Pan-Genome-Wide Association Study reveals a key role of the salmochelin receptor IroN in the biofilm formation of Salmonella Typhimurium and its monophasic variant 4,[5],12:i:.

Salmonella enterica subsp. enterica serovar Typhimurium variant 4,[5],12:i:- (so called S. 4,[5],12:i:-) has rapidly become one of the most prevalent serovars in humans in Europe, with clinical cases associated with foodborne from pork products. The mechanisms, genetic basis and biofilms relevance by which S. 4,[5],12:i:- maintains and spreads its presence in pigs remain unclear. In this study, we examined the genetic basis of biofilm production in 78 strains of S. 4,[5],12:i:- (n = 57) and S. Typhimurium (n = 21), from human gastroenteritis, food products and asymptomatic pigs. The former showed a lower Specific Biofilm Formation index (SBF) and distant phylogenetic clades, suggesting that the ability to form biofilms is not a crucial adaptation for the S. 4,[5],12:i:- emerging success in pigs. However, using a pan-Genome-Wide Association Study (pan-GWAS) we identified genetic determinants of biofilm formation, revealing 167 common orthologous groups and genes associated with the SBF. The analysis of annotated sequences highlighted specific genetic deletions in three chromosomal regions of S. 4,[5],12:i:- correlating with SBF values: i) the complete fimbrial operon stbABCDE widely recognized as the most critical factor involved in Salmonella adherence; ii) the hxlA, hlxB, and pgiA genes, which expression in S. Typhimurium is induced in the tonsils during swine infection, and iii) the entire iroA locus related to the characteristic deletion of the second-phase flagellar genomic region in S. 4,[5],12:i:-. Consequently, we further investigated the role of the iro-genes on biofilm by constructing S. Typhimurium deletion mutants in iroBCDE and iroN. While iroBCDE showed no significant impact, iroN clearly contributed to S. Typhimurium biofilm formation. In conclusion, the pan-GWAS approach allowed us to uncover complex interactions between genetic and phenotypic factors influencing biofilm formation in S. 4,[5],12:i:- and S. Typhimurium.

Valorisation of cellulosic rejections from wastewater treatment plants through sugar production.

The widespread use of wipes and other sanitary products made of nonwoven fibres has led to an enormous problem in wastewater treatment systems that has been underestimated for some time. To date, there are no practical alternatives for recycling and valorisation. In this study, cellulosic rejections recovered from a wastewater treatment plant in Barcelona (Spain) were characterised and treated using hydrothermal and enzymatic methods to obtain free sugars. Steam explosion and autoclave pre-treatments were performed at different temperatures (120, 130, or 150 °C) and residence times (10-40 min) under neutral, acidic or basic conditions. The solids obtained after the pre-treatment, as well as the untreated material, were subjected to enzymatic hydrolysis using commercial enzymes. The untreated substrate reached the highest sugar production: 29 g glucose and xylose per 100 g of the cellulosic rejections, equivalent to 86% of the sugars contained in the initial material. These sugars can subsequently be transformed into biofuels or bioproducts within a biorefinery approach.

Relationship between Salmonella infection, shedding and serology in fattening pigs in low-moderate prevalence areas.

Salmonella is a major foodborne pathogen causing important zoonosis worldwide. Pigs asymptomatically infected in mesenteric lymph nodes (MLN) can be intermittent shedders of the pathogen through faeces, being considered a major source of human infections. European baseline studies of fattening pig salmonellosis are based on Salmonella detection in MLN. This work studies the relationship between Salmonella infection in MLN and intestinal content (IC) shedding at slaughter and the relationship between the presence of the pathogen and the serologic status at farm level. Mean Salmonella prevalence in the selected pigs (vertically integrated production system of Navarra, Spain) was 7.2% in MLN, 8.4% in IC and 9.6% in serum samples. In this low-moderate prevalence context, poor concordance was found between MLN infection and shedding at slaughter and between bacteriology and serology. In fact, most of shedders were found uninfected in MLN (83%) or carrying different Salmonella strains in MLN and in IC (90%). The most prevalent Salmonellae were Typhimurium resistant to ACSSuT ± Nx or ASSuT antibiotic families, more frequently found invading the MLN (70%) than in IC (33.9%). Multivariable analysis revealed that risk factors associated with the presence of Salmonella in MLN or in IC were different, mainly related either to good hygiene practices or to water and feed control, respectively. Overall, in this prevalence context, detection of Salmonella in MLN is an unreliable predictor of faecal shedding at abattoir, indicating that subclinical infections in fattening pigs MLN could have limited relevance in the IC shedding.

Sugar production from barley straw biomass pretreated by combined alkali and enzymatic extrusion.

