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Navarro A.,Fundacion Hospital General Universitario | Marin S.,Consorcio Hospital General Universitario | Minana M.D.,Fundacion Hospital General Universitario
Stem Cell Research and Therapy | Year: 2014

Introduction. Adipose tissue has the unique property of expanding throughout adult life, and angiogenesis is required for its growth. However, endothelial progenitor cells contribute minimally to neovascularization. Because myeloid cells have proven to be angiogenic, and monocytes accumulate in expanding adipose tissue, they might contribute to vascularization. Methods. The stromal vascular fraction (SVF) cells from human adipose tissue were magnetically separated according to CD45 or CD14 expression. Adipose-derived mesenchymal stromal cells (MSCs) were obtained from SVF CD45- cells. CD14+ monocytes were isolated from peripheral blood (PB) mononuclear cells and then cultured with SVF-derived MSCs. Freshly isolated or cultured cells were characterized with flow cytometry; the conditioned media were analyzed for the angiogenic growth factors, angiopoietin-2 (Ang-2), vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), granulocyte colony-stimulating factor (G-CSF), and granulocyte macrophage colony-stimulating factor (GM-CSF) with Luminex Technology; their angiogenic capacity was determined in an in vivo gelatinous protein mixture (Matrigel) plug angiogenesis assay. Results: CD45+ hematopoietic cells within the SVF contain CD14+ cells that co-express the CD34 progenitor marker and the endothelial cell antigens VEGF receptor 2 (VEGFR2/KDR), VEGFR1/Flt1, and Tie2. Co-culture experiments showed that SVF-derived MSCs promoted the acquisition of KDR and Tie-2 in PB monocytes. MSCs secreted significant amounts of Ang-2 and HGF, but minimal amounts of bFGF, G-CSF, or GM-CSF, whereas the opposite was observed for SVF CD14 + cells.Additionally, SVF CD14+ cells secreted significantly higher levels of VEGF and bFGF than did MSCs. Culture supernatants of PB monocytes cultured with MSCs contained significantly higher concentrations of VEGF, HGF, G-CSF, and GM-CSF than did the supernatants from cultures without MSCs. Quantitative analysis of angiogenesis at 14 days after implantation demonstrated that neovascularization of the implants containing SVF CD14+ cells or PB monocytes previously co-cultured with MSCs was 3.5 or 2 times higher than that observed in the implants with SVF-derived MSCs. Moreover, immunofluorescence of Matrigel sections revealed that SVF CD14 + cells differentiated into endothelial cells and contributed to vascular endothelium. Conclusions: The results from this study suggest that adipose tissue-resident monocytes should contribute to tissue vascularization. Because SVF CD14+ cells were more efficient in inducing angiogenesis than SVF-derived MSCs, and differentiated into vascular endothelial cells, they may constitute a new cell source for cell-based therapeutic angiogenesis. © 2014 Navarro et al.; licensee BioMed Central Ltd.

Izquierdo M.,Hospital Clinic Universitari | Ruiz-Granell R.,Hospital Clinic Universitari | Ferrero A.,Hospital Clinic Universitari | Martinez A.,Hospital Clinic Universitari | And 5 more authors.
Europace | Year: 2012

AimsElectrical storm (ES) is a life-threatening condition that predicts bad prognosis. Treatment includes antiarrhythmic drugs (AAD) and catheter ablation (CA). The present study aims to retrospectively compare prognosis in terms of survival and ES recurrence in 52 consecutive patients experiencing a first ES episode.Methods and resultsPatients were admitted from 1995 to 2011 and treated for ES by conservative therapy (pharmacological, 29 patients) or by CA (23 patients), according to the physician's preference and time of occurrence, i.e. conservative treatments were more frequently administered during the first years of the study, as catheter ablation became more frequent as the years passed by. After a median follow-up of 28 months, no differences either in survival (32 vs. 29 P 0.8) or in ES recurrence (38 in ablated vs. 57 in non-ablated patients, P 0.29) were observed between groups. Low left ventricle ejection fraction (LVEF) was the only variable associated with ES recurrence in ablated patients. When including patients with LVEF > 25, ES recurrence was significantly lower in ablated patients (24 months estimated risk of ES recurrence was 21 vs. 62 in ablated and non-ablated patients, respectively); however, no benefit in survival was observed.ConclusionOur data suggest that in most patients, especially those with an LVEF > 25, catheter ablation following a first ES episode, decreases the risk of ES recurrence, without increasing survival. © 2012 Published on behalf of the European Society of Cardiology. All rights reserved. © The Author 2012. For permissions please email: journals.permissionsoup.com.

