Flavodoxin overexpression confers tolerance to oxidative stress in beneficial soil bacteria and improves survival in the presence of the herbicides paraquat and atrazine.

AIM: To determine whether expression of a cyanobacterial flavodoxin in soil bacteria of agronomic interest confers protection against the widely used herbicides paraquat and atrazine. METHODS AND RESULTS: The model bacterium Escherichia coli, the symbiotic nitrogen-fixing bacterium Ensifer meliloti and the plant growth-promoting rhizobacterium Pseudomonas fluorescens Aur6 were transformed with expression vectors containing the flavodoxin gene of Anabaena variabilis. Expression of the cyanobacterial protein was confirmed by Western blot. Bacterial tolerance to oxidative stress was tested in solid medium supplemented with hydrogen peroxide, paraquat or atrazine. In all three bacterial strains, flavodoxin expression enhanced tolerance to the oxidative stress provoked by hydrogen peroxide and by the reactive oxygen species-inducing herbicides, witnessed by the enhanced survival of the transformed bacteria in the presence of these oxidizing agents. CONCLUSIONS: Flavodoxin overexpression in beneficial soil bacteria confers tolerance to oxidative stress and improves their survival in the presence of the herbicides paraquat and atrazine. Flavodoxin could be considered as a general antioxidant resource to face oxidative challenges in different micro-organisms. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of plant growth-promoting rhizobacteria or nitrogen-fixing bacteria with enhanced tolerance to oxidative stress in contaminated soils is of significant agronomic interest. The enhanced tolerance of flavodoxin-expressing bacteria to atrazine and paraquat points to potential applications in herbicide-treated soils.

Glycaemic control in patients with type 2 diabetes switching from premixed insulin to long-acting basal insulin analogue plus oral antidiabetic drugs: an observational study.

AIM: To evaluate whether administration of long-acting basal insulin analogue plus oral antidiabetic drugs (OADs) improves glycaemic control in type 2 diabetic patients with glycosylated haemoglobin (HbA1c) > 7% (53 mmol/mol) under premixed insulin therapy. METHODS: This is a multicentre, observational, retrospective study performed in type 2 diabetic patients switching from premixed insulin to long-acting basal insulin analogue plus OADs. Data on patients' medical history and assessments were retrieved from patients' medical charts prior to switching the treatment and 6 months thereafter. RESULTS: A total of 131 evaluable patients were enrolled (mean age, 68.2 ± 9.4 years; female, 65.6%; mean diabetes duration, 12.7 ± 6.9 years; mean time on insulin therapy, 53.2 ± 41.9 months). Patients were receiving premixed insulin (once-daily, 4.7%; twice-daily, 85.0%; thrice-daily, 10.2%), 82.4% of whom in combination with OADs (metformin, 79.4%). After the treatment was switched, only 14.5% required intensification of treatment with additional preprandial insulin. HbA1c decreased -1.4% [mean ± SD, 8.4 ± 1.0% (68.7 ± 11.4 mmol/mol) vs. 7.0 ± 1.0% (53.6 ± 10.9 mmol/mol), p < 0.001] and the proportion of patients achieving HbA1c < 7% (53 mmol/mol) increased to 52.7% (p < 0.001). The percentage of patients with hypoglycaemia decreased (19.2% vs. 10.8%, p < 0.05; symptomatic, 17.6% vs. 4.6%, p < 0.01) and body weight diminished by -1.9 kg (mean ± SD, 78.5 ± 14.7 kg vs. 76.6 ± 13.9 kg, p < 0.05). Basal insulin plus OADs was considered more convenient and flexibly adapted to patients' life in 98.4% and 99.2% of patients, respectively. Additionally, 96.9% of patients reported being more satisfied and 96.9% would recommend it. CONCLUSIONS: Switching the treatment from premixed insulin to long-acting basal insulin analogue plus OADs is a feasible and convenient approach to improve glycaemic control of type 2 diabetic patients poorly controlled with premixed insulin under routine clinical practice conditions.

[Incidence of ischemic heart disease in the HERMEX study and its association with classical cardiovascular risk factors]. / Incidencia de cardiopatía isquémica en el estudio HERMEX y su asociación con los factores de riesgo cardiovascular clásicos.