A pretreatment that combines a thermo-mechanical process (extrusion) with chemical and biological catalysts to produce fermentable sugars from barley straw (BS) biomass was investigated. BS was firstly extruded with alkali and then, the pretreated material (extrudate) was submitted to extrusion with hydrolytic enzymes (bioextrusion). The bioextrudate was found to have 35% (w/w dwb) of total solids in soluble form, partly coming from carbohydrate hydrolysis during bioextrusion. About 48% of soluble solids dry weight is comprised by sugars, mostly glucose and xylose. Further enzymatic hydrolysis of bioextrudate could be successfully carried out at high solid loading level of 30% (w/v), with sugar production yield of 32 g glucose and 18 g xylose/100g bioextrudate at 72 h incubation (equivalent to 96 and 52 g/l concentration, respectively). These results, together with the high level of integration of the process, indicate a great potential of this pretreatment technology for sugar production from lignocellulosic substrates.

Ethanol production from glucose and xylose obtained from steam exploded water-extracted olive tree pruning using phosphoric acid as catalyst.

In this work, the effect of phosphoric acid (1% w/w) in steam explosion pretreatment of water extracted olive tree pruning at 175°C and 195°C was evaluated. The objective is to produce ethanol from all sugars (mainly glucose and xylose) contained in the pretreated material. The water insoluble fraction obtained after pretreatment was used as substrate in a simultaneous saccharification and fermentation (SSF) process by a commercial strain of Saccharomyces cerevisiae. The liquid fraction, containing mainly xylose, was detoxified by alkali and ion-exchange resin and then fermented by the xylose fermenting yeast Scheffersomyces stipitis. Ethanol yields reached in a SSF process were close to 80% when using 15% (w/w) substrate consistency and about 70% of theoretical when using prehydrolysates detoxified by ion-exchange resins. Considering sugars recovery and ethanol yields about 160g of ethanol from kg of water extracted olive tree pruning could be obtained.

Daidzein has neuroprotective effects through ligand-binding-independent PPARγ activation.

Phytoestrogens are a group of plant-derived compounds that include mainly isoflavones like daidzein. Phytoestrogens prevent neuronal damage and improve outcome in experimental stroke; however, the mechanisms of this neuroprotective action have not been fully elucidated. In this context, it has been postulated that phytoestrogens might activate the peroxisome proliferator-activated receptor-γ (PPARγ), which exerts neuroprotective effects in several settings. The aim of this study was to determine whether the phytoestrogen daidzein elicits beneficial actions in neuronal cells by mechanisms involving activation of PPARγ. Our results show that daidzein (0.05-5 µM) decreases cell death induced by exposure to oxygen-glucose deprivation (OGD) from rat cortical neurons and that improves synaptic function, in terms of increased synaptic vesicle recycling at nerve terminals, being both effects inhibited by the PPARγ antagonist T0070907 (1 µM). In addition, this phytoestrogen activated PPARγ in neuronal cultures, as shown by an increase in PPARγ transcriptional activity. Interestingly, these effects were not due to binding to the receptor ligand site, as shown by a TR-FRET PPARγ competitive binding assay. Conversely, daidzein increased PPARγ nuclear protein levels and decreased cytosolic ones, suggesting nuclear translocation. We have used the receptor antagonist (RE) fulvestrant to study the neuroprotective participation of daidzein via estrogen receptor and at least in our model, we have discarded this pathway. These results demonstrate that the phytoestrogen daidzein has cytoprotective properties in neurons, which are due to an increase in PPARγ activity not mediated by direct binding to the receptor ligand-binding domain but likely due to post-translational modifications affecting its subcellular location and not depending to the RE and it is not additive with the agonist rosiglitazone.

Utilidad del índice FRAX en la evaluación de osteoporosis en pacientes con EPOC candidatos a trasplante pulmonar / Invassive pulmonary aspergillosis with myocardial involvement

Introducción: La presencia de osteoporosis (OTP) añade comorbilidad al proceso del trasplante pulmonar (TxP). Su identificación y tratamiento disminuirá el riesgo de fracturas. Objetivos: Comparar la utilidad (..) (AU)

Introduction: The presence of osteoporosis (OTP) adds comorbidity to the lung transplant (LTx) process. Its identification and treatment (..) (AU)

Tests neurológicos para la evaluación del pronóstico funcional en modelos de ictus isquémico en roedores / Neurological tests for functional outcome assessment in rodent models of ischaemic stroke

