort study.

Objective:To compare the effect of comorbidities on the phenotype and outcomes of preeclampsia.Methods: A matched retrospective cohort study of women delivering at a tertiary maternity center following a diagnosis of preeclampsia. We collected data on signs and symptoms, biochemical markers, and maternal and perinatal outcomes.Results:We studied 474 women; 158 women with and 316 without comorbidities. Compared to women without comorbidities, women with comorbidities delivered earlier. They suffered fewer maternal but more neonatal complications.Conclusion: Women with comorbidities receive earlier intervention than women without comorbidities, which may lead to fewer maternal complications but worse neonatal outcomes.

High-intensity interval training followed by postexercise cold-water immersion does not alter angiogenic circulating cells, but increases circulating endothelial cells.

High-intensity interval training (HIIT) induces vascular adaptations that might be attenuated by postexercise cold-water immersion (CWI). Circulating angiogenic cells (CAC) participate in the vascular adaptations and circulating endothelial cells (CEC) indicate endothelial damage. CAC and CEC are involved in vascular adaptation. Therefore, the aim of the study was to investigate postexercise CWI during HIIT on CAC and CEC and on muscle angiogenesis-related molecules. Seventeen male subjects performed 13 HIIT sessions followed by 15 min of passive recovery (n = 9) or CWI at 10 °C (n = 8). HIIT comprised cycling (8-12 bouts, 90%-110% peak power). The first and the thirteenth sessions were similar (8 bouts at 90% of peak power). Venous blood was drawn before exercise (baseline) and after the recovery strategy (postrecovery) in the first (pretraining) and in the thirteenth (post-training) sessions. For CAC and CEC identification lymphocyte surface markers (CD133, CD34, and VEGFR2) were used. Vastus lateralis muscle biopsies were performed pre- and post-training for protein (p-eNOSser1177) and gene (VEGF and HIF-1) expression analysis related to angiogenesis. CAC was not affected by HIIT or postexercise CWI. Postexercise CWI increased acute and baseline CEC number. Angiogenic protein and genes were not differently modulated by post-CWI. HIIT followed by either recovery strategy did not alter CAC number. Postexercise CWI increased a marker of endothelial damage both acutely and chronically, suggesting that this postexercise recovery strategy might cause endothelial damage. Novelty HIIT followed by CWI did not alter CAC. HIIT followed by CWI increased CEC. Postexercise CWI might cause endothelial damage.

Utilidad de los marcadores bioquímicos de preeclampsia / Usefulness of the biochemical markers of pre-eclampsia

La preeclampsia (PE) constituye una de las principales causas de mortalidad materna y perinatal en el mundo. En los países desarrollados, los estudios apuntan a un importante aumento de la incidencia de PE en la última década, en parte, por el aumento de la prevalencia, en la población general, de enfermedades que afectan a la función vascular, como la diabetes, la hipertensión crónica o la enfermedad renal. En el presente documento se lleva cabo una revisión actualizada de la PE. Se describen los criterios diagnósticos y la fisiopatología de la enfermedad. El objetivo principal del documento es revisar los nuevos marcadores bioquímicos que pueden ser de utilidad en la práctica clínica para la predicción y el diagnóstico de la PE, así como los distintos métodos mediante los cuales se puede llevar a cabo su determinación

Pre-eclampsia (PE) is one of the leading causes of maternal and perinatal mortality in the world. In developed countries, studies point to a significant increase in the incidence of PE in the last decade, partly due to the increase in the prevalence in the general population of diseases that affect vascular function, such as diabetes. chronic hypertension, or kidney disease. An updated review of PE is presented in this article. The diagnostic criteria and the pathophysiology of the disease are described. The main objective of the document is to review the new biochemical markers that may be useful in clinical practice for the prediction and diagnosis of PE, as well as the different methods by which yey can be determined

New biomarkers in endometriosis.

