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Resumen Introduccion La arteriosclerosis coronaria se ha relacionado con la deplecion y la alteracion funcional de las celulas progenitoras endoteliales (EPC). Nos propusimos determinar el numero de celulas CD34 + /KDR + y CD34 + /CD144 + en pacientes sometidos a revascularizacion miocardica quirurgica, comparados con pacientes operados de recambio valvular, asi como el efecto de su plasma sobre EPC cultivadas de donantes sanos. Metodos El numero de CD34 + /KDR + y CD34 + /CD144 + se determino por citometria de flujo. Las EPC cultivadas se obtuvieron de residuos leucoplaquetarios transfusionales. La apoptosis se midio por fragmentacion de ADN. La expresion de CD34 y CD144 se determino por Western blot. Resultados Encontramos un menor numero de CD34 + /CD144 + en pacientes revascularizados comparados con los de recambio valvular, pero en ambos casos fue un numero mayor que el hallado en sujetos sanos. El plasma de pacientes revascularizados tuvo un efecto antiapoptotico sobre EPC de sujetos sanos (al dia 7 de aislamiento), y este efecto fue inhibido por los bloqueadores de la via del TGF-beta1 SIS3 y SB-431542. Previo a ello, en el dia 4, el plasma de los pacientes revascularizados aumento la expresion in vitro de CD34 y CD1144. Conclusion El plasma de los pacientes revascularizados es antiapoptotico y aumenta la expresion de CD34 y CD144 en EPC de donantes sanos. Por lo tanto, el numero suboptimo de CD34 + /CD144 + que se observa in vivo sugiere un defecto en su liberacion de la medula osea.

Inflammation is a common feature in cardiovascular diseases, including diabetes mellitus. In addition to the well-known inflammatory role of cyclo-oxygenase-2 (COX-2), this protein has also been implicated in apoptosis resistance in tumour cells. Vascular smooth muscle cells (VSMC) from diabetic patients are also resistant to apoptosis because of an increased abundance of B cell lymphoma 2 protein (BCL2). In this work, we investigated whether overproduction of COX-2 was involved in the resistance to apoptosis in VSMC from diabetic patients. VSMC were obtained from internal mammary arteries from patients who had undergone coronary artery bypass graft surgery. Apoptosis was measured by DNA fragmentation, BCL2 degradation and cytochrome c release. Apoptosis induced by C-reactive protein in cells from non-diabetic patients was mediated by COX-2. VSMC from diabetic patients showed higher basal levels of COX-2 compared with those from non-diabetic patients. Transfection of VSMC from non-diabetic patients with a plasmid containing COX-2 (also known as PTGS2) increased basal production of COX-2 and BCL2 and mimicked the resistance to apoptosis that occurs in diabetic patients. We also found a significant correlation (R = 0.846, p = 0.016) between COX-2 and BCL2 production in arterial rings from diabetic patients measured by confocal microscopy. However, inhibition of COX-2 production by small interfering RNA proved unable to reverse BCL2 production in diabetic VSMC. These results suggest a link between inflammation (COX-2) and apoptosis resistance (BCL2) in the arteries of diabetic patients. This relationship is not causative and the common production of these two proteins may be co-regulated by shared regulatory elements in diabetes.

Resumen Introduccion La disfuncion endotelial es un importante mediador de la mayor morbimortalidad cardiovascular de los pacientes diabeticos. Las celulas progenitoras endoteliales (EPC) parecen poseer defectos funcionales en estos pacientes. La pioglitazona aumenta el numero y funcion de las EPC y disminuye la mortalidad cardiovascular en la poblacion diabetica. Objetivos Determinar la capacidad de adhesion de EPC cultivadas sobre arterias de pacientes diabeticos y no diabeticos, asi como evaluar los efectos de la glucosa y la pioglitazona sobre estos parametros. Material y metodos Las EPC se aislaron de celulas mononucleares a partir de la capa leucoplaquetaria de sangre de donantes y se cultivaron sobre fibronectina en medio microvascular. La adhesion se midio marcando las celulas con 111 In-oxina y pasandolas ex vivo sobre una camara de flujo con arterias mamarias internas remanentes de cirugia cardiaca. La expresion de la proteina CXCR-4 se determino por citometria de flujo. Resultados Se observo una mayor adhesion de las EPC sobre las arterias de pacientes diabeticos. Esta adhesion disminuyo al incubar las celulas con glucosa a 15 mM. La co-incubacion con pioglitazona 1 μM restauro la capacidad adhesiva de las EPC. La glucosa a 15 mM disminuyo la expresion de CXCR-4 en las EPC cultivadas, en cambio, la pioglitazona a 1 μM fue capaz de aumentarla. Conclusion Pese a que las arterias de pacientes diabeticos tienen una mayor capacidad de reclutar EPC, la hiperglucemia disminuye las propiedades adhesivas de estas celulas.