OBJECTIVES: Knowledge about the incidence of coronary heart disease in our country is scarce and its sources are not comparable. Our aim was to determine the incidence of ischemic heart disease in a population cohort in a 7 years of follow-up, as well as the risk associated with the different classical cardiovascular risk factors. METHODS: Cohort study with a population sample of 2833 subjects, selected by the simple random method in a population between 25 and 79, from the Don Benito - Villanueva de la Serena (Badajoz) health area, response rate was 80.5%. All episodes of angina pectoris, lethal and non-lethal myocardial infarction were collected in individuals with no previous history of cardiovascular disease. Cumulative incidences and incidence rates adjusted per 100,000 person-years by sex and overall were calculated. RESULTS: From 2833 initial cohort participants, 103 were excluded due to a history of cardiovascular disease and 61 for losses, 2669 completed the follow-up (94.2%). The median follow-up was 6.9 years (IR 6.5-7.5), 56.4% women; 59 events were recorded. The overall incidence rate of ischemic heart disease was 327 cases/100,000 person-years, corresponding to 470 cases/100,000 for men and 211 cases/100,000 people-years for women. The classical cardiovascular risk factors that were associated with a higher risk of presenting events in the follow-up were dyslipidemia and arterial hypertension, in addition to male sex and age. CONCLUSIONS: The incidence of ischemic heart disease in the urban-rural population of Extremadura is high. The classical cardiovascular risk factors most strongly associated with events were dyslipidemia, and arterial hypertension.

Strength of association of classical vascular risk factors in young patients with ischaemic stroke: a case-control study.

INTRODUCTION: Recent studies have reported an increasing incidence of ischaemic stroke among young adults. However, the strength of the association between traditional vascular risk factors has not been fully established. METHODS: We compared 120 patients with a first ischaemic stroke before the age of 55 years admitted to the stroke unit of our centre with 600 healthy non-stroke controls from a population-based cohort study (HERMEX), matched for sex. Risk factors assessed included: hypertension, obesity, auricular fibrillation, current smoking, estimated glomerular filtration rate (eGFR), total cholesterol, low-density lipoprotein cholesterol (LDL-C), triglycerides, high-density lipoprotein cholesterol (HDL-C) and diabetes mellitus. We used logistic regression analysis and calculated population attributable risk. We performed an overall analysis, by sex and aetiological subgroup. RESULTS: Using logistic regression analysis, we found that overall, the significant risk factors were: hypertension (OR: 1.58; 95%CI: 1.01-2.50), atrial fibrillation (OR: 4.77; 95%CI: 1.20-19.00), low eGFR (OR: 4.74; 95%CI: 1.3-21.94) and low HDL-C (OR: 5.20; 95%CI: 3.29-8.21), as well as smoking for males (OR: 1.86; 95%CI: 1.14-3.03). LDL-C showed an inverse association with stroke. The population attributable risk for HDL-C was 37.8% and for hypertension 21.1%. In terms of aetiological subgroups, only low HDL-C was associated with stroke of undetermined aetiology. CONCLUSIONS: Hypertension, auricular fibrillation, low eGFR, and low HDL-C, plus tobacco use in men, are the main risk factors among patients under 55 years of age with a first ischaemic stroke. We believe that it would be of particular interest to further explore the management of low HDL-C levels as part of preventive strategies in young stroke patients.

Complications related to less-invasive haemodynamic monitoring.

BACKGROUND: The aim of this study was to evaluate the type and incidence of complications during insertion, maintenance, and withdrawal of central arterial catheters used for transpulmonary thermodilution haemodynamic monitoring (PiCCO™). METHODS: We conducted a prospective, observational, multicentre study in 14 European intensive care units (six countries). A total of 514 consecutive patients in whom haemodynamic monitoring by PiCCO™ was indicated were studied. RESULTS: Five hundred and fourteen PiCCO catheters (475 in femoral, 26 in radial, nine in axillary, and four in brachial arteries) were inserted. Arterial access was obtained on the first attempt in 86.4% of the patients. Minor problems such as oozing after insertion (3.3%) or removal of the catheter (3.5%) were observed, but no episodes of serious bleeding (more than 50 ml) were recorded. Small local haematomas were observed after insertion (4.5%) and after removal (1.2%) of the catheter. These complications were not more frequent in patients with coagulation abnormalities. The incidence of site inflammation and catheter-related infection was 2% and 0.78%, respectively. Other complications such as ischaemia (0.4%), pulse loss (0.4%), or femoral artery thrombosis (0.2%) were rare, transient, and all resolved with catheter removal or embolectomy, respectively. CONCLUSIONS: In this series of patients, central arterial catheters used for PiCCO™ monitoring were demonstrated to be a safe alternative for advanced haemodynamic monitoring.