Endometriosis is a benign gynecological disorder which presents significant challenges in terms of diagnosis and management. Despite decades of research, there are no sufficiently sensitive and specific signs and symptoms nor blood tests for the clinical confirmation of endometriosis, which hampers prompt diagnosis and treatment. The huge majority of potential biomarkers has been discarded in research stage and very few have been translated to clinical practice. Serum CA-125 is the most studied and used one, but studies have shown its poor diagnostic performance. Several factors involved in the chronic inflammatory process of endometriosis, such as hormones, cytokines, chemokines, angiogenic factors, oxidative stress markers and others, have been implicated in the disease's pathogenesis and have been extensively studied, but not a single one has successfully been able to accurately identify the disease. MicroRNAs have emerged more recently but their utility to detect endometriosis remains uncertain. The search for a biomarker or a set of biomarkers is still open and may benefit from novel molecular biology and bioinformatics approaches to mine and uncover molecular signatures specifically associated with the disease.

The role of angiogenic factors in the management of preeclampsia.

Preeclampsia is a pregnancy disorder causing substantial maternal and fetal morbidity and mortality. In the UK, its diagnosis currently depends upon new onset hypertension and proteinuria. There is a clinical need for enhanced screening to prevent unnecessary resource use and improve outcomes. Here, the current practice in preeclampsia diagnosis will be summarized, with assessment of the evidence that angiogenic factors could improve its management. Although the combination of new onset hypertension and proteinuria define and hence diagnose the disorder, separately they are poorly predictive. Preeclampsia is ultimately a placental disease caused by syncytiotrophoblast dysfunction. The angiogenic factors placental growth factor, soluble fms-like tyrosine kinase 1 and soluble endoglin, all originating at least in part from the syncytiotrophoblast, are biomarkers with predictive potential for preeclampsia and related adverse outcomes. Recent work with the soluble fms-like tyrosine kinase 1/placental growth factor ratio has identified key measurement cutoffs, with one having a high negative predictive value for preeclampsia. The soluble fms-like tyrosine kinase 1/placental growth factor ratio seems particularly promising as a screening measure, able to predict accurately the short-term absence of preeclampsia and suggest the likelihood of adverse events within 4 weeks. The ratio could be used to allocate specific management plans to patients according to risk. An understanding of angiogenic factors may also lead to new therapeutic options for a condition currently only curable by delivery, although it must be remembered that the factors are markers of underlying syncytiotrophoblast stress, which would not be resolved by targeting them.

A pilot study to profile salivary angiogenic factors to detect head and neck cancers.

BACKGROUND: Early diagnosis of head and neck squamous cell carcinoma (HNSCCs) is an appealing way to increase survival rates in these patients as well as to improve quality of life post-surgery. Angiogenesis is a hallmark of tumor initiation and progression. We have investigated a panel of angiogenic factors in saliva samples collected from HNSCC patients and controls using the Bio-Plex ProTM assays. METHODS: We have identified a panel of five angiogenic proteins (sEGFR, HGF, sHER2, sIL-6Ra and PECAM-1) to be elevated in the saliva samples collected from HNSCC patients (n = 58) compared to a control cohort (n = 8 smokers and n = 30 non-smokers). RESULTS: High positive correlations were observed between the following sets of salivary proteins; sEGFR:sHER2, sEGFR:HGF, sEGFR:sIL-6Rα, sHER2:HGF and sHER2:sIL6Ra. A moderate positive correlation was seen between FGF-basic and sEGFR. CONCLUSION: We have shown that angiogenic factor levels in saliva can be used as a potential diagnostic biomarker panel in HNSCC.

Micro-RNA profile and proteins in peritoneal fluid from women with endometriosis: their relationship with sterility.

