Search

Objectives: This study is aimed at analyzing the impact of the main factors contributing to short and long-term mortality in patients at final stages of heart failure (HF). Setting: Patients attended at any of the 279 primary health care centers belonging to the Institut Català de la Salut, in Catalonia (Spain). Participants: Patients with Advanced HF. Design: Multicenter cohort study including 1148 HF patients followed for one-year after reaching New York Heart Association (NYHA) IV. Main measurements: The primary outcome was all-cause mortality. Multivariate logistic regression models were performed to assess the outcomes at 1, 3, 6, and 12 months. Results: Mean age of patients was 82 (SD 9) years and women represented 61.7%. A total of 135 (11.8%) and 397 (34.6%) patients died three months and one year after inclusion, respectively. Male gender, age, and decreased body mass index were associated with higher mortality at three, six and twelve months. In addition, low systolic blood pressure levels, severe reduction in glomerular filtration, malignancy, and higher doses of loop diuretics were related to higher mortality from 6 to 12 months.The most important risk factor over the whole period was presenting a body mass index lower than 20 kg/m2 (three months OR 3.06, 95% CI: 1.58–5.92; six months OR 4.42, 95% CI: 2.08–9.38; and 12 months OR 3.68, 95% CI: 1.76–7.69). Conclusions: We may conclude that male, age, and decreased body mass index determined higher short-term mortality in NYHA IV. In addition, low systolic blood pressure, reduced glomerular filtration, malignancy, and higher doses of loop diuretics contribute to increasing the risk of mortality at medium and long-term. Such variables are easily measurable and can help to decide the best way to face the most advances stages of the disease. Resumen: Objetivos: Analizar los factores que contribuyen a la mortalidad de pacientes en las etapas finales de la insuficiencia cardiaca (IC). Ámbito: Centros de atención primaria del Institut Català de la Salut, Cataluña, España. Participantes: Pacientes con IC avanzada. Diseño: Estudio de cohortes multicéntrico. Incluyó 1.148 pacientes de IC seguidos durante un año tras el registro de estadio funcional NYHA IV. Mediciones principales: El resultado principal fue la mortalidad por todas las causas. Se realizaron modelos de regresión logística multivariada (1, 3, 6 y 12 meses). Resultados: Edad media 82 años (DE 9), las mujeres representaron el 61,7%. Un total de 135 (11,8%) y 397 (34,6%) pacientes murieron 3 meses y un año después de su inclusión. El sexo masculino, la edad y el índice de masa corporal (IMC)

Objetivo: Analizar el coste-efectividad de la aplicación de algoritmos diagnósticos en pacientes con sospecha de un primer episodio de trombosis venosa profunda (TVP) en la atención primaria, en comparación con la derivación sistemática a centros especializados. Diseño: Estudio observacional, transversal y analítico. Emplazamiento: Pacientes seleccionados en urgencias hospitalarias derivados desde la atención primaria para completar estudio y diagnóstico. Participantes: Se reclutó a 138 candidatos con clínica compatible con un primer episodio de TVP; 22 fueron excluidos (sin informe de derivación, clínica de más de 30 días, anticoagulados y TVP previa), incluyéndose finalmente a 116 pacientes, un 61% mujeres, de 71 años edad media. Mediciones principales: Variables de las escalas de probabilidad clínica de Wells y Oudega, dímero-D (portátil y hospitalario), ecografía-Doppler y costes directos generados por los 3 circuitos analizados: derivación sistemática de todos los pacientes, derivación según escala de Oudega o de Wells. Resultados: En el 18,9% se confirmó el diagnóstico de TVP. Las 2 escalas de probabilidad clínica presentaron una sensibilidad del 100% (IC del 95%: 85,1-100) y una especificidad alrededor del 40%. Con la aplicación de las escalas, se hubiesen podido evitar con total seguridad un tercio de las derivaciones a urgencias hospitalarias (p < 0,001) y se hubieran podido disminuir los costes del proceso diagnóstico en 8.620 € según Oudega y 9.741 € según Wells, por cada 100 pacientes atendidos. Conclusiones: La aplicación de algoritmos diagnósticos en las sospechas de TVP permitiría al médico de atención primaria una orientación más resolutiva y coste-efectiva del proceso diagnóstico.