[Development of animal experimental model for bacterial peritonitis]. / Desarrollo de modelo experimental animal de peritonitis bacteriana.

OBJECTIVE: The aim of the study was to develop a model of abdominal sepsis in the experimental animal. METHODS: Sprague-Dawley male rats of 5 weeks (N=39) were used. Initially, a pilot study (N = 9) was performed and distributed in 3 groups with 1cc inoculum of Escherichia coli ATCC 25922 intraperitoneally at concentrations of 10E8, 10E9 and 10E10 CFU. Subsequently, concentrations of 10E10 CFU are used in two groups of 3 rats with dilutions of 10 cc and 15 cc of distilled water respectively. Finally, a randomized trial of 24 rats was started in three treatment groups after intraperitoneal infection: Group I with physiological serum (N = 6), Group II with ceftriaxone (N = 9), Group III with ceftriaxone plus allicin (N = 9). Microbiological samples of blood and peritoneal fluid were made, as well as histopathological study of intraperitoneal organs (liver, diaphragm and peritoneum). RESULTS: Death of 100% of the rats infected with 10E10 E. coli UFC concentration with the dilution of 15 ml of distilled water and without antibiotic was oberved. The blood culture and peritoneal fluid culture was positive for the same strain in all of them. The formation of abscesses on the liver surface and polymorphonuclear infiltration in tissues were observed. CONCLUSIONS: The lethal dose of E. coli is 10E10 CFU diluted in 15 cc distilled water by intraperitoneal injection.

Patient characteristics, clinical course and factors associated to ICU mortality in critically ill patients infected with SARS-CoV-2 in Spain: A prospective, cohort, multicentre study. / Características, evolución clínica y factores asociados a la mortalidad en UCI de los pacientes críticos infectados por SARS-CoV-2 en España: estudio prospectivo, de cohorte y multicéntrico.

BACKGROUND: The clinical course of COVID-19 critically ill patients, during their admission in the intensive care unit (UCI), including medical and infectious complications and support therapies, as well as their association with in-ICU mortality has not been fully reported. OBJECTIVE: This study aimed to describe clinical characteristics and clinical course of ICU COVID-19 patients, and to determine risk factors for ICU mortality of COVID-19 patients. METHODS: Prospective, multicentre, cohort study that enrolled critically ill COVID-19 patients admitted into 30 ICUs from Spain and Andorra. Consecutive patients from March 12th to May 26th, 2020 were enrolled if they had died or were discharged from ICU during the study period. Demographics, symptoms, vital signs, laboratory markers, supportive therapies, pharmacological treatments, medical and infectious complications were reported and compared between deceased and discharged patients. RESULTS: A total of 663 patients were included. Overall ICU mortality was 31% (203 patients). At ICU admission non-survivors were more hypoxemic [SpO2 with non-rebreather mask, 90 (IQR 83 to 93) vs. 91 (IQR 87 to 94); P<.001] and with higher sequential organ failure assessment score [SOFA, 7 (IQR 5 to 9) vs. 4 (IQR 3 to 7); P<.001]. Complications were more frequent in non-survivors: acute respiratory distress syndrome (ARDS) (95% vs. 89%; P=.009), acute kidney injury (AKI) (58% vs. 24%; P<10-16), shock (42% vs. 14%; P<10-13), and arrhythmias (24% vs. 11%; P<10-4). Respiratory super-infection, bloodstream infection and septic shock were higher in non-survivors (33% vs. 25%; P=.03, 33% vs. 23%; P=.01 and 15% vs. 3%, P=10-7), respectively. The multivariable regression model showed that age was associated with mortality, with every year increasing risk-of-death by 1% (95%CI: 1 to 10, P=.014). Each 5-point increase in APACHE II independently predicted mortality [OR: 1.508 (1.081, 2.104), P=.015]. Patients with AKI [OR: 2.468 (1.628, 3.741), P<10-4)], cardiac arrest [OR: 11.099 (3.389, 36.353), P=.0001], and septic shock [OR: 3.224 (1.486, 6.994), P=.002] had an increased risk-of-death. CONCLUSIONS: Older COVID-19 patients with higher APACHE II scores on admission, those who developed AKI grades ii or iii and/or septic shock during ICU stay had an increased risk-of-death. ICU mortality was 31%.