Un aspecto crítico en todos los modelos experimentales de patologías del sistema nervioso es la evaluación del pronóstico neurológico final. En el caso de una lesión cerebral isquémica focal, además de la determinación del tamaño de la lesión, una valiosa herramienta es la evaluación del déficit funcional final. Ello se debe al hecho de que el daño isquémico produce diferentes grados de deterioro sensoriomotor y cognitivo, que pueden proporcionar información sobre la ubicación y el tamaño de la lesión y sobre la eficacia de los tratamientos neuroprotectores después del daño agudo. Además, la magnitud de estas alteraciones también puede ser útil para predecir el resultado final y para evaluar terapias reparadoras a largo plazo. Con este fin se ha desarrollado una amplia gama de tests que permite la cuantificación de todos estos síntomas neurológicos. Esta revisión tiene como intención recopilar los tests de comportamiento más útiles diseñados para evaluar los síntomas neurológicos en los estudios de isquemia cerebral focal experimental en roedores inducida por oclusión de la arteria cerebral media, el modelo más utilizado para el estudio del ictus isquémico (AU)

A critical aspect in all models is the assessment of the final outcome of the modelling procedure. In the case of a focal ischaemic brain injury, apart from the determination of the size of the lesion, another valuable tool is the evaluation of the final functional deficit. Indeed, ischaemic damage leads to the appearance of different degrees of sensoriomotor and cognitive impairments, which may yield useful information on location and size of the lesion and on the efficacy of neuroprotective treatments after the acute injury. In addition, the magnitude of these impairments may also be useful to predict final outcome and to evaluate neuro-restorative therapies in a long-term scenario. To this aim, a wide range of tests has been developed which allow the quantification of all these neurological symptoms. This review intends to compile the most useful behavioural tests designed to assess neurological symptoms in studies of focal experimental cerebral ischemia in rodents induced by middle cerebral artery occlusion, the most commonly used model of ischaemic stroke (AU)

Effect of water extraction on sugars recovery from steam exploded olive tree pruning.

Biomass of olive tree pruning can be considered a suitable raw material for the production of ethanol due to its high content of potentially fermentable carbohydrates. However its high extractives content could cause condensation reactions between extractives and acid insoluble lignin during pretreatment, hindering the enzymatic hydrolysis of pretreated material. In this work, the effect of extractives removal before steam explosion of olive tree pruning was evaluated. The objectives are to recover as much glucose as possible in the extraction stage and to avoid the condensation reactions. The effect of temperature and time of water extracted material on sugars recovery was studied using a response surface method according to a central composite design. Extractive removal previous to steam explosion resulted in 20% more total sugars recovery in comparison to a material without water extraction stage.

Application of Fenton's reaction to steam explosion prehydrolysates from poplar biomass.

The application of Fenton's reaction to enhance the fermentability of prehydrolysates obtained from steam explosion pretreatment of poplar biomass was studied. Reaction conditions of temperature and H2O2 and Fe(II) concentrations were studied. The fermentability of prehydrolysate treated by Fenton's reaction was tested by using different inoculum sizes of thermotolerant strain Kluyveromyces marxianus CECT 10875. The highest percentages of toxic compound degradation (ranging from 71 to 93% removal) were obtained at the highest H2O2 concentration tested (50 mM). However, a negative effect on fermentability was observed at this H2O2 concentration at the lower inoculum loading. An increase in inoculum size to 0.6 g/L resulted in an enhanced ethanol fermentation yield of 95% relative to control.

RYS1, a foldback transposon, is activated by tissue culture and shows preferential insertion points into the rye genome.

The study of two variable amplicons of rye indicates that RYS1, a mobile element, is activated during tissue culture. We propose that RYS1 could be a foldback (FB) transposon. The FB transposons have been rarely reported in plants; RYS1 is the first described in rye and also the first active plant FB transposon reported. Preferential integration points in the rye genome exist, because the new insertions seem to be located, in all studied cases, in the same genome positions. We assume that RYS1 became active in rye very recently, as different plants from in vivo-growing cultivars showed that these elements were present or absent in the same genomic position in which the in vitro-activated element was found. This high rate of modification in these particular loci, both in the in vivo and in vitro populations, could indicate that probably the mechanisms promoting genetic variability in nature are the same that induce variation in vitro, and the modifications induced by somaclonal variation could be already present in vivo populations.

Ethanol production from lignocellulosic byproducts of olive oil extraction.

The recent implementation of a new two-step centrifugation process for extracting olive oil in Spain has substantially reduced water consumption, thereby eliminating oil mill wastewater. However, a new high sugar content residue is still generated. In this work the two fractions present in the residue (olive pulp and fragmented stones) were assayed as substrate for ethanol production by the simultaneous saccharification and fermentation (SSF) process. Pretreatment of fragmented olive stones by sulfuric acid-catalyzed steam explosion was the most effective treatment for increasing enzymatic digestibility; however, a pretreatment step was not necessary to bioconvert the olive pulp into ethanol. The olive pulp and fragmented olive stones were tested by the SSF process using a fed-batch procedure. By adding the pulp three times at 24-h intervals, 76% of the theoretical SSF yield was obtained. Experiments with fed-batch pretreated olive stones provided SSF yields significantly lower than those obtained at standard SSF procedure. The preferred SSF conditions to obtain ethanol from olives stones (61% of theoretical yield) were 10% substrate and addition of cellulases at 15 filter paper units/g of substrate.