Background Intensive interventions on smoking cessation increase abstinence rates. However, few electronic mail (E-mail) based intensive interventions have been tested in smokers and none in primary care (PC) setting. The aim of the present study is to evaluate the effectiveness of an intensive E-mail based intervention in smokers attending PC services. Methods/design Randomized Controlled Multicentric Trial. Study population: 1060 smokers aged between 18–70 years from Catalonia, Salamanca and Aragón (Spain) who have and check regularly an E-mail account. Patients will be randomly assigned to control or intervention group. Intervention: Six phase intensive intervention with two face to face interviews and four automatically created and personal E-mail patients tracking, if needed other E-mail contacts will be made. Control group will receive a brief advice on smoking cessation. Outcome measures: Will be measured at 6 and 12 months after intervention: self reported continuous abstinence (confirmed by cooximetry), point prevalence abstinence, tobacco consumption, evolution of stage according to Prochaska and DiClemente's Stages of Change Model, length of visit, costs for the patient to access Primary Care Center. Statistical analysis: Descriptive and logistic and Poisson regression analysis under the intention to treat basis using SPSS v.17. Discussion The proposed intervention is an E-mail based intensive intervention in smokers attending primary care. Positive results could be useful to demonstrate a higher percentage of short and long-term abstinence among smokers attended in PC in Spain who regularly use E-mail. Furthermore, this intervention could be helpful in all health services to help smokers to quit. Trial Registration Clinical Trials.gov Identifier: NCT01494246.

11 p.-2 fig.-3 tab., 11α‐hydroxylated steroid synthons are one of the most important commercially pharmaceutical intermediates used for the production of contraceptive drugs and glucocorticoids. These compounds are currently produced by biotransformation using fungal strains in two sequential fermentation steps. In this work, we have developed by a rational design new recombinant bacteria able to produce 11α‐hydroxylated synthons in a single fermentation step using cholesterol (CHO) or phytosterols (PHYTO) as feedstock. We have designed a synthetic operon expressing the 11α‐hydroxylating enzymes from the fungus Rhizopus oryzae that was cloned into engineered mutant strains of Mycolicibacterium smegmatis that were previously created to produce 4‐androstene‐3,17‐dione (AD), 1,4‐androstadiene‐3,17‐dione (ADD) from sterols. The introduction of the fungal synthetic operon in these modified bacterial chassis has allowed producing for the first time 11αOH‐AD and 11αOH‐ADD with high yields directly from sterols in a single fermentation step. Remarkably, the enzymes of sterol catabolic pathway from M. smegmatis recognized the 11α‐hydroxylated intermediates as alternative substrates and were able to efficiently funnel sterols to the desired hydroxylated end‐products., This work was supported by grants from the Ministry of Science and Innovation (BFU2006-15214-C03-01, BFU2009-11545-C03-03) and Ministry of Economy and Competitiveness (BIO2012-39695-C02-01).

The enzymatic lignocellulosic biomass conversion into value-added products requires the use of enzyme-rich cocktails, including β-glucosidases that hydrolyze cellobiose and cellooligosaccharides to glucose. During hydrolysis occurs accumulation of monomers causing inhibition of some enzymes; thus, glucose/xylose tolerant β-glucosidases could overcome this drawback. The search of new tolerant enzymes showing additional properties,such as high activity, wide-pH range, and thermal stability is very relevant to improve the bioprocess. We describe a novel β-glucosidase GH1 from the thermophilic Anoxybacillus thermarum (BgAt), which stood out by the robustness combination of great glucose/xylose tolerance, thermal stability, and high Vmax. The recombinant his-tagged-BgAt was overexpressed in Escherichia coli, was purified in one step, showed a high glucose/xylose tolerance, and activity stimulation (presence of 0.4M glucose/1.0M xylose). The optimal activity was at 65 °C - pH 7.0. BgAt presented an extraordinary temperature stability (48 h – 50 °C), and pH stability (5.5–8.0). The novel enzyme showed outstanding Vmax values compared to other β-glucosidases. Using p-nitrophenyl-β-D-glucopyranoside as substrate the values were Vmax (7614 U/mg), and KM (0.360 mM). These values suffer a displacement in Vmax to 14,026 U/mg (glucose), 14,886 U/mg (xylose), and KM 0.877mM (glucose), and 1.410mM (xylose)., This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, process no 2018/07522-6) and a scholarship to Tassio B. Oliveira (grant 2017/09000-4). MLTMP is a Research Fellow of Conselho de Desenvolvimento Científico e Tecnológico (CNPq, process 301963/2017-7). The project also received grants from National Institute of Science and Technology of Bioethanol,INCT, CNPq 465319/2014-9/FAPESP nº 2014/50884-5). PZA was a fellow from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) funding code 001 and Programa de Doutorado Sanduiche no Exterior, PDSE no 88881.135684/2016-01.