Trends in stroke hospitalisation rates in Extremadura between 2002 and 2014: Changing the notion of stroke as a disease of the elderly. / Tendencias en las tasas de hospitalización por ictus en Extremadura en el periodo 2002-2014. Cambiando la idea del ictus como una enfermedad propia de la senectud.

BACKGROUND: The incidence of stroke in Spain has been evaluated in several studies, whose results are highly variable and not comparable. No studies of stroke have analysed epidemiological changes in younger patients. METHOD: We conducted a retrospective observational study using the Spanish health system's Minimum Data Set and included all patients older than 19 hospitalised due to stroke (ICD-9-CM codes 434.01, 434.11, 434.91, 430, 431, 432.9, 436, and 435) between 2002 and 2013. The analysis was performed using joinpoint regression. RESULTS: A total of 39,321 patients were identified (47.25% were women); 3.73% were aged 20-44, 6.29% were 45-54, 11.49% were 55-64, 23.89% were 65-74, and 54.60% were > 74 years. The hospitalisation rate due to ischaemic stroke has increased significantly in men aged 45-54 (+6.7%; 95% CI, 3.3-10.2) and in women aged 20-44 and 45-54 (+6.1%; 95% CI, 0.8-11.7 and +5.7%; 95% CI, 3.0-8.4, respectively). We also observed a significant increase in the rate of hospitalisation due to ischaemic stroke in men aged over 74 (+4.2%; 95% CI, 1.3-7.2). The rate of hospitalisations due to transient ischaemic attack has also increased significantly whereas the rate of hospitalisations due to brain haemorrhage has stabilised over time. CONCLUSIONS: Our results provide indirect evidence that the epidemiological profile of stroke is changing based on the increase in hospitalisation rates in young adults.

[Acute myocardial infarction complicating radiofrequency catheter ablation in a girl]. / Infarto agudo de miocardio tras ablación por radiofrecuencia de vía cardiaca anómala en un niño.

Paroxysmal supraventricular tachycardia is the most common tachyarrhythmia in childhood and adolescence. The current treatment of choice for managing and preventing frequent recurrences is radiofrequency catheter ablation, which provides a definitive solution in most cases. This technique is very effective but not devoid of potential complications, particularly in children and adolescents. We report the case of a 3-year-old girl who suffered acute myocardial infarction after occlusion of the right coronary artery during this procedure. We describe intraoperative and postoperative management. Progress was good and the patient was discharged 14 days after ablation.

Incidencia de cardiopatía isquémica en el estudio HERMEX y su asociación con los factores de riesgo cardiovascular clásicos / Incidence of ischemic heart disease in the HERMEX study and its association with classical cardiovascular risk factors