OBJECTIVE: To define the microRNA (miRNA) profile and its relationship with cytokines content in peritoneal fluid (PF) from endometriosis patients. DESIGN: Case-control study. SETTING: University hospital, research institute. PATIENT(S): One hundred twenty-six women with endometriosis (EPF) and 45 control women (CPF). MAIN OUTCOMES MEASURE(S): MiRNA arrays were prepared from six EPF and six CPF. Quantitative reverse transcription-polymerase chain reaction validation of nine selected miRNAs (miR-29c-3p, -106b-3p, -130a-3p, -150-5p, -185-5p, -195-5p, -451a, -486-5p, and -1343-5p) was performed. Vascular endothelial growth factor-A (VEGF-A), thrombospondin-1 (TSP-1), urokinase plasminogen activator (uPA), plasminogen activator inhibitor-1 (PAI-1), matrix metalloproteinase-3 (MMP3), tissue inhibitor of metalloproteinases type 1 (TIMP-1), interleukin (IL)-6, IL-8, IL-17A, macrophage inflammatory protein 1ß (MIP1beta), platelet-derived growth factor α-polypeptide A, and regulated on activation, normal T cell expressed and secreted (RANTES) were quantified by ELISA and MILLIPLEX. RESULT(S): MiRNA arrays showed 126 miRNAs differentially expressed (fold change ±1.2) (78 down-regulated, 48 up-regulated) in EPF. Validation showed higher levels of miR-106b-3p, -451a, -486-5p, IL-6, IL-8, uPA, and TIMP-1 in EPF. In menstrual phase, EPF presented up-regulation of miR-106b-3p, -130a-3p, -150-5p, -185-5p, -451a, -486-5p, VEGF-A, IL-8, MIF 1ß, uPA, and PAI-1 compared with other phases; however, CPF did not. MiRNA-486-5p was up-regulated in sterile EPF compared with sterile controls, and VEGF-A, IL-8, and TIMP-1 were increased in sterile and fertile EPF compared with fertile CPF. CONCLUSION(S): MiRNAs seem to be involved in the peritoneal alterations in endometriosis, suggesting new mechanisms by which ectopic lesions could implant in endometriosis patients; and to serve as biomarkers for fertility outcome prediction.

Análise Dopplerfluxométrica e angiogênica de tumores mamários caninos / Dopplerfluxometric and angiogenic analysis of canine mammary tumors

Foi avaliado o comportamento de índices Doppler e a expressão de genes relacionados à neovascularização tumoral, visando caracterizar a vascularização das massas neoplásicas. Foram utilizadas 27 cadelas, com diagnóstico histopatológico de neoplasia mamária, sendo submetidas à avaliação Dopplerfluxométrica tumoral e à coleta de fragmentos neoplásicos para análise de expressão gênica de VEGF, FLT-1, FLK-1 e ATR1. Foram encontrados 22 tumores de origem epitelial (carcinomas) e cinco de origem mesenquimal (sarcomas). Observou-se correlação positiva entre o FLT-1 e as variáveis PS, PI e RI. O FLK-1 apresentou correlação igualmente positiva com os parâmetros PS e PI e uma tendência para RI, enquanto o VEGF retratou correlação positiva apenas com IP. O VEGF também mostrou correlação positiva com seus receptores, porém não apresentou correlação com o ATR1. O FLT-1 e o FLK-1 apresentaram ainda correlação positiva entre si e com o ATR1. Houve maior expressão média do VEGF nos tumores epiteliais do que nos mesenquimais. As variáveis PS, PI e RI, associadas com a expressão do VEGF e seus receptores, mostraram-se relevantes para caracterizar a neovascularização de tumores malignos, e a expressão diferenciada do VEGF entre os tipos tumorais pode ser um indicador auxiliar na caracterização de neoplasias mamárias malignas em cadelas.(AU)

