Enhanced polyhydroxyalkanoates accumulation by Halomonas spp. in artificial biofilms of alginate beads.

Microbial mats are complex but stable, multi-layered and multi-functional biofilms, which are the most frequent bacterial formations in nature. The functional strategies and physiological versatility of the bacterial populations growing in microbial mats allow bacteria to resist changing conditions within their environment. One of these strategies is the accumulation of carbon- and energy-rich polymers that permit the recovery of metabolic activities when favorable conditions are restored. In the present study, we systematically screened microbial mats for bacteria able to accumulate large amounts of the ester carbon polymers polyhydroxyalkanoates (PHA). Several of these strains were isolated from Ebro Delta microbial mats and their ability to accumulate PHA up to 40-60 % of their dry weight was confirmed. According to two identification approaches (16S rRNA and ropD genes), these strains were identified as Halomonas alkaliphila (MAT-7, -13, -16), H. neptunia (MAT-17), and H. venusta (MAT-28). To determine the mode of growth yielding maximum PHA accumulation, these three different species were cultured in an artificial biofilm in which the cells were immobilized on alginate beads. PHA accumulation by cells that had detached from the biofilm was compared with that of their planktonic counterparts. Experiments in different culture media showed that PHA accumulation, measured as the relative fluorescence intensity after 48 h of incubation at 30 °C, was higher in immobilized than in planktonic cells, with the exception of cells growing in 5 % NaCl, in which PHA accumulation was drastically lower in both. Therefore, for obtaining high PHA concentrations, the use of immobilized cells may be a good alternative to the PHA accumulation by bacteria growing in the classical, planktonic mode. From the ecological point of view, increased PHA accumulation in detached cells from biofilms would be a natural strategy to improve bacterial dispersion capacity and, consequently, to increase survival in stressed environments.

Mecánica de la succión: comparación entre la lactancia artificial y la lactancia materna

Introducción: Hay poco conocimiento sobre la similitud de la mecánica entre la lactancia materna y la artificial. Evaluamos la mecánica de la succión en neonatos con lactancia materna exclusiva, lactancia artificial exclusiva y lactancia mixta. Nuestra hipótesis fue que el patrón fisiológico de los movimientos de succión varía según el tipo de alimentación. Según esta hipótesis, los niños con lactancia materna exclusiva realizan unos movimientos al mamar distintos a los de la succión de una tetina, realizados por niños con lactancia artificial. Los niños con lactancia mixta mezclan ambos tipos de movimientos de succión. Métodos: Estudio transversal de neonatos de 21-28 días de edad con lactancia materna o artificial exclusiva (124 parejas madre-hijo), y ensayo de campo, abierto, cruzado y aleatorizado, realizado en neonatos de 21-28 días (110 parejas madre-hijo) y en lactantes de 3-5 meses de edad (125 parejas madre-hijo) con lactancia mixta. Las variables principales fueron los movimientos de succión y las pausas. Resultados: Los neonatos de 21-28 días de edad alimentados con lactancia artificial exclusiva mostraron un menor número de movimientos de succión y el mismo número de pausas, pero de mayor duración, que los neonatos con lactancia materna exclusiva. Entre los niños que recibieron lactancia mixta, el número de movimientos de succión al recibir alimentación con biberón fue similar y las pausas menos numerosas y de menor duración respecto a lo observado al amamantar, tanto a los 21-28 días como a los 3-5 meses de edad. En este grupo de lactancia mixta, la cifra media de tomas de lactancia materna fue de 5,83 ± 1,93 a los 21-28 días de vida y de 4,42 ± 1,67 a los 3-5 meses de edad. En el análisis de equivalencia, realizado sobre los niños que recibieron lactancia mixta, el intervalo de confianza del 95% de la razón de movimientos con lactancia artificial y con lactancia materna se situó fuera del rango de equivalencia, indicó un número de movimientos de succión menor en un 5,9-8,7% al tomar el biberón, así como un menor número de pausas y una duración más breve de ellas en este mismo grupo. Conclusiones: En la lactancia mixta, la comparación entre las tomas de biberón y las de pecho se situó fuera del rango de equivalencia, aunque las diferencias fueron pequeñas. Los niños con lactancia mixta mezclan ambos tipos de movimientos (lactancia materna y lactancia artificial) durante la fase de aprendizaje y adoptan su propio patrón.