5 p.-3 fig., Pederin is a potent polyketide toxin that causes severe skin lesions in humans after contact with insects of genus Paederus. Due to its potent anticancer activities, pederin family compounds have raised the interest of pharmaceutical industry. Despite the extensive studies on the cluster of biosynthetic genes responsible for the production of pederin, it has not yet been possible to isolate and cultivate its bacterial endosymbiont producer. However, the marine bacterium Labrenzia sp. PHM005 was recently reported to produce labrenzin, the closest pederin analog. By cloning a synthetic pedO gene encoding one of the three O-methyltraferase of the pederin cluster into Labrenzia sp. PHM005 we have been able to produce pederin for the first time by fermentation in the new recombinant strain., The present study was funded by the Ministry of Economy and Competitiveness of Spain under the program Retos/Colaboración with the project number RTC-2016-4892-1 (DESPOL)., Biotecnología aplicada al Desilenciamiento génico de Policétidos y Péptidos no ribosomales bioactivos en bacterias marinas. DESPOL

20 p.-12 fig.-1 tab., In this work, Caenibius tardaugens NBRC 16725 (strain ARI-1) (formerly Novosphingobium tardaugens) was isolated due to its capacity to mineralize estrogenic endocrine disruptors. Its genome encodes the edc genes cluster responsible for the degradation of 17β-estradiol, consisting of two putative operons (OpA and OpB) encoding the enzymes of the upper degradation pathway. Inside the edc cluster, we identified the edcR gene encoding a TetR-like protein. Genetic studies carried out with C. tardaugens mutants demonstrated that EdcR represses the promoters that control the expression of the two operons. These genetic analyses have also shown that 17β-estradiol and estrone, the second intermediate of the degradation pathway, are the true effectors of EdcR. This regulatory system has been heterologously expressed in Escherichia coli, foreseeing its use to detect estrogens in environmental samples. Genome comparisons have identified a similar regulatory system in the edc cluster of Altererythrobacter estronivorus MHB5, suggesting that this regulatory arrangement has been horizontally transferred to other bacteria., This research was funded by the Ramón Areces Foundation.

9 p.-7 fig., Hypertension is a multifactorial disorder considered one of the major causes of premature death worldwide. This pathology is associated with vascular functional/structural alterations in which nitric oxide (NO) and oxygen reactive species participate. On the other hand, the use of microalgae extracts in the treatment of cardiovascular diseases is increasing. Based on the antioxidant and antihypertensive properties of Spirulina, this study aims to investigate the effect of an aqueous extract of Spirulina on the vasodilator function of the aorta from spontaneously hypertensive rats (SHR), analyzing the functional role of NO. For this, aortic segments from male SHR were divided into two groups, one control and the other exposed to an Spirulina aqueous extract (0.1% w/v, for 3 hours), to analyze (i) the production of NO, superoxide anion, and hydrogen peroxide; (ii) the vasodilator response induced by acetylcholine (ACh), by the NO donor and sodium nitroprusside (SNP), and by the KATP channel opener and pinacidil; and (iii) the expression of the p-Akt, p-eNOS, and HO-1 proteins. The results showed that the aqueous Spirulina extract (i) increased the production of NO, did not significantly modify that of superoxide, while decreased that of hydrogen peroxide; (ii) increased the vasodilatory responses induced by ACh, NPS, and pinacidil; and (iii) increased the expression of p-Akt and HO-1. These results suggest that incubation with the aqueous Spirulina extract improves the vascular function of arteries from SHR by increasing the release/bioavailability/function of NO. Increased KATP channel activation and expression of pAkt and HO-1 appear to be participating in these actions., This study was supported by grants from Comunidad de Madrid (S2013/ABI-2783, “INSPIRΑ1-CM”) and Fondo Europeo de Desarrollo Regional to JLG, CO, and MF; the Spanish Ministerio de Economía, Industria y Competitividad (CTQ2017-86170-R) to CO; and by the Fondo de Investigaciones Sanitarias (PI19/01282) to MF.