The behavior of the tumor Doppler indexes and gene expression related to neovascularization was evaluated aiming to improve the characterization of neoplastic masses vascularization. Twenty-seven bitches with histopathological diagnosis of mammary neoplasia were submitted to tumor Dopplerfluxometric evaluation and collection of neoplastic fragments to analyze the gene expression of VEGF, FLT-1, FLK-1 and ATR1. Were found 22 epithelial (carcinomas) and five mesenchymal (sarcomas) tumors. Positive correlation was observed between FLT-1 and PS, PI and RI. FLK-1 presented a similar positive correlation with the PS and PI parameters, and a tendency for RI (r= 0.45, P= 0.07), whereas VEGF showed a positive correlation just with PI. VEGF also showed a positive correlation with its receptors, but did not present a correlation with ATR1. FLT-1 and FLK-1 also showed positive correlation with each other, and with ATR1. There was higher mean expression of VEGF in epithelial tumors than in mesenchymal ones. The PS, PI and RI associated with the expression of VEGF and its receptors have been shown to be relevant to characterize neovascularization of malignant tumors, and the differentiated expression of VEGF between the types of mammary tumors, may be an auxiliary indicator in the characterization of malignant breast cancers in bitches.(AU)

Heterogeneous histomorphology, yet homogeneous vascular smooth muscle cell dedifferentiation, characterize human aneurysm disease.

OBJECTIVE: Abdominal aortic aneurysm (AAA) is a frequent, potentially life-threatening, disease that can only be treated by surgical means such as open surgery or endovascular repair. No alternative treatment is currently available, and despite expanding knowledge about the pathomechanism, clinical trials on medical aneurysm abrogation have led to inconclusive results. The heterogeneity of human AAA based on histologic examination is thereby generally neglected. In this study we aimed to further elucidate the role of these differences in aneurysm disease. METHODS: Tissue samples from AAA and popliteal artery aneurysm patients were examined by histomorphologic analysis, immunohistochemistry, Western blot, and polymerase chain reaction. The results were correlated with clinical data such as aneurysm diameter and laboratory results. RESULTS: The morphology of human AAA vessel wall probes varies tremendously based on the grade of inflammation. This correlates with increasing intima/media thickness and upregulation of the vascular endothelial growth factor cascade but not with any clinical parameter or the aneurysm diameter. The phenotypic switch of vascular smooth muscle cells occurred regardless of the inflammatory state and expressional changes of the transcription factors Kruppel-like factor-4 and transforming growth factor-ß lead to differential protein localization in aneurysmal compared with control arteries. These changes were in similar manner also observed in samples from popliteal artery aneurysms, which, however, showed a more homogenous phenotype. CONCLUSIONS: Heterogeneity of AAA vessel walls based on inflammatory morphology does not correlate with AAA diameter yet harbors specific implications for basic research and possible aneurysm detection.

Angiogenic proteins, placental weight and perinatal outcomes among pregnant women in Tanzania.

INTRODUCTION: Placental vascular development, and ultimately placental weight, is essential to healthy fetal development. Here, we examined placental weight in a cohort of Tanzanian women in association with angiogenic proteins known to regulate placental vascular development and perinatal outcomes. METHODS: A total of n = 6579 women with recorded placental weight were included in this study. The relative risk of adverse perinatal outcomes (Apgar score, death, asphyxia, respiratory distress, seizures, pneumonia and sepsis) was compared between placental weight in the bottom and top 10th percentiles. We quantified angiogenic mediators (Ang-1, Ang-2, VEGF, PGF and sFlt-1) in plasma samples (n = 901) collected between 12 to 27 weeks of pregnancy using ELISA and assessed the relative risk of placental weight in the bottom and top 10th percentiles by protein levels in quartiles. RESULTS: Women with Ang-2 levels in the highest quartile had an increased relative risk of placental weight in the bottom 10th percentile (RR = 1.45 (1.10, 1.91), p = 0.01). Women with VEGF-A (RR = 0.73 (0.56, 0.96), p = 0.05) and PGF (RR = 0.58 (0.44, 0.72), p = 0.002) in the highest quartile had a reduced relative risk of placental weight in the bottom 10th percentile. Low placental weight (in bottom 10th percentile) was associated with an increased relative risk of Apgar score of <7 at 1 minute (RR = 2.31 (1.70, 3.13), p = 0.001), at 5 minutes (RR = 3.53 (2.34, 5.33), p = 0.001), neonatal death (RR = 5.02 (3.61, 7.00), p = 0.001), respiratory distress (RR = 4.80(1.71, 13.45), p = 0.001), and seizures (RR = 4.18 (1.16, 15.02), p = 0.03). DISCUSSION: The association between low placental weight and risk of adverse perinatal outcomes in this cohort suggests that placental weight could serve as a useful indicator, providing additional insight into high-risk pregnancies and identifying neonates that may require additional monitoring and follow-up.