Synthesis of triheptanoin and formulation as a solid diet for rodents

Triheptanoin-enriched diets have been successfully used in the experimental treatment of various metabolic disorders. Maximal therapeutic effect is achieved in the context of a ketogenic diet where triheptanoin oil provides 3040% of the daily caloric intake. However, pre-clinical studies using triheptanoin-rich diets are hindered by the difficulty of administering to laboratory animals as a solid foodstuff. In the present study, we successfully synthesized triheptanoin to the highest standards of purity from glycerol and heptanoic acid, using sulfonated charcoal as a catalyst. Triheptanoin oil was then formulated as a solid, stable and palatable preparation using a ketogenic base and a combination of four commercially available formulation agents: hydrophilic fumed silica, hydrophobic fumed silica, microcrystalline cellulose, and talc. Diet compliance and safety was tested on C57Bl/6 mice over a 15-week period, comparing overall status and body weight change. Practical applications: This work provides a complete description of (i) an efficient and cost-effective synthesis of triheptanoin and (ii) its formulation as a solid, stable, and palatable ketogenic diet (triheptanoin-rich; 39% of the caloric intake) for rodents. Triheptanoin-rich diets will be helpful on pre-clinical experiments testing the therapeutic efficacy of triheptanoin in different rodent models of human diseases. In addition, using the same solidification procedure, other oils could be incorporated into rodent ketogenic diet to study their dosage and long-term effects on mammal health and development. This approach could be extremely valuable as ketogenic diet is widely used clinically for epilepsy treatment.

Mecánica de la succión: comparación entre la lactancia artificial y la lactancia materna

Introducción: Hay poco conocimiento sobre la similitud de la mecánica entre la lactancia materna y la artificial. Evaluamos la mecánica de la succión en neonatos con lactancia materna exclusiva, lactancia artificial exclusiva y lactancia mixta. Nuestra hipótesis fue que el patrón fisiológico de los movimientos de succión varía según el tipo de alimentación. Según esta hipótesis, los niños con lactancia materna exclusiva realizan unos movimientos al mamar distintos a los de la succión de una tetina, realizados por niños con lactancia artificial. Los niños con lactancia mixta mezclan ambos tipos de movimientos de succión. Métodos: Estudio transversal de neonatos de 21-28 días de edad con lactancia materna o artificial exclusiva (124 parejas madre-hijo), y ensayo de campo, abierto, cruzado y aleatorizado, realizado en neonatos de 21-28 días (110 parejas madre-hijo) y en lactantes de 3-5 meses de edad (125 parejas madre-hijo) con lactancia mixta. Las variables principales fueron los movimientos de succión y las pausas. Resultados: Los neonatos de 21-28 días de edad alimentados con lactancia artificial exclusiva mostraron un menor número de movimientos de succión y el mismo número de pausas, pero de mayor duración, que los neonatos con lactancia materna exclusiva. Entre los niños que recibieron lactancia mixta, el número de movimientos de succión al recibir alimentación con biberón fue similar y las pausas menos numerosas y de menor duración respecto a lo observado al amamantar, tanto a los 21-28 días como a los 3-5 meses de edad. En este grupo de lactancia mixta, la cifra media de tomas de lactancia materna fue de 5,83 ± 1,93 a los 21-28 días de vida y de 4,42 ± 1,67 a los 3-5 meses de edad. En el análisis de equivalencia, realizado sobre los niños que recibieron lactancia mixta, el intervalo de confianza del 95% de la razón de movimientos con lactancia artificial y con lactancia materna se situó fuera del rango de equivalencia, indicó un número de movimientos de succión menor en un 5,9-8,7% al tomar el biberón, así como un menor número de pausas y una duración más breve de ellas en este mismo grupo. Conclusiones: En la lactancia mixta, la comparación entre las tomas de biberón y las de pecho se situó fuera del rango de equivalencia, aunque las diferencias fueron pequeñas. Los niños con lactancia mixta mezclan ambos tipos de movimientos (lactancia materna y lactancia artificial) durante la fase de aprendizaje y adoptan su propio patrón.

