Inversion of the anomeric configuration of the transferred sugar during inactivation of the macrolide antibiotic oleandomycin catalyzed by a macrolide glycosyltransferase

AbstractMacrolides are a group of antibiotics structurally characterized by a macrocyclic lactone to which one or several deoxy-sugar moieties are attached. The sugar moieties are transferred to the different aglycones by glycosyltransferases (GTF). The OleI GTF of an oleandomycin producer, Streptomyces antibioticus, catalyzes the inactivation of this macrolide by glycosylation. The product of this reaction was isolated and its structure elucidated. The donor substrate of the reaction was UDP-α-D-glucose, but the reaction product showed a β-glycosidic linkage. The inversion of the anomeric configuration of the transferred sugar and other data about the kinetics of the reaction and primary structure analysis of several GTFs are compatible with a reaction mechanism involving a single nucleophilic substitution at the sugar anomeric carbon in the catalytic center of the enzyme

Robotized spraying of prefabricated panels

A robotic manufacturing cell of pre-fabricated glass reinforced cement panels for construction industry has been developed by DISAM for the Spanish construction company Dragados, SA. The main contribution of the developed system is the automatic programming and control of the whole plan. The architect's 3D-drawing of the building facade done on a CAD system serves as input. From the CAD design, the optimum facade to panels partition is obtained. In order to manufacture each panel, automatic task and path planning are performed for the equipment present in the manufacturing cell: spraying robot, PLCs, control computer, etcThis work was supported by the construction company Dragados, S.A. and the Spanish Ministry of Industry under project PAUTA 1691/91. The authors thank A. Garcia, E. Pinto, 1. Florez, E. Marquez, C. Corpas, 1. Arauzo and A. Cases, and the staff of the Caracola factory in Torrejon de Ardoz (Madrid). Thanks are also due to Christian Schafer for his assistance with the final manuscript.Publicad

Optimization of the Bioactivation of Isoflavones in Soymilk by Lactic Acid Bacteria

Soybeans and soy-based products contain isoflavones which can be used for nutraceutical and medical applications. In soybeans and in unfermented soy foods, isoflavones are normally present as glycosides. Isoflavone glycosides can be enzymatically converted to isoflavone aglycones, thus releasing the sugar molecule. The effective absorption of isoflavones in humans requires the bioconversion of isoflavone glycosides to isoflavone aglycones through the activity of the enzyme β-glucosidase. The objective was to assess the capacity of 42 bacterial strains (belonging to Lactobacillus, Streptococcus and Enterococcus) to produce β-glucosidase activity. The strain that showed the highest β-glucosidase activity (Lactobacillus plantarum 128/2) was then used for the optimization of the bioconversion of genistin and daidzin present in commercial soymilk to their aglycone forms genistein and daidzein. The contribution of process parameters (temperature, inoculum size, time) to the efficiency of such bioactivation was tested. Lactobacillus plantarum 128/2 was able to completely bioactivate soymilk isoflavones under the following conditions: 25 °C temperature, 2% inoculum size and 48 h process time. These results confirm the suitability of lactic acid bacteria for the bioactivation of isoflavones present in soymilk and provide an interesting candidate (L. plantarum 182/2) for food industries to perform this transformation.Jon Kepa Izaguirre was the recipient of a predoctoral fellowship from the Basque Government

Cardiotrophin-1 opposes renal fibrosis in mice: Potential prevention of chronic kidney disease