Objetivos: El conocimiento sobre la incidencia de enfermedad coronaria en nuestro país es escaso y sus fuentes poco comparables. Fue nuestro objetivo determinar la incidencia de cardiopatía isquémica en una cohorte poblacional con un seguimiento de 7 años y el riesgo asociado a los factores de riesgo cardiovascular clásicos. Métodos: Estudio de cohortes prospectivo en una muestra poblacional de 2833 sujetos, seleccionada por método aleatorio simple entre ciudadanos de 25 a 79 años del Área de Salud Don Benito – Villanueva de la Serena (Badajoz), tasa de respuesta 80,5%. Se recogieron todos los episodios de angina de pecho, infarto de miocardio letal y no letal en individuos sin antecedentes previos de enfermedad cardiovascular. Se calcularon las incidencias acumuladas y las tasas de incidencia ajustadas por 100.000 personas-año por sexo y global. Resultados: De los 2833 participantes iniciales, se excluyeron 103 por antecedentes de enfermedad cardiovascular y 61 por pérdidas, completando el seguimiento 2669 (94,2%). La mediana de seguimiento fue de 6,9 años (RI 6,5 - 7,5), 56,4% mujeres; se registraron 59 eventos. La tasa de incidencia global de cardiopatía isquémica fue 327 casos/100.000 personas-año correspondiendo 470 casos/100.000 al sexo masculino y 211 casos/100.000 personas-año al femenino. Los factores de riesgo cardiovascular clásicos que se asociaron con mayor riesgo de presentar eventos en el seguimiento fueron la dislipemia y la HTA, además del sexo masculino y la edad. Conclusiones: La incidencia de cardiopatía isquémica en la población urbano-rural de Extremadura, es elevada. Los factores de riesgo cardiovascular clásicos más fuertemente asociados a su aparición fueron la dislipemia, y la hipertensión arterial (AU)

Objectives: Knowledge about the incidence of coronary heart disease in our country is scarce and its sources are not comparable. Our aim was to determine the incidence of ischemic heart disease in a population cohort in a 7 years of follow-up, as well as the risk associated with the different classical cardiovascular risk factors. Methods: Cohort study with a population sample of 2833 subjects, selected by the simple random method in a population between 25 and 79, from the Don Benito – Villanueva de la Serena (Badajoz) health area, response rate was 80.5%. All episodes of angina pectoris, lethal and non-lethal myocardial infarction were collected in individuals with no previous history of cardiovascular disease. Cumulative incidences and incidence rates adjusted per 100,000 person-years by sex and overall were calculated. Results: From 2833 initial cohort participants, 103 were excluded due to a history of cardiovascular disease and 61 for losses, 2669 completed the follow-up (94.2%). The median follow-up was 6.9 years (IR 6.5–7.5), 56.4% women; 59 events were recorded. The overall incidence rate of ischemic heart disease was 327 cases/100,000 person-years, corresponding to 470 cases/100,000 for men and 211 cases/100,000 people-years for women. The classical cardiovascular risk factors that were associated with a higher risk of presenting events in the follow-up were dyslipidemia and arterial hypertension, in addition to male sex and age. Conclusions: The incidence of ischemic heart disease in the urban–rural population of Extremadura is high. The classical cardiovascular risk factors most strongly associated with events were dyslipidemia, and arterial hypertension (AU)

Myocardial infarction in older than 75 years: An increasing population. CASTUO Study. / Infarto de miocardio en mayores de 75 años: una población en aumento. Estudio CASTUO.

OBJECTIVES: Elderly patients with acute myocardial infarction constitute a population that is not adequately represented in clinical trials or medical registries. Our objective was to compare the clinical characteristics, treatments administered and mortality among patients younger and older than 75 years. MATERIAL AND METHODS: Observational retrospective study of patients hospitalized for acute myocardial infarction in the decade 2000-2009. Multivariate models were constructed to determine hospital and late mortality (median, 4.6 years; IQR 25-75: 2.1-7.3). RESULTS: We included 2,177 patients (995 men [79%]), with a mean age of 70.8 years (SD, 12.6). A total of 917 (42.0%) of the patients were 75 years of age or older. When compared with the patients younger than 75 years, the older patients had a greater prevalence of diabetes (38.3% vs. 32.5%; P<.002), chronic obstructive pulmonary disease (15.6% vs. 11.2%; P<.002), stroke (14.3% vs. 7.3%; P<.001), chronic renal failure (11.0% vs. 3.9%; P<.001), atrial fibrillation (15.9% vs. 6.9%; P<.001), heart failure (28.0% vs. 23.4%; P<.008). The older patients were treated with fewer beta-blockers (55.9% vs. 71.2%; P<.001), statins (44.3% vs. 62.3%; P<.001), coronary angiographies (17.9% vs. 48.5%; P<.001) and angioplasties (10.8% vs. 29.1%; P<.001). The patients older than 75 years had lower survival (mortality, 44.5% vs. 18.9%; HR 1.89; 95% CI 1.57-2.29). The use of beta-blockers (HR, 0.74; 95% CI 0.62-0.89), statins (HR 0.73; 95% CI 0.58-0.91) and angioplasty (HR, 0.42; 95% CI 0.30-0.57) was inversely correlated with mortality. CONCLUSIONS: Patients older than 75 years with acute myocardial infarction had lower survival and were treated with fewer beta-blockers, statins and angioplasty, indications that are associated with lower mortality.