21 p.-14 fig.-2 tab., We have analyzed the catabolism of estrogens in Novosphingobium tardaugens NBRC 16725, which is able to use endocrine disruptors such as 17β-estradiol, estrone, and estriol as sole carbon and energy sources. A transcriptomic analysis enabled the identification of a cluster of catabolic genes (edc cluster) organized in two divergent operons that are involved in estrogen degradation. We have developed genetic tools for this estrogen-degrading bacterium, allowing us to delete by site-directed mutagenesis some of the genes of the edc cluster and complement them by using expression plasmids to better characterize their precise role in the estrogen catabolism. Based on these results, a catabolic pathway is proposed. The first enzyme of the pathway (17β-hydroxysteroid dehydrogenase) used to transform 17β-estradiol into estrone is encoded out of the cluster. A CYP450 encoded by the edcA gene performs the second metabolic step, i.e., the 4-hydroxylation of estrone in this strain. The edcB gene encodes a 4-hydroxyestrone-4,5-dioxygenase that opens ring A after 4-hydroxylation. The initial steps of the catabolism of estrogens and cholate proceed through different pathways. However, the degradation of estrogens converges with the degradation of testosterone in the final steps of the lower catabolic pathway used to degrade the common intermediate 3aα-H-4α(3′-propanoate)7a-β-methylhexahydro-1,5-indanedione (HIP). The TonB-dependent receptor protein EdcT appears to be involved in estrogen uptake, being the first time that this kind of proteins has been involved in steroid transport., This work was supported by the Ramon Areces Foundation (ELISA project).

10 p.-5 fig.-2 tab., Motivation Synthetic microbial communities begin to be considered as promising multicellular biocatalysts having a large potential to replace engineered single strains in biotechnology applications, in pharmaceutical, chemical and living architecture sectors. In contrast to single strain engineering, the effective and high-throughput analysis and engineering of microbial consortia face the lack of knowledge, tools and well-defined workflows. This manuscript contributes to fill this important gap with a framework, called FLYCOP (FLexible sYnthetic Consortium OPtimization), which contributes to microbial consortia modeling and engineering, while improving the knowledge about how these communities work. FLYCOP selects the best consortium configuration to optimize a given goal, among multiple and diverse configurations, in a flexible way, taking temporal changes in metabolite concentrations into account., Results In contrast to previous systems optimizing microbial consortia, FLYCOP has novel characteristics to face up to new problems, to represent additional features and to analyze events influencing the consortia behavior. In this manuscript, FLYCOP optimizes a Synechococcus elongatus-Pseudomonas putida consortium to produce the maximum amount of bio-plastic (PHA, polyhydroxyalkanoate), and highlights the influence of metabolites exchange dynamics in a four auxotrophic Escherichia coli consortium with parallel growth. FLYCOP can also provide an explanation about biological evolution driving evolutionary engineering endeavors by describing why and how heterogeneous populations emerge from monoclonal ones., Availability and implementation Code reproducing the study cases described in this manuscript are available on-line: https://github.com/beatrizgj/FLYCOP, Supplementary information Supplementary data are available at Bioinformatics online., Issue Section: SYSTEMS, This work was supported from the European Union's Horizon 2020 Research and Innovation Programme under Grant Agreement no 686585, and the Spanish Ministry of Economy and Competitivity through the RobDcode grant (BIO2014-59528-JIN).