Introduction: To evaluate the ultra-structural changes of the chorioretinal vessels in hypercholesterolemic rabbits after treatment with statins. Methods: New Zealand rabbits were divided into four groups: Control (G0; n=10), and fed a standard diet for 8 months; Hypercholesterolemic (G1, n=8), were fed a 0.5% cholesterol-enriched diet for 8 months. Statins group (G2A y B, n=8), were each fed a 0.5% cholesterol-enriched diet for 8 months plus administration of fluvastatin sodium (G2A) or pravastatin sodium (G2B) at a dose of 2 mg/Kg/day. Eyes were processed by transmission electron microscopy. Results: G1 had a buildup of lipids at the suprachoroidea that compressed the vascular layers. G2 had a substantial decrease in the number of foam cells and lipids but more collagen fibres in the suprachoroidea and vascular layers. The lumen of the choroidal vessels was opened more in G2 in comparison with G1, which had a reduction in the capillary lumen to the point of collapse due to compression and hypertrophy of endothelial and vascular smooth muscle cells. The normal appearance of endothelial cells in G2 was in contrast with the endothelial necrosis observed in G1. In G2, the thickness of the Bruch membrane and the basal membrane of the choroidal and retinal vessels were reduced in comparison with G1. Conclusions: Treatment with statins reduces the build up of lipids and the number of macrophages in the choroid. Additionally, they preserve the endothelial cells and open the vascular lumens of choroidal vessels.

Selective feeding of the earthworm Hormogaster elisae was investigated by granulometric analysis, chemical fractionation and physical fractionation of the soil in which representatives were cultivated, and of their casts. H. elisae behaved as an endogeic species, mainly consuming soil from which it weakly but positively selected the organic fractions of greatest size (mainly free organic matter) and negatively selected the largest mineral fraction (coarse sand). No significant selection of intermediate size fractions was recorded. The 0-2 μm granulometric fraction (clays) was also selected; the results suggest that some of the organic components of this size fraction can also be used by the species.

We report the results of a laboratory study of the aestivation of Hormogaster elisae to determine the nature of the inactive period (diapause or quiescence) and to evaluate the influence of soil moisture, temperature, season, and earthworm body weight on the process. The results showed that specimens of H. elisae underwent facultative diapause—paradiapause—characterized by the construction of aestivation chambers in which the animal coils up and its activity decreases. Soil moisture appeared to be the most important environmental factor involved in the onset of aestivation. Temperature and time of year also had some influence, but earthworm body weight appeared to have none. Aestivating earthworms showed a decrease of 41.6% in mean body weight. Once replaced in soil with 20% moisture content, they took 6.4±3.1 d to exit their chambers and another 6.5±3.6 d to recover their initial body weight.

Summary This paper reports the composition and horizontal distribution of an earthworm community at El Molar (Madrid), and the underlying causes of its distribution. Five species of earthworms were found. The most abundant species were Hormogaster elisae, Allolobophora rosea and Allolobophora caliginosa trapezoides, which showed an aggregate-type distribution. Coarse sand, organic matter and nitrogen appear to be the factors with the greatest influence on H. elisae distribution, whereas porosity and aeration have influence on A. c. trapezoides, and clay, organic matter and nitrogen on the presence of A. rosea.

The gut load and gut transit time (GTT) of the endogeic earthworm Hormogaster elisae in laboratory cultures at 18 °C and 23 oC were studied. The GTT, 5.25 h ± 0.40 at 18 oC and 3.63 h ± 0.46 at 23 oC, was determined by staining the soil with alimentary colouring (tartrazine). The gut load was calculated with two methods: earthworm mass difference, before and after voiding the gut, and dry mass of the gut content. The gut load ranged between 168 and 261 mg wet mass g live earthworm mass –1 (mass difference method) or 137–196 mg dry mass g live earthworm mass –1 (dry mass method). With the obtained data a potential annual soil turnover for H. elisae was calculated: 211–470 kg wet soil mass kg live earthworm mass –1 year –1 (mass difference method) or 176–325 kg dry soil mass kg live earthworm mass –1 year –1 (dry mass method).

