Bases moleculares que regulan la expresión de PECAM-1/CD31: caracterización estructural y funcional de su región promotora

Tesis inéditas de la Universidad Complutense de Madrid, Facultad de Farmacia, leída el 07-05-1997Pecam-1 es una molécula de adhesión implicada en la migración transendotelial que se expresa en células hematopoyéticas y endoteliales. Para comprender los mecanismos que regulan su expresión se ha aislado un clon genómico conteniendo 1555PB de la región 5 flanqueante de Pecam-1 y el primer exón del gen Pecam- 1. Esta región carece de caja tata y contiene secuencias consenso para SP1, EGR-1, ETS, HLH, GATA y AP-2, C/EBP. YY1, CCACC, LYF-1, heptamero, octamero, Hmg, elementos NFKB, SSRE, ARRE, glucocorticoides y una secuencia alu. Delecciones en dirección 5’ y 3’ han demostrado actividad promotora específica de tejido dentro de dos fragmentos contiguos: 0,22 KB NHEI/BG1II/PSTI siendo la actividad transcripcional del fragmento de 0,22 KB específica de linaje mieloide y la de 0,44KB BG1II/PST1 específica de células endoteliales. El mínimo fragmento con mayor respuesta a los esteres de forbol se ha identificado en el fragmento de 0,22 KB NHEI/BG1II y los factores de transcripción implicados son SP1 y EGR1.Fac. de FarmaciaTRUEunpu

Bases moleculares que regulan la expresión de PECAM-1/CD31: caracterización estructural y funcional de su región promotora

Pecam-1 es una molécula de adhesión implicada en la migración transendotelial que se expresa en células hematopoyéticas y endoteliales. Para comprender los mecanismos que regulan su expresión se ha aislado un clon genómico conteniendo 1555PB de la región 5 flanqueante de Pecam-1 y el primer exón del gen Pecam- 1. Esta región carece de caja tata y contiene secuencias consenso para SP1, EGR-1, ETS, HLH, GATA y AP-2, C/EBP. YY1, CCACC, LYF-1, heptamero, octamero, Hmg, elementos NFKB, SSRE, ARRE, glucocorticoides y una secuencia alu. Delecciones en dirección 5’ y 3’ han demostrado actividad promotora específica de tejido dentro de dos fragmentos contiguos: 0,22 KB NHEI/BG1II/PSTI siendo la actividad transcripcional del fragmento de 0,22 KB específica de linaje mieloide y la de 0,44KB BG1II/PST1 específica de células endoteliales. El mínimo fragmento con mayor respuesta a los esteres de forbol se ha identificado en el fragmento de 0,22 KB NHEI/BG1II y los factores de transcripción implicados son SP1 y EGR1

Effectiveness and safety of glimepiride and iDPP4, associated with metformin in second line pharmacotherapy of type 2 diabetes mellitus: systematic review and meta-analysis

Objective: Our review analyses the studies that have specifically compared the association iDPP4/metformin with glimepiride/metformin, both in second line pharmacotherapy of type 2 diabetes mellitus (DM2). Methods: Systematic literature review with a meta-analysis of clinical trials comparing glimepiride with any iDPP4, both used together with metformin as a second line treatment of DM2. The effectiveness variables used were as follows: %HbA1c variation, fasting plasma glucose variation, patients achieving the therapeutic objective of HbA1c <7%, treatment dropouts due to lack of effectiveness and rescue treatments needed. The safety variables included were as follows: weight variation at the end of treatment; presentation of any type of adverse event; presentation of serious adverse events; patients who experienced any type of hypoglycaemia; patients who experienced severe hypoglycaemia; treatments suspended due to adverse effects; and deaths for any reason. Results: Four studies met the inclusion criteria. The group treated with glimepiride showed better results in all effectiveness variables. Regarding safety variables, the main differences observed were in the greater number of cases with hypoglycaemia in the group treated with glimepiride, and the serious adverse events or treatment discontinuations due to these which occurred in slightly over 2% more cases in this group compared to the iDPP4 group. The remaining adverse events, including mortality, did not show any differences between both groups. The variation in the weight difference between groups (2.1 kg) is not considered clinically relevant. Conclusions: A greater effectiveness is seen in the glimepiride/metformin association, which should not be diminished by slight differences in adverse effects, with absence of severe hypoglycaemia in over 98% of patients under treatment. The association of glimepiride/metformin, both due to cost as well as effectiveness and safety, may be the preferential treatment for most DM2 patients, and it offers a potential advantage in refractory hyperglycemic populations, tolerant to treatment.S