Effect of chip size on steam explosion pretreatment of softwood.

Although considerable progress has been made in technology for converting lignocellulosic biomass into ethanol, substantial opportunities still exist to reduce production costs. In biomass pretreatment, reducing milling power is a technological improvement that will substantially lower production costs for ethanol. Improving sugar yield from hemicellulose hydrolysis would also reduce ethanol production costs. Thus, it would be desirable to test innovative pretreatment conditions to improve the economics by reducing electrical power of the milling stage and by optimizing pretreatment recovery of hemicellulose, as well as to enhance cellulose hydrolysis. The objective of this study was to evaluate the effect of chip size (2-5, 5-8, and 8-12 mm) on steam-explosion pretreatment (190 and 210 degrees C, 4 and 8 min) of softwood (Pinus pinaster).

Effect of surfactants and zeolites on simultaneous saccharification and fermentation of steam-exploded poplar biomass to ethanol.

In this work, the effect of the addition of different concentrations of Tween-80 and three different zeolite-like products on enzymatic hydrolysis, ethanol fermentation, and simultaneous saccharification and fermentation (SSF) process has been investigated. The ability of these products to enhance the effectiveness of the SSF process to ethanol of steam-exploded poplar biomass using the thermotolerant strain Kluyveromyces marxianus EMS-26 has been tested. Tween-80 (0.4 g/L) increased enzymatic hydrolysis yield by 20% when compared to results obtained in hydrolysis in absence of the additive. Zeolite-like products (ZESEP-56 and ZECER-56) (2.5 g/L) improved rates of conversion and ethanol yields in the fermentation of liquid fraction recovered from steam-exploded poplar. The periods required for the completion of fermentation were approx 10 h in the presence of zeolite-like products and 24 h in the absence of additives. The probable mode of action is through lowered levels of inhibitory substances because of adsorption by the additive.

[Invasive Haemophilus influenzae type B infections in infancy (1981-1990)]. / Infecciones invasivas por Haemophilus influenzae tipo B en la infancia (1981-1990).

We report the epidemiology of invasive Haemophilus influenzae type B infections. The study is based on 58 children and was carried out during the years from 1981 to 1990. Sixty-three percent of the patients had meningitis, 10.3% had arthritis, 10.3% had epiglottitis, 8.6% had cellulitis and 6.9% had pneumonia. Thirteen percent of the cases were children who were less than 6 months old, 34.4% were less than 12 months old, 70.6% were less than 24 months of age and 93% were less than 4 years old. Among the latter, 90% of the children with epiglottis or pneumonia were 24 months or older compared with 16% of the patients with meningitis, arthritis or cellulitis (p < 0.001). Sixty-three percent of the isolated strains of Haemophilus influenzae type B were resistant to ampicillin and 19.2% of the strains were resistant to ampicillin and chloramphenicol.

Optimization of the simultaneous saccharification and fermentation process using thermotolerant yeasts.

Different treatments to improve the thermotolerance of fermenting yeasts for simultaneous ethanol saccharification and fermentation process of cellulosic materials have been examined. Yeasts of the genera Saccharomyces and Kluyveromyces were tested for growth and fermentation at progressively higher temperatures in the range of 42-47 degrees C. The best results were obtained with K. marxianus LG, which was then submitted to different treatments in order to achieve thermotolerant clones. A total of 35 new clones were obtained that dramatically improved the SSF of 10% Solka-floc substrate at 45 degrees C when compared to the original strain, some with ethanol concentrations as high as 33 g/L.

Selection of thermotolerant yeasts for simultaneous saccharification and fermentation (SSF) of cellulose to ethanol.

A total of 27 yeast strains belonging to the groups Candida, Saccharomyces, and Kluyveromyces were screened for their ability to grow and ferment glucose at temperatures ranging 32-45 degrees C. K. marxianus and K. fragilis were found to be the best ethanol producing organisms at the higher temperature tested and, so, were selected for subsequent simultaneous saccharification and fermentation (SSF) studies. SSF experiments were performed at 42 and 45 degrees C, utilizing Solkafloc (10%) as cellulose substrate and a cellulase loading of 15 FPU/g substrate. Best results were achieved at 42 degrees C with K. marxianus L. G. and K. fragilis L. G., both of which produced close to 38 g/L ethanol and 0.5 ethanol yield, in 78 h.