Differences between perivascular adipose tissue surrounding the heart and the internal mammary artery: possible role for the leptin-inflammation-fibrosis-hypoxia axis.

AIM: The factors mediating the paracrine effects of perivascular adipose tissue (PVAT) in atherosclerosis are largely unknown. The adipokine leptin has been implicated in the increased cardiovascular risk in obesity and may locally promote neointima formation independently of circulating leptin levels. In patients with established coronary artery disease, we examined the expression of leptin as well as of its possible inducers in 'cardiac' PVAT surrounding the aortic root and coronary arteries (C-PVAT), and compared it to the PVAT surrounding the internal mammary artery (IMA-PVAT), a vessel resistant to atherosclerosis. METHODS AND RESULTS: Tissue specimens collected from male patients undergoing coronary artery bypass surgery were processed for real-time PCR, ELISA, in situ hybridization, and immunohistochemistry analysis. Leptin protein expression was elevated in C-PVAT compared to IMA-PVAT, independent of serum leptin levels. Compared to IMA-PVAT, C-PVAT exhibited more pronounced angiogenesis and inflammation, as indicated by significantly higher numbers of PECAM1-positive vessels and CD68-positive macrophages, and was characterized by a greater extent of fibrosis and hypoxia. Increased expression of hypoxia-inducible factor-1α and Fos-like antigen (FOSL)2, factors known to enhance leptin gene transcription, was observed in C-PVAT. As a proof of concept, exposure of human adipocytes to chemical hypoxia resulted in significantly increased FOSL2 and leptin mRNA levels. CONCLUSIONS: A higher degree of local tissue hypoxia and up-regulation of leptin expression in the perivascular adipose tissue, along with increased vascularization, inflammation, and fibrosis, may contribute to the increased atherosclerotic plaque burden in the coronary arteries compared to the IMA.

Altered Expression of Angiotensinogen and Mediators of Angiogenesis in Ileal Crohn's Disease.

BACKGROUND AND AIMS: Angiotensin II (AII) is a powerful splanchnic vasoconstrictor with pro-inflammatory and pro-fibrotic properties. Angiotensin converting enzyme (ACE) inhibitors and AII Receptor Antagonists (ARBs) are therapeutic in animal models of colitis. The aim of this case-control study is to determine the expression of angiotensinogen and related genes in human ileal Crohn's disease. METHODS: Using quantitative real-time polymerase chain reaction (RT-PCR), we measured mRNA expression levels of angiotensinogen (AGT), hypoxia inducible factor (HIF)1α and melanoma cell adhesion molecule (MCAM; CD146) in 101 human samples (69 biopsy, 12 resection) from affected ileum (inflamed CD cases, n=36) and unaffected ileum (non-inflamed CD cases, n=45 and controls, n=20). Immunohistochemistry for angiotensinogen was also performed. The study was of case-control design in a tertiary care setting. RESULTS: Ileal expression of AGT was lower in CD cases compared to controls (p<0.0001), in inflamed CD samples (p=0.017) and current smokers (p=0.02). HIF1α expression was lower in non-inflamed CD biopsy samples than controls (p=0.006). The presence of disease-associated NOD2 variants was associated with increased expression of markers of angiogenesis (HIF1α p=0.009; MCAM p=0.007) in inflamed CD samples. Angiotensinogen immunohistochemical staining supported the quantitative RT-PCR expression findings. CONCLUSIONS: Angiotensinogen expression is down regulated in human ileal CD, particularly in the presence of inflammation and current cigarette smoking, implicating the mesenteric vasculature and mucosal hypoxia as co-factors in ileal CD pathogenesis. A novel reduction in HIF1α expression in non-inflamed ileal mucosa in CD patients was also demonstrated.