Synthesis of triheptanoin and formulation as a solid diet for rodents

Triheptanoin-enriched diets have been successfully used in the experimental treatment of various metabolic disorders. Maximal therapeutic effect is achieved in the context of a ketogenic diet where triheptanoin oil provides 3040% of the daily caloric intake. However, pre-clinical studies using triheptanoin-rich diets are hindered by the difficulty of administering to laboratory animals as a solid foodstuff. In the present study, we successfully synthesized triheptanoin to the highest standards of purity from glycerol and heptanoic acid, using sulfonated charcoal as a catalyst. Triheptanoin oil was then formulated as a solid, stable and palatable preparation using a ketogenic base and a combination of four commercially available formulation agents: hydrophilic fumed silica, hydrophobic fumed silica, microcrystalline cellulose, and talc. Diet compliance and safety was tested on C57Bl/6 mice over a 15-week period, comparing overall status and body weight change. Practical applications: This work provides a complete description of (i) an efficient and cost-effective synthesis of triheptanoin and (ii) its formulation as a solid, stable, and palatable ketogenic diet (triheptanoin-rich; 39% of the caloric intake) for rodents. Triheptanoin-rich diets will be helpful on pre-clinical experiments testing the therapeutic efficacy of triheptanoin in different rodent models of human diseases. In addition, using the same solidification procedure, other oils could be incorporated into rodent ketogenic diet to study their dosage and long-term effects on mammal health and development. This approach could be extremely valuable as ketogenic diet is widely used clinically for epilepsy treatment.

Polyphenol-rich foods in the Mediterranean diet are associated with better cognitive function in elderly subjects at high cardiovascular risk

Brain oxidative processes play a major role in age-related cognitive decline, thus consumption of antioxidant-rich foods might help preserve cognition. Our aim was to assess whether consumption of antioxidant-rich foods in the Mediterranean diet relates to cognitive function in the elderly. In asymptomatic subjects at high cardiovascular risk (n = 447; 52% women; age 5580 y) enrolled in the PREDIMED study, a primary prevention dietary-intervention trial, we assessed food intake and cardiovascular risk profile, determined apolipoprotein E genotype, and used neuropsychological tests to evaluate cognitive function.We also measured urinary polyphenols as an objective biomarker of intake. Associations between energy-adjusted food consumption, urinary polyphenols, and cognitive scores were assessed by multiple linear regression models adjusted for potential confounders. Consumption of some foods was independently related to better cognitive function. The specific associations [regression coefficients (95% confidence intervals)] were: total olive oil with immediate verbal memory [0.755 (0.1511.358)]; virgin olive oil and coffee with delayed verbal memory [0.163 (0.0100.316) and 0.294 (0.0550.534), respectively];walnuts with working memory [1.191 (0.0612.322)]; and wine with Mini-Mental State Examination scores [0.252 (0.0060.496)]. Urinary polyphenols were associated with better scores in immediate verbal memory [1.208 (0.2362.180)]. Increased consumption of antioxidant-rich foods in general and of polyphenols in particular is associated with better cognitive performance in elderly subjects at high cardiovascular risk. The results reinforce the notion that Mediterranean diet components might counteract age-related cognitive decline.

Intersecciones entre visión artificial y mirada artística. Semantic Intersections between Artificial and Artistic Vision.

En el presente artículo se ha desarrollado un sistema capaz de categorizar de forma automática la base de datos de imágenes que sirven de punto de partida para la ideación y diseño en la producción artística del escultor M. Planas. La metodología utilizada está basada en características locales. Para la construcción de un vocabulario visual se sigue un procedimiento análogo al que se utiliza en el análisis automático de textos (modelo 'Bag-of-Words'-BOW) y en el ámbito de las imágenes nos referiremos a representaciones 'Bag-of-Visual Terms' (BOV). En este enfoque se analizan las imágenes como un conjunto de regiones, describiendo solamente su apariencia e ignorando su estructura espacial. Para superar los inconvenientes de polisemia y sinonimia que lleva asociados esta metodología, se utiliza el análisis probabilístico de aspectos latentes (PLSA) que detecta aspectos subyacentes en las imágenes, patrones formales. Los resultados obtenidos son prometedores y, además de la utilidad intrínseca de la categorización automática de imágenes, este método puede proporcionar al artista un punto de vista auxiliar muy interesante.