Prevalence of atrial fibrillation and its associated factors in Spain: An analysis of 6 population-based studies. DARIOS Study. / Prevalencia de fibrilación auricular y factores asociados en España, análisis de seis estudios de base poblacional. Estudio DARIOS.

BACKGROUND AND OBJECTIVES: Atrial fibrillation (AF) is the most common type of arrhythmia. The purpose of this study was to determine the prevalence of atrial fibrillation and its relationship with cardiovascular risk factors in Spain. METHODOLOGY: Cross-sectional study based on a grouped analysis of 17,291 randomized individuals recruited in 6 population studies. RESULTS: The prevalence of atrial fibrillation was 1.5% (95% CI:1.3-1.7%). Men had a greater prevalence of the disease than women (1.9 vs. 1.1%, respectively). The prevalence of atrial fibrillation progressively increased with age: 0.05% for patients younger than 45 years, 0.5% for those between 45-59 years of age, 2.3% for those between 60-74 years of age and 6.3% for those older than 75 years. The percentage of individuals who were underwent anticoagulant treatment was 74.3%. The risk factors significantly associated with arrhythmia were an age older than 60 years (odds ratio [OR]: 7.6; 95% CI: 5.1-11.2), the male sex (OR:1.8; 95% CI: 1.4-2.4), arterial hypertension (OR:1.6; 95% CI: 1.2-2.1), obesity (OR:1.5; 95% CI:1.2-2.1) and a history of coronary artery disease (OR:1.9; 95% CI: 1.3-3.0). CONCLUSION: Atrial fibrillation is a common disease in elderly individuals, while its prevalence is low in individuals younger than 60 years. Most individuals with atrial fibrillation were on anticoagulant treatment. The risk factors for this type of arrhythmia are age, the male sex, hypertension, obesity and a history of coronary artery disease.

Ischemic stroke mortality tendency (2000-2009) and prognostic factors. ICTUS Study-Extremadura (Spain).

BACKGROUND AND OBJECTIVES: Mortality due to stroke is high in our setting. However, we do not know its magnitude and course in a recent decade. Thus, we have assessed the global inhospital mortality and that at 1 year of stroke in patients seen in a regional hospital as well as its prognostic factors. MATERIAL AND METHODS: A one-year follow-up historical cohort study was performed of patients admitted at Don Benito-Villanueva de la Serena Regional Hospital (Badajoz) with diagnosis of ischemic stroke (1 January 2000 to 31 December 2009). Epidemiological, clinical data all cause death information were collected. RESULTS: A total of 2.228 patients (50.8% male), mean age 71 (SD 10) years were recruited. In-hospital mortality rate was 15.3% and mortality rate at 1 year was 16.9%, with no significant changes during the 10-year study period. Risk factors for greater in-hospital mortality were age, previous renal and heart failure and not performing diagnostic tests. Factors associated with 1-year all-cause mortality were age, dementia, chronic obstructive pulmonary disease, myocardial infarction, atrial fibrillation and also not performing diagnostic tests. During the study, use of diagnostic tests and treatment with statin, heparin and antihypertensive medication on discharge increased. CONCLUSIONS: Patients admitted due to ischemic stroke, in a regional hospital presented a 1 year mortality rate of 29.6%. This tendency did not improve during the 10-year study period. Mortality was associated to greater age, comorbidities and not performing diagnostic tests.

Clinical characteristics and mortality of heart failure. INCAex study.