Enhanced Cytotoxic Activity of Mitochondrial Mechanical Effectors in Human Lung Carcinoma H520 Cells: Pharmaceutical Implications for Cancer Therapy

Cancer cell mitochondria represent an attractive target for oncological treatment as they have unique hallmarks that differ from their healthy counterparts, as the presence of a stronger membrane potential that can be exploited to specifically accumulate cytotoxic cationic molecules. Here, we explore the selective cytotoxic effect of 10-N-nonyl acridine orange (NAO) on human lung carcinoma H520 cells and compare them with healthy human lung primary fibroblasts. NAO is a lipophilic and positively charged molecule that promotes mitochondrial membrane adhesion that eventually leads to apoptosis when incubated at high micromolar concentration. We found an enhanced cytotoxicity of NAO in H520 cancer cells. By means Fluorescence lifetime imaging microscopy (FLIM) we also confirmed the formation of H-dimeric aggregates originating from opposing adjacent membranes that interfere with the mitochondrial membrane structure. Based on our results, we suggest the mitochondrial membrane as a potential target in cancer therapy to mechanically control the cell proliferation of cancer cells

Targeting gene expression to endothelium in transgenic animals: A comparison of the human ICAM-2, PECAM-1 and endoglin promoters

It is highly likely that successful pig-to-human xenotransplantation of vascularized organs will require genetic modification of the donor pig, and in particular of donor vascular endothelium. Promoters are generally tested in transgenic mice before generating transgenic pigs. Several promoters have been used to drive endothelial cell-specific expression in mice but none have yet been tested in pigs. We compared the promoters of three human genes that are predominantly expressed in vascular endothelium: intercellular adhesion molecule 2 (ICAM-2), platelet endothelial cell adhesion molecule 1 (PECAM-1) and endoglin. Expression of human complement regulatory proteins (hCRPs), directed by each of the promoters in mice, was largely restricted to vascular endothelium and leukocyte subpopulations. However, expression from the PECAM-1 promoter was weak in liver and non-uniform in the small vessels of heart, kidney, and lung. Conversely, expression from the endoglin promoter was consistently strong in the small vessels of these organs but was absent in larger vessels. The ICAM-2 promoter, which produced strong and uniform endothelial expression in all organs examined, was therefore used to generate hCRP transgenic pigs. Leukocytes from 57 pigs containing at least one intact transgene were tested for transgene expression by flow cytometry. Forty-seven of these transgenic pigs were further analyzed by immunohistochemical staining of liver biopsies, and 18 by staining of heart and kidney sections. Only two of the pigs showed expression, which appeared to be restricted to vascular endothelium in heart and kidney but was markedly weaker than in transgenic mice produced with the same batch of DNA. Thus, in this case, promoter performance in mice and pigs was not equivalent. The weak expression driven by the human ICAM-2 promoter in pigs relative to mice suggests the need for additional regulatory elements to achieve species-specific gene expression in pigs.This work was supported in part by grants from Ministerio de Ciencia y Tecnologia and Comunidad de Madrid to CB.Peer Reviewe

Role of age and comorbidities in mortality of patients with infective endocarditis

[Purpose]: The aim of this study was to analyse the characteristics of patients with IE in three groups of age and to assess the ability of age and the Charlson Comorbidity Index (CCI) to predict mortality. [Methods]: Prospective cohort study of all patients with IE included in the GAMES Spanish database between 2008 and 2015.Patients were stratified into three age groups:<65 years,65 to 80 years,and ≥ 80 years.The area under the receiver-operating characteristic (AUROC) curve was calculated to quantify the diagnostic accuracy of the CCI to predict mortality risk. [Results]: A total of 3120 patients with IE (1327 < 65 years;1291 65-80 years;502 ≥ 80 years) were enrolled.Fever and heart failure were the most common presentations of IE, with no differences among age groups.Patients ≥80 years who underwent surgery were significantly lower compared with other age groups (14.3%,65 years; 20.5%,65-79 years; 31.3%,≥80 years). In-hospital mortality was lower in the <65-year group (20.3%,<65 years;30.1%,65-79 years;34.7%,≥80 years;p < 0.001) as well as 1-year mortality (3.2%, <65 years; 5.5%, 65-80 years;7.6%,≥80 years; p = 0.003).Independent predictors of mortality were age ≥ 80 years (hazard ratio [HR]:2.78;95% confidence interval [CI]:2.32–3.34), CCI ≥ 3 (HR:1.62; 95% CI:1.39–1.88),and non-performed surgery (HR:1.64;95% CI:11.16–1.58).When the three age groups were compared,the AUROC curve for CCI was significantly larger for patients aged <65 years(p < 0.001) for both in-hospital and 1-year mortality. [Conclusion]: There were no differences in the clinical presentation of IE between the groups. Age ≥ 80 years, high comorbidity (measured by CCI),and non-performance of surgery were independent predictors of mortality in patients with IE.CCI could help to identify those patients with IE and surgical indication who present a lower risk of in-hospital and 1-year mortality after surgery, especially in the <65-year group