13 p.-5 fig.-1 tab., 14α-hydroxylated steroids are starting materials for the synthesis of contraceptive and anti-inflammatory compounds in the steroid industry. A synthetic bacterial operon containing the cytochrome P450 CYP103168 and the reductase CPR64795 of the fungus Cochlioboluslunatus able to hydroxylate steroids has been engineered into a shuttle plasmid named pMVFAN. This plasmid was used to transform two mutants of Mycolicibacterium smegmatis named MS6039-5941 and MS6039 that accumulate 4-androstene-3,17-dione (AD), and 1,4-androstadiene-3,17-dione (ADD), respectively. The recombinant mutants MS6039-5941 (pMVFAN) and MS6039 (pMVFAN) were able to efficiently express the hydroxylating CYP system of C.lunatus and produced in high yields 14αOH-AD and 14αOH-ADD, respectively, directly from cholesterol and phytosterols in a single fermentation step. These results open a new avenue for producing at industrial scale these and other hydroxylated steroidal synthons by transforming with this synthetic operon other Mycolicibacterium strains currently used for the commercial production of steroidal synthons from phytosterols as feedstock., This work was supported by grants from the Ministry of Science and Innovation (BFU2006-15214-C03-01, BFU2009-11545-C03-03) and Ministry of Economy and Competitiveness (BIO2012-39695-C02-01).

In this work we report the complete sequence and assembly of the estradiol-degrading bacterium Novosphingobium tardaugens NBRC 16725 genome into a single contig using the Pacific Biosciences RS II system., We acknowledge the financial support provided by the Ramón Areces Foundation.

14 p.-6 fig.-1 tab., Escherichia coli have developed one of the most efficient regulatory response mechanisms to phosphate starvation. The machinery involves a cascade with a two-component system (TCS) that relays the external signal to the genetic circuit, resulting in a feedback response. Achieving a quantitative understanding of this system has implications in synthetic biology and biotechnology, for example, in applications for wastewater treatment. To this aim, we present a computational model and experimental results with a detailed description of the TCS, consisting of PhoR and PhoB, together with the mechanisms of gene expression. The model is parameterised within the feasible range, and fitted to the dynamic response of our experimental data on PhoB as well as PhoA, the product of this network that is used in alkaline phosphatase production. Deterministic and stochastic simulations with our model predict the regulation dynamics in higher external phosphate concentrations while reproducing the experimental observations. In a cycle of simulations and experimental verification, our model predicts and explores phenotypes with various synthetic promoter designs that can optimise the inorganic phosphate intake in E. coli. Sensitivity analysis demonstrates that the Pho-controlled genes have a significant influence over the phosphate response. Together with experimental findings, our model should thus provide insights for the investigations on engineering new sensors and regulators for living technologies., This work has been partially funded by the European Union’s Horizon 2020 research and innovation programme under the grant agreement No 686585 - LIAR, Living Architecture.

7p.-2 fig.-4 tab., Several recent studies have shown that Arthrospira sp. supplementation of feeds has a beneficial effect on fish health and growth, but less is known about its possible effects on stress responsiveness. The present study was designed to evaluate using Arthrospira platensis as a feed supplement for Oreochromis niloticus fry, reared in recirculating aquaculture systems. Two isocaloric and isonitrogenous fishmeal-based diets were prepared with 0% and 1% A. platensis and fed to fry, approximately 10 mg live weight at the beginning of the experimental period (n=16 tanks, 8 tanks per treatment), at a feeding rate of 6% live weight, four meals a day for 50 d. The weight gain per tank, specific growth rate and feed conversion ratio were similar among treatments but A. platensis supplementation significantly increased survival (p, Cofunded by the Community of Madrid with structural funds from the European Union FEDER (INSPIRA1, Ref. S2013/ ABI2783).