Cocoa flavonoid-enriched diet modulates systemic and intestinal immunoglobulin synthesis in adult Lewis rats

Previous studies have reported that a diet containing 10% cocoa, a rich source of flavonoids, has immunomodulatory effects on rats and, among others effects, is able to attenuate the immunoglobulin (Ig) synthesis in both systemic and intestinal compartments. The purpose of the present study was focused on investigating whether these effects were attributed exclusively to the flavonoid content or to other compounds present in cocoa. To this end, eight-week-old Lewis rats were fed, for two weeks, either a standard diet or three isoenergetic diets containing increasing proportions of cocoa flavonoids from different sources: one with 0.2% polyphenols from conventional defatted cocoa, and two others with 0.4% and 0.8% polyphenols, respectively, from non-fermented cocoa. Diet intake and body weight were monitored and fecal samples were obtained throughout the study to determine fecal pH, IgA, bacteria proportions, and IgA-coated bacteria. Moreover, IgG and IgM concentrations in serum samples collected during the study were quantified. At the end of the dietary intervention no clear changes of serum IgG or IgM concentrations were quantified, showing few effects of cocoa polyphenol diets at the systemic level. However, in the intestine, all cocoa polyphenol-enriched diets attenuated the age-related increase of both fecal IgA and IgA-coated bacteria, as well as the proportion of bacteria in feces. As these effects were not dependent on the dose of polyphenol present in the diets, other compounds and/or the precise polyphenol composition present in cocoa raw material used for the diets could be key factors in this effect.

Modulation in Wistar rats of blood corticosterone compartmentation by sex and prior exposure to a cafeteria diet

In the metabolic syndrome, glucocorticoid activity is increased, but circulating levels show little change. Most of blood glucocorticoids are bound to corticosteroid-binding globulin (CBG), which liver expression and circulating levels are higher in females than in males. Since blood hormones are also bound to blood cells, and the size of this compartment is considerable for androgens and estrogens, we analyzed whether sex or eating a cafeteria diet altered the compartmentation of corticosterone in rat blood. The main corticosterone compartment in rat blood is that specifically bound to plasma proteins, with smaller compartments bound to blood cells or free. Cafeteria diet increased the expression of liver CBG gene, binding plasma capacity and the proportion of blood cell-bound corticosterone. There were marked sex differences in blood corticosterone compartmentation in rats, which were unrelated to testosterone. The use of a monoclonal antibody ELISA and a polyclonal Western blot for plasma CBG compared with both specific plasma binding of corticosterone and CBG gene expression suggested the existence of different forms of CBG, with varying affinities for corticosterone in males and females, since ELISA data showed higher plasma CBG for males, but binding and Western blot analyses (plus liver gene expression) and higher physiological effectiveness for females. Good cross- reactivity to the antigen for polyclonal CBG antibody suggests that in all cases we were measuring CBG.The different immunoreactivity and binding affinity may help explain the marked sex-related differences in plasma hormone binding as sex-linked different proportions of CBG forms.

Altered nitrogen balance and decreased urea excretion in male rats fed cafeteria diet are related to arginine availability

Hyperlipidic diets limit glucose oxidation and favor amino acid preservation, hampering the elimination of excess dietary nitrogen and the catabolic utilization of amino acids.We analyzed whether reduced urea excretion was a consequence of higherNO ; (nitrite,nitrate, and other derivatives) availability caused by increased nitric oxide production in metabolic syndrome. Rats fed a cafeteria diet for 30 days had a higher intake and accumulation of amino acid nitrogen and lower urea excretion.There were no differences in plasma nitrate or nitrite. NO and creatinine excretion accounted for only a small part of total nitrogen excretion. Rats fed a cafeteria diet had higher plasma levels of glutamine, serine, threonine, glycine, and ornithinewhen comparedwith controls,whereas arginine was lower. Liver carbamoyl-phosphate synthetase I activity was higher in cafeteria diet-fed rats, but arginase I was lower. The high carbamoyl-phosphate synthetase activity and ornithine levels suggest activation of the urea cycle in cafeteria diet-fed rats, but low arginine levels point to a block in the urea cycle between ornithine and arginine, thereby preventing the elimination of excess nitrogen as urea. The ultimate consequence of this paradoxical block in the urea cycle seems to be the limitation of arginine production and/or availability.