Increasing demonstration of angiogenic markers in skin neoplastic lesions.

BACKGROUND: Skin cancer represents the most common worldwide malignancy. Angiogenesis is an important factor in tumor growth and metastasis. Given these facts, the purpose of the current study was to compare the levels of angiogenic proteins in the context of the most common malignant and premalignant skin lesions. METHODS: Immunohistochemistry of CD31, HIF1A, VEGFR1 and VEGFR2 was performed in basal cell carcinoma (BCC), actinic keratosis (AK) and squamous cell carcinoma of the skin (SCCS). RESULTS: SCCS presented with increased levels of HIF1A, VEGFR1 and VEGFR2 in comparison to AK. In addition, SCCS also demonstrated increased levels of HIF1A to BCCLR or BCCHR. BCC presented with more vessels than AK. However, no correlation was observed among CD31, HIF1A, VEGFR1 and VEGFR2. CONCLUSIONS: SCCS presented with higher levels of HIF1A, VEGFR1 and VEGFR2, while BCC demonstrated an increased number of vessels in relation to AK. These data suggest that antiangiogenic therapy might be useful for skin cancer treatment.

Serine-arginine protein kinase 1 (SRPK1), a determinant of angiogenesis, is upregulated in prostate cancer and correlates with disease stage and invasion.

Vascular endothelial growth factor (VEGF) undergoes alternative splicing to produce both proangiogenic and antiangiogenic isoforms. Preferential splicing of proangiogenic VEGF is determined by serine-arginine protein kinase 1 (SRPK1), which is upregulated in a number of cancers. In the present study, we aimed to investigate SRPK1 expression in prostate cancer (PCa) and its association with cancer progression. SRPK1 expression was assessed using immunohistochemistry of PCa tissue extracted from radical prostatectomy specimens of 110 patients. SRPK1 expression was significantly higher in tumour compared with benign tissue (p<0.00001) and correlated with higher pT stage (p=0.004), extracapsular extension (p=0.003) and extracapsular perineural invasion (p=0.008). Interestingly, the expression did not correlate with Gleason grade (p=0.21), suggesting that SRPK1 facilitates the development of a tumour microenvironment that favours growth and invasion (possibly through stimulating angiogenesis) while having little bearing on the morphology or function of the tumour cells themselves.

[Prognostic value of clinical and morphological characteristics in radiation and combined treatment for tongue cancer].

It is now possible to identify several key factors that determine biological characteristics of squamous cell cancer of the head and neck: genes p53, p16, cyclin D1, P13-K/Akt connected with metastasis proteins (proteases, proteins mesenchymal cells, cell adhesion molecules chemokines), angiogenesis factors (VEGF, PDGF, FGF, TGF-alpha and TGF-beta), IL-8; epidermal growth factor receptors. An important role of tumor cells plays microenvironment. Of course the above mentioned is only a small part of the factors that determine the livelihoods and the activity of cancer cells. All of these factors are potential predictors of the effectiveness of radiation and chemoradiation treatment and actively studied in recent decades.

Histologic variability in solitary fibrous tumors reflects angiogenic and growth factor signaling pathway alterations.