BACKGROUND AND OBJECTIVES: Heart failure is responsible for a major part of hospital health expenditure and the third cause of cardiovascular death. To describe the evolution of clinical features, and factors related to prognosis of patients admitted due to decompensated heart failure in a region of Extremadura during a period 10 years. PATIENTS AND METHODS: Observational, retrospective and single centre study of consecutive patients admitted due to decompensated heart failure in a general hospital in the province of Badajoz, during the period 2000/2009. RESULTS: A total of 2220 patients with mean age of 76.3 (SD±10.1), being 54% female were included in the study. Stratified into four periods (30 months each), a significant increase in patients over 75 years was observed (55 vs. 71%; P<.001), as well as an increase in the prescription of beta blockers at discharge (12 vs. 34%, P<.001), statins (8 vs. 31%; P<.001), and oral anticoagulants (13 vs. 25%; P<.001). Hospital mortality significantly decreased from 13 to 8% (P<.01), and from 30 to 23% (P<.01) at one year follow-up. Age (HR per year=1.04 [95% CI: 1.02 to 1.05]), diabetes (HR=1.35 [95% CI: 1.11 to 1.66]) and chronic renal failure (HR=1.49 [95% CI: 1.18 to 1.87]) were identified as independent predictors of all-cause mortality at one year of follow-up. CONCLUSIONS: Total mortality in patients with decompensated heart failure has declined significantly over the last decade, despite the increasing age. Age, diabetes and chronic renal failure were independent predictors of total mortality at one year. Oral anticoagulation was a protective factor.

Transgenic Medicago truncatula plants that accumulate proline display nitrogen-fixing activity with enhanced tolerance to osmotic stress.

Legume root nodule nitrogen-fixing activity is severely affected by osmotic stress. Proline accumulation has been shown to induce tolerance to salt stress, and transgenic plants over-expressing Delta(1)-pyrroline-5-carboxylate synthetase (P5CS), which accumulates high levels of proline, display enhanced osmotolerance. Here, we transformed the model legume Medicago truncatula with the P5CS gene from Vigna aconitifolia, and nodule activity was evaluated under osmotic stress in transgenic plants that showed high proline accumulation levels. Nitrogen fixation was significantly less affected by salt treatment compared to wild-type (WT) plants. To our knowledge, this is the first time that transgenic legumes have been produced that display nitrogen-fixing activity with enhanced tolerance to osmotic stress. We studied the expression of M. truncatula proline-related endogenous genes M. truncatulaDelta(1)-pyrroline-5-carboxylate synthetase 1 (MtP5CS1), M. truncatulaDelta(1)-pyrroline-5-carboxylate synthetase 2 (MtP5CS2), M. truncatula ornithine delta-aminotransferase (MtOAT), M. truncatula proline dehydrogenase (MtProDH) and a proline transporter gene in both WT and transgenic plants. Our results indicate that proline metabolism is finely regulated in response to osmotic stress in an organ-specific manner. The transgenic model allowed us to analyse some of the biochemical and molecular mechanisms that are activated in the nodule in response to high salt conditions, and to ascertain the essential role of proline in the maintenance of nitrogen-fixing activity under osmotic stress.

Biotransformation of tryptamine and secologanin into plant terpenoid indole alkaloids by transgenic yeast.

A transgenic Saccharomyces cerevisiae was constructed containing the cDNAs coding for strictosidine synthase (STR) and strictosidine beta-glucosidase (SGD) from the medicinal plant Catharanthus roseus. Both enzymes are involved in the biosynthesis of terpenoid indole alkaloids. The yeast culture was found to express high levels of both enzymes. STR activity was found both inside the cells (13.2 nkatal/g fresh weight) and in the medium (up to 25 nkatal/l medium), whereas SGD activity was present only inside the yeast cells (2.5 mkatal/g fresh weight). Upon feeding of tryptamine and secologanin, this transgenic yeast culture produced high levels of strictosidine in the medium; levels up to 2 g/l were measured. Inside the yeast cells strictosidine was also detected, although in much lower amounts (0.2 mg/g cells). This was due to the low permeability of the cells towards the substrates, secologanin and tryptamine. However, the strictosidine present in the medium was completely hydrolyzed to cathenamine, after permeabilizing the yeast cells. Furthermore, transgenic S. cerevisiae was able to grow on an extract of Symphoricarpus albus berries serving as a source for secologanin and carbohydrates. Under these conditions, the addition of tryptamine was sufficient for the transgenic yeast culture to produce indole alkaloids. Our results show that transgenic yeast cultures are an interesting alternative for the production of plant alkaloids.