The fast-growing Brucella suis Biovar 5 depends on phosphoenolpyruvate carboxykinase and pyruvate phosphate dikinase but not on Fbp and GlpX fructose-1, 6-bisphosphatases or isocitrate lyase for full virulence in laboratory models

Bacteria of the genus Brucella infect a range of vertebrates causing a worldwide extended zoonosis. The best-characterized brucellae infect domestic livestock, behaving as stealthy facultative intracellular parasites. This stealthiness depends on envelope molecules with reduced pathogen-associated molecular patterns, as revealed by the low lethality and ability to persist in mice of these bacteria. Infected cells are often engorged with brucellae without signs of distress, suggesting that stealthiness could also reflect an adaptation of the parasite metabolism to use local nutrients without harming the cell. To investigate this, we compared key metabolic abilities of Brucella abortus 2308 Wisconsin (2308W), a cattle biovar 1 virulent strain, and B. suis 513, the reference strain of the ancestral biovar 5 found in wild rodents. B. suis 513 used a larger number of C substrates and showed faster growth rates in vitro, two features similar to those of B. microti, a species phylogenomically close to B. suis biovar 5 that infects voles. However, whereas B. microti shows enhanced lethality and reduced persistence in mice, B. suis 513 was similar to B. abortus 2308W in this regard. Mutant analyses showed that B. suis 513 and B. abortus 2308W were similar in that both depend on phosphoenolpyruvate synthesis for virulence but not on the classical gluconeogenic fructose-1, 6-bisphosphatases Fbp-GlpX or on isocitrate lyase (AceA). However, B. suis 513 used pyruvate phosphate dikinase (PpdK) and phosphoenolpyruvate carboxykinase (PckA) for phosphoenolpyruvate synthesis in vitro while B. abortus 2308W used only PpdK. Moreover, whereas PpdK dysfunction causes attenuation of B. abortus 2308W in mice, in B. suis, 513 attenuation occurred only in the double PckA-PpdK mutant. Also contrary to what occurs in B. abortus 2308, a B. suis 513 malic enzyme (Mae) mutant was not attenuated, and this independence of Mae and the role of PpdK was confirmed by the lack of attenuation of a double Mae-PckA mutant. Altogether, these results decouple fast growth rates from enhanced mouse lethality in the brucellae and suggest that an Fbp-GlpX-independent gluconeogenic mechanism is ancestral in this group and show differences in central C metabolic steps that may reflect a progressive adaptation to intracellular growth

Influence of Phragmites australis on the seasonal evolution of biogeochemical conditions (pH/Eh) and N cycling in eutrophic wetlands

[SPA] El objetivo del trabajo fue evaluar en qué medida humedales eutrofizados, en presencia y ausencia de Phragmites australis, actúan como depuradores de NO3- y productores de N2O (gas de efecto invernadero). Para ello se realizó un experimento en mesocosmos con suelo y plantas de Phragmites, que se sometieron a seis períodos de inundación-desecación durante doce meses con aguas de distinta composición: alta carga (200 mg·L-1 NO3-) y baja carga (20 mg·L-1 NO3-) de nutrientes. Los tratamientos ensayados fueron: sin planta+baja carga; sin planta+alta carga; con planta+baja carga; con planta+alta carga. Se monitorizó Eh y pH, y se analizó la concentración de NO3- y NH4+ en la solución edáfica así como el N2O desprendido a la atmósfera. Todos los tratamientos fueron efectivos en la retirada de NO3- del agua (>70%) lo que acarreó la emisión de elevadas cantidades de N2O a la atmósfera. Phragmites contribuyó a reducir las emisiones de N2O durante las fases de secado al favorecer la retirada de NO3- del agua y disminuir su disponibilidad para ser transformado en gas vía desnitrificación. Por tanto, la presencia de planta en los suelos de los humedales puede contribuir a que se reduzca la emisión de N2O a la atmósfera. [ENG] The aim of the study was to assess to what extend eutrophic wetlands, with and without Phragmites australis, act as sinks of NO3- and as sources of N2O (greenhouse gas). A one-year mesocosms experiment, with alternating flooding-drying conditions, was performed with soil and plants of Phragmites. Two types of water were used: one with high nutrient load (200 mg·L-1 NO3-) and another one with low nutrient load (20 mg·L-1 NO3-). The treatments assayed were: no plant+low nutrient content; no plant+high nutrient content; plant+low nutrient content; plant+high nutrient content. The Eh and pH were monitored, and the concentration of NO3- and NH4+ in the soil solution and of evolved N2O were analyzed. All the treatments were effective in the removal of NO3- from water (>70%), leading to the emission of high amounts of N2O into the atmosphere. The presence of Phragmites favored lower N2O emissions mainly during drying phases due to a higher reduction of NO3- concentrations in water, decreasing its availability to be transformed into gas via denitrification. Therefore, the presence of plant in wetland soils may contribute to reduce the emission of N2O into the atmosphere.Proyecto financiado por el Ministerio de Ciencia e Innovación (Referencia CGL2010-20214)