7 p.-1 fig.-2 tab., The genome of MM1 (40,248 bp), a temperate bacteriophage from the Spain(23F)-1 multiresistant epidemic clone of Streptococcus pneumoniae, is organized in 53 open reading frames (ORFs) and in at least five functional clusters. Bioinformatic and N-terminal amino acid sequence analyses enabled the assignment of possible functions to 26 ORFs. Analyses comparing the MM1 genome with those of other bacteriophages revealed similarities, mainly with genomes of phages infecting gram-positive bacteria, which suggest recent exchange of genes between species colonizing the same habitat., This work was supported by grants from the Dirección General de Investigación Científica y Técnica (BCM2000-1002) and from Programa de Grupos Estratégicos de la Comunidad Autónoma de Madrid.

8 p.-6 fig.-1 tab., The nucleotide sequences of the hpaB and hpaC genes encoding the 4-hydroxyphenylacetate 3-hydroxylase from Escherichia coli W ATCC 11105 have been determined. These genes appear to be part of an operon and encode two proteins of 58,781 and 18,679 Da, respectively, that are required for hydroxylase activity. This aromatic hydroxylase is NADH-dependent and uses FAD as the redox chromophore. The largest component (HpaB) has been purified by affinity chromatography in Cibacron blue. E. coli cells that express exclusively hpaB showed only a very low hydroxylase activity that was enhanced in the presence of extracts containing the smallest protein HpaC. This behavior resembles that of the coupling protein of the 4-hydroxyphenylacetate 3-hydroxylase from Pseudomonas putida, and it might prevent the wasteful oxidation of NADH in the absence of substrate. Using a promoter-probe plasmid we have demonstrated that the hpaBC operon is expressed by a promoter inducible by 4-hydroxyphenylacetic acid. A gene, named hpaA, encoding a protein homologous to the XylS/AraC family of regulators, was identified upstream of the hydroxylase operon. The role played by HpaA in the regulation of the hpaBC operon remains to be elucidated. Since HpaB is not homologous to other aromatic hydroxylases, we suggest that the E. coli 4-hydroxyphenylacetate 3-hydroxylase is the first member of a new family of two-component aromatic hydroxylases sequenced so far., This work was supported by Grant AMB94-1038-C02-02 from the Comisión Interministerial de Ciencia y Tecnologia.