This study aimed to evaluate expression of receptor tyrosine kinases, their ligands, and mutational status in solitary fibrous tumors, with correlation to histopathologic variants, tumor stage, and aggressive behavior. Immunohistochemical staining for PDGFα; PDGFß; PDGFR-α; PDGFR-ß; IGF1R; EGFR; VEGF; IGF2; c-Met; c-kit; c-erbB2; PTEN; and phosphorylated (p)AKT, pS6, and p4EBP1 was analyzed in 114 cases of solitary fibrous tumor using tissue microarray. Mutational analysis was performed using Sequenom MassARRAY-based platform. Multiple growth factors were overexpressed in most tumors, and increased numbers of overexpressed factors correlated with activation of the AKT pathway as measured by increased expression of p4EBP1(P = .0005). Compared to hypocellular tumors, localized hypercellular tumors were associated with high vascular endothelial growth factor (32% versus 8%; P = .008) and PDGFß (41% versus 13%; P = .008). Metastatic tumors more frequently overexpressed PDGFR-α compared to localized tumors (75% versus 31%; P < .001). None of the factors examined had prognostic significance in primary tumors. Single-nucleotide polymorphisms involving MET were identified in 4 patients; these do not appear to drive tumor behavior and were not reflected in c-Met expression levels. Simultaneous overexpression of multiple growth factors is common in solitary fibrous tumors; variability in expression may contribute to tumor phenotype and aggressive behavior.

Sensitive, quantitative, and high-throughput detection of angiogenic markers using shape-coded hydrogel microparticles.

Elevated serum concentrations of angiogenic markers including vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), and platelet-derived growth factor (PDGF) have been correlated with various clinical disorders including cancer, cardiovascular diseases, diabetes mellitus, and liver fibrosis. In addition, the correlation between the serum concentrations of these factors, clinical diagnosis, prognosis, and response to therapeutic agents is significant. Thereby suggesting high-throughput detection of serum levels of angiogenic markers has important implications in early detection of different clinical disorders as well as for subsequent therapy monitoring. Here, we demonstrate the feasibility of utilization of shape-coded hydrogel microparticle based suspension arrays for quantitative and reproducible measurement of VEGF, FGF, and PDGF in single and multiplexed assays. Bio-inert PEG hydrogel attenuated the background signal thereby improving the sensitivity of the detection method as well as eliminating the need for blocking the proteins. In the singleplexed assay, the detection limits of 1.7 pg ml(-1), 1.4 pg ml(-1), and 1.5 pg ml(-1) for VEGF, FGF, and PDGF respectively indicated that the sensitivity of the developed method exceeds that of the conventional technologies. We also demonstrated that in the multiplexed assays, recovery of the proteins was within 20% of the expected values. The practical applicability of the hydrogel microparticle based detection system was established by demonstrating the ability of the system to quantify the production of VEGF, FGF, and PDGF by breast cancer cells (MDA-MB-231).

Effects of platelet-rich plasma and cell coculture on angiogenesis in human dental pulp stem cells and endothelial progenitor cells.

INTRODUCTION: Platelet-rich plasma (PRP) has been described as platelet concentrate. Growth factors released by activated platelets can improve wound vasculogenesis and enhance wound healing. In this study, we used PRP instead of serum to culture human dental pulp stem cells (hDPSCs) and endothelial progenitor cells (EPCs) and investigated revascularization ability. The effect of hDPSC and EPC coculture on vasculogenesis was also studied. METHODS: PRP was prepared by secondary centrifugation. Real-time polymerase chain reaction and Western blotting were used to determine the expression of vasculogenesis-related factors vascular endothelial growth factor, platelet-derived growth factor, fetal liver kinase 1 (Flk-1), and stromal cell-derived factor 1 (SDF-1) in cultured hDPSCs and EPCs. The cells were divided into 4 groups: EPCs + 10% fetal bovine serum (FBS), EPCs + 10% PRP, EPCs + hDPSCs + 10% FBS, and EPCs + hDPSCs + 10% PRP. Then, the formation of vessel-like structures was tested by the tube formation assay. RESULTS: On day 3, the expression levels of all the markers in the coculture groups were much higher than in the single-culture groups and were also higher in the PRP groups compared with the FBS groups (P < .05), except for SDF-1. Expression levels were significantly higher in the experimental groups (EPCs + 10% PRP, EPCs + hDPSCs + 10% FBS, and EPCs + hDPSCs + 10% PRP) than in the control group (EPCs + 10% FBS) and in the PRP groups/coculture groups compared with the FBS groups/single-culture groups (P < .01). The tube formation assay showed the area of vessel-like structures formed by the PRP group to be larger than in the FBS group (P < .05). CONCLUSIONS: PRP and coculture can both promote vasculogenesis, and PRP can promote EPCs to form vessel-like structures.