A radiochemical method for carbamoyl-phosphate synthetase I: application to rats fed a hyperproteic diet

A method for the measurement of carbamoyl-phosphate synthetase I activity in animal tissues has been developed using the livers of rats under normal and hyperproteic diets. The method is based on the incorporation of 14C-ammonium bicarbonate to carbamoyl-phosphate in the presence of ATP-Mg and N-acetyl-glutamate. The reaction is stopped by chilling, lowering the pH and adding ethanol. Excess bicarbonate is flushed out under a gentle stream of cold CO2. The only label remaining in the medium was that incorporated into carbamoyl-phosphate, since all 14C-CO2 from bicarbonate was eliminated. The method is rapid and requires only a low pressure supply of CO2 to remove the excess substrate. The reaction is linear up to 10 min using homogenate dilutions of 1:20 to 1:200 (w/v). Rat liver activity was in the range of 89±8 nkat/g. Hyperproteic diet resulted in a significant 1.4-fold increase. The design of the method allows for the processing of multiple samples at the same time, and incubation medium manipulation is unnecessary, since the plastic incubation vial and its contents are finally counted together.

Desarrollo larvario y número de estadios larvarios de "Pseudaletia unipuncta" alimentada con dos variedades de maíz y dos dietas semisintéticas

Pseudaletia ( Mythimna) unipuncta (Haworth) es una plaga defoliadora del maíz presente año tras año en la Cuenca del Ebro, que realiza en el maíz daños esporádicos pero a veces devastadores. La implantación creciente de maíz resistente a taladros ha producido, en la zona del presente estudio, la disminución casi total de los daños debidos a los taladros Sesamia nonagrioides (Lefebvre) y Ostrinia nubilalis (Hübner). Sin embargo, no se sabe aún cómo el incremento de maíz transgénico puede afectar a las poblaciones de P. unipuncta. Actualmente no hay datos sobre biología o parámetros del desarrollo de la especie en la Península Ibérica necesarios para evaluar los posibles efectos de las nuevas variedades utilizadas. Para ello es necesario primero establecer un método de cría adecuado y posteriormente iniciar los estudios sobre la biología de la misma. En el presente trabajo se planteó conocer diferentes parámetros del desarrollo larvario de poblaciones locales de P. unipuncta alimentada sobre hoja de maíz (variedades Tietar y PR33P66) y sobre dos dietas semisintéticas, una a base de alubias. Phaseolus vulgaris, y otra a base de maíz, Zea maya. Los resultados mostraron que las larvas desarrolladas en dieta a base de alubias sufrieron menor mortalidad, que las desarrolladas con planta o sobre la otra dieta, además el peso de las pupas resultantes fue mayor y presentaron menos malformaciones. En cuanto al numero de estadios larvarios, la mayoría pupó tras seis mudas larvarias, necesitando, las de dieta de maíz, estadios larvarios adicionales. Estos resultados indican que la dieta a base de alubias es la más adecuada para posteriores trabajos con esta especie.

Effects of a post-weaning cafeteria diet in young rats: metabolic syndrome, reduced activity and low anxiety-like behaviour.

Among adolescents, overweight, obesity and metabolic syndrome are rapidly increasing in recent years as a consequence of unhealthy palatable diets. Animal models of diet-induced obesity have been developed, but little is known about the behavioural patterns produced by the consumption of such diets. The aim of the present study was to determine the behavioural and biochemical effects of a cafeteria diet fed to juvenile male and female rats, as well as to evaluate the possible recovery from these effects by administering standard feeding during the last week of the study. Two groups of male and female rats were fed with either a standard chow diet (ST) or a cafeteria (CAF) diet from weaning and for 8 weeks. A third group of males (CAF withdrawal) was fed with the CAF diet for 7 weeks and the ST in the 8th week. Both males and females developed metabolic syndrome as a consequence of the CAF feeding, showing overweight, higher adiposity and liver weight, increased plasma levels of glucose, insulin and triglycerides, as well as insulin resistance, in comparison with their respective controls. The CAF diet reduced motor activity in all behavioural tests, enhanced exploration, reduced anxiety-like behaviour and increased social interaction; this last effect was more pronounced in females than in males. When compared to animals only fed with a CAF diet, CAF withdrawal increased anxiety in the open field, slightly decreased body weight, and completely recovered the liver weight, insulin sensitivity and the standard levels of glucose, insulin and triglycerides in plasma. In conclusion, a CAF diet fed to young animals for 8 weeks induced obesity and metabolic syndrome, and produced robust behavioural changes in young adult rats, whereas CAF withdrawal in the last week modestly increased anxiety, reversed the metabolic alterations and partially reduced overweight.