Introducción. La evaluación y manejo de la presión arterial (PA) es importante en la fase aguda del ictus. La monitorización ambulatoria de la PA (MAPA) se utiliza en el estudio y evaluación de la hipertensión arterial (HTA), pero apenas se ha estudiado en la fase aguda del ictus. Este estudio pretende investigar la PA obtenida por Monitorización de la PA durante 24 horas (MPA24h) el primer día del ictus, utilizando los equipos de la MAPA, analizando sus posibles diferencias según el tipo de ictus y su valor pronóstico precoz. Material y Métodos. Estudio observacional, longitudinal, de cohortes prospectivo sobre ictus de menos de 24 horas de evolución ingresados en el Hospital General La Mancha Centro de Alcázar de San Juan durante 4 años a los que se realizó MPA24h el primer día de ingreso y se determinó la escala de Rankin modificada (mRS) al séptimo día. Se ha realizado regresión lineal y logística binaria multivariante utilizando como variable de control del sesgo de confusión el índice de propensión (IP) obtenido como las probabilidades de respuesta positiva en un modelo de regresión multinominal con la variable dependiente politómica tipo de ictus y la variables independientes predictoras, sexo, HTA, diabetes mellitus (DM), dislipemia, insuficiencia cardiaca (IC), puntuación National Institute of Health Stroke Scale (NIHSS) al ingreso, enfermedad intercurrente, edad, ictus previo y tratamiento antiagregante. Resultados. Se han reclutado 499 ictus, con una mediana de edad de 74 años (amplitud intercuartil –AIC– de 67 a 79 años), 282 (56,5%) son varones, 297 (59,5%) hipertensos y 183 (36,7%) diabéticos. El 20,0% (100 pacientes) habían tenido un ictus previo, 25 (5%) un IAM previo y 68 (13,6%) tenían fibrilación auricular (FA). La mediana de puntuación de la NIHSS al ingreso es de 6 (AIC 3 a 13) puntos. La distribución por tipos de ictus es de 197 (37,5%) ictus lacunares, 99 (19,8%) cardioembólicos, 155 (31,1%) aterotrombóticos y 58 (11,6%) hemorrágicos. Se han encontrado 392 (78,9%) casos de buen pronóstico a los 7 días con mRS 0-3 y 105 (21,1%) casos de mal pronóstico con mRS 4-7. Las medianas de PA sistólica (PAS) son 134 (AIC 119 a 147) mmHg para ictus lacunar, 138 (AIC 124 a 156) mmHg para cardioembólico, 145 (AIC 127 a 160) mmHg para aterotrombótico y 144 (AIC 129 a 161) mmHg para ictus hemorrágico. Las medianas de PA diastólica (PAD) son 71 (AIC 64 a 78,5) mmHg en ictus lacunar, 70 (AIC 62 a 78) mmHg en cardioembólico, 74 (AIC 67 a 84) mmHg en aterotrombótico y 80 (AIC 70 a 93) en hemorrágico. En el análisis multivariante ajustado por el IP, no se observan diferencias significativas en los valores de PAS (p>0,200). Las diferencias significativas se observan en los valores de PAD (p=0,006), con diferencias significativas en las comparaciones múltiples de PAD entre hemorrágico-lacunar (diferencia de medias –DMed– 4,5 mmHg; intervalo de confianza –IC– 95% 0,5 a 8,4 mmHg; p=0,028), hemorrágico-cardioembólico (DMed 7,5 mmHg; IC 95% 3,3 a 11,6 mmHg; p0,200) ni de PAD (p>0,200) ni en los patrones circadianos de PAS (p>0,200), siendo diferentes los patrones circadianos de PAD (p=0,001). Las medianas de PAS y de PAD en el grupo de mal pronóstico son 148 (AIC 132 a 168) mmHg y 74 (AIC 65 a 83,5) mmHg, respectivamente, y en el grupo de buen pronóstico son 136 (AIC 21 a 150) mmHg y 73 (AIC 65 a 81) mmHg, respectivamente. En el análisis con regresión logística binaria multivariante ajustado por el IP, no se asocian a mal pronóstico los valores de PAS (razón de odds –RO– 1,01; IC 95% 1,00 a 1,02; p=0,111) ni de PAD (RO 1,00; IC 95% 0,98 a 1,02; p>0,200) del día 1. Se encuentra relación con el pronóstico en los cocientes nocturno/diurno de PAS (multiplicado por 100: RO 1,05; IC 95% 1,01 a 1,09; p=0,022) y de PAD (multiplicado por 100: RO 1,04; IC 95% 1,00 a 1,07; p=0,045). También tiene relación con el pronóstico la caída el día 7 respecto al día 1 tanto de la PAS (RO 0,97; IC 95% 0,95 a 0,99; p=0,002) como de la PAD (RO 0,93; IC 95% 0,90 a 0,97; p