Molecular profiling of immunoglobulin heavy-chain gene rearrangements unveils new potential prognostic markers for multiple myeloma patients

Altres ajuts: This work was partially supported by[...] , CIBERONC-CB16/12/00233, and "Una manera de hacer Europa" (Innocampus; CEI-2010-1-0010)". M.G.-A., I.P.-C., and C.J. are supported by the Fundación Española de Hematología y Hemoterapia (FEHH, co-funded by Fundación Cris in the latter case), A.M. by the European Social Fund and the Spanish Education Council through the University of Salamanca, [...]. All Spanish funding is co-sponsored by the European Union FEDER program.Multiple myeloma is a heterogeneous disease whose pathogenesis has not been completely elucidated. Although B-cell receptors play a crucial role in myeloma pathogenesis, the impact of clonal immunoglobulin heavy-chain features in the outcome has not been extensively explored. Here we present the characterization of complete heavy-chain gene rearrangements in 413 myeloma patients treated in Spanish trials, including 113 patients characterized by next-generation sequencing. Compared to the normal B-cell repertoire, gene selection was biased in myeloma, with significant overrepresentation of IGHV3, IGHD2 and IGHD3, as well as IGHJ4 gene groups. Hypermutation was high in our patients (median: 8.8%). Interestingly, regarding patients who are not candidates for transplantation, a high hypermutation rate (≥7%) and the use of IGHD2 and IGHD3 groups were associated with improved prognostic features and longer survival rates in the univariate analyses. Multivariate analysis revealed prolonged progression-free survival rates for patients using IGHD2/IGHD3 groups (HR: 0.552, 95% CI: 0.361−0.845, p = 0.006), as well as prolonged overall survival rates for patients with hypermutation ≥7% (HR: 0.291, 95% CI: 0.137−0.618, p = 0.001). Our results provide new insights into the molecular characterization of multiple myeloma, highlighting the need to evaluate some of these clonal rearrangement characteristics as new potential prognostic markers

Biotecnología de plantas superiores y algas verdes

Los ureidos, alantoína y alantoato, son los principales compuestos sintetizados a partir del nitrógeno fijado en los nódulos de las leguminosas que se exportan a las partes aéreas en leguminosas tropicales como la judía. Los ureidos se producen por la oxidación de las purinas sintetizadas de novo en los nódulos radicales y también como parte de un proceso de recuperación de los compuestos nitrogenados en tejidos senescentes. Los ureidos se acumulan en varios tejidos vegetales en respuesta al estrés hídrico, y se ha sugerido que la acumulación de estos compuestos nitrogenados es la responsable de la inhibición de la fijación de nitrógeno que tiene lugar en condiciones ambientales adversas. A pesar de la importancia crucial de los ureidos como compuestos de reserva y transporte de nitrógeno, hasta el momento no se llevado a cabo la caracterización de las rutas de síntesis y degradación de ureidos en plantas. En particular, parece que existen dos posibles rutas de degradación de alantoato, y que el que determinadas leguminosas usen una u otra ruta puede afectar a la sensibilidad o tolerancia de esas plantas a la sequía. Por tanto la determinación de la ruta que opera en leguminosas de gran interés agronómico como la judía y el garbanzo, puede a la larga tener utilidad biotecnológica con la obtención de plantas que sean capaces de mantener la fijación de nitrógeno en condiciones ambientales adversas, como la sequía. Además de en la caracterización, con fines biotecnológicos del metabolismo de ureidos en leguminosas, se está trabajando obtención de algas superproductoras de γ-tocoferol (vitamina E) mediante ingeniería metabólica, así como en el análisis del contenido en antioxidantes del aceite de oliv

Modifiable risk factors associated with prediabetes in men and women: A cross-sectional analysis of the cohort study in primary health care on the evolution of patients with prediabetes