Adherencia al tratamiento con Hormona de Crecimiento Recombinante en niños deficitarios: Control terapéutico e impacto económico–EPÍTOME Informe IPE 2013/70–

El presente documento es un epítome del Informe IPE 2013/70 de la Agencia de Evaluación de Tecnologías Sanitarias (AETS) del Instituto de Salud Carlos III, publicado con el título «Adherencia al tratamiento con Hormona de Crecimiento Recombinante en niños deficitarios: Control terapéutico e impacto económico». Dado el carácter de resumen de esta publicación, se han omitido numerosos pormenores que el lector interesado puede encontrar en la edición del documento completo accediendo a la dirección web del ISCIII (Publicaciones) o directamente en: http://gesdoc.isciii.es/gesdoccontroller?action=download &id=13/11/2013-e161b95e2

Enhanced Cytotoxic Activity of Mitochondrial Mechanical Effectors in Human Lung Carcinoma H520 Cells: Pharmaceutical Implications for Cancer Therapy

8 pags, 5 figsCancer cell mitochondria represent an attractive target for oncological treatment as they have unique hallmarks that differ from their healthy counterparts, as the presence of a stronger membrane potential that can be exploited to specifically accumulate cytotoxic cationic molecules. Here, we explore the selective cytotoxic effect of 10-N-nonyl acridine orange (NAO) on human lung carcinoma H520 cells and compare them with healthy human lung primary fibroblasts. NAO is a lipophilic and positively charged molecule that promotes mitochondrial membrane adhesion that eventually leads to apoptosis when incubated at high micromolar concentration. We found an enhanced cytotoxicity of NAO in H520 cancer cells. By means Fluorescence lifetime imaging microscopy (FLIM) we also confirmed the formation of H-dimeric aggregates originating fromopposing adjacent membranes that interfere with the mitochondrial membrane structure. Based on our results, we suggest the mitochondrial membrane as a potential target in cancer therapy to mechanically control the cell proliferation of cancer cells.This work was supported by the ERC Starting Grant MITOCHON (ERC-StG-2013-338133), ERC Proof of Concept mitozippers (ERC-PoC-2017-780440) and FIS2015-70339-C2-1-R from the Spanish Ministry of Economy MINECO (IL-M); and FIS2015-70339-C2-2-R (MPL and CG)

Enhanced Cytotoxic Activity of Mitochondrial Mechanical Effectors in Human Lung Carcinoma H520 Cells: Pharmaceutical Implications for Cancer Therapy

Cancer cell mitochondria represent an attractive target for oncological treatment as they have unique hallmarks that differ from their healthy counterparts, as the presence of a stronger membrane potential that can be exploited to specifically accumulate cytotoxic cationic molecules. Here, we explore the selective cytotoxic effect of 10-N-nonyl acridine orange (NAO) on human lung carcinoma H520 cells and compare them with healthy human lung primary fibroblasts. NAO is a lipophilic and positively charged molecule that promotes mitochondrial membrane adhesion that eventually leads to apoptosis when incubated at high micromolar concentration. We found an enhanced cytotoxicity of NAO in H520 cancer cells. By means Fluorescence lifetime imaging microscopy (FLIM) we also confirmed the formation of H-dimeric aggregates originating from opposing adjacent membranes that interfere with the mitochondrial membrane structure. Based on our results, we suggest the mitochondrial membrane as a potential target in cancer therapy to mechanically control the cell proliferation of cancer cells