[EN]Chronic kidney disease is characterized by tubulointerstitial fibrosis involving inflammation, tubular apoptosis, fibroblast proliferation and extracellular matrix accumulation. Cardiotrophin-1, a member of the interleukin-6 family of cytokines, protects several organs from damage by promoting survival and anti-inflammatory effects. However, whether cardiotrophin-1 participates in the response to chronic kidney injury leading to renal fibrosis is unknown. We hypothesized and assessed the potential role of cardiotrophin-1 in a mice model of tubulointerstitial fibrosis induced by unilateral ureteral obstruction (UUO). Three days after UUO, obstructed kidneys from cardiotrophin-1-/- mice show higher expression of inflammatory markers IL-1β, Cd68, ICAM-1, COX-2 and iNOs, higher activation of NF-κB, higher amount of myofibroblasts and higher severity of tubular damage and apoptosis, compared with obstructed kidneys from wild-type littermates. In a later stage, obstructed kidneys from cardiotrophin-1-/- mice show higher fibrosis than obstructed kidneys from wild-type mice. Interestingly, administration of exogenous cardiotrophin-1 prevents the increased fibrosis resulting from the genetic knockout of cardiotrophin-1 upon UUO, and supplementation of wild-type mice with exogenous cardiotrophin-1 further reduces the renal fibrosis induced by UUO. In vitro, renal myofibroblasts from cardiotrophin-1-/- mice have higher collagen I and fibronectin expression and higher NF-κB activation than wild-type cells. Cardiotrophin-1 participates in the endogenous response that opposes renal damage by counteracting the inflammatory, apoptotic and fibrotic processes. And exogenous cardiotrophin-1 is proposed as a candidate for the treatment and prevention of chronic renal fibrosis

Metabolite profile of Nectandra oppositifolia Nees & Mart. and assessment of antitrypanosomal activity of bioactive compounds through efficiency analyses

EtOH extracts from the leaves and twigs of Nectandra oppositifolia Nees & Mart. shown activity against amastigote forms of Trypanosoma cruzi. These extracts were subjected to successive liquid-liquid partitioning to afford bioactive CH2Cl2 fractions. UHPLC-TOF-HRMS/MS and molecular networking were used to obtain an overview of the phytochemical composition of these active fractions. Aiming to isolate the active compounds, both CH2Cl2 fractions were subjected to fractionation using medium pressure chromatography combined with semi-preparative HPLC-UV. Using this approach, twelve compounds (1-12) were isolated and identified by NMR and HRMS analysis. Several isolated compounds displayed activity against the amastigote forms of T. cruzi, especially ethyl protocatechuate (7) with EC50 value of 18.1 μM, similar to positive control benznidazole (18.7 μM). Considering the potential of compound 7, protocatechuic acid and its respective methyl (7a), n-propyl (7b), n-butyl (7c), n-pentyl (7d), and n-hexyl (7e) esters were tested. Regarding antitrypanosomal activity, protocatechuic acid and compound 7a were inactive, while 7b-7e exhibited EC50 values from 20.4 to 11.7 μM, without cytotoxicity to mammalian cells. These results suggest that lipophilicity and molecular complexity play an important role in the activity while efficiency analysis indicates that the natural compound 7 is a promising prototype for further modifications to obtain compounds effective against the intracellular forms of T. cruzi

Prevalence of hyponatraemia in patients over the age of 65 who have an in-hospital fall