Roig J.V.,Consorcio Hospital General Universitario | Cantos M.,Consorcio Hospital General Universitario | Balciscueta Z.,Hospital Universitario Arnau Of Vilanova | Uribe N.,Hospital Universitario Arnau Of Vilanova | And 5 more authors.
Colorectal Disease | Year: 2011

Aim The study evaluated the rate of reversal of Hartmann's operation after the initial surgery and its morbidity. Method A multicentre retrospective study was carried out in seven hospitals in the Valencia area of patients who underwent Hartmann's operation from 2004 to 2008. The incidence of reversal was determined. Results Four hundred and fifty-two patients of mean age 67.5±15.4years were included, of whom 78.8% had an emergency operation. The most common diagnosis was cancer (58.6%), although diverticulitis predominated in the emergency setting. At a median follow up of 44months, 159 (35.2%) patients had undergone reversal, including 16.6% after elective surgery and 40.4% after an emergency Hartmann's procedure (P<0.001). The most frequent reason why reversal was not done was death (74 [25%] patients). Patients undergoing reversal were younger and had a low ASA risk. Trauma was associated with a higher rate of reversal, followed by diverticular disease. Surgery was performed at a median of 10months. An open approach with stapled anastomosis was used in most cases. The mortality was 3.5%. Complications occurred in 45.2%, with a 6.2% rate of anastomotic leakage. Complications were associated with age, diabetes mellitus, arteriosclerosis, obesity, smoking, chemotherapy and COPD. Conclusion Hartmann's reversal was performed in a small percentage of patients, mostly including those with benign disease. It had a significant morbidity. © 2011 The Authors. Colorectal Disease © 2011 The Association of Coloproctology of Great Britain and Ireland.

Milara J.,Consorcio Hospital General Universitario | Milara J.,Polytechnic University of Valencia | Milara J.,Research Foundation | Peiro T.,Research Foundation | And 7 more authors.
Thorax | Year: 2013

Background: Cigarette smoking contributes to lung remodelling in chronic obstructive pulmonary disease (COPD). As part of remodelling, peribronchiolar fibrosis is observed in the small airways of patients with COPD and contributes to airway obstruction. Epithelial to mesenchymal transition (EMT) appears to be involved in the formation of peribronchiolar fibrosis. This study examines the EMT process in human bronchial epithelial cells (HBECs) from non-smokers, smokers and patients with COPD as well as the in vitro effect of cigarette smoke extract (CSE) on EMT. Methods: HBECs from non-smokers (n=5), smokers (n=12) and patients with COPD (n=15) were collected to measure the mesenchymal markers α-smooth muscle actin, vimentin and collagen type I and the epithelial markers E-cadherin, ZO-1 and cytokeratin 5 and 18 by real time-PCR and protein array. In vitro differentiated bronchial epithelial cells were stimulated with CSE. Results: Mesenchymal markers were upregulated in HBECs of smokers and patients with COPD compared with non-smokers. In contrast, epithelial cell markers were downregulated. In vitro differentiated HBECs underwent EMT after 72 h of CSE exposure through the activation of intracellular reactive oxygen species, the release and autocrine action of transforming growth factor β1, the phosphorylation of ERK1/2 and Smad3 and by the downregulation of cyclic monophosphate. Conclusions: The EMT process is present in bronchial epithelial cells of the small bronchi of smokers and patients with COPD and is activated by cigarette smoke in vitro.

Escutia B.,Hospital Universitario La Paz | Ledesma E.,Instituto Valenciano Of Oncology | Serra-Guillen C.,Instituto Valenciano Of Oncology | Gimeno C.,Consorcio Hospital General Universitario | And 3 more authors.
Journal of the European Academy of Dermatology and Venereology | Year: 2011

Background: Human papilloma virus (HPV) is increasingly recognized as an important human carcinogen but its role in the aetiopathogenesis of basal cell carcinoma (BCC) in immunocompetent individuals is unclear. Objective: A prospective case-control study was designed to compare the prevalence of HPV in BCC and normal skin samples from immunocompetent subjects and to assess the influence of different clinical features on the risk of cutaneous HPV. Methods: A total of 142 samples from 70 BCC cases (superficial BCC 38 and nodular BCC 32) and 72 controls were analysed by a degenerated nested PCR technique. Clinical data were recorded and risk factors for HPV infection were assessed by multivariate analysis. Results: There were 31 HPV DNA-positive samples. HPV was detected more frequently in cases (25.7%) than in controls (18.1%) and in nodular (31.3%) than in superficial (21.1%) BCC lesions but differences were not statistically significant. Older age (OR 1.5; 95% CI: 1.02-1.09) and actinic keratosis (OR 2.62; 95% CI 1.15-5.96) were the only significant factors associated to the presence of HPV. Risk of HPV positivity was also higher in blond-haired subjects, fair/pale skin colour, history of sunburn, solar lentigines and seborrheic keratosis but the differences were not significant. Both in cases and controls, b -types were the most frequent. Conclusions: HPV does not seem to play a fundamental role in the aetiopathogenesis of either nodular or superficial BCC. The presence of HPV appears to be more related to actinic damage and possibly to an alteration of the barrier function associated with ageing. © 2010 The Authors Journal of the European Academy of Dermatology and Venereology © 2010 European Academy of Dermatology and Venereology.

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