Los pacientes que presentan una enfermedad crítica no deberían ser entendidos solamente como enfermos que son tratados en la UCI, pues la enfermedad debe considerarse como una trayectoria que comienza con el deterioro agudo y termina cuando el paciente ha recobrado un estado salud y una situación funcional que pueden ser considerados adecuados y estables. Así, la enfermedad del paciente crítico debe ser considerado un proceso largo que comprende su estancia en UCI, su posterior estancia en la sala de hospitalización convencional y su evolución tras el alta del hospital (89). En la UCI es importante valorar la capacidad funcional y los síntomas clínicos del paciente. En primer lugar debe definirse el impacto de la capacidad funcional del paciente previo a su ingreso en UCI sobre los resultados a largo plazo y las medidas terapéuticas aplicadas (136). En segundo lugar, sería importante conocer o estimar qué capacidad funcional y clínica residual tendría el paciente tras su alta de la UCI (137-140). La predicción de los resultados es importante no solo para los profesionales de la salud involucrados en el cuidado del paciente, lo que permitiría seleccionar mejor los tratamientos aplicados, sino también y de forma especial para sus familias y el entorno social, faltando el diseño de programas de cuidados posthospitalarios (141,142). En este sentido, debe señalarse que con frecuencia las familias deben realizar cambios sensibles en su estilo de vida para adaptarse a las nuevas circunstancias de los pacientes que han padecido una enfermedad crítica, que presentan en muchos casos un grado variable de dependencia (143). Como ya se ha comentado, dentro de los pacientes de UCI, el grupo con diagnóstico de SDMO es el que mayor estrés físico padece, siendo por ello la primera causa de fallecimiento en UCI (144). El fracaso de los principales órganos y sistemas representa una situación muy frecuente en los enfermos de UCI, y ello conduce con frecuencia a estancias prolongadas y a la necesidad de tratamientos complejos o agresivos (ventilación mecánica, drogas vasoactivas). A pesar de ello, son pocos los trabajos realizados sobre el pronóstico a largo plazo de los pacientes de UCI, y además la mayoría se han centrado en pacientes politraumatizados o con síndrome coronario agudo. En consecuencia, nos ha parecido de interés realizar un estudio en pacientes que ingresan en una UCI con un SDMO y evaluar estudiar la situación funcional y la sintomatología residual a largo plazo.

[ES]El Sarcoma de Ewing (SE) es un tumor maligno que afecta mayoritariamente a niños o adultos jóvenes. Dentro del grupo de sarcomas, después del osteosarcoma, es el que tiene peor pronóstico con una media de supervivencia del 30% en los casos metastásicos. A nivel molecular el SE se caracteriza por la presencia de una fusión génica entre el gen EWS y genes de la familia ETS principalmente FLI 1 (>90% casos) y ERG (5-10% casos . El producto de esta fusión es una proteína quimérica que actúa como factor de transcripción aberrante, desregulando ciertos genes tales como CD99 y genes relacionados con la ruta de IGF1R, entre otros genes. A nivel morfológico, las células de SE se caracterizan por la presencia de una elevada expresión del receptor de superficie CD99 en el 99% de las células. El tratamiento de primera línea se basa por regímenes multi-terapéuticos incluyendo quimioterapia con combinaciones de doxorubicina, vincristina, ifosfamida y etopósido, combinada con radioterapia y/o cirugía. Además en pacientes con tumores de mal pronóstico se han realizado varios ensayos clínicos con inhibidores de PARP, inhibidores de IGF1R y trasplante autólogo de células stem combinado con radioterapia, entre otros tratamientos. Todos estos estudios reflejan que la mejor estrategia terapéutica es el uso de de multi-tratamientos. En este sentido, estudios preclínicos, in vitro e in vivo han mostrado varias estrategias terapéuticas que podrían ser eficaces en el tratamiento de pacientes con SE de entre los cuales hay que destacar el bloqueo de las dianas clave en SE: la fusión génica, el eje IGF1-IGF1R y en segundo plano la sobre expresión de CD99., Este proyecto de tesis ha sido financiado por:- La Fundação para a Ciência e Tecnologia, Governo de Portugal, con el Proyecto: “The pathogenesis of Ewing Sarcoma: implications for novel treatment strategies” (SFRH/BD/69318/2010).- La Red de Excelencia de la Comunidad Europea EUROBONET, con el Proyecto Europeo “Molecular Pathology of Bone Tumors” (FP6-2004-Lifescihealth-5, 018814).- European Clinical trials in Rare Sarcomas within an integrated translational trial network (EUROSARC) FP7-HEALTH-2011-two-stage (Project ID 278742 Comisión Europea)- El Ministerio de Ciencia e Innovación, a través del Instituto de Salud Carlos III(RD12/0036/0017, RD06/0020/0059, PI1203102, PI1100018, PI081828, PI052524)-La Fundación Asociación Española Contra el Cáncer (AECC), con el proyecto “Búsqueda, validación y traslación clínica de nuevas dianas terapéuticas a partir de estudios de genómica y epigenómica integrativa en sarcoma de Ewing”. (GCB13131578DE Á)