Background and aim: Hyponatraemia is the most common electrolyte disorder. Some studies have found that it increases morbidity and mortality. There are new lines of research that are investigating the link between hyponatraemia and patient falls. Aim: To determine if hyponatraemia is associated with falls in elderly hospitalised patients. Methods: Design observational, analytical, case-control study. Study population: Patients older than 65 years who had fallen during their hospitalisation at Gregorio Marañón Hospital (Madrid) were considered cases. Patients who did not fall were considered to be controls, paired according to the following variables: hospital ward, age, length of hospital stay, gender and Downton fall risk index. The sample size was 206 subjects. Data collection: Socio-demographic factors, variables included in the falls record sheet, Downton fall risk index and sodium levels were studied (hyponatraemia was considered Na+&lt; 135 mmol/l). Analysis: A descriptive analysis was performed to determine the sample homogeneity. The OR was calculated, and an analytical analysis using Chi-square test and a multivariate logistic regression analysis were also performed. Results: Of 103 cases recruited, 61 were men (50.4%) and 42 were women (49.4%). Hyponatraemia was detected in 29 cases with an association with falls of P: 0.002. The adjusted OR was 3.708 (1.6-8.3), 95% CI. Risk factors for falls were identified as hyponatraemia and limb sensory deficits. Conclusions: Given that hyponatraemia could be considered a risk factor for falls, the inclusion of the determination of sodium level would be important for fall prevention strategies in the elderly.Fundamento y objetivo: La hiponatremia es el trastorno electrolítico más frecuente. Algunos estudios afirman que aumenta la morbimortalidad. Existen nuevas líneas de investigación que buscan la relación entre hiponatremia y caídas. Objetivo: Determinar si la hiponatremia es un factor relacionado con las caídas en ancianos hospitalizados. Método: Disen˜ o observacional analítico de casos y controles. Población de estudio: Se consideraron casos los pacientes mayores de 65 an˜ os que experimentaron una caída durante su ingreso en unidades de hospitalización del Hospital General Universitario Gregorio Maran˜ ón de Madrid. Los controles fueron pacientes que no wxperimentaron caída, pareados según las variables: unidad, edad, periodo de ingreso, género y Downton. El taman˜ o fue de 206 sujetos. Recogida de datos: Se estudiaron factores sociodemográficos, las variables incluidas en la ficha de registro de caídas y escala de Downton, y el sodio sérico. Se consideró hiponatremia Na+ < 135 mmol/l. Análisis: Se realizó un análisis descriptivo para valorar la homogeneidad de la muestra, un análisis analítico utilizando el test chi cuadrado, calculando la OR y un análisis multivariante con regresión logística. Resultados: De 103 casos, 61 eran hombres (50,4%) y 42 mujeres (49,4%). En 29 se detectó hiponatremia; la relación con las caídas fue p: 0,002. La OR ajustada fue de 3,708 (1,6-8,3), IC 95%. Se identificaron como factores de riesgo para las caídas: hiponatremia y déficits sensoriales en extremidade

Marine megafauna niche coexistence and hotspot areas in a temperate ecosystem

Versión del editor2,08

Safety and vaccine-induced HIV-1 immune responses in healthy volunteers following a late MVA-B boost 4 years after the last immunization

Background: We have previously shown that an HIV vaccine regimen including three doses of HIV-modified vaccinia virus Ankara vector expressing HIV-1 antigens from clade B (MVA-B) was safe and elicited moderate and durable (1 year) T-cell and antibody responses in 75% and 95% of HIV-negative volunteers (n = 24), respectively (RISVAC02 study). Here, we describe the long-term durability of vaccine-induced responses and the safety and immunogenicity of an additional MVA-B boost. Methods: 13 volunteers from the RISVAC02 trial were recruited to receive a fourth dose of MVA-B 4 years after the last immunization. End-points were safety, cellular and humoral immune responses to HIV-1 and vector antigens assessed by ELISPOT, intracellular cytokine staining (ICS) and ELISA performed before and 2, 4 and 12 weeks after receiving the boost. Results: Volunteers reported 64 adverse events (AEs), although none was a vaccine-related serious AE. After 4 years from the 1st dose of the vaccine, only 2 volunteers maintained low HIV-specific T-cell responses. After the late MVA-B boost, a modest increase in IFN-γ T-cell responses, mainly directed against Env, was detected by ELISPOT in 5/13 (38%) volunteers. ICS confirmed similar results with 45% of volunteers showing that CD4+ T-cell responses were mainly directed against Env, whereas CD8+ T cell-responses were similarly distributed against Env, Gag and GPN. In terms of antibody responses, 23.1% of the vaccinees had detectable Env-specific binding antibodies 4 years after the last MVA-B immunization with a mean titer of 96.5. The late MVA-B boost significantly improved both the response rate (92.3%) and the magnitude of the systemic binding antibodies to gp120 (mean titer of 11460). HIV-1 neutralizing antibodies were also enhanced and detected in 77% of volunteers. Moreover, MVA vector-specific T cell and antibody responses were boosted in 80% and 100% of volunteers respectively. Conclusions: One boost of MVA-B four years after receiving 3 doses of the same vaccine was safe, induced moderate increases in HIV-specific T cell responses in 38% of volunteers but significantly boosted the binding and neutralizing antibody responses to HIV-1 and to the MVA vector