Transforming growth factor β (TGF-β1) is a pleiotropic cytokine with many and complex effects in cell and tissue physiology. This is made possible by a very complex and interwoven signaling system, whose regulation continues to be the focus of a growing line of research. This complex regulation translates to a key role in cardiovascular physiology, hemostasis, and the blood-vessel interface. In accordance with this, the TGF-β1 pathway appears to be deregulated in related disorders, such as atherosclerotic vascular disease and myeloproliferative syndromes. It is expected that the growing amount of experimental and clinical research will yield medical advances in the applications of knowledge of the TGF-β1 pathway to diagnosis and therapeutics.

Background Cardiovascular disease has been linked to endothelial progenitor cell (EPC) depletion and functional impairment in atherosclerosis and aortic stenosis. EPCs may play a pivotal role in vascular grafting. However, the EPC depletion in coronary artery bypass grafting (CABG) patients has not been compared to coronary artery disease-free valvular replacement patients with aortic stenosis. Methods We aimed to assess the basal number of CD34+/KDR+ and CD34+/CD144+ cells in CABG patients, compared to aortic stenosis valvular replacement patients. 100 patients (51 CABG and 49 valvular surgery ones) were included in the present study. All CABG or valvular patients had angiographic demonstration of the presence or the absence of coronary artery disease, respectively. Numbers of CD34+/KDR+ and CD34+/CD144+ were assessed by flow cytometry of pre-surgical blood samples. Results We found a lower number of CD34+/CD144+ cells in CABG patients compared to valvular patients (0.21 ± 0.03% vs. 0.47 ± 0.08%), and this difference remained statistically significant after the P was adjusted for multiple comparisons (P = 0.01428). Both groups had more EPCs than healthy controls. Conclusions Pre-surgical CD34+/CD144+ numbers are decreased in CABG patients, compared to valvular patients with absence of coronary disease., This work was supported by FISS (Health Research Fund, PI080920) and Red Tematica de Investigacion Cardiovascular RECAVA (RD06/0014/1007, RD06/0014/1027), both from the Instituto de Salud Carlos III (ISCIII). We are grateful to Fernando Ortego and all the nurses and doctors from the Cardiac Surgery service, Hospital Clinico San Carlos, for their technical assistance. We acknowledge support from Centro de Microscopía y Citometría (Universidad Complutense) and the Regional Blood Transfusion Centre, Madrid.

Dabigatran is an emerging oral anticoagulant which is a direct inhibitor of thrombin activity. It has been approved in the European Union and the United States of America for the prevention of thrombosis after major orthopedic surgery. It has also been approved by the American Food and Drug Administration and the European Medicines Agency for the prevention of stroke in chronic atrial fibrillation. Dabigatran provides a stable anticoagulation effect without any need to perform periodical laboratory controls. Of note, there is a growing amount of clinical evidence which shows its safety and efficacy. For these reasons, dabigatran may suppose a revolution in oral anticoagulation. However, two important limitations remain. First, it is contraindicated in patients with end-stage renal disease. Second, there is no evidence of the prevention of thrombosis in mechanical heart valves.

The earthworm community in El Molar (Madrid) is studied, and its distribution patterns and relation with some soil factors are described by using geostatistic and multivariate tools. Six species were found, Hormogaster elisae, Allolobophora rosea and Allolobophora caliginosa being the three most abundant ones. These speciesexhibited a clumped distribution. The most dominant species, H. elisae, was distributed in patches of an average size of 45m in spring and more than 100m inautumn. A. rosea was aggregated in patches of an average size of 22m and A. caliginosa formed patches of an average size of 38 m. There seemed to be a positive correlation between the abundance of H. elisae and the percentage of total and coarse sands, as well as a negative correlation with clay, nitrogen, carbon and coarse loams contents, opposite to what was observed for A. rosea.

Background Transforming growth factor beta (TGF-β1) is a pleiotropic cytokine, which is deregulated in atherosclerosis; however the role of age in this process is unknown. We aimed to assess whether TGF-β1 signaling is affected by age. Methods Vascular smooth muscle cells (VSMC) were obtained from patients undergoing abdominal surgery. Levels of TGF-β1 were measured by ELISA in sera from 169 patients undergoing coronary artery bypass grafting (CABG). The p27 expression was determined by Western blot from internal mammary arteries (IMA) obtained from CABG patients (n = 13). In VSMC from these patients undergoing abdominal surgery, secretion of TGF-β1 was determined by ELISA of cell-conditioned media. Results In VSMC from aged patients we observed a lower TGF-β1 secretion, measured as TGF-β1 concentration in cell conditioned medium (p p = 0.0195). Conclusions VSMC from aged patients showed a higher degree of cellular senescence and it was associated to a lower TGF-β1 secretion and signaling.