Angiogenesis and remodelling in human thoracic aortic aneurysms.

AIMS: Human thoracic aneurysm of the ascending aorta (TAA) is a chronic disease characterized by dilatation of the aortic wall, which can progress to vessel dissection and rupture. TAA has several aetiologies, but all forms present common features, including tissue remodelling. Here, we determined and characterized the angiogenic process associated with TAA and its relation with wall remodelling. METHODS AND RESULTS: Immunostaining for blood vessels showed an increased density of microvessels originating from the adventitia in the external medial layer of TAA compared with healthy aortas. Proteomic array analysis of 55 angiogenic factors in medial and adventitial layers showed different expression profiles in both tissue compartments between aneurysmal and healthy aortas. Quantification by ELISA confirmed that all forms of TAA contained higher levels of several pro- and anti-angiogenic factors, including angiopoietin-1 and -2, fibroblast growth factor-acidic, and thrombospondin-1, than that of healthy aortas. However, all groups showed comparable levels of vascular endothelial growth factor-A. Quantitative RT-PCR demonstrated that angiopoietins were overexpressed in TAA media. Immunostaining and electron microscopy revealed that neovessels had defective endothelial junctions and poor mural cell coverage. This incomplete structure was associated with the accumulation of plasminogen and albumin in the media of TAA. CONCLUSION: We describe, for the first time, leaky neovessel formation in TAA media in association with an imbalance of angiogenic factor levels. Although the initiating mechanisms of neo-angiogenesis in TAA and the potential aetiology-related differences remain to be determined, our results suggest that neo-angiogenesis could participate in TAA wall remodelling and weakening through deposition of blood-borne zymogens.

Serous neoplasms of the pancreas share many, but not all aspects of their microvascular and angiogenic profile with low-grade clear cell renal cell carcinomas.

Similarly to clear cell renal cell carcinomas (CCRCC), serous neoplasms (SN) of the pancreas frequently show inactivation of VHL gene, clear cell histology and abundant microvasculature. Data on the microvascular and angiogenic profile of SN are scarce. Aiming to examine further the striking resemblance of clear cell epithelial neoplasia in pancreas and kidney, we compared the microvascular profile and expression of pro-angiogenic factors in SN and in CCRCC using immunohistochemical stains. SN and CCRCC shared a predominance of differentiated blood vessels, scarcity of lymphatic vessels, presence of CD105 and claudin-5 in tumoral vessels, expression of vascular endothelial growth factor (VEGF)-A, cyclooxygenase-2 (COX-2), carbonic anhydrase IX in tumoral cells, and lack of VEGF-C in tumoral cells. In contrast to CCRCC, SN showed lower pericyte coverage of vessels, lower blood vessel endothelial cell proliferaction fraction, more pronounced VEGF receptor (VEGFR)-2 and glucose transporter-1 expression, higher inducible (iNOS) but lower endothelial nitric oxide synthase (eNOS) expression, as well as presence of VEGFR-3 and D2-40 expression in epithelial cells. In conclusion, we found a significant similarity but not equality of microvascular biology of SN and CCRCC. We recognized VEGFR-2, VEGFR-3, COX-2, iNOS, eNOS and D2-40 as new markers of epithelial cells of SN of the pancreas.