Background: Prediabetes is a high-risk state for diabetes development, but little is known about the factors associated with this state. The aim of the study was to identify modifiable risk factors associated with the presence of prediabetes in men and women. Methods: Cohort Study in Primary Health Care on the Evolution of Patients with Prediabetes (PREDAPS-Study) is a prospective study on a cohort of 1184 subjects with prediabetes and another cohort of 838 subjects without glucose metabolism disorders. It is being conducted by 125 general practitioners in Spain. Data for this analysis were collected during the baseline stage in 2012. The modifiable risk factors included were: smoking habit, alcohol consumption, low physical activity, inadequate diet, hypertension, dyslipidemia, and obesity. To assess independent association between each factor and prediabetes, odds ratios (ORs) were estimated using logistic regression models. Results: Abdominal obesity, low plasma levels of high-density lipoprotein cholesterol (HDL-cholesterol), and hypertension were independently associated with the presence of prediabetes in both men and women. After adjusting for all factors, the respective ORs (95% Confidence Intervals) were 1.98 (1.41-2.79), 1.88 (1.23-2.88) and 1.86 (1.39-2.51) for men, and 1.89 (1.36-2.62), 1.58 (1.12-2.23) and 1.44 (1.07-1.92) for women. Also, general obesity was a risk factor in both sexes but did not reach statistical significance among men, after adjusting for all factors. Risky alcohol consumption was a risk factor for prediabetes in men, OR 1.49 (1.00-2.24). Conclusions: Obesity, low HDL-cholesterol levels, and hypertension were modifiable risk factors independently related to the presence of prediabetes in both sexes. The magnitudes of the associations were stronger for men than women. Abdominal obesity in both men and women displayed the strongest association with prediabetes. The findings suggest that there are some differences between men and women, which should be taken into account when implementing specific recommendations to prevent or delay the onset of diabetes in adult population

Seed storage conditions change the germination pattern of clonal growth plants in Mediterranean salt marshes.

8 páginas, 4 tablas, 8 figuras.The effect of salinity level and extended exposure to different salinity and flooding conditions on germination patterns of three saltmarsh clonal growth plants (Juncus subulatus, Scirpus litoralis, and S. maritimus) was studied. Seed exposure to extended flooding and saline conditions significantly affected the outcome of the germination process in a different, though predictable, way for each species, after favorable conditions for germination were restored. Tolerance of the germination process was related to the average salinity level measured during the growth/germination season at sites where established individuals of each species dominated the species cover. No relationship was found between salinity tolerance of the germination process and seed response to extended exposure to flooding and salinity conditions. The salinity response was significantly related to the conditions prevailing in the habitats of the respective species during the unfavorable (nongrowth/nongermination) season. Our results indicate that changes in salinity and hydrology while seeds are dormant affect the outcome of the seed-bank response, even when conditions at germination are identical. Because these environmental-history-dependent responses differentially affect seed germination, seedling density, and probably sexual recruitment in the studied and related species, these influences should be considered for wetland restoration and managementFinancial support from the Spanish Ministry of the Environment (MMA, project 05/99) and the Junta de Andalucía (research group 4086)enabled us to carry out the present work.Peer reviewe

Seed storage conditions change the germination pattern of clonal growth plants in Mediterranean salt marshes.

8 páginas, 4 tablas, 8 figuras.The effect of salinity level and extended exposure to different salinity and flooding conditions on germination patterns of three saltmarsh clonal growth plants (Juncus subulatus, Scirpus litoralis, and S. maritimus) was studied. Seed exposure to extended flooding and saline conditions significantly affected the outcome of the germination process in a different, though predictable, way for each species, after favorable conditions for germination were restored. Tolerance of the germination process was related to the average salinity level measured during the growth/germination season at sites where established individuals of each species dominated the species cover. No relationship was found between salinity tolerance of the germination process and seed response to extended exposure to flooding and salinity conditions. The salinity response was significantly related to the conditions prevailing in the habitats of the respective species during the unfavorable (nongrowth/nongermination) season. Our results indicate that changes in salinity and hydrology while seeds are dormant affect the outcome of the seed-bank response, even when conditions at germination are identical. Because these environmental-history-dependent responses differentially affect seed germination, seedling density, and probably sexual recruitment in the studied and related species, these influences should be considered for wetland restoration and managementFinancial support from the Spanish Ministry of the Environment (MMA, project 05/99) and the Junta de Andalucía (research group 4086)enabled us to carry out the present work.Peer reviewe

La secuenciación masiva dirigida revela que los pacientes con leucemia linfática crónica y reordenamiento de igh presentan mutaciones en los genes POT1, EGR2, BRAF, IGLL5 Y MGA

Abstract [CO-081] Introducción: La traslocación de la región 14q32, que contiene el gen de la cadena pesada de las inmunoglobulinas (IGH), aparece en el 4-9% de pacientes de leucemia linfática crónica(LLC). Aunque algunos estudios le atribuyen a este subgrupo un pronóstico desfavorable, sus características clínicas y biológicas no se conocen en profundidad. La secuenciación masiva (NGS) ha mejorado notablemente el conocimiento de la heterogeneidad genética y clínica de la LLC, por lo que nos planteamos el análisis del perfil mutacional de estos pacientes para definir mejor su pronóstico. Métodos: Se analizaron 231 pacientes de LLC, de los cuales 42 presentaban traslocación de 14q32. En todos los casos se disponía de datos clínicos y FISH. Se diseñó un panel personalizado de 54 genes, seleccionados por su frecuencia e implicación en la patogenia de la enfermedad. La secuenciación se realizó en la plataforma NextSeq(Illumina). El panel cubre el 97% de las regiones (>100X) con una profundidad de 606 lecturas/base, permitiendo la detección de variantes presentes en >